PMID- 19184525 TI - First case of Microsporum ferrugineum from Tunisia. AB - This is the first case of Microsporum ferrugineum isolated from a Tunisian patient. A 60-year-old man was admitted for tinea sycosis associated with circinate herpes of the hand. Examination disclosed diffuse erythematic and perifollicular papules and pustules in the beard area. Typical ringworm vesiculo pustular lesions involved skin of the hand. Isolates were identified as Microsporum sp on the basis of macroscopic and microscopic colony characteristics. The diagnosis of M. ferrugineum was confirmed by PCR sequencing of Chitin Synthase1 gene. The patient was treated successfully with Griseofulvin, which was administered for 4 weeks. PMID- 19184526 TI - Histoplasma capsulatum recovery from the urine and a short review of genitourinary histoplasmosis. AB - Although virtually any organ can be involved in disseminated histoplasmosis, the recovery of Histoplasma capsulatum from the urine is a rare finding. Here we describe that a renal transplant recipient had H. capsulatum recovered from urinary sediment. The organism was also recovered from urine cultures. The potential implications of this finding are discussed, and the literature on genitourinary histoplasmosis is reviewed. PMID- 19184527 TI - Does age modify the relationship between morbidity severity and physical health in English and Dutch family practice populations? AB - PURPOSE: To investigate the co-influences of age and morbidity severity on physical health in adult family practice populations. METHODS: Morbidity data in a 12-month period for 7,833 older English consulters aged 50 years and over and 6,846 Dutch consulters aged 18 years and over was linked to their physical health status obtained from cross-sectional health surveys. Individual patients were categorised using 78 consulting morbidities classified by a chronicity measure (acute, acute-on-chronic and chronic) into an ordinal scale of morbidity severity ranging from single to multiple chronicity groups. Associations between morbidity severity, age and SF-12 Physical Component Summary (PCS) score were assessed using linear regression methods. RESULTS: Increased age and higher morbidity severity were significantly associated with poorer physical health. Of the explained total variance in adjusted PCS scores, an estimated 43% was attributed to increasing age, 40% to morbidity severity and 17% to deprivation for English consulters; the figures were 21, 42 and 31%, respectively for Dutch consulters. The largest differences in PCS scores between severity categories were observed in the younger age groups. CONCLUSIONS: Morbidity severity and age mainly act separately in adversely influencing physical health. In ageing populations who will experience higher multimorbidity, this study underlines the importance that health care and public health will need to address morbidity severity and ageing as related but distinct issues. PMID- 19184528 TI - Structural analysis reveals DNA binding properties of Rv2827c, a hypothetical protein from Mycobacterium tuberculosis. AB - Tuberculosis (TB) is a major global health threat caused by Mycobacterium tuberculosis (Mtb). It is further fueled by the HIV pandemic and by increasing incidences of multidrug resistant Mtb-strains. Rv2827c, a hypothetical protein from Mtb, has been implicated in the survival of Mtb in the macrophages of the host. The three-dimensional structure of Rv2827c has been determined by the three wavelength anomalous diffraction technique using bromide-derivatized crystals and refined to a resolution of 1.93 A. The asymmetric unit of the orthorhombic crystals contains two independent protein molecules related by a non crystallographic translation. The tertiary structure of Rv2827c comprises two domains: an N-terminal domain displaying a winged helix topology and a C-terminal domain, which appears to constitute a new and unique fold. Based on structural homology considerations and additional biochemical evidence, it could be established that Rv2827c is a DNA-binding protein. Once the understanding of the structure-function relationship of Rv2827c extends to the function of Rv2827c in vivo, new clues for the rational design of novel intervention strategies may be obtained. PMID- 19184529 TI - X-ray crystal structure of GarR-tartronate semialdehyde reductase from Salmonella typhimurium. AB - Tartronate semialdehyde reductases (TSRs), also known as 2-hydroxy-3 oxopropionate reductases, catalyze the reduction of tartronate semialdehyde using NAD as cofactor in the final stage of D-glycerate biosynthesis. These enzymes belong to family of structurally and mechanically related beta-hydroxyacid dehydrogenases which differ in substrate specificity and catalyze reactions in specific metabolic pathways. Here, we present the crystal structure of GarR a TSR from Salmonella typhimurium determined by the single-wavelength anomalous diffraction method and refined to 1.65 A resolution. The active site of the enzyme contains L-tartrate which most likely mimics a position of a glycerate which is a product of the enzyme reaction. The analysis of the TSR structure shows also a putative NADPH binding site in the enzyme. PMID- 19184530 TI - A banking strategy toward customized therapy in breast cancer. AB - In breast cancer, various clinical parameters are assessed to define clinical stage and thus obtain a more accurate prognosis. However, banks of tumor tissues are an important source of material for studies of risk of recurrence and of features governing clinical outcome in breast cancer. Although the heterogeneous characteristics of individual tumors, subtle phenotypes and stem cells can only be identified in viable cells, tissue banks often give low priority to the preservation of living cells because it is labor-intensive and expensive. The present study was designed to evaluate the feasibility of introducing, within the routine procedures of tissue preservation, a cryopreservation protocol that allows the recovery of living cells after storage. We analyzed the effect of storage time on cell viability, growth rates, and protein expression of ten human breast cancer specimens subjected to various cryopreservation techniques. Cryopreservation of cancer tissue specimens for 12 months allowed protein characterization but not the recovery of living cells. Here we show that enzymatic digestion immediately before slow freezing, and storage in liquid nitrogen permits the recovery and expansion of living cells that can be tailored to specific requirements and projects. PMID- 19184531 TI - The importance of the values attached to cells for a good informed consent procedure in cell donation for tissue engineering purposes. AB - In this article the scope and contents of informed consent for cell donation in tissue engineering are discussed. We evaluate the various values donors may attach to cells and that may influence their decision to donate. Awareness of the various meanings of the value of the cells is therefore essential for all involved in cell transfer. We discuss four types of value: biological, relational, material and informational value and discuss their relevance for collection, for the relationships that are established between donor and cell collector and for informed consent procedures. The actual setting of cell donation and the contents of informed consent can either maintain or disrupt these relationships. While one can exclude donors from any further say in the destination of their cells and from any financial return, one cannot exclude them from a persistent interest in the genetic information of the cells. PMID- 19184532 TI - Comparison of two cellular harvesting methods for primary human oral culture of keratinocytes. AB - The possibility of obtaining transplantable oral epithelia opens new perspectives for oral treatments. Most of them are surgical, resulting in mucosal failures. As reconstructive material this in vitro epithelia would be also useful for other parts of the human body. Many researchers still use controversial methods; therefore it was evaluated and compared the efficiency of the enzymatic and direct explant methods to obtain oral keratinocytes. To this project oral epithelia fragments were used. This work compared: time needed for cell obtainment, best cell amount, life-span and epithelia forming cell capacity. The results showed the possibility to obtain keratinocytes from a small oral fragment and we could verify the advantages and peculiar restrictions. We concluded that under our conditions the enzymatic method showed the best results: in the cells obtaining time needed, cell amount and life-span. Both methods showed the same capacity to form in vitro epithelia. PMID- 19184533 TI - Brain donation: who and why? AB - Understanding what influences people to donate, or not donate, body organs and tissues is very important for the future of transplant surgery and medical research (Garrick in J Clin Neurosci 13:524-528, 2006). A previous web-based motivation survey coordinated by the New South Wales Tissue Resource Centre found that most people who participated in brain donation were young, female, educated Australians, not affiliated with any particular religion, and with a higher prevalence of medical illness than the general Australian population. It discussed the main motivating factors for brain donation to be "the benefits of the research to medicine and science". This study has been replicated in a paper based version to capture a broader cross-section of the general population, to find out who they are and what motivates them to donate. All consented and registered brain donors (n = 1,323) were sent a questionnaire via the post and recipients were given 3 months to complete the questionnaire and return it in a reply paid envelope. Results were entered into the original web-based survey and analyzed using SPSS version 10. Six hundred and fifty-eight questionnaires were returned completed, a response rate of 53%. The results show that people from all age groups are interested in brain donation. The over 65's are the largest of the groups (30.7%). The majority of the participants were female (60.6%), married (49.2%) with children (65.8%), employed (52.9%) and have a tertiary education (73.3%). They were either non-religious (48.2%) or Christian (41.6%) and were mostly Australian (65.4%). Most (81%) had pledged to donate other organs and tissues for transplantation. The most commonly cited reasons for the donation were to benefit science (27.6%), to benefit medicine (23.9%), a family illness (17.5%) and to benefit the community (16.6%). This study demonstrates that people across all age groups are interested in brain donation. Recruitment of new brain donors could target the over 65 female Australians, who are not religious or Christian and who have also donated other organs and tissues for transplant purposes. It also indicates the need to make donation for research part of the national transplant donation program. PMID- 19184534 TI - Health behaviors among Ashkenazi Jewish individuals receiving counseling for BRCA1 and BRCA2 mutations. AB - Little of the research involving individuals undergoing testing for BRCA1/2 mutations has examined the effect of genetic counseling and testing on diet, exercise, and vitamin use. This study examined diet, exercise, and vitamin use among 120 Ashkenazi Jewish individuals with a personal and/or family history of breast and/or ovarian cancer who presented for BRCA1/2 genetic counseling and testing. Health behaviors were measured at pre-counseling and 6 months post results. T-tests or ANOVAs were used to test for association of health behaviors with potential predictors: personal cancer history, anxiety, perceived cancer risk, gene status, and perceived efficacy of health behaviors. Multivariable linear regression analysis found gender (P = .007) and perceived efficacy of a healthy diet (P = .005) to be significantly associated with healthy food consumption; gender (P = .003), education (P = .01), and personal cancer history (P = .02) were significantly associated with unhealthy food consumption; higher age (P < .001) and perceived efficacy (P < .001) were significantly associated with higher vitamin use. Paired t-tests found no significant changes in diet, vitamin use, and exercise as a result of genetic counseling and testing. We conclude that genetic counseling and testing had little impact on health behaviors. Rather, perceived efficacies of certain health behaviors to prevent or delay cancer may be more reliable predictors of those behaviors. Clinicians should assess perceived efficacies of health behaviors in their interventions to improve healthy practices in the BRCA1/2 genetic testing population. PMID- 19184535 TI - SDHB-associated renal oncocytoma suggests a broadening of the renal phenotype in hereditary paragangliomatosis. AB - Mutations in SDHB are one of the causes of hereditary paraganglioma syndrome. Germline mutations in SDHB predispose to the development of head and neck paragangliomas and phaeochromocytomas. Renal tumours are also increasingly being reported as component tumours in hereditary paragangliomatosis associated with mutations in SDHB. We present the first reported case of a family in whom an individual shown to carry a mutation in SDHB developed a renal oncocytoma. We review other reports of renal tumours associated with SDHB-associated hereditary paragangliomatosis and suggest that various histological subtypes of renal tumours are part of this condition. This observation indicates that SDHB associated hereditary paragangliomatosis is unlike most tumour predisposition syndromes associated with the development of renal tumours which are usually associated with specific histological sub-types. The increasing recognition of the involvement of renal tumours in SDHB mutation carriers suggests that renal screening is likely to be valuable for these patients. SDHB mutations should also be considered in the context of genetic testing when renal tumours, regardless of histopathology, present in families with other tumours consistent hereditary paraganglioma syndrome. PMID- 19184536 TI - Recent improvements in the development of A(2B) adenosine receptor agonists. AB - Adenosine is known to exert most of its physiological functions by acting as local modulator at four receptor subtypes named A(1), A(2A), A(2B) and A(3) (ARs). Principally as a result of the difficulty in identifying potent and selective agonists, the A(2B) AR is the least extensively characterised of the adenosine receptors family. Despite these limitations, growing understanding of the physiological meaning of this target indicates promising therapeutic perspectives for specific ligands. As A(2B) AR signalling seems to be associated with pre/postconditioning cardioprotective and anti-inflammatory mechanisms, selective agonists may represent a new therapeutic group for patients suffering from coronary artery disease. Herein we present an overview of the recent advancements in identifying potent and selective A(2B) AR agonists reported in scientific and patent literature. These compounds can be classified into adenosine-like and nonadenosine ligands. Nucleoside-based agonists are the result of modifying adenosine by substitution at the N (6)-, C(2)-positions of the purine heterocycle and/or at the 5'-position of the ribose moiety or combinations of these substitutions. Compounds 1-deoxy-1-{6-[N'-(furan-2-carbonyl)-hydrazino] 9H-purin-9-yl}-N-ethyl-beta-D-ribofuranuronamide (19, hA(1) K (i) = 1050 nM, hA(2A) K (i) = 1550 nM, hA(2B) EC(50) = 82 nM, hA(3) K (i) > 5 muM) and its 2 chloro analogue 23 (hA(1) K (i) = 3500 nM, hA(2A) K (i) = 4950 nM, hA(2B) EC(50) = 210 nM, hA(3) K (i) > 5 muM) were confirmed to be potent and selective full agonists in a cyclic adenosine monophosphate (cAMP) functional assay in Chinese hamster ovary (CHO) cells expressing hA(2B) AR. Nonribose ligands are represented by conveniently substituted dicarbonitrilepyridines, among which 2-[6-amino-3,5 dicyano-4-[4-(cyclopropylmethoxy)phenyl]pyridin-2-ylsulfanyl]acetamide (BAY-60 6583, hA(1), hA(2A), hA(3) EC(50) > 10 muM; hA(2B) EC(50) = 3 nM) is currently under preclinical-phase investigation for treating coronary artery disorders and atherosclerosis. PMID- 19184537 TI - The platelet as an immune cell-CD40 ligand and transfusion immunomodulation. AB - The discovery that platelets possess cell membrane, cytoplasmic, and secreted forms of the co-stimulatory molecule CD40 ligand (CD40L, also known as CD154) has led to a revolution in the view of this anucleate, differentiated cell fragment, previously thought only to be involved in blood clotting (hemostasis). During the last decade, it has become clear that platelets function in innate and adaptive immunity and possess pro-inflammatory, as well as pro-thrombotic properties. They interact not only with other platelets and endothelial cells, but also with lymphocytes, dendritic cells, and structural cells such as fibroblasts. Soluble forms of CD40L (sCD40L) in the human circulation are almost entirely derived from platelets. Elevated levels of CD40L are associated with clinically important conditions, such as vascular disease, abnormal clotting (thrombosis), lung injury, and autoimmune disease. Each year millions of platelet transfusions are given to patients that contain large amounts of sCD40L. sCD40L in the supernatant of stored platelets can induce cytokines, chemokines, and lipid mediators by activating CD40 bearing cells. Increased levels of sCD40L in transfused blood are associated with transfusion-related acute lung injury, a potentially fatal complication, as well as more common, milder transfusion reactions such as fever and rigors. These effects come under the rubric of transfusion immunomodulation, which postulates that transfusion recipient biology, particularly immune function, is dramatically altered by transfusion of stored allogeneic blood. PMID- 19184538 TI - Lysophosphatidic acid and autotaxin: emerging roles in innate and adaptive immunity. AB - Lysophosphatidic acid (LPA) can affect the growth, migration, and activation of many different cell types. Research in this field has recently accelerated due to the molecular cloning of LPA receptors as well as advances in our understanding of LPA metabolism. A major pathway for LPA generation is the hydrolysis of lysophosphatidylcholine by the enzyme autotaxin (ATX). Although most research to date has been conducted in other disciplines (e.g., neurobiology and cardiovascular diseases), emerging data point to an important role for LPA and ATX in regulating immune responses. Here we review current understanding of LPA and ATX in immunity with an emphasis on migration and activation of lymphocytes and dendritic cells. New gene-targeted and transgenic mice, receptor-specific antibodies, and pathway antagonists should rapidly enhance our understanding of this versatile lysolipid in immune responses in the near future. PMID- 19184540 TI - The emerging field of osteoimmunology. AB - Recent studies have elucidated unanticipated connections between the immune and skeletal systems, and this relationship has led to the development of a new field known as osteoimmunology. The goal of research in this field is to: (1) further understand how the bone microenvironment influences immune cell ontogeny and subsequent effector functions, and (2) translate basic science findings in bone biology to clinical applications for autoimmune diseases that target the skeleton such as rheumatoid arthritis (RA). In this review, we will examine the recent findings of the interplay between the immune and skeletal systems. This discussion will focus on the cells and signaling pathways in osteoimmune interactions and how innate and adaptive immune effector cells as well as cytokines and chemokines play a role in the maintenance and dysregulation of skeletal-immune homeostasis. We will also discuss how immunomodulatory biologic drugs, which specifically target these cells and effector molecules, have transformed the treatment of autoimmune mediated inflammatory diseases (IMIDs) and metabolic bone diseases such as osteoporosis. PMID- 19184539 TI - Leukocyte integrins and their ligand interactions. AB - Although critical for cell adhesion and migration during normal immune-mediated reactions, leukocyte integrins are also involved in the pathogenesis of diverse clinical conditions including autoimmune diseases and chronic inflammation. Leukocyte integrins therefore have been targets for anti-adhesive therapies to treat the inflammatory disorders. Recently, the therapeutic potential of integrin antagonists has been demonstrated in psoriasis and multiple sclerosis. However, current therapeutics broadly affect integrin functions and, thus, yield unfavorable side effects. This review discusses the major leukocyte integrins and the anti-adhesion strategies for treating immune diseases. PMID- 19184541 TI - Limitations of the colloidal silica method in mapping the endothelial plasma membrane proteome of the mouse heart. AB - The endothelial cell (EC) membrane is an important interface, which plays a crucial role in signal transduction. Our aim was to selectively purify luminal EC membrane proteins from the coronary vasculature of the isolated perfused mouse heart and analyze its composition with mass spectrometry (MS). To specifically label coronary ECs in the intact heart, the colloidal silica method was applied, which is based on the binding of positively charged colloidal silica to the surface of EC membranes. Transmission electron microscopy revealed the specific labeling of ECs of macro and microvessels. Two different methods of tissue homogenization (Teflon pestle and ultra blade) together with density centrifugation were used for membrane protein enrichment. Enrichment and purity was controlled by Western blot analysis using the EC-specific protein caveolin 1 and various intracellular marker proteins. The ultra blade method resulted in a tenfold enrichment of caveolin 1, while there was negligible contamination as judged by Western blot. However, protein yield was low and required pooling of ten hearts for MS. When enriched endothelial membrane proteins were digested with trypsin and analyzed by LC-MS, a total of 56 proteins could be identified, of which only 12 were membrane proteins. We conclude that coronary endothelial membranes can be conveniently labeled with colloidal silica. However, due to the ionic nature of interaction of colloidal silica with the EC membrane the shear rate required for cardiac homogenization resulted in a substantial loss of specificity. PMID- 19184542 TI - A review of the antioxidant mechanisms of polyphenol compounds related to iron binding. AB - In this review, primary attention is given to the antioxidant (and prooxidant) activity of polyphenols arising from their interactions with iron both in vitro and in vivo. In addition, an overview of oxidative stress and the Fenton reaction is provided, as well as a discussion of the chemistry of iron binding by catecholate, gallate, and semiquinone ligands along with their stability constants, UV-vis spectra, stoichiometries in solution as a function of pH, rates of iron oxidation by O(2) upon polyphenol binding, and the published crystal structures for iron-polyphenol complexes. Radical scavenging mechanisms of polyphenols unrelated to iron binding, their interactions with copper, and the prooxidant activity of iron-polyphenol complexes are briefly discussed. PMID- 19184543 TI - Expression, purification, and characterization of hepatitis B virus surface antigens (HBsAg) in yeast Pichia Pastoris. AB - Prevention of the prevalence of HB depends upon the development of efficient diagnostic reagent and preventive vaccine. Pichia pastoris offers many advantages over the other expression systems in the production of recombinant HBsAg. In this study, we reported that the recombinant P. pastoris strains were cultured in shake flasks and then scaled up in a 5.0-l bioreactor: approximately 27 mg/l of the protein and the maximal cell OD at 600 nm of 310 were achieved in the bioreactor. The recombinant HBsAg was purified by three steps of purification procedures. SDS-PAGE showed that the purified recombinant HBsAg constituted only one homogeneous band of approximately 24 kDa. CsCl density gradient ultracentrifugation assay indicated that the density of the HBsAg was 1.2 mg/ml, which was in agreement with the natural HBsAg, the HBsAg expressed in Saccharomyces cerevisiae and in mammalian cells. Electron microscope observation revealed that the purified recombinant HBsAg was homogeneous 22-nm particles, suggesting the HBsAg expressed in P. pastoris was self-assembled to virus-like structures. Competitive ELISA indicated that P. pastoris-derived HBsAg possessed the excellent immunoreaction with anti-HBsAg. Animal immunization showed that the immunogenicity of P. pastoris-derived HBsAg was superior to that of S. cerevisiae derived HBsAg. Together, our results demonstrated that the recombinant HBsAg expressed in P. pastoris could provide promising, inexpensive, and large-scale materials for the diagnostic reagent and vaccine to prevent HBV infection. PMID- 19184544 TI - Inhibition performance of lignocellulose degradation products on industrial cellulase enzymes during cellulose hydrolysis. AB - This study examined the inhibition performance by the major lignocellulose degradation products, including organic acids, furan derivatives, lignin derivatives, and ethanol, on a broadly used commercial cellulase enzyme Spezyme CP (Genencor International, Rochester, NY, USA) to cellulose hydrolysis at both the well-mixing state (shaking flask) and the static state (test tube). The cellulase activity in the cellulase complex of Spezyme CP was assumed to be one single "cellulase", and the apparent kinetic parameters of this cellulase enzyme were measured as an approximate index of the inhibitory effect to the industrial cellulase enzyme. The inhibition performance of these degradation products was compared and analyzed using the determined apparent kinetic parameters. All the degradation products strongly inhibit the cellulose hydrolysis by cellulase enzyme, and the inhibitions on cellulase were all competitive type. The order of the inhibition strength by the lignocellulose degradation products to cellulase is lignin derivatives > furan derivatives > organic acids > ethanol. This study gave a quantitative view to the enzymatic hydrolysis of lignocellulose under the inhibition performance of the lignocellulose degradation products and will help to understand the lignocellulose recalcitrance to enzyme hydrolysis. PMID- 19184545 TI - Pretreatment and fractionation of corn stover by soaking in ethanol and aqueous ammonia. AB - A new process for pretreatment of lignocellulosic biomass, designated the soaking in ethanol and aqueous ammonia (SEAA) process, was developed to improve hemicellulose preservation in solid form. In the SEAA process, an aqueous ammonia solution containing ethanol is used. Corn stover was treated with 15 wt.% ammonia at 1:9 solid-liquid ratio (by weight) at 60 degrees C for 24 h with ethanol added at 1, 5, 20, and 49 wt.% (balance was water). The extents by which xylan was solubilized with no ethanol and with ethanol added at 1, 5, 20, and 49 wt.% of the total liquid were 17.2%, 16.7%, 14.5%, 10.4%, and 6.3% of the original xylan, respectively. Thus, at the highest ethanol concentration used the loss of hemicellulose to the liquid phase was reduced by 63%. The digestibility of glucan and xylan in the pretreated corn stover samples by cellulase was not affected by ethanol addition of up to 20 wt.%. The enzymatic digestibility of the corn stover treated with 49 wt.% ethanol added was lower than the digestibility of the sample treated with no ethanol addition. Thus, based on these results, 20 wt.% was found to be the optimum ethanol concentration for use in the SEAA process for pretreatment of corn stover. PMID- 19184546 TI - Screening and selection of marine isolate for L-glutaminase production and media optimization using response surface methodology. AB - The current work details the screening of about 400 marine isolates from various marine niches, from which one isolate was finally selected based on the productivity of glutaminase (71.23 U/l). Further, biochemical identification tests and 16S rRNA sequencing identified this isolate to be Providencia sp. This isolate was taken up for further media optimization studies by using one-factor at-a-time approach and subsequently by response surface methodology. A face centered central composite design was employed to investigate the interactive effects of four variables, viz., concentrations of glucose, methionine, urea, and succinic acid on glutaminase production. A significant influence of urea on glutaminase production was noted. Response surface methodology showed that a medium containing (g/l) glucose 10.0, urea 5.15, methionine 3.5, succinic acid 6.0, ammonium sulfate 2.5, and yeast extract 6.0 to be optimum for the production of glutaminase. The applied methodology was validated using this optimized media and enzyme activity 119 +/- 0.12 U/l and specific activity of 0.63 U/mg protein after 28 h of incubation at 25 degrees C was obtained. PMID- 19184547 TI - Isolation and characterization of low molecular weight glycosaminoglycans from marine mollusc Amussium pleuronectus (linne) using chromatography. AB - The glycosaminoglycan (GAG) heparin is a polyanionic sulfated polysaccharide most recognized for its anticoagulant activity. In the present study, the GAGs were extracted from bivalve mollusc Amussium pleuronectus. The crude GAGs were fractionated by ion-exchange (DEAE-cellulose and Amberlite IRA-900 & 120) chromatography. The recovered active fractions (as determined by metachromatic assay) were confirmed by agarose gel electrophoresis and the active fractions were purified in Sephadex G-100 column. Fractionated and purified GAG molecular weight was determined through gradient polyacrylamide gel electrophoresis. The structural characterization of low molecular weight GAG was analyzed by Fourier transform infrared spectroscopy. The activated partial thromboplastin time of purified GAG is 95 IU/mg and has molecular weight 6,500-7,500 Da. The disaccharide compositional analysis on the GAG sample was sulfated like porcine intestinal mucosal heparan sulfate, and it contains equivalent amount of uronic acid and hexosamine. The results of this study suggest that the GAG from A. pleuronectus could be an alternative source of heparin. PMID- 19184548 TI - Periodic fermentor yield and enhanced product enrichment from autonomous oscillations. AB - Four decades of work have clearly established the existence of autonomous oscillations in budding yeast culture across a range of operational parameters and in a few strains. Autonomous oscillations impact substrate conversion to biomass and products. Relatively little work has been done to quantify yield in this case. We have analyzed the yield of autonomously oscillating systems, grown under different conditions, and demonstrate that it too oscillates. Using experimental data and mathematical models of yeast growth and division, we demonstrate strategies to increase the efficient recovery of products. The analysis makes advantage of the population structure and synchrony of the system and our ability to target production within the cell cycle. While oscillatory phenomena in culture have generally been regarded with trepidation in the engineering art of bioprocess control, our results provide further evidence that autonomously oscillating systems can be a powerful tool, rather than an obstruction. PMID- 19184549 TI - Carboxylate platform: the MixAlco process part 2: process economics. AB - The MixAlco process employs a mixed culture of acid-forming microorganisms to convert biomass to carboxylate salts, which are concentrated via vapor compression evaporation and subsequently chemically converted to other chemical and fuel products. To make alcohols, hydrogen is required, which can be supplied from a number of processes, including gasifying biomass, separation from fermentor gases, methane reforming, or electrolysis. Using zeolite catalysts, the alcohols can be oligomerized into hydrocarbons, such as gasoline. A 40-tonne/h plant processing municipal solid waste ($45/tonne tipping fee) and using hydrogen from a pipeline or refinery ($2.00/kg H(2)) can sell alcohols for $1.13/gal or gasoline for $1.75/gal with a 15% return on investment ($0.61/gal of alcohol or $0.99/gal of gasoline for cash costs only). The capital cost is $1.95/annual gallon of mixed alcohols. An 800-tonne/h plant processing high-yield biomass ($60/tonne) and gasifying fermentation residues and waste biomass to hydrogen ($1.42/kg H(2)) can sell alcohols for $1.33/gal or gasoline for $2.04/gal with a 15% return on investment ($1.08/gal of alcohol or $1.68/gal of gasoline for cash costs only). The capital cost for the alcohol and gasification plants at 800 tonne/h is $1.45/annual gallon of mixed alcohols. PMID- 19184550 TI - Distribution and molecular evolution of the neuropeptide pituitary adenylate cyclase-activating polypeptide (PACAP) and its receptors in the lizard Podarcis sicula (Squamata, Lacertidae). AB - The presence of the pituitary adenylate cyclase-activating polypeptide (PACAP) and its receptors PAC(1), VPAC(1), and VPAC(2) was studied in the lizard Podarcis sicula gastrointestinal and respiratory tissues. The expression and distribution of this neuropeptide was investigated using RT-PCR, immunohistochemistry, and in situ hybridization techniques. RT-PCR showed that several tissues of this reptile synthesize an mRNA encoding for PACAP. Performing in situ hybridization and immunohistochemistry, we found a wide distribution of PACAP and its mRNA in intestine, stomach, liver, and lung. PACAP receptors possess a specific distribution in both gastrointestinal and respiratory system. Further, we analyzed the conservation of PACAP amino acid sequence demonstrating that this peptide in the lizard is very similar to that of other vertebrates. Our findings suggest that also in reptiles an effective PACAP system is present and that it could be implicated in some essential physiological functions as a result of its high conservation amongst vertebrates. PMID- 19184551 TI - Reactivity of brain tissue oxygen to change in cerebral perfusion pressure in head injured patients. AB - OBJECTIVE: It has been reported recently that correlation between brain tissue oxygen (PbtO2) and cerebral perfusion pressure (CPP) may serve as an indicator of cerebral autoregulation after subarachnoid hemorrhage. We aimed to compare similar indices describing interaction between changes in intracranial pressure (ICP), arterial blood pressure (ABP), and brain tissue oxygen to verify their clinical utility in patients after traumatic brain injury. MATERIALS AND METHODS: Retrospective analysis of multimodal monitoring of 32 patients suffering from head injury, admitted in the Neurosciences Critical Care Unit, Addenbrooke's Hospital, Cambridge, UK. Initial 24 h intervals of continuous ABP, ICP, and PbtO2 recordings were analyzed. Index of tissue oxygen reactivity ORx was evaluated as the correlation coefficient between PbtO2 and CPP over a period of 60 min and compared to the index of pressure reactivity PRx. "Optimal CPP" and a hypothetical "optimal PbtO2" were defined as the ranges of CPP and PbtO2 at which PRx or ORx were indicating best cerebrovascular milieu. RESULTS: PRx and ORx mean values did not show any correlation with each other (R = 0.012; P = 0.95) between patients. There was also no correlation between ORx and PbtO2 (R = 0.098; P = 0.61) and between PRx and PbtO2 (R = 0.019; P = 0.923). No clear and consistent value of "optimal CPP" minimizing ORx or of hypothetical "optimal PbtO2" were found analyzing PbtO2 or ORx trend over the 24 h of monitoring. However, in most of patients 'optimal CPP' has been found for PRx index. The same has been confirmed when the data from whole monitoring period were analyzed. There was no correlation between values of 'optimal CPP' assessed using ORx and 'optimal CPP' assessed with PRx. CONCLUSION: The relationships between PbtO2, ORx, and CPP in head injury appear less useful than reported before for patients after subarachnoid hemorrhage. PMID- 19184552 TI - Occurrence of perimesencephalic subarachnoid hemorrhage during pregnancy. AB - BACKGROUND: Perimesencephalic subarachnoid hemorrhage (P-SAH) is a benign subset of subarachnoid hemorrhage with a favorable prognosis and low rate of re bleeding. Risk factors may include hypertension and tobacco use, but it has not previously been reported during pregnancy. METHODS: We report two cases of P-SAH in pregnant women, a 40-year-old female, 8-weeks pregnant and a 37-year-old female at 35 weeks gestational age. RESULTS: CT scan confirmed P-SAH in both cases. CT angiography in one case and cerebral angiogram in the other did not reveal aneurysm or other potential bleeding source. The patients underwent transcranial Doppler ultrasound monitoring without evidence of vasospasm. CONCLUSION: P-SAH hemorrhage may occur during early or late pregnancy. We do not propose an increased risk of P-SAH during pregnancy. The clinical course appears favorable and CT angiography alone may be considered the preferred diagnostic test to assess for aneurysm in first trimester pregnancy. PMID- 19184554 TI - A second chance. PMID- 19184553 TI - Ventricular arrhythmia risk after subarachnoid hemorrhage. AB - INTRODUCTION: Cardiac morbidity and mortality after aneurysmal subarachnoid hemorrhage (SAH) are attributable to myocardial injury, decreased ventricular function, and ventricular arrhythmia (VA). Our objective was to test the relationships between QTc prolongation, VA, and survival after SAH. METHODS: In 200 subjects with acute aneurysmal SAH, electrocardiograms, echocardiograms, and telemetry were evaluated. Serum electrolytes and troponin were also evaluated. RESULTS: Initial QTc (mean 460 +/- 45 ms) was prolonged (> or = 470 ms) in 38% of subjects and decreased on follow-up (469 +/- 49 initial vs. 435 +/- 31 ms follow up; N = 89; P < 0.0001). VA was present in 14% of subjects, 52% of subjects with VA had QTc > or = 470 ms, and initial QTc trended toward longer duration in subjects with VA (474 +/- 61 vs. 457 +/- 42 ms; P = 0.084). Multivariate analysis demonstrated significant predictors of VA after SAH were increasing age (OR 1.3/5 years; P = 0.025), increasing stroke severity (OR 1.8; P = 0.009), decreasing heart rate (OR 0.5/10 beats/min; P = 0.006), and the absence of angiotensin converting enzyme inhibitor or angiotensin II receptor antagonist use at SAH onset (OR 0.10; P = 0.027). All-cause mortality was 19% (25/135) at 3 months and subjects with VA had significantly higher mortality than those without VA (37% vs. 16%; P = 0.027). CONCLUSIONS: These data demonstrate that QTc prolongation and arrhythmias are frequently noted after SAH, but arrhythmias are often not associated with QTc prolongation. In addition, the presence of VA identified subjects at greater risk of mortality following their SAH. PMID- 19184555 TI - Safety and efficacy of levetiracetam for critically ill patients with seizures. AB - INTRODUCTION: In intensive care unit (ICU) patients, seizure or status epilepticus treatment with intravenous benzodiazepines or conventional antiepileptic drugs (AEDs), such as phenytoin, may be accompanied by cardiovascular depression or hypotension. Levetiracetam (LVM) is a novel AED that does not undergo extensive liver metabolism, does not require drug level monitoring, and is not associated with hemodynamic instability. We retrospectively analyzed the use, safety, and efficacy of LVM in ICU patients. METHODS: Collected data included age, sex, therapy indication and duration, dosing regimen, documented seizure activity, ICU admission diagnoses, length of ICU stay, serum creatinine, liver function tests, adverse reactions, concomitant use of other AEDs, and drug interactions. RESULTS: Fifty-one patients were identified (26 males; mean (SD) age, 58.2 (19.8) years). Most patients (65%) did not receive a loading dose; the most common loading dose was 1,500 mg (50% of 18 patients). The most common maintenance dose was 500 mg twice daily (59% of 51 patients), and average duration of therapy was 13.6 (12.7) days. Approximately 47% of patients had preexisting liver disease, and 25% had elevated serum creatinine. Twenty-two patients received LVM therapy for seizure prophylaxis; 29 for acute seizure treatment. Ninety-three percent of patients treated with LVM for acute seizure had no subsequent seizures; the remaining patients (7%) required additional AEDs. One patient receiving LVM for seizure prophylaxis had documented seizures requiring additional AEDs. No adverse hemodynamic events or cardiac arrhythmias were reported. CONCLUSION: LVM appears to be safe for ICU patients when dosing is adjusted for renal function. PMID- 19184557 TI - Conventional weaning parameters do not predict extubation failure in neurocritical care patients. AB - OBJECTIVE: Predicting extubation failure (EF) is one of the most challenging aspects of critical care medicine. The literature on EF in neurocritical care patients is very scarce. We sought to determine the ability of traditional weaning parameters to predict EF in neurocritical patients. METHODS: This is a retrospective observational study performed at a tertiary level, academic, Neurological Intensive Care Unit (NCCU). We included all adult patients intubated for neurological reasons in whom an attempt to perform extubation was performed. We compared traditional weaning parameters between patients who failed extubation and those successfully extubated. Fishers exact test was used for categorical variables and t-test for continuous variables. Weaning parameters were analyzed as categorical variables and when appropriate as continuous. We incorporated a coma scale (Four Score) in an attempt to determine if neurologic dysfunction could account for EF. RESULTS: The study sample compromised 62 patients undergoing extubation trial. None of the individual weaning parameters predicted EF: rapid shallow breathing index (P = 0.62), minute ventilation (P = 0.7479), respiratory rate (P = 1.0), negative inspiratory force (P = 0.62), tidal volume, and PaO2/FIO2 ratio (P = 1.0). There was no significant difference in Four Scale score between EF and successfully extubated patients (0.44, proportions of the mean, t-test). There was no combination of weaning parameters that allowed prediction of EF. All patients had at least three normal weaning parameters, but there was no combination of parameters that accurately predicted EF. Overall, weaning parameters had better specificity than sensitivity for predicting EF. CONCLUSIONS: In this sample of neurocritical care patients undergoing extubation trial, traditional weaning parameters do not predict extubation failure. PMID- 19184556 TI - A randomized evaluation of bispectral index-augmented sedation assessment in neurological patients. AB - OBJECTIVE: To assess whether monitoring sedation status using bispectral index (BIS) as an adjunct to clinical evaluation was associated with a reduction in the total amount of sedative drug used in a 12 h period. DESIGN: Prospective randomized controlled clinical trial. SETTING: Tertiary care neurocritical care unit. PATIENTS: Sixty-seven mechanically ventilated adult patients receiving continuous intravenous sedation with propofol. INTERVENTIONS: Sedation monitoring using clinical assessment with the Ramsay scale (Ramsay-alone group) or clinical assessment plus BIS monitoring (BIS-augmentation group). Subjects were randomized to Ramsay-alone (n = 35), or BIS-augmentation (n = 32). Nurses adjusted the dose of propofol to a Ramsay of 4, or a Ramsay of 4 and BIS between 60 and 70. MEASUREMENTS AND MAIN RESULTS: Patients in the BIS-augmentation group received significantly less propofol by volume (93.5 ml vs. 157.8 ml, respectively; P < .015), and had lower infusion rates (14.6 vs. 27.9 mcg/kg/min; P = .003). There is a lower risk of propofol infusion exceeding manufacturer's recommended dosing guides in the BIS-augmentation group versus the Ramsay-alone group (0 vs. 23%, P = .0052). The BIS-augmentation group woke up much quicker than those in the Ramsay-alone group (1.2 vs. 7.5 min; P < .0001). CONCLUSIONS: BIS-augmented sedation monitoring resulted in a marked reduction in the total dose of sedative used to achieve the same level of clinical sedation resulting in shortened time to wake up without any measurable adverse effects. Physiologic sedation assessment tools may provide a useful means of improving the care of sedated critically ill patients. PMID- 19184558 TI - An analysis of potential surrogate markers of target-specific therapy in archival materials of adrenocortical carcinoma. AB - Adrenocortical carcinoma (ACC) is a rare neoplasm but some of the cases are highly malignant. Clinical outcome of the patients with advanced ACC still remained poor or dismal despite recent development of aggressive antitumor therapies. Target-specific therapies have been developed in a number of human malignancies and resulted in therapeutic benefits in some cancer patients. However, these therapies are only effective in the cases in which corresponding targets are expressed in tumor tissues. Therefore, we evaluated expression of potential surrogate markers using immunohistochemistry in archival materials of adrenocortical carcinoma in order to explore the potential application of target specific therapies in ACC in this study. We immunolocalized ten established or potential surrogate markers of target-specific therapies, located in the Ras/extracellular signal-regulated kinase and phosphatidylinositol-3 kinase/Akt pathways, in 41 ACC cases, 54 adrenocortical adenoma (ACA) cases, and five nonpathological adrenal glands and correlated the findings with clinicopathological factors of the patients. Among these markers examined, only epidermal growth factor receptor (EGFR) was significantly more abundant in ACC than in ACA (P < 0.01). These findings suggest that the agents which specifically inhibit signal transductions through EGFR such as monoclonal antibodies against EGFR are considered to be worthwhile to be attempted in future clinical studies. PMID- 19184559 TI - B-cell and T-cell epitopes in anti-factor VIII immune responses. AB - Adequate hemostasis is achieved for many hemophilia A patients by infusion of plasma-derived or recombinant factor VIII (FVIII), but unfortunately, a significant subset of patients develop an immune response in which anti-FVIII antibodies, referred to clinically as "inhibitors," interfere with its procoagulant activity. Inhibitors are the subset of anti-FVIII antibodies that bind to surfaces on FVIII (B-cell epitopes) that are important for its proper functioning in coagulation. Less antigenic FVIII molecules may be designed by identifying and then modifying the amino acid sequences of inhibitor B-cell epitopes. Conversely, characterization of these epitopes can yield important information regarding functionally important surfaces on FVIII. The production of inhibitor antibodies is driven by T cells. T cells recognize FVIII as foreign when FVIII-derived peptides bind to major histocompatibility complex (MHC) class II molecules on the surface of antigen-presenting cells. The class II-peptide complexes must then be recognized by T-cell receptors (TCRs). T-cell stimulation requires sustained association of antigen-presenting cells and T cells through formation of a class II-peptide-TCR complex, and peptide sequences that mediate this association are termed "T-cell epitopes." MHC class II tetramers that bind FVIII-derived peptides and recognize antigen-specific TCRs are proving useful in the characterization of human leukocyte antigen-restricted T-cell responses to FVIII. PMID- 19184560 TI - Management of haemophilia A-inhibitor patients: clinical and regulatory perspectives. AB - Inhibitors to factor VIII (FVIII) are alloantibodies directed against epitopes able to neutralise FVIII procoagulant activity. They may render FVIII replacement therapy ineffective. They represent the most severe complication of haemophilia A. At least three mechanisms of FVIII neutralisation activity by anti-FVIII antibodies have been described: (1) steric hindrance; (2) recognition of neo epitopes and (3) catalytic activity. The Nijmegen modification of the Bethesda is the recommended method for inhibitor surveillance. The occurrence of inhibitors is a relatively frequent and early event in previously untreated patients. Conversely, it is rare in previously treated patients. Therapeutic strategies for managing inhibitors include: inhibitor eradication, haemostatic management of bleeding episodes and/or surgery and supportive care. For high responding inhibitors, immune tolerance induction (ITI) is the strategy for achieving antigen-specific tolerance to FVIII. ITI success rate ranges commonly between 60% and 80%. For treatment of patients with high-titre, high-responding inhibitors, 'by-pass' therapy is generally recommended. Activated prothrombin complex concentrates represent the historically primary 'by-pass' treatment. Recombinant factor VIIa has also been widely used as a by-passing agent. Considering the small patient population, it has to be considered that full immunogenicity data cannot be collected premarketing authorisation. Thus, stringent follow-up of patients in the post-authorisation phase is required. PMID- 19184562 TI - Neuroimaging informatics tools and resources clearinghouse (NITRC) resource announcement. PMID- 19184561 TI - PyMVPA: A python toolbox for multivariate pattern analysis of fMRI data. AB - Decoding patterns of neural activity onto cognitive states is one of the central goals of functional brain imaging. Standard univariate fMRI analysis methods, which correlate cognitive and perceptual function with the blood oxygenation level dependent (BOLD) signal, have proven successful in identifying anatomical regions based on signal increases during cognitive and perceptual tasks. Recently, researchers have begun to explore new multivariate techniques that have proven to be more flexible, more reliable, and more sensitive than standard univariate analysis. Drawing on the field of statistical learning theory, these new classifier-based analysis techniques possess explanatory power that could provide new insights into the functional properties of the brain. However, unlike the wealth of software packages for univariate analyses, there are few packages that facilitate multivariate pattern classification analyses of fMRI data. Here we introduce a Python-based, cross-platform, and open-source software toolbox, called PyMVPA, for the application of classifier-based analysis techniques to fMRI datasets. PyMVPA makes use of Python's ability to access libraries written in a large variety of programming languages and computing environments to interface with the wealth of existing machine learning packages. We present the framework in this paper and provide illustrative examples on its usage, features, and programmability. PMID- 19184566 TI - Constraints to progress in embryonic stem cells from domestic species. AB - Domestic animal embryonic stem cells are of potentially big value in transgenic research and studies of lineage commitment and development. Unfortunately, despite many efforts, validated embryonic stem cell lines in species other than mice and primates are yet to be isolated. Here we review some factors that might help to explain why derivation of domestic animal embryonic stem cells is still unsuccessful. PMID- 19184565 TI - VIPoma with expression of both VIP and VPAC1 receptors in a patient with WDHA syndrome. AB - We report a case of VIPoma in a 72-year-old female patient who presented with excessive diarrhea, severe hypokalemia, and acidemia. She had been referred to our hospital three times because of severe diarrhea. No primary tumor site was found by conventional techniques, including contrast-enhanced CT and MRI, angiography, endoscopy, and positron emission tomography (PET), but a tumor was subsequently found in the head of the pancreas by octreotide scanning. Her diarrhea diminished dramatically after octreotide treatment, while her diarrhea has ceased without the therapy of octreotide at the first admission in the course of 2 years of her disease. Immunohistochemial analysis of the excised tumor tissue revealed the expression of both vasoactive intestinal peptide (VIP) and VIP and pituitary adenylate cyclase-activating peptide 1 (VPAC1) receptors. This is the first case report of a VIPoma that immunostains for VIP and VPAC1 receptors and indicates that abundant VIP produced by VIPoma might inhibit its growth and reduce VIP secretion via the VPAC1 receptor in vivo. PMID- 19184563 TI - ER stress and unfolded protein response in amyotrophic lateral sclerosis. AB - Several theories on the pathomechanism of amyotrophic lateral sclerosis (ALS) have been proposed: misfolded protein aggregates, mitochondrial dysfunction, increased glutamate toxicity, increased oxidative stress, disturbance of intracellular trafficking, and so on. In parallel, a number of drugs that have been developed to alleviate the putative key pathomechanism of ALS have been under clinical trials. Unfortunately, however, almost all studies have finished unsuccessfully. This fact indicates that the key ALS pathomechanism still remains a tough enigma. Recent studies with autopsied ALS patients and studies using mutant SOD1 (mSOD1) transgenic mice have suggested that endoplasmic reticulum (ER) stress-related toxicity may be a relevant ALS pathomechanism. Levels of ER stress-related proteins were upregulated in motor neurons in the spinal cords of ALS patients. It was also shown that mSOD1, translocated to the ER, caused ER stress in neurons in the spinal cord of mSOD1 transgenic mice. We recently reported that the newly identified ALS-causative gene, vesicle-associated membrane protein-associated protein B (VAPB), plays a pivotal role in unfolded protein response (UPR), a physiological reaction against ER stress. The ALS linked P56S mutation in VAPB nullifies the function of VAPB, resulting in motoneuronal vulnerability to ER stress. In this review, we summarize recent advances in research on the ALS pathomechanism especially addressing the putative involvement of ER stress and UPR dysfunction. PMID- 19184568 TI - Over-expression of human kallikrein 11 is associated with poor prognosis in patients with low rectal carcinoma. AB - AIM: The goal of this study was to analyze the expression of human kallikrein 11 (hK11) in low rectal carcinoma (LRC) tissues, as well as its association with the clinicopathologic features of LRC patients and its prognostic significance. METHODS: Between January 1998 and January 2003, 126 patients with LRC were randomized to receive laparoscopic-assisted abdominoperineal resection (APR). Their hK11 expression levels were examined by immunostaining on paraffin-embedded tumor specimens. Then the association of hK11 expression with clinicopathologic characteristics and patients' prognosis was analyzed. RESULTS: The positive degree of immunohistochemical staining in cancerous tissues (1.86 +/- 0.02) was higher than that in the control group (1.18 +/- 0.11, P = 0.008). Strong positive hK11 staining associated significantly with various clinicopathologic features, such as Dukes Staging (P < 0.001), histological differentiation grade (P = 0.003), CEA level (P = 0.01), lymph node metastasis (P < 0.001), and invasion depth (P = 0.002). Patients with weak hK11 positive expression showed better survival rates of 5-year follow-up than those with strong positive expression (80.85% for weak positive expression and 58.23% in strong positive expression, respectively; P < 0.01 for analyses). Using Cox regression analysis of the 126 patients, strong positive expression of hK11, Dukes Staging, lymph node metastasis, and invasion depth seemed to be independent prognostic indicators (P < 0.01, P = 0.02, P < 0.01 and P < 0.01, respectively). CONCLUSION: hK11 may be another prognostic biomarker of LRC. Knowledge of hK11 expression in LRC tissues could contribute to the prediction of prognosis of LRC patients. PMID- 19184567 TI - Pre-transplantation specification of stem cells to cardiac lineage for regeneration of cardiac tissue. AB - Myocardial infarction (MI) is a lead cause of mortality in the Western world. Treatment of acute MI is focused on restoration of antegrade flow which inhibits further tissue loss, but does not restore function to damaged tissue. Chronic therapy for injured myocardial tissue involves medical therapy that attempts to minimize pathologic remodeling of the heart. End stage therapy for chronic heart failure (CHF) involves inotropic therapy to increase surviving cardiac myocyte function or mechanical augmentation of cardiac performance. Not until the point of heart transplantation, a limited resource at best, does therapy focus on the fundamental problem of needing to replace injured tissue with new contractile tissue. In this setting, the potential for stem cell therapy has garnered significant interest for its potential to regenerate or create new contractile cardiac tissue. While to date adult stem cell therapy in clinical trials has suggested potential benefit, there is waning belief that the approaches used to date lead to regeneration of cardiac tissue. As the literature has better defined the pathways involved in cardiac differentiation, preclinical studies have suggested that stem cell pretreatment to direct stem cell differentiation prior to stem cell transplantation may be a more efficacious strategy for inducing cardiac regeneration. Here we review the available literature on pre transplantation conditioning of stem cells in an attempt to better understand stem cell behavior and their readiness in cell-based therapy for myocardial regeneration. PMID- 19184574 TI - Serving the common interest in U.S. Forest Policy: a case study of the healthy forests restoration act. AB - In the United States, the common interest often is conceived as a by-product of the pluralist, interest-group-driven democratic process. Special interests dominate in many political arenas. Consequently, we have lost the language, vocabulary, and ability to talk about the common interest. The way to reverse this trend is to develop and practice with new tools that allow us to articulate what we mean by the common interest in specific contexts. In this article, we leveraged the literature on procedural, substantive, and pragmatic decision making to illustrate how they work together to demonstrate whether and how the common interest was served in three case studies of Healthy Forests Restoration Act implementation on the Apache-Sitgreaves National Forest in Arizona. In two of the cases we found that the common interest was mostly served, while in the third case it was not. Our results raise questions about the ability of procedural criteria or substantive criteria alone to determine effectiveness in decision making. When evaluated together they provide a more complete understanding of how the common interest is or is not served. PMID- 19184573 TI - Notch signaling in cardiac development and disease. AB - The Notch pathway is an ancient, highly conserved signaling mechanism that participates in essential cell-cell communication events between adjacent cells. Mutations in Notch-signaling elements cause cardiac abnormalities in mice and humans, demonstrating an essential role for Notch in heart development. Studies with targeted mutant mice indicate that Notch signaling promotes the epithelial to-mesenchyme transition that gives rise to the cardiac valve primordium, which later is sculpted into mature valves. During ventricular chamber development, the myocardium differentiates into two layers: an outer compact zone and an inner trabecular zone. Trabeculae provide a pumping function during early phases of ventricular development and contribute to the cardiac conduction system in the mature heart. Notch regulates the endocardium-to-myocardium signals that balance proliferation and differentiation of trabecular myocytes. Recent evidence demonstrates that defective NOTCH signaling leads to aortic valve degeneration in humans. Future research will be informative about the involvement of altered NOTCH signaling in chamber abnormalities and other cardiac disorders. PMID- 19184575 TI - A comparison of sampling designs for monitoring recreational trail impacts in Rocky Mountain National Park. AB - The dual goals of the Organic Act of 1916 and Wilderness Act of 1964 are to protect natural resources and provide quality visitor experiences. Park managers need metrics of trail conditions to protect park resources and quality of visitor experiences. A few methods of sampling design for trails have been developed. Here, we describe a relatively new method, spatially balanced sampling, and compare it to systematic sampling. We evaluated the efficiency of sampling designs to measure recreation-related impacts in Rocky Mountain National Park. This study addressed two objectives: first, it compared estimates of trail conditions from data collected from systematic versus spatially balanced sampling data; second, it examined the relationship between sampling precision and sampling efficiency. No statistically significant differences in trail condition were found between the 100-m interval and the spatially balanced datasets. The spatially balanced probability-based dataset was found to be a good estimate of trail conditions when analyses were conducted with fewer sample points. Moreover, spatially balanced probability-based sampling is flexible and allows additional sample points to be added to a sample. PMID- 19184576 TI - Adaptation to climate change in developing countries. AB - Adaptation to climate change is given increasing international attention as the confidence in climate change projections is getting higher. Developing countries have specific needs for adaptation due to high vulnerabilities, and they will in this way carry a great part of the global costs of climate change although the rising atmospheric greenhouse gas concentrations are mainly the responsibility of industrialized countries. This article provides a status of climate change adaptation in developing countries. An overview of observed and projected climate change is given, and recent literature on impacts, vulnerability, and adaptation are reviewed, including the emerging focus on mainstreaming of climate change and adaptation in development plans and programs. The article also serves as an introduction to the seven research articles of this special issue on climate change adaptation in developing countries. It is concluded that although many useful steps have been taken in the direction of ensuring adequate adaptation in developing countries, much work still remains to fully understand the drivers of past adaptation efforts, the need for future adaptation, and how to mainstream climate into general development policies. PMID- 19184577 TI - Multi-destination and multi-purpose trip effects in the analysis of the demand for trips to a remote recreational site. AB - One of the basic assumptions of the travel cost method for recreational demand analysis is that the travel cost is always incurred for a single purpose recreational trip. Several studies have skirted around the issue with simplifying assumptions and dropping observations considered as nonconventional holiday makers or as nontraditional visitors from the sample. The effect of such simplifications on the benefit estimates remains conjectural. Given the remoteness of notable recreational parks, multi-destination or multi-purpose trips are not uncommon. This article examines the consequences of allocating travel costs to a recreational site when some trips were taken for purposes other than recreation and/or included visits to other recreational sites. Using a multi purpose weighting approach on data from Gros Morne National Park, Canada, we conclude that a proper correction for multi-destination or multi-purpose trip is more of what is needed to avoid potential biases in the estimated effects of the price (travel-cost) variable and of the income variable in the trip generation equation. PMID- 19184578 TI - Polymorphism in apolipoprotein E among migraineurs and tension-type headache subjects. AB - Nitric oxide plays an important role in the pathogenesis of migraine as well as tension-type headache. Studies suggest that the expression of molecules involved in the pathogenesis of headache, i.e., nitric oxide and interleukin, is influenced by apolipoprotein E (APOE) and is gene specific. Hence, we hypothesized that APOE polymorphism may be associated with migraine as well as tension-type headache.The study sample comprised of three groups: migraineurs, tension-type headache subjects as well as a healthy control group. A total of 50 subjects in each group were included after screening for the inclusion and exclusion criteria. None of the subjects was a blood relative of any other subject included in the present study. Their venous blood was drawn and stored at -20 degrees C. Genomic DNA extraction was performed with a commercial kit and simple sequence-specific primer PCR was performed to assess the APOE polymorphism. Data were analyzed with the help of SPSS V11.0 for Windows. chi(2) test and logistic regression analysis were run. The results of the study showed that APOE epsilon2 gene increases the risk of migraine as compared to the control group and the tension-type headache group (OR=4.85; 95% CI=1.92-12.72; P<0.001 and OR=2.31; 95% CI=1.08-4.94; P=0.01, respectively). Interestingly, APOE epsilon4 gene was protective against migraine as well as tension-type headache. This study shows that APOE epsilon2 gene increases the risk of migraine, while APOE epsilon4 gene is protective against migraine and tension-type headache. Further research is required to confirm the findings of the present study in a larger sample and to elucidate the role of APOE polymorphism in headache. PMID- 19184579 TI - Medulloblastoma in children under the age of three years: a retrospective Canadian review. AB - Children under the age of 3 with medulloblastoma have an inferior survival to older children with this disease. This study reviewed the incidence, characteristics, therapy, and outcome of children less than 36 months of age diagnosed with medulloblastoma from 1990 to 2005 in Canada. Ninety-six cases were identified with a median age at diagnosis of 19.5 months. Forty-seven percent of patients had a complete resection, 25% a 90-95% near complete resection, 20% an incomplete (10-90%) resection, and 3% biopsy only. Therapy consisted of chemotherapy (90%), high dose chemotherapy with stem cell rescue (13%), and radiation therapy (21%). The median survival time was 45 +/- 13.82 months. There was no significant difference in survival when comparing patients with <90% resection versus >90% resection, nor when comparing the presence of metastases versus their absence. There was a significant increase in survival time in patients who received radiation therapy compared to those who were not treated with this modality, as well as for those who were over 18 months at diagnosis compared to those under 18 months. PMID- 19184580 TI - Molecular analysis of Ixodes granulatus, a possible vector tick for Borrelia burgdorferi sensu lato in Taiwan. AB - The genetic identity of Ixodes granulatus ticks was determined for the first time in Taiwan. The phylogenetic relationships were analyzed by comparing the sequences of mitochondrial 16S ribosomal DNA gene obtained from 19 strains of ticks representing seven species of Ixodes and two outgroup species (Rhipicephalus sanguineus and Haemaphysalis inermis). Four major clades could be easily distinguished by neighbour-joining analysis and were congruent by maximum parsimony method. All these I. granulatus ticks of Taiwan were genetically affiliated to a monophyletic group with highly homogeneous sequences (92.2-99.3% similarity), and can be discriminated from other Ixodes species and other genera of ticks with a sequence divergence ranging from 11.7 to 30.8%. Moreover, intraspecific analysis revealed that two distinct lineages are evident between the same species of I. granulatus ticks collected from Taiwan and Malaysia. Our results demonstrate that all these I. granulatus ticks of Taiwan represent a unique lineage distinct from the common vector ticks (I. ricinus complex) for Borrelia burgdorferi spirochetes. PMID- 19184581 TI - Oxalic acid: a prospective tool for reducing Varroa mite populations in package bees. AB - Numerous studies have investigated using oxalic acid (OA) to control Varroa mites in honey bee colonies. In contrast, techniques for treating package bees with OA have not been investigated. The goal of this study was to develop a protocol for using OA to reduce mite infestation in package bees. We made 97 mini packages of Varroa-infested adult bees. Each package contained 1,613 +/- 18 bees and 92 +/- 3 mites, and represented an experimental unit. We prepared a 2.8% solution of OA by mixing 35 g OA with 1 l of sugar water (sugar:water = 1:1; w:w). Eight treatments were assigned to the packages based on previous laboratory bioassays that characterized the acute contact toxicity of OA to mites and bees. We administered the treatments by spraying the OA solution directly on the bees through the mesh screen cage using a pressurized air brush and quantified mite and bee mortality over a 10-day period. Our results support applying an optimum volume of 3.0 ml of a 2.8% OA solution per 1,000 bees to packages for effective mite control with minimal adult bee mortality. The outcome of our research provides beekeepers and package bee shippers guidance for using OA to reduce mite populations in package bees. PMID- 19184583 TI - Marine toxins: an overview. AB - Oceans provide enormous and diverse space for marine life. Invertebrates are conspicuous inhabitants in certain zones such as the intertidal; many are soft bodied, relatively immobile and lack obvious physical defenses. These animals frequently have evolved chemical defenses against predators and overgrowth by fouling organisms. Marine animals may accumulate and use a variety of toxins from prey organisms and from symbiotic microorganisms for their own purposes. Thus, toxic animals are particularly abundant in the oceans. The toxins vary from small molecules to high molecular weight proteins and display unique chemical and biological features of scientific interest. Many of these substances can serve as useful research tools or molecular models for the design of new drugs and pesticides. This chapter provides an initial survey of these toxins and their salient properties. PMID- 19184582 TI - Regulation of immunity at tissue sites of inflammation. AB - The acquisition and execution of CD4 effector function are tightly regulated and spatially compartmentalized. In the lymph node (LN), naive CD4+ T cells acquire specialized functions by means of expression of distinct cytokines and acquire distinct homing properties. Therefore, both the function and subsequent localization of effector cells appears to be predetermined during differentiation in the LN. Our studies with the protozoa Leishmania major suggest that this centrally (LN) generated effector repertoire can be further edited at the infected tissue site. Cytokine production in the inflamed tissue can be modulated at a number of levels including chemokine-driven differential recruitment of effector cells, the provision of signals for effector cell function and suppression by regulatory T cells (Tregs). The concept that tissue resident pathogens may subvert the centrally generated cytokine repertoire has important therapeutic implications. Novel therapies that focus on manipulating the local infection site to encourage appropriate recruitment or activation of effectors may be particularly beneficial. PMID- 19184584 TI - Conotoxins: molecular and therapeutic targets. AB - Marine molluscs known as cone snails produce beautiful shells and a complex array of over 50,000 venom peptides evolved for prey capture and defence. Many of these peptides selectively modulate ion channels and transporters, making them a valuable source of new ligands for studying the role these targets play in normal and disease physiology. A number of conopeptides reduce pain in animal models, and several are now in pre-clinical and clinical development for the treatment of severe pain often associated with diseases such as cancer. Less than 1% of cone snail venom peptides are pharmacologically characterised. PMID- 19184585 TI - Sodium channel inhibiting marine toxins. AB - Saxitoxin (STX), tetrodotoxin (TTX) and their many chemical relatives are part of our daily lives. From killing people who eat seafood containing these toxins, to being valuable research tools unveiling the invisible structures of their pharmacological receptor, their global impact is beyond measure. The pharmacological receptor for these toxins is the voltage-gated sodium channel which transports Na ions between the exterior to the interior of cells. The two structurally divergent families of STX and TTX analogues bind at the same location on these Na channels to stop the flow of ions. This can affect nerves, muscles and biological senses of most animals. It is through these and other toxins that we have developed much of our fundamental understanding of the Na channel and its part in generating action potentials in excitable cells. PMID- 19184586 TI - Sea anemone toxins affecting potassium channels. AB - The great diversity of K(+) channels and their wide distribution in many tissues are associated with important functions in cardiac and neuronal excitability that are now better understood thanks to the discovery of animal toxins. During the past few decades, sea anemones have provided a variety of toxins acting on voltage-sensitive sodium and, more recently, potassium channels. Currently there are three major structural groups of sea anemone K(+) channel (SAK) toxins that have been characterized. Radioligand binding and electrophysiological experiments revealed that each group contains peptides displaying selective activities for different subfamilies of K(+) channels. Short (35-37 amino acids) peptides in the group I display pore blocking effects on Kv1 channels. Molecular interactions of SAK-I toxins, important for activity and binding on Kv1 channels, implicate a spot of three conserved amino acid residues (Ser, Lys, Tyr) surrounded by other less conserved residues. Long (58-59 amino acids) SAK-II peptides display both enzymatic and K(+) channel inhibitory activities. Medium size (42-43 amino acid) SAK-III peptides are gating modifiers which interact either with cardiac HERG or Kv3 channels by altering their voltage-dependent properties. SAK-III toxins bind to the S3C region in the outer vestibule of Kv channels. Sea anemones have proven to be a rich source of pharmacological tools, and some of the SAK toxins are now useful drugs for the diagnosis and treatment of autoimmune diseases. PMID- 19184589 TI - Toxins affecting actin filaments and microtubules. AB - Actin and tubulin are the two major proteins of the cytoskeleton in eukaryotic cells and both display a common property to reversibly assemble into long and flexible polymers, actin filaments and microtubules, respectively. These proteins play important roles in a variety of cellular functions and are also involved in numbers of diseases. An emerging number of marine-derived cytotoxins have been found to bind either actin or tublin, resulting in either inhibition or enhancement of polymerization. Thus, these toxins are valuable molecular probes for solving complex mechanisms of biological processes. This chapter describes actin- and tubulin-targeting marine natural products and their modes of action, with reference to their use as research tools and their clinical applications. PMID- 19184588 TI - Marine toxins potently affecting neurotransmitter release. AB - Synapses are specialised structures where interneuronal communication takes place. Not only brain function is absolutely dependent on synaptic activity, but also most of our organs are intimately controlled by synaptic activity. Synapses re therefore an ideal target to act upon and poisonous species have evolved fascinating neurotoxins capable of shutting down neuronal communication by blocking or activating essential components of the synapse. By hijacking key proteins of the communication machinery, neurotoxins are therefore extremely valuable tools that have, in turn, greatly helped our understanding of synaptic biology. Moreover, analysis and understanding of the molecular strategy used by certain neurotoxins has allowed the design of entirely new classes of drugs acting on specific targets with high selectivity and efficacy. This chapter will discuss the different classes of marine neurotoxins, their effects on neurotransmitter release and how they act to incapacitate key steps in the process leading to synaptic vesicle fusion. PMID- 19184587 TI - Ligands for ionotropic glutamate receptors. AB - Marine-derived small molecules and peptides have played a central role in elaborating pharmacological specificities and neuronal functions of mammalian ionotropic glutamate receptors (iGluRs), the primary mediators of excitatory synaptic transmission in the central nervous system (CNS). As well, the pathological sequelae elicited by one class of compounds (the kainoids) constitute a widely-used animal model for human mesial temporal lobe epilepsy (mTLE). New and existing molecules could prove useful as lead compounds for the development of therapeutics for neuropathologies that have aberrant glutamatergic signaling as a central component. In this chapter we discuss natural source origins and pharmacological activities of those marine compounds that target ionotropic glutamate receptors. PMID- 19184590 TI - Carcinogenic aspects of protein phosphatase 1 and 2A inhibitors. AB - Okadaic acid is functionally a potent tumor promoter working through inhibition of protein phosphatases 1 and 2A (PP1 and PP2A), resulting in sustained phosphorylation of proteins in cells. The mechanism of tumor promotion with okadaic acid is thus completely different from that of the classic tumor promoter phorbol ester. Other potent inhibitors of PP1 and PP2A - such as dinophysistoxin 1, calyculins A-H, microcystin-LR and its derivatives, and nodularin - were isolated from marine organisms, and their structural features including the crystal structure of the PP1-inhibitor complex, tumor promoting activities, and biochemical and biological effects, are here reviewed. The compounds induced tumor promoting activity in three different organs, including mouse skin, rat glandular stomach and rat liver, initiated with three different carcinogens. The results indicate that inhibition of PP1 and PP2A is a general mechanism of tumor promotion applicable to various organs. This study supports the concept of endogenous tumor promoters in human cancer development. PMID- 19184591 TI - The femoral sulcus in total knee arthroplasty. AB - The position of the femoral sulcus relative to the midline of the distal femoral resection in total knee arthroplasty (TKA) was studied to determine if centralized placement of the femoral component on the distal femur was justified in terms of aligning the prosthetic sulcus with the native femoral sulcus. The location of the femoral sulcus was studied in 112 consecutive patients undergoing TKA. The mean sulcus position was 0.7 mm lateral to the midline of the distal femoral resection (SD 1.4, 95% CI, 0.5-1.0 mm). However, the variation in sulcus positions ranged from 4 mm medial to 4 mm lateral to the midline. The mean sulcus position in valgus knees was 1.0 mm lateral to the midline (SD 1.8), and that in varus knees was 0.7 mm lateral to the midline (SD 1.2) (P = 0.501). It appears prudent to centre the femoral component on the native sulcus rather than the midline of the distal femoral resection, so as to ensure accurate alignment of the prosthetic sulcus with the native sulcus and to encourage normal patella tracking. PMID- 19184593 TI - Determination of benzimidazole fungicides in water samples by on-line MISPE-HPLC. AB - An analytical methodology based on an on-line sample enrichment of water samples by means of an imprinted polymer, and the separation of benzimidazole compounds within a C(18) column by ion-pair reversed-phase liquid chromatography, has been developed. The molecularly imprinted polymer has been synthesized by precipitation polymerization using thiabendazole as template molecule, methacrylic acid as functional monomer, and divinylbenzene as cross-linker. Initial experiments carried out by solid-phase extraction on cartridges demonstrated a clear imprint effect for thiabendazole, as well as the ability of the imprinted polymer to selectively rebind several benzimidazole compounds. The developed methodology has been applied to the quantification of thiabendazole, carbendazim, and benomyl in river, tap, and well water samples within a single analytical run at concentration levels below the legislated maximum concentration levels. In this sense, detection limits of 2.3-5.7 ng.L(-1) have been obtained for the analysis of benzimidazole fungicides in different water matrices. Recoveries obtained for the determination of benzimidazole fungicides in spiked samples ranged from 87% to 95%, with RSD below 5% in all cases. PMID- 19184592 TI - A contribution to Pisani's sign for diagnosing lateral meniscal cysts: a technical report. AB - This article describes modified physical examination maneuvers for diagnosing lateral meniscal cysts. The typical presentation of lateral meniscal cysts usually makes their clinical diagnosis simple, but a wide variation in size may sometimes lead to misdiagnosis. We evaluated the effect of leg rotations on the cystic swelling at 45 degrees of knee flexion. Eleven consecutive patients with positive Pisani's sign were evaluated prospectively. Lateral mid-third joint-line swelling was most prominent at 30-45 degrees of knee flexion. With the knee held at 45 degrees of flexion, the prominence was also inspected during internal and external rotation of the leg. The masses became noticeably more prominent with external rotation, and completely disappeared with internal rotation. With external rotation of the leg, even doubtful lateral meniscal cysts became apparent. Disappearance of the cystic mass with internal rotation further confirms the diagnosis. PMID- 19184594 TI - Two-step liquid-phase microextraction and high-performance liquid chromatography for the simultaneous analysis of the enantiomers of mefloquine and its main metabolite carboxymefloquine in plasma. AB - A method for the simultaneous analysis of the enantiomers of mefloquine (MQ) and its main metabolite carboxymefloquine (CMQ) in plasma is described for the first time. The assay involves two-step liquid-phase microextraction (LPME) and enantioselective high-performance liquid chromatography. In the first LPME step, the enantiomers of MQ were extracted from an alkalinized sample through a thin layer of di-n-hexyl ether immobilized in the pores of the hollow fiber and into 0.01 M perchloric acid as acceptor solution. In the second LPME step, the same sample was acidified to enable the extraction of CMQ using the same organic solvent and 0.05 M sodium hydroxide as acceptor phase. The analytes were resolved on a Chirobiotic T column in the polar-organic mode of elution and detected at 285 nm. The recovery rates from 1 mL of plasma were in the range 35-38%. The method presented limits of quantification of 50 ng/mL for all analytes and was linear up to 1,500 and 3,000 ng/mL for the enantiomers of MQ and CMQ, respectively. The plasmatic concentrations of (+)-(RS)-MQ were higher than those of (-)-(SR)-MQ after oral administration of the racemic drug to rats. PMID- 19184595 TI - Production of oligosaccharides and cellobionic acid by Fibrobacter succinogenes S85 growing on sugars, cellulose and wheat straw. AB - Extracellular culture fluid of Fibrobacter succinogenes S85 grown on glucose, cellobiose, cellulose or wheat straw was analysed by 2D-NMR spectroscopy. Cellodextrins did not accumulate in the culture medium of cells grown on cellulose or straw. Maltodextrins and maltodextrin-1P were identified in the culture medium of glucose, cellobiose and cellulose grown cells. New glucose derivatives were identified in the culture fluid under all the substrate conditions. In particular, a compound identified as cellobionic acid accumulated at high levels in the medium of F. succinogenes S85 cultures. The production of cellobionic acid (and cellobionolactone also identified) was very surprising in an anaerobic bacterium. The results suggest metabolic shifts when cells were growing on solid substrate cellulose or straw compared to soluble sugars. PMID- 19184596 TI - Dry anaerobic ammonia-methane production from chicken manure. AB - The effect of temperature on production of ammonia during dry anaerobic fermentation of chicken manure (CM), inoculated with thermophilic methanogenic sludge, was investigated in a batch condition for 8 days. Incubation temperature did not have a significant effect on the production of ammonia. Almost complete inhibition of production of methane occurred at 55 and 65 degrees C while quite low yields of 8.45 and 6.34 ml g(-1) VS (volatile solids) were observed at 35 and 45 degrees C due to a higher accumulation of ammonia. In order to improve the production of methane during dry anaerobic digestion of CM, stripping of ammonia was performed firstly on the CM previously fermented at 65 degrees C for 8 days: the stripping for 1 day at 85 degrees C and pH 10 removed 85.5% of ammonia. The first-batch fermentation of methane for 75 days was conducted next, using the ammonia-stripped CM inoculated with methanogenic sludge at different ratios, (CM: thermophilic sludge) of 1:2, 1:1, and 2:1 on volume per volume basis at both 35 and 55 degrees C. Production of methane improved and was higher than that of the control (without stripping of ammonia) but the yield of 20.4 ml g(-1) VS was still low, so second stripping of ammonia was conducted, which resulted in 74.7% removal of ammonia. A great improvement in the production of methane of 103.5 ml g(-1) VS was achieved during the second batch for 55 days. PMID- 19184597 TI - Metabolic effects of furaldehydes and impacts on biotechnological processes. AB - There is a growing awareness that lignocellulose will be a major raw material for production of both fuel and chemicals in the coming decades--most likely through various fermentation routes. Considerable attention has been given to the problem of finding efficient means of separating the major constituents in lignocellulose (i.e., lignin, hemicellulose, and cellulose) and to efficiently hydrolyze the carbohydrate parts into sugars. In these processes, by-products will inevitably form to some extent, and these will have to be dealt with in the ensuing microbial processes. One group of compounds in this category is the furaldehydes. 2-Furaldehyde (furfural) and substituted 2-furaldehydes--most importantly 5 hydroxymethyl-2-furaldehyde--are the dominant inhibitory compounds found in lignocellulosic hydrolyzates. The furaldehydes are known to have biological effects and act as inhibitors in fermentation processes. The effects of these compounds will therefore have to be considered in the design of biotechnological processes using lignocellulose. In this short review, we take a look at known metabolic effects, as well as strategies to overcome problems in biotechnological applications caused by furaldehydes. PMID- 19184599 TI - Health disparities among immigrant and non-immigrant elders: the association of acculturation and education. AB - Guided by the theories of human capital and acculturation, this study investigated the association of immigrant status among older people with their physical and mental health outcomes, health services utilization, and health insurance coverage. Specifically, it examined the interactive effects of immigrant status, education, acculturation, race, and ethnicity on these dependent variables. The study used a national representation sample of 7,345 older Americans from the first wave of the Asset and Health Dynamic of the Oldest Old study (AHEAD) survey. We used both logistic regression and ordered logit regression for our multivariate analyses. The findings are as follows: (1) immigrant status was negatively associated with level of depression, number of IADL difficulties, and on types of health insurance coverage. Immigrant status had a significant relationship only with the utilization of outpatient surgery, but not on other health services utilization. (2) There were significant interactive effects of race and ethnicity and immigrant status on these dependent variables. The findings support the existence of double jeopardy among those who are simultaneously an immigrant and a member of a racial and ethnic minority group in the United States. (3) Acculturation has strong associations with health insurance coverage and with number of difficulties with IADL. PMID- 19184600 TI - HLA ligand profiles of primary renal cell carcinoma maintained in metastases. AB - In recent years, several approaches have been taken in the peptide-based immunotherapy of metastatic renal cell carcinoma (RCC), although little is known about HLA presentation on metastases compared to primary tumor and normal tissue of RCC. In this study we compared primary tumor, normal tissue and metastases with the aim of identifying similarities and differences between these tissues. We performed this comparison for two RCC patients on the level of the HLA ligandome using mass spectrometry and for three patients on the level of the transcriptome using oligonucleotide microarrays. The quantitative results show that primary tumor is more similar to metastasis than to normal tissue, both on the level of HLA ligand presentation and mRNA. We were able to characterize a total of 142 peptides in the qualitative analysis of HLA-presented peptides. Six of them were significantly overpresented on metastasis, among them a peptide derived from CD151; fourteen were overpresented on both primary tumor and metastasis compared to normal tissue, among them an HLA ligand derived from tumor protein p53. Thus, we could demonstrate that peptide-based immunotherapy might affect tumor as well as metastasis of RCC, but not healthy kidney tissue. Furthermore we were able to identify several peptides derived from tumor associated antigens that are suitable for vaccination of metastatic RCC. PMID- 19184601 TI - Phase II study of an all-oral combination of vinorelbine with capecitabine in patients with metastatic breast cancer. AB - PURPOSE: Combination of intravenous (i.v.) vinorelbine and capecitabine was shown to be feasible and effective in metastatic breast cancer (MBC). In an effort to improve patient convenience and to prolong infusion-free survival, we investigated in first-line treatment a regimen combining oral vinorelbine and capecitabine in a phase II study. PATIENTS AND METHODS: Fifty-two patients (median age, 60 years) with MBC received the combination consisting of oral vinorelbine 60 mg/m(2) on days 1, 8 and 15 plus capecitabine 1,000 mg/m(2) bid given from day 1 to day 14 in an open-label, multicentre phase II study [the recommended doses were established in a phase I study (Nole et al. in Ann Oncol 17:332-339, 2006)]. Cycles were repeated every 3 weeks. RESULTS: Seventy-nine percent of the patients had received prior adjuvant chemotherapy and 81% presented with visceral involvement. The median number of administered cycles per patient was 7 (range 1-18). Twenty-three responses were documented and validated by an independent panel review, yielding response rates of 44.2% (95% CI, 30.5 58.7) in the 52 enrolled patients and 54.8% (95% CI, 38.7-70.2) in the 42 evaluable patients. Median progression-free survival and median overall survival were 8.4 and 25.8 months, respectively. Neutropenia was the main dose-limiting toxicity but complications were uncommon, only one patient having experienced febrile neutropenia. Other frequently reported adverse events included, fatigue, nausea, vomiting, diarrhoea and constipation, stomatitis and hand-foot syndrome, which were rarely severe. CONCLUSIONS: This regimen combining oral vinorelbine with capecitabine is effective and manageable in the first-line treatment of MBC. Oral vinorelbine on days 1, 8 and 15 with capecitabine from days 1 to 14 every 3 weeks represents a convenient option which offers an all-oral treatment to the patients and prolongs their infusion-free survival. PMID- 19184602 TI - [Genetic causes of hereditary cone and cone-rod dystrophies]. AB - BACKGROUND: Cone and cone-rod dystrophies belong to the genetically and phenotypically very heterogeneous group of retinal degenerations. This article aims to review the current knowledge of genes and mutations involved in these rare disorders that primarily affect the cone photoreceptor system. METHODS: Literature and database search and summary of our own molecular genetic analyses in patients affected by achromatopsia, blue cone monochromatism, and cone and cone-rod dystrophy. RESULTS: Cone and cone-rod dystrophies can be divided according to the disease course into stationary and progressive disorders or by the genetic mode of inheritance into autosomal-recessive, autosomal-dominant, and X-linked traits. To date, seven genes for autosomal-recessive and nine for autosomal-dominant inherited forms of cone and cone-rod dystrophy, as well as two underlying genes on the X chromosome, have been identified. Linkage analyses imply two additional loci for autosomal-dominant, autosomal-recessive, and X linked forms of these disorders. CONCLUSION: Reliable data on the prevalence and incidence of hereditary cone and cone-rod dystrophies and the underlying genetic defects exist only for distinct clinical and genetic entities. Analysis of the known genes results in identification of the genetic defect and mutation in only a subset of patients. PMID- 19184603 TI - Applications for fees waivers on medical and psychiatric grounds at an Irish third-level college. AB - BACKGROUND: Physical and mental illness present substantial challenges to college students and authorities. AIMS: (1) To characterize medical and psychiatric reasons underlying requests for fees waivers at an Irish third-level college; (2) to identify information included in applications; and (3) to develop proposals for standardizing applications. METHODS: We examined all applications for exemption from fees for medical or psychiatric reasons over two academic years at an Irish third-level college. RESULTS: Two hundred and twenty-two students applied for exemptions; 91% were successful. Psychiatric problems were more common than physical illness (64.1 vs. 27.9%). There was wide variation in details provided in applications with few providing details on likely ongoing duration of illness (5.0%). CONCLUSIONS: We recommend that (a) a structured medical application form be completed by medical practitioners supporting applications; (b) a medical practitioner review applications; and (c) there should be annual re-audit of this process. PMID- 19184604 TI - Metastatic uterine leiomyosarcoma presenting as a breast lump. AB - BACKGROUND: It is uncommon to encounter a breast metastasis from an extramammary malignancy and even rarer from a uterine leiomyosarcoma. AIMS: We describe the third case report in the medical literature of a breast metastasis from a uterine leiomyosarcoma. METHODS: We report the management of a 56-year-old patient who presented with a breast lump 3 years after hysterectomy for a fibroid uterus. We conducted a literature review of breast leiomyosarcomas. RESULTS: The excision of the breast mass revealed a low-grade leiomyosarcoma. Radiographic examinations demonstrated metastases to the lung, liver, pelvis and bone. Retrospective pathology review of her uterus identified a small focus of leiomyosarcoma. She received chemotherapy and palliative radiotherapy but passed away within few months. CONCLUSION: Metastasis to the breast from a non-breast primary is generally a sign of disseminated disease and; thus, a poor prognostic indicator. PMID- 19184605 TI - The role of transthoracic echocardiography in embolic acute limb ischaemia. AB - INTRODUCTION: Embolic acute limb ischaemia (ALI) is commonly treated by re vascularization and long-term anticoagulant therapy. Transthoracic echocardiography (TTE) is commonly used to screen for cardiac embolic source, but may not affect management. REPORT: We reviewed 115 consecutive patients with embolic ALI, 61% underwent TTE, with cardiac thrombus identified in only 3%. Incidental severe abnormalities requiring further cardiological investigation were detected in 19% of patients. Inpatient TTE did not affect mortality, morbidity, amputation rate, or anticoagulation. DISCUSSION: Transthoracic echocardiography seldom identifies a cardiac embolic source, but identifies many patients with severe incidental cardiac abnormalities, suggesting cardiology screening of these patients remains important. PMID- 19184607 TI - Sarcoidosis presenting with severe hypocalcaemia. AB - Disorders of calcium metabolism, especially hypercalcemia and hypercalceuria, are common in sarcoidosis. They are caused by extra renal unsuppressed production of 1,25 dihydroxy vitamin D at the level of the sarcoid granuloma. Hormonal changes during pregnancy have a physiological synergistic effect on this mechanism, which is primarily parathyroid hormone (PTH) dependant. However, the combination of primary hypoparathyroidism with hypocalcemia and sarcoidosis is rare. Di George syndrome, is a dysmorphic disorder characterised by aplasia/hypoplasia of thymus and parathyroid glands in addition to aortic arch anamolies and facial dysmorphia. After commencing appropriate treatment this lady made excellent recovary. PMID- 19184608 TI - Comment on Shuhaibar et al.: Favourable effect of immunomodulator therapy on bone mineral density in multiple sclerosis. PMID- 19184609 TI - Monitoring oxidative stress and DNA damage induced by heavy metals in yeast expressing a redox-sensitive green fluorescent protein. AB - Reactive oxygen species (ROS) has been proposed to play an important role in heavy metal-associated toxicity and pathology. Conventional methods for determining ambient redox state in cells are usually labor-intensive, precluding real-time or single-cell monitoring changes in intracellular redox poise resulting from either metabolic processes or environmental influences. Redox sensitive green fluorescent protein (roGFP) was expressed in Saccharomyces cerevisiae and recombinant cells were evaluated in monitoring the changes in the redox state of living cells when challenged with toxicologically relevant metal ions. roGFP expressed in yeast responded not only to typical membrane-permeant oxidants and reductants, but also to toxicological metal ion-induced intracellular redox changes. Moreover, exposure of yeast cells to NaAsO(2) or Pb(NO(3))(2) at concentrations that induced redox changes reported by roGFP caused up to two- to three-fold increases in DNA mutation frequency. This mutagenic effect was largely caused by oxidative stress since blocking the production of hydryl radicals significantly reduced the mutation rate as well as delayed the cell death. PMID- 19184610 TI - Extracellular hemicellulolytic enzymes from the maize endophyte Acremonium zeae. AB - Microorganisms that colonize plants require a number of hydrolytic enzymes to help degrade the cell wall. The maize endophyte Acremonium zeae was surveyed for production of extracellular enzymes that hydrolyze cellulose and hemicellulose. The most prominent enzyme activity in cell-free culture medium from A. zeae NRRL 6415 was xylanase, with a specific activity of 60 U/mg from cultures grown on crude corn fiber. Zymogram analysis following SDS-PAGE indicated six functional xylanase polypeptides of the following masses: 51, 44, 34, 29, 23, and 20 kDa. Xylosidase (0.39 U/mg), arabinofuranosidase (1.2 U/mg), endoglucanase (2.3 U/mg), cellobiohydrolase (1.3 U/mg), and beta-glucosidase (0.85 U/mg) activities were also detected. Although apparently possessing a full complement of hemicellulolytic activities, cell-free culture supernatants prepared from A. zeae required an exogenously added xylosidase to release more than 90% of the xylose and 80% of the arabinose from corn cob and wheat arabinoxylans. The hydrolytic enzymes from A. zeae may be suitable for application in the bioconversion of lignocellulosic biomass into fermentable sugars. PMID- 19184611 TI - Low common bile duct bifurcation incidentally discovered during pancreaticoduodenectomy. AB - INTRODUCTION: Bile duct injury due to failure to recognize anatomical variations can have considerable consequences. DISCUSSION: We report an incidental discovery of a low common bile duct bifurcation below the level of the cystic duct, incidentally discovered during pancreaticoduodenectomy. PMID- 19184612 TI - Risk factors of survival and surgical treatment for advanced gastric cancer with large tumor size. AB - BACKGROUND: The purpose of this study was to clarify the clinical significance of tumor size in advanced gastric cancer and to evaluate the risk factors of survival in advanced gastric cancer with large tumor size. METHODS: The cut-off point for tumor size, 90th percentile value of tumor size in advanced gastric cancer, was determined to be 10 cm. We retrospectively studied the clinicopathological features and prognosis of 406 patients with advanced gastric tumors measuring 10 cm or more. RESULTS: Large tumors had a propensity for the following: Borrmann type IV, adjacent organ invasion, lymph node and distant metastasis, and stage IV classification. Tumor size was an independent risk factor for lymph node metastasis and survival in advanced gastric cancer. In patients with large advanced gastric cancer, Borrmann type IV, adjacent organ invasion, and N2-3 nodal involvement were independent factors associated with a poorer prognosis. The 5-year survival rate in large gastric cancer patients without any risk factors (65.5%) was similar with those in small gastric cancer patients (59.3%, P = 0.123). CONCLUSION: Tumor size was a simple predictor for lymph node metastasis and survival in advanced gastric cancer. Radical surgery should be recommended for large advanced gastric cancer patients without risk factors, while large gastric cancer with risk factors may not be a surgically treatable disease. PMID- 19184613 TI - Hemorrhage-induced hepatic injury and hypoperfusion can be prevented by direct peritoneal resuscitation. AB - BACKGROUND: Crystalloid fluid resuscitation after hemorrhagic shock (HS) that restores/maintains central hemodynamics often culminates in multi-system organ failure and death due to persistent/progressive splanchnic hypoperfusion and end organ damage. Adjunctive direct peritoneal resuscitation (DPR) using peritoneal dialysis solution reverses HS-induced splanchnic hypoperfusion and improves survival. We examined HS-mediated hepatic perfusion (galactose clearance), tissue injury (histopathology), and dysfunction (liver enzymes). METHODS: Anesthetized rats were randomly assigned (n = 8/group): (1) sham (no HS); (2) HS (40% mean arterial pressure for 60 min) plus conventional i.v. fluid resuscitation (CR; shed blood + 2 volumes saline); (3) HS + CR + 30 mL intraperitoneal (IP) DPR; or (4) HS + CR + 30 mL IP saline. Hemodynamics and hepatic blood flow were measured for 2 h after CR completion. In duplicate animals, liver and splanchnic tissues were harvested for histopathology (blinded, graded), hepatocellular function (liver enzymes), and tissue edema (wet-dry ratio). RESULTS: Group 2 decreased liver blood flow, caused liver injuries (focal to submassive necrosis, zones 2 and 3) and tissue edema, and elevated liver enzymes (alanine aminotransferase (ALT), 149 +/- 28 microg/mL and aspartate aminotransferase (AST), 234 +/- 24 microg/mL; p < 0.05) compared to group 1 (73 +/- 9 and 119 +/- 10 microg/mL, respectively). Minimal/no injuries were observed in group 3; enzymes were normalized (ALT 89 +/- 9 microg/mL and AST 150 +/- 17 microg/mL), and tissue edema was similar to sham. CONCLUSIONS: CR from HS restored and maintained central hemodynamics but did not restore or maintain liver perfusion and was associated with significant hepatocellular injury and dysfunction. DPR added to conventional resuscitation (blood and crystalloid) restored and maintained liver perfusion, prevented hepatocellular injury and edema, and preserved liver function. PMID- 19184614 TI - Synchronous intussusceptions following Roux-en-Y Gastric Bypass: case report and review of the literature. AB - Intussusception is an atypical cause of small bowel obstruction following Roux-en Y gastric bypass and, as such, is not often considered in the differential diagnosis. In this setting, the pathophysiology appears to differ in that a pathologic lead point is rarely identified and the intussusceptum most often progresses retrograde into the proximal bowel. Involvement of the jejunojejunostomy is nearly universal in reported cases and, if untreated, can lead to devastating consequences. Despite this potential, the clinical presentation often lacks remarkable features yet timely surgical intervention is generally required. This phenomenon is being reported with greater frequency and altered motility in the Roux limb is the prevailing hypothesis regarding pathogenesis. Awareness of the potential for this process and prompt consultation with a bariatric surgeon are keys to optimal outcomes. PMID- 19184615 TI - Manufacturers affect clinical results of THA with zirconia heads: a systematic review. AB - In the 1980s, zirconia was introduced for THA with the expectation of lower polyethylene wear and better clinical results. However, several studies have reported poor survivorship of zirconia-polyethylene THA. We performed a systematic review and meta-analysis of zirconia-polyethylene THA to confirm or refute the theoretical advantages of this combination. Of 163 studies identified by a comprehensive search, seven met our selection criteria. These involved 769 hips of 586 patients with a mean age of 56.8 years and a minimum followup of 60 months (mean, 89.2 months; range, 60-155 months). The consolidated revision rate of zirconia-polyethylene THA at 89.2 months was higher than that of nonzirconia polyethylene THA by 5% (risk difference, 0.05; 95% confidence interval, 0.02 0.08). Subgroup meta-analysis suggested THAs with zirconia heads from Ceraver had more revision surgery than nonzirconia heads (risk difference, 0.08; 95% confidence interval, 0.03-0.14), whereas zirconia heads from DePuy did not (risk difference, 0.02; 95% confidence interval, -0.01-0.06). The meta-analysis for annual linear polyethylene wear (which did not involve zirconia heads from Ceraver because of insufficient descriptions) showed no difference between zirconia and control groups. Collectively, THAs with high-quality zirconia heads appear to have prosthesis survivorship and polyethylene wear equivalent to those of THAs with traditional materials, but differing quality among zirconia heads could lead to poor survivorship of prostheses. PMID- 19184616 TI - Aldehyde dehydrogenase activity allows reliable EPC enumeration in stored peripheral blood samples. AB - BACKGROUND: Interest in the biology of endogenous progenitor cells (EPCs) continues to grow as evidence of their role in vascular repair mounts. EPC enumeration requires specialized laboratory techniques and is performed immediately after sample acquisition, limiting the clinical contexts in which EPC enumeration can be performed and the ability to increase sample sizes through multi-center participation. METHODS: We compared the numbers of EPCs enumerated in samples processed immediately after acquisition (n = 36) with EPCs enumerated in specimens stored for 24 hours or after cryopreservation of mononuclear cells (MNC) using two EPC identification strategies: cell surface marker expression (CD133/CD34) and aldehyde dehydrogenase activity (ALDH(br) cells). RESULTS: EPCs assessed in fresh samples correlated with EPCs enumerated after whole blood storage (r = 0.699 for CD133(+)CD34(+) cells, r = 0.880 for ALDH(br) cells, P < 0.005 and P < 0.0001, respectively) or mononuclear cryopreservation (r = 0.590 for CD133(+)CD34(+) cells, r = 0.894 for ALDH(br) cells, P < 0.0001 for each); however, correlation based on assessment of ALDH(br) cells was higher (P < 0.0003 for comparison of correlation coefficients). Initial results from a multi-site clinical trial suggest that EPC enumeration after mononuclear cell cryopreservation is feasible. CONCLUSION: EPC analysis based on ALDH activity is reproducible, even after extended whole blood storage or MNC cryopreservation. PMID- 19184617 TI - Pulmonary spray dried powders of tobramycin containing sodium stearate to improve aerosolization efficiency. AB - PURPOSE: Tobramycin microparticulate powders containing the hydrophobic adjunct sodium stearate were studied for their use as pulmonary formulations in dry powder inhalers. METHODS: Spray-dried powders were characterized in terms of particle size distribution, morphology, crystallinity, drug dissolution rate, toxicity on epithelial lung cells and aerosol efficiency. RESULTS: The presence of the sodium stearate had a direct influence on the aerosol performance of tobramycin spray-dried powders. Powders containing 1% w/w sodium stearate had fine particle fraction FPF of 84.3 +/- 2.0% compared to 27.1 +/- 1.9% for powders containing no adjunct. This was attributed to the accumulation of sodium stearate at the particle surface. Powders with higher sodium stearate concentrations (2% w/w) showed significantly lower FPF (66.4 +/- 0.9%) and less accumulation of sodium stearate at the particle surface. This was attributed to the formation of adjunct micelles, which remained internalised in the particle structure due to their reduced tropism toward the drying drop surface and molecular mobility. Preliminary analysis of the toxicity effect of sodium stearate on A549 cell lines showed that the adjunct, in the concentration used, had no effect on cell viability over a 24-h period compared to particles of pure tobramycin. CONCLUSIONS: Tobramycin pulmonary powders with low level of sodium stearate, presenting high respiration performances and no overt toxicity on lung cells, could be used to improve therapeutic outcomes of patient with Cystic Fibrosis (CF). PMID- 19184618 TI - Relative importance of intestinal and hepatic glucuronidation-impact on the prediction of drug clearance. AB - PURPOSE: To assess the extent of intestinal and hepatic glucuronidation in vitro and resulting implications on glucuronidation clearance prediction. METHODS: Alamethicin activated human intestinal (HIM) and hepatic (HLM) microsomes were used to obtain intrinsic glucuronidation clearance (CL(int,UGT)) for nine drugs using substrate depletion. The in vitro extent of glucuronidation (fm(UGT)) was determined using P450 and UGT cofactors. Utility of hepatic CL(int) for the prediction of in vivo clearance was assessed. RESULTS: fm(UGT) (8-100%) was comparable between HLM and HIM with the exception of troglitazone, where a nine fold difference was observed (8% and 74%, respectively). Scaled intestinal CL(int,UGT) (per g tissue) was six- and nine-fold higher than hepatic for raloxifene and troglitazone, respectively, and comparable to hepatic for naloxone. The remaining drugs had a higher hepatic than intestinal CL(int,UGT) (average five-fold). For all drugs with P450 clearance, hepatic CL(int,CYP) was higher than intestinal (average 15-fold). Hepatic CL(int,UGT) predicted on average 22% of observed in vivo CL(int); with the exception of raloxifene and troglitazone, where the prediction was only 3%. CONCLUSION: Intestinal glucuronidation should be incorporated into clearance prediction, especially for compounds metabolised by intestine specific UGTs. Alamethicin activated microsomes are useful for the assessment of intestinal glucuronidation and fm(UGT) in vitro. PMID- 19184619 TI - Self-rating of poor health: a comparison of Cuban elders in Havana and Miami. AB - This study examined the prevalence of self-reported 'poor health' among Cuban elders in Havana (n = 1,183) and Miami (n = 288) and explored the roles of chronic conditions and functional disability in their self-reports. A logistic regression model of reported poor health was estimated for each group, testing both independent and interactive effects of chronic conditions and functional disability. Despite differences in sociodemographic characteristics and disease/disability profiles between Cuban elders in the two cities, an equal proportion of each group (13%) reported poor health. Financial strain and functional disability were common predictors of self-reported poor health across the groups. Gender and chronic conditions were significant only in the Havana sample, among whom the likelihood of reporting poor health increased with female gender and the presence of chronic conditions. In the Havana sample, not only did chronic conditions have an independent effect, but also its interaction with functional disability was significant. The overall results suggest a particular role of chronic conditions in determining subjective health among elders in Havana. Findings are discussed in a sociocultural context. PMID- 19184620 TI - Intergenerational family relations and life satisfaction among three elderly population groups in transition in the Israeli multi-cultural society. AB - The study aims to illuminate the links between personal and familial resources and wellbeing of elders 65+ in three population groups in Israel: kibbutz members, new immigrants from the former Soviet Union and Arabs-all of whom are undergoing different types of personal, social and economic transitions. About 70 respondents in each group were interviewed regarding life satisfaction, familial relations based on the paradigm of intergenerational family solidarity and personal resources (socio-demographic and physical functioning). The main conclusions of this study are: the lives of the elderly immigrants are much more disruptive by the transitional migration processes they are undergoing and this affects their well-being which was much lower than the other two groups. Additionally they received more help from the family. Family solidarity, mainly opportunity structures and emotional bonds were especially strong among the Arabs, with the lowest level of conflict. The Arab elderly were also different from the other two groups in the lower level of help they provided to their adult children, probably due to their more limited level of personal resources and the differing social expectations. The majority of respondents acknowledged some degree of filial obligations, although much lower among kibbutz members. Personal resources (physical functioning and financial adequacy) had the strongest effect on life satisfaction in all three groups. The dimensions of family solidarity played a less dominant role. The discussion highlights the distinctive family culture of the three groups, the transition they face, and their differential resources with some policy recommendations. PMID- 19184621 TI - English use among older bilingual immigrants in linguistically concentrated neighborhoods: social proficiency and internal speech as intracultural variation. AB - This research focuses on patterns of English proficiency and use-of-English among older immigrants living in linguistically concentrated, ethnic neighborhoods. A sample (n = 60) of older Puerto Ricans, who moved from the island to the mainland in their twenties, were divided into English proficiency groups (fluent, high intermediate, low intermediate) via the Adult Language Assessment Scales. Participants then provided self-ratings of their English proficiency (understanding, speaking, reading, and writing), their use of English in social domains (language spoken with own-family, in-laws, spouse, children, neighbors, and workmates), and their use of English in private psychological domains (language of talking to oneself, counting, writing notes to oneself, thinking, dreaming, praying, and expressing feelings). Finally, all participants completed the Puerto Rican Bicultural Scale. Results show a cohort of immigrant elders whose first language is protected by their ethnic neighborhoods but whose domestic and private lives are increasingly permeated by English. In particular, children emerge as powerful forces of language socialization in English for their parents. Further, there are important individual differences by level of proficiency, with a lowest proficiency group that is less acculturated, lower in socioeconomic status, and even more linguistically isolated than groups with higher proficiency. In essence, level of second language proficiency is a potent source of intracultural variation. Methodologically, the paper makes the important point that self-rated patterns of language use are consistent with scores on formal measures of proficiency. The paper also provides empirical verification of the logic of dividing language use into external, social speech and internal, psychological speech. PMID- 19184623 TI - Pornographic actresses are a poor choice for assessing what men optimally prefer in women's looks: Comments on Voracek and Fisher (2006). PMID- 19184624 TI - Physicomechanical properties of naproxen-loaded microparticles prepared from Eudragit l100. AB - Microparticles of naproxen with Eudragit L100 and Aerosil were prepared by the emulsion solvent diffusion method in order to avoid local gastrointestinal irritation, one of the major side effects of nonsteroidal anti-inflammatory drugs after oral ingestion. The process of preparation involved the use of ethanol as good solvent, dichloromethane as a bridging liquid, water as poor solvent, Aerosil as anti-adhesion agent, and sodium dodecyl sulfate to aid in the dispersion of the drug and excipients into the poor solvent. The obtained microparticles were evaluated for micromeritic properties, yield, encapsulation efficiency, drug physical state, and drug release properties. The influence of formulation factors and preparation condition (polymer/naproxen ratio, Aerosil/polymer ratio, and the initial difference of temperature between the solvent and nonsolvent) on the properties of the microparticles were also examined. The resultant microparticles were finely spherical and uniform with high incorporation efficiency (>79%) and yield (>71%). The incorporation efficiency was enhanced with increasing the ratio of excipients to drug and the initial difference of temperature between the solvent and nonsolvent. The mean diameter of the microparticles was influenced by all of the manufacturing parameters. Studies carried out to characterize the micromeritic properties of formulations, such as flowability and packability, showed that microparticles were suitable for further pharmaceutical manipulation (e.g., capsule filling). Drug release studies of the microparticles confirmed the gastroresistance, and mathematical studies showed that the drug released followed a Hixon and Crowell kinetic. These microparticles represent a simple method for the preparation of drug-loaded enteric microparticles with desired micromeritic properties and gastroresistance release. PMID- 19184625 TI - SARS knowledge, perceptions, and behaviors: a comparison between Finns and the Dutch during the SARS outbreak in 2003. AB - BACKGROUND: The SARS outbreak served to test both local and international outbreak management and risk communication practices. PURPOSE: The study compares SARS knowledge, perceptions, behaviors, and information between Finns and the Dutch during the SARS outbreak in 2003. METHOD: The participants of the study, who used a modified SARS Psychosocial Research Consortium survey, were drawn from Internet panels in Finland (n = 308) and the Netherlands (n = 373) in June 2003. Multiple logistic regression analyses were used to calculate odds ratios (with 95% confidence intervals) to compare Finns with the Dutch for various levels of perceptions and behaviors. RESULTS: Adjusted for age, education, and income, Finns were more likely to be knowledgeable and worried about SARS as well as to have low perceived comparative SARS risk and poor personal efficacy beliefs about preventing SARS. Finns were also more likely than the Dutch to have high confidence in physicians on SARS issues but less likely to have received information from the Internet and have confidence in Internet information. CONCLUSIONS: The study shed light on how two European populations differed substantially regarding lay responses to SARS. Understanding these differences is needed in formulating and executing communication and outbreak management. PMID- 19184626 TI - Socioeconomic differences in lung cancer incidence: a systematic review and meta analysis. AB - OBJECTIVE: To investigate the associations between various socioeconomic indicators and lung cancer incidence. METHODS: We searched PubMed and EMBASE databases for studies on socioeconomic position (SEP) and lung cancer incidence published through October 2007. Random-effect model was used to pool the risk estimates from the individual studies. We stratified the analysis by adjustment strategy to investigate the influence of smoking on socioeconomic gradient in lung cancer incidence. RESULTS: Out of 3,288 citations, we identified 64 studies eligible for inclusion. Compared to the highest SEP level, we observed an overall increased risk in lung cancer incidence among people with low educational SEP (61%), low occupational SEP (48%), and low income-based SEP (37%). The negative social gradient for lung cancer incidence remained for most of the possible sets of pooled estimates obtained in subgroup analyses for occupational and educational SEP with less consistency for SEP based on income in studies adjusted and unadjusted for smoking. No evidence of publication bias was apparent. CONCLUSION: Lung cancer incidence was associated with low educational, occupational, and income-based SEP. The association, adjusted or unadjusted for smoking, points out the importance of social position to be addressed in all discussions on cancer preventive measures. PMID- 19184627 TI - Analysis of iodide and iodate in Lake Mead, Nevada using a headspace derivatization gas chromatography-mass spectrometry. AB - We report here a derivatization headspace method for the analysis of inorganic iodine in water. Samples from Lake Mead, the Las Vegas Wash, and from Las Vegas tap water were examined. Lake Mead and the Las Vegas Wash contained a mixture of both iodide and iodate. The average concentration of total inorganic iodine (TII) for Lake Mead was approximately 90 nM with an iodide-to-iodate ratio of approximately 1. The TII concentration (approximately 160 nM) and the ratio of iodide to iodate were higher for the Las Vegas Wash (approximately 2). The TII concentration for tap water was close to that of Lake Mead (approximately 90 nM); however, tap water contained no detectable iodide as a result of ozonation and chlorine treatment which converts all of the iodide to iodate. PMID- 19184628 TI - Effects of chemically spiked sediments on estuarine benthic communities: a controlled mesocosm study. AB - Ambient sediments were collected from a reference site in the North Edisto River, SC and transferred to a laboratory facility to investigate effects of chemical contaminants on estuarine infaunal communities under controlled mesocosm conditions. Sediment contaminant slurries were prepared using dried sediments collected from the reference site and spiked with a metal (copper), a polycyclic aromatic hydrocarbon (pyrene), and a pesticide (4,4(') dichlorodiphenyltrichloroethane) to yield nominal mean effects range-median (ERM) quotients of <0.01 (no addition), 0.1, and 1.0 and applied to control, low dose (TRT A), and high dose (TRT B) treatment groups, respectively. Sediment samples for contaminant and benthic analyses were collected at the start of the experiment, 1 month after dosing, and 3 months after dosing. Near-nominal mean ERM quotients of 0.001, 0.075, and 0.818 were measured initially after dosing and remained fairly constant throughout the experiment. Measures of benthic condition, diversity, and richness were significantly reduced in both treatment groups relative to the control 1 month after dosing and persisted in TRT B at 3 months. The results demonstrate that benthic community effects can be observed at mean ERM quotients that are about an order of magnitude lower than levels that have been shown to be associated with significant toxicity in acute laboratory bioassays with single species (e.g., amphipods) in other studies. PMID- 19184629 TI - Assessment of nitrate concentration in groundwater in Saudi Arabia. AB - Contamination of groundwater by nitrate is considered a global problem. Nitrates are introduced in the groundwater from a variety of sources like agricultural activities, poor sewer system, wastewaters, and industrial activities. In the present research, a survey of wells (n = 1,060) was undertaken in all 13 regions of the Kingdom of Saudi Arabia to assess the contained nitrate (NO(3)) levels. The results indicated variation in nitrate levels from 1.1 to 884.0 mg/L as NO(3) throughout the Kingdom. The average nitrate levels in milligrams per liter as NO(3) were as follows in descending order: 65.7 (Jizan), 60.3 (Asir), 60.0 (Qassim), 51.3 (Hail), 41.8 (Makkah Al Mukaramma), 41.3 (Madina Al Munnawara), 38.0 (Al Baha), 37.0 (Najran), 30.7, (Tabouk), 25.2 (Eastern Province), 18.8 (Riyadh), 15.8 (Al Jouf), and 9.1 (Hadwed Shamalyah). The results indicated that nitrate levels exceeded the maximum contaminant limits for drinking water (45 mg/L as NO(3)) in a number of wells (n = 213) in different regions of the Kingdom. The maximum and minimum wells exceeding the maximum contaminant limits for nitrate in drinking water were in Jizan (52.6%) and Hadwed Shamalyah (4.9%), respectively. Most of the wells which exceeded the maximum allowed limits for nitrate were in the areas which were used for agricultural and residential purposes. PMID- 19184630 TI - Three-class classification models of logS and logP derived by using GA-CG-SVM approach. AB - In this investigation, three-class classification models of aqueous solubility (logS) and lipophilicity (logP) have been developed by using a support vector machine (SVM) method combined with a genetic algorithm (GA) for feature selection and a conjugate gradient method (CG) for parameter optimization. A 5-fold cross validation and an independent test set method were used to evaluate the SVM classification models. For logS, the overall prediction accuracy is 87.1% for training set and 90.0% for test set. For logP, the overall prediction accuracy is 81.0% for training set and 82.0% for test set. In general, for both logS and logP, the prediction accuracies of three-class models are slightly lower by several percent than those of two-class models. A comparison between the performance of GA-CG-SVM models and that of GA-SVM models shows that the SVM parameter optimization has a significant impact on the quality of SVM classification model. PMID- 19184631 TI - Nucleocapsid of rabies virus improve immune response of an inactivated avian influenza vaccine. AB - The purpose of this study was to determine if nucleocapsid of rabies virus could improve the immune response (humoral and protective) of chickens vaccinated against avian influenza with an inactivated avian influenza experimental vaccine (AIV). On the other hand, AIV with and without NC was compared with an inactivated oil emulsion avian influenza commercial vaccine (CV) virus, currently used in Mexico. Groups of 8 day old chickens were vaccinated intracutaneously with an AIV (group 1); group 2, AIV supplemented with 20 microg of nucleocapsid of rabies virus (NC); Group 3, commercial vaccine (CV) and control groups (4 and 5) with 20 microg of NC and non-infected allantoic fluid, respectively. CV showed a better antibody-mediated response (p < 0.001) after and before challenging; which correlated with the best protection; while NC improved the protection in comparison with group 1. This is the first report on the potential utility of the rabies virus N protein to improve immune response in domestic species. PMID- 19184632 TI - Distribution of eosinophil granulocytes and mast cells in the reproductive tract of female goats in the preimplantation phase. AB - Changes in eosinophil granulocytes and mast cells post-insemination may affect conceptus implantation, but information regarding the numbers of such cells in the mammalian reproductive tract is limited. This study investigated the preimplantation distribution of eosinophil granulocytes and mast cells (MCs) in the reproductive tract organs of female goats. Uterus, uterine cervix and uterine tubes samples were obtained at slaughter. Cornu uteri were washed in phosphate buffer solution (each animal contained at least one embryo). Tissues were fixed in 10% neutral buffered formol, Carnoy solution and Mota's fixative (basic lead acetate) for 48 h and embedded in paraffin. Six-micrometre-thick sections were stained with Congo red (for eosinophil granulocytes) and toluidine blue in 1% aqueous solution at pH 1.0 for 5 min (for MCs). In the uterus, MCs occurred in highest numbers in the myometrium. Higher MC numbers were observed in uterus, uterine and uterine tubes in the preimplantation (experimental) group (cycle synchronised through 7 days intravaginal sponge with 0.3 g P(4)) compared with the control group (P < 0.05). Eosinophil granulocyte numbers were significantly higher in the experimental group than in the control group (P < 0.05). These results indicate preimplantation-related changes in numbers of eosinophil granulocytes and MCs in goat reproductive tract organs. PMID- 19184633 TI - Correlation between quantitative angiographic and intravascular ultrasound parameters in patients treated with sirolimus analogous-eluting stents. AB - Despite the undeniable contribution of intravascular ultrasound (IVUS) and quantitative coronary angiography (QCA) to assess drug-eluting stent (DES) effectiveness, the way these image modalities correlate to each other and to target-lesion revascularization (TLR) after PCI, is yet to be established. Thus we sought to evaluate whether there is an acceptable correlation between QCA and IVUS after DES implantation. We analyzed 204 pts treated with DES: Zotarolimus- (126), Sirolimus- (57), and Biolimus (31) with baseline and follow-up QCA and IVUS. The correlation between QCA lumen loss (LL) and intimal hyperplasia (IH) volume obstruction by IVUS was assessed by multiple regression analysis. Two QCA parameters (in-segment diameter stenosis and in-segment LL) and one IVUS variable (in-stent volume of IH) were evaluated as quantitative surrogates of 6 month TLR. The receiver operating characteristic method with c-statistics was used to assess the ability of each surrogate endpoint to predict TLR. QCA LL correlated positively with IVUS IH volume of obstruction (r = 0.69; CI95% 0.61-0.75: P < 0.0001), independent of DES type. The 2 QCA parameters were superior to the IVUS parameter as surrogates for TLR. Of note, QCA LL (c = 0.99) correlated best with TLR, even better than percent DS. In the DES era there is a good correlation between QCA measured LL and IVUS IH volume and therefore can be used as a surrogate of DES efficacy. PMID- 19184634 TI - Effects of resistin expression on glucose metabolism and hepatic insulin resistance. AB - In order to observe the effect of increased serum resistin on glucose metabolism, insulin sensitivity, and hepatic insulin resistance (IR), mice were intravenously injected with recombinant adenovirus carrying the resistin gene (Adv-resistin EGFP). Changes in hepatic glucose metabolism were observed using the Periodic Acid-Schiff method. Hepatic AMP-activated protein kinase (AMPK) activation was assessed by Western blot analysis, and glucose-6-phosphatase (G6Pase) and phosphoenolpyruvate carboxykinase (PEPCK) mRNA expression was determined using real-time RT-PCR. Although no effect on fasting blood glucose was detected, increased fasting insulin levels, decreased glucose tolerance and insulin sensitivity, and reduced hepatic glycogen levels and AMPK activation were seen in the Adv-resistin-EGFP mice. Finally, elevated G6Pase and PEPCK mRNA expression levels were detected upon overexpression of resistin. Resistin may inhibit hepatic AMPK activity, which results in elevated expression of gluconeogenic enzymes thereby affecting glucose metabolism and leading to decreased glycogen storage that contributes to the development of hepatic IR. PMID- 19184635 TI - Separation of SSEA-4 and TRA-1-60 labelled undifferentiated human embryonic stem cells from a heterogeneous cell population using magnetic-activated cell sorting (MACS) and fluorescence-activated cell sorting (FACS). AB - A major concern in human embryonic stem cell (hESC)-derived cell replacement therapy is the risk of tumorigenesis from undifferentiated hESCs residing in the population of hESC-derived cells. Separation of these undifferentiated hESCs from the differentiated derivatives using cell sorting methods may be a plausible approach in overcoming this problem. We therefore explored magnetic activated cell sorting (MACS) and fluorescence activated cell sorting (FACS) to separate labelled undifferentiated hESCs from a heterogeneous population of hESCs and hepatocellular carcinoma cells (HepG2) deliberately mixed respectively at different ratios (10:90, 20:80, 30:70, 40:60 and 50:50) to mimic a standard in vitro differentiation protocol, instead of using a hESC-differentiated cell population, so that we could be sure of the actual number of cells separated. HES 3 and HES-4 cells were labelled in separate experiments for the stem cell markers SSEA-4 and TRA-1-60 using primary antibodies. Anti-PE magnetic microbeads that recognize the PE-conjugated SSEA-4 labelled hESCs was added to the heterogeneous cell mixture and passed through the MACS column. The cells that passed through the column ('flow-through' fraction) and those retained ('labelled' fraction') were subsequently analysed using FACS. The maximum efficacy of hESCs retention using MACS was 81.0 +/- 2.9% (HES-3) and 83.6 +/- 4.2% (HES-4). Using FACS, all the undifferentiated hESCs labelled with the two cell-surface markers could be removed by selective gating. Both hESCs and HepG2 cells in the 'flow-through' fraction following MACS separation were viable in culture whereas by FACS separation only the HepG2 cells were viable. FACS efficiently helps to eliminate the undifferentiated hESCs based on their cell-surface antigens expressed. PMID- 19184637 TI - Cirrhosis and trauma are a lethal combination. AB - BACKGROUND: The objective of this study was to evaluate the risk of mortality and complications associated with cirrhosis in trauma patients. METHODS: This is an IRB-approved retrospective trauma registry study of patients admitted to an academic level 1 trauma center from 1997 to 2006. The following parameters were abstracted for analysis: age, gender, mechanism of injury, Abbreviated Injury Score, Injury Severity Score, Glasgow Coma Scale, mortality, and complications (ARDS, acute renal failure, pneumonia, intra-abdominal abscess, trauma-associated coagulopathy). Multivariable analysis was utilized to compare the mortality and complication rates between cirrhotic and noncirrhotic trauma patients. The subgroup of patients who underwent laparotomy was also analyzed. RESULTS: During the 10-year study period there were 36,038 trauma registry patients, of which 468 (1.3%) had a diagnosis of cirrhosis. The mortality in the cirrhotic group was 12% vs. 6% in the noncirrhotic group [adjusted odds ratio = 5.65 (95% CI = 3.72 - 8.41, p < 0.0001)]. ARDS, trauma-associated coagulopathy, and septic complications were significantly more common in the cirrhotic group. The overall severe complication rate in the two groups was 10 and 4%, respectively [adjusted odds ratio = 2.05 (95% CI = 1.45 - 2.84, p < 0.0001)]. For the subgroup of patients who underwent emergent abdominal exploration, the mortality rate increased to 40% compared with that of noncirrhotics at 15% [adjusted odds ratio = 4.35 (95% CI = 2.00 - 9.18, p = 0.0002)]. CONCLUSION: Cirrhosis is an independent risk factor for increased mortality and higher complication rate following trauma. Injured patients who undergo laparotomy are significantly more likely to die than noncirrhotic patients. Injured patients with cirrhosis warrant aggressive monitoring and treatment. PMID- 19184636 TI - Familial parathyroid tumors: diagnosis and management. AB - BACKGROUND: The management of hyperparathyroidism (HPT) in the familial setting is complex. Due to the rarity of familial HPT and its different presentation within and between the familial syndromes and individual kindreds, treatment recommendations based on high levels of evidence cannot be made. However, based on the molecular genetic studies and case series from institutions with significant experience, important management principles (grade C recommendations) have been developed. METHODS: We conducted a systematic review of the literature using evidence-based criteria. RESULTS: Issue 1: initial operation in multiple endocrine neoplasia type 1 (MEN1), a grade C recommendation can be made for subtotal parathyroidectomy. Issue 2: initial surgery in MEN 2A, a grade C recommendation can be made for excision of enlarged glands only. Issue 3: surgery in familial isolated HPT and HPT-jaw tumor (HPT-JT) syndrome may be treated with parathyroidectomy that is subtotal or less, although the risk of parathyroid cancer in HPT-JT requires attention (no grade of recommendation). Issue 4: parathyroid surgery in familial HPT syndromes in the setting of underlying mutations in the calcium receptor (CASR) gene involves subtotal parathyroidectomy (no grade of recommendation). Issue 5: the use of intraoperative PTH measurements in familial HPT may guide the extent of parathyroid resection (no grade of recommendation). CONCLUSIONS: The goals of parathyroidectomy in familial HPT are to achieve and maintain normocalcemia for the longest time possible, avoid both iatrogenic hypocalcemia and operative complications, and facilitate future surgery for recurrent disease. PMID- 19184638 TI - Ethics of surgical complications. PMID- 19184639 TI - Open surgical is superior to percutaneous access for radiofrequency ablation of hepatic metastases. AB - PURPOSE: This study was designed to determine the best approach to radiofrequency ablation (RFA) in the liver. METHODS: From a total of 41 procedures, 37 patients with 47 tumors were treated with RFA for metastatic disease. Indications included colorectal cancer (n=28, 68%), neuroendocrine tumors (n=2, 5%), gynecological primaries (n=4, 10%), pancreatic/duodenal cancer (n=2, 5%), and miscellaneous entities (n=5, 12%). Mean follow-up period was 18 (median, 18) months. All ways of approach to RFA were applied: percutaneous was chosen in 17 (41.5%), laparoscopic and hand-assisted laparoscopic in 5 (12.2%), and open surgical in 19 cases (46.3%), and in 10 cases, RFA was combined with hepatic resection. The average maximum tumor size was 2.3 (range, 0.8-6) cm, and the mean number of nodules treated per patient in a single session was 1.3 (range, 1-3). RESULTS: Overall survival was 59.5% at 2 years, recurrence-free 2-year survival was 12.6%, local tumor recurrence rate was 34%, and overall recurrence was 75.6%. Local tumor recurrence and disease-free survival were significantly improved in the open surgically treated patients compared with the percutaneous treatment group (15.8% [n=3] vs. 58.8% [n=10] and 11.5 vs. 7.9 months, p<0.01 [chi2 test] and p<0.05 [log-rank test], respectively). CONCLUSIONS: Open surgical approach is superior to percutaneous access for RFA in metastatic hepatic disease. PMID- 19184640 TI - A single residue in the 126-kDa protein of pepper mild mottle virus controls the severity of symptoms on infected green bell pepper plants. AB - Infectious cDNA clones originally derived from a mild strain of Pepper mild mottle virus were constructed by replacing residue 649, a critical point for attenuation of this virus, with all possible amino acids. All clones were infectious to pepper plants and induced a variety of symptoms, including no visible symptoms. The results of this study showed that a single amino acid mutation at residue 649 could control the function of the 126- and 183-kDa proteins, replicases with multiple roles in the life cycle of this virus. PMID- 19184641 TI - Quality of radiosurgery for single brain metastases with respect to treatment technology: a matched-pair analysis. AB - OBJECTIVE: A comparison of treatment parameters and quality of clinical outcome in patients with single brain metastases treated with two different technologies for radiosurgery (Gamma Knife and CyberKnife). METHODS: Treatment parameters were statistically analyzed. Clinical outcome was assessed by matched-pair analysis based on the treatment device, differences in dose prescription, and the date of the treatment. Patients were matched according to their tumor size, age, gender, primary cancer, and Radiation Therapy Oncology Group score. Survival post radiosurgery, local and distant tumor control, and complications were analyzed. Predictive factors were investigated. RESULTS: 423 single brain metastases were treated with Gamma Knife and 73 with CyberKnife. Tumor volumes were similar. The parameters minimum tumor dose, maximum tumor dose, prescription isodose volume, conformality index, homogeneity index, volume of tissue receiving a dose of 10 Gy or more were significantly larger in Gamma Knife group. Sixty-three patients were good matches. These showed the same pattern in parameters. Concerning the outcome analysis, only overall survival differed significantly between groups, twice as long with CyberKnife (P < 0.03). According to pooled data, dose was predictive of local failure, whole brain radiation therapy and chemotherapy were predictive of toxicity, the Radiation Therapy Oncology Group score was predictive of survival after radiosurgery, and date of treatment was predictive of overall survival. No factor predicted new brain metastases, including whole brain radiation therapy. CONCLUSIONS: The most important result of this study was the finding that the obvious differences in treatment-related parameters between Gamma Knife and CyberKnife had no impact on the quality of the clinical outcome after radiosurgery. Survival time increased chronologically, presumably due to an intensified anti-cancer therapy in the more recent era of the CyberKnife treatments. PMID- 19184642 TI - Multiple intracranial melanoma metastases: case report and review of the literature. AB - Although intracerebral metastases of malignant melanoma are common, those located in the sellar region and within the pontocerebellar area are extremely rare. Furthermore, to our knowledge, there is no report about melanoma metastasis to the epiphysis published so far. We report here a 46-year-old patient who had metastatic lesions in the sellar region, cerebellopontine area and epiphysial gland, preceded by a primary melanoma at her left shoulder. The diagnosis of sellar metastasis was confirmed histopathologically following a stereotactic biopsy. The patient received whole-brain irradiation therapy combined with chemotherapy. After 10 months, she died from a severe hemorrhage in the cerebellopontine angle. Autopsy findings confirmed melanoma metastases both in the cerebellopontine angle and additionally in the epiphysial gland. To our knowledge, this is the first case of multiple intracranial melanoma metastases including the suprasellar region, the pontocerebellar and epiphysial area. PMID- 19184643 TI - Molecular evolution of PKD2 gene family in mammals. AB - PKD2 gene encodes a critical cation channel protein that plays important roles in various developmental processes and is usually evolutionarily conserved. In the present study, we analyzed the evolutionary patterns of PKD2 and its homologous genes (PKD2L1, PKD2L2) from nine mammalian species. In this study, we demonstrated the orthologs of PKD2 gene family evolved under a dominant purifying selection force. Our results in combination with the reported evidences from functional researches suggested the entire PKD2 gene family are conserved and perform essential biological roles during mammalian evolution. In rodents, PKD2 gene family members appeared to have evolved more rapidly than other mammalian lineages, probably resulting from relaxation of purifying selection. However, positive selection imposed on synonymous sites also potentially contributed to this case. For the paralogs, our results implied that PKD2L2 genes evolved under a weaker purifying selection constraint than PKD2 and PKD2L1 genes. Interestingly, some loop regions of transmembrane domain of PKD2L2 exhibited higher P (N)/P (S) ratios than expected, suggesting these regions are more functional divergent in organisms and worthy of special attention. PMID- 19184644 TI - Global patterns of nicotine and tobacco consumption. AB - Humans consume tobacco in dozens of guises, all of which are toxic; globally, a tenth of deaths among adults are caused by tobacco. Tobacco may be combusted (e.g., cigarettes, bidis, kreteks); heated (e.g., waterpipes, hookah, nargile); or taken orally or nasally (e.g., snuff, betel quid, chewing tobacco). The predominant forms vary among cultures, but the use of cigarettes has grown most dramatically in the past century. While smoking rates among women are comparable to those among men in Europe and North America, in other regions the rate is ten or more times higher among men; this gender gap is closing among young people. Per capita tobacco use in the USA doubled in the first half of the twentieth century, and has since declined to less than the 1900 levels. While cigarettes were only 2% of tobacco consumed in the USA in 1900 (half was chewing tobacco) 50 years later they were over 80%. A similar increase in tobacco consumption, and a shift to cigarettes, has been occurring globally, with a concomitant increase in tobacco-related death and disease that is not expected to peak for another two decades. PMID- 19184646 TI - Nicotine content and delivery across tobacco products. AB - Nicotine is the principal alkaloid in both commercial and homemade products (e.g., cigarettes, smokeless tobacco, bidis, waterpipes) followed by nornicotine, anabasine, anatabine, and many other basic substances that contain a cyclic nitrogenous nucleus. Tobacco types, leaf position on the plant, agricultural practices, fertilizer treatment, and degree of ripening are among some prominent factors that determine the levels of alkaloids in tobacco leaf. From a random examination of 152 cultivated varieties of Nicotiana tabacum, a range of alkaloid variation between 0.17 and 4.93% was determined. In fact, every step in tobacco production that affects plant metabolism will influence the level of alkaloid content to a certain degree. Depending on blending recipe, type and amount of additives, and product design, all types of tobacco products contain a very wide range of nicotine concentration. However, the ultimate emission of nicotine to the user, exposure, and psychophar-macological effects depend not only on the content and emission, but also on the relationship between the product and the user. PMID- 19184647 TI - The road to discovery of neuronal nicotinic cholinergic receptor subtypes. AB - The discovery that mammalian brain expresses the mRNAs for nine different nicotinic cholinergic receptor subunits (alpha2-alpha7, beta2-beta4) that form functional receptors when expressed in Xenopus laevis oocytes suggests that many different types of nicotinic cholinergic receptors (nAChRs) might be expressed in the mammalian brain., Using an historical approach, this chapter reviews some of the progress made in identifying the nAChR subtypes that seem to play a vital role in modulating dopaminergic function. nAChR subtypes that are expressed in dopamine neurons, as well as neurons that interact with dopamine neurons (glutamatergic, GABAergic), serve as the focus of this review. Subjects that are highlighted include the discovery of a low affinity alpha4beta2* nAChR, the identity of recently characterized alpha6* nAChRs, and the finding that these alpha6* receptors have the highest affinity for receptor activation of any of the native receptors that have been characterized to date. Topics that have been ignored in other recent reviews of this area, such as the discovery and potential importance of alternative transcripts, are presented along with a discussion of their potential importance. PMID- 19184645 TI - Nicotine chemistry, metabolism, kinetics and biomarkers. AB - Nicotine underlies tobacco addiction, influences tobacco use patterns, and is used as a pharmacological aid to smoking cessation. The absorption, distribution and disposition characteristics of nicotine from tobacco and medicinal products are reviewed. Nicotine is metabolized primarily by the liver enzymes CYP2A6, UDPglucuronosyltransferase (UGT), and flavin-containing monooxygenase (FMO). In addition to genetic factors, nicotine metabolism is influenced by diet and meals, age, sex, use of estrogen-containing hormone preparations, pregnancy and kidney disease, other medications, and smoking itself. Substantial racial/ethnic differences are observed in nicotine metabolism, which are likely influenced by both genetic and environmental factors. The most widely used biomarker of nicotine intake is cotinine, which may be measured in blood, urine, saliva, hair, or nails. The current optimal plasma cotinine cut-point to distinguish smokers from non-smokers in the general US population is 3 ng ml(-1). This cut-point is much lower than that established 20 years ago, reflecting less secondhand smoke exposure due to clear air policies and more light or occasional smoking. PMID- 19184648 TI - Magnetic resonance imaging studies of cigarette smoking. AB - This chapter reviews studies that have applied magnetic resonance imaging (MRI) toward a better understanding of the neurobiological correlates and consequences of cigarette smoking and nicotine dependence. The findings demonstrate that smokers differ from nonsmokers in regional brain structure and neurochemistry, as well as in activation in response to smoking-related stimuli and during the execution of cognitive tasks. We also review functional neuroimaging studies on the effects of nicotine administration on brain activity, both at rest and during the execution of cognitive tasks, independent of issues related to nicotine withdrawal and craving. Although chronic cigarette smoking is associated with poor cognitive performance, acute nicotine administration appears to enhance cognitive performance and increase neural efficiency in smokers. PMID- 19184650 TI - Molecular and cellular mechanisms of action of nicotine in the CNS. AB - Nicotine achieves its psychopharmacological effects by interacting with nicotinic acetylcholine receptors (nAChRs) in the brain. There are numerous subtypes of nAChR that differ in their properties, including their sensitivity to nicotine, permeability to calcium and propensity to desensitise. The nAChRs are differentially localised to different brain regions and are found on presynaptic terminals as well as in somatodendritic regions of neurones. Through their permeability to cations, these ion channel proteins can influence both neuronal excitability and cell signalling mechanisms, and these various responses can contribute to the development or maintenance of dependence. However, many questions and uncertainties remain in our understanding of these events and their relevance to tobacco addiction. In this chapter, we briefly overview the fundamental characteristics of nAChRs that are germane to nicotine's effects and then consider the cellular responses to acute and chronic nicotine, with particular emphasis on dopamine systems because they have been the most widely studied in the context of nicotine dependence. Where appropriate, methodological aspects are critically reviewed. PMID- 19184651 TI - The neuronal pathways mediating the behavioral and addictive properties of nicotine. AB - This chapter considers the neurobiological mechanisms that are thought to mediate the reinforcing or rewarding properties of nicotine. It focuses on the data (derived principally from studies with experimental animals) showing that nicotine, like other drugs of dependence, stimulates the mesolimbic dopamine (DA) neurones that project to the nucleus accumbens and that these effects play a pivotal role in the biology underlying nicotine dependence. The reinforcing or rewarding properties of nicotine are thought to be associated particularly with the increase in DA overflow evoked in the shell subdivision of the accumbens. However, behavioural studies suggest that these properties of nicotine in experimental animals do not seem to be sufficiently potent to explain the powerful addiction to tobacco experienced by most habitual smokers. This chapter also considers the biological mechanisms that mediate the effects of cues and stimuli associated with the presentation of nicotine, which are thought to contribute significantly to the powerful addictive properties of tobacco smoke. PMID- 19184649 TI - In vivo brain imaging of human exposure to nicotine and tobacco. AB - While most cigarette smokers endorse a desire to quit smoking, only 14-49% will achieve abstinence after 6 months or more of treatment. A greater understanding of the effects of smoking on brain function may result in improved pharmacological and behavioral interventions for this condition. Research groups have examined the effects of acute and chronic nicotine/cigarette exposure on brain activity using functional imaging; the purpose of this chapter is to synthesize findings from such studies and present a coherent model of brain function in smokers. Responses to acute administration of nicotine/smoking include reduced global brain activity; activation of the prefrontal cortex, thalamus, and visual system; activation of the thalamus and visual cortex during visual cognitive tasks; and increased dopamine (DA) concentration in the ventral striatum/nucleus accumbens. Responses to chronic nicotine/cigarette exposure include decreased monoamine oxidase (MAO) A and B activity in the basal ganglia and a reduction in alpha4beta2 nicotinic acetylcholine receptor (nAChR) availability in the thalamus and putamen (accompanied by an overall upregulation of these receptors). These findings indicate that smoking enhances neurotransmission through cortico-basal ganglia-thalamic circuits by direct stimulation of nAChRs, indirect stimulation via DA release or MAO inhibition, or a combination of these and possibly other factors. Activation of this circuitry may be responsible for the effects of smoking seen in tobacco-dependent smokers, such as improvements in attentional performance, mood, anxiety, and irritability. PMID- 19184652 TI - Molecular genetics of nicotine metabolism. AB - The molecular genetics of nicotine metabolism involves multiple polymorphic catalytic enzymes. Variation in metabolic pathways results in nicotine disposition kinetics that differ between individuals and ethnic groups. Twin studies indicate that a large part of this variance is genetic in origin, although environmental influences also contribute. The primary aim of this chapter is to review the current knowledge regarding the genetic variability in the enzymes that metabolize nicotine in humans. The focus is on describing the genetic polymorphisms that exist in cytochromes P450 (CYPs), aldehyde oxidase 1 (AOX1), UDP-glucuronosyltransferases (UGTs), and flavin-containing monooxygenase 3 (FMO3). Genetic studies have demonstrated that polymorphisms in CYP2A6, the primary enzyme responsible for nicotine breakdown, make a sizable contribution to the wide range of nicotine metabolic capacity observed in humans. Thus, special attention will be given to CYP2A6, because slower nicotine metabolism requires less frequent self-administration, and accordingly influences smoking behaviors. In addition, the molecular genetics of nicotine metabolism in nonhuman primates, mice, and rats will be reviewed briefly. PMID- 19184653 TI - Sex differences in nicotine action. AB - Accumulating evidence suggests that the antecedents, consequences, and mechanisms of drug abuse and dependence are not identical in males and females and that gender may be an important variable in treatment and prevention. Although there has been a decline in smoking prevalence in developed countries, females are less successful in quitting. Tobacco use is accepted to be a form of addiction, which manifests sex differences. There is also evidence for sex differences in the central effects of nicotine in laboratory animals. Although social factors impact smoking substantially in humans, findings from nonhuman subjects in controlled experiments provide support that sex differences in nicotine/tobacco addiction have a biological basis. Differences in the pharmacokinetic properties of nicotine or the effect of gonadal hormones may underlie some but not all sex differences observed. Laboratory-based information is very important in developing treatment strategies. Literature findings suggest that including sex as a factor in nicotine/tobacco-related studies will improve our success rates in individually tailored smoking cessation programs. PMID- 19184654 TI - Recognising nicotine: the neurobiological basis of nicotine discrimination. AB - Drug discrimination methodology makes possible the objective, quantitative study of the perception of psychoactive drug effects in either human or animal subjects. Investigations of the nicotine discriminative stimulus complex have contributed to our present understanding of nicotine psychopharmacology by defining the origin of its effects at specific subtypes of nicotinic receptor and the role of diverse neurotransmitter systems as mediating and modulating mechanisms. The evidence strongly supports central sites as the origins of the nicotine stimulus, and these are likely to be located in the mesocorticolimbic dopaminergic neurons; the medial prefrontal cortex is primarily involved, with the Nucleus accumbens and ventral tegmental area of secondary importance, while another element of the complex stimulus may arise in the dorsal hippocampus. Additionally, it appears that interactions of nicotine with the dopamine, serotonin, cannabinoid and probably glutamate systems all contribute to the final perceived stimulus. The resemblance between the nicotine discriminative stimulus and those of the psychomotor stimulant drugs amphetamine and cocaine contributes to defining the nature of the addictive properties of nicotine. It is particularly interesting that acute and chronic exposure to caffeine produce quantitative and qualitative changes in the characteristics of the nicotine stimulus. Interactions of nicotine with caffeine and cannabinoids strengthen proposals that the use of one substance serves as a "gateway" in sequential shifts of the target substance for drug-seeking behaviour, with profound implications for the human use of the substances concerned. Drug discrimination is also an important standard technique used in assessments of the abuse liability of novel psychoactive compounds, with relevance to attempts to develop novel nicotinic agonists for use as cognitive enhancers. PMID- 19184656 TI - Discriminative stimulus effects of nicotine in humans. AB - Behavioral discrimination procedures clearly demonstrate that nicotine elicits interoceptive stimulus effects in humans that are malleable by various pharmacological manipulations as well as by some behavioral manipulations. The parameters of nicotine discrimination and both chronic and acute factors that may alter discrimination behavior are addressed in this chapter, which emphasizes research by the author involving nicotine delivered by nasal spray. Human discrimination of nicotine is centrally mediated, as the central and peripheral nicotine antagonist mecamylamine blocks discrimination but the peripheral antagonist trimethaphan does not. The threshold dose for discrimination of nicotine via spray appears to be very low in smokers as well as nonsmokers. Because smoked tobacco delivers nicotine more rapidly than spray, the threshold dose of nicotine via smoking is probably even lower. In terms of individual differences, smokers may become tolerant to the discriminative stimulus effects of higher nicotine doses but not of low doses. Men may be more sensitive than women to nicotine's discriminative stimulus effects, consistent with other research suggesting that nicotine is more reinforcing in men than in women. Other potential individual differences in nicotine discrimination have not been clearly tested, but may include genetics, obesity, and dependence on other drugs. Acute environmental factors that alter nicotine discrimination include the specific training and testing conditions, pointing to the need for careful control over such conditions during research. Other factors, such as concurrent acute use of alcohol or caffeine, do not appear to alter nicotine discrimination, suggesting that changes in nicotine discrimination are not likely explanations for the association of smoking behavior with use of those drugs. Concurrent physical activity also does not appear to alter nicotine discrimination, indicating that results from studies of discrimination in subjects at quiet rest, the standard approach in this research, generalize well to discrimination in subjects engaged in various activities, as often occurs in the natural environment. Future research should more clearly examine the potential role of nicotine's discriminative stimulus effects in nicotine reinforcement and determine the generalizability of these findings to nicotine delivered by other means, particularly tobacco smoking. PMID- 19184657 TI - Rodent models of nicotine withdrawal syndrome. AB - Simple, rapid and inexpensive rodent models of nicotine physical dependence and withdrawal syndrome have proved useful for preliminary screening of smoking cessation treatments. They have led to an exponential increase of knowledge regarding the underlying neurobiological mechanisms of dependence and withdrawal syndrome. The human nicotine withdrawal syndrome in smoking cessation is variable and multidimensional, involving irritability, anxiety, depression, cognitive and attentional impairments, weight gain, sleep disturbances, and craving for nicotine. Aside from sleep disturbances, analogous phenomena have been seen in rodent models using different measures of withdrawal intensity. It appears likely that different withdrawal phenomena may involve some partially divergent mechanisms. For example, depression-like phenomena may involve alterations in mechanisms such as the mesolimbic dopamine pathway from the ventral tegmental area to the nucleus accumbens. Irritability and anxiety may involve alterations in endogenous opioid systems and other regions, such as the amygdala. This chapter reviews many additional anatomical, neurochemical, and developmental elements that impact nicotine physical dependence. PMID- 19184658 TI - Approaches, challenges, and experience in assessing free nicotine. AB - Delivery of nicotine in the most desirable form is critical in maintaining people's use of tobacco products. Interpretation of results by tobacco industry scientists, studies that measure free-base nicotine directly in tobacco smoke, and the variability of free-base nicotine in smokeless tobacco products all indicate that the form of nicotine delivered to the tobacco user, in addition to the total amount, is an important factor in whether people continue to use the product following their initial exposure. The physiological impact of nicotine varies with the fraction that is in the free-base form and this leads to continued exposure to other toxic tobacco contents and emissions. In addition to evaluating the total nicotine delivered to the user, measuring the fraction of nicotine in the free-base form is critical in understanding and controlling the influence of nicotine on tobacco use. PMID- 19184659 TI - Tobacco industry manipulation of nicotine dosing. AB - For more than a half century, tobacco manufacturers have conducted sophisticated internal research to evaluate nicotine delivery, and modified their products to ensure availability of nicotine to smokers and to optimize its effects. Tobacco has proven to be a particularly effective vehicle for nicotine, enabling manipulation of smoke chemistry and of mechanisms of delivery, and providing sensory cues that critically inform patterns of smoking behavior as well as reinforce the impact of nicotine. A range of physical and chemical product design changes provide precise control over the quantity, form, and perception of nicotine dose, and support compensatory behavior, which is driven by the smoker's addiction to nicotine. Cigarette manufacturers also enhance the physiological effects of nicotine through the introduction and use of compounds that interact with nicotine but do not directly alter its form or delivery. A review of internal documents indicates important historical differences, as well as significant differences between commercial brands, underscoring the effectiveness of methods adopted by manufacturers to control nicotine dosing and target the needs of specific populations of smokers through commercial product development. Although the focus of the current review is on the manipulation of nicotine dosing characteristics, the evidence indicates that product design facilitates tobacco addiction through diverse addiction-potentiating mechanisms. PMID- 19184655 TI - Effects of nicotine in experimental animals and humans: an update on addictive properties. AB - Tobacco use through cigarette smoking is the leading preventable cause of death in the developed world. Nicotine, a psychoactive component of tobacco, appears to play a major role in tobacco dependence, but the reinforcing effects of nicotine have often been difficult to demonstrate directly in controlled studies with laboratory animals or human subjects. Here we update our earlier review published in Psychopharmacology (Berl) in 2006 on findings obtained with various procedures developed to study dependence-related behavioral effects of nicotine in experimental animals and humans. Results obtained with drug self-administration, conditioned place preference, subjective reports of nicotine effects and nicotine discrimination indicate that nicotine can function as an effective reinforcer of drug-seeking and drug-taking behavior both in experimental animals and humans under appropriate conditions. Interruption of chronic nicotine exposure produces ratings of drug withdrawal and withdrawal symptoms that may contribute to relapse. Difficulties encountered in demonstrating reinforcing effects of nicotine under some conditions, relative to other drugs of abuse, may be due to weaker primary reinforcing effects of nicotine, to aversive effects produced by nicotine, or to a more critical contribution of environmental stimuli to the maintenance of drug-seeking and drug-taking behavior with nicotine than with other drugs of abuse. Several recent reports suggest that other chemical substances inhaled along with nicotine in tobacco smoke may play a role in sustaining smoking behavior. However, conflicting results have been obtained with mice and rats and these findings have not yet been validated in nonhuman primates or human subjects. Taken together, these findings suggest that nicotine acts as a typical drug of abuse in experimental animals and humans in appropriate situations. PMID- 19184660 TI - Pharmacotherapy for tobacco dependence. AB - Pharmacotherapy can provide effective treatment of tobacco dependence and withdrawal, and thereby facilitate efforts to achieve and sustain tobacco abstinence. Currently approved medications for smoking cessation are nicotine replacement medications (NRT), including nicotine patch, gum, lozenge, sublingual tablet, inhaler and nasal spray, the antidepressant bupropion, and the nicotinic partial agonist varenicline. This review discusses the pharmacological basis for the use of these medications, and the properties that might contribute to their efficacy, safety, and abuse liability. The review also discusses how pharmacological principles can be used to improve existing medications, as well as assist in the development of new medications. PMID- 19184661 TI - Nicotine psychopharmacology: policy and regulatory. AB - Powerful nerve agent, poison, addictive drug, or wonder medicine of the future? Nicotine has had a long and storied history in pharmacology, physiology, public health and, more recently, in regulatory policy initiatives in the United States and internationally. Psychopharmacology research on nicotine and tobacco came to particular prominence in the latter third of the twentieth century with exploration addressing the role of nicotine in tobacco use, the potential categorization of nicotine as an addictive drug, the pharmacological basis for treatment of tobacco addiction, and the perspective of policy developers seeking to reduce the toll of tobacco use. In fact, the 2005 ratification of the World Health Organization's first global health treaty, the Framework Convention on Tobacco Control, provides further impetus for extending the science foundation for tobacco disease control and policy efforts. Implementation of the treaty's provisions will control tobacco use and reduce the 500 million premature deaths projected to occur in the first half of the twenty-first century from tobacco use. Psychopharmacological research on nicotine and tobacco was important in the rationale and development of the treaty. The public health relevance of psychopharmacology research continues to grow with the realization of the potential of nicotine and related drugs to treat or prevent a diverse range of disorders (e.g., Alzheimer's disease, attention deficit hyperactivity disorder, and pain). Although comprehensive review of the research and implications is beyond the scope of this article, the more modest goal of providing insight into the theoretical, clinical, and policy importance of key psychopharmacology research laboratories over the past few decades is attempted. PMID- 19184662 TI - QTL, additive and epistatic effects for SCN resistance in PI 437654. AB - PI 437654 is a unique accession because of its resistance to nearly all HG types (races) of soybean cyst nematode (Heterodera glycines Ichinohe; SCN). Objectives of this study were to confirm and refine the locations and gene action associated with SCN resistance previously discovered in PI 437654, and to identify new QTLs that may have been missed because of low coverage with genetic markers used in previous studies. Using 205 F(7:9) RILs and 276 SSR and AFLP molecular markers covering 2,406.5 cM of 20 linkage groups (LGs), we confirmed and refined the locations of major SCN resistance QTLs on LG-A2, -B1, and -G previously identified in PI 437654 or other resistant sources. We found that these major QTLs have epistatic effects among them or with other loci for SCN resistance. We also detected some new QTLs with additive or epistatic effects for SCN resistance to different HG types (races) on all LGs except LGs-B2 and -D1b. The QTL on LG-G was associated with resistance to HG types 2.5.7, 1.2.5.7, 0, and 2.7 (races 1, 2, 3, and 5), and it contributed a large proportion of the additive effects. The QTL on LG-A2 was associated with resistance to HG types 2.5.7 and 0 (races 1 and 3). The QTL on LG-B1, associated with resistance to HG types 2.5.7, 0, 2.7 (races 1, 3, and 5), was the similar QTL found in PI 90763 and PI 404198B. In addition to QTL on LGs-A2, -B1 and -G, a novel additive QTL associated with SCN resistance to HG types 0, 2.7, and 1.3.5.6.7 (race 3, 5, and 14) was identified on LG-I flanked by Sat_299 and Sat_189. Several minor QTLs on LGs-C1, D1a, H, and K were also found to be associated with SCN resistance. Confirmation of the new resistance QTL is underway by evaluating another RIL population with a different genetic background. PMID- 19184663 TI - Application of XPS and ToF-SIMS for surface chemical analysis of DNA microarrays and their substrates. AB - The chemical composition of the functional surfaces of substrates used for microarrays is one of the important parameters that determine the quality of a microarray experiment. In addition to the commonly used contact angle measurements to determine the wettability of functionalized supports, X-ray photoelectron spectroscopy (XPS) and time-of-flight secondary ion mass spectrometry (ToF-SIMS) are more specific methods to elucidate details about the chemical surface constitution. XPS yields information about the atomic composition of the surface, whereas from ToF-SIMS, information on the molecular species on the surface can be concluded. Applied on printed DNA microarrays, both techniques provide impressive chemical images down to the micrometer scale and can be utilized for label-free spot detection and characterization. Detailed information about the chemical constitution of single spots of microarrays can be obtained by high-resolution XPS imaging. PMID- 19184664 TI - Clinical application of a handy intraoperative measurement device for lumbar segmental instability. AB - We describe the development of a new device that permits handy intraoperative measurement of lumbar segmental instability. The subjects comprised 80 patients with lumbar degenerative disease. Relationships between preoperative radiological assessments and extended distance as measured using our new device were investigated. Mean extended distance measured using the device was 3.7 +/- 1.9 mm. Correlation coefficients between angular motion and extended distance, and translational motion and extended distance were 0.76 and 0.66, respectively, revealing significant positive relationships between these values (p < 0.01 each). The correlation coefficient between the intervertebral endplate angle on the flexion film and extended distance was -0.78, showing a significant negative relationship (p < 0.01). In conclusion, the device for intraoperative measurement of lumbar segmental instability that we have developed appears to permit simple measurement of intervertebral instability and provides operators with valuable information for selecting operative methods of spinal fusion or instrumentation. PMID- 19184665 TI - Rituximab in life threatening antisynthetase syndrome. AB - This case study reports a patient with severe interstitial pneumonitis, mild polyarthritis and polymyositis, accompanied by the presence of anti-Jo-1 antibodies diagnosed as antisynthetase syndrome. The concurrence of anti-Jo-1 with anti-Ro/SSA antibodies leads to a more severe form of interstitial lung disease. This patient was referred to our hospital because of life threatening respiratory failure. He was refractory to glucocorticoids and cyclophosphamide, but was successfully treated with two sequential infusions of rituximab. Clinical condition improved very rapidly. Response to treatment was well correlated with a fall of levels of serum soluble IL2-receptor. A decrease in pulmonary disease activity was visualized on PET-scans before and after two sequential rituximab infusions. PMID- 19184666 TI - Assessment of beta-amyloid deposits in human brain: a study of the BrainNet Europe Consortium. AB - beta-Amyloid (A-beta) related pathology shows a range of lesions which differ both qualitatively and quantitatively. Pathologists, to date, mainly focused on the assessment of both of these aspects but attempts to correlate the findings with clinical phenotypes are not convincing. It has been recently proposed in the same way as iota and alpha synuclein related lesions, also A-beta related pathology may follow a temporal evolution, i.e. distinct phases, characterized by a step-wise involvement of different brain-regions. Twenty-six independent observers reached an 81% absolute agreement while assessing the phase of A-beta, i.e. phase 1 = deposition of A-beta exclusively in neocortex, phase 2 = additionally in allocortex, phase 3 = additionally in diencephalon, phase 4 = additionally in brainstem, and phase 5 = additionally in cerebellum. These high agreement rates were reached when at least six brain regions were evaluated. Likewise, a high agreement (93%) was reached while assessing the absence/presence of cerebral amyloid angiopathy (CAA) and the type of CAA (74%) while examining the six brain regions. Of note, most of observers failed to detect capillary CAA when it was only mild and focal and thus instead of type 1, type 2 CAA was diagnosed. In conclusion, a reliable assessment of A-beta phase and presence/absence of CAA was achieved by a total of 26 observers who examined a standardized set of blocks taken from only six anatomical regions, applying commercially available reagents and by assessing them as instructed. Thus, one may consider rating of A-beta-phases as a diagnostic tool while analyzing subjects with suspected Alzheimer's disease (AD). Because most of these blocks are currently routinely sampled by the majority of laboratories, assessment of the A-beta phase in AD is feasible even in large scale retrospective studies. PMID- 19184667 TI - STREGA: a 'How-To' guide for reporting genetic associations. PMID- 19184669 TI - Gestural communication of the gorilla (Gorilla gorilla): repertoire, intentionality and possible origins. AB - Social groups of gorillas were observed in three captive facilities and one African field site. Cases of potential gesture use, totalling 9,540, were filtered by strict criteria for intentionality, giving a corpus of 5,250 instances of intentional gesture use. This indicated a repertoire of 102 gesture types. Most repertoire differences between individuals and sites were explicable as a consequence of environmental affordances and sampling effects: overall gesture frequency was a good predictor of universality of occurrence. Only one gesture was idiosyncratic to a single individual, and was given only to humans. Indications of cultural learning were few, though not absent. Six gestures appeared to be traditions within single social groups, but overall concordance in repertoires was almost as high between as within social groups. No support was found for the ontogenetic ritualization hypothesis as the chief means of acquisition of gestures. Many gestures whose form ruled out such an origin, i.e. gestures derived from species-typical displays, were used as intentionally and almost as flexibly as gestures whose form was consistent with learning by ritualization. When using both classes of gesture, gorillas paid specific attention to the attentional state of their audience. Thus, it would be unwarranted to divide ape gestural repertoires into 'innate, species-typical, inflexible reactions' and 'individually learned, intentional, flexible communication'. We conclude that gorilla gestural communication is based on a species-typical repertoire, like those of most other mammalian species but very much larger. Gorilla gestures are not, however, inflexible signals but are employed for intentional communication to specific individuals. PMID- 19184672 TI - Molecular cloning and evolutionary analysis of hemoglobin alpha-chain genes in bats. AB - Bats are the only mammals with the capacity for powered flight. When flying, they need abundant energy and oxygen. According to previous works, the hemoglobin (Hb) oxygen loading function of bats is insensitive to variations in body temperature, although different bat species have different heat sensitivity. We cloned Hb alpha-chain sequences from eight bat species to investigate whether they have different characteristics. We found that Hb in the bat lineages is under purifying selection, which accords with the importance of its function in bats. Three turn regions in bat Hb, however, have distinct evolutionary rates compared with those of other mammals, and the codons in these regions have an accelerated rate of evolution. These codons are under divergent selection in bats. These changes in Hb may have occurred in response to the physiological requirements of the species concerned, as adaptations to different lifestyles. PMID- 19184673 TI - Choice of study phenotype in osteoporosis genetic research. AB - Osteoporosis is a common disease characterized by compromised bone strength predisposing a person to an increased risk of osteoporotic fracture (OF). Recently, extensive efforts have been made to identify candidate genes underlying osteoporosis by the use of surrogate phenotypes, such as bone mineral density (BMD) and bone geometry. Among them, BMD is a suitable choice if we aim to classify the role of biological pathways for bone strength and to understand the bone conditions in the development of osteoporosis. However, evidences show that the genetic correlation between BMD and OF is very limited. In this review, we are mainly concerned with an important issue, i.e., phenotype choice in osteoporosis genetic research. For clarity, we address this issue with several arguments, and comments are made on most representative literature. PMID- 19184674 TI - A subgeneric classification of the genus Vaccinium and the metamorphosis of V. section Bracteata Nakai: more terrestrial and less epiphytic in habit, more continental and less insular in distribution. AB - The taxonomic integrity of Vaccinium section Bracteata sensu Sleumer was assessed using a variety of numerical measures on a data matrix created from 46 OTUs scored for 65 descriptors. These analyses supported a much restricted ambit for section Bracteata and the concomitant resurrection of section Nesococcus and section Euepigynium, a more cosmopolitan interpretation for section Eococcus and section Pyxothamnus as well as a new taxon, Vaccinium section Baccula-nigra Kloet, sect. nov. to accommodate V. fragile Franch. and its conspecifics. A key to all the sections as well as a brief description for each section is also provided. PMID- 19184668 TI - Strengthening the reporting of genetic association studies (STREGA): an extension of the STROBE Statement. AB - Making sense of rapidly evolving evidence on genetic associations is crucial to making genuine advances in human genomics and the eventual integration of this information in the practice of medicine and public health. Assessment of the strengths and weaknesses of this evidence, and hence the ability to synthesize it, has been limited by inadequate reporting of results. The STrengthening the REporting of Genetic Association studies (STREGA) initiative builds on the Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) Statement and provides additions to 12 of the 22 items on the STROBE checklist. The additions concern population stratification, genotyping errors, modeling haplotype variation, Hardy-Weinberg equilibrium, replication, selection of participants, rationale for choice of genes and variants, treatment effects in studying quantitative traits, statistical methods, relatedness, reporting of descriptive and outcome data, and the volume of data issues that are important to consider in genetic association studies. The STREGA recommendations do not prescribe or dictate how a genetic association study should be designed but seek to enhance the transparency of its reporting, regardless of choices made during design, conduct, or analysis. PMID- 19184675 TI - Productive and reproductive performance of strategically supplemented free grazing prepartum Bunaji cows in the agropastoral farming system. AB - The effects of prepartum supplementary feeding on the productive and reproductive performance were investigated using grazing gestating Bunaji cows with an average initial body weight of 294.50 +/- 3.75 kg. Twenty cows were allocated to a completely randomized design, with five animals per treatment. The treatments were: A, range grazing (RG); B, RG + 100% corn bran (CB); C, RG + 60% CB + 40% palm kernel cake (PKC), and D, RG + 60% CB + 40% dried brewer's grains (DBG). The average daily gains (ADG) and body condition scores (BCS) of supplemented cows were significantly better than the non-supplemented cows. Postpartum weight loss was markedly reduced in supplemented cows compared to their non-supplemented counterparts. Mean milk offtake and mean milk yield per lactation were significantly lower in non-supplemented cows than the supplemented ones. Among the supplemented cows, ADG, BCS, mean milk offtake and milk yield per lactation were significantly better for cows on treatments C and D than those on treatment B. Though insignificant, longest lactation length (LL) and shortest calving interval were obtained for supplemented cows. Calf's birth weight was similar among the treatments. Milk yield was significantly influenced (R(2) = 0.8601) by cow's weight, BCS and LL. PMID- 19184676 TI - Analytical tools for the physicochemical profiling of drug candidates to predict absorption/distribution. AB - The measurement of physicochemical properties at an early phase of drug discovery and development is crucial to reduce attrition rates due to poor biopharmaceutical properties. Among these properties, ionization, lipophilicity, solubility and permeability are mandatory to predict the pharmacokinetic behavior of NCEs (new chemical entities). Due to the high number of NCEs, the analytical tools used to measure these properties are automated and progressively adapted to high-throughput technologies. The present review is dedicated to experimental methods applied in the early drug discovery process for the determination of solubility, ionization constants, lipophilicity and permeability of small molecules. The principles and experimental conditions of the different methods are described, and important enhancements in terms of throughput are highlighted. PMID- 19184678 TI - The role of industry in brand or generic drug recognition. PMID- 19184677 TI - Zonula occludens-1, occludin, and E-cadherin protein expression in biliary tract cancers. AB - The incidence of cholangiocarcinomas originating from intrahepatic and extrahepatic bile ducts, as well as of gallbladder carcinoma is increasing worldwide. The malignant transformation of biliary epithelia involves profound alterations of proteins in the intercellular junctions, among others zonula occludens-1 (ZO-1), occludin, and E-cadherin. Each plays important role in the maintenance of epithelial cell polarity and regulation of cell growth and differentiation. Our aim was to investigate ZO-1, occludin, and E-cadherin immunohistochemical reactions in tissue microarray blocks containing 57 normal and 62 neoplastic biliary tract samples. We demonstrated that the tight junction components ZO-1, occludin, and E-cadherin are downregulated in carcinomas arising from various compartments of the biliary tract (normal intrahepatic and extrahepatic bile ducts, gallbladder) as compared with their normal sites of origin. These results were confirmed by discriminant analysis yielding clear separation of the three normal sample groups from carcinomas in the corresponding locations. PMID- 19184679 TI - Attitude change following a homeless clinic experience. PMID- 19184680 TI - Dealing with loss in the journey of mentoring. PMID- 19184681 TI - Leadership in times of crisis. PMID- 19184682 TI - Reliability and validity testing of an evidence-based medicine OSCE station. AB - The six competencies of the Accreditation Council for Graduate Medical Education include the lifelong learning skills of evidence-based medicine (EBM)/information mastery. We developed and tested an Objective Structured Clinical Examination (OSCE) station that would measure these skills in family medicine residents. This EBM OSCE station is a 30-minute station within a regular OSCE exam. It uses an 8 point checklist and global measure and has good psychometric properties, including construct validity, interrater reliability (correlation=.96), and internal reliability (Cronbach's=.58). This tool is useful for training programs, since assessing EBM/information mastery is an important part of the evaluation of physician skills. PMID- 19184683 TI - Affect and the art of healing. PMID- 19184686 TI - Lesson in a pill box: teaching about the challenges of medication adherence. AB - BACKGROUND AND OBJECTIVES: Medication mismanagement is a serious health issue affecting elders and people with disabilities, who often manage multiple medications. This project's goal was to educate medical and nursing students about the challenges patients face when managing complex medication regimens. METHODS: A total of 104 first-year medical students and 40 second-year nursing students were randomly assigned to participate in a 1-week regimen of mock prescriptions or to read a description of the regimen and make predictions about what the experience would be like had they participated. RESULTS: Quantitative results in combination with qualitative information suggest that the students taking the mock prescriptions gained important insights into the difficulty of managing a complicated medication regimen. DISCUSSION: This mock prescription exercise, well accepted by students and faculty, was easily incorporated into the curriculum and provided an experiential opportunity for students to learn of the difficulties of medication adherence. PMID- 19184687 TI - Medical students, money, and career selection: students' perception of financial factors and remuneration in family medicine. AB - BACKGROUND AND OBJECTIVES: Medical students have had a declining interest in family medicine as a career. Some studies have shown a small inverse relationship between debt levels and primary care, but it is unclear how students perceive remuneration in different specialties and how these perceptions might influence career choice. METHODS: Medical students at one school were surveyed to understand their perceptions of physician remuneration and to gain insight into how these perceptions might affect career selection. RESULTS: Response rate was 72% (560/781 students). Students' estimates of physician income were accurate throughout training, with the overall estimate for family medicine being lower than the actual income by only $10,656. The vast majority of students agreed with the statement that family physicians get paid too little (85%-89% of each class). The importance of payment as a factor in career decision making increased with higher debt and with advancing training. CONCLUSIONS: Students are able to accurately predict income by specialty from an early stage of training and have a negative perception of income in family medicine. The perception that family physicians make too little money could be an important driver--or at least a modifier--in the lack of interest in family medicine. PMID- 19184688 TI - A step ahead--evaluating the clinical judgment skills of incoming interns. AB - BACKGROUND: Assessing the baseline competency of incoming interns is a challenge to faculty of all residency programs. This article describes and evaluates the Intern Clinical Judgment Evaluation, a tool developed to test the clinical judgment and communication skills of incoming interns. METHODS: During orientation week, each intern is evaluated using structured clinical scenarios modeled after "on call" situations. Interns are given feedback on information collection, medical decision making, judgment, understanding of the medical system, and written and verbal communication skills. Data from 4 years is presented, comparing predictions of performance from faculty and from the Intern Clinical Judgment Evaluation to actual year-end intern performance data. RESULTS: There was excellent agreement between the Intern Clinical Judgment Evaluation and the summary of interns' performance (kappa value=0.8). Faculty predictions were a less accurate predictor of intern performance (kappa value=0.6). The Intern Clinical Judgment Evaluation had a sensitivity of 93% (13/14), specificity of 88% (15/17), positive likelihood ratio (LR+) of 7.9, and negative likelihood ratio (LR-) of 0.08. Comparatively, faculty predictions of resident performance had a sensitivity of 57% (8/14), specificity of 94% (17/18), LR+ of 10.3, and LR- of 0.40. Surveyed interns felt the evaluation process was a positive experience. CONCLUSIONS: The Intern Clinical Judgment Evaluation was a valid tool to predict which interns would require more faculty supervision during intern year. Based on solicited feedback from tested interns, it also was a positive testing experience. PMID- 19184689 TI - Improving the health of diabetic patients through resident-initiated group visits. AB - BACKGROUND AND OBJECTIVES: Group visits have the potential to benefit patients with chronic illnesses. Our objective was to implement resident-run diabetic group visits that would improve patient education and help patients become more involved in their care. METHODS: We developed systems to promote, coordinate, and lead the visits. Residents' responsibilities were delegated through a preparation checklist. A standardized progress note was developed to encourage patient goal setting and to track relevant laboratory test results. To evaluate our program, we conducted surveys to determine patients' behavioral changes and satisfaction levels and assessed the effect on group visit participants' glycated hemoglobin (HbA1c) and low-density lipoprotein (LDL) levels. RESULTS: Group visit patient survey results showed that 72% of the patients in the group visit cohort reported making a lifestyle change; 88% felt that the group visit helped them achieve better control of their diabetes, and 100% stated that they would come to a group visit again. However, no significant changes were noted in HbA1c or LDL levels. CONCLUSIONS: Residents can overcome challenges and implement, organize, and run effective group visits that increase patients' self-reported self-management abilities, but we could demonstrate no statistically significant improvement on measurable biochemical parameters of glucose or lipid control. PMID- 19184690 TI - Factors related to publication success among faculty development fellowship graduates. AB - BACKGROUND AND OBJECTIVES: This study's purpose was to understand factors related to scholarly project publication among fellowship graduates and to explore the role of collegial relationships and institutional characteristics on ever having published and number of publications. METHODS: We surveyed 5 years of graduates (2000-2004) from the Michigan State University Faculty Development Fellowship Program (n=90) via e-mail. The survey instrument included questions about barriers and facilitators, colleague relationships, and aspects of their institutional environment, the latter adapted from a validated survey by Bland et al. RESULTS: The response rate was 70%. Only seven graduates had published the results of their fellowship project; 44% had published at least one paper. Barriers to publishing scholarly project results included lack of time and assistance, inability to finish projects, and rejection of a submitted paper. Factors associated with publication included types of collegial relationships and a number of institutional factors in addition to the Bland score. CONCLUSIONS: Few of our fellowship graduates published the results of their scholarly project, and less than half published papers. We identified multiple barriers and facilitators that could be addressed by the fellowship programs and home institutions to enhance publication success. PMID- 19184692 TI - Essays and commentaries in family medicine. The human side of health care. PMID- 19184691 TI - Patient age influences perceptions about health care communication. AB - OBJECTIVE: The study's objective was to determine if a patient's age is independently associated with how he/she perceives interactions with health care providers. METHODS: We used a secondary, cross-sectional analysis of nationally representative data from the 2002 Medical Expenditure Panel Survey (MEPS). We measured the independent association between patient age and six outcomes pertaining to communication and decision-making autonomy, while simultaneously controlling for gender, race, ethnicity, family income, educational attainment, census region, rural residence, insurance status, and usual source of care. RESULTS: Compared to patients>or=65 years, patients ages 18-64 were less likely to report that their provider "always" listened to them, "always" showed respect for what they had to say, and "always" spent enough time with them. DISCUSSION: Patient perceptions of health care interactions vary by age. A better understanding of how and why age is associated with patient-provider communication could be useful to design practice-level interventions that enhance services and also to develop national policies that improve health care delivery and health outcomes. PMID- 19184693 TI - A process for setup and development of an online portfolio with user input. PMID- 19184694 TI - How IMCI can be integrated into medical school curriculum and impact health. PMID- 19184695 TI - The delphi technique as a needs assessment for family medicine curriculum. PMID- 19184696 TI - Intracellular levels of hydrogen peroxide and nitric oxide in oocytes at various stages of meiotic cell cycle and apoptosis. AB - The objective was to find out the functional roles of hydrogen peroxide (H(2)O(2)) and nitric oxide (NO) during various stages of meiotic cell cycle and apoptosis in rat oocytes. For this purpose, 30 oocytes from each stage such as diplotene, metaphase-I (M-I), metaphase-II (M-II) and apoptosis were collected and intracellular H(2)O(2), total nitrite level and inducible nitric oxide synthase (iNOS) expression were analysed. This study demonstrated that generation of a tonic level of H(2)O(2) induces meiotic resumption in diplotene-arrested oocytes and further increase may lead to apoptosis. Conversely, reduction in iNOS expression and total nitrite level are associated with meiotic resumption in diplotene-arrested oocytes, but induce apoptosis in aged oocytes. These results suggest that generation of a tonic level of H(2)O(2), reduced iNOS expression and total nitrite level are associated with meiotic resumption, while more generation of H(2)O(2) and sustained reduced total nitrite level are linked with oocyte apoptosis in rat. PMID- 19184697 TI - Computational approaches may underestimate pK(a) values of longer-chain perfluorinated carboxylic acids: Implications for assessing environmental and biological effects. AB - Acidity constants were calculated using the semiempirical PM6 pK(a) estimation method for all C(2) through C(9) perfluoroalkyl carboxylate (PFCA) congeners and the straight-chain C(10) through C(13) isomers. According to the PM6 estimates, the linear congeners within each PFCA homologue group have the highest pK(a) values by up to 6 units depending on the degree of branching in the perfluoroalkyl chain. In general, the higher the degree of branching in the perfluoroalkyl chain within a homologue group, the lower the estimated pK(a) value. When the branching is closest to the terminal carboxylate group, the effect on the calculated pK(a) is greatest. Although the PM6 calculated pK(a) values agree well with previously reported estimates for selected linear PFCA congeners using the SPARC and COSMOtherm approaches, all computational approaches only show good agreement with reported experimental values for short chain PFCAs (C(2) through C(5)). Increasing divergences are observed between calculated and experimental results by up to several pK(a) units as the perfluoroalkyl chain length increases beyond C(5). The findings demonstrate a need for additional experimental pK(a) measurements for an expanded set of both linear and branched PFCA congeners to confirm previous experimental reports that are potentially in error, and upon which to calibrate existing computational methods and environmental, toxicological, and waste treatment method models. PMID- 19184698 TI - Activated sludge versus aerated lagoon treatment of kraft mill effluents containing beta-sitosterol and stigmasterol. AB - In the present study the performance of an activated sludge system (AS) and an aerated lagoon (AL) are compared for the treatment of kraft cellulose effluents. Both reactors were operated at organic loading rates (OLR) up to 9 g COD/L. d. The activated sludge system presented better performance at high OLR than the aerated lagoon with removal percentages of 57-67% and 95% of COD and BOD(5), respectively. The removal percentages of specific compounds were always slightly higher in the case of the AS than in the AL with exception of the total phenolic compounds which concentration increased in the former. Furthermore the analyzed fractions from the effluents indicated that in the AS the compounds were fully removed while in the case of the aerated lagoon were transformed into compound with low molecular weight P2 (5 kD < MW < 10 kD). In the case of the AS system the increase of the total phenolic compounds concentrations was corroborated with the increase of the fractions P2, P3 and P4. The studied phytosterol were removed from the effluent at concentrations up to 2 mg/L in percentages close to 100% mainly by adsorption in the case of the AL and by a combining adsorption and mainly degradation in the AS. The BOD(5) concentration in the effluent of both systems fulfilled the requirements of the Chilean regulations for effluent discharge indicating the feasibility of aerobic treatment of this kind of effluents. PMID- 19184699 TI - Determination of Ct values for chlorine of resistant enteroviruses. AB - Information on the required chlorine dose or Ct value (concentration of free chlorine multiplied by contact time) is limited to only a few enteroviruses. In this study the Ct values of some of the reported more chlorine resistant enteroviruses were determined. The Efficiency Hom Model was used to predict the times for 2, 3. and 4 -log inactivation of echovirus 1 and 12, coxsackievirus B5 and poliovirus type 1 at pH 7.5 and 9 at 5 degrees C. Coxsackievirus B5 was the most resistant to chlorine with a Ct requirement of 11.5 mg x min/L at pH 7.5 at 5 degrees C compared to polio with a Ct of 5.3 mg x min/L under the same conditions. All the viruses were more resistant than polio 1 at pH 9.0. PMID- 19184700 TI - Permeation sampling of phthalate esters in water. AB - A method for sampling the six phthalate esters considered priority pollutants by the United States EPA has been developed. The sampling device utilizes a silicone polycarbonate membrane (SSP-M213) through which the analytes permeate and are collected onto an adsorbent, Amberlite XAD-16. After a known period, the samplers were removed from the water and the analytes were extracted from the adsorbent using dichloromethane/hexane (50:50, v/v), with the resulting solution analyzed using gas chromatography with flame ionization detection. Time-weighted average (TWA) concentrations of the phthalates in the water were determined by creating plots of amount of analyte collected versus the product of concentration and time (ppm. hr) for each phthalate. A linear correlation between the amount of analyte collected and the product of concentration and time was seen for each phthalate. Effects of temperature, stirring rate and the presence of potential interferents on the rate of permeation were also studied. The obtained and minimal amounts of solvent are used for sample preparation and injection into the gas chromatograph. PMID- 19184701 TI - Biogrouting compared to jet grouting: environmental (LCA) and economical assessment. AB - In order to predict consequences of replacing jet grouting with biogrouting, and identify major contributors to the cost of both technologies, a large road project in Stockholm, Sweden, was used as a case study. Jet grouting had been used to seal the contact between sheet piling and bedrock, biogrouting for the same function was computed. A comparative environmental and economical assessment was carried out using life cycle assessment (LCA). The results show that biogrouting was cheaper than jet grouting and would have had lower environmental impact. The major difference was the transport and use of heavier equipment for jet grouting. Biogrouting also used less water and produced less landfilled waste. However, the production of urea and CaCl(2) for biogrouting required much energy. PMID- 19184702 TI - Removal of arsenate and 17alpha-ethinyl estradiol (EE2) by iron (hydr)oxide modified activated carbon fibers. AB - Activated carbon fibers (ACF) were modified with iron (hydr)oxide and studied to determine their suitability to remove arsenate and 17alpha -ethinyl estradiol (EE2) from water. Two synthesis methods, one involving aqueous KMnO(4) pretreatment followed by Fe(II) treatment, and the other involving reaction with Fe(III) in an organic solvent followed by NaOH treatment, were used to produce modified ACF media containing 5.9% and 8.4% iron by dry weight, respectively. Scanning electron microscopy (SEM) and Electron dispersion X-ray (EDX) techniques indicated slightly higher iron content near the outer edges of the fibers. Pseudo equilibrium batch test experimental data at pH = 7.0 +/- 0.1 in 5 mM NaHCO(3) buffered ultrapure water containing approximately 100 micro g(As)/L and approximately 500 micro gEE2/L were fitted with the Freundlich isotherm model (q = K x C(E)(1/n)). The adsorption capacity parameters (K) were approximately 2586 (micro gAs/gFe)(L/micro gAs)(1/n) and approximately 425 (micro gAs/gFe)(L/micro gAs)(1/n)), respectively, for the KMnO(4)/Fe(II) and Fe(III)/NaOH treated media. The KMnO(4)/Fe(II) media exhibited a lower adsorption capacity at 99% EE2 removal than did the Fe(III)/NaOH treated media (1.3 mgEE2/g -dry -media vs. 1.8 mgEE2/g dry -media). The arsenate adsorption intensity parameters (1/n) for both modified ACF media were < 0.29, implying very favorable adsorption, which suggests that this type of media may be suitable for single point -of -use applications in which arsenic and organic co-contaminants require simultaneous removal and the depth of the packed bed is the key factor. PMID- 19184703 TI - Effect of phosphate and sediment bacteria on trichloroethylene dechlorination with zero valent iron. AB - The effects of sediment-isolated bacteria and phosphate on the efficacy of zero valent iron (ZVI) for the dechlorination of trichloroethylene (TCE) were examined in batch experiments. TCE (0.3 mM) and a constant concentration of sediment bacteria were simultaneously exposed to ZVI in the presence of 0 mmol, 15 mmol, and 30 mmol of phosphate. TCE profiles, starting from 0.3 mM to about 0.1 mM, exhibited two-phase of sorption kinetics at all three phosphate concentrations without the sediment bacteria. TCE removal was less and slower with phosphate in the system. With the sediment bacteria, however, more TCE was removed with the sediment bacteria than without it, unlike our initial hypothesis. With the sediment bacteria and phosphate, the concentration of ferrous (0.505 mM) ions was doubled that with phosphate only (0.271 mM). The sediment bacteria in this research, mainly Bacillus sp., could contribute to the long-term stability of ZVI reactivity for dechlorination of TCE in sediment. The sediment bacteria in this research could reduce the iron or chelate the evolved ferrous ions to retain the reducing reactivity of ZVI. PMID- 19184704 TI - Concentration of trace elements in human semen and relation to spermatozoa quality. AB - In this study the concentrations of trace elements such as lead, cadmium, iron, nickel, copper and zinc in the human semen (n = 47), occurrence of pathological spermatozoa, and correlations of these elements to pathological forms were investigated. For each sample of human spermatozoa at least 500 spermatozoa were evaluated. Metal contents were determined by the voltametric method and flame absorption spectrophotometry method. The concentrations of trace elements in human semen were: lead 1.49 +/- 0.40 mg x kg(- 1), cadmium 0.13 +/- 0.15 mg x kg( 1), iron 2.59 +/- 0.21 mg x kg(- 1), nickel 0.40 +/- 0.07 mg x kg(- 1), copper 0.28 +/- 0.06 mg x kg(- 1), and zinc 153.93 +/- 67.08 mg x kg(- 1), respectively. The total percentage of pathological spermatozoa was 41.61 +/- 9.80% with predominancy of broken flagellum, flagellum torso and separated flagellum. In relation to trace elements the analysis showed correlation between copper and lead (r = -0.47), nickel and iron (r = 0.36), lead and flagellum ball (r = 0.39), cadmium and large heads (r = 0.37) and between iron and other forms of pathological spermatozoa (r = -0.32). Results of this study describe possible effects of trace elements on the spermatozoa quality in normal human sperm. PMID- 19184705 TI - Mercury contamination and potential impacts from municipal waste incinerator on Samui Island, Thailand. AB - In recent years, mercury (Hg) pollution generated by municipal waste incinerators (MWIs) has become the subject of serious public concern. On Samui Island, Thailand, a large-scale municipal waste incinerator has been in operation for over 7 years with a capacity of 140 tons/day for meeting the growing demand for municipal waste disposal. This research assessed Hg contamination in environmental matrices adjacent to the waste incinerating plant. Total Hg concentrations were determined in municipal solid waste, soil and sediment within a distance of 100 m to 5 km from the incinerator operation in both wet and dry seasons. Hg analyses conducted in municipal solid waste showed low levels of Hg ranging between 0.15-0.56 mg/kg. The low level was due to the type of waste incinerator. Waste such as electrical appliances, motors and spare parts, rubber tires and hospital wastes are not allowed to feed into the plant. As a result, low Hg levels were also found in fly and bottom ashes (0.1-0.4 mg/kg and Mg > Zn > Cu > Ni and physicochemical indicators decreased: conductivity > HMF > moisture > pH > a(w). PMID- 19184711 TI - Progress and challenges in numerically modelling solid sports balls with application to softballs. AB - Much of the work surrounding finite element simulation of bat-ball impacts has focused on techniques describing the ball. Determining the accuracy of these models has been hindered by challenges in experimentally characterizing the ball's response. In the following, dynamic mechanical analysis and an instrumented impact test were used to characterize the solid ball at deformation rates representative of play. The ball was described in the numerical model as a linear viscoelastic material. It was observed that a Prony series model based on small deformation dynamic mechanical analysis did not provide sufficient energy loss upon impact, while a simpler Power Law model, fitted to large deformation data, described the measured energy loss and impact force over a range of speeds. Results of a parametric study are presented as a guide towards tailoring the parameters of the Power Law model to match the measured energy loss and impact force. Discrepancies observed between the experiment and the numerical model suggest that the ball response should be characterized in environments closely resembling game conditions. PMID- 19184712 TI - Choose your primary outcome variables with care. PMID- 19184713 TI - A time-motion analysis of professional rugby league match-play. AB - The aim of this study was to analyse movement patterns of professional rugby league players during matches played as part of Australia's National Rugby League (NRL) competition. The movement patterns of one player from each of the three positional groups (hit-up forwards, adjustables, and outside backs) during three competition matches were examined using time-motion analysis. Positional groups differed in distances covered (P = 0.001), and covered shorter distances than those described in earlier research. The mean exercise-to-rest ratio was 1:6 for the outside backs and hit-up forwards and 1:5 for the adjustables. However, such ratios did not reflect the most demanding periods of the game, which included repeated high-intensity efforts interspersed with recovery periods of short duration. These periods of repeated high-intensity exercise often occurred at crucial phases of the game, when players were either attacking or defending the try-line. Furthermore, patterns of movement during repeated high-intensity periods of play differed among positional groups. To prepare for the most highly intense periods of match-play, professional rugby league players should adopt position-specific training that includes the highest and lowest exercise-to-rest ratios likely to be experienced in competition. PMID- 19184714 TI - A novel allele of HWP1, isolated from a clinical strain of Candida albicans with defective hyphal growth and biofilm formation, has deletions of Gln/Pro and Ser/Thr repeats involved in cellular adhesion. AB - Gene HWP1 encodes a major Candida albicans hyphae cell wall protein which is a substrate of mammalian transglutaminases, promoting the cross-link of the fungus to epithelial cells. Here, we describe a novel HWP1 allele, isolated from C. albicans blood isolates. Analysis of the translated sequence shows that three important regions are absent in the novel allele, HWP1-2, relative to the previously described allele, HWP1-1. Regions 1 and 2 consist of 10 amino acid repeats important for functional conformation of peptide chains and attachment of C. albicans cells to the mammalian epithelia. Region 3 consists of 34 amino acid residues rich in threonine and serine, with O-glycosylation sites that promote the cross-linking with other proteins on C. albicans surface. The HWP1-2 homozygous strain L757 and the heterozygous strain L296 (HWP1-1/HWP1-2) have significantly lower levels of HWP1 expression during hyphal growth and biofilm formation compared to strain SC5314 (HWP1-1/HWP1-1). However, strain L296 properly forms hyphae and biofilms in vitro while strain L757 has reduced hyphal growth (40.4%) and biofilm formation (90.8%). Our results indicate that the HWP1 locus has biofilm specific allelic differential expression and suggest that the HWP1-2 encoded protein is less efficient to maintain cell-to-cell and cell-to surface adhesion during biofilm formation. This is the first report of a natural variant of HWP1. PMID- 19184718 TI - Mortality of the oldest old Chinese: the role of early-life nutritional status, socio-economic conditions, and sibling sex-composition. AB - Based on a nationally representative sample of 8,099 Chinese drawn from the Chinese Longitudinal Healthy Longevity Survey (CLHLS), this study investigated the long-term health consequences of early-life nutritional status, sibling sex composition, childhood socio-economic conditions, and place of birth on mortality at ages 80 and above between 1998 and 2005. Better nutritional status in childhood predicted lower mortality at ages 80 and above, net of childhood circumstances, adult socio-economic status, and health behaviours. In addition, sibling sex-composition had long-term health consequences, net of childhood and adult characteristics, such that women benefited from having grown up in families with only daughters, while men benefited from having grown up in families with both sons and daughters. Childhood socio-economic status was only marginally related to old-age mortality and this association was attenuated further by the inclusion of adult characteristics. Place of birth was not a significant predictor of old-age mortality. PMID- 19184719 TI - Effect of early conditions on disability among the elderly in Latin America and the Caribbean. AB - Poor early conditions have been associated with increasing risks of some chronic diseases during adulthood. Since chronic illnesses are known to be important risk factors for disability, poor early conditions should predict disability at older ages. In addition, recent literature suggests that poor early conditions may affect the risk of disability even in the absence of chronic illnesses. We aimed to evaluate the magnitude of differentials in the risk of being disabled according to early conditions experienced by elderly populations in Latin America and the Caribbean, and to identify the group of chronic illnesses responsible for it. We find that poor early conditions exert a strong influence on disability later in life in two ways: by increasing the risk of suffering disability-related chronic illnesses and by increasing the risks of suffering disabilities by those with chronic illnesses. PMID- 19184720 TI - Effect of HIV/AIDS-related mortality on household dependency ratios in rural South Africa, 2000-2005. AB - With data from a surveillance system that uses verbal autopsies to identify cause of death in rural South Africa, we investigated whether mortality from HIV/AIDS differs from other causes of death in its effect on household dependency ratio, and to what extent the effect is mediated by the baseline dependency ratio. FINDINGS: (i) the impact of death from HIV/AIDS on the dependency ratio in 2005 is marginally positive compared with other causes of death, but (ii) the impact is overpowered by the effect of death at working age, and (iii) the baseline dependency ratio mediates the effects on the 2005 ratio of cause of death and of the individual's sex and age at death. Migration into and out of the household- anticipating or responding to a death--seems to be a key source of change in the household dependency ratio. PMID- 19184721 TI - Family division in China's transitional economy. AB - Using a longitudinal data-set (the China Health and Nutrition Survey) we explored the effect of various economic factors, including household wealth, employment sector, and involvement in a household business on the division of extended families in China's transitional economy. Results from event history analyses suggest that these economic factors act as either a dividing or a unifying force on the extended family. Household wealth reduces the risk of family division, but the effect is weaker for families in which parents have upper secondary education. In addition, an extended family is more likely to divide when married children work in the state sector. Further, the probability of family division is higher in families where daughters-in-law work in the state sector than in those with sons in this sector. Finally, involvement in a household business for married children increases family stability. PMID- 19184722 TI - Influences of the family of origin on the timing and quantum of fertility in the Netherlands. AB - This study examined to what extent family of origin influences age at first birth and final number of children. We took into account direct intergenerational transmission of fertility behaviour, family-life experiences around age 15, and the wider social context of the family of origin. Hazard regression analyses (n=6,630) and Poisson regression analyses (n=3,736) were performed using data from the 2002-03 wave of the Netherlands Kinship Panel Study. Besides a positive relation between parents' and their children's fertility patterns, we find that positive experiences of family life around age 15 encourage childbearing: the less conflict between parents experienced by the child during early adolescence, the younger the adult child at first birth, and the larger the number of the adult child's children. The number of children is also influenced positively by frequency of contact with non-residential relatives during early adolescence. The socio-economic status and religiosity of the parental family also affect fertility. PMID- 19184723 TI - The determinants of discrimination against daughters in China: evidence from a provincial-level analysis. AB - This paper reports a provincial-level analysis of the way in which various socio economic and socio-demographic determinants influence the decision to discriminate against daughters in China. While most existing studies use the infant or child sex ratio as the only variable to be explained, this study analyses separately the two main discriminatory practices: sex-selective abortion (with sex ratio at birth as a proxy) and neglect of girls' health care (with excess infant mortality among females as a proxy). The analysis helps to illuminate the circumstances that encourage sex-selective behaviours, which appear to be dictated mainly by extreme poverty, family support to the elderly, and father's education, together with the social pressure on couples to adhere to traditional values and roles and the constraints on family size. While sex selective abortion appears to result from long-term strategies to optimize family composition, lethal neglect is the immediate result of economic constraints. PMID- 19184725 TI - Volatile organic compounds and formaldehyde as explaining factors for sensory irritation in office environments. AB - This study's database comprised results of volatile organic compound (VOC) measurements from 176 office buildings. In 23 of the 176 buildings, formaldehyde measurements were also conducted. It was suspected that the buildings had indoor air problems, but a walk-through inspection did not reveal any clear, abnormal contaminant sources. The 50 most abundant VOCs and their concentrations in 520 air samples were analyzed. The irritation potency was estimated for 33 out of the 50 common VOCs and their mixtures, as well as for formaldehyde. This information was used to calculate the recommended indoor air levels (RILs) for the VOCs. The RILs were considerably higher than the measured mean indoor air concentrations in the buildings. However, the RIL for formaldehyde was exceeded in most of the 23 buildings studied. According to the evaluation of irritation potency, formaldehyde was a more likely cause of sensory irritation than the mixture of common nonreactive VOCs at the concentrations that occurred in the buildings without abnormal indoor sources. Furthermore, environmental symptoms of office workers were characterized in 20 office buildings (including the database of 176 office buildings) with the aid of an indoor air questionnaire. The most frequent symptoms related to the indoor environment were involved the upper respiratory tract. However, no relationship could be shown between the reported symptoms and the occurrence of VOC and formaldehyde concentrations in these buildings. Generally, the study results indicated that formaldehyde was the more likely agent causing sensory irritation than the mixture of the common nonreactive VOCs at the concentrations occurring in the buildings without abnormal indoor sources. PMID- 19184726 TI - 2nd Norwegian Environmental Toxicology Symposium: joining forces for an integrated search for environmental solutions. PMID- 19184727 TI - Transcriptional regulation in liver and testis associated with developmental and reproductive effects in male zebrafish exposed to natural mixtures of persistent organic pollutants (POP). AB - Persistent organic pollutants (POP) occur as mixtures in nature and it is difficult to predict the toxicity of such mixtures based on knowledge about toxicity and mechanisms of action for single compounds. The present knowledge on the combined toxic effects and modes of actions of exposure to mixtures is limited. Thus, the scientifically based hazard and risk assessment of POP requires analytical and toxicological data from studies with environmental mixtures of POP. The application of genome wide transcription profiling in toxicology, in combination with classical endpoints, will improve the current understanding of the mechanisms of toxic processes. Furthermore, gene expression data may be useful in establishing new hypothesis and discovering new biomarkers for known toxicity as well as not yet recognized toxicity endpoints. In the present study, developmental and reproductive effects of lifelong exposure to environmental relevant concentrations of two natural mixtures of POP were investigated using classical and molecular methods in a controlled zebrafish model. The mixtures used were extracted from burbot (Lota lota) liver originating from freshwater systems in Norway: one mixture with high levels and one mixture with background levels of polybrominated diphenyl ethers (PBD), polychlorinated biphenyls (PCB), and DDT. The concentration of POP in the zebrafish ranged from levels detected in wild fish from Lake Mjosa, to concentrations reported in human and wildlife populations. Phenotypic effects observed in both exposure groups included (1) reduced survival, (2) earlier onset of puberty, (3) increased male/female sex ratio, and (4) differences in body weight at 5 mo of age. Interestingly, genome-wide transcription profiling showed changes in regulation of genes involved in endocrine signaling and growth. The transcriptomics changes included (1) key regulator genes for steroid and thyroid hormone functions (cga, ncoa3), (2) insulin signaling and metabolic homeostasis (pik3r1, pfkfb3, ptb1), and (3) p53 activation (mdm4). The effects observed in the experimental zebrafish model raise the question of whether chemical pollution represents a risk to the reproductive health of wild fish inhabiting the freshwater system. PMID- 19184728 TI - Gene expression of GST and CYP330A1 in lipid-rich and lipid-poor female Calanus finmarchicus (Copepoda: Crustacea) exposed to dispersed oil. AB - The copepod Calanus finmarchicus is a marine ecological key species in the Northern Atlantic food web. This species was exposed to an artificially weathered North Sea oil dispersion (oil droplets and water-soluble fractions [WSF]) and a filtered dispersion (containing only WSF) in serial dilution. Female copepods were divided into lipid-rich and lipid-poor for each exposure followed by gene expression analyses of glutathione S-transferase (GST) and cytochrome P-450 330A1 (CYP330A1). Lipid-rich copepods exhibited elevated transcription of GST and reduced transcription of CYP330A1 after exposure to both dispersed oil and WSF. In contrast, lipid-poor copepods exhibited increased transcription of CYP330A1 following exposure to WSF but not the dispersion. Data suggested that small lipid storage promotes increased bioavailability of accumulated oil compounds. Variations in response in CYP330A1 gene expression indicate that oil constituents may exert different modes of toxic action in copepods depending on their reproductive stages. The contribution of oil droplets to the observed effects seemed to be low as GST gene expression was similar after exposure to both dispersed oil and WSF. However, feeding rate in copepods exposed to dispersed oil was reduced, and this may have decreased the uptake of oil constituents via the diet. Although quantitatively higher mortality was observed in copepods exposed to the highest dispersion levels, this may result from smothering of animals by oil droplets. Furthermore, increasing dilution of both the dispersions and the WSF altered their distributions and chemical composition, which may influence the bioavailability of spilled crude oil to pelagic marine organisms. PMID- 19184729 TI - Are Atlantic cod in Store Lungegardsvann, a seawater recipient in Bergen, affected by environmental contaminants? A qRT-PCR survey. AB - The aim of this study was to examine the transcriptional levels of selected genes in liver and head kidney of Atlantic cod Gadus morhua sampled in Store Lungegardsvann, a seawater recipient situated in the middle of the city of Bergen, Norway, for effects of contaminants released from municipal sewage effluents and former dump sites. Five males and six females were caught with fish traps in Store Lungegardsvann in 2006. Cod from a location near Jondal in the Hardanger Fjord were used as controls (five males and four females). The following 12 genes were picked as potential markers of contaminant exposure: cytochrome P-450 1A (CYP1A), cytochrome P-450 2C33-like (CYP2C33-like), cytochrome P-450 3C (CYP3C), glutathione S-transcriptase pi (GST) (detoxification and biotransformation), Mn superoxide dismutase (Mn SOD), glutathione reductase (GR), heat-shock protein 70 (HSP70) (oxidative stress), vitellogenin A (VtgA), vitellogenin B (VtgB), zona pellucida 2 (ZP2) (effects of estrogen disruptors), B cell lymphoma 2 (Bcl-2), and cyclin-dependent kinase inhibitor 1A (CDKN1A) (radiation). The results showed that two males caught in Store Lungegardsvann possessed high transcriptional levels of VtgA, VtgB, and ZP2 mRNA in the liver. In addition, CYP1A was 4.9-fold higher expressed in males from Store Lungegardsvann compared to males from the reference population. CYP2C33-like mRNA expression was significantly higher (1.8-fold) in females from Store Lungegardsvann than in females from the reference population. CYP1A was significantly lower (4.7-fold) expressed in head kidney of females from Store Lungegardsvann than in females from Hardanger Fjord. In a follow-up examination with sexually mature cod sampled in Store Lungegardsvann in 2007, the livers were shown to contain high levels of polychlorinated biphenyls (PCB) and dioxin-like PCB. In conclusion, fish inhabiting Store Lungegardsvann are exposed not only to endocrine disruptors but also to other contaminants that affect the transcription of phase I biotransformation genes. PMID- 19184730 TI - Induction of DNA double-strand breaks in the H4IIE cell line exposed to environmentally relevant concentrations of copper, cadmium, and zinc, singly and in combinations. AB - Xenobiotics, including heavy metals, exist in nature as complex mixtures of compounds with possible interactions. Induction of DNA damage such as DNA strand breaks may exert detrimental consequences to both individuals and populations. In this study, the induction of DNA double-strand breaks was assessed using the H4IIE rat hepatoma cell line following exposure to high and environmentally relevant concentrations of chloride salts of the metals cadmium (Cd), copper (Cu), and zinc (Zn), both singly and in combination. DNA strand break analysis was performed using agarose gel electrophoresis. Median molecular lengths were calculated from fragment size distributions acquired from gel image data and were used as a quantitative measure of DNA double-strand break induction. Exposure to high concentrations of Cu and Cd in combination produced a significant increase in the occurrence of DNA strand break. However, exposing cells to high concentrations of Cu, Cd, and Zn in combination resulted in significantly lower DNA double-strand break compared to control cells. Addition of low Zn to the Cd/Cu mixture restored DNA damage level back to that of the control. Environmentally relevant concentrations of Cd, Cu, and Zn did not appear to induce DNA strand breaks in the H4IIE cell line. PMID- 19184731 TI - Altered stress-induced cortisol levels in goats exposed to polychlorinated biphenyls (PCB 126 and PCB 153) during fetal and postnatal development. AB - Short-term stress exposure is associated with activation of the hypothalamic pituitary-adrenal (HPA) axis and a consequent rise in blood glucocorticoids and catecholamines, from the adrenal cortex and medulla, respectively. The HPA axis is a potential target for some persistent organic pollutants, among which polychlorinated biphenyls (PCB) were found to be modulators of the mammalian endocrine system. PCB are distributed globally in the environment, in food chains, and are transferred to the fetuses of pregnant animals and via mother's milk to suckling offspring. In the present study it was postulated that intrauterine and lactational exposure to either of two single congeners of PCB (PCB 153 and PCB 126, respectively) might affect basal cortisol concentrations, and also the cortisol response to short-term stress in adulthood. Thus, pregnant goats were orally exposed to one of these PCB congeners from d 60 of gestation until delivery, and their offspring studied. Low-dose exposure to PCB 153 and PCB 126 resulted in significantly lower mean basal cortisol concentrations in goat offspring during certain periods of pubertal development and their first breeding season. Male goat kids exposed to either PCB congener showed a greater and more prolonged rise in plasma cortisol levels than controls when animals were subjected to mild stress at 9 mo of age using frequent blood sampling. Neither the basal maternal cortisol plasma level nor goat kid adrenal masses were affected by PCB exposure. PMID- 19184732 TI - Vitellogenin in African sharptooth catfish (Clarias gariepinus): purification, characterization, and ELISA development. AB - Vitellogenin (Vtg) induction in African sharptooth catfish (Clarias gariepinus) was assessed in order to develop a method for monitoring estrogenic pollution in African freshwater systems. Clarias gariepinus Vtg (Cg-Vtg) was purified from serum obtained from 17alpha-ethynylestradiol (EE2)-exposed fish and polyclonal antibodies against Cg-Vtg were raised. An enzyme-linked immunosorbent assay (ELISA) was developed and the induction and kinetics of Vtg were assessed in male fish in three different exposure trials using both natural estrogen (17alpha estradiol [E2]) and synthetic EE2. Concentrations of EE2 in water and levels of EE2 conjugates in bile were quantified by liquid chromatography-mass spectrometry (LC-MS). In addition, co-administration of E2 and benzo[a]pyrene (BaP) were studied. Vtg was induced in all exposure trials and the maximum induction was observed 1 wk after exposure. Exposure of male C. gariepinus to 1.4, 2.7, and 13.9 microg/ml EE2 induced Vtg synthesis at all concentrations. BaP did not influence the Vtg kinetics. However, an increased rate of biliary excretion of EE2 was observed when BaP was additionally administered. In conclusion, Vtg is induced in male C. gariepinus after exposure to both E2 and EE2, rendering it a suitable biomarker for endocrine-disrupting chemicals in African freshwater systems. PMID- 19184733 TI - Androgenic modulation of early growth of Atlantic cod (Gadus morhua L.) previtellogenic oocytes and zona radiata-related genes. AB - Available evidence suggests that androgens play critical roles in early oocyte growth and development in fish. However, the molecular mechanisms underlying this important aspect of reproductive endocrinology have not yet been established. In this study the effects of androgens (11-ketotestosterone [11-KT] and testosterone [T]) were determined on gene expression patterns and growth of cod previtellogenic oocytes, using an in vitro oocyte culture technique. Previtellogenic ovarian tissue was cultured for 5 and 10 d at different concentrations of 11-KT and T (0, 1, or 1000 microM) dissolved in ethanol (0.3%). The androgen concentrations were selected as they represent physiological and supra-physiological concentrations, respectively. Quantitative polymerase chain reaction (PCR) demonstrated increased mRNA expression for five genes recently identified as androgen responsive in our subtracted cDNA library in previtellogenic cod ovary exposed in vitro to androgens. Quantitative histological analyses showed a consistent stereological validation of oocyte growth and development after exposure to androgens. In general, both 11-KT and T induced previtellogenic oocyte growth and development, and these effects were more pronounced with 11-KT exposure. Taken together, our study reveals some novel roles of androgens on the development of previtellogenic oocytes, indicating control of early follicular and oocyte growth in cod ovary. The potent effects of 11-KT on oocyte growth support our earlier hypothesis that non-aromatizable androgens play significant roles in regulating early oocyte growth with potential consequences for the fecundity process. Therefore, these novel roles of androgens as promoters of ovarian growth and development presented in this study may be useful for the aquaculture industry and for breeding of new captive and endangered species. From a toxicological point of view, the cod is a marine species and exposure to complex chemical mixtures that may exert androgenic and/or anti-androgenic effects represents an environmental issue of reasonable concern in the marine environment. Therefore, the findings in the present study represent a novel basis that can be used to determine the effects of xenoandrogens on oocyte development and fecundity in this important marine species. PMID- 19184734 TI - Biomarker responses in mussels, an integrated approach to biological effects measurements. AB - Biological effects techniques have been used with the aim to further integrate biological effects measurements with chemical analysis and apply these methods to provide an assessment of mussel health status. Live native mussels were collected from selected coastal and estuarine sites around the British Isles, including the rivers Test, Thames, Tees, and Clyde, and Lunderston Bay. A suite of biological effects techniques was undertaken on these mussels, including whole organism responses (scope for growth), tissue responses (histopathology), and subcellular responses (lysosomal stability, multi-xenobiotic resistance [MXR], and Comet assay). In addition, whole mussel homogenates were used to measure organic (polycyclic aromatic hydrocarbons [PAH], polychlorinated biphenyls [PCB]) and metal concentrations. Overall the mussels collected from the Thames were in relatively poor health, based on histopathological markers, significantly higher DNA damage, and elevated expression of MXR detoxifying proteins. In contrast, the mussels collected from the River Test were in the best health, based on histopathological markers, respiration rate (SFG), and low frequency of DNA damage. In conclusion, the biological effects techniques were able to distinguish between relatively contaminated and clean environments, with the Thames mussels in worst health. Mussel tissue chemistry data were not able to explain the variations in biological response. Evidence indicates that the difference in the health of the mussels between the different sites was due to either effects of contaminants that were not measured, or the combined effects of mixture toxicity resulting in a threshold effect. PMID- 19184735 TI - Effects of tributyltin (TBT) on in vitro hormonal and biotransformation responses in Atlantic salmon (Salmo salar). AB - The mechanisms by which the biocide tributyltin (TBT) and its metabolites affect the hormonal and xenobiotic biotransformation pathways in aquatic species are not well understood. In this study hepatocytes isolated from salmon were used to evaluate the mechanistical effects of TBT on fish hormonal and xenobiotic biotransformation pathways. Cells were exposed to 0.01, 0.1, 1, or 5 microM TBT and samples were collected at 0, 12, 24, or 48 h following exposure. Gene expression patterns were evaluated using quantitative polymerase chain reaction (PCR), and cytochrome P-450 (CYP)-mediated enzyme activities were evaluated by ethoxyresorufin, benzyloxyresorufin, and pentoxyresorufin O-deethylase (EROD, BROD, and PROD, respectively) activity assays. Generally, exposure of hepatocytes to 1 microM (at 48 h) and 5 microM TBT (at 12, 24, and 48 h) consistently produced reductions in all mRNA species investigated. TBT produced significant decreases of vitellogen (Vtg) expression at 48 h and modified the expression patterns of estrogen receptors (ERalpha and ERbeta) and androgen receptor-beta (ARbeta) that were dependent on time and TBT concentration. In the xenobiotic biotransformation pathway, TBT produced differential expression patterns that were dependent on exposure time and concentration for all salmonid AhR2 isoforms (AhR2alpha, AhR2beta, AhR2delta, and AhR2gamma). For CYP1A1, CYP3A, AhRR, and Arnt mRNA, TBT produced exposure- and time-specific modulations. Catalytic CYP activities showed that BROD activity increased in an apparent concentration specific manner in cells exposed to TBT for 12 h. Interestingly, EROD activity showed a TBT concentration-dependent increase at 24 h and PROD at 12 and 48 h of exposure. In general our data show that TBT differentially modulated hormonal and biotransformation responses in the salmon in vitro system. The apparent and consistent decrease of the studied responses with time in 1 and 5 microM exposed hepatocytes suggest a possible transcription inhibitory effect of TBT. PMID- 19184736 TI - Xenopus tropicalis as a test system for developmental and reproductive toxicity. AB - The usefulness of Xenopus tropicalis as a model species to investigate endocrine disruption and developmental reproductive toxicity was assessed. In our test system tadpoles were exposed to test substances from shortly after hatching until metamorphosis, including the period of gonadal differentiation. Effects on the sex hormone and thyroid hormone axes were evidenced as skewed sex ratios, malformations of reproductive organs, altered cytochrome (CYP19) (aromatase) activity, and gene expression in gonads and brain, as well as changed thyroid histology and time to metamorphosis. Reproductive toxicity was evaluated at sexual maturity. Male-to-female sex reversal was implied at concentrations as low as 6 pM (1.8 ng/L) ethynylestradiol (EE2), which is comparable to EE2 levels observed in the environment. EE2-exposed males that were not sex reversed had significantly reduced fertility and a reduced amount of spermatozoa in testes compared with control males. This indicates that reproduction in wild frogs might be impaired by estrogenic environmental pollutants. Aromatase activity in brain and testes of adult frogs was not affected by larval EE2 exposure. Preliminary results indicate that exposure to the environmentally relevant pharmaceutical clotrimazole modulated aromatase activity in brain and gonads during sex differentiation, which warrants further investigation. The susceptibility to estrogen-induced sex reversal of X. tropicalis was comparable to that of other frog species and fish. Similarities between the reproductive effects in X. tropicalis and those reported in fish, birds, and mammals after developmental exposure to estrogens make X. tropicalis promising model for research on endocrine disruption and developmental reproductive toxicity. PMID- 19184737 TI - Relationships between physiology, tissue contaminants, and biomarker responses in Atlantic cod (Gadus morhua L.). AB - It is generally assumed that there should be some relationship between tissue residues of contaminants and related biomarker responses in the same individual, with a possible additional modulation through physiological processes. The main basis for such assumptions originated from experimental studies, but it has been less easy to detect and quantify any such relationships in field-collected material. The aim of the current study was to clarify whether the responses of selected biomarkers (cytochrome P-4501A [CYP1A], delta-aminolevulinic acid dehydratase [ALA-D], metallothionein [MT]) could be correlated with physiological parameters and/or contaminant levels. Atlantic cod (Gadus morhua L.) were sampled over a decade at a number of locations along the Norwegian coast. The locations included both polluted and reference areas. In addition to contaminant-related responses (biomarkers), physiological characteristics were registered for each individual and the contaminant load determined. There were clear relationships between responses in all three biomarkers and tissue contaminants, but additional factors such as age or size and liver lipid or liver somatic index also contributed in multiple-regression models. The results indicated that responses in all three biomarkers reflected relevant, contaminant-related processes in the cod even though none of them provided a simple separation between polluted and pristine locations. PMID- 19184738 TI - Relationship between polycyclic aromatic hydrocarbon (PAH) accumulation in semipermeable membrane devices and PAH bile metabolite levels in Atlantic cod (Gadus morhua). AB - The extent to which accumulations of polycyclic aromatic hydrocarbons (PAH) by semipermeable membrane devices (SPMD) may be correlated to the concentration of PAH metabolites in fish was examined. Atlantic cod (Gadus morhua) and SPMD were exposed over a 4-wk period to a mixture that was designed to simulate the composition of PAH in produced water discharges in the North Sea. Fish bile and whole SPMD were analyzed after exposure in a high, low, and a control treatment tank. PAH metabolites were detected and quantified by means of high-performance liquid chromatography (HPLC) or gas chromatography-time-of-flight mass spectroscopy (GC-ToF-MS) and PAH in SPMD by gas chromatography-mass spectroscopy (GC-MS). The level and duration of exposure were generally reflected in both matrices. Exposure water concentrations, calculated from SPMD accumulations, showed some considerable differences from nominal concentrations for volatile compounds. Concentration factors (CF) for fish based on SPMD-derived water concentrations ranged from 2.9 to 354.3 L/g. CF increased with hydrophobicity with more variation at higher log K(ow). Regression analysis of accumulations of individual compounds in fish and SPMDs showed reasonable but compound-specific correlation, r2 = .54-.85 (C1-phenanthrenes/anthracenes and pyrene, respectively). Comparison of the two exposures suggests that fish CF may not be entirely independent of water concentrations. Overall the results show that accumulation of up to four-ring PAH in SPMD may be related to internal exposure of fish to these substances when exposed through water, as evidenced by metabolite concentrations. The usefulness of using SPMD to measure exposure concentrations in a large-scale flow-through study is also examined. PMID- 19184739 TI - Cellular energy allocation in Hediste diversicolor exposed to sediment contaminants. AB - Environmental contaminants released into water tend to accumulate in sediment, leading to exposure for sediment-dwelling organisms. This study aimed to clarify whether chronic (56 d) exposure to a mixture of environmentally relevant concentrations of contaminants (tributyltin [TBT] and perfluorononanoic acid [PFNA]) spiked in clean sediment would affect the sediment-dwelling polychaete Hediste diversicolor. Endpoints were burial time, biomass change, and cellular energy allocation (CEA). Sediments were spiked with these two contaminants to produce different levels of contamination. Chemical analyses of the sediment following the exposure indicated that the spiking procedure was only partially successful and treatments were grouped into four categories according to contaminant concentrations: "background," "low," "moderate," and "high." No significant differences were found between treatments with respect to burial time, biomass change, or energy storage fractions (proteins, lipids, carbohydrates) in the polychaetes. A significant increase in cellular respiration was found for polychaetes in "moderate" and "high" treatment groups compared to the "background" treatment group. This resulted in significantly lower cellular energy allocation values for "moderate" and "high" treatment groups compared to "background." Although the effects were not marked, the results show that sediment-dwelling organisms may be affected following chronic exposure to environmental contaminants. There is a need to clarify whether chronic exposures exert a negative effect on sediment-dwelling organisms. PMID- 19184740 TI - Polycyclic aromatic hydrocarbon (PAH) metabolites in Atlantic cod exposed via water or diet to a synthetic produced water. AB - Bile metabolites are widely accepted as measures of exposure to polycyclic aromatic hydrocarbons (PAH) and have also been used to assess exposure to alkyl phenols (AP). The aim of this study was to clarify relationships between exposure (through water or diet) and subsequent accumulation of specific PAH and AP metabolites in Atlantic cod (Gadus morhua). Atlantic cod were exposed through water or diet to a synthetic mixture of PAH, alkylated PAH, and AP, simulating the composition of North Sea produced water (formation water; separated from oil or gas on the platform). Fish were exposed through water for 11 mo and the results reported here are from 2 and 8 mo. Fish were subjected to one of four exposures: "control," "low," "high," or "pulsed high." Bile samples were analyzed by gas chromatography/time-of-flight mass spectroscopy (GC/MS-ToF) for identification of PAH and AP metabolites. Cod exposed through diet were divided into six groups receiving different groups of compounds (AP, PAH, or alkylated PAH), a mixture of all compounds, a low-concentration mixture, and controls. A dose-dependent relationship was found for metabolites for most of the PAH and AP, although results were less clear for the more volatile substances such as phenol and naphthalene. The concentration of bile metabolites from fish exposed through water rose with increasing lipophilicity, but this relationship was less clear for fish exposed through diet. Overall, data indicated that bile metabolites of AP and PAH in fish are reliable markers of both water exposure and dietary exposure to such substances, although with the possible exception of the more volatile species such as phenol and naphthalene. PMID- 19184741 TI - Immunotoxicity of polychlorinated biphenyls (PCB) in free-ranging gray seal pups with special emphasis on dioxin-like congeners. AB - Increased pinniped and dolphin mortality rates have led to speculations that persistent pollutants, in particular polychlorinated biphenyls (PCB), are immunomodulatory, making individuals susceptible to infections. The aim of the present study was to investigate effects that PCB may exert on peripheral blood mitogen-induced lymphocyte proliferation responses in free-ranging gray seal (Halichoerus grypus) pups from the polluted Baltic Sea and from the cleaner open waters of the Atlantic Ocean. T-cell mitogen (phytohemagglutinin [PHA] and concanavalin A [Con A]) and T-/B-cell mitogen (pokeweed mitogen [PWM]) responses were significantly lower in the more PCB-contaminated Baltic gray seal pups than in reference pups from the Atlantic. Best regression analysis revealed that dioxin-like mono-ortho PCB accounted for the immunosuppressive responses. In the Baltic seals, the immunosuppressive effects were limited to the more persistent dioxin-like mono-ortho PCB. In the Atlantic seals a positive correlation existed between PCB concentrations and mitogen-induced lymphocyte proliferation responses. Although direct cause-effect relationships are difficult to demonstrate in free-ranging animals, our results are consistent with the weight of evidence available that PCB, and dioxin-like PCB in particular, modulate lymphocyte function in free-ranging marine mammals. Thus, exposure to PCB most likely result in increased susceptibility to infections or reduced host resistance in pinnipeds. PMID- 19184742 TI - Effect of water hardness on the toxicity of cadmium to the green alga Pseudokirchneriella subcapitata in an artificial growth medium and nutrient spiked natural lake waters. AB - The effect of cadmium (Cd) on the growth rate of the green alga Pseudokirchneriella subcapitata was investigated at three different levels of water hardness in an artificial growth medium and in three nutrient spiked, Norwegian natural soft lake waters. The lake water with the lowest levels of hardness and dissolved organic carbon (DOC) (Lake Byglandsfjorden) showed the highest toxicity attributed to Cd. In water from this lake without addition of calcium and artificial chelators, the EC50 for inhibition of growth rate was 9.4 microg Cd/L and EC10 was 2.8 microg Cd/L. When the hardness of the waters was increased by addition of calcium the adverse effects of Cd were in general reduced. In all tested media, the concentration/effect slope was reduced with increasing hardness. This led to a higher effect of hardness on EC50 than EC10. PMID- 19184743 TI - Local sources of polychlorinated biphenyls (PCB) in Russian and Norwegian settlements on Spitsbergen Island, Norway. AB - Samples of surface soil, flaking paint, concrete, transformer oils, and small capacitors were collected from the three largest coal-mining settlements on Spitsbergen--Barentsburg (Russian), Pyramiden (Russian), and Longyearbyen (Norwegian)--to study the role of potential local sources of polychlorinated biphenyls (PCB) in the arctic areas (78 degrees N). Median concentrations of PCB7 in soil from Barentsburg and Pyramiden were 0.268 and 0.172 mg/kg, respectively, with a maximum concentration of 28.7 mg/kg. High concentrations found in paint (3520 mg/kg) and small capacitors (114,000 mg/kg) indicated that these two are the main sources of local PCB contamination. Only traces of PCB were found in the Longyearbyen samples compared to the results from the other two settlements. Large amounts of building refuse, electrical waste, and scrap metals constitute major pollution sources in Barentsburg and Pyramiden. Weathering and general decay facilitate the mobilization of PCB from these sources to the local soil, which consequently is readily available for fluvial and eolian transport to the more vulnerable marine environment. PMID- 19184744 TI - Effect of every-other-day fasting on spontaneous chromosomal damage in rat's bone marrow cells. AB - Dietary restriction in experimental rodents, either by calorie restriction (CR) or by every-other-day fasting (EODF), was shown to protect against cancer and increase lifespan. One of the suggested hypotheses to explain the beneficial effects of dietary restriction is that the diet stabilizes the integrity of the genetic information. The effects of EODF on the spontaneous frequency of sister chromatid exchanges (SCE) and chromosomal aberrations (CA) were examined in bone marrow cells of 3-mo-old Wistar male rats. After 12 wk of EODF diet, significant reduction in the frequency of SCE and total number of CA was observed. Data indicate a protective effect of EODF diet against spontaneous mutations in rats. PMID- 19184745 TI - Environmental mutagens induced transversions but not transitions in regulatory region of mitochondrial DNA. AB - One of the long-term objectives of the research in our laboratory was to determine whether mitochondrial DNA (mtDNA) mutations were generated in cell lines exposed to a variety of known mutagens. Many of these mutagens are known to increase oxidative stress in the cell, and one potential outcome of this would be an increased incidence of point mutations in mtDNA. Recently, there has been some controversy regarding the validity of point mutations in the regulatory region of mtDNA as a predictive or causative marker for carcinogenesis. Studies were undertaken to assess whether nuclear mutagens such as arsenic (As), asbestos, and ultraviolet (UV) and gamma-radiation, induced both heteroplasmic and homoplasmic point mutations in mtDNA. A direct sequencing approach was used to reduce the occurrence of experimental errors and cross-checked all base changes with databases of known polymorphisms. Our results showed that, while base changes did occur, there was no marked difference between the number of changes in treated and untreated cells. Furthermore, in human lymphocyte samples from subjects exposed to As, most of these base changes were previously reported. Interestingly, there was an increase in the number of transversions (purine ( pyrimidine) in smokers from a human population study, but as with the findings in cell culture samples, there was no difference in the total number of base changes. Data suggest that only a change in the number of rare transversions would be indicative of an increase in point mutations in mtDNA after exposure to mutagens. PMID- 19184746 TI - The effect of weathering on ecopersistence, reactivity, and potential toxicity of naturally occurring asbestos and asbestiform minerals. AB - The mechanisms underlying asbestos toxicity mainly rely on experiments performed on "laboratory" fibers, but little data is available on naturally occurring asbestos (NOA). Human exposure to NOA is subject to their ecopersistence and the modulation of their potential toxicity following weathering. The effect of weathering on three fibrous minerals from the Italian Western Alps, chrysotile, tremolite, and balangeroite-a Fe-rich asbestiform mineral-was investigated by mimicking more than 100 yr of physical (freezing-thawing/wetting-drying cycles in a climatic chamber) and biochemical forces (incubation with oxalic acid). Ion release, evaluated by means of inductively coupled plasma-atomic emission spectroscopy (ICP-AES), and variation in chemical composition, evaluated by scanning electron microscopy coupled with energy-dispersive spectroscopy (SEM EDS), indicated that weathering modified the fibers in the series: chrysotile > balangeroite > tremolite. Kinetics of ion release from the fibers (Mg, Fe, and Si) revealed different ion removal pathways. Tremolite was poorly affected. Chrysotile preferentially released cations up to a plateau, with physical and biochemical forces acting competitively. Conversely, for balangeroite, upon which weathering forces acted synergistically, the initial loss of ions facilitated further dissolution and more Si than Mg was released, suggesting an ongoing collapse of the crystal structure. Depletion of redox-reactive ions produced a significant reduction in fiber-derived *OH radicals (EPR, spin-trapping technique), but the fibrous nature was always retained. Despite weathered fibers appearing less toxic than "stored/laboratory" ones, NOA is to be considered far from safe because of fibrous nature and residual surface reactivity. Risk assessment needs to consider the effect of weathering on exposures. Both tremolite and balangeroite may contaminate, in some areas, chrysotile asbestos. However, in contrast to tremolite, balangeroite exhibits a low ecopersistence, similar to chrysotile behavior. Any contribution of balangeroite to chrysotile toxicity will thus be related to its quantitative occurrence and not to higher structural stability. PMID- 19184747 TI - The usual suspects-influence of physicochemical properties on lag time, skin deposition, and percutaneous penetration of nine model compounds. AB - The influence of physicochemical properties of nine model compounds on lag time, skin deposition, and percutaneous penetration was evaluated. Static diffusion cells mounted with human skin were used as the experimental model, and experiments were carried out in accordance with Organization for Economic Cooperation and Development (OECD) guidelines. The model compounds were chosen to cover a wide spectrum of solubilities and molecular weights. The pesticides included were glyphosate, dimethoate, pirimicarb, malathion, paclobutrazol, methiocarb, prochloraz, and benzoic acid, with the ninth model compound being caffeine. The fastest dermal penetration was observed for compounds with log Pow values between 1.5 and 4. Malathion did not fit into this generalization. No clear relationship was observed between molecular weight and Kp values. The shortest lag time was observed for the most hydrophilic model compounds. With increasing molecular weight, the lag time rose. Thus, the lag time for the smallest model compound was close to 1.5 h, while the lag time exceeded 20 h for a model compound with a molecular weight of 377 g. A difference in lag time of this magnitude inevitably produces differences in the amounts of a chemical able to penetrate the skin within a limited period of time. The relative deposition in the skin was highest for the lipophilic model compounds. For log Pow values between -1 and 2, a linear relationship was observed between log Pow and log Kp. Comparisons between theoretical Kp values based on the Potts-Guy equation and experimental Kp values demonstrated good agreement. Experimental Kp values based on experiments using full-thickness human skin generally underestimated the calculated Kp values by 25%. The observations are relevant to industrial hygienists and regulatory agencies using experimental data for assessing systemic toxicity following dermal exposures as for development of structure activity relationships for dermal absorption of chemicals. PMID- 19184748 TI - Effects of Asian dust storm events on hospital admissions for congestive heart failure in Taipei, Taiwan. AB - In spring, windblown dust storms originating in the deserts of Mongolia and China make their way to Taipei city. These occurrences are known as Asian dust storm (ADS) events. These ADS events lead to enhanced PM(10) levels over those generated by the usual local sources. The objective of this study was to assess the possible associations of PM(10) with hospital admissions for congestive heart failure (CHF) in Taipei, Taiwan, during the period from 1996 to 2001. Fifty-four dust storm episodes, which were classified as index days, were identified. Daily CHF admissions on index days were compared with admissions on comparison days. Two comparison days were selected for each index day, 7 d before the index days and 7 d after the index days. The effects of ADS on hospital admissions for CHF were prominent 1 d after the event (relative risk = 1.114; 95% confidence interval = 0.993-1.250). However, the association was not statistically significant. There may not have been enough power to detect associations resulting from the inadequate sample size of CHF admissions on ADS events days. However, it seems worthwhile to pay more attention to the ADS events and health in the future. PMID- 19184749 TI - Cancer mortality in chinese populations surrounding an alloy plant with chromium smelting operations. AB - This report is a further characterization of data from an ecological cancer mortality study of a population (about 10,000) exposed to groundwater contaminated by hexavalent chromium [Cr(VI)] up to 20 mg/L near JinZhou City in the LiaoNing Province of China between 1960 and 1978. Prior reports showed an elevation in all-cancer mortality from 1970 to 1978 averaged across five agricultural villages with Cr(VI) in groundwater relative to average cancer rates for the district and province. The current study compares the cancer rates during the same time period for the same five exposed villages to those of four nearby areas with no Cr(VI) in groundwater. The use of a local comparison group is considered superior to the use of district or province averages because of the expected improved similarity among unmeasured covariates in nearby areas. The average lung-, stomach-, and all-cancer mortality rates for the three agricultural villages without Cr(VI) in groundwater were not statistically different from those of the five agricultural villages with Cr(VI) in groundwater. Also, three surrogate measures of village drinking-water Cr(VI) dose did not significantly correlate with cancer mortality rates in the five exposed villages. Further, the industrial town in which the Cr(VI) source was located had different demographics and a different pattern of stomach and lung cancers compared to the adjacent agricultural villages, regardless of Cr(VI) groundwater exposure. The results of other local investigations on cancer mortality and genotoxicity in the exposed populations are reviewed. The overall findings in the studied population do not indicate a dose-response relationship or a coherent pattern of association of lung-, stomach-, or all-cancer mortality with exposure to Cr(VI)-contaminated groundwater. PMID- 19184750 TI - Cytotoxicity screening for the cyanobacterial toxin cylindrospermopsin. AB - The cell lines C3A, HepG2, NCI-87, HCT-8, HuTu-80, Caco-2, and Vero were screened for sensitivity to the cyanobacterial toxin cylindrospermopsin (CYN), with the aim of determining the most sensitive cells to be used in cytotoxicity tests. Cell lines were chosen to be representative of the organs targeted by the toxin; liver, kidney, intestine, and were expected to have different metabolic activities and uptake capabilities. Over the range of cell lines tested, IC(50) determinations at 24 h (MTT assay) ranged fourfold, from 1.5 muM for hepatocyte derived cell lines (C3A IC(50) = 1.5 +/- 0.54; HepG2 IC(50) = 1.5 +/- 0.87) to 6.5 +/- 3.3 micro the colon-derived Caco-2 cell line. The cell-line sensitivity seemed to decrease in cell lines derived from progressively more distal regions of the gastrointestinal tract: gastric > duodenal > ileal > colonic. The greater sensitivity of the hepatic cell lines to CYN was also apparent in 7-d exposure studies, with low toxin concentrations exerting cytotoxic effects that were not seen in other cell lines. Short-term exposure of C3A cells to CYN (1-6 h) was shown to induce cytotoxicity at 24 h despite a washout and recovery incubation, demonstrating the protracted and apparently irreversible nature of CYN's toxic effects. PMID- 19184751 TI - Percutaneous absorption of 3,3',4,4'-tetrachlorobiphenyl (PCB 77) from soil. AB - Six dermal absorption experiments (one in vivo, five in vitro) were conducted using 3,3',4,4'-tetrachlorobiphenyl (TCB) either neat at 141 microg/cm2 or sorbed on a low organic (LOS) or high organic (HOS) soil at 6-10 microg/cm2. All soil experiments were conducted at 1000 ppm and soil loads of 6-10 mg soil/cm(2). After 96 h the percentage of applied dose absorbed (PADA) for TCB sorbed on LOS was 49.7 (rat, in vivo), 31.9 (rat, in vitro), and 7.4 (human, in vitro). The 96 h PADA for TCB sorbed on HOS was 9.6% (rat, in vitro). Generally, rat skin was observed to be four- to ninefold more permeable to TCB than human skin (in vitro). At steady state, the dermal flux of TCB on LOS at 1000 ppm and on HOS at 1000 ppm (both in vitro, rat) was 33 and 10 ng/cm2/h, respectively (ratio = 3.3). PMID- 19184753 TI - Crises of commitment: ethics of intimacy, kin, and confinement in global psychiatry. AB - This article addresses the challenges inherent to framing an ethical language around the issue of patient "abandonment" in psychiatric hospitals. It describes the situations of divorced women in a north Indian private clinic to ask how a Foucaultian understanding of "the family" as a site of discipline, and the ethical languages that emerge from this critique, cannot account for the complexities of kin life in these women's lives and the multiple legal domains that shape them as subjects. PMID- 19184754 TI - Precipitating pharmakologies and capital entrapments: narcolepsy and the strange cases of Provigil and Xyrem. AB - Since the mid-1990s, Americans have been made more aware of chronic sleep deprivation and sleep disorders exacerbated by dominant temporal regimes of work, school, and family life, primarily through increased medical and media attention. Concomitantly, Americans have turned to medical treatments and pharmaceutical cocktails to achieve normalcy rather than attending to the social and cultural causes of sleep sickness. This turn toward pharmaceuticalization is aided in part by the proliferation of medical disorders and the pharmaceuticals marketed to treat them (e.g., "excessive daytime sleepiness" requires treatment once reserved for narcoleptics). These cocktails have explicit and implicit components: the former consist of pharmaceuticals, the latter of capital dependencies, including ties to medical insurance companies, stable employment, and familial networks. In this article, I examine the proliferation of pharmaceutical cocktails through the concept of the pharmakon-something simultaneously remedy and cause-to illuminate the causes and effects of such pharmaceutical regimens in contemporary American society, specifically those relating to sleepiness. Specific cases of this struggle between chemical dependence and normalcy are offered from my ethnographic work with patients who suffer from sleep disorders. PMID- 19184755 TI - Transcultural medicine: a multi-sited ethnography on the scientific-industrial networking of Korean medicine. AB - Through a multi-sited ethnography of three different types of organization--a traditional medical clinic, two laboratories, and a biotech company--this article examines how Korean medicine (KM) scientizes, globalizes, and industrializes its clinical knowledge. By tracing the complex networking process among multiple places, I aim to understand how KM reinvents its knowledge, identity, and boundaries in a global situation. In particular, I pay attention to how this process involves multiple dimensions of power relations, economic interests, and scientific authorities. This article concludes that heterogeneous and unequal encounters between KM, science, and industry lead to simultaneous productions of new culture and power without reducing them to a single logic or center in a global age. PMID- 19184756 TI - Labor, social exclusion, and chronic muscular illness: the case of mid impoverished sectors in a peripheral neighborhood in Madrid, Spain. AB - This study analyzes the combination of different and differing representations and care practices that social groups belonging to a mid-impoverished sector of Madrid use to alleviate chronic muscular pain, as well as the consequences that this has for both domestic and work life. I collected empirical evidence in a peripheral neighborhood of Madrid during 2004 using a number of anthropological methods such as participant observation, in-depth interviews, and life-history interviews. The following results can be singled out from the completed investigation: in the context of social impoverishment there are macrostructural factors that are transformed into health-destructive processes that influence the development and incidence of chronic muscular illness. This requires the different social groups that are suffering from this to establish a number of care practices mainly based on the use of Western medicine as well as other medical traditions of self-care. These practices relate to economic, educational, sociocultural, and ideological factors that characterize the lives of these individuals. This essay focuses on perceptions of illness and care practices beyond the biomedical context, as it addresses those representations and practices that these impoverished social groups display in accordance with their way of life and in relation to their ailments. PMID- 19184757 TI - Foot soldiers of global health: teaching and preaching AIDS science and modern medicine on the frontline. AB - This article investigates the ways in which global health messages and forms of health citizenship are mediated by AIDS activists in rural South Africa. It focuses on how these activists and treatment literacy practitioners are not only concerned with changing the lives of people living with AIDS to better manage biological conditions associated with their seropositive status, but also with how they are also committed to recruiting new members into their biopolitical projects and epistemic communities. These mobilization processes involve translating and mediating biomedical ideas and practices into vernacular forms that can be easily understood and acted on by the "targets" of these recruitment strategies. However, these processes of "vernacularization" of biomedical knowledge often occur in settings where even the most basic scientific understandings and framings of medicine cannot be taken for granted. This ethnographic case study shows that global health programs and their local mediators often encounter "friction" from the most powerful national actors as well as the most marginalized local ones. PMID- 19184758 TI - Caries, gender and socio-economic change in the Xavante Indians from Central Brazil. AB - BACKGROUND: The oral health conditions of indigenous peoples in Amazonia are closely associated with ecological and dietary changes related to interaction with non-Indians. AIM: The study investigated the incidence of caries in an indigenous community from Central Brazil focusing on gender differences. SUBJECTS AND METHODS: The research was conducted among the Xavante Indians and was based on longitudinal data collected in two surveys (1999 and 2004). The study included 128 individuals, 63 (49.2%) males and 65 (50.8%) females, divided in four age brackets (6-12, 13-19, 20-34, 35-60 years of age). The DMFT (decayed, missing and filled teeth) index and incidences (difference between 1999 and 2004) were calculated for each individual. The proportion of incidence was also calculated. Differences in caries risk between gender and age brackets were compared by parametric and non-parametric tests. RESULTS: There were statistically significant differences in relation to caries incidence between age brackets and gender. The greatest incidence was observed in the 20-34 age bracket, which presented 3.30 new decayed teeth, twice the risk of the 6-12 age bracket (p<0.01), chosen as reference. While females in most age groups did not show higher risk for caries when compared to males, there was a 4.04-fold risk in the 20-34 age bracket (p<0.01). CONCLUSION: It is concluded that factors related to the social functions of each sex (gender issues) and differential access to information, health services, and education may help to understand the differences observed in the incidence of caries. PMID- 19184759 TI - Protective role of 27bp repeat polymorphism in intron 4 of eNOS gene in lacunar infarction. AB - Association of the three potential endothelial nitric oxide synthase gene (eNOS) polymorphisms (T-786C in promoter region, G894T in exon 7 and tandem 27-bp repeats in intron 4) with an increased risk of lacunar infarction (LI) were investigated. Genotypes of 70 patients and 81 healthy controls were determined through PCR with or without RFLP. Flow-mediated dilatation (FMD) was performed to assess endothelial-dependent vasodilatation, whereas the endothelial-independent vasodilatation was assessed with nitroglycerin (NTG). Genotype distribution was significantly different between LI patients and controls for intron 4aa (alleles for four repeats), genotype frequency being 1.4% and 16.0%, respectively (odds ratio for additive effect, 0.47; 95% CI, 0.28-0.81; p=0.006). Haplotypes with the intron 4aa polymorphism were significantly higher in controls when compared with the LI group (p=0.001). Diminished FMD but normal NTG response confirmed that patients with LI have generalized endothelial dysfunction. Intron 4aa genotype of eNOS gene seems to be protective for isolated LI and the effect was potentiated by the absence of 786C polymorphism in any allele of the promoter region. PMID- 19184760 TI - Elevated oxidative stress, iron accumulation around microvessels and increased 4 hydroxynonenal immunostaining in zone 1 of the liver acinus in hypercholesterolemic rabbits. AB - Rabbits were fed a diet containing 1% cholesterol for 8 weeks and the levels of iron and oxidized lipids in liver analysed using atomic absorption spectroscopy and gas chromatography-mass spectrometry. A non-significant trend to an increase in iron level, but significant increases in the lipid peroxidation products, F(2) isoprostanes and the cholesterol oxidation products 7 beta hydroxycholesterol, 7 ketocholesterol and cholesterol 5,6-alpha epoxide were detected in the liver of the cholesterol-fed rabbits. Histological analysis showed greater accumulation of lipids by Sudan red labelling in hepatocytes of zone I than zones II and III of the liver acinus. The increase in lipids coincided with an increase in iron staining in macrophages around liver microvessels and increased immunostaining to melanotransferrin and the lipid peroxidation product, 4-hydroxynonenal (4-HNE), in zone 1. The results are suggestive of microvascular damage associated with iron accumulation and oxidative stress in the liver during hypercholesterolemia. PMID- 19184761 TI - Effects of cereal bars for breakfast and mid-morning snacks on mood and memory. AB - The aim of the present study was to examine the effects of consuming cereal bars, given either for breakfast or a mid-morning snack, on mood and memory. Thirty-two volunteers (16 males, 16 females; mean age, 20 years 9 months) were randomly assigned to one of four groups formed by combining breakfast (cereal bar versus no breakfast) and snack (cereal bar versus no snack) conditions. A baseline session was completed at 08:30 h followed by breakfast at 9:00 h, another test at 10:00 h, followed by a mid-morning snack and then a final test at 12:00 h. In each session, volunteers rated their mood and carried out four memory tasks: free recall; recognition memory; a verbal reasoning task; and a semantic processing task. The results showed that volunteers who consumed a cereal bar for breakfast felt more alert, happy and sociable and less anxious. In addition, they also recalled more words in a free recall task. When the cereal bar was consumed as a mid-morning snack, alertness and hedonic tone increased, especially in the group who received no breakfast. The group who had no breakfast reported reduced anxiety after consumption of the snack. Recall was also improved after the snack. These findings show that consuming cereal bars in the early and mid-morning leads to beneficial behavioural effects. The results confirm earlier research on effects of breakfast and extend our knowledge of effects of snacks. Consumption of cereal bars may have important practical applications especially in situations where preparation of breakfast is difficult. PMID- 19184762 TI - Effects of cartilage polysaccharide on apoptosis of human hepatoma BEL-7402 cells and murine H22 hepatocarcinoma. AB - OBJECTIVE: To detect the mechanism of apoptosis on hepatoma BEL-7402 cells induced by cartilage polysaccharide in vitro and the effect on H22 hepatocarcinoma mice in vivo. MATERIALS AND METHODS: TUNEL assay, flow cytometry, immunofluorescent staining and animal experiments were used to examine the effects of cartilage polysaccharide on human hepatoma BEL-7402 cells in vitro or H22 hepatocarcinoma mice in vivo. RESULTS: We found that cartilage polysaccharide had a dramatic effect on inducing apoptosis in BEL-7402 cells in a dose-dependent and time-dependent manner. The molecular mechanism of the process of apoptosis was investigated, which was correlated with FAS-caspase and regulated the expression level of Vimentin and p21 protein. The survival rate and lifespan of the experimental group were better than for the model group. CONCLUSION: Our data demonstrate that cartilage polysaccharide may serve as a novel source of anti hepatocarcinoma agent that may play an important role in future clinical trials. PMID- 19184764 TI - Validation of the folate food frequency questionnaire in vegetarians. AB - Adequate folate status has an important role in the prevention of chronic and developmental diseases and is considered a potential public health issue. Therefore, valid tools for measuring the vitamin intake are needed. In our previous study a folate food frequency questionnaire (FFQ) designed to measure dietary folate equivalents was developed and validated among adult women against serum and erythrocyte (red blood cell) folate and plasma homocysteine. The aim of the present study was to validate the FFQ in vegetarians (n=75). The Pearson correlation for folate intake and biomarkers was 0.41, 0.36 and -0.15 for serum and red blood cell folate and plasma homocysteine, respectively. The quadratic weighted kappa value for biomarkers was above 0.2 and the gross misclassification of subjects into quartiles was less than 10%. The FFQ is a valid tool for measuring dietary folate equivalent intake in Croatian vegetarians. PMID- 19184765 TI - Validation of the folate food frequency questionnaire with serum and erythrocyte folate and plasma homocysteine. AB - The aim of the present study was to develop and validate a folate food frequency questionnaire (FFQ)1 designed to measure dietary folate equivalents. The self administered FFQ containing 39 items and a reference period of 1 month (i.e. the previous month), was validated against three biomarkers: serum and erythrocyte (RBC) folate, and plasma homocysteine (Hcy). Subjects were women (n=99) between the ages of 21 and 87 years. The Pearson correlation coefficients for folate intake and biomarkers were 0.36, 0.34 and -0.25 for serum and RBC folate, and plasma Hcy, respectively. A principal component factor analysis was performed on the three biomarkers to calculate the folate status factor. The Pearson correlation for the folate status factor and folate intake was 0.39. The FFQ described in this study is a valid tool for measuring folate intake expressed as dietary folate equivalents in adult women and is suitable for future investigations about the relationship between folate and disease, or as an educational tool. PMID- 19184766 TI - A testable prognostic model of nicotine dependence. AB - Individuals' dependence on nicotine, primarily through cigarette smoking, is a major source of morbidity and mortality worldwide. Many smokers attempt but fail to quit smoking, motivating researchers to identify the origins of this dependence. Because of the known heritability of nicotine-dependence phenotypes, considerable interest has been focused on discovering the genetic factors underpinning the trait. This goal, however, is not easily attained: no single factor is likely to explain any great proportion of dependence because nicotine dependence is thought to be a complex trait (i.e., the result of many interacting factors). Genomewide association studies are powerful tools in the search for the genomic bases of complex traits, and in this context, novel candidate genes have been identified through single nucleotide polymorphism (SNP) association analyses. Beyond association, however, genetic data can be used to generate predictive models of nicotine dependence. As expected in the context of a complex trait, individual SNPs fail to accurately predict nicotine dependence, demanding the use of multivariate models. Standard approaches, such as logistic regression, are unable to consider large numbers of SNPs given existing sample sizes. However, using Bayesian networks, one can overcome these limitations to generate a multivariate predictive model, which has markedly enhanced predictive accuracy on fitted values relative to that of individual SNPs. This approach, combined with the data being generated by genomewide association studies, promises to shed new light on the common, complex trait nicotine dependence. PMID- 19184768 TI - An increasing trend of cutaneous zygomycosis caused by Mycocladus corymbifer (formerly Absidia corymbifera): report of two cases and review of primary cutaneous Mycocladus infections. AB - Cutaneous zygomycosis is a rare but serious infection in trauma patients. We report two cases of cutaneous zygomycosis caused by Mycocladus corymbifer (formerly Absidia corymbifera) which were probably the result of soil contamination of wounds of the patient's lower extremities. Both patients received appropriate antifungal therapy in combination with aggressive surgical debridement. While a cure was achieved with amphotericin B in one, the other patient was intolerant to this antifungal and cure was achieved with a new drug, posaconazole. Twenty seven cases (including the two cases in this study) of cutaneous M. corymbifer zygomycosis reported in the literature were reviewed. The data showed an increase in infections with 16 cases (59.2%) reported since 2002. PMID- 19184769 TI - Systemic phaeohyphomycosis due to Exophiala (Wangiella) in an immunocompetent child. AB - We report a rare case of systemic lymphadenitis and hepatic involvement due to Exophiala (Wangiella) dermatitidis in a pediatric patient. An 8-year-old immunocompetent boy with chronic fever was examined through the use of sonography and CT scan which demonstrated cervical and mesenteric lymph node enlargement and numerous small hepatic lesions. The etiologic agent was isolated by means of lymph node aspiration. The fungus was identified by its morphological characteristics and through DNA sequencing of the internal transcribed spacer region of rDNA. Despite initial amphotericin B and voriconazole therapy, the child's jaundice subsided and he died 7 months later. In addition to pathogenic aspects of Exophiala dermatitidis, the diagnostic approaches and relevant therapeutic strategies are discussed. PMID- 19184770 TI - Host immune response against Scedosporium species. AB - Scedosporium apiospermum and Scedosporium prolificans cause therapy-refractory infections in immunocompromised and immunocompetent hosts. While innate immune response is believed to be critical for the host defense against these fungi, its role has only recently been elucidated. Undefined pathogen-associated molecular patterns on the surface of conidia and hyphae are recognized by pattern recognition receptors (PRRs) on the membrane of phagocytes, and the signal is transmitted intracellularly. PRRs that are important in the recognition of both fungal species are human Toll-like receptors (or Toll receptors in Drosophila melanogaster) and dectin-1. These induce signals responsible for the activation of genes leading to an effective host defense, especially those encoding pro inflammatory cytokines. Both species are efficiently phagocytosed and elicit an oxidative burst by neutrophils and monocytes. While cytokines, such as interleukin-15, granulocyte-macrophage colony-stimulating factor, granulocyte colony-stimulating factor and interferon-gamma, have been found in vitro to variably modulate antifungal activity of human phagocytes, cytokines in vivo activities are less well documented. Certain antifungal agents exert immunopharmacological effects on phagocytes against S. apiospermum and S. prolificans. Translation of these in vitro findings to appropriate in vivo systems and into clinical trials may lead to improved strategies for augmenting innate host defenses in patients infected with these emerging pathogens. PMID- 19184771 TI - Persistence of Candida species in the respiratory tract of cystic fibrosis patients. AB - It is still controversial as to whether Candida spp. are transient or persistent colonizers of the respiratory tract of cystic fibrosis (CF) patients. We conducted a prospective study of 56 CF patients over a 30 month period to assess the distribution and persistence of different Candida spp. In vitro antifungal susceptibility testing was performed and the C. albicans isolates were typed with CARE-2 hybridization and other Candida spp. by RAPD-PCR for persistence and transmission. We found that the mean persistence of the most frequent Candida spp. was >or= 9 months. In patients from whom more than 10 isolates were recovered, we noted that at least 30% were genetically related and transmission of C. albicans in siblings was observed. The majority of all isolates were susceptible to all antifungals tested. We concluded that there was long-term persistence of Candida in the respiratory tract of CF patients and that transmission between siblings may be one possible means of acquisition. Whether long-term colonization with Candida strains can contribute to the chronic infection and inflammation in the CF lung requires further investigation. PMID- 19184772 TI - Heterogeneity of proteins expressed by Brazilian Sporothrix schenckii isolates. AB - The profiles of proteins present in the exoantigens of Brazilian Sporothrix schenckii isolates were studied and compared by electrophoresis (SDS-PAGE). Thirteen isolates from five different regions of Brazil (1,000 to 2,000 km apart) and ten from a more limited region (200 to 400 km apart within the state of Sao Paulo) were cultured in Sabouraud, M199 and minimum (MM) media. Qualitative and quantitative differences in the expression of proteins, which varied according to the medium and the isolate, were observed. Fractions with the same MW but varying in intensity were detected, as well as fractions present in 1 isolate but absent in others. Dendrograms were constructed and isolates grouped based on the fractions obtained, irrespective of the intensity. The results showed that Brazilian S. schenckii isolates express different protein profiles, a feature also present in isolates from a more restricted region. The exoantigens were found to have a maximum of 15 protein fractions, ranging in MW from 19-220 KDaltons depending on the medium used for the cultures. These data show the great heterogeneity of Brazilian S. schenckii protein expression. PMID- 19184774 TI - Oxidative stress response of Inonotus obliquus induced by hydrogen peroxide. AB - While the medicinal fungus Inonotus obliquus produces polyphenols as one of its main metabolites in natural habitats, it accumulates less polyphenols under laboratory conditions. In this study we found that the continuous addition of 1 mM H(2)O(2) at a rate of 1.6 ml/h into a submerged culture of the fungus enhanced its production of mycelia, melanins, flavonoids and hispidin analogs (HA). Simultaneous exposure of the fungus to both H(2)O(2) and arbutin resulted in reduced production of mycelia, glycosylated flavonoids (GF) and HA, and inhibition of melanogenesis. However, superoxide dismutases (SOD) and catalase (CAT) activity were enhanced following the addition of H(2)O(2) or H(2)O(2) plus arbutin. The maximum levels of SOD and CAT activities reached 355.2 U/mg protein and 39.8 U/mg protein respectively in H(2)O(2)-added medium, and 264 U/mg protein and 35.9 U/mg protein respectively in H(2)O(2) plus arbutin medium. Thus, detoxification of H(2)O(2) is conducted mainly by polyphenols under normal physiological conditions, and by both polyphenols and antioxidant enzymes under oxidative stress when melanogenesis is inhibited. Although enhanced HA production occurred after melanogenesis inactivation, total extracellular polyphenol levels were reduced. These findings suggest that enzymatic activities convert superoxide to H(2)O(2), and non-enzymatic mechanisms are largely responsible for detoxifying H(2)O(2). Enhanced production of melanins is the most important non-enzymatic response of this fungus against oxidative stress. PMID- 19184775 TI - Utility of real-time PCR for the detection of Paracoccidioides brasiliensis DNA in the diagnosis of imported paracoccidioidomycosis. AB - An increase in immigration from endemic regions has resulted in a number of cases of paracoccidioidomycosis (PCM) being imported into Spain. A molecular diagnostic technique based on real-time PCR was developed for the detection of Paracoccidioides brasiliensis DNA in both culture and patients' clinical samples. A Molecular Beacon probe was used, labelled with FAM and directed at the ITS1 region of ribosomic DNA. The detection limit of the technique developed was 1 fg of fungal DNA per microl of sample. This procedure proved to be very reproducible and specific. The technique was tested with cultures of 12 clinical strains and on samples from two patients with proven PCM. Real-time PCR was positive for all the culture strains, as well as those from both patients. By samples, the technique was positive in sputum and tissue biopsies but less useful on blood samples. Samples were analyzed several months after patient treatment, detecting a small amount of fungal DNA in one respiratory sample. This technique of real time PCR is a sensitive method for rapid diagnosis of paracoccidioidomycosis and could serve to monitor patients after treatment has begun. PMID- 19184776 TI - Glycocalyx volume: a critical review of tracer dilution methods for its measurement. AB - A clinical measure of endothelial glycocalyx structure would have great potential importance, because lesions of the glycocalyx may be the first changes to occur in diabetes and in a wide range of vascular diseases. A method recently described by Nieuwdorp et al. for estimating the volume of the luminal glycocalyx of the entire human vascular system would seem to be the first attempt to develop a measure of this kind. It is based on the tracer dilution principle, and this review considers the principles and conditions that underlie this method and the extent to which the conditions appear to have been fulfilled in this case. Our analysis raises two questions about 1) the estimation of the concentration of the tracer (dextran 40) at zero time and 2) the estimation of plasma volume, both of which can be answered by changes in experimental protocol. A third question, concerning the partition coefficient of the tracer between plasma and the fluid within the glycocalyx, cannot be answered at the present time, and until it has been resolved, glycocalyx volume cannot be estimated from the dilution of a macromolecular tracer. PMID- 19184777 TI - The vasodilatory response of skin microcirculation to local heating is subject to desensitization. AB - BACKGROUND: In humans, local heating increases skin perfusion by mechanisms dependent on nitric oxide (NO). Because the vascular effects of NO may be subject to desensitization, we examined whether a first local thermal stimulus would attenuate the hyperemic response to a second one applied later. METHODS: Twelve healthy young men were studied. Skin blood flow (SkBF) was measured on forearm skin with laser Doppler imaging. Local thermal stimuli (temperature step from 34 to 41 degrees C maintained for 30 minutes) were applied with temperature controlled chambers. We also tested the influence of prior local heating on the vasodilation induced by sodium nitroprusside (SNP), a donor of NO. RESULTS: On reheating the same spot after two hours, the response of SkBF (i.e., plateau SkBF at 30 minutes minus SkBF at 34 degrees C) was lower than during the first stimulation (mean+/-SD 404+/-212 perfusion units [PU] vs. 635+/-100 PU; P<0.001). There was no such difference when reheating after four hours (654+/-153 vs. 645+/ 103 PU; P=NS). Two, but not four, hours after local heating, the response of SkBF to SNP was reduced. CONCLUSION: The NO-dependent hyperemic response induced by local heating in human skin is subject to desensitization. At least one part of the mechanism implicated consists of a desensitization to the effects of NO itself. PMID- 19184778 TI - Neuropsychological functioning in children with non-syndromic cleft of the lip and/or palate. AB - OBJECTIVE: Evaluate neuropsychological functioning in children with non-syndromic cleft of the lip and/or palate (NSCL/P) through profile variance within type of cleft and comparisons to controls. METHODS: Children ages 7 to 17 years participated; 66 had a diagnosis of NSCL/P and 87 were healthy controls. Neuropsychological tests of language, visual-perceptual, executive functioning, and memory skills were administered. Between- and within-group differences were assessed. RESULTS: Within cleft types, children with NSCLP had an even profile with equal Verbal and Performance IQ (VIQ and PIQ, respectively). Children with non-syndromic cleft palate only (NSCP) had significantly lower VIQ than PIQ, while children with non-syndromic cleft lip only (NSCL) showed a nonsignificant trend for higher VIQ than PIQ. Overall, subjects with NSCL/P performed lower on measures of expressive language and verbal memory than controls. CONCLUSIONS: While deficits in verbal and memory skills for children with NSCL/P remain apparent, there is still uncertainty around the possible influence of cleft type on the pattern of deficits. PMID- 19184779 TI - Intra-individual variability among children with ADHD on a working memory task: an ex-Gaussian approach. AB - Intra-individual variability on a computer-based working memory task was examined among 25 children/adolescents with ADHD and 24 typically developing peers. Participants completed the Visual Serial Addition Task (VSAT) and reaction time data were fit to an ex-Gaussian distribution. ADHD participants demonstrated significantly more variable performance than controls, and effects of working memory load were observed. Event rate, however, had no influence on group differences in performance. Follow-up correlations revealed associations between VSAT performance and ADHD symptomatology. This study supports intra-individual variability as a hallmark feature of ADHD beyond the domain of response inhibition and reinforces the need to consider variability in ADHD more broadly. PMID- 19184781 TI - Editorial. PMID- 19184780 TI - Advancing the profession of clinical neuropsychology with appropriate outcome studies and demonstrated clinical skills. AB - Healthcare economics and politics require clinical neuropsychologists to clearly document the value of their services. Therefore, advancing clinical neuropsychology as an economically viable profession requires carefully designed outcome studies and clear demonstration of the value of neuropsychologists' judgment in various patient care matters. In this paper we propose steps for achieving these two goals. PMID- 19184782 TI - Iron storage diseases in birds. AB - Parenteral iron is toxic to many species but, because the uptake of iron from the diet is regulated in the intestine, acute intoxication is not seen under natural conditions. Chronic ingestion of large amounts of absorbable iron in the diet can lead to the storage of iron in the liver in many species, including humans. The excess iron is stored within hepatocytes as haemosiderin and can be quantitatively assessed by liver biopsy or at necropsy using special stains such as Perls iron stain and/or biochemical tests. Iron may also be found within the Kupffer cells in the liver and the macrophage cells of the spleen especially where concurrent diseases are present such as haemolytic anaemia, septicaemia, neoplasia and starvation. Iron accumulation in the liver, also known as haemosiderosis, may not always be associated with clinical disease although in severe cases hepatic damage may occur. It is probable that concurrent disease conditions are largely responsible for the degree and nature of the pathological changes described in most cases of haemosiderosis. In some human individuals there may be a genetic predisposition to iron storage disease, haemochromatosis, associated with poor regulation of iron uptake across the intestine. In severe cases iron pigment will be found in the liver, spleen, gut wall, kidney and heart with subsequent development of ascites, heart failure and multisystem pathology. Clinical disease associated with accumulation of iron in the liver, and other tissues, has been reported in many species of bird although it is most commonly reported in Indian hill mynas ( Gracula religiosa ) and toucans ( Ramphastos sp ). It is likely that the tolerance to the build up of tissue iron varies in individual species of bird and that the predominant predisposing factors may differ, even within closely related taxonomic groups. PMID- 19184783 TI - Transfer of antibodies elicited by baculovirus-derived VP2 of a very virulent bursal disease virus strain to progeny of commercial breeder chickens. AB - A baculovirus-derived recombinant VP2 (rVP2) subunit vaccine elicited anti infectious bursal disease virus (IBDV) antibodies in commercial flocks. The induced antibody levels were similar to those evoked against IBDV by a commercial vaccine. The levels remained higher than that of the negative controls for at least four and a half months in commercial chickens. The antibodies were also transferred to their offspring and were detected in the blood of the progeny for at least 20 days after hatching. These results, along with former data, that show that antibodies elicited by baculovirus rVP2 confer protection to chickens from IBDV [J. Pitcovski et al. (1996), Insect cell-derived VP2 of infectious bursal disease confers protection against the disease in chickens. Avian Diseases, 40, 753-761], imply that the baculovirus-derived rVP2 subunit may serve as a successful vaccine for commercial breeding flocks. PMID- 19184784 TI - Contamination of Marek's disease vaccine suspensions with Enterococcus faecalis and its possible role in amyloid arthropathy. AB - Samples of hatchery air (hatcher and processing room), Marek's disease vaccine suspensions and injection needles collected during chick processing, revealed variable levels (< 500 to 10(6) colony forming units (cfu)/m(3) air, < 10 to 10(6) cfu/ml vaccine suspension, and 9500 to 61000 cfu/needle) of Enterococcus faecalis contamination. This observation suggests a possible infection route in 1 day-old chickens through intramuscular vaccination of Marek's disease vaccine contaminated with arthropathic and amyloidogenic E. faecalis, which could lead to amyloid arthropathy. Pulsed-field gel electrophoresis (PFGE) DNA restriction endonuclease fragment analysis of E. faecalis strains obtained from two hatcheries revealed a predominant PFGE pattern in one hatchery, while one isolate with an almost identical PFGE pattern to an amyloid arthropathy inducing isolate was found. PMID- 19184785 TI - Oral Newcastle disease vaccination trials in Ethiopia. AB - Vaccination experiments were carried out in Ethiopia to study the efficacy of the NDV-I(2) vaccine against challenge with an Ethiopian velogenic strain of NDV. In experiment A, which comprised 300 broiler chicks, the efficacy of the ocular/drinking water application of the HB1/La Sota vaccine was compared with the ocular/drinking water and the feed application of the NDV-I(2) vaccine on untreated barley and sorghum. The NDV-I(2) vaccine applied by eye-drop or drinking-water protected the chickens against challenge as efficiently as combined HB1/La Sota vaccination but untreated barley and sorghum were unsuitable vaccine carriers. The vaccine virus could not be recovered and chickens neither seroconverted nor were they protected. In experiment B, 120 broiler chicks were divided into 6 treatment groups. One group each received NDV-I(2) vaccine mixed with untreated barley or sorghum which was applied immediately, or 14h after mixing and standing at ambient temperature. The fifth group was vaccinated intraocularly and via the drinking water with the NDV-I(2) vaccine. The sixth group remained untreated. Experiment B confirmed the results of experiment A. In experiment C, 100 chicks were divided into 5 groups of 20 chickens each. One group each received the NDV-I(2) vaccine on parboiled barley or sorghum as vaccine carriers 0 and 6h after mixing. The last group remained untreated. Parboiled barley given 0 or 6 h and parboiled sorghum given 0 h after mixing with the vaccine led to seroconversion and protection of the chickens. Parboiled sorghum given 6h after mixing with the vaccine did not. It is concluded that the thermostable NDV-I(2) vaccine may be a suitable vaccine for oral application under Ethiopian conditions. PMID- 19184786 TI - Diagnosis of turkey meningoencephalitis virus infection in field cases by RT-PCR compared to virus isolation in embryonated eggs and suckling mice. AB - Turkey meningoencephalitis (TME) is a paralytic epornitic disease of turkeys caused by turkey meningoencephalitis virus (TMEV), an arthopod-borne flavivirus belonging to the Ntaya serogroup VI. A TMEV specific RT-PCR was compared with classical techniques for TMEV diagnosis, which include virus isolation in 8-day old chicken embryonated eggs and suckling mice, on 17 TME flocks with neurological signs that occurred during the fall of 1997. In 11/17 flocks both the RT-PCR and the virus isolation methods detected virus, in 4/17 flocks a negative diagnosis was obtained by both methods, and two flocks were positive by RT-PCR only. In four flocks RT-PCR only detected virus after inoculation into embryonated eggs or suckling mice. There was a dose response effect in the yield of the RT-PCR product. Direct examination of turkey brains yielded bands of low to medium intensity. Use of RT-PCR after embryo and/or mouse inoculation resulted in products of far greater intensity. Thus, RT-PCR can be successfully used to amplify TMEV RNA in the brains of diseased turkeys but a negative result would require egg and mouse inoculation for enrichment of virus prior to RT-PCR. PMID- 19184787 TI - Pseudomonas aeruginosa infections in kori bustards (Ardeotis kori). AB - Pseudomonas aeruginosa associated upper respiratory tract infections occurred in a flock of 19 juvenile kori bustards. Morbidity was 32% and all birds recovered after parenteral treatment with combinations of enrofloxacin, gentamicin and piperacillin, although gentamicin toxicity was suspected in one bird. A marked haemoresponse occurred 3-10 days after the onset of clinical signs and included leukocytosis, heterophilia, monocytosis and thrombocytosis. Predisposing factors to infection included recent translocation, changes to flock size, extreme temperatures and age. PMID- 19184788 TI - Phylogenetic analysis of parvoviruses isolated in Taiwan from ducks and geese. AB - Two major outbreaks of parvovirus infection occurred in domestic waterfowls in Taiwan in the last two decades; the first was in 1982 and the second in 1989/1990. Parvoviruses isolated in the two outbreaks were sequenced between nucleotides 142 and 680 of the VP3 gene. Sequence comparisons reveal that these viruses could be divided into two groups respectively related to goose parvovirus (GPV) and Muscovy duck parvovirus (MDPV). Nucleotide differences between ''GPV'' and ''MDPV'' groups range from 16.2% to 19.4%. In comparison, the differences within the ''GPV'' group are only 0-6.5%, while those within the ''MDPV'' group are only 0.2-1.7%. Phylogenetic analysis reveals that parvoviruses isolated in the 1982 outbreak in Taiwan are all GPV-related, whereas those isolated in the 1989/1990 outbreak are all MDPV-related. GPV-related isolates from Taiwan were separated into two groups, Thai group and European group. In comparison, all MDPV related isolates from Taiwan are clustered in a single group that is closely related to a French MDPV isolate. The MDPV-related infection in Taiwan occurred at almost the same time in 1989 as the MDPV outbreak in France. The close phylogenetic relationship between the ''MDPV'' Taiwanese isolates and the French MDPV isolate exhibited on the VP3 fragment investigated suggests that they should be compared more deeply, to look for a possible common origin. The MDPV-related 1985 isolate might be a candidate. PMID- 19184789 TI - Central nervous disease and blindness in Nankeen kestrels (Falco cenchroides) due to a novel Leucocytozoon-like infection. AB - The histopathological and electronmicroscopic features of a novel parasitic infection causing seasonal epizootics of central nervous disease in Nankeen kestrels (Falco cenchroides) from Western Australia is described. Thorough necropsy, haematological and transmission electronmicroscopical examinations were performed on thirteen affected kestrels between 1996 and 1999. All had an extensive vasculocentric granulomatous meningoencephalomyelitis, pectenitis and proliferative arteritis with endothelial parasitic cysts measuring 40 to 60 mu m in diameter. Arterioles in the brain, optic papillae, pecten and kidney were most consistently affected. In some birds there was severe haemorrhage from the pecten. Less commonly, parasitic cysts were seen in the arterioles of the lung, liver, heart and intestines. Endothelial schizonts contained many spherical merozoites measuring 1 mu m in diameter that were also present free within the lumen of affected arterioles. Haematological examination in three of seven affected kestrels tested demonstrated rare, basophilic intracytoplasmic inclusions in leucocytes with distorted nuclei consistent with Leucocytozoon-like gametocytes. Day-old chickens inoculated with brain homogenate failed to develop similar lesions. PMID- 19184790 TI - RFLP analysis of recent Northern Ireland isolates of infectious laryngotracheitis virus: comparison with vaccine virus and field isolates from England, Scotland and the Republic of Ireland. AB - Restriction fragment length polymorphism (RFLP) analysis was used to assist epidemiological investigations following the recent introduction of infectious laryngotracheitis virus (ILTV) to commercial poultry flocks in Northern Ireland (NI). A 4.9 kbp PCR product of the ILTV ICP4 gene was generated from each of 16 field isolates of ILTV originating from England, Scotland, NI and the Republic of Ireland (RoI) and of the single vaccine strain currently licenced for use within the United Kingdom. With the exception of isolate PV6/94 from RoI, all field isolates generated RFLP patterns, following digestion with HaeIII, similar to that obtained using the vaccinal strain. Following MspI digestion, NI isolates were indistinguishable from the vaccinal strain and recent English isolates. However, one English and one Scottish isolate, both made prior to the introduction of vaccination, and two isolates from RoI generated a second pattern following digestion with MspI. PMID- 19184791 TI - Houghton trust international travel and subsistence grants. PMID- 19184792 TI - Editorial. PMID- 19184793 TI - Detection of infectious bronchitis virus. AB - The detection methods for infectious bronchitis virus (IBV) are reviewed. Advantages and disadvantages of available techniques of IBV detection by virus isolation, antigen or genome detection, and serology are discussed. Factors of influence on the level of success in detection of IBV after a disease outbreak are discussed, as are the possibilities and dangers of strain classification by protectotyping, serotyping, epitope-typing and genotyping. PMID- 19184794 TI - Newcastle disease in ostriches (Struthio camelus) - a review. AB - Newcastle disease (ND) in ostriches was first reported in zoo birds in the 1950s. An outbreak of ND in commercial ostriches in Israel was reported in 1989, but of more significance were the ND infections of ostriches in southern Africa during the 1990s as a result of assumed spread from commercial domestic fowl. The latter outbreaks were of particular concern due to the boom in international trade in ostriches and their products at that time. ND virus appeared to spread only slowly through affected ostrich flocks, and the clinical signs and mortality seen varied considerably with age. In field and experimental infections, morbidity, mainly in the form of nervous signs, and mortality could be extremely high in young birds but low or absent in adults. Characterization of ND viruses isolated during outbreaks in ostriches has shown them to be indistinguishable from viruses infecting chickens in the locality. Experimental challenge has shown ND poultry vaccines to be protective in ostriches, and several workers have proposed vaccine regimens employing both live and inactivated vaccines, usually given more frequently and employing much higher doses than recommended for chickens. Indirect and blocking enzyme-linked immunosorbent assay (ELISA) tests have been developed for the detection of ND antibodies in ostrich sera. Although some reports have suggested problems using the haemagglutination inhibition(HI)tests, others have recorded good correlation between ELISA, HI and virus neutralization tests. Because of their size and, as a consequence, the method of rearing, ostriches are quite different to conventional poultry and respond differently, in terms of disease and spread, when infected with ND virus. However, at present, for the international control of ND, there would appear to be little alternative to treating ostriches as poultry for the purposes of trade. PMID- 19184795 TI - The use of conventional radiography and computer-assisted tomography as instruments for demonstration of gross pathological lesions in the cranium and cerebrum in the crested breed of the domestic duck (Anas platyrhynchos f.dom.). AB - According to literature reports, the crested breed of the domestic duck (Anas platyrhynchos f. dom.) is described as a variety with high pre- and post-natal mortalities, malformations in skull and brain anatomy, and various central nervous deficiencies in affected birds. A detailed examination as to the cause and pathogenesis of these neurologic disorders has not yet been conducted, nor has an explanation concerning the inheritance of the feather crest been found. By using two non-invasive radiographic techniques, conventional radiography and computer-assisted tomography, living ducks were examined. Our studies have demonstrated that brain and skull anatomy of ducks of various breeds (Crested, Abacot Ranger, and German Pekin) can be well visualized using computer-assisted tomography. The Crested ducks investigated in this study showed abnormal fatty tissue deposits in the tentorium cerebelli, cranial malformations, and variable bone formations in the thickened hypodermis of the crest. In comparison with computer-assisted tomography, only parts of the skull changes were detected with conventional radiography; in no case could fat bodies be seen with this imaging method. An increase of the cranial capacity in the Crested ducks compared with other breeds examined (Abacot Ranger, German Pekin ducks) was found in morphometric studies carried out by computerassisted tomography. This increase of cranial capacity in Crested ducks results from an increase of the tentorial fatty depot during craniogenesis, as an enlargement of the fat body can influence cranium growth as long as the cranial bones are not yet fused. Thus, in comparison with other phenotypically similar domesticated birds like the crested chicken, the expression of the feather crest in domestic ducks demonstrates symptomatologic differences. PMID- 19184796 TI - Assessment of liver function in chickens using galactose and indocyanine green clearances. AB - Values for galactose and indocyanine green (ICG) clearances, and plasma and serum biochemical markers of liver dysfunction were determined in normal chickens and following coeliotomy, and compared with birds after partial hepatectomy. Clearance tests, and serum and plasma biochemistry were performed 4h, and 4 and 7 days after surgery. Coeliotomy and manipulation of the liver did not delay clearance of either compound. Partial hepatectomy resulted in elevation of galactose single point concentrations but did not significantly alter galactose clearance (GEC) values. Clearance values of ICG were not significantly altered. Biochemical values were not significantly elevated in birds after a partial hepatectomy in comparison with birds after coeliotomy. Galactose single point concentrations have the potential to become a simple, relatively non-invasive method of screening for liver disease, with GEC tests having the potential to quantify the degree of loss of functional hepatic mass. PMID- 19184797 TI - Longevity of augmented phagocytic activity of heterophils in neonatal chickens following administration of Salmonella enteritidis-immune lymphokines to chickens. AB - Previously, our laboratory reported an existing relationship between ontogenesis of heterophil activity and susceptibility to Salmonella enteritidis infection in poultry during the first week post-hatch. The prophylactic administration of S. enteritidis-immune lymphokines has been shown to enhance heterophil function in vitro and reduce S. enteritidis organ invasion in 1-day-old chicks. However, how long the heterophils remain activated is currently unknown. The objective of this research was to evaluate the duration of enhanced heterophil phagocytosis of S. enteritidis following the prophylactic administration of a single injection of S. enteritidis-immune lymphokines to neonatal chicks. Administration of S. enteritidis-immune lymphokines on the day of hatch resulted in a significant increase in heterophil phagocytic activity (P < 0.05) through day 5 post-hatch. No significant differences in phagocytic activities of heterophils from control and S. enteritidis-immune lymphokinetreated chicks was demonstrated from day 6 to 14 post-hatch. These data suggest that the administration of S. enteritidis immune lymphokines on day of hatch potentiates heterophil phagocytic activity during a critical period of susceptibility to Salmonella infection in neonatal chicks. The immune lymphokine-induced enhancement of heterophil activity subsides by day 5 as the chick's natural defenses mature and are able to resist infections without exogenous augmentation. PMID- 19184798 TI - Thermophilic Campylobacter spp. in Danish broiler production: a cross-sectional survey and a retrospective analysis of risk factors for occurrence in broiler flocks. AB - In order to elucidate the rate of thermophilic Campylobacter spp. carriage in Danish broiler production and to identify risk factors for occurrence of campylobacter in broiler flocks, a total of 88 randomly selected broiler flocks were tested for campylobacter infection, and a subsequent study of risk factors based on a questionnaire was conducted. The sample material comprised cloacal swabs from live birds before slaughter, and neck skin samples from carcasses at the end of the processing line. A total of 52% of the flocks were found Campylobacter spp.-positive before slaughter. At the end of processing, 24% of the flocks were positive. The species distribution was 87% Campylobacter jejuni, 8% Campylobacter coli and 5% Campylobacter lari. The following parameters were identified as significant risk factors: lack of a hygiene barrier (odds ratio (OR) = 3.1, 1.1 < OR < 9.3), presence of animals in the vicinity of the broiler house on farms with a missing hygiene barrier (OR = 7.0, 1.6 < OR < 33.9), livestock other than chickens on farms with a missing hygiene barrier (OR = 7.6, 1.4 < OR < 44.9), dividing the flock into batches for staggered slaughter (OR = 6.8, 1.2 < OR < 49.3), a down period of less than 14 days (OR = 5.0, 1.2 < OR < 22.6), and feeding purchased wheat rather than home-grown wheat (OR = 3.1, 1.0 0.05) on replication efficiency between these two serotypes was found based on the results of total RNA concentration and RT-PCR results of 10-fold dilution of RNA at the indicated time points. By inoculating heat-inactivated Mass 41 with live JMK into embryos, only JMK was detected by RT-PCR-RFLP and no dominating interference was observed. Data suggest that the dominance is not due to the replication efficiency of Mass 41 and JMK; a receptor-mediated mechanism might be responsible for it. PMID- 19184837 TI - Re-emerging fowlpox: evaluation of isolates from vaccinated flocks. AB - Vaccines of fowlpox or pigeonpox virus origin have been routinely used for more than half a century to prevent fowlpox in commercial poultry in areas where the disease is endemic. However, in recent years, outbreaks of fowlpox have occurred in previously vaccinated flocks. One possible explanation for this problem is the emergence of variant strains of fowlpox virus (FPV). A second, not mutually exclusive, postulate is that the novel FPV exhibit enhanced virulence due to the integration of avian reticuloendotheliosis virus (REV) into their genomes. To determine if immunological variance and/or the acquisition of REV nucleotide sequences could be responsible for the ineffectiveness of current vaccines, the ability of two commercial vaccine viruses and four, recently isolated, field strains to protect chickens against challenge with one of the more virulent field viruses was evaluated. Adequate protection was provided by the vaccines and two of the four field isolates. Interestingly, the two isolates that were not protective, as well as the challenge strain, failed to elicit a strong humoral antibody response. As to possible REV participation, an antibody response to this virus was only found in those chickens receiving one of the ''protective'' field strains, despite the presence of REV coding sequences in all four field viruses. While REV long terminal repeats of variable lengths were detected in the genomes of all FPV strains used in this study, only the DNAs of the field strains appeared to have intact REV provirus. This retention of foreign DNA may enhance the pathogenesis of FPV, although other factors may be involved. PMID- 19184838 TI - Molecular epidemiology of Salmonella gallinarum in chickens in Tanzania. AB - A molecular epidemiological investigation of Salmonella gallinarum infection in scavenging local chickens and commercial layers in Tanzania was conducted between August 1997 and April 1998. A total of 1152 chickens were randomly selected from 10 villages and seven commercial farms. For serological and cultural prevalence studies, 1152 blood samples and 912 cloacal swabs were collected. In scavenging local chickens, the individual serological and cultural prevalences were 6.3 and 0%, while the prevalences were significantly higher in commercial layers at 18.4 and 2.6% (P < 0.001), respectively. The risk of infection in flocks of scavenging local chickens that had contact with commercial chickens was six times greater than the risk of infection in flocks of scavenging local chickens that had no contact with commercial chickens. Thirty-four S. gallinarum isolated from commercial chickens in this study, together with 29 Tanzanian historical isolates, were characterized using plasmid profiling and ribotyping. Fifty-one isolates contained both 85 and 2.5kb plasmids, five isolates contained only one plasmid of 85kb, and seven isolates had no plasmids. Ribotyping using HindIII restriction endonuclease demonstrated seven different ribotypes. Forty-seven isolates had similar results in both typing systems, suggesting they belonged to one clone. It is concluded that S. gallinarum infection in chickens in Tanzania is more prevalent in commercial layers than in scavenging local chickens. One strain of S. gallinarum from chickens first isolated in a Dar es Salaam hatchery was found to be common throughout the country. PMID- 19184839 TI - The effect of vaccination on the course of an experimental Salmonella typhimurium infection in racing pigeons. AB - Two killed adjuvanted vaccines were evaluated for their efficacy against salmonellosis in pigeons. Both vaccines contained whole-cell formaldehyde inactivated Salmonella typhimurium var. Copenhagen bacteria. Two groups of 10 pigeons were inoculated twice with a 3-week interval with one of the vaccines. Ten weeks after the second vaccination, all vaccinated pigeons as well as 10 non vaccinated birds were challenged by crop inoculation of 10(9) colony-forming units of a Salmonella serotype Typhimurium var. Copenhagen strain. Ten pigeons that were not vaccinated or challenged served as negative controls. As determined by clinical examination, plasma chemistry and necropsy, neither vaccine induced protection against challenge. The only significant effects observed were a reduction in the level of faecal shedding and a less severe polydipsia in the pigeons treated with one of the two vaccines. Results of this study indicate that vaccination against salmonellosis in pigeons with killed vaccines may not be very useful. However, the ability of certain vaccines to reduce shedding may contribute to the control of salmonellosis in infected pigeon lofts. PMID- 19184840 TI - Characterization of Haemophilus paragallinarum isolates from Mexico. AB - The carbohydrate fermentation, antimicrobial drug resistance and serological properties of 40 isolates of Haemophilus paragallinarum from outbreaks of infectious coryza in Mexico are described. Four biochemical biovariants and five antimicrobial drug resistance patterns were recognized. All isolates were serotyped by the Page scheme, with 21 isolates being assigned to serogroup A, five isolates to serogroup B and 14 isolates to serogroup C. PMID- 19184841 TI - Bacterial chondronecrosis with osteomyelitis ('femoral head necrosis') of broiler chickens: a review. AB - Bacterial chondronecrosis with osteomyelitis (BCO) in chickens was first reported in 1972 and is now recognized as an important cause of lameness in broiler chickens. Recent systematic studies of causes of lameness in birds reared in Northern Ireland have shown that it was the most common cause of lameness, being present in 17.3% of lame birds. Furthermore, it was also detected in birds presented as ''found dead''. Overall losses in male birds due to BCO were estimated to be 0.75% of all birds placed, which, in addition to welfare concerns, represents considerable economic loss. The disease has been seen in birds ranging from 14 to 70 days of age, but most cases occurred around 35 days old. It is most commonly caused by Staphylococcus aureus, but Escherichia coli, coagulase-negative staphylococci and Enterococcus spp. are sometimes involved, as are, rarely, other bacteria. The lesions are most commonly found associated with the growth plates of long bones, particularly the proximal growth plate of the femur and tibiotarsus, but other bones may also be affected. Since lesions were visible to the naked eye in only 40 to 67% of cases, histological examination is recommended where no lesions are visible macroscopically.As the lesion may be present in only one growth plate, and because histological examination is often not carried out, BCO is almost certainly underdiagnosed. The exact pathogenesis of the condition is unknown, but it is thought that adherence of blood-borne bacteria to exposed cartilage at the tips of metaphyseal blood vessels is fundamental. Under controlled experimental conditions, infection of birds with the immunosuppressive viruses chicken anaemia virus and infectious bursal disease virus increased the incidence of the disease, while restricting feed intake reduced the incidence of disease. S. aureus strains identical to, or closely related to, isolates recovered from naturally occurring cases of the disease (as determined by pulsed-field gel electrophoresis) have been recovered from fluff debris in hatcheries, and also from the environment of breeding flocks, indicating that infection in the breeding farm and in the hatchery could be an important source of infection. It has also been shown that humans can carry poultry strains of S. aureus on their hands. There is a higher incidence of BCO in birds hatched from floor eggs. Thus, hygiene and management practice on breeder farms and in the hatchery may influence the occurrence of the disease. Bacteraemia is a prerequisite for BCO. Indeed, in some flocks suffering losses due to BCO, there are also losses due to staphylococcal septicaemia. Thus, appropriate treatment of affected flocks should reduce losses due to septicaemia. It should also reduce the occurrence of bacteraemia and the development of further cases of BCO. However, birds in which BCO has already developed, are unlikely to respond to treatment. Control of BCO by vaccination seems unlikely in the short term. Simple bacterins have not been effective and much basic research is needed to identify the important virulence factors. Furthermore, more than one type of bacterium is capable of causing the disease. Bacterial interference has been used successfully in humans and turkeys to prevent staphylococcal diseases, and warrants investigation for the prevention of BCO in chickens. This may have an advantage in that the interfering bacterium may also exclude some of the other bacteria that can cause BCO. The recent development of a disease model in which S. aureus is given by a natural route allows the potential for further investigation of the role of predisposing factors, and intervention strategies, including vaccination and bacterial interference, for the prevention of BCO. PMID- 19184842 TI - Isolation of acutely transforming subgroup J avian leukosis viruses that induce erythroblastosis and myelocytomatosis. AB - Avian leukosis virus of subgroup J (ALV-J), isolated in the late 1980s, predominantly causes myelocytic myeloid leukosis in meat-type chickens. In the past few years, we have observed the occurrence of lesions indicative of erythroblastosis in ALV-J-infected flocks and, in this paper, we report the isolation of ALV-J strains from such flocks. Three of these isolates were acutely transforming viruses, as shown by their ability to transform bone marrow cell cultures. The bone marrow cultures transformed by these virus isolates were very similar to the myeloid cells transformed by the ALV-J strain 966. However, the infection of meat-type chickens with these isolates either as embryos or as 1-day old chicks resulted in the induction of erythroblastosis as well as myelocytomatosis. Other histopathological changes observed in the inoculated birds included neoplastic lesions such as cholangioma and testicular cell tumour, and non-neoplastic lesions such as lymphomyeloid hyperplasia. This report demonstrates that highly oncogenic ALV-J, capable of inducing a different spectrum of disease other than the widely reported myelocytomatosis, could be established in naturally infected flocks. PMID- 19184845 TI - WVPA Matters. PMID- 19184844 TI - International meetings. PMID- 19184846 TI - Introducing Commentary articles. PMID- 19184847 TI - Physiological, management and environmental triggers of the ascites syndrome: a review. AB - In meat-type chickens, an inadequacy of vascular capacity for blood flow through the lung to provide the tissues with the oxygen needed for rapid growth is the primary cause of pulmonary hypertensioninduced ascites. There are a variety of other factors that can trigger the ascites syndrome. These factors may cause increased blood flow because of a higher metabolic rate (cold, heat, certain nutrients, chemicals, etc.) or they may cause pulmonary hypertension-induced ascites in rapidly growing chickens because of greater resistance to blood flow in the lung by: (i) increased blood viscosity or red blood cell rigidity; or (ii) reduced vascular capacity in the lung. Some secondary factors, such as high sodium from salt in feed or water, may cause both increased flow and increased resistance to flow. Measures to reduce the ascites syndrome must address the primary genetic cause of insufficient vascular flow capacity in the lung and oxygen delivery to tissues, and the secondary factors that increase oxygen requirement, blood flow and the resistance to blood flow in the lung. PMID- 19184848 TI - Isolation and identification of avian leukosis viruses: a review. AB - Avian leukosis virus (ALV) is the most common naturally occurring avian retrovirus that can cause a variety of neoplastic disease conditions in chickens. In addition to causing neoplasia, ALV is known to be associated with reduced productivity and other production problems in affected flocks. Biological and molecular assays for the detection of ALV are very useful in identification and classification of new isolates, safety testing of vaccines and in testing pathogen-free and other breeder flocks for freedom of virus infection. However, such assays are not particularly helpful in the diagnosis of virus-induced neoplastic disease of poultry, as avian oncogenic viruses are widespread and infection in the absence of tumor formation is common. Current technology based on molecular and antigenic characteristics of the virus is being used to develop more sensitive and specific procedures for isolation and identification of ALV. This review is primarily focused on the discussion of current technology most commonly used in isolation and identification of ALV associated with clinical disease in chickens. PMID- 19184849 TI - H7N1 avian influenza in Italy (1999 to 2000) in intensively reared chickens and turkeys. AB - From the end of March to the beginning of December 1999, an epidemic of low pathogenicity avian influenza (LPAI) affected the industrial poultry population of northern Italy. The virus responsible for the epidemic was subtyped as H7N1 with an intravenous pathogenicity index (IVPI) of 0.0, and a deduced amino acid sequence of the region coding for the cleavage site of the haemagglutinin molecule typical of low pathogenicity viruses. The circulation of the virus in a susceptible population for several months caused the emergence of a highly pathogenic virus with an IVPI of 3.0 and the presence of multiple basic amino acids in the deduced amino acid sequence for the cleavage site of the haemagglutinin molecule. Over 13 million birds were affected by the epidemic and, in the present paper, we report the results of the clinical, virological and histopathological investigations performed on affected chickens and turkeys. Clinical, gross and microscopic lesions caused by LPAI were more severe in turkeys than in chickens, while highly pathogenicity avian influenza (HPAI) caused similar mortality rates in both species. Current European legislation considers LPAI and HPAI as two completely distinct diseases, not requiring any compulsory eradication policy for LPAI but enforcing eradication for HPAI. In the Italian 1999 to 2000 epidemic, LPAI mutated to HPAI in a densely populated area, causing great economic losses. A reconsideration of the current European Union legislation on avian influenza, including LPAI of the H5 and H7 subtypes, could possibly be an aid to avoiding devastating epidemics for the poultry industry. PMID- 19184850 TI - Avian pneumovirus infection of laying hens: experimental studies. AB - Administration of a virulent strain of avian pneumovirus (APV) to specific pathogen free laying hens by the oculonasal route failed to induce a drop in egg production or any adverse effects on eggshell quality. However, intravenous (i.v.) inoculation of the same strain caused a substantial drop in egg production and a high incidence of soft and thin-shelled eggs. Some respiratory signs were also observed and the hens appeared sick, with diarrhoea being observed in approximately one-half of the hens between 4 and 11 days post-inoculation (p.i.). APV antigen was detected in the oviduct epithelium up to 9 days p.i. This challenge model was then used to investigate the efficacy of live attenuated turkey rhinotracheitis (TRT) vaccine administered alone at 1 day old, or an inactivated TRT vaccine (at 16 weeks), or a combined programme using both vaccines, in protecting against this challenge. Neither the live nor the inactivated vaccine alone protected against clinical signs (respiratory infection or diarrhoea). However, the inactivated, but not the live, vaccine did protect against the effect of the i.v. challenge on laying performance. In contrast, the combined vaccination programme protected completely against both clinical signs and poor egg-laying performance. This protection lasted until at least 60 weeks of age. On the basis of the results with this experimental model, it is concluded that the use of live priming followed by administration of inactivated TRT vaccine is necessary to provide complete protection of laying chickens against APV challenge. PMID- 19184851 TI - Diagnosis of skin lesions in condemned or downgraded broiler carcasses - a microscopic and macroscopic study. AB - Eight hundred skin samples from broiler carcasses condemned or downgraded for skin lesions were collected at five processing plants. Histologically, 45.25% were cellulitis, 19.00% were atypical fowlpox, 3.25% were dermal squamous cell carcinoma (DSCC), 17.00% were non-specific dermatitis, 0.5% were focal haemorrhages and 0.25% were haemangiomas. Of the samples, 14.75% were histologically normal. Macroscopically, increased thickening of the skin was a frequent finding occurring in all the skin diseases, as well as skin discoloration, dark nodules in feather follicles and crusts. Skin scratches were associated with all of the diseases, except DSCC. Crater-like ulcers occurred in DSCC and fowlpox. Cellulitis was more frequent on the abdomen, fowlpox on the dorsum, DSCC in thighs, and non-specific dermatitis on the dorsum, thighs, legs and breast. It was concluded that macroscopic changes are not specific and examinations at slaughterhouses do not allow classification of broiler skin diseases. Histological examination appears to be an important tool to provide more accurate data. PMID- 19184852 TI - High susceptibility of partridges ( Perdix perdix ) to toxoplasmosis compared with other gallinaceous birds. AB - Partridges ( Perdix perdix ), chukars ( Alectoris chukar ), wild guineafowl ( Numida meleagris ), wild turkeys ( Meleagris gallopavo ) and chickens ( Gallus domesticus ) were inoculated per os with 103 or 105 Toxoplasma gondii oocysts (K7 strain). Two of five partridges fed 103 oocysts and six of eight partridges fed 105 oocysts died between day 6 and 16 post-inoculation (p.i.); no clinical symptoms were observed in surviving birds. Antibodies to T. gondii were detected in the birds by the indirect fluorescence antibody test (IFAT) first on day 7 p.i. On days 14, 21 and 28 p.i. (end of the experiment), antibodies were found in all partridges, chukars, guineafowl and turkeys. In chickens, IFAT antibodies were first detected on day 14 p.i., and all chickens were serologically positive on days 21 and 28 p.i. Bioassay in mice revealed T. gondii in the brain, liver, spleen, heart and leg muscles of all partridges and chukars. Enteritis was the most striking lesion in partridges that died. Results indicated that partridges are highly susceptible to toxoplasmosis, while chukars, wild guineafowl and turkeys seem to be less susceptible. Chickens are highly resistant to T. gondii infections. PMID- 19184853 TI - The association of various isolates of Escherichia coli from the United States with induced cellulitis and colibacillosis in young broiler chickens. AB - An experiment was conducted to observe the effects of 10 different avian Escherichia coli isolates in 3-day post-hatch broiler chicks after subcutaneous administration. Isolates were originally obtained from various avian sources throughout the US. Chicks were injected subcutaneously on the ventral surface and necropsied at 7-day intervals for 3 weeks. Cellulitis was produced in all treatments receiving E. coli of cellulitis origin, with the highest incidence occurring 2 weeks post-infection in birds that received an isolate recovered in a previous challenge experiment. Cellulitis was also observed at week 1 post infection in a small percentage of the birds in two of the treatments receiving E. coli of enteric origin, although lesions disappeared from the group after week 1 post-infection. Septicaemia was the most frequent sequel to challenge and occurred regardless of which isolate was injected. Chicks exposed to cellulitis origin isolates developed septicaemia more frequently than birds challenged with E. coli of non-cellulitis origin. The data implies that cellulitis is unlikely to occur early in the bird's life, since young birds exposed to E. coli frequently develop septicaemia. PMID- 19184854 TI - Evaluation of the effect of mannan-oligosaccharides on the competitive exclusion of Salmonella Enteritidis colonization in broiler chicks. AB - This study evaluated the protection against colonization by Salmonella enterica serovar Enteritidis in the chick's caecum when dosed with caecal contents from hens fed dietary carbohydrates based on mannose. Protection was investigated by comparing the effects of four suspensions of hen caecal contents (HCC) fed diets supplemented with 2.5% D -mannose (MAN-HCC), 2.5% mannoseoligosaccharide (MOS HCC), 2.5% palm kernel meal (PKM-HCC) or unsupplemented mash (MashHCC). Four trials to quantify and compare their effectiveness, showed that chicks were better protected when given MOS-HCC or PKM-HCC combined with diets supplemented with the same oligosaccharide given to the hens. Protection remained with the caecal contents diluted up to 106-fold but was reduced or lost at higher dilutions, with better protection produced by the addition of carbohydrates to the diet. In conclusion, this study showed that HCC from hens fed diets supplemented with MOS or PKM were more effective against S. Enteritidis colonization in chicks than the HCC from hens fed unsupplemented mash. PMID- 19184855 TI - Pathogenicity of Salmonella enteritidis phage types 4, 8 and 23 in specific pathogen free chicks. AB - The pathogenicity of two isolates of Salmonella enterica serovar Enteritidis (SE) phage type (PT) 4, three of PT8 and one of PT23 was investigated in groups of 1 day-old specific pathogen free White Leghorn chicks. Two groups were crop gavaged with each culture but at two different doses. Two additional groups were given Salmonella enterica serovar Pullorum (SP) at similar doses and one further group served as uninoculated controls. Body weights were recorded at 14, 21, and 28 days postinoculation (d.p.i), and mortality was monitored throughout. In most treatment groups, the average body weights were significantly lower than the controls. Birds inoculated with SP had the highest mortality followed by those given SE PT4 of human or chicken origin. At 14 and 21 d.p.i., four chicks from each group were killed and examined for gross lesions. Selected tissues were collected for histopathology and cultured for bacteria. Dead birds had fibrinous exudate in the pericardium and also, in a few, on the liver capsule. They had enlarged livers, sometimes with congestion and white foci. At 7 d.p.i., several birds, especially those inoculated with SE PT4, had retained yolk sacs containing coagulated material. Microscopic lesions of pericarditis, myocarditis, hepatitis, splenitis, peritonitis and enteritis were present at 7 d.p.i. in most birds inoculated with SE, but was greatly variable at 14 d.p.i.. This study shows that 1-day-old SPF chicks are susceptible to various phage types of SE, with yolk-sac infection as the most prominent feature. PMID- 19184856 TI - Polymyxin E-1 (colistin sulphate) (neuro-)intoxication in young ostriches ( Struthio camelus spp.). AB - Colistin (polymyxin E) is a cyclic polypeptide with a potent bactericidal action against most gramnegative bacilli. When used parenterally, polymyxins should be given with great care as they have a very small safety range, and easily induce neurotoxicity and nephrotoxicity. A dose of 39.5mg/kg body weight colistin sulphate injected subcutaneously induced rapid (within 1 to 3 h) mortality in young ostriches. Clinical signs of apathy, lethargy and hypotonia indicative of neurotoxicity of the compound were observed. At postmortem, vascular congestion of brain vessels was seen while, on histology, severe acute oedema was present in the epicardium and the intestinal serosa. Congestion of villi, swelling and vacuolization of the plexus ofAuerbach, as well as intermuscular and perivascular oedema of the heart, were also observed. In view of our observations in ostriches and in other species studied, a dose of >5mg/kg body weight polymyxin E is not considered safe for parenteral administration in ostriches. PMID- 19184857 TI - Sequence analysis of the cleavage site-encoding region of the fusion protein gene of Newcastle disease viruses from India and Nepal. AB - Five field isolates of Newcastle disease virus, including one from a pigeon from the Indian subcontinent, along with three vaccine strains have been characterized by sequence analysis of the fusion protein (F) gene in the region encoding the F 2 -F 1 cleavage site. Based on the amino acid sequence present at the cleavage site and on the percent divergence at nucleotide and amino acid levels, three field isolates could be classified as velogenic and two were of lentogenic pathotypes. The velogenic pathotypes had the sequence RRQK/RRF at the cleavage site, while the lentogenic strains had GRQA/GRL at the corresponding position. PMID- 19184858 TI - Target cells for duck enteritis virus in lymphoid organs. AB - Duck enteritis virus (DEV), a herpesvirus, has been shown to cause lymphoid organ atrophy and immunosuppression in white Pekin ducklings. The cells that support virus replication and could be important for immunosuppression were identified in this study. Lymphoid organs of white Pekin ducks infected at 2 weeks of age were collected at 3, 6, 8 and 10 days post-inoculation (d.p.i.). Frozen sections were double-stained for DEV-infected (DEV+) and epithelial cells, DEV+ and CD3+ cells or DEV+ and B cells. DEV antigen was detected in the spleen, thymus and bursa for 3, 6 and 8 d.p.i., respectively. DEV antigen was demonstrated in epithelial cells of all examined lymphoid organs. B cells were irreversibly depleted from bursa; however, the depletion in the spleen was only for 8 d.p.i. Depletion of CD3+ cells was only observed in the thymus. These data show that the target cells for DEV are epithelial and B cells. PMID- 19184859 TI - Relationship between egg size and subgroup J avian leukosis virus in eggs from broiler breeders. AB - Hatching eggs from three broiler breeder flocks that had experienced losses from myeloid leukosis were tested for infection with avian leukosis virus of subgroup J (ALV-J). Sufficient eggs were positive in two flocks to relate infection to egg weight. Allantoic fluid, embryonic tissue and yolk were collected after 18 days of incubation. The albumen and allantoic fluid were tested by enzyme-linked immunosorbent assay (ELISA) for group-specific (gs) antigen and all specimens were inoculated onto cell cultures to test for virus by immunofluorescence assay. Virus detected was identified as ALV-J by polymerase chain reaction techniques. The percentage of eggs that tested positive for gs antigen and virus was higher in those that weighed under 60 g than in heavier eggs (P < 0.01). In one flock, antibody to ALV-J was detected by ELISA in yolk from 14 and 43% of the eggs that tested positive or negative for virus, respectively. Testing the same eggs for antigen, virus and antibody should be useful for establishing the status of infection of the hens. PMID- 19184860 TI - Effect of Cryptosporidium baileyi in specific pathogen free chickens vaccinated (CVI988/Rispens) and challenged with HPRS-16 strain of Marek's. AB - This study was performed to examine the effect of Marek's disease virus (MDV) serotype 1 vaccine (CVI988/Rispens) on the pathogenicity of Cryptosporidium baileyi , and to determine whether C. baileyi infection could prevent the development of vaccinal Marek's disease (MD) immunity in specific pathogen free (SPF) chickens. Sixty-eight SPF homozygous B13 White Leghorn chickens were divided into seven groups. C. baileyi was orally administered at 5 days of age (day 4) in chickens infected with Rispens vaccine at day 0 or at day 8 and challenged with HPRS-16 strain of oncogenic MDV at day 15. Relevant control groups were constituted. The chickens were kept in isolators until the end of the experiment at day 62. The parameters evaluated were clinical signs, kinetics of oocyst shedding, mortality, macroscopic and microscopic lesions, cryptosporidia location in various organs and serum anti- C. baileyi antibodies at days 42 and 62. Our results show that C. baileyi , which is considered to be non-pathogenic when inoculated orally, may become highly pathogenic. It induced severe mortality and developed in organs other than classical target sites when chickens were vaccinated with Rispens vaccine and challenged with the HPRS-16 strain of MDV.However,parasite infection does not prevent the induction of vaccinal immunity for MD. Our results also show that vaccination of B13 chickens at hatching induces higher protection against challenge with HPRS-16 MDV at day 15 than vaccination at day 8. PMID- 19184861 TI - Isolation of a highly pathogenic influenza A virus of subtype H7N3 from a peregrine falcon ( Falco peregrinus ). AB - A peregrine falcon ( Falco peregrinus ) was presented to the Al Safa Falcon Clinic in Dubai, UnitedArab Emirates unable to stand. Four hours after hospitalization, the bird died despite supportive care and calcium disodiumedetate treatment. The falcon had been on a hunting trip to Syria with its owner of 2 years, prior to its death. The carcass was submitted to the Central Veterinary Research Laboratory in Dubai where it was subjected to postmortem examination. Investigations resulted in the isolation of an influenza A virus subtype H7N3, which proved to be highly pathogenic for chickens. PMID- 19184862 TI - Concurrent occurrence of salmonellosis, colibaccillosis and histomoniasis in a broiler flock fed with antibiotic-free commercial feed. AB - A flock of 3-week-old broiler chickens fed with antibiotic-free commercial feed developed cyanotic combs, depression, extended abdomens, reddened abdominal skin, faeces-stained vents, and diarrhoea. By the end of week 6, mortality reached 19.3%, and important lesions seen were ascites, airsacculitis, swollen/congested kidneys, fibrinous perihepatitis, fibrinous pericarditis with or without hydropericardium, haemorrhagic enteritis-typhilitis and ballooned caeca. Salmonella typhmurium var Copenhagen and Escherichia coli were isolated from livers, hearts, intestines and caeca. Histomonas spp. were detected only in caeca. Thus, concurrent occurrence of salmonellosis, colibacillosis and histomoniasis was diagnosed. PMID- 19184863 TI - Highly pathogenic avian influenza (H7N1) in ostriches (Struthio camelus). AB - The clinical, virological and pathological findings observed in a natural outbreak of highly pathogenic avian influenza in intensively farmed ostriches (Struthio camelus) are reported. Clinical signs characterized by anorexia, depression, nervous and enteric signs were observed in young birds, which resulted in death of 30% of the affected birds. Virus isolation performed in accordance with the guidelines listed in European Union Directive 92/40/EEC yielded an influenza A virus of the H7N1 subtype with a deduced cleavage site motif containing multiple basic amino acids, typical of highly pathogenic viruses. Gross lesions, mainly haemorrhagic enteritis and liver degeneration and necrosis, were confirmed by histopathology and immunohistochemistry, resulting in the detection of necrotic lesions and influenza A nucleoprotein in selected organs. The findings reported indicate that ostriches are susceptible to highly pathogenic avian influenza. PMID- 19184865 TI - International meetings. PMID- 19184866 TI - WVPA Matters. PMID- 19184867 TI - Newcastle disease virus: an evolving pathogen? AB - Australia experienced outbreaks of virulent Newcastle disease (ND) in chickens in the state of New South Wales in the years 1998, 1999 and 2000. The disease had occurred previously in Australia in 1930 and 1932 but the country was free of it until the recent outbreaks. Avirulent strains of Newcastle disease virus (NDV) were detected in 1966 and, during the next two to three decades, strains (so called lentogenic strains) able to induce mild respiratory disease equivalent to that induced by vaccine strains such as LaSota were also detected. Nucleotide sequence analysis of the genes encoding the haemagglutinin and fusion proteins of Australian isolates of the virus during this time demonstrated that Australian chicken strains of NDV could be differentiated from NDV isolated elsewhere. Analysis in this way demonstrated that NDV isolates causing the recent outbreaks of virulent disease were Australian viruses that were so closely related to a recognized Australian lentogenic strain, termed the Peat's Ridge strain, that it was considered to be the precursor of the virulent virus. The outbreaks of virulent disease in 1998 and 1999 were controlled by an official "stamping out" eradication campaign. This was subsequently replaced by strategic use of ND vaccines when virulent virus was again detected on some farms that had been restocked following depopulation. The national situation with regard to ND is now being assessed through a structured national survey of ND viruses, particularly to determine the distribution of the precursor strain. No new outbreaks of virulent ND have been recognized since February 2000, although immunization of flocks in areas where the disease was recognized has occurred. PMID- 19184868 TI - An in vitro and in vivo evaluation of the virulence of egg drop syndrome virus for the chicken reproductive tract. AB - The virulence of strains of egg drop syndrome (EDS) 1976 virus for the female reproductive tract of chickens was assessed in vitro using oviduct organ cultures (OOC) prepared from precociously induced oviducts in young chicks by oestrogen treatment. Ciliostasis, haemagglutination and virus titres in infected OOC supernatants, histology and immunoperoxidase test results indicated the pathogenic ability of the four viruses for the precocious oviducts. One of the isolates, EDS TN4, produced higher virus titres in the supernatants of infected OOC and more severe glandular atrophy and necrosis, but caused slightly delayed ciliostasis. When this isolate was used in vivo, virus could not be detected by haemagglutination, but was detected in a few birds using a polymerase chain reaction on the allantoic fluids of infected duck embryos. Ciliostasis of OOC and histological lesions were confined to early stages of infection. This technique could be a pointer to possible variations in virulence of EDS virus isolates, and warrants further investigation. The potential value of OOC from young chickens for EDS diagnosis is emphasized. PMID- 19184869 TI - Genomic characterization of Indian isolates of egg drop syndrome 1976 virus. AB - Five Indian isolates of egg drop syndrome (EDS) 1976 virus and the reference strain 127 were compared by restriction enzyme analysis of viral DNA, and the hexon gene amplified by polymerase chain reaction. Using these techniques, no differences were seen among these viruses. However, partial sequencing of the hexon gene revealed major differences (4.6%) in one of the isolates sequenced, EDS Kerala. Phylogenetic analysis also placed this isolate in a different lineage compared with the other isolates. The need for constant monitoring of the genetic nature of the field isolates of EDS viruses is emphasized. PMID- 19184870 TI - Virulence of Pasteurella multocida subsp. multocida isolated from outbreaks of fowl cholera in wild birds for domestic poultry and game birds. AB - Chickens, turkeys, partridges and pheasants were experimentally infected with Pasteurella multocida subsp. multocida to investigate whether outbreaks of fowl cholera in avifauna might represent a risk for organic, backyard and industrial poultry production. Birds were infected intra-tracheally with a strain of P. multocida subsp. multocida (40605-1) isolated from outbreaks of fowl cholera in wild birds in Denmark. P. multocida subsp. multocida strain P-1059 was included as a reference strain. The outbreak strain was highly virulent for turkeys, partridges and pheasants, while chickens were more resistant. The present findings underline the importance of wild birds as a reservoir for P. multocida. Intratracheal challenge proved useful for studying the virulence of P. multocida. PMID- 19184871 TI - Amyloid A amyloidosis in non-infected and avian leukosis virus-C persistently infected inbred ducks. AB - The breeding history of the first inbred strain of Khaki Campbell ducks is presented. The genetic homogeneity of this strain was tested on the basis of serum amyloid A (SAA) polymorphism and it was established that it harbours only the SAA allele A, which is expressed in liver, lung and bursa of Fabricius tissues. Pathogenic changes in control and avian leukosis virus-C (ALV-C) persistently infected ducks were evaluated during the period spanning 1 to 10 months after hatching. In both groups, AA amyloidosis was revealed and characterized. In spite of the inbred nature of animals, the incidence of amyloid A deposition varied among experiments, suggesting that additional non-genetic factors are involved. Similar variation was found in ALV-C persistently infected ducks, where only in one out of three experiments was the incidence ofAA amyloidosis significantly higher than in controls. PMID- 19184872 TI - Reovirus infection in two species of Psittaciformes recently imported into Italy. AB - An outbreak of reovirus infection with high mortality in two groups of recently imported psittacine birds is reported. The disease in the two species involved, African grey parrots (Psittacus erithacus erithacus) and Australian king parrots (Alisterus scapularis), had differences in clinical presentation and gross lesions. Reovirus particles were observed by electron microscopy and ultrastructural examination of tissues, and two viruses were isolated in cell culture, one from each bird species. Both isolates were studied by cross neutralization with antisera against reference avian reoviruses isolated from chickens and parrots, and were found to have the greatest similarity to viruses isolated from a budgerigar and a southern screamer. PMID- 19184873 TI - Lockjaw syndrome in cockatiels associated with sinusitis. AB - Two cases where multiple juvenile cockatiels exhibited inappetance, depression, upper respiratory signs, and "lockjaw" are described. Symptoms progressed over several weeks until all birds died, in spite of antibacterial therapy. Seven affected birds from each case were submitted for diagnostic evaluation. Microscopically, all birds had necrotizing rhinitis and sinusitis, as well as myositis, perineuritis and osteomyelitis affecting the jaw muscles and cranial bones. Multiple bacterial agents were isolated from the lungs and sinuses in both cases. Juvenile cockatiels appear to be particularly susceptible to temporomandibulitis, temporomandibular joint rigidity, or "lockjaw". Once chronic inflammation and fibrosis develop, it appears unlikely that jaw mobility can be restored. PMID- 19184874 TI - Rapid identification of "very virulent" strains of infectious bursal disease virus by reverse transcription-polymerase chain reaction combined with restriction enzyme analysis. AB - A reverse transcription-polymerase chain reaction (RT-PCR) combined with restriction enzyme analysis (REA) was developed for differentiation of classical virulent (cv) and very virulent (vv) strains of infectious bursal disease virus (IBDV). The VP2 gene was used for primer annealing to amplify the hypervariable region. An amplification product was obtained with serotype 1 and serotype 2 strains of IBDV.The restriction enzymes SacI and BspMI were used to identify and to differentiate the serotype 1 strains: SacI only cleaved cvIBDV RT-PCR products, whereas products obtained with vvIBDV strains were only cleaved with BspMI; serotype 2 strain products were not cleaved by either of these restriction enzymes. RT-PCR combined with REA as described in this study is thus suitable to detect IBDV of both serotypes and to distinguish rapidly between cvIBDV,vvIBDV and serotype 2 IBDV.Our investigation of a total of 11 different IBDV isolates and available nucleotide sequence data of other isolates indicate that the application of this protocol might be a useful tool for the rapid identification of vvIBDV isolates, a prerequisite for the effective control of this economically important virus infection of commercial poultry. PMID- 19184875 TI - Extrahepatic bile duct malformation causing intrahepatic cholangiocellular proliferation with fibrosis in broiler chickens. AB - Malformation of the extrahepatic biliary tracts was observed in six of 88 enlarged, firm and discoloured livers collected at a slaughterhouse. Histologically, the six livers were characterized by diffuse cholangiocellular proliferation with fibrosis, extremely similar to the condition induced by bile duct ligation. These findings suggested that some cases of cholangiohepatitis in chickens are caused by biliary malformations with bile stasis. PMID- 19184876 TI - Detection of infectious bronchitis virus in experimentally infected chickens by an antigen-competitive ELISA. AB - An antigen-competitive enzyme-linked immunosorbent assay (Ag-C-ELISA) was developed for the detection of infectious bronchitis virus (IBV) antigens, M41 strain, in tissues from experimentally infected chickens, or in allantoic fluid harvested from inoculated embryonated eggs. The detection limit of IBV in the Ag C-ELISA was 10(4.1) median embryo infective doses (EID(50))/well. Tracheal and lung samples from chickens vaccinated with 10(2.5) EID(50) of live attenuated infectious bronchitis (H120) vaccine were negative in the direct detection Ag-C ELISA. The results indicate that the Ag-C-ELISA has the potential to detect IBV, either directly in tissue samples or when combined with the passage of material in embryonated eggs, thereby constituting an alternative method for the diagnosis of IBV. PMID- 19184877 TI - Severely impaired production performance in broiler flocks with high incidence of Clostridium perfringens-associated hepatitis. AB - Historical meat inspection data and production performance data collected from a large poultry processing plant in Norway were used to study the association between Clostridium perfringens infection and production performance in commercial broiler flocks. The study period was limited to the first 2.5 years after withdrawal of the in-feed antibiotic avoparcin, when necrotic enteritis was frequently seen. High levels of C. perfringens-associated hepatitis at slaughter were used as an indicator of C.perfringens infection. The contribution margins were 25 to 43% lower from flocks with high levels of C. perfringens-associated hepatitis than those with low levels. Impaired feed conversion ratio and reduced bodyweight at slaughter were major causes of the losses, while condemnations per se contributed to one-fifth of the losses in profit. Incidence of liver lesions showed remarkable variation throughout the study period, and there was considerable difference in frequency of liver lesions between farms. PMID- 19184878 TI - Assessment of the pathogenicity of an emu-origin influenza A H5 virus in ostriches (Struthio camelus). AB - Ostriches were inoculated with a laboratory-derived highly pathogenic avian influenza (HPAI) virus of emu origin, A/emu/TX/39924/93 (H5N2) clone c1B. The aim of this study was to evaluate the pathogenicity of this isolate for ostriches and assess the ability of routine virological and serological tests to detect infection. Avian influenza virus (AIV) was isolated from cloacal and tracheal swabs from 2 to 12 days post-infection. AIV was also isolated from brain, thymus, eyelid, spleen, ovary/testis, liver, air sac, proventriculum, duodenum, caecal tonsil, heart, pancreas, kidney, nasal gland and lung. Virus isolation was also possible from swabs of the luminal surfaces of the cloaca, jejunum, lower ileum, bursa of Fabricius, trachea and bone marrow. Birds seroconverted as early as 7 days post-infection. This study suggests that HPAI virus of emu origin replicates extensively in infected ostriches without causing significant clinical disease or mortality. PMID- 19184879 TI - Avian pox infection in Spanish Imperial eagles (Aquila adalberti). AB - A cutaneous lesion, previously known as "warts", affecting the featherless parts of face and legs has long been recognized in juvenile Spanish Imperial eagles (Aquila adalberti). This paper describes the presentation, microbiological, histopathological, and electron microscopic findings of lesions and diagnosis of poxvirus infection in nine juveniles. Lesions consisted of single or multiple nodules with a crust and surrounded by skin swelling. Seventy-eight percent of the swabs taken from lesions yielded bacterial growth, with Escherichia coli being the most common bacterium isolated. Histopathology revealed typical pox lesions in all cases. Histopathological changes found consisted of proliferative epithelium, with ballooning degeneration of keratinocytes and lymphocyte infiltrates extending into underlying dermis. Avianpox virus was confirmed by the presence of eosinophilic intracytoplasmatic inclusion bodies in the affected cells on light microscopy, and diagnosis confirmation was performed by electron microscopy of biopsies from all nine eagles. PMID- 19184881 TI - International meetings. PMID- 19184883 TI - Addendum. PMID- 19184884 TI - A nomenclature for avian coronavirus isolates and the question of species status. AB - Currently, there is no agreed naming system for isolates of infectious bronchitis virus (IBV), whose host is the domestic fowl (Gallus gallus domesticus). A uniform, informative system for naming IBV isolates would be very helpful. Furthermore, the desirability of a single naming system has become more important with the recent discoveries that coronaviruses with genome organizations and gene sequences very similar to those of IBV have been isolated from turkeys (Meleagris gallopavo) and pheasants (Phasianus colchicus). To date, no genetic features have been found that are unique to turkey isolates and to pheasant isolates that would permit unequivocal differentiation from IBVs. Should the avian coronaviruses from turkeys, pheasants and other birds each be considered as distinct coronavirus species? Or should avian coronaviruses that have gene sequences similar to those of IBV be treated as host-range variants of IBV or, more objectively, as host range variants of a species that might be called avian coronavirus (ACoV)? Clearly, the topic of avian coronavirus species differentiation requires debate. For the moment, a naming system for avian coronavirus isolates is overdue. Increasingly, papers will include data of coronaviruses isolated from more than one species of bird. It is desirable to have a nomenclature for avian coronaviruses that indicates the host species of origin. Furthermore, it would be helpful if the name of an isolate included the country/region of origin, an isolate number and the year of isolation. The names of avian paramyxovirus (APMV) and avian influenza virus (AIV) isolates have long since contained this information; I suggest that we adopt a similar convention for isolates of avian coronaviruses. For example, the D274 isolate of IBV could be named chicken/Netherlands/D274/78. Representatives of avian coronaviruses from turkey and pheasant would include turkey/United States(Nc)/NC95/95 and pheasant/UK/750/83. Two upper case letters would be used to denote country of isolation, whereas one upper and one lower case letter would be used to indicate state or province, e.g. Nc, North Carolina. The full-length names could be abbreviated, when desired, similar to the convention used for AIV isolates, e.g. chNL78, tyUS(Nc)95 and phUK83. If the serotype of an isolate has been clearly established, this might be included in the name at end, like the serotype designation of AIVs, e.g. chicken/China/NRZ/91 (Mass.) for the Chinese isolate of the Massachusetts serotype. This suggested naming system for isolates is essentially neutral with regard to whether viruses from different bird species should be considered as different coronavirus species or simply as variants of just one avian coronavirus species. In my opinion an informative nomenclature for avian coronavirus isolates is required now, to improve communication, and need not be delayed until a decision on the definition of coronavirus species has been made. PMID- 19184885 TI - Detection and differentiation of Newcastle disease virus (avian paramyxovirus type 1). AB - Substantial variation in the virulence of Newcastle disease virus (NDV) isolates means that the detection of NDV or evidence of infection is insufficient for an adequate diagnosis, as control measures for avirulent viruses are very different to those for virulent viruses. Diagnosis therefore requires further characterization, at least as to whether an isolate is virulent or avirulent. Conventional detection and differentiation of ND viruses is perceived as slow, laborious and requiring an undesirable use of in vivo techniques. In addition, further characterization is needed to give greater information on origin and spread. This review concentrates on the application of monoclonal antibody and molecular biological approaches. Panels of monoclonal antibodies were a major advance for the characterization of NDV isolates, although confirmation of virulence for poultry still required in vivo testing. As molecular-based techniques become easier and more reliable, they are likely to supersede the use of monoclonal antibodies, especially for characterizing viruses for epidemiological purposes. The attraction of molecular-based techniques is that they may be able to cover all three aspects of Newcastle disease diagnosis (detection of virus, characterization, including inference of virulence, and epidemiology) quickly, accurately and definitively in a single test. A number of approaches based on the reverse transcriptase polymerase chain reaction have been developed, with subsequent analysis of the product by restriction enzyme analysis, probe hybridization and nucleotide sequencing. Although extensive variation among NDVs still poses technical problems, the real and potential advantages of a molecular biological approach to Newcastle disease diagnosis appear to be overwhelming. PMID- 19184886 TI - Experimental Salmonella enterica serovar Pullorum infection in two commercial varieties of laying hens. AB - An experiment was carried out to investigate the biology of Salmonella Pullorum in two varieties of laying hens, from 5 days of age up to 9 months. One variety was resistant to systemic salmonellosis (light layers producing white eggs) and the other was considered susceptible (brown layers producing brown eggs). The brown birds were more affected by the infection, showing signs of clinical disease in the first month of life. Later, these signs disappeared, but postmortem examination revealed persistent gross pathological changes in the liver, spleen, heart and ovary. The rapid agglutination test detected reactors throughout the experiment, with the strongest agglutination from 1 to 7 months post-infection. S . Pullorum was isolated from some of the organs and the eggs laid throughout the experiment. The relationship between white birds and S . Pullorum was less intense, and there were no noticeable signs of disease. There were few gross pathological changes, and the bacteria were isolated infrequently and only for a brief period after infection, although contaminated eggs were laid by these birds. The strongest serological response in the white chickens occurred between the second and the fifth month post-infection. PMID- 19184887 TI - The pathology of normobaric oxygen toxicity in budgerigars (Melopsittacus undulatus). AB - The effects of normobaric oxygen exposure were investigated in budgerigars (Melopsittacus undulatus). Sixty birds were randomly divided into four equal groups of 15. These groups were randomly allocated as control, acute exposure, repeated acute exposure or chronic exposure. Control birds were exposed to 72 continuous hours of 21% oxygen in a sealed, enclosed chamber. Acute exposure, repeated acute exposure and chronic exposure groups were exposed oxygen at minimum concentration of 95% for a single 3-h period, a 3-h period daily for three sequential days, or a single 72-h period in a sealed, enclosed chamber, respectively. Oxygen exposure resulted in significant alteration in the histological morphology of respiratory exchange tissue, with severe oedema, and inflammatory cell infiltration. Electron micrographs revealed thickening of the blood-gas barrier with the tissue harmonic thickness increasing from 226 +/-90 nm in control birds to 639 +/-393nm following repeated acute exposure, with the total harmonic thickness increasing from a control value of 345 +/-146nm to 837 +/-423 nm at the same time. Chronic oxygen exposure resulted in significant changes in cell morphology including thickening of endothelial cells, ruffling of type I respiratory endothelial cells and interstitial vacuolation. These results indicate that budgerigars undergo significant morphological and ultrastructural changes in respiratory exchange tissue following exposure to 100% oxygen. PMID- 19184888 TI - Protection of chickens from infectious laryngotracheitis with a recombinant fowlpox virus expressing glycoprotein B of infectious laryngotracheitis virus. AB - Infectious laryngotracheitis (ILT) is an economically important disease of chickens caused by a type I gallid herpesvirus, infectious laryngotracheitis virus (ILTV). The vaccines currently available are modified live viruses, which are effective in preventing disease outbreaks. However, they have often been associated with a variety of adverse effects including spread of vaccine virus to non-vaccinates, inadequate attenuation, production of latently infected carriers, and increased virulence as a result of in vivo passage. In this study, a recombinant fowlpox virus expressing glycoprotein B (gB) of ILTV (rFPV-ILTVgB) was constructed. Protection of specific pathogen free (SPF) and commercial chickens from ILT with the rFPV-ILTVgB and commercial ILTV vaccine (Nobilis ILT) were compared after challenge with a lethal dose of virulent ILTV.Both the rFPV ILTVgB- and the Nobilis ILT-vaccinated SPF chickens were completely protected from death, while 90% of the unvaccinated chickens died after challenge. The immunized commercial chickens were also 100% protected with rFPV-ILTVgB, compared with 85% protected with Nobilis ILT. The protective efficacy was also measured by the antibody response to ILTV gB, isolation of challenge virus and polymerase chain reaction amplification of the ILTV thymidine kinase gene after challenge. The results showed that rFPV-ILTVgB could be a potential safe vaccine to replace current modified live vaccines for preventing ILT. PMID- 19184889 TI - Pigeon circovirus infection: pathological observations and suggested pathogenesis. AB - Pigeon circovirus infection (PiCV) was diagnosed by light and transmission electron microscopy in 15 birds from five lofts in western France. Histopathological findings were suggestive of primary bursotropism of pigeon circovirus, followed by secondary systemic spread from the bursa of Fabricius, particularly to non-bursal lymphoid organs. The last stage of the disease was associated with various secondary (particularly bacterial) infections. In situ detection of apoptosis in the bursa of Fabricius indicated that PiCV was concomitant with an increase in bursal lymphocytic apoptotic events related to viral infection and leading to severe acquired immunosuppression. PMID- 19184890 TI - Atypical poxvirus lesions in two Galapagos doves (Nesopelia g. galapagoensis). AB - Two Galapagos doves were received for necropsy. The only lesions detected were small firm foci throughout the lungs, suspected to be neoplastic. Sections of lung tissues showed the foci as dense, solid masses of epithelioid cells. Electron microscopy revealed pox virions in the lesions. This is the first report on Galapagos doves with poxvirus infection in the lungs with tumour-like growth. PMID- 19184891 TI - A longitudinal study of velogenic Newcastle disease virus genotypes isolated in Italy between 1960 and 2000. AB - Thirty-six representative velogenic strains of Newcastle disease virus isolated in Italy since 1960 were characterized by restriction site and partial sequence analyses of the fusion protein gene. Viruses belonging to the six known genotypes of Lomniczi et al . were found. Genotype IV, which was most probably the main epizootic group in Europe before the war, was responsible for outbreaks in the 1960s and persisted until the late 1980s in Italy. An epizootic peak in 1972 to 1974 coincided with the appearance of genotype V viruses that were present for more than a decade. Outbreaks in 1992 were caused by genotype VIIa viruses and were part of a contemporaneous epizootic of Far East origin that affected Western European countries. The Newcastle disease epizootic that commenced in Italy in May 2000 was due to a genotype VIIb virus that is indistinguishable from those causing sporadic outbreaks in Great Britain and Northern Europe in the late 1990s. Isolated cases yielded a variant of genotype VI (reference epizootic: Middle East in the late 1960s) and a group VIII virus (enzootic in South Africa). PMID- 19184892 TI - Relationship between myocardial beta-adrenergic receptor characteristics and the incidence of ascites in broiler chickens. AB - The present study was carried out to determine, first, the cardiac beta adrenergic receptor characteristics of normal chickens and chickens with heart failure and, second, the characteristics of myocardial beta-adrenergic receptors in non-hypertrophied hearts in broilers exposed to two different altitude and temperature programmes. The density of beta-adrenergic receptors was significantly lower in myocardial cells in right ventricular failure birds compared with healthy birds. The binding capacity of beta-adrenergic receptors was reduced in the non-hypertrophied ventricles after exposure to high altitude in the younger but not in older chickens. At both altitudes, beta-adrenergic receptor binding capacity was not significantly different in low temperature compared with normal temperature birds. These data support the hypothesis of beta adrenergic receptor downregulation as one of the mechanisms of cardiac cell adaptation to hypertrophy or hypoxia. Second, the acclimatization to high altitude in older chickens resulted in a normalization of beta-receptor capacity in those broilers that were not affected by right ventricular failure and ascites. PMID- 19184893 TI - The effects of early exposure of cellulitis-associated Escherichia coli in 1-day old broiler chickens. AB - Two experiments were performed to test the effect of various field strains of Escherichia coli of cellulitis origin. In the first experiment, 1-day-old broiler chicks were challenged with one of two E. coli field strains using inoculation routes including oral gavage, swabbing of the navel and subcutaneous injection. No cellulitis lesions were produced, although the birds experienced high levels of septicemia/toxemia, characteristic of colibacillosis. The birds that received the E. coli by subcutaneous injection experienced the highest rate of mortality, while those that were challenged by gavage and those that had their navels swabbed experienced lesser rates of mortality. Birds in the second experiment were challenged at 1 day of age with one of three field strains of cellulitis origin E. coli administered alone or in combination (1:1), which were serially diluted prior to subcutaneous injection. No significant differences in body weight, mortality or cellulitis rates were associated with specific isolates given; however, significant differences were seen with mortality and cellulitis rates according to the dilution of bacteria given. A linear effect was also noted with body weight at 3 weeks, again correlating to the dilution of bacteria that the chicks received. PMID- 19184894 TI - Mortality in Muscovy ducks (Cairina moschata) and domestic geese (Anser anser var. domestica) associated with natural infection with a highly pathogenic avian influenza virus of H7N1 subtype. AB - Among the 413 outbreaks of highly pathogenic avian influenza (HPAI) caused by a virus of the H7N1 subtype, which occurred in Italy during 1999 and 2000, an outbreak diagnosed in a backyard flock was characterized by mortality and nervous signs in ducks and geese. Dead geese (Anser anser var. domestica) and Muscovy ducks (Cairina moschata) were submitted to the laboratory for bacteriological, virological, histological and immunohistochemical investigations. Routine bacteriological tests resulted negative, while a HPAI virus of the H7N1 subtype was isolated from the geese. Pancreatic damage was observed in both the geese and the ducks, and the pancreas was also positive by immunohistochemistry for avian influenza in the geese. Histopathological lesions were observed in the central nervous system of both species, and this result was supported by positive immunohistochemical findings for the presence of the virus. PMID- 19184896 TI - International meetings. PMID- 19184897 TI - Wvpa matters. PMID- 19184899 TI - Erratum. PMID- 19184900 TI - Antibiotic sensitivity and resistance in Ornithobacterium rhinotracheale strains from Belgian broiler chickens. AB - Establishing the antibiotic sensitivity of the avian respiratory pathogen Ornithobacterium rhinotracheale is difficult because of the organism's complex growth requirements and the unusually frequent occurrence of resistance. The minimal inhibitory concentrations of 10 antibiotics were determined for 45 strains of O. rhinotracheale from Belgian broiler chickens collected from 45 farms between 1995 and 1998. They were compared with the type strain, which was isolated from a turkey, and a strain isolated from a rook. All the broiler strains were resistant to lincomycin and to the beta-lactams ampicillin and ceftiofur. Less than 10% of the strains were sensitive to the macrolides tylosin and spiramycin, tilmicosin and flumequine. A few strains were sensitive to enrofloxacin and doxycycline. All strains were sensitive to tiamulin. PMID- 19184901 TI - The role of egg drop syndrome virus in acute respiratory disease of goslings. AB - An outbreak of severe acute respiratory disease characterized by tracheitis and bronchitis was observed in young goslings on a large-scale goose farm in Hungary. Histological examination revealed amphophilic intranuclear inclusion bodies in the superficial epithelial cells of the trachea and bronchi. Adenovirus-like particles were detected by electron microscopy, and the virus isolated from the trachea and the lungs was identified as egg drop syndrome (EDS) virus by serological and genomic examination. The clinical and pathological signs were reproduced by intratracheal administration of the virus isolate to 1-day-old goslings free of EDS antibodies. The presence of EDS virus DNA in different organs of the naturally and experimentally infected goslings was detected by polymerase chain reaction. This is the first report on the involvement of EDS virus in severe respiratory disease of geese. PMID- 19184902 TI - Trematodes of free range reared guinea fowls (Numida meleagris Linnaeus, 1758) in the state of Rio de Janeiro, Brazil: morphology and pathology. AB - An investigation on the digenetic trematodes parasitizing free range reared guinea fowls was conducted in the state of Rio de Janeiro, Brazil, based on clinical examination, necropsies and histopathological findings in 36 guinea fowls. Parasites, prevalence and intensity of infection were, respectively: Paratanaisia bragai, 61.1% and 31 (in each kidney); Athesmia heterolecithodes, 8.3% and 38; Conspicuum conspicuum, 2.8% and 1. A. heterolecithodes was highly pathogenic to guinea fowls, causing severe hepatic lesions. However, C. conspicuum and P. bragai were less pathogenic in the considered intensities of infection. This is the first time that these parasite species have been reported in Brazilian guinea fowls. PMID- 19184903 TI - The origins and biological significance of the coccidial lesions that occur in chickens vaccinated with a live attenuated anticoccidial vaccine. AB - The occurrence of vaccine-induced coccidial lesions in chicks vaccinated with a live attenuated anticoccidial vaccine is demonstrated. Healthy broiler chicks reared on litter to facilitate autoreinfection, but isolated from extraneous coccidial infections, were vaccinated when 8 days old with Paracox. Coccidial lesions were found in chicks between 5 and 23 days after vaccination; 24% of the 87 chicks sampled during 29 days had intestinal lesions of Eimeria acervulina, Eimeria maxima, Eimeria tenella or of undetermined species, mostly (19 out of 21) scored as 1 or 2 on the Johnson & Reid scale of 0 to 4. The other two positive birds had E. tenella lesions, each scored as 3. The lesions observed up to 5 days after vaccination were identified as primary (i.e. a host response to the first vaccinal life cycle) and those observed from 6 days onwards were designated as primary or secondary (i.e. a host response to the second and subsequent vaccinal life cycles) in origin. In the absence of extraneous coccidial infections, none of the lesions observed could have been due to non-vaccinal coccidial infections. No adverse effects on the health or growth rate of the chicks exhibiting primary or secondary lesions in response to vaccination with Paracox were apparent. PMID- 19184904 TI - Observations on the persistence and vertical transmission of Salmonella enterica serovars Pullorum and Gallinarum in chickens: effect of bacterial and host genetic background. AB - Commercial laying hens inoculated with a strain of Salmonella enterica ser. Pullorum when they were 4 days old showed no morbidity, but harboured infection until they came into lay, and then produced S. Pullorum-contaminated eggs and infected progeny. There was limited evidence of transmission of maternal immunity to the progeny. Attempts were made to set up similar infections in hens with Salmonella Gallinarum, but without success. Infection either resulted in clinical disease or elimination of the pathogen. Infection of birds when in lay produced a similar result. The possibility of eggs becoming contaminated with S. Gallinarum after they were laid in the nest box was evaluated but there was no evidence for this. In-bred chicken lines with a SalI-susceptible phenotype showed greater localization of S. Pullorum in the reproductive tract than did a SalI-resistant line. In addition, in-bred birds, which were SalI resistant but showed greater susceptibility to intestinal colonization by Salmonella, infected with S. Gallinarum when they were 1 week old, showed longer term persistence in the liver and spleen than did a resistant line. PMID- 19184905 TI - Infectious bronchitis virus vaccine interferes with the replication of avian pneumovirus vaccine in domestic fowl. AB - Experiments were performed in chickens to ascertain whether application of infectious bronchitis (IB) H120 vaccine had an effect on the replication of an attenuated avian pneumovirus (APV) strain, using as indicators virus detection, humoral antibody responses and clinical protection against in vivo APV challenge. A preliminary experiment demonstrated that pharyngeal swabs were as efficient for recovery of APV as were buccal cavity swabs, and that either site was superior to swabbing the nasal cavity. APV was detected to a similar extent by both a reverse transcriptase-polymerase chain reaction (RT-PCR) and virus isolation; therefore, RT-PCR was used in subsequent experiments. In chickens vaccinated with APV alone, APV was detected by RT-PCR in most birds for 1 week after vaccination. When IB vaccine had been applied 1 week earlier, APV detection was delayed and much reduced. This interference by IBV resulted in a lower APV antibody response to vaccination. Following challenge with virulent APV, birds that had been vaccinated with APV alone were fully protected both clinically and virologically. Chickens that had received both vaccines were still protected clinically, but challenge virus could be detected in some pharyngeal swabs 4 days after challenge. In contrast, the APV vaccine had no effect on either the antibody response to the IB vaccine or the level of protection against IB challenge. It is concluded that IB vaccination interferes with the replication of APV, resulting in a reduction in the antibody response but with no adverse effect on the induction of protective immunity. PMID- 19184906 TI - The use of serotype 1- and serotype 3-specific polymerase chain reaction for the detection of Marek's disease virus in chickens. AB - A serotype 1- and serotype 3-specific detection of Marek's disease virus (MDV) by polymerase chain reaction (PCR) was developed. The sensitivity of the method when applied to cell culture grown virus was comparable with that of cultivation. The method was applied to various tissue samples from chickens experimentally inoculated with serotype 1 or serotype 3 MDV.The serotype 1 strains CVI988 and RB 1B could be detected in feather follicle epithelium up to 56 and 84 days post inoculation (p.i.), respectively, while the MDV-3 serotype was detected until 42 days p.i. The purpose of this study was to develop and evaluate a reliable and easy-to-handle method for surveillance of the occurrence of MDV in chicken flocks. We emphasize the development of a method, which can be applied to types of samples conveniently collected in the field, e.g. feather tips and blood samples. In addition, the PCR was applied to samples collected from four commercial table egg layer flocks of young stock or pullets vaccinated with either serotype 1 (CVI988) or serotype 3 (HVT) vaccine. These flocks had various clinical signs of Marek's disease. MDV-1 was detected in buffy-coat cells, spleen, liver, skin, feather tips and ovaries. The detection of MDV in feather tips appeared to be as sensitive as co-cultivation of buffy-coat cells, although an inhibiting factor was observed in extracts from feather tips of non-white chickens. This inhibition could be overcome in most extracts by applying a bovine serum albumen pretreatment. The PCR proved to be a convenient tool for the monitoring of MDV in the poultry population, and feather tips were the most convenient and sensitive samples. PMID- 19184907 TI - Morphological and molecular characterization of Encephalitozoon hellem in hummingbirds. AB - Microsporidiosis was identified as a cause of enteritis in wild, migratory hummingbirds (Calypte anna). Electron microscopic examinations of parasites showed microsporidian spores with a double spore coat and a polar filament containing four to six coils, compatible with the genus Encephalitozoon. Molecular analysis of ribosomal RNA genes further identified the parasites from droppings and small intestinal segments as Encephalitozoon hellem, genotype I. Microsporidial spores were identified in 19% of droppings from C. anna, Archilochus alexandri and Selasporus sasin using Gram or modified trichrome staining methods. Since E. hellem is an opportunistic pathogen in immunocompromised humans, the pathogenic potential in avian hosts, the zoonotic potential of this parasite, and the role of birds as reservoirs needs to be further explored. PMID- 19184908 TI - Avian pneumovirus infection in broiler chicks inoculated with Escherichia coli at different time intervals. AB - The effects of dual infection of 1-day-old broiler chicks with a chicken isolate of avian pneumovirus (APV) and a pool of pathogenic Escherichia coli strains were studied by supraconjunctival application of the bacteria simultaneously with the virus, or at 4, 7 or 11 days afterwards. When the agents were given together, the clinical disease was significantly more severe than that caused by the virus alone, but when the bacterium was given later the signs were less severe. None of the infections resulted in swollen head syndrome by 32 days. All mixed infections caused moderate to severe congestion in the turbinates, when birds were examined at 32 days of age, at which time no such lesions were present in birds having been infected with APV alone. E. coli was isolated from almost 100% of birds with mixed infections, while rates of those given only E. coli isolation varied between 56 and 67%. Furthermore, E. coli colony counts were consistently higher from mixed infection groups. Virus persistence in the choanal cleft was slightly prolonged in birds with the simultaneous mixed infection. Although the pool of E. coli included O2, O78 and O18 serotypes, only those of the O2 serotype and a small number of untypable strains were re-isolated from selected mixed and single E. coli-infected groups. Mixed APV and E. coli infection did not affect APV enzyme-linked immunosorbent assay antibody titres at 21 or 32 days. Thus, experimental infection of broiler chicks with APV and E. coli, simultaneously or at intervals afterwards, demonstrated a synergistic effect between the two agents, but none of the infection protocols caused swollen head syndrome. PMID- 19184909 TI - Primary target cells of virulent strains of type A influenza virus in chicken embryos. AB - Virulent or avirulent strains of type A influenza virus were inoculated into the allantoic cavities of chicken embryos. The antigens of virulent strains appeared initially in the surface epithelium of the allantoic membrane, then in vascular endothelial cells of the chorioallantoic membrane and visceral organs of the embryos, and then spread to parenchymal cells of many organs. In contrast, the antigens of the avirulent strain were confined to the allantoic membrane. These observations indicate that the primary target of virulent influenza viruses in chicken embryos is vascular endothelial cells, and that the embryos died after systemic viral infection. PMID- 19184911 TI - International meetings. PMID- 19184912 TI - Wvpa matters. PMID- 19184914 TI - Immunization against coccidiosis in pheasants with low-dose live sporulated oocysts of Eimeria colchici. AB - Low-dose immunization of pheasants with live Eimeria colchici oocysts was observed to be effective against massive challenge infection. Fourteen-day-old pheasants, in groups of 28, were orally inoculated with about 100, 250, 500, 1000 and 2000 sporulated oocysts. These birds were then challenged homologously with about 120 000 sporulated oocysts 18 days after the immunization. During the 12 day observation period after the challenge infection, over 92% of the immunized pheasants were still alive, but only 11% of the non-immunized control birds survived the challenge infection. Moreover, the number of oocysts produced after challenge infection of the immunized group was only 10 to 19% of the positive non immunized control group. The crowding effect of E. colchici infection in pheasant was thought to occur based on the significant difference of the relationship between inoculation doses, daily oocysts excretion and the amount of the faeces discharged, as analyzed by Pearson correlation analysis. Our results showed that immunization of pheasants with a low dose of E. colchici oocysts is feasible to help protect the birds from massive homologous infection, which can be fatal. PMID- 19184915 TI - Further studies on vertical transmission and persistence of Salmonella enterica serovar Enteritidis phage type 4 in chickens. AB - One-week-old commercial layers were infected orally with 10(8) colony forming units of Salmonella enterica serovar Enteritidis phage type 4. No mortality was observed. The inoculated organism was isolated in decreasing viable numbers from a number of tissues, particularly the spleen, liver and caeca. Organisms present in the spleen were primarily localized within macrophages. No Salmonella Enteritidis organisms were isolated between 10 and 24 weeks of age, when the experiment was terminated after several weeks of lay. When two groups of adult hens, housed with males, were infected, contaminated eggs were found within 2 weeks of infection in one of the experiments only. Progeny hatched from these eggs showed no mortality unless they were infected artificially with the S. Enteritidis strain. In this case, the percentage mortality fell as the hatches progressed, indicating increasing immunity to infection. The faecal excretion of the inoculated phage type 4 strain by infected but healthy progeny was followed. Although most birds ceased to excrete by 11 to 12 weeks of age, a small number of the birds continued to excrete until they themselves came into lay. The small numbers of birds in which this occurred indicates that tolerance to infection does not occur readily following infection of hens laying fertile eggs or in progeny birds infected before or within hours of hatching. Birds infected when they were less than 24 h old remained persistently infected until they were well into lay. However, control birds infected when 1 week old, on this occasion, showed a high level of excretion until the birds began to lay at 18 weeks. Inbred lines of chickens showing differences in their susceptibility to systemic salmonellosis did not show significant differences in the extent to which S. Enteritidis localized in the organs of the reproductive tract or in the number of infected eggs produced. PMID- 19184916 TI - T-Cell suppression by cyclosporin-A enhances infectious bursal disease virus infection in experimentally infected chickens. AB - In this study, the role of T lymphocytes was investigated in chickens experimentally infected with infectious bursal disease virus (IBDV). Chickens were treated with cyclosporin-A (CS-A), a selective T-cell suppressant drug, by the intramuscular route, starting 3 days before virus infection and every third day thereafter,and infectious bursal disease pathogenesis was compared in such T cell suppressed and intact chickens using a vaccine strain; namely, Georgia and a field isolate of IBDV. Treatment of chickens with CS-A caused a significant suppression of phytohaemagglutinin-A specific proliferative responses of peripheral blood mononuclear cells. The virus neutralizing antibody titres in such CS-A treated chickens were not suppressed. T-Cell suppression resulted in an increase in the severity of gross lesions caused by IBDV and extensive muscular haemorrhages were observed in such chickens between 15 and 21 days post inoculation. Similarly, there was a marked increase in the severity of infectious bursal disease-specific microscopic lesions in the bursa of T-cell suppressed chickens. Consistently higher titres of virus were observed in bursa of CS-A treated chickens. Virus titres were 1 to 2 log(10) higher in the T-cell suppressed chickens as compared with the intact ones. These studies suggest that T cells play a role in limiting the IBDV infection. PMID- 19184918 TI - Parathyroid hormone-related peptide expression in tibial dyschondroplasia. AB - Parathyroid hormone-related peptide (PTHrP) has a key role in the growth of long bones, as it is a negative regulator of growth plate chondrocyte terminal differentiation. We have examined the distribution and gene expression levels of PTHrP in the growth plates of broiler chickens with tibial dyschondroplasia (TD) in order to determine whether increased expression of PTHrP is responsible for the delayed chondrocyte differentiation that is characteristic of this skeletal disorder. PTHrP protein distribution and gene expression levels were assessed by immunocytochemistry and reverse transcriptase-polymerase chain reaction, respectively. In growth plates of normal birds, PTHrP was found to be distributed throughout all maturational zones of the growth plate. In cartilage proximal to the TD lesion, PTHrP immunostaining and the level of PTHrP gene expression were similar to that observed in normal birds. In contrast, many chondrocytes within the centre of the TD lesion stained poorly for PTHrP and this was reflected in the lower levels of PTHrP mRNA detected in lesion cells. These results suggest that alterations in PTHrP distribution and gene expression are not primarily responsible for the delayed chondrocyte differentiation and hypertrophy noted in dyschondroplasia, but are a result of secondary changes due to the pathology of the condition. PMID- 19184917 TI - Nucleotide sequence-based identification of a novel circovirus of canaries. AB - Circovirus-like, spherical particles measuring 16 to 18 nm in diameter were detected in organ homogenates from adult canaries that had died after a short illness characterized by dullness, anorexia, lethargy and feather disorder. A polymerase chain reaction method, based on degenerate primers specific to conserved amino acid sequences in the circovirus replication-associated protein, was used to amplify DNA specific to a novel circovirus, tentatively named canary circovirus (CCV). Sequence analysis of a 510 nucleotide genomic fragment indicated that CCV exhibited 67.4, 64.9, 53.7 and 53.9% nucleotide identities and 70.0, 61.8, 40.4 and 40.1% amino acid identities with columbid (pigeon) circovirus (CoCV), beak and feather disease virus (BFDV), porcine circovirus type 1 and porcine circovirus type 2, respectively. CCV therefore represents a new avian virus of the genus Circovirus of the family Circoviridae, and is more closely related to CoCV than BFDV.The availability of nucleotide sequence data will facilitate the development of DNA-detecting diagnostics with which the prevalence of CCV infections can be assessed. PMID- 19184919 TI - Comparison of virulence and restriction endonuclease cleavage patterns of infectious laryngotracheitis viruses isolated in Korea. AB - Isolates of infectious laryngotracheitis virus (ILTV) obtained from field disease outbreaks in Korea from 1982 to 1998 were compared by virulence testing and by examining restriction endonuclease (REN) cleavage patterns of viral DNA. Based on pathogenicity tests, eight of 11 ILTV strains were classified as virulent, because these strains caused 40 to 80% mortality in specific pathogen free chickens, while three strains were classified as low virulence because these did not cause mortality. The REN cleavage patterns of the low virulence strains were identical with those of two reference vaccine strains, which were of chicken embryo origin. However, the DNA cleavage patterns of the virulent strains differed from those of both the low virulence and the vaccine strains. Furthermore, one virulent Korean strain N87278 had REN cleavage patterns that were clearly different from other virulent strains. In the present study, ILTV strains examined could be classified into two groups (virulent and low virulence strains) by pathogenicity testing, and three groups based on their REN cleavage patterns. These results suggest that most outbreaks of infectious laryngotracheitis were not likely to be associated with vaccine strains, but some were associated with viruses indistinguishable from commercial vaccine strains. At least three genetically distinct groupings of ILTV have been involved in outbreaks of infectious laryngotracheitis in Korea. PMID- 19184920 TI - Efficacy of inactivated infectious bursal disease (IBD) vaccines: comparison of serology with protection of progeny chickens against IBD virus strains of varying virulence. AB - The efficacy of inactivated infectious bursal disease vaccines was determined by measuring both the antibody response of vaccinated chickens and clinical protection of progeny chicks from vaccinated dams. Similar virus neutralizing (VN) antibody titres were obtained in 4-week-old chickens and mature hens after vaccination with one vaccine dose. VN titres below 10 log 2 increased considerably between the fourth and seventh week after vaccination in 4-week-old chickens as well as in mature chickens. All 2-week-old progeny chicks with serum VN antibody titres of at least 9 log 2 were clinically protected against the classical virulent 52/70 infectious bursal disease virus (IBDV) strain, as well as against the very virulent IBDV (vvIBDV) strain D6948. However, vaccination often did not prevent subclinical infection in these 2-week-old progeny chicks, which often resulted in severe lymphocyte depletion in the bursa of Fabricius. Even a serum VN titre of 11 log 2 was not always sufficient to prevent severe bursal damage. Although 52/70 IBDV and vvIBDV were equally pathogenic in 2 weekold specific pathogen free chickens, significant higher maternal antibody titres were required to prevent the adverse effects of vvIBDV in comparison with 52/70 IBDV. The relation between the serological response of chickens after application of inactivated IBD vaccines and the protection of progeny chicks of vaccinated dams depended on both the virulence of the IBDV challenge strain and the IBDV strain in the vaccine. PMID- 19184921 TI - Detection of a coronavirus from turkey poults in Europe genetically related to infectious bronchitis virus of chickens. AB - Intestinal contents of 13-day-old turkey poults in Great Britain were analysed as the birds showed stunting, unevenness and lameness, with 4% mortality. At post mortem examination, the main gross features were fluid caecal and intestinal contents. Histological examination of tissues was largely unremarkable, apart from some sections that showed crypt dilation and flattened epithelia. Negative contrast electron microscopy of caecal contents revealed virus particles, which in size and morphology had the appearance of a coronavirus. RNA was extracted (turkey/UK/412/00) and used in a number of reverse transcription-polymerase chain reactions (RT-PCRs) with the oligonucleotides based on sequences derived from avian infectious bronchitis virus (IBV), a coronavirus of domestic fowl. The RT PCRs confirmed that turkey/UK/412/00 was a coronavirus and, moreover, showed that it had the same partial gene order (S-E-M-5-N-3' untranslated region) as IBV. This gene order is unlike that of any known mammalian coronavirus, which does not have a gene analogous to the gene 5 of IBV.The gene 5 of the turkey virus had two open reading frames, 5a and 5b, as in IBV and the coronaviruses isolated from turkeys in North America. The turkey/UK/412/00 also resembled IBV, but not mammalian coronaviruses, in having three open reading frames in the gene encoding E protein (gene 3). The percentage differences between the nucleotide sequences of genes 3 and 5 and the 3' untranslated region of turkey/UK/412/00 when compared with those of IBVs were similar to the differences observed when different strains of IBV were compared with each other. No sequences unique to the turkey isolates were identified. These results demonstrate, for the first time, that a coronavirus was associated with disease in turkeys outside of North America and that it is a Group 3 coronavirus, like IBV. PMID- 19184922 TI - Pathogenicity of SspI-positive infectious bursal disease virus and molecular characterization of the VP2 hypervariable region. AB - The pathogenicity of four isolates of infectious bursal disease virus (IBDV) that have restriction fragment length polymorphism patterns of very virulent IBDV (vvIBDV), based on the presence of SspI and TaqI sites in the VP2 hypervariable region, was studied in specific pathogen free chickens. Chickens inoculated with isolates 92/04, 94/B551 and 97/61 developed severe clinical signs with a high mortality ranging from 70 to 80%, whereas the 94/273 isolate caused 10% mortality. Regardless of the isolates, significant differences were noted in the bursal lesion scores and bursa:body weight ratio index in the infected groups in comparison with the control groups. However, the presence of lesions in non bursal tissues, muscles, thymus and at the junction of the proventriculus and gizzard were found only in the 92/04, 97/61 and 94/B551 isolates. Restriction fragment length polymorphism and sequence analysis of the VP2 hypervariable region indicated that all the isolates can be classified as vvIBDV based on the presence of SspI and TaqI sites at nucleotide positions 1011 and 833, respectively. In addition, all the isolates had amino acid substitutions at P222A, V256I and L294I, which are characteristic for vvIBDV isolated from different parts of the world. All the isolates except 94/273 also had a StyI site at nucleotide position 888. The absence of a StyI site in this isolate was associated with amino acid substitution at 254 from G to S. The 94/273 also had an amino acid substitution at position 270 from A to E, which is variable in the STC, Cu1 and OH strains. The presence of amino acid substitutions from G254S andA270E in SspI- and TaqI-positive vvIBDV strains is very uncommon and has not been reported previously. These amino acid variations might have caused the 94/273 to become less virulent in specific pathogen free chickens and resemble a classical virulent IBDV strain. PMID- 19184923 TI - Duck antibody responses to keyhole limpet haemocyanin, human immunoglobulin G and the trinitrophenyl hapten. Evidence of affinity maturation. AB - White Pekin ducks (three in each of four groups) received trinitrofluorobenzene (TNP) conjugated to keyhole limpet haemocyanin (KLH) or human immunoglobulin G (HIgG) either incorporated into adjuvants (complete Freund's adjuvant (CFA), then incomplete Freund's adjuvant (IFA) followed by intravenous (i.v.) injection) or by repeated i.v. injection. A rabbit received TNP-KLH with CFA, IFA, then i.v. Antibody (Ab) responses to the carrier proteins and to the TNP hapten were monitored by enzyme-linked immunosorbent assay (ELISA), optimized for use with duck Abs, and the response to TNP was also assessed in an ELISA designed to detect shifts in Ab affinity (Ka). The rabbit mounted good Ab responses to KLH and TNP, the Ka of the anti-TNP response increasing from 10(5.99) to 10(8.87) l/mol (758-fold) during the experiment. Ducks showed weak Ab responses to KLH and HIgG, with weakest responses among ducks receiving antigen by the i.v. route exclusively. The duck anti-TNP responses were vigorous in all cases. However, following i.v. administration of antigens (Ags), the anti-TNP responses were transient, being strong by 7 days but diminishing to background levels by 14 to 28 days after each inoculation. The duck Ab responses to TNP displayed affinity maturation. In ducks receiving Ags with adjuvants, Ka increased by levels comparable with or even exceeding those in the rabbit. Affinity increases were less, but nevertheless apparent, following i.v. administration of Ag. These results show that, contrary to expectation, the duck Ab response experiences affinity maturation. They also point to the transience of the duck Ab response to selected haptens/epitopes when Ag is not delivered in adjuvant. These findings have clinical implications for field responses to pathogens and vaccines. PMID- 19184924 TI - Mycoplasmas and respiratory disease in pheasants and partridges. AB - Pheasants and partridges with signs of upper respiratory disease were cultured for mycoplasmas and were also examined for Mycoplasma gallisepticum and Mycoplasma synoviae using commercial polymerase chain reaction (PCR) kits. Sixty two incidents of disease were investigated in pheasants and 12 in partridges. M. gallisepticum was detected by culture in only four and three incidents in pheasants and partridges, respectively, but with PCR a further 15 M. gallisepticum-positive incidents were detected in pheasants and another five in partridges. Several fast-growing Mycoplasma species, in particular Mycoplasma glycophilum, Mycoplasma gallinaceum and Mycoplasma pullorum, were isolated frequently and were thought to be impeding the isolation of M. gallisepticum by outgrowing it. Samples yielding M. gallisepticum isolates contained significantly fewer "contaminating" species and were exclusively from specimens submitted as whole heads rather than as swabs or as cultures from other laboratories. M. synoviae was not isolated and was detected in only one specimen by PCR. PMID- 19184925 TI - Differential attenuation of the induction by Marek's disease virus of transient paralysis and persistent neurological disease: a model for pathogenesis studies. AB - Since different biological characteristics of Marek's disease virus (MDV) are attenuated at different passage levels in cell culture, an analysis of attenuation times provides, in theory, a model for establishing the presence or absence of relationships between characteristics, thus providing a basis to link them to genetic changes in the causative virus. We have used this model to better understand the pathogenesis of the central nervous system infection as well as to evaluate the relationship of clinical neurological disease to various other parameters of MDV infection. Inoculation of 15 x7 crossbred chickens with strain 648A of very virulent plus MDV at different passage levels (between 10 and 100) showed that two neurological syndromes (transient paralysis (TP) and persistent neurological disease), were attenuated at different passage levels. While strain 648A lost the ability to induce TP between 30 and 40 passages in chicken embryo fibroblast cultures, an event closely related with all parameters of MDV infection involving viral replication (early cytolytic infection in lymphoid organs and viral replication in the feather follicle epithelium), the ability to induce persistent neurological disease was lost between 80 and 90 passages in chicken embryo fibroblasts, coincident with the loss of neoplastic lesions in peripheral nerves and other visceral organs. These data strongly suggest that transient paralysis and persistent neurological disease are unrelated and differently regulated. Moreover, comparison of brain changes induced by strain 648A at passage level 30 (TP) and at passage level 40 (no TP) also contributed to a better understanding of which brain alterations are associated with the onset of TP. The use of viruses at different passage levels with varying degrees of attenuation is presented as a useful tool for studying pathogenesis of MDV infection. PMID- 19184926 TI - Epidemiology of infectious bronchitis virus in Belgian broilers: a retrospective study, 1986 to 1995. AB - Infectious bronchitis virus (IBV) was isolated from each of 236 broiler flocks that had respiratory infection (86%), impaired growth, enteritis and/or nephritis (14%), over a 10-year period from 1986 to 1995 in Belgium. Among them, 65% of the investigated flocks had not been vaccinated against infectious bronchitis. Type specific reverse transcriptase polymerase chain reactions (RT-PCRs) were used after propagation of the isolates in embryonated eggs in order to detect and differentiate Massachusetts, D274, B1648 and 793/B types. The incidence of these types was approximately 50, 38, 11 and 1%, respectively. In 16% of cases, two or three types of IBV were detected, representing mostly combinations of Massachusetts and D274. The majority of the Massachusetts and D274 isolates (68 and 69%, respectively) were recovered from non-vaccinated flocks, confirming that such flocks are at greatest risk of infection by these types of IBV. Interestingly, the B1648 type was isolated from more vaccinated flocks (14%) than non-vaccinated flocks (7.6%). Most surprising was the very low incidence (1%) of the 793/B type, which was the dominant type in some neighbouring countries, during the period of investigation. The DNA derived by RT-PCR from 24 of the Massachusetts-type isolates from 12 vaccinated and 12 non-vaccinated flocks was sequenced and compared with the sequence of Massachusetts vaccines used in Belgium. This revealed that the sequence of four of the isolates (two from vaccinated and two from non-vaccinated flocks) was identical to that of a Massachusetts vaccine strain. Similar results were obtained for D274 isolates when compared with the sequence of D274 vaccines. These sequencing results demonstrate a co-circulation of vaccine and wild-type infectious bronchitis viruses in broilers, and are further justification for permanent monitoring of circulating strains in order to rationally modify vaccination strategies to make them appropriate to the field situation. PMID- 19184929 TI - International meetings. PMID- 19184927 TI - Protection of chickens against renal damage caused by a nephropathogenic infectious bronchitis virus. AB - The ability of the infectious bronchitis (IB) Ma5 and 4/91 live-attenuated vaccines to protect against kidney damage caused by a nephropathogenic strain of IB virus (B1648) was investigated. Protection parameters considered were gross and microscopic renal pathology, and the use of a polymerase chain reaction to detect IB RNA in kidney tissue. By each parameter, Ma5 vaccine alone provided poor protection, but 4/91 alone or the combined program both protected well. PMID- 19184930 TI - Wvpa matters. PMID- 19184931 TI - Policy regarding papers concerned with food-borne organisms. PMID- 19184932 TI - Newcastle disease virus: macromolecules and opportunities. AB - Over the past two decades, enormous advances have occurred in the structural and biological characterization of Newcastle disease virus (NDV). As a result, not only the complete sequence of the viral genome has been fully determined, but also a clearer understanding of the viral proteins and their respective roles in the life cycle has been achieved. This article reviews the progress in the molecular biology of NDV with emphasis on the new technologies. It also identifies the fundamental problems that need to be addressed and attempts to predict some research opportunities in NDV that can be realized in the near future for the diagnosis, prevention and treatment of disease(s). PMID- 19184933 TI - Production and characterization of monoclonal antibodies to fowl adenoviruses. AB - A panel of 10 monoclonal antibodies to a hypervirulent fowl adenovirus (FAdV) strain 398A was produced. Monoclonal antibodies (mAbs) were characterized for their isotype, neutralizing activity, ability to capture viral antigens, immunoblotting of viral polypeptides, competitive inhibition with chicken antisera and their reaction pattern with other FAdVs in indirect immunoperoxidase. Eight out of 10 mAbs reacted strongly in indirect immunoperoxidase staining with most of the FAdVs isolated from inclusion body hepatitis in Australia. One of these mAbs (6E1) was found to specifically react with the strains presumably characterized as hypervirulent FAdVs. IgG(2a) was the predominant sub-isotype. Two out of 10 mAbs neutralized the homologous strain of virus and six captured their target antigen onto a plastic surface. Chicken anti serum to FAdV strain 398A inhibited the reaction of the seven mAbs that bound with high affinity in a blocking competitive enzyme-linked immunosorbent assay. This panel of mAbs can be used to improve diagnostic assays, study pathogenesis and carry out strain identification. PMID- 19184934 TI - Characterization of Newcastle disease viruses isolated in Italy in 2000. AB - Thirty-two Newcastle disease virus isolates from the 2000 Italian epidemic were characterized by monoclonal antibody binding pattern and nucleotide sequencing of approximately 400 base pairs of the fusion gene. In addition, the pathogenicity of six of these isolates was assessed by means of the intracerebral pathogenicity test (ICPI). The strains tested exhibited an ICPI ranging from 1.6 to 2.0. On the basis of the monoclonal antibody binding pattern, all isolates could be classified as belonging to group C1. Both monoclonal antibody and genomic analysis revealed a very high degree of homology, indicating a common source of infection. On the basis of the phylogenetic analysis, it appears that the Italian isolates are closely related to the recent isolates from the UK, Scandinavia and South East Europe, thus suggesting the circulation of this viral strain in Europe during the past 5 years. PMID- 19184935 TI - Cloning, expression and immunogenicity of the avian pneumovirus (Colorado isolate) F protein. AB - The F protein of the Colorado isolate of avian pneumovirus (APV), expressed from a DNA plasmid, was recognized by antiserum to both A and B subgroup APVs. After two intramuscular injections of turkeys with this plasmid, a homologous antibody response was detected by enzyme-linked immunosorbent assay. This antibody also recognized subgroup A APV. However, there was no neutralization of the Colorado isolate or of subgroup A or B viruses. Although no significant clinical protection was detected following homologous challenge of poults, an anamnestic serological response was seen, suggesting that a systemic antibody response but no local mucosal immunity was induced. PMID- 19184936 TI - The influence of Escherichia coli strains from different sources and the age of broiler chickens on the development of cellulitis. AB - In two experiments, broilers were challenged with one of several field strains of Escherichia coli to determine whether the source of the E. coli and age of the bird at time of inoculation affected the development of cellulitis lesions. In the first experiment, birds inoculated at 52 days of age with E. coli of faecal, airsacculitis and cellulitis origin exhibited a cellulitis lesion incidence of 47.5, 25 and 77.5%, respectively. This study confirms earlier observations that E. coli strains isolated from cellulitis lesions express a higher propensity for producing these same lesions than other strains, including those associated with airsacculitis. In the second experiment, birds were inoculated at 4, 7, 10, 16, 28, and 52 days of age with an E. coli strain of cellulitis origin and necropsied 2 days post-infection. The resulting incidence of cellulitis ranged from 20% (day 7) to 95% (days 16 and 28), indicating that cellulitis can develop in any age of bird, although the lesions were frequently associated with other manifestations of colibacillosis (perihepatitis, pericarditis, airsacculitis) in birds challenged from 4 to 16 days of age. PMID- 19184937 TI - A comparative study of iron retention in mynahs, doves and rats. AB - Iron retention was studied in rats (Rattus norvegicus), doves (Streptopelia d. decaocto) and two species of mynahs (Acridotheres t. tristis and Gracula r. religiosa) fed two different pelleted diets (88.5 and 567.9mg Fe/kg diet). The doves and rats served as species that are not susceptible to iron storage, whereas the mynahs are known to develop iron overload frequently. The retention was calculated after measuring the uptake and elimination of a single dose of radioactive iron ((59)Fe) using whole-body counting. It was hypothesized that the mynahs would retain more iron than the rats and doves, and that after dietary iron challenge the mynahs would downregulate iron retention less effectively. It is concluded that mynahs have much higher iron uptake and retention than doves, but a similar uptake to that in rats. The four studied species are able to downregulate iron retention, the doves being the most efficient. It is suggested that at least part of the susceptibility to iron overload in mynahs is related to a high iron absorption from the intestines regardless of body iron stores, which is comparable with the situation of hereditary haemochromatosis in man. PMID- 19184938 TI - Characterization and experimental reproduction of peripheral neuropathy in White Leghorn chickens. AB - A clinical neurological syndrome termed peripheral neuropathy (PN) that resembles Marek's disease (MD) occurred at low frequency in a commercial layer strain for several years. Study of chickens from six field cases showed that the PN syndrome could be distinguished pathologically from MD on the basis of several factors, including onset as early as 6 weeks, presence of B-type but not A-type lesions in peripheral nerves, and absence of visceral lymphomas. Serotype 1 MD virus could not be isolated from blood from any chicken or demonstrated in tissues by histochemistry or polymerase chain reaction assays. Moreover, the syndrome was not prevented by MD vaccination, either in the field or in laboratory trials. PN was induced in 3 to 54%of commercial line chickens inoculated at 1 or 6 days of age with whole blood or buffy coat cells from clinically affected donor chickens. Sonicated cells also induced PN, but plasma was ineffective. Chickens did not develop PN if reared in isolators without cellular transfer or when vaccinated solely against MD. However, PN was observed in 9% of 57 B*2/*19 commercial chickens reared in isolators following vaccination against MD, infectious bursal disease, Newcastle disease and infectious bronchitis, suggesting that common vaccines may predispose chickens to PN. The data confirmed a strong influence of the major histocompatibility complex (B-complex) on both naturally occurring and experimentally induced PN with the B*19 haplotype conferring susceptibility compared with other alleles. It is postulated that PN may represent an autoimmune reaction to nerve tissue that may result from response to a combination of common vaccines. These studies confirmed that PN is distinct from MD, provided criteria for its differential diagnosis, identified strategies for its control, and established a model for its experimental induction. PMID- 19184939 TI - Sequence analysis of the VP2 gene hypervariable region of infectious bursal disease viruses from India. AB - Attempts have been made to characterize infectious bursal disease virus (IBDV) isolates collected from different parts of India during 1993 to 1999. Phylogenetic analysis was performed on a sequence generated by cycle sequencing comprising the variable region of the VP2 gene of 14 isolates. Indian IBDV isolates had divergence of 0.2 to 4.3% at nucleotide and 0 to 2.2% at amino acid levels among themselves. Nine nucleotide changes were found in Indian IBDV field isolates, resulting in the four specific amino acid changes at 222P-A, 256V-I, 294L-I and 299N-S, reported regularly in very virulent isolates. One of the Indian IBDV isolates, UP1/99, had change D to N at position 212 in the first hydrophilic region. The serine-rich heptapeptide sequence 'SWSASGS' was conserved in all the isolates. Phylogenetically, all Indian field isolates were found to be closely related to very virulent IBDV isolates from Europe, Japan, China and Israel. PMID- 19184940 TI - Safety and efficacy of water-in-oil-in-water emulsion vaccines containing Newcastle disease virus haemagglutinin-neuraminidase glycoprotein. AB - Subunit vaccines containing haemagglutinin-neuraminidase (HN) glycoprotein of Newcastle disease virus (NDV), formulated as water-in-oil-in-water (W/O/W) emulsions, were prepared. First, the suitable constituents of a W/O/W emulsion adjuvant were investigated with polyvalent vaccines using NDV, infectious bronchitis virus and Haemophilus paragallinarum. The W/O/W emulsion adjuvant, composed of the antigen in phosphate-buffered saline (PBS), liquid paraffin, squalene, diglyceryl monooleate, polysorbate 80 and PBS in a 30:25:10:5:2:28 ratio, induced a good antibody response with less adverse local reactions. HN protein of NDV was expressed by an improved baculovirus expression vector, a hybrid nucleopolyhedrovirus (HyNPV) between Autographa californica NPV and Bombyx mori NPV,and was prepared from silkworm pupae infected with the recombinant baculovirus, HyNPV-HN. Then, the W/O/W emulsion vaccine containing HN protein was prepared using the aforementioned constituents. Chickens showed 100, 100 and 80% protection against challenge exposure to virulent NDV at 4 weeks after vaccination with W/O/W emulsion vaccines containing 30, 6 and 3% of HyHPV-HN infected pupae, respectively. The vaccines containing HN protein did not induce adverse local reactions at the site of injection. The subunit vaccine for NDV containing HN protein expressed in the recombinant baculovirus-infected pupae, formulated as a W/O/W emulsion vaccine composed of the antigen in PBS, liquid paraffin, squalene, diglyceryl monooleate, polysorbate 80 and PBS in a 30:25:10:5:2:28 ratio, was therefore found to be safe and effective. PMID- 19184941 TI - Chronic pulmonary interstitial fibrosis in Amazon parrots. AB - In the Netherlands, a syndrome affecting several species of older Amazon parrots (Amazona sp.) has been described. This syndrome was characterized as a chronic respiratory disease resulting in exercise intolerance. Pathological examination revealed loss of functional lung tissue, pulmonary interstitial fibrosis, and right heart failure. Haematology revealed an elevated packed cell volume as a result of an increase in erythrocyte size and an increased haemoglobin mass per erythrocyte. In two patients, hypoxia and hypercapnia were demonstrated. The aetiology of this syndrome is currently not known. The microscopic lesions resemble those found in Diffuse Interstitial Fibrosis in humans. PMID- 19184942 TI - Influence of vaccine deposition site on post-vaccinal viraemia and vaccine efficacy in broiler chickens following in ovo vaccination against Marek's disease. AB - In ovo vaccination against Marek's disease is a widely used technology in the broiler industry.A series of experiments was carried out to determine the site of vaccine deposition in the egg during automated in ovo vaccination, and the effect of vaccine deposition site and dose on vaccine responses following vaccination with cell-associated herpesvirus of turkeys in commercial broiler chickens. Vaccine deposition site following automated in ovo vaccination was principally influenced by the age of embryo, with egg size having a smaller effect. The frequency of vaccine deposition inside the embryo body increased as incubation progressed from day 17.5 to 19.5. In experiments using manual vaccine deposition intra-embryonically (IE) or extra-embryonically (EE) at day 18.5, EE vaccine deposition resulted in a significantly delayed development of post-vaccinal viraemia relative to both IE vaccination and subcutaneous vaccination at hatch. There were no effects of vaccine dose (2000, 4000 or 8000 plaque forming units) on the timing of post-vaccinal viraemia. The timing of post-vaccinal viraemia was found to be a good indicator of the level of protection provided by the vaccine against challenge with earlier viraemia associated with better protection. IE vaccine deposition induced significantly greater protection than EE deposition against challenge with a virulent strain of Marek's disease virus. IE deposition consistently produced a high level of protection (68 to 84%) irrespective of vaccine dose or challenge day, while EE vaccine deposition produced no or low levels of protection (0 to 27%) depending on the vaccine dose and day of challenge. The growth of challenged chickens was also affected by site of vaccine deposition, with significantly higher live weights at day 56 of age in IE compared with EE vaccinated groups. These data suggest that the site of vaccine deposition within the embryo is an important determinant of the success of in ovo vaccination. PMID- 19184943 TI - Sequence analysis of nephropathogenic infectious bronchitis virus strains of the Massachusetts genotype in Beijing. AB - The first 440 nucleotides of the S1 part of the spike protein (S) gene of three nephropathogenic infection bronchitis virus (IBV) strains (BJ1, BJ2 and BJ3), isolated from three rural chicken farms of Beijing with outbreaks of avian infectious bronchitis, were sequenced after reverse transcriptasenested polymerase chain reaction amplification. The sequence data were analyzed and compared with the standard IBV strains M41, Beaudette, vaccine strain H120 and Chinese vaccine strain D41. The three IBV isolates and strain M41 showed more than 99.1 and 97.8% identity in nucleotide sequence and deduced amino acid sequence, respectively, suggesting that the three nephropathogenic IBV strains belonged to the Massachusetts genotype of IBV. PMID- 19184944 TI - Validation of five commercially available ELISAs for the detection of antibodies against infectious bursal disease virus (serotype 1). AB - In this study, the results are reported from a validation study of five commercially available enzymelinked immunosorbent assays (ELISAs) for the detection of antibodies against infectious bursal disease virus (IBDV), serotype 1. The specificity of the ELISAs varied from 63.8 to 100%. All ELISAs reached a sensitivity of 100% on sera between 14 and 21 days post-vaccination (d.p.v.) with two classical vaccines and a Delaware variant-E virus. Overall, most birds became positive between 8 and 11 d.p.v. As expected, the ELISA with the lowest specificity showed the highest sensitivity at 5 d.p.v. When the decrease in maternally derived antibodies against IBDV was measured, a highly significant correlation (P < 0.001) was found for all ELISAs and the virus neutralization test (VNT). PMID- 19184945 TI - Differentiation of psittacine herpesvirus field isolates by restriction endonuclease analysis. AB - Thirty-one viruses were isolated from 15 different psittacine species. They were all identified as herpesviruses on the basis of chloroform sensitivity, inhibition of replication in cell cultures by 5-iodo-2'-deoxyuridine and morphology by electron microscopy. DNA from all 31 field isolates of psittacine herpesviruses (PsHV) was analysed by cleavage with restriction endonucleases EcoRI, PstI, and BglII. Using this technique, 12 different restriction patterns were recognizable. In previous work, we have differentiated PsHV strains according to their serological interrelationships. Restriction endonuclease profiles confirmed the results of conventional serogrouping but additional differences were observed among the five established serological subtypes. Using this method, it was possible to demonstrate a spontaneous reinfection of parrots in an aviary with a serologically and genetically different PsHV strain 4 years after the first outbreak of Pacheco's parrot disease (PPD). The results of this study reaffirm that antigenetically and genetically different PsHV exist and can cause PPD. PMID- 19184946 TI - Cathaemasia hians (Trematoda: Cathaemasiidae) infecting black stork nestlings (Ciconia nigra) from central Spain. AB - The trematode Cathaemasia hians is recorded for the first time in nestling Black storks, and for the first time in Black storks in Spain. The parasite was found in the upper part of the oesophagus of Black storks (Ciconia nigra). Presence of this parasite in three nestlings raises the possibility of completion of the parasite's life cycle in Europe. In addition, a sub-adult bird recuperating from bone fractures was also found to be infected. C. hians is apparently absent from the more common white stork (Ciconia ciconia) from the same area. The different feeding behaviour of the two species of storks may account for this. PMID- 19184949 TI - International meetings. PMID- 19184947 TI - Peripheral neuritis in psittacine birds with proventricular dilatation disease. AB - Necropsies were performed on 14 psittacine birds of various species suspected to have proventricular dilatation disease (PDD). Eight of the birds exhibited neurological signs (seizures, ataxia, tremors and uncoordinated movements) and digestive tract signs (crop stasis, regurgitation, inappetance and presence of undigested food in the faeces). At necropsy, the birds had pectoral muscle atrophy, proventricular and ventricular distention, thinning of the gizzard wall, and duodenal dilation. In addition, five birds had a transparent fluid (0.2 to 1.0 ml) in the subarachnoidal space of the brain, and one bird had dilatation of the right ventricle of the heart. The histological lesions differed from earlier reports of PDD in that peripheral (sciatic, brachial and vagal) neuritis was seen in addition to myenteric ganglioneuritis, myocarditis, adrenalitis, myelitis and encephalitis. PMID- 19184950 TI - Wvpa matters. PMID- 19184952 TI - Innovation and discovery: the application of nucleic acid-based technology to avian virus detection and characterization. AB - Polymerase chain reaction (PCR)-based approaches to the detection, differentiation and characterization of avian pathogens continue to be developed and refined. The PCRs, or reverse transcriptase-PCRs, may be general, designed to detect all or most variants of a pathogen, or to be serotype, genotype or pathotype specific. Progress is being made with respect to making nucleic acid approaches more suitable for use in diagnostic laboratories. Robotic workstations are now available for extraction of nucleic acid from many samples in a short time, for routine diagnosis. Following general PCR, the DNA products are commonly analyzed by restriction endonuclease mapping (restriction fragment length polymorphism), using a small number of restriction endonucleases, based on a large body of sequence data. Increasingly, however, nucleotide sequencing is being used to analyze the DNA product, in part due to the expanding use of non radioactive sequencing methods that are safe and enable high throughout. In this review, I highlight some recent developments with many avian viruses: Newcastle disease virus; circoviruses in canary and pigeon; infectious bursal disease virus (Gumboro disease virus); avian adenoviruses, including Angara disease/infectious hydropericardium virus, haemorrhagic enteritis virus of turkeys, and egg drop syndrome virus; avian herpesviruses, including infectious laryngotracheitis virus, duck plague virus, psittacine herpesvirus (Pacheco's parrot disease virus), Marek's disease virus and herpesvirus of turkeys; avian leukosis virus (associated with lymphoid leukosis or myeloid leukosis, and egg transmission); avian pneumoviruses (turkey rhinotracheitis virus); avian coronaviruses, including infectious bronchitis virus, turkey coronavirus and pheasant coronavirus; astrovirus, in the context of poult enteritis and mortality syndrome, and avian nephritis virus; and avian encephalomyelitis virus, a picornavirus related to hepatitis A virus. PMID- 19184953 TI - Diphacinone and zinc phosphide toxicity in a flock of Peafowl. AB - Toxicity probably due to a combination of diphacinone and zinc phosphide was diagnosed in a flock of peafowl, in which 35 birds in a flock of 80 died over a span of 10 days without any apparent clinical signs. Chickens and guinea fowl, 30 each on the same premises, were not affected. Plastic tubes containing diphacinone and zinc phosphide were used on the premises to control ground squirrels. Most of the six dead peafowl, which ranged in age from 6 months to 4 years, had an accumulation of serosanguinous fluid in the abdominal cavity, semi clotted blood over the liver lobes, increased pericardial fluid, and enlarged and pale kidneys. Pellets of diphacinone and zinc phosphide were found in the crop and gizzard contents from most of the birds. Microscopically, most of the birds had mild to moderate centrolobular degeneration of hepatocytes and multifocal degeneration of myofibres in the heart with infiltration by a few mononuclear cells. Acute nephrosis and mucosal oedema in the oesophagus and crop were also observed. Toxicological analysis of the crop and gizzard contents revealed the presence of diphacinone and phosphine gas, and analysis of the crop contents from two birds for heavy metals revealed zinc levels of up to 6600 parts/10 6 . It was suspected that only the peafowl and not the chickens and guinea fowl were affected because peafowl, with their longer necks, were able to reach into the plastic tubes and eat the pellets. PMID- 19184954 TI - A diagnostic study on columbid circovirus infection. AB - A systematic study was performed to examine the frequency of columbid circovirus (CoCV) infection in diseased young pigeons submitted for necropsy and its relevance to pigeon health. Existing diagnostic methods were compared. Among 176 diseased young pigeons examined, CoCV infection was detected in 83 cases using negative contrast electron microscopy. Histopathological examination allowed a clear diagnosis in only 42 pigeons. Therefore, a polymerase chain reaction assay and an in situ hybridization test were developed as additional diagnostic tools. CoCV is by far the most frequently detected infectious agent in diseased young pigeons. Infected pigeons reveal a broad range of concurrent infections. Pathological findings suggest an immunosuppressive effect of CoCV. PMID- 19184955 TI - The antigenicity of the chicken anaemia virus protein VP3 (Apoptin). AB - Chicken anaemia virus protein VP3 (Apoptin) was cloned and expressed as a recombinant protein and evaluated for its suitability as a serodiagnostic reagent. VP3 was expressed as a fusion protein either with glutathione S transferase or with a six-histidine tag. Both recombinant proteins reacted specifically with anti-VP3 monoclonal antibodies and with serum from vaccinated chickens by Western blot and by enzyme-linked immunosorbent assay (ELISA). However, when testing sera from birds of different ages and genetic backgrounds, high non-specific reactions were evident and false positives were observed, especially in older birds. This suggests that VP3 is poorly immunogenic during infection and low antibody concentrations are masked by non-specific reactions. Thus, VP3 is not suitable for use as antigen in ELISAs. PMID- 19184956 TI - Marek's disease in broiler chickens: effect of route of infection and herpesvirus of turkey-vaccination status on detection of virus from blood or spleen by polymerase chain reaction, and on weights of birds, bursa and spleen. AB - The polymerase chain reaction (PCR) has recently emerged as an additional tool for the monitoring and diagnosis of Marek's disease. We investigated a number of factors that may influence the interpretation of PCR results in commercial broiler chickens including the effects of route of infection and herpesvirus of turkeys (HVT)-vaccination status. We also investigated the suitability of peripheral blood lymphocytes (PBL) and spleen as tissues for Marek's disease virus (MDV) detection. HVT-vaccinated and unvaccinated commercial broiler chickens were challenged or not challenged with virulent MDV either by intraperitoneal injection or inhalation of feather dust containing the virus. Blood and spleen samples were collected at weekly intervals to day 35 post infection for PCR of spleen or PBL. Live weight and lymphoid organ weights were also measured. Spleen and PBL were found to provide similar sensitivity of detection of MDV with a small advantage in favour of spleen. In terms of the timing of detection of MDV, intraperitoneal challenge broadly mimicked natural challenge via inhalation, although infection of birds by inhalation of infective feather dust resulted in slightly later but more complete detection of MDV in challenged birds. Vaccination with HVT delayed the detection of MDV by approximately 10 to 14 days and did not protect against the reduced growth observed in challenged chickens at day 35 post-challenge. PMID- 19184957 TI - Field studies on the association between amyloid arthropathy and Mycoplasma synoviae infection, and experimental reproduction of the condition in brown layers. AB - Approximately 20% of outbreaks of amyloid arthropathy in The Netherlands were earlier attributed to Enterococcus faecalis but there was no explanation for the remaining cases. In a further study, material from the joints of 10 birds from each of 10 affected brown layer flocks and two broiler flocks was cultured in several bacteriological media. E. faecalis was isolated from one layer flock but Mycoplasma synoviae was recovered from purulent joints in six of the layer flocks and both broiler flocks. Brown layers were then infected experimentally with one of the M. synoviae isolates to assess its arthropathic and amyloidogenic potential. Using Congo red staining, articular amyloid deposits were seen 12 weeks after intra-articular or intravenous inoculation in almost all birds. After intra-articular inoculation of the left knee joint the contralateral hock tendon sheath tended to be affected, while hock and foot joints were mainly affected after intravenous inoculation. Amyloid deposits were seen in livers and spleens of many birds. This is the first demonstration of arthropathic strains of M. synoviae with an association with amyloid arthropathy, while for one isolate its amyloidogenic potential was shown experimentally. PMID- 19184958 TI - Reduction of horizontal transmission of avian leukosis virus subgroup J in broiler breeder chickens hatched and reared in small groups. AB - Transmission of avian leukosis virus, subgroup J (ALV-J), from donor chickens inoculated as embryos to simulate congenital infection to uninfected hatchmates was studied in two strains of commercial broiler breeder chickens. Chicks of two commercial lines free of ALV-J became infected when hatched (1/2 lots positive) or reared (8/8 lots positive) in direct physical contact with ALV-J-infected donors. Infection also occurred when chicks were exposed in the hatchery to ALV-J infected donors by cloacal swab transfer (2/2 lots positive), needle transfer during subcutaneous inoculation (2/2 lots positive), or ingestion of infected meconium (2/2 lots positive). However, transmission was delayed or prevented by wire partitions in the hatcher and rearing of small groups in cubicles, and rarely (1/10 lots positive) resulted from short-term direct or indirect contact. In a simulated field test, a flock of 503 broiler breeder chickens with an initial embryo infection rate of 4.6% was hatched and reared as 48 small groups to 4 weeks of age. Groups were tested at hatch and at 3 weeks, and 14 infected groups were eliminated. This flock tested negative for ALV-J infection from 4 to 32 weeks and did not transmit infection to progeny or develop tumours. A control group of 377 chickens with a similar initial infection rate was hatched and reared as a single group. This control flock transmitted virus to 5.7% of its progeny and about 5% of the hens developed tumours. The small-group hatching and rearing practices employed in these studies allowed for the accurate identification and removal of groups containing chickens infected prior to hatching and prevented horizontal transmission of ALV-J between uninfected and infected groups for at least 4 weeks. More importantly, application of these procedures successfully eradicated ALV-J in a single generation under laboratory conditions. This suggests that similar procedures could be a valuable adjunct to virus eradication programmes in the field. PMID- 19184959 TI - Polymerase chain reaction combined with restriction enzyme analysis for detection and differentiation of fowl adenoviruses. AB - A polymerase chain reaction combined with restriction enzyme analysis was developed for detection and differentiation of all 12 fowl adenovirus (FAdV) serotypes representing the five fowl adenovirus (A to E) species. For primer design, the published sequences of the hexon proteins of FAdV1, FAdV8 and FAdV9 were aligned and conserved regions in the two pedestal regions adjacent to the L1 loop region were determined. A primer pair (hexon A/hexon B) was constructed and was shown to amplify approximately 900 bp of the hexon gene including the L1 loop region. An amplification product was detected using supernatant of infected cell cultures from all FAdV1 to FAdV12 reference strains used in our study. The sequence and the restriction patterns of the hexon A/B fragments of the 12 FAdV strains were determined and compared. The successive use of four different endonucleases allowed the complete differentiation of the reference FAdV strains. Twenty-six fowl adenoviruses isolated during our routine virological diagnosis activities could all be amplified using hexon A/hexon B primers. Restriction analysis results showed that 8/26 adenovirus strains contained two different FAdV types. FAdV4, FAdV12, FAdV1, FAdV5 and FAdV6 were the most frequently isolated. PMID- 19184960 TI - Group I avian adenovirus and avian adeno-associated virus in turkey poults with inclusion body hepatitis. AB - Adenoviral inclusion body hepatitis (IBH) is rare in turkeys. Avian adenovirus group I and avian adeno-associated virus were isolated from the liver and pooled intestinal samples from 4-week-old turkey poults on two different ranches experiencing increased mortality. Grossly, a few birds from each ranch had a slightly enlarged liver with white foci of necrosis randomly scattered throughout. Microscopically, there was coagulative necrosis of hepatocytes with infiltration of a mixed population of inflammatory cells composed of lymphocytes, plasma cells, heterophils, and macrophages. In these livers, there were numerous basophilic intranuclear inclusion bodies in the hepatocytes. Transmission electron microscopy of the liver revealed 70 to 75 nm viral particles with icosahedral morphology consistent with adenovirus scattered throughout the nucleus of hepatocytes. All of the birds were serologically negative for haemorrhagic enteritis virus infection. Of some 5000 submissions over a 12-year period of turkey poults aged between 1 day and 10 weeks, only two single birds within two submissions had IBH. PMID- 19184961 TI - Amino acid changes in the variable region of VP2 in three infectious bursal disease viruses with different virulence, originating from a common ancestor. AB - An infectious bursal disease virus (IBDV), Hyd(C), from an outbreak was isolated and plaque-purified in BGM-70 cells. From the moderately virulent plaque-purified virus, two IBDVs with relatively high and low virulence were obtained by passaging the virus in specific pathogen free chickens and BGM-70 cells 10 and seven times, respectively. Comparison of amino acid sequences of the VP2 variable region of these viruses revealed that three amino acids at positions 279, 284 and 300 (Asp, Thr and Glu, respectively, in the plaque-purified virus) were changed. In in vitro- passaged virus, amino acid residues 279, 284 and 300 were Asn, Thr and Glu, whereas these were Asp, Ala and Gln in the in vivo -passaged virus. Change of residue 284 (Thr -->Ala) had a critical role in cell culture infectivity, whereas the change in residue 279 (Asp -->Asn) was associated with attenuation of the virus. No correlation could be observed between amino acid changes at position 300 and virulence or cell culture infectivity. Moreover, residue 330 (Arg) in heptapeptide motif SWSAR 330 GS was not found to be associated with the cell culture infectivity or virulence. PMID- 19184962 TI - Avian leukosis virus subtype J in ovo -infected specific pathogen free broilers harbour the virus in their feathers and show feather abnormalities. AB - In the present paper, abnormal feather development associated with an in ovo avian leukosis virus (ALV) subgroup J infection in specific pathogen free broilers is reported. A description of gross lesions is given, together with the results of light microscopic, immunohistochemical and ultrastructural studies to further elucidate the characteristics of the feather pathology and to assess the presence of ALV subtype J in broiler feathers. The feather lesions found, which were mainly restricted to primary and secondary remiges, were thinness and increased transparency of the calamus and sparseness of the vane. In a few cases, feather growth retardation was present, while the vane remained ensheathed. Light microscopy did not reveal abnormalities, although immunohistochemically prominent anti-p27 staining was found in the central area of the feather pulp between cells forming the barbular epithelium. Transmission electron microscopy showed the presence of retroviral particles in various structures of the feather (epidermal collar and intermediate layer of feather epidermis), most virus being present in the same area that stained most prominently by immunohistochemistry and where apparently intra-epithelial gaps were seen. ALV was isolated from the upper two thirds of primary remiges of almost all inoculated birds and 40% of contact birds. Polymerase chain reaction of cell cultures showed that the virus isolates belonged to subgroup J. This study shows that subtype J virus can be massively present in feather tissue of infected birds, which could have consequences for its transmission and diagnosis. PMID- 19184965 TI - Wvpa matters. PMID- 19184964 TI - International meetings. PMID- 19184966 TI - Phenylpropanoid and lignan derivatives from Antiaris toxicaria and their effects on proliferation and differentiation of an osteoblast-like cell line. AB - Antiarisin A and B (1 and 2), together with seventeen known compounds (3-19), were isolated from the EtOAc extract of the stem of Antiaris toxicaria Lesch. Their structures were determined on the basis of spectroscopic analyses and chemical methods. Most of the compounds were reported for the first time from the Antiaris genus and firstly studied for their proliferative and differentiative activity on osteoblast-like cells. Screening results indicated that, at the concentration of 10(-8) M, benzofuran lignans 5, 6, 11 and 13 could significantly stimulate the proliferation of UMR106 cells, while 8, 9, 11, 14, 15 and 17 could enhance ALP (alkaline phosphatase) activity. PMID- 19184967 TI - Lonicera hypoglauca inhibits xanthine oxidase and reduces serum uric acid in mice. AB - Xanthine oxidase (XOD) catalyzes the oxidation of hypoxanthine to xanthine and then to uric acid, and is a key enzyme in the pathogenesis of hyperuricemia. The ability of extracts of Lonicera hypoglauca (Caprifoliaceae) to inhibit XOD was investigated in this study. An ethanol extract (LH-crude) of the leaves of L. hypoglauca and its derived EtOAc soluble sub-fractions (LH-EA) significantly inhibited XOD activity, with IC50 values for LH-crude and LH-EA of 48.8 and 35.2 microg/mL. Moreover, LH-EA reduced serum urate levels IN VIVO in a potassium oxonate-induced hyperuricemic mouse model, by 70.1% and 93.7% of the hyperuricemic untreated group at doses of 300 and 500 mg/kg of LH-EA, respectively. Finally, we used bioactivity-guided fractionation to isolate a new bisflavonoid, loniceraflavone, which showed significant inhibition of XOD (IC50=0.85 microg/mL). These results suggest that L. hypoglauca and its extracts may have a considerable potential for development as an anti-hyperuricemia agent for clinical application. PMID- 19184968 TI - Antinociceptive effect and GC/MS analysis of Rosmarinus officinalis L. essential oil from its aerial parts. AB - The rationale of this investigation was to examine the antinociceptive properties of the essential oil obtained from Rosmarinus officinalis aerial parts, using a rat model of arthritic pain. The essential oil (100, 300 and 600 mg/kg, I. P.) produced a dose-dependent antinociceptive effect, manifested as a significant reduction in the dysfunction in the pain-induced functional impairment model in the rat (PIFIR model), mainly at high doses. Chemical constituents of the essential oil were further analyzed by gas chromatography-mass spectrometry (GC/MS). The major compounds in the essential oil were alpha-pinene (14.10 %), camphene (11.47 %), beta-pinene (12.02 %), myrcene (3.31 %), alpha-phellandrene (7.87 %), eucalyptol (8.58 %), 2-bornanone (3.42 %), camphor (8.75 %), isoborneol (3.48 %), borneol (4.85 %) and borneol acetate (6.49 %). The antinociceptive effects of R. officinalis essential oil were tested in combination with 0.12 mg/kg WAY100635, s. c. (an antagonist of 5-HT(1A) receptors) or 1 mg/kg naloxone, i. p. (an antagonist of endogenous opioids receptors), demonstrating in both cases an inhibition of the antinociceptive response. This study suggests an involvement, at least in part, of the serotonergic system via 5-HT(1A) receptors and endogenous opioids in the antinociceptive effect of R. officinalis essential oil in the PIFIR model. PMID- 19184970 TI - A new specific BACE-1 inhibitor from the stembark extract of Vitis vinifera. AB - A new resveratrol dimer, (+)-vitisinol E (1) which demonstrated inhibitory activity on BACE-1 (beta-site APP-cleaving enzyme 1) in vitro, was isolated from the stembark extract of Vitis vinifera (Vitaceae) together with four known resveratrol oligomers, (+)-epsilon-viniferin (2), (+)-ampelopsin A (3), (+) vitisin A (4) and (-)-vitisin B (5). The chemical structure of 1 was established by MR spectroscopic analyses, including HMBC. All isolated resveratrol derivatives (1-5) demonstrated significant inhibition on baculovirus-expressed BACE-1 in a dose-dependent manner, which was assessed with the FRET assay using Rh-EVNLDAEFK as a substrate in vitro. PMID- 19184969 TI - Dammarane-type triterpene glycosides from Oncoba manii active against methicillin resistant Staphylococcus aureus. AB - Drug-resistant bacteria are becoming more prevalent both in the community and in hospitals. In a search for new antibiotic leads, we used a high-throughput natural products chemistry approach to isolate one new (1) and two known (2, 3) dammarane-type triterpenes with mass-limited material from the African plant Oncoba manii. The new compound was determined by spectroscopic methods to be 1beta,2alpha,3beta,20(R)-tetrahydroxydammar-24-ene 3-O-alpha-L-rhamnopyranosyl-(1 --> 2)-beta-D-glucopyranoside. Compounds 1 and 2 inhibited the growth of methicillin-resistant Staphylococcus aureus (MRSA). PMID- 19184971 TI - Disorders of prosencephalic development. AB - Abnormal ventral induction may result in disorders of formation, cleavage, and midline development of prosencephalic structures. Holoprosencephaly is a developmental field defect of impaired cleavage of prosencephalon. The most widely accepted classification of holoprosencephaly recognizes three major varieties: the alobar, semilobar and lobar types, according to the severity of the malformation. The brain malformations, characterized by the fusion of the cerebral hemisphere along the midline are commonly associated with facial anomalies. Corpus callosum agenesis and septo-optic dysplasia are disorders of prosencephalic midline development, and usually have less severe presentations but still, affected subjects may suffer from neurodevelopmental retardation, and/or endocrinologic and visual disorders. In this article we report an up-to date of pathogenesis, prenatal sonographic findings, differential diagnosis and prognosis of the aforementioned anomalies. PMID- 19184972 TI - The significance of fetal ventriculomegaly: etiology, short- and long-term outcomes. AB - Fetal cerebral ventriculomegaly (VM) is diagnosed when the width of one or both ventricles, measured at the level of the glomus of the choroid plexus (atrium), is > or = 10 mm. VM can result from different processes: abnormal turnover of the cerebrospinal fluid (CSF), neuronal migration disorders, and destructive processes. In a high percentage of cases, it is associated with structural malformations of the central nervous system (CNS), but also of other organs and systems. The rate of associated malformations is higher (> or =60%) in severe VM (>15 mm) and lower (10-50%) in cases of borderline VM (10-15 mm). When malformations are not present, aneuploidies are found in 3-15% of borderline VM; the percentage is lower in severe VM. The neurodevelopmental outcome of isolated VM is normal in > 90% of cases if the measurement of ventricular width is between 10 and 12 mm; it is less favorable when the measurement is > 12 mm. PMID- 19184973 TI - Proteome profiling of aging in mouse models: differential expression of proteins involved in metabolism, transport, and stress response in kidney. AB - Aging is a time-dependent complex biological phenomenon observed in various organs and organelles of all living organisms. To understand the molecular mechanism of age-associated functional loss in aging kidneys, we have analyzed the expression of proteins in the kidneys of young (19-22 wk) and old (24 months) C57/BL6 male mice using 2-DE followed by LC-MS/MS. We found that expression levels of 49 proteins were upregulated (p < or = 0.05), while that of only ten proteins were downregulated (p < or = 0.05) due to aging. The proteins identified belong to three broad functional categories: (i) metabolism (e.g., aldehyde dehydrogenase family, ATP synthase beta-subunit, malate dehydrogenase, NADH dehydrogenase (ubiquinone), hydroxy acid oxidase 2), (ii) transport (e.g., transferrin), and (iii) chaperone/stress response (e.g., Ig-binding protein, low density lipoprotein receptor-related protein associated protein 1, selenium binding proteins (SBPs)). Some proteins with unknown functions were also identified as being differentially expressed. ATP synthase beta subunit, transferrin, fumarate hydratase, SBPs, and albumin are present in multiple forms, possibly arising due to proteolysis or PTMs. The above functional categories suggest specific mechanisms and pathways for age-related kidney degeneration. PMID- 19184974 TI - Composite measures for hospital quality using quality-adjusted life years. AB - Developing clinically meaningful summary measures of health-care quality is key to inferring quality of care. Current summary measures use a number of different approaches to weight their individual measures but rarely use weights based on clinical 'importance'. Such an approach would help to focus quality improvement efforts on areas likely to have the largest impact on health outcomes. Using coronary artery bypass graft (CABG) surgery as a case study, we weight and combine 11 process, complication, and survival measures to summarize differences in quality-adjusted life expectancy 1 year following surgery for a sample of hospitals. We use a fully Bayesian analysis to estimate 1-year survival outcomes using a hierarchical exponential survival model. We then estimate the expected utility of the year following surgery for each patient using complication probabilities fitted from hierarchical models and utility values from the literature. We estimate quality-adjusted life years (QALYs) for each hospital as the utility-weighted average 1-year survival probability and then estimate 'incremental QALYs' by taking the difference in QALYs for each hospital relative to a comparison group that reflects the average performance of all hospitals in the state. We illustrate our framework by estimating incremental QALYs for 14 hospitals performing CABG surgery in Massachusetts in 2003 and find that a composite measure based on QALYs can change the classification of quality outliers relative to conventional mortality measures. PMID- 19184975 TI - Mediation analysis with principal stratification. AB - In assessing the mechanism of treatment efficacy in randomized clinical trials, investigators often perform mediation analyses by analyzing if the significant intent-to-treat treatment effect on outcome occurs through or around a third intermediate or mediating variable: indirect and direct effects, respectively. Standard mediation analyses assume sequential ignorability, i.e. conditional on covariates the intermediate or mediating factor is randomly assigned, as is the treatment in a randomized clinical trial. This research focuses on the application of the principal stratification (PS) approach for estimating the direct effect of a randomized treatment but without the standard sequential ignorability assumption. This approach is used to estimate the direct effect of treatment as a difference between expectations of potential outcomes within latent subgroups of participants for whom the intermediate variable behavior would be constant, regardless of the randomized treatment assignment. Using a Bayesian estimation procedure, we also assess the sensitivity of results based on the PS approach to heterogeneity of the variances among these principal strata. We assess this approach with simulations and apply it to two psychiatric examples. Both examples and the simulations indicated robustness of our findings to the homogeneous variance assumption. However, simulations showed that the magnitude of treatment effects derived under the PS approach were sensitive to model mis-specification. PMID- 19184977 TI - A finite mixture survival model to characterize risk groups of neuroblastoma. AB - Neuroblastoma is a childhood cancer with patients experiencing heterogeneous survival outcomes despite aggressive treatment. Disease outcomes range from early death to spontaneous regression of the tumor followed by cure. Owing to this heterogeneity, it is of interest to identify patients with similar types of neuroblastoma so that specific types of treatment can be developed. Oncologists are especially interested in identifying patients who will be cured so that the minimum amount of a potentially toxic treatment can be given to this group of patients. We analyze a large cohort of neuroblastoma patients and develop a finite mixture model that uses covariates to predict the probability of being in a cure group or other (one or more) risk groups. A prediction method is developed that uses the estimated probabilities to assign a patient to different risk groups. The robustness of the model and the prediction method is examined via simulation by looking at misclassification rates under misspecified models. PMID- 19184978 TI - Cell proteomic footprint. AB - The authentication of mammalian cell cultures and their subpopulations is of great demand in biotechnology and cell therapy. However, current techniques are either not efficient or can be very complex and expensive. Here we report a simple and straightforward approach for authentication of biological cells and their subpopulations with high speed, high throughput, low sample cost, and high sensitivity. We discovered that cell cultures treated with protease under soft, 'non-killing' conditions release fragments of cell surface proteins, whose composition is a strong characteristic of the cells. Mass spectrometric analysis of the released fragments allows a direct comparison of the produced mass spectrum with the mass spectrum of known cells. As an example, we applied this technique to verify subpopulations of human fibroblasts with different origins and which exhibit different medical characteristics. PMID- 19184976 TI - Cloning and immunoreactivity of the 5-HT 1Mac and 5-HT 2Mac receptors in the central nervous system of the freshwater prawn Macrobrachium rosenbergii. AB - Biogenic amines are implicated in several mental disorders, many of which involve social interactions. Simple model systems, such as crustaceans, are often more amenable than vertebrates for studying mechanisms underlying behaviors. Although various cellular responses of biogenic amines have been characterized in crustaceans, the mechanisms linking these molecules to behavior remain largely unknown. Observed effects of serotonin receptor agonists and antagonists in abdomen posture, escape responses, and fighting have led to the suggestion that biogenic amine receptors may play a role in modulating interactive behaviors. As a first step in understanding this potential role of such receptors, we have cloned and fully sequenced two serotonin receptors, 5-HT(1Mac) and 5-HT(2Mac), from the CNS of the freshwater prawn Macrobrachium rosenbergii and have mapped their CNS immunohistochemical distribution. 5-HT(1Mac) was found primarily on the membranes of subsets of cells in all CNS ganglia, in fibers that traverse all CNS regions, and in the cytoplasm of a small number of cells in the brain and circum- and subesophageal ganglia (SEG), most of which also appear to contain dopamine. The pattern of 5-HT(2Mac) immunoreactivity was found to differ significantly; it was found mostly in the central neuropil area of all ganglia, in glomeruli of the brain's olfactory lobes, and in the cytoplasm of a small number of neurons in the SEG, thoracic, and some abdominal ganglia. The observed differences in terms of localization, distribution within cells, and intensity of immunoreactive staining throughout the prawn's CNS suggest that these receptors are likely to play different roles. PMID- 19184979 TI - Cancer stem cells: a guide for skeptics. AB - Cancer stem cells (CSC) were postulated to exist many years ago as cells within a tumor that regenerate the tumor following treatment. A stochastic clonal evolution model was used to explain observed tumor heterogeneity. Recently, xenotransplantation studies have demonstrated that prospectively identifiable subpopulations from human cancers can initiate tumors in immune deficient mice, and these results along with recent advances in stem cell biology have generated much excitement in the cancer field. The modern CSC theory posits a hierarchy of cells analogous to normal stem cell development. Some controversy remains, however, as to whether these tumor initiating cells truly represent CSC, and whether the modern CSC field can live up to the promise of providing improved cancer treatments based on a novel model of cancer biology. Recent data from CSC investigators are discussed critically. PMID- 19184980 TI - The small GTPase RhoA is crucial for MC3T3-E1 osteoblastic cell survival. AB - Prolongation of cell survival through prevention of apoptosis is considered to be a significant factor leading to anabolic responses in bone. The current studies were carried out to determine the role of the small GTPase, RhoA, in osteoblast apoptosis, since RhoA has been found to be critical for cell survival in other tissues. We investigated the effects of inhibitors and activators of RhoA signaling on osteoblast apoptosis. In addition, we assessed the relationship of this pathway to parathyroid hormone (PTH) effects on apoptotic signaling and cell survival. RhoA is activated by geranylgeranylation, which promotes its membrane anchoring. In serum-starved MC3T3-E1 osteoblastic cells, inhibition of geranylgeranylation with geranylgeranyl transferase I inhibitors increased activity of caspase-3, a component step in the apoptosis cascade, and increased cell death. Dominant negative RhoA and Y27632, an inhibitor of the RhoA effector Rho kinase, also increased caspase-3 activity. A geranylgeranyl group donor, geranylgeraniol, antagonized the effect of the geranylgeranyl transferase I inhibitor GGTI-2166, but could not overcome the effect of the Rho kinase inhibitor. PTH 1-34, a potent anti-apoptotic agent, completely antagonized the stimulatory effects of GGTI-2166, dominant negative RhoA, and Y27632, on caspase 3 activity. The results suggest that RhoA signaling is essential for osteoblastic cell survival but that the survival effects of PTH 1-34 are independent of this pathway. PMID- 19184982 TI - Top-down proteomics with a quadrupole time-of-flight mass spectrometer and collision-induced dissociation. AB - With slight modifications of the instrumental parameters, we demonstrate that satisfactory top-down data can be obtained with collision-induced dissociation (CID) tandem mass spectrometry on a quadrupole time-of-flight (qTOF) instrument not originally designed for this purpose. Protein identification is achieved with both N- and C-terminal sequence tags and BLAST database searches. The accurate mass measurement of multiply charged fragment ions (mostly y and b-type) supplements the limited set of cleavage sites and provides a high degree of sequence coverage (90-100%). Post-translational modification issues can be addressed too. This approach might help those mass spectrometry (MS) core facilities that are not able to afford very high-resolution instruments, thus expanding the benefits of top-down protein analysis over the worldwide MS community. PMID- 19184981 TI - NMR assignments and histone specificity of the ING2 PHD finger. AB - The ING2 plant homeodomain (PHD) finger is recruited to the nucleosome through specific binding to histone H3 trimethylated at lysine 4 (H3K4me3). Here, we describe backbone and side chain assignments of the ING2 PHD finger, analyze its binding to the unmodified and modified histone and p53 peptides, and map the histone H3 and H3K4me3 binding sites based on chemical shift perturbation analysis. PMID- 19184983 TI - Abdominal wall reconstruction utilizing posterior rectus sheath fascia: a spare part for all reasons. PMID- 19184984 TI - Vitamin D receptor modulates the neoplastic phenotype through antagonistic growth regulatory signals. AB - Vitamin D receptor (VDR) can modulate functionally antagonistic growth regulatory pathways, involving beta-catenin/E-cadherin on one hand and osteopontin (OPN) on the other. This study investigates effects of VDR ligand treatment on the balance of these discordant signals and on associated cell behavior. Treatment of Rama 37 or SW480 cells by 1alpha,25-(OH)(2) D(3) or analogs suppressed beta-catenin/Lef 1/Tcf signaling and upregulated E-cadherin, consistent with a cancer-inhibitory action. Conversely, treatment also increased transcription of OPN that may be implicated in tumor progression. Molecular crosstalk was observed between the antagonistic VDR-dependent signals, in that beta-catenin/Lef-1/Tcf molecules modulated VDR activation of OPN. Treatment effects on cell growth were related to a constitutive balance of OPN and E-cadherin expression. No growth effects were observed in Rama 37 cells that have low OPN and high E-cadherin expression. Conversely, treatment of Rama 37 stably transfected subclones that had high OPN and/or low level E-cadherin induced small but significant increases of cell attachment to fibronectin, anchorage-independent growth or invasion. This study shows that relative expression levels of key VDR downstream genes may influence growth regulation by 1alpha,25-(OH)(2) D(3) or analogs. These findings may be relevant to the cell- or tissue-specificity of vitamin D growth regulation. PMID- 19184985 TI - Hepatocyte growth factor induces hypoxia-related interleukin-8 expression in lung adenocarcinoma cells. AB - Rapid growth of cancer cells often creates insufficient supply of oxygen and nutrients in the tumour nest. The frequent detection of hypoxia-inducible factor (HIF) and interleukin-8 (IL-8) in afflicted tissues suggests that IL-8 expression could be associated with elevated levels of HIF. Recently, we found that hypoxia also upregulated the expression of hepatocyte growth factor (HGF) in lung adenocarcinoma (LAD) cells. However, the relationship between HGF and IL-8 has not been investigated in LAD cells. In this study, we found that HGF induced IL-8 expression in LAD. Interestingly, hypoxia also increased the level of prostaglandin F(2alpha) (PGF(2alpha)), a product of dihydrodiol dehydrogenase (DDH). When expression of DDH was suppressed by siRNA, the levels of PGF(2alpha), HGF and IL-8 were reduced; however, their levels returned to normal after DDH was reintroduced. These data suggest that hypoxia induces biosynthesis of PGF(2alpha), which then activates HGF and IL-8 expression. The results provide a reasonable explanation of how PGF(2alpha), HGF and IL-8 exert their effects on cancer cell metastasis. PMID- 19184986 TI - Life success of males on nonoffender, adolescence-limited, persistent, and adult onset antisocial pathways: follow-up from age 8 to 42. AB - A random sample of 196 males, drawn from the Jyvaskyla Longitudinal Study of Personality and Social Development, was divided into four groups of offenders using information from government registers of convictions between ages 21 and 47, from local police registers searched at age 21, from a Self-Report Delinquency Scale administered at age 36, from a Life History Calendar for ages 15-42, and from personal interviews at ages 27, 36, and 42. The groups were: persistent offenders (offences before and after age 21; 29% of the men); adolescence-limited offenders (offences before age 21; 27%); adult-onset offenders (offences after age 21; 16%); and nonoffenders (28%). The profile of the persistent offenders included many characteristics of antisocial personality and problems of social and psychological functioning at all ages. In childhood and adolescence, the adolescence-limited offenders tended to display higher self control than the other offenders indicated by less negative emotionality and higher constraint. In early middle age, they were well established in terms of work and marriage, but they differed from the nonoffenders in higher neuroticism, aggressiveness, and psychosomatic symptoms as did the other offender groups. The adult-onset offenders displayed an emotionally unstable personality, and in adulthood, they tended to be heavy users of alcohol. Parental SES did not differentiate the groups, but the parents of the persistent offenders tended to use alcohol heavily, and the social capital of the family tended to be low compared with the other groups. PMID- 19184987 TI - Crystalline nanorods as possible templates for the synthesis of amorphous biosilica during spicule formation in Demospongiae. AB - High-resolution microscopy shows that, during the initial stages of demosponge spicule formation, a primordial crystalline structure is formed within the axial filament. The recently developed electron diffraction tomography technique reveals that the nanorods have a layered structure that matches smectitic phyllosilicates. These intracellular nanorods have been considered as precursors of mature spicules. PMID- 19184988 TI - Fluorescence detection of a protein-bound 2Fe2S cluster. AB - A fluorescent biosensor is described for 2Fe2S clusters that is composed of green fluorescent protein (GFP) fused to glutaredoxin 2 (Grx2), as illustrated here. 2Fe2S detection is based on the reduction of GFP fluorescence upon the 2Fe2S induced dimerization of GFP-Grx2. This assay is sufficiently sensitive to detect submicromolar changes in 2Fe2S levels, thus making it suitable for high throughput measurements of metallocluster degradation and synthesis reactions. PMID- 19184989 TI - A peptide antagonist of the TLR4-MD2 interaction. AB - Toll-like receptors are an integral part of innate immunity in the central nervous system (CNS); they orchestrate a robust defense in response to both exogenous and endogenous danger signals. Recently, toll-like receptor 4 (TLR4) has emerged as a therapeutic target for the treatment of CNS-related diseases such as sepsis and chronic pain. We herein report a chemical biology approach by using a rationally designed peptide inhibitor to disrupt the TLR4-MD2 association, thereby blocking TLR4 signaling. PMID- 19184990 TI - Multiplexed labeling of viable cells for high-throughput analysis of glycine receptor function using flow cytometry. AB - Flow cytometry is an important drug discovery tool because it permits high content multiparameter analysis of individual cells. A new method dramatically enhanced screening throughput by multiplexing many discrete fixed cell populations; however, this method is not suited to assays requiring functional cellular responses. HEK293 cells were transfected with unique mutant glycine receptors. Mutant receptor expression was confirmed by coexpression of yellow fluorescent protein (YFP). Commercially available cell-permeant dyes were used to label each glycine receptor expressing mutant with a unique optical code. All encoded cell lines were combined in a single tube and analyzed on a flow cytometer simultaneously before and after the addition of glycine receptor agonist. We decoded multiplexed cells that expressed functionally distinct glycine receptor chloride channels and analyzed responses to glycine in terms of chloride-sensitive YFP expression. Here, data provided by flow cytometry can be used to discriminate between functional and nonfunctional mutations in the glycine receptor, a process accelerated by the use of multiplexing. Further, this data correlates to data generated using a microscopy-based technique. The present study demonstrates multiplexed labeling of live cells, to enable cell populations to be subject to further cell culture and experimentation, and compares the results with those obtained using live cell microscopy. PMID- 19184991 TI - Safety and efficacy of sonographic-guided random real-time core needle biopsy of the liver. AB - PURPOSE: To determine the safety and efficacy of real-time, sonographic-guided, random percutaneous needle biopsy of the liver in a tertiary medical center. METHOD: From an IRB-approved biopsy database, all patients who had random liver biopsy performed over a 24-month period were selected. In 350 patients, 539 random percutaneous needle biopsies of the liver were performed under real-time sonographic visualization. The following were recorded from the electronic medical record: patient demographics, indication for biopsy procedure; radiologist's name; needle type and gauge and number of passes; use and amount of i.v. sedation or anesthesia; adequacy of the specimen; and complications following the procedure. RESULT: Of 539 biopsies, 378 (70%) biopsy procedures were performed on liver transplant recipients. Of the biopsy procedures in nontransplant patients, 81/161 (50%) concurrently underwent biopsy of a focal liver mass. An 18-gauge automated core biopsy needle was used in 536/539 (99%). Median number of passes per biopsy procedure was 1 (mean, 1.7; range, 1-6). Sedation using midazolam and fentanyl was used in 483/539 (90%). There were only 8 inadequate specimens (1.5%, [2.3, upper 95% confidence limit, fully described in Statistical Analysis]). Complications were identified in 11/539 biopsy procedures (2.0%, [2.6, upper 95% confidence limit]): 5 with severe postprocedural pain, 3 with symptomatic hemorrhage, 2 with infection, and 1 with a rash. There were no sedation-related complications and no deaths related to the procedure. CONCLUSION: Real-time, sonographic-guided, random core-needle liver biopsy is a safe and highly effective procedure. PMID- 19184992 TI - Sonography of the pediatric thyroid: a pictorial essay. AB - The purpose of this article is to review the various pediatric thyroid diseases using sonographic images. These diseases included congenital hypothyroidism (dysgenesis [aplasia; ectopy; hypoplasia], dyshormonogenesis, transient hypothyroidism), thyroglossal duct cyst, Graves' disease, Hashimoto's thyroiditis, suppurative thyroiditis, nodular hyperplasia (goiter) and thyroid mass. Thyroid sonogram is the primary method used for thyroid disease and can be used for guiding of fine needle aspiration to obtain a more accurate diagnosis for suspected thyroid lesions. PMID- 19184993 TI - Off-line sentence processing: what is involved in answering a comprehension probe? AB - The aim of this study was to better characterize the influence of the comprehension probe on syntax-related activation patterns observed in fMRI studies of sentence comprehension. In this study, sentence comprehension was assessed by presenting a true/false statement after each sentence. To disassociate the sentence reading from the comprehension probe activation, a 6-s delay was placed between these processing phases. Two factors were manipulated, one affected the sentence and the other affected the probe. The sentences were manipulated by varying their syntactic complexity; conjoined-active and object relative sentences were examined. The comprehension probes asked whether one of the first two mentioned nouns in the preceding sentence performed the action of one of the two verbs. The probes were manipulated by varying the distance (number of intervening words) between the noun and verb within the sentence; there were three probe types: short distance, long distance, and false statements. The results, which focused on the processing taking place during the probe, showed that the distance manipulation resulted in significant differences in both behavioral and brain activation measures. This was particularly true of BA 44, which revealed an interaction between complexity and distance such that the complexity effect was all but eliminated for the long-distance condition. Additionally, we replicated our previous finding of syntactic complexity effects during the probe phase. Finally, post hoc analysis revealed that participants used two distinct strategies during sentence reading; significant effects of strategy use on both behavioral and brain activation data were observed. PMID- 19184994 TI - Random fields--union intersection tests for detecting functional connectivity in EEG/MEG imaging. AB - Electrophysiological (EEG/MEG) imaging challenges statistics by providing two views of the same underlying spatio-temporal brain activity: a topographic view (EEG/MEG) and tomographic view (EEG/MEG source reconstructions). It is a common practice that statistical parametric mapping (SPM) for these two situations is developed separately. In particular, assessing statistical significance of functional connectivity is a major challenge in these types of studies. This work introduces statistical tests for assessing simultaneously the significance of spatio-temporal correlation structure between ERP/ERF components as well as that of their generating sources. We introduce a greatest root statistic as the multivariate test statistic for detecting functional connectivity between two sets of EEG/MEG measurements at a given time instant. We use some new results in random field theory to solve the multiple comparisons problem resulting from the correlated test statistics at each time instant. In general, our approach using the union-intersection (UI) principle provides a framework for hypothesis testing about any linear combination of sensor data, which allows the analysis of the correlation structure of both topographic and tomographic views. The performance of the proposed method is illustrated with real ERP data obtained from a face recognition experiment. PMID- 19184995 TI - Strength of prefrontal activation predicts intensity of suggestion-induced pain. AB - Suggestion, a powerful factor in everyday social interaction, is most effective during hypnosis. Subjective evaluations and brain-imaging findings converge to propose that hypnotic suggestion strongly modulates sensory processing. To reveal the brain regions that mediate such a modulation, we analyzed data from a functional-magnetic-resonance-imaging study on hypnotic-suggestion-induced pain on 14 suggestible subjects. Activation strengths in the right dorsolateral prefrontal cortex (DLPFC) during initiation of suggestion for pain correlated positively with the subjective intensity of the subsequent suggestion-induced pain, as well as with the strengths of the maximum pain-related activation in the in the secondary somatosensory (SII) cortex. Furthermore, activation of the insula and the anterior cingulate cortex predicted the pain-related SII activation. The right DLPFC, as an area important for executive functions, likely contributes to functional modulation in the modality-specific target areas of given suggestions. PMID- 19184996 TI - Cell-cell-neighborhood relations in tissue sections--a quantitative model for tissue cytometry. AB - Physical interactions between different cell types are a requirement for the initiation and maintenance of immune responses. The distribution pattern of cells within a tissue may result from specific cell-cell-interactions or random distribution. Tissue architecture, degree of inflammation, frequencies of cells, number of contact partners, cell type, and size as well as cell movement and contact time determine the distribution of cells within tissues. We developed a matrix model to determine the degree of expected random distribution of two cell types (A and B) and cell-cell-contacts within tissue sections. The model predictions were compared with experimental data derived from immunofluorescence microscopy. We implemented a computer algorithm for automatic image analysis to visualize and quantify cell-cell-neighborhood relations. Using the number of cells type A (a), the total cell number (t) and the mean number of cells that are in contact with cells type B (c(B)), the ratio of cells type B in contact with cells type A can be described by b(A)/b = 1- (1- (a/t))[symbol: see text]c(B). We applied the model system to investigate the distribution of Foxp3(+) regulatory T cells with Ki-67(+) proliferating cells within mouse tissue sections. The matrix model provides a tool to describe the expected distribution of two different cell types and their cell-cell-contacts within tissues. Comparing the degree of expected random distribution with experimental data might help to propose functional cell-cell-interactions in tissue sections. PMID- 19184997 TI - Dopaminergic modulation of brain systems subserving decision making under uncertainty: a study with fMRI and methylphenidate challenge. AB - There is evidence that the dopaminergic system is involved in probabilistic reinforcement learning and reward-related decision-making. However, little is known about the effects of external dopaminergic challenges on processing of uncertainty in decision-making tasks. Therefore, the present study examined changes in fMRI activation patterns in a natural sampling paradigm. Decision making under uncertainty was examined before and after administration of a single dose of 40 mg methylphenidate as an acute dopaminergic pharmacological challenge. We found that the level of uncertainty was positively correlated with activations in the prefrontal cortex. Conversely, negative correlations with uncertainty were found in the left hippocampus, right amygdale, and right middle temporal gyrus. The drug intervention with methylphenidate revealed a differential picture. Uncertain information processing was associated with higher activation in the parietal association cortex and posterior cingulate cortex after placebo relative to methylphenidate. The methylphenidate challenge relative to placebo was associated with higher left and right parahippocampal as well as cerebellar activation under uncertainty. Apparently, the pro-dopaminergic pharmacological influence induces a relative shift towards recruitment of hippocampal areas under uncertainty, whereas under placebo conditions, higher levels of parietal cortex activations are involved in the task. The findings suggest a role of dopamine in uncertainty processing and shed light on the pharmacological mechanisms of methylphenidate. PMID- 19184998 TI - Rats with persistently high exploratory activity have both higher extracellular dopamine levels and higher proportion of D(2) (High) receptors in the striatum. AB - Increases in both striatal dopamine release and the proportion of the D(2) receptors in the high affinity state (D(2) (High)) accompany the behavioral sensitization to psychostimulants, but it is not known whether the physiological substrate of the interindividual differences locomotor and exploratory behavior is similar. Thus, we examined whether persistently high spontaneous exploratory activity is associated with extracellular dopamine as well as the proportion of D(2) (High) in the striatum. Extracellular dopamine levels were found to be significantly higher in rats with high exploratory activity (high explorers, HE) as compared with low explorers (LE) in baseline conditions as well as after administration of amphetamine (0.5 mg/kg, i.p.). Also, the HE animals had significantly higher proportion of striatal D(2) (High) receptors than the LE rats (43.8 +/- 4.4% and 22.5 +/- 1.5%, respectively). Thus, the present findings support the notion that concomitant higher extracellular dopamine levels and the proportion of D(2) (High) receptors in the striatum, whether naturally occurring and persistent or pharmacologically induced, are causally related to high behavioral activity. PMID- 19184999 TI - Diagnosis of recurrent lung cancer in the mediastinum using endosonographically guided fine-needle aspiration biopsy. AB - Endoscopic sonography is a well-established tool for the diagnosis and staging of gastrointestinal malignancies. It has been found to be of increasing utility in the diagnosis and management of lung cancers. We report a case of recurrent lung cancer presenting 4 years after initial diagnosis and treatment, in which endoscopic sonography was used to guide the fine-needle aspiration biopsy. PMID- 19185000 TI - Apotopes and the biliary specificity of primary biliary cirrhosis. AB - Primary biliary cirrhosis (PBC) is characterized by antimitochondrial antibodies (AMAs), directed to the E2 component of the pyruvate dehydrogenase complex (PDC E2). Notwithstanding the presence of mitochondria in virtually all nucleated cells, the destruction in PBC is limited to small intrahepatic bile ducts. The reasons for this tissue specificity remain unknown, although biliary epithelial cells (BECs) uniquely preserve the PDC-E2 epitope following apoptosis. Notably, PBC recurs in an allogeneic transplanted liver, suggesting generic rather than host PBC-specific susceptibility of BEC. We used cultured human intrahepatic BECs (HIBECs) and other well-characterized cell lines, including, HeLa, CaCo-2 cells, and nontransformed human keratinocytes and bronchial epithelial cells, to determine the integrity and specific localization of PDC-E2 during induced apoptosis. All cell lines, both before and after apoptosis, were tested with sera from patients with PBC (n = 30), other autoimmune liver and rheumatic diseases (n = 20), and healthy individuals (n = 20) as well as with a mouse monoclonal antibody against PDC-E2 and AMA with an immunoglobulin A isotype. PDC-E2 was found to localize unmodified within apoptotic blebs of HIBECs, but not within blebs of various other cell lineages studied. The fact that AMA-containing sera reacted with PDC-E2 on apoptotic BECs without a requirement for permeabilization suggests that the autoantigen is accessible to the immune system during apoptosis. CONCLUSION: Our data indicate that the tissue (cholangiocyte) specificity of the autoimmune injury in PBC is a consequence of the unique characteristics of HIBECs during apoptosis and can be explained by exposure to the immune system of intact immunoreactive PDC-E2 within apoptotic blebs. PMID- 19185001 TI - Gene expression signatures of peripheral CD4+ T cells clearly discriminate between patients with acute and chronic hepatitis B infection. AB - CD4+ T and regulatory T cells (Tregs) seem to play a key role in persistence of hepatitis B virus (HBV) infection. However, the molecular events by which Tregs exert their modulatory activity are largely unknown. The transcriptional profiles of CD4+ T cells of healthy controls (HCs) and patients affected by acute hepatitis B (AVH-B) or chronic hepatitis B (CHB) infection were established using a custom expression array consisting of 350 genes relevant for CD4+ T cell and Treg function. These studies were complemented by real-time reverse-transcription polymerase chain reaction. Peripheral blood mononuclear cells (PBMCs) were also analyzed for the presence of Tregs, which were more abundant in the acute stage of the disease (7%) than in HCs and CHB infection (HCs versus AVH-B, P = 0.003; AVH-B versus CHB, P = 0.04). One hundred eighteen genes (34%) intrinsically differentiate HBV-infected patients from HCs. Using gene ontology, we identified T cell receptor signaling and clusterization, mitogen-activated protein kinase kinase signaling, cell adhesion, cytokines and inflammatory responses, cell cycle/cell proliferation, and apoptosis as the most prominent affected modules. A higher expression of CCR1, CCR3, CCR4, CCR5, and CCR8 was seen in AVH-B than in CHB-infected patients and HCs. Annotation of the interconnected functional network of genes provided a unique representation of global immune activation during acute infection. Almost all genes were down-regulated in patients with CHB infection. CONCLUSION: The fingerprints enable clear discrimination between patients suffering from AVH-B or CHB infection. The observed profiles suggest accumulation of effector T cells with a potential role in necro-inflammation during the acute stage. Subsequent down-regulated effector functions support the hypothesis of suppressed CD4+ effector T cells favoring viral persistence in the chronic infection stage. PMID- 19185002 TI - Reproducibility assessment of a multiple reference tissue method for quantitative dynamic contrast enhanced-MRI analysis. AB - Bone metastases of 16 prostate cancer patients were scanned twice 1 week apart by dynamic contrast enhanced (DCE)-MRI at 2-s resolution using a two-dimensional gradient-echo pulse sequence. With a multiple reference tissue method (MRTM), the local tissue arterial input function (AIF) was estimated using the contrast agent enhancement data from tumor subregions and muscle. The 32 individual AIFs estimated by the MRTM, which had considerable intra-patient and inter-patient variability, were similar to directly measured AIFs in the literature and using the MRTM AIFs in a pharmacokinetic model to derive estimated individual cardiac outputs provided physiologically reasonable results. The MRTM individual AIFs gave better fits with smaller sum of squared errors and equally reproducible estimate of kinetic parameters compared with a previous reported population AIF measured from remote arteries. The individual MRTM AIFs were also used to obtain a mean local tissue AIF for the unique population of this study, which further improved the reproducibility of the estimated kinetic parameters. The MRTM can be applied to DCE-MRI studies of bone metastases from prostate cancers to provide an AIF from which reproducible quantitative DCE-MRI parameters can be derived, thus help standardize DCE-MRI studies in cancer patients. PMID- 19185003 TI - A nonessential role for Arg 55 in cyclophilin18 for catalysis of proline isomerization during protein folding. AB - The protein folding process is often in vitro rate-limited by slow cis-trans proline isomerization steps. Importantly, the rate of this process in vivo is accelerated by prolyl isomerases (PPIases). The archetypal PPIase is the human cyclophilin 18 (Cyp18 or CypA), and Arg 55 has been demonstrated to play a crucial role when studying short peptide substrates in the catalytic action of Cyp18 by stabilizing the transition state of isomerization. However, in this study we show that a R55A mutant of Cyp18 is as efficient as the wild type to accelerate the refolding reaction of human carbonic anhydrase II (HCA II). Thus, it is evident that the active-site located Arg 55 is not required for catalysis of the rate-limiting prolyl cis-trans isomerization steps during the folding of a protein substrate as HCA II. Nevertheless, catalysis of cis-trans proline isomerization in HCA II occurs in the active-site of Cyp18, since binding of the inhibitor cyclosporin A abolishes rate acceleration of the refolding reaction. Obviously, the catalytic mechanisms of Cyp18 can differ when acting upon a simple model peptide, four residues long, with easily accessible Pro residues compared with a large protein molecule undergoing folding with partly or completely buried Pro residues. In the latter case, the isomerization kinetics are significantly slower and simpler mechanistic factors such as desolvation and/or strain might operate during folding-assisted catalysis, since binding to the hydrophobic active site is still a prerequisite for catalysis. PMID- 19185004 TI - A missense mutation in ABCB4 gene involved in progressive familial intrahepatic cholestasis type 3 leads to a folding defect that can be rescued by low temperature. AB - Progressive familial intrahepatic cholestasis type 3 (PFIC3) is a rare liver disease characterized by early onset of cholestasis that leads to cirrhosis and liver failure before adulthood. PFIC3 may be improved by chronic administration of ursodeoxycholic acid, although in many cases liver transplantation is the only therapy. The disease is caused by mutations of the adenosine triphosphate (ATP) binding cassette, sub-family B, member 4 (ABCB4) [multidrug resistance 3 (MDR3)] gene encoding a specific hepatocellular canalicular transporter involved in biliary phosphatidylcholine secretion. Several mutations have been reported; however, the effect of individual mutations has not been investigated. ABCB4 is highly homologous to ATP-binding cassette, sub-family B, member 1 (ABCB1) (MDR1), the multidrug transporter responsible for drug resistance of cancer cells. We have studied the effect of mutation I541F localized to the first nucleotide binding domain, which is highly conserved between ABCB4 and ABCB1. Plasmids encoding the wild-type human ABCB4 or rat ABCB1-green fluorescing protein (GFP) construct, and corresponding I541F-mutants, were expressed in hepatocellular carcinoma, human (HepG2) and Madin-Darby canine kidney (MDCK) cells. Expression studies showed that ABCB4 was localized at the bile canalicular membrane in HepG2 cells and at the apical surface in MDCK cells, whereas the I541F mutant was intracellular. In MDCK cells, ABCB1-I541F also accumulated intracellularly in compartments, which were identified as the endoplasmic reticulum and cis-Golgi, and remained partially endoH-sensitive. After shifting cells to 27 degrees C, ABCB1-I541F was expressed at the apical cell surface in a mature and active form. Similarly, ABCB4 was significantly trafficked to the membrane of bile canaliculi in HepG2 cells. CONCLUSION: Mutation I541F causes mislocalization of both ABCB4 and ABCB1. Intracellular retention of ABCB4-I541F can explain the disease in PFIC3 patients bearing this mutation. The observation that plasma membrane expression and activity can be rescued by low temperature opens perspectives to develop novel therapies for the treatment of PFIC3. PMID- 19185005 TI - High expression of the bile salt-homeostatic hormone fibroblast growth factor 19 in the liver of patients with extrahepatic cholestasis. AB - Fibroblast growth factor 19 (FGF19) is an endocrine factor produced by the small intestine in response to uptake of luminal bile salts. In the liver, FGF19 binds to FGF receptor-4, resulting in down-regulation of cytochrome P (CYP) 7A1 and reduced bile salt synthesis. Down-regulation of CYP7A1 under cholestatic conditions has been attributed to bile salt-mediated induction of the transcriptional repressor short heterodimer partner (SHP), because the interrupted enterohepatic cycle of bile salts is thought to abrogate intestinal FGF19 production and thus result in lowering of plasma FGF19 levels. Unexpectedly, we observed marked elevation of plasma FGF19 in patients with extrahepatic cholestasis caused by a pancreatic tumor (2.3 +/- 2.3 in cholestatic versus 0.40 +/- 0.25 ng/mL and 0.29 +/- 0.12 ng/mL in postcholestatic patients who received preoperative drainage by biliary stenting, P = 0.004, and noncholestatic control patients, P = 0.04, respectively). Although FGF19 messenger RNA (mRNA) is virtually absent in normal liver, FGF19 mRNA was strongly increased (31-fold to 374-fold, P < 0.001) in the liver of cholestatic patients in comparison with drained and control patients. In the absence of changes in SHP mRNA, CYP7A1 mRNA was strongly reduced (7.2-fold to 24-fold, P < 0.005) in the liver of cholestatic patients in comparison with drained and control patients, indicating an alternative regulatory pathway. Alterations in transcripts encoding hepatobiliary transporters [adenosine triphosphate-binding cassette, subfamily C, member 3 (ABCC3)/multidrug resistance protein 3 (MRP3), organic solute transporter alpha/beta (OSTalpha/beta), organic anion-transporting polypeptide (OATP1B1)] further suggest that bile salts are secreted via a nonbiliary route in patients with extrahepatic cholestasis. CONCLUSION: The liver expresses FGF19 under conditions of extrahepatic cholestasis. This is accompanied by a number of adaptations aimed at protecting the liver against bile salt toxicity. FGF19 signaling may be involved in some of these adaptations. PMID- 19185006 TI - Bile salt-induced pro-oxidant liver damage promotes transplanted cell proliferation for correcting Wilson disease in the Long-Evans Cinnamon rat model. AB - Insights into disease-specific mechanisms for liver repopulation are needed for cell therapy. To understand the efficacy of pro-oxidant hepatic perturbations in Wilson disease, we studied Long-Evans Cinnamon (LEC) rats with copper toxicosis under several conditions. Hepatocytes from healthy Long-Evans Agouti (LEA) rats were transplanted intrasplenically into the liver. A cure was defined as lowering of copper to below 250 microg/g liver, presence of ATPase, Cu++ transporting, beta polypeptide (atp7b) messenger RNA (mRNA) in the liver and improvement in liver histology. Treatment of animals with the hydrophobic bile salt, cholic acid, or liver radiation before cell transplantation produced cure rates of 14% and 33%, respectively; whereas liver radiation plus partial hepatectomy followed by cell transplantation proved more effective, with cure in 55%, P < 0.01; and liver radiation plus cholic acid followed by cell transplantation was most effective, with cure in 75%, P < 0.001. As a group, cell therapy cures in rats preconditioned with liver radiation plus cholic acid resulted in less hepatic copper, indicating greater extent of liver repopulation. We observed increased hepatic catalase and superoxide dismutase activities in LEC rats, suggesting chronic oxidative stress. After liver radiation or cholic acid, hepatic lipid peroxidation levels increased, indicating further oxidative injury, although we did not observe overt additional cytotoxicity. This contrasted with healthy animals in which liver radiation and cholic acid produced hepatic steatosis and loss of injured hepatocytes. We concluded that pro-oxidant perturbations were uniquely effective for cell therapy in Wilson disease because of the nature of preexisting hepatic damage. PMID- 19185007 TI - Accuracy of Thinprep Imaging System in detecting atypical glandular cells. AB - The aim of this study was to evaluate the accuracy of the ThinPrep Imaging System (Imager) in detecting atypical glandular cells (AGC) or adenocarcinoma in the 22 selected fields. All cases reported as AGC or adenocarcinoma from January 2005 to December 2006 that had been initially screened by the Imager were retrospectively reviewed to determine whether the most diagnostically relevant groups/cells were within the 22 selected fields. A total of 39 cases were reviewed. The cases were divided into two groups: the group with diagnostic cells detected within the 22 selected fields (accurately detected group) and the group with upgraded diagnosis following rescreening process (underdetected group). The Imager accurately detected 32 (82%) of cases with abnormal glandular cells, including six cases reported as adenocarcinoma, one case as adenocarcinoma in situ (AIS), and 25 cases as AGC. In seven (18%) cases, the Imager failed to detect the most abnormal cells within the 22 selected fields. Among these, one case was adenocarcinoma, while the rest were reported as AGC. Overall, four cases were assessed as atypical during quality control (QC) rescreening even though the Imager detected abnormal groups in most. Fourteen of 32 cases had abnormalities proven by histologic follow-up. Overall, the Imager was effective in detecting most AGC, AIS and invasive adenocarcinomas. In a minority (18%) of cases, the Imager failed to detect the cells of interest within the 22 selected fields. However, full manual review of cases with potential atypical/reactive groups or endometrial groups in women > or = 40 and QC rescreening of selected cases may help to minimize the underdetected cases. PMID- 19185008 TI - (123)I-mIBG scintigraphy in patients with known or suspected neuroblastoma: Results from a prospective multicenter trial. AB - BACKGROUND: A prospective trial was conducted to confirm the diagnostic performance of (123)I-mIBG scintigraphy in patients with known or suspected neuroblastoma. PROCEDURE: One hundred patients (mean age 4.7 years) were enrolled, 86 with a previous diagnosis of neuroblastoma, 13 with suspected disease based upon symptoms, imaging findings, and elevated catecholamines, and one adult with an abdominal tumor thought to be of neuroendocrine origin. All patients underwent whole-body planar imaging 24 hr following IV administration of 1-10 mCi (37-370 MBq) (123)I-mIBG. SPECT imaging of the thorax/abdomen/pelvis was performed in 51 patients. Images were interpreted by three blinded readers, with consensus requiring agreement of at least two readers. Disease status was confirmed by histopathology, imaging results, catecholamine measurements, and follow-up. RESULTS: Sixty-four patients had active disease, 30 were without disease, and 6 were judged indeterminate because of insufficient confirmatory data. (123)I-mIBG scintigraphy had a sensitivity of 88% (56/64) and specificity of 83% (25/30). Sensitivity was 91% (30/33) among the subset of subjects who had both planar and SPECT imaging. Among 53 patients with recent histopathology, sensitivity and specificity were 93% and 92%, respectively. Most false-negative interpretations were in patients with minimal residual disease (n = 4), while false-positive interpretations generally involved atypical adrenal or other physiological uptake (n = 4). CONCLUSIONS: This prospective multicenter trial of (123)I-mIBG scintigraphy documents high sensitivity and specificity of this imaging technique in patients with both newly diagnosed and previously treated neuroblastoma. PMID- 19185009 TI - Hits and misses in targeting pediatric cancers. PMID- 19185012 TI - Tramadol hydrochloride use and acute deterioration in Parkinson's disease tremor. PMID- 19185011 TI - How cautious should we be when assessing apathy with the Unified Parkinson's Disease Rating Scale? AB - Current practice often assesses apathy with a single item from the Unified Parkinson's Disease Rating Scale (UPDRS, item 4). Yet, the relationship between the UPDRS item 4 and the validated Apathy Scale (AS) is unknown. The purpose of this study was to evaluate the operating characteristics of UPDRS item 4 in relation to the AS. Three hundred and one patients with PD were administered the AS and the UPDRS. We compared the UPDRS item 4 to the standard AS classification of > or =14 as apathetic. A receiver operating characteristics (ROC) curve was obtained, and sensitivity, specificity, positive, and negative predictive power were calculated. The ROC curve showed area under the curve as 0.75. A cut-off of 1 had good sensitivity (81%) but poor specificity (53%; high false positive rate). A cut-off point of 2 had acceptable specificity (87%) but poor sensitivity (52%, high false negative rate). Continuing to increasing the cut-off point (e.g., 3, 4) continues to increase specificity at the expense of dramatically reducing sensitivity. These findings suggest the use of caution when screening for apathy with item 4 due to its poor sensitivity in relation to the AS. PMID- 19185013 TI - Serum cholesterol levels and the risk of multiple system atrophy: a case-control study. AB - Cholesterol in brain membranes may modulate the conformational state and accumulation of alpha-synuclein in alpha-synucleinopathies.We examined the association between serum cholesterol and the risk of multiple system atrophy (MSA), one of the alpha-synucleinopathies. We enrolled 142 patients with probable MSA from two tertiary referral hospitals and 155 age- and gender-matched healthy people with no neurological disease. The levels of total cholesterol, low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) were significantly lower in MSA patients than in controls (total cholesterol: 172.7 vs. 196.3 mg/dL, P < 0.001; LDL-C: 104.0 vs. 115.3 mg/dL, P = 0.001; HDL-C: 47.3 vs. 54.2 mg/dL, P < 0.001). After adjusting for age, gender, use of cholesterol-lowering drugs, and histories of hypertension, diabetes mellitus, and smoking, the odds ratios was 5.9 (95% CI = 2.3-11.5, P < 0.001) for MSA patients in the lowest quartile of total cholesterol and 2.6 (95% CI = 1.2-5.5, P = 0.016) for those in the lowest quartile of HDL-C, compared with the highest quartiles. Levels of serum cholesterol did not significantly correlate with disease duration or severity. Our data suggest that lower levels of total cholesterol and HDL may be associated with an increased risk of MSA. PMID- 19185014 TI - Complicated recessive dystonia parkinsonism syndromes. AB - In addition to pure PD and pure dystonic syndromes, there are a group of disorders with overlapping features. The differential diagnosis of these dystonia parkinsonism syndromes can be complex. In view of the growing list of recognized disorders and recent advances in genetics, we review the autosomal recessive forms of dystonia parkinsonism, summarizing clinical presentations, results of investigations, and response to treatment of gene-proven cases. We concentrate on PANK2-, PLA2G6-, ATP13A2-, FBX07, TAF1-, and PRKRA-associated neurodegeneration. Parkin, PINK1, and DJ-1 are also briefly reviewed. PMID- 19185015 TI - Comprehensive analysis of expression, subcellular localization, and cognate pairing of SNARE proteins in oligodendrocytes. AB - Oligodendrocytes form the central nervous system myelin sheath by spiral wrapping of their plasma membrane around axons, necessitating a high rate of exocytic membrane addition to the growing myelin membrane. Membrane fusion is mediated by soluble N-ethylmaleimide-sensitive factor attachment protein receptor proteins (SNAREs), which act by specific pairing of vesicle (R)- and target (Q)-SNAREs. To characterize oligodendroglial SNAREs and their trafficking pathways, we performed a detailed expression analysis of SNAREs in differentiating cultured oligodendrocytes and myelin and determined their subcellular localization. Expression of the plasma membrane Q-SNAREs syntaxin 3, syntaxin 4, SNAP23, and the endosomal R-SNARE VAMP3/cellubrevin increased with oligodendroglial maturation, while the expression of SNAP29 decreased. Interestingly, syntaxin 3, syntaxin 4, and VAMP7/tetanustoxin-insensitive VAMP accumulated in myelin during development, suggesting a role in myelin membrane fusion. Coimmunoprecipitation from oligodendroglial cell lysates elucidated interactions between SNAREs: for example, Golgi-localized VAMP4 associated with syntaxin 6 and SNAP29. Furthermore, we identified a cognate core complex composed of VAMP3, syntaxin 4, and SNAP23, which may mediate fusion of endosome-derived vesicles with the plasma membrane. This study provides a comprehensive analysis of SNARE proteins in oligodendrocytes and assigns defined SNAREs to putative vesicle trafficking pathways in myelinating oligodendrocytes, thus facilitating future functional analysis of distinct SNAREs in oligodendroglial membrane traffic and myelination. PMID- 19185016 TI - Intention tremor in essential tremor: Prevalence and association with disease duration. PMID- 19185017 TI - Increase in the tactile catchment area of a sensory trick for alleviating blepharospasm following pallidal DBS. PMID- 19185018 TI - Pseudohypoparathyroidism manifesting with paroxysmal dyskinesias and seizures. PMID- 19185020 TI - Hiccups associated with levodopa in Parkinson's disease. PMID- 19185021 TI - Effects of DBS, premotor rTMS, and levodopa on motor function and silent period in advanced Parkinson's disease. AB - Deep brain stimulation (DBS) of the subthalamic nucleus (STN) is a widely used and highly effective treatment for patients with advanced Parkinson's disease (PD). Repetitive TMS (rTMS) applied to motor cortical areas has also been shown to improve symptoms in PD and modulate motor cortical excitability. Here, we compared clinical and neurophysiological effects of STN stimulation with those of 1 Hz rTMS given to the dorsal premotor cortex (PMd) and those following intake of levodopa in a group of PD patients with advanced disease. Ten PD patients were studied on 2 consecutive days before and after surgery. Clinical effects were determined using the UPDRS motor score. Motor thresholds, motor-evoked potential (MEP) amplitudes during slight voluntary contraction, and the cortical silent periods (SP) were measured using TMS. Before surgery effects of levodopa and 1 Hz PMd rTMS and after surgery those of STN stimulation with or without additional levodopa were determined. Levodopa significantly improved clinical symptoms and increased the SP duration. STN stimulation improved clinical symptoms without changing the SP duration. In contrast, 1 Hz PMd rTMS was not effective clinically but normalized the SP duration. Whereas levodopa had widespread effects at different levels of an abnormally active motor network in PD, STN stimulation and PMd rTMS led to either clinical improvement or SP normalization, i.e., only partially reversed abnormal motor network activity. PMID- 19185019 TI - The influence of NF-kappaB signal-transduction pathways on the murine inner ear by acoustic overstimulation. AB - Nuclear factor-kappa B (NF-kappaB) comprises a family of inducible transcription factors that serve as important regulators of the host immune and inflammatory responses. The NF-kappaB signals are activated via the canonical and/or noncanonical pathways in response to diverse stimuli. The excessive action of NF kappaB signal-transduction pathways frequently causes self-injurious phenomena such as allergic diseases, vascular disorders, and ischemia-reperfusion neuronal damage. In the inner ear, the role of NF-kappaB has not been clarified because the activated NF-kappaB signals potentially induce both cytoprotective and cytotoxic target genes after ototoxic stimulation. In the present study, we investigated the response of NF-kappaB in both the canonical and noncanonical pathways to acoustic overstimulation (117 dB/SPL/2 hr) and followed the change of inflammatory factors (inducible nitric oxide synthase [iNOS], intracellular adhesion molecule-1 [ICAM-1], and vascular cell adhesion molecule-1 [VCAM-1]) in the cochlear lateral wall (CLW) and the rest of cochlea (RoC). By means of immunohistochemistry combined with confocal microscopy and reverse transcriptase polymerase chain reaction techniques, we found the response of NF-kappaB family members (NF-kappa B1, 2, RelA, and RelB) at the transcription level. After the NF kappaB signaling, the inflammatory factors were significantly increased in the CLW and the RoC. Additionally, at the protein level, the prominent expression of adhesion molecules (ICAM-1 and VCAM-1) was observed in the tissue around the capillaries in the stria vascularis. These results show that acoustic overstimulation causes the NF-kappaB signaling to overexpress the inflammatory factors in the inner ear, and the up-regulation of the adhesion molecules (ICAM-1 and VCAM-1) and iNOS potentially influence the hemodynamics and the cellular integrity in the stria vascularis. PMID- 19185022 TI - Ginsenosides protect striatal neurons in a cellular model of Huntington's disease. AB - Ginseng, the root of Panax ginseng C.A. Meyer (Araliaceae), is a widely used herbal medicine. Ginsenosides, the active ingredients of ginseng, are the main components responsible for many beneficial actions of ginseng. In the present study, we tested 10 different ginsenosides in the previously developed in vitro Huntington's disease (HD) assay with primary medium spiny striatal neuronal cultures (MSN) from the YAC128 HD mouse model. We found that nanomolar concentrations of ginsenoside Rb1 and Rc effectively protected YAC128 medium spiny neurons from glutamate-induced apoptosis and that Rg5 was protective at micromolar concentration. The other seven ginsenosides tested were not effective or exerted toxic effects in MSN cultures. From further experiments, we suggested that neuroprotective effects of ginsenosides Rb1, Rc, and Rg5 could correlate with their ability to inhibit glutamate-induced Ca(2+) responses in cultured MSN. From these results we concluded that ginsenosides Rb1, Rc, and Rg5 offer a potential therapeutic choice for the treatment of HD and possibly other neurodegenerative disorders. PMID- 19185023 TI - PPTX, a pentraxin domain-containing protein, interacts with the T1 domain of K v 4. AB - Voltage-gated K(+) (K(v)) channels reside as tetramers in the membrane. The events that coordinate folding, trafficking, and tetramerization are mediated by an array of associated proteins and phospholipids whose identification is vital to understanding the dynamic nature of channel expression and activity. An interaction between an A-type K(+) channel, K(v)4.2, and a protein containing a pentraxin domain (PPTX) was demonstrated in the cochlea (Duzhyy et al. [ 2005] J. Biol. Chem. 280:15165-15172). Here, we present results based on fold recognition and homology modeling that revealed the tetramerization (T1) domain of K(v)4.2 as a potential docking site for interacting proteins such as PPTX. By using this model, putative sites were experimentally tested with the yeast two-hybrid system to assay interactions between PPTX and the T1 domain of K(v)4.2 wild type (wt) and mutants (mut). Results showed that amino acid residues 86 and 118 in the T1 domain are essential for interaction, because replacing these negatively charged with neutrally charged amino acids inhibits interactions. Cotransfections of Chinese hamster ovary cells with PPTX and K(v)4.2wt further revealed that PPTX increases K(v)4.2 wt expression in vitro when analyzing total lysates, whereas interactions with K(v)4.2 microt resulted in a decrease. These studies suggest that portions of the T1 domain can act as docking sites for proteins such as PPTX, further underscoring the significance of this domain. PMID- 19185025 TI - Identification of neural cell adhesion molecule L1-derived neuritogenic ligands of the fibroblast growth factor receptor. AB - The neural cell adhesion molecule L1 plays an important role in axon growth, neuronal survival, and synaptic plasticity. We recently demonstrated that the L1 fibronectin type III (FN3) modules interact directly with the fibroblast growth factor (FGF) receptor (FGFR). Sequence alignment of individual L1 FN3 modules with various FGFs suggested that four sequence motifs located in the third and fifth L1 FN3 modules might be involved in interactions with FGFR. The present study found that corresponding synthetic peptides, termed elcamins 1, 2, 3, and 4, bind and activate FGFR in the absence of FGF1. Conversely, in the presence of FGF1, elcamins inhibited receptor phosphorylation, indicating that the peptides are FGFR partial agonists. Elcamins 1, 3, and 4 dose dependently induced neurite outgrowth in cultured primary cerebellar neurons. The neuritogenic effect of elcamins was dependent on FGFR activation, insofar as the effect was abolished by the receptor inhibition. Thus, the identified peptides act as L1 mimetics with regard to activation of FGFR and induction of neurite outgrowth. PMID- 19185024 TI - Spatiotemporal ablation of myelinating glia-specific neurofascin (Nfasc NF155) in mice reveals gradual loss of paranodal axoglial junctions and concomitant disorganization of axonal domains. AB - The evolutionary demand for rapid nerve impulse conduction led to the process of myelination-dependent organization of axons into distinct molecular domains. These domains include the node of Ranvier flanked by highly specialized paranodal domains where myelin loops and axolemma orchestrate the axoglial septate junctions. These junctions are formed by interactions between a glial isoform of neurofascin (Nfasc(NF155)) and axonal Caspr and Cont. Here we report the generation of myelinating glia-specific Nfasc(NF155) null mouse mutants. These mice exhibit severe ataxia, motor paresis, and death before the third postnatal week. In the absence of glial Nfasc(NF155), paranodal axoglial junctions fail to form, axonal domains fail to segregate, and myelinated axons undergo degeneration. Electrophysiological measurements of peripheral nerves from Nfasc(NF155) mutants revealed dramatic reductions in nerve conduction velocities. By using inducible PLP-CreER recombinase to ablate Nfasc(NF155) in adult myelinating glia, we demonstrate that paranodal axoglial junctions disorganize gradually as the levels of Nfasc(NF155) protein at the paranodes begin to drop. This coincides with the loss of the paranodal region and concomitant disorganization of the axonal domains. Our results provide the first direct evidence that the maintenance of axonal domains requires the fence function of the paranodal axoglial junctions. Together, our studies establish a central role for paranodal axoglial junctions in both the organization and the maintenance of axonal domains in myelinated axons. PMID- 19185026 TI - Decreased antioxidant enzyme activity and increased mitochondrial DNA damage in cellular models of Machado-Joseph disease. AB - Machado-Joseph disease (MJD)/spinocerebellar ataxia type 3 (SCA3) is an autosomal dominant neurodegenerative disorder caused by polyglutamine expansion in the ataxin-3 protein that confers a toxic gain of function. Because of the late onset of the disease, we hypothesize that the accumulated oxidative stress or/and defective antioxidant enzyme ability may be contributory factors in the pathogenesis of MJD. In this study, we utilized SK-N-SH and COS7 cells stably transfected with full-length MJD with 78 polyglutamine repeats to examine any alterations in the antioxidant activity. We demonstrated a significant reduction in the ratio of GSH/GSSG and total glutathione content (GSH + 2x GSSG) in mutant MJD cells compared with the wild-type cells under normal or stressful conditions. We also showed that both SK-N-SH-MJD78 and COS7-MJD78-GFP cell lines have lower activities of catalase, glutathione reductase, and superoxide dismutase compared with the wild-type cell lines. In addition, it is known that, when cells are under oxidative stress, the mitochondrial DNA is prone to damage. Our results demonstrated that mitochondrial DNA copy numbers are decreased in mutant cells and SCA3 patients' samples compared with the normal controls. Furthermore, the amount of common mitochondrial DNA 4,977-bp deletion is higher in SCA3 patients compared with that in normal individuals. Overall, mutant ataxin-3 may influence the activity of enzymatic components to remove O(2)(-) and H(2)O(2) efficiently and promote mitochondrial DNA damage or depletion, which leads to dysfunction of mitochondria. Therefore, we suggest that the cell damage caused by greater oxidative stress in SCA3 mutant cells plays an important role, at least in part, in the disease progression. PMID- 19185027 TI - Effects of scoparone on dopamine biosynthesis and L-DOPA-induced cytotoxicity in PC12 cells. AB - The effects of scoparone on dopamine biosynthesis and L-DOPA-induced cytotoxicity in PC12 cells were investigated. PC12 cells treated with scoparone at concentrations of 100-200 microM showed a 128-136% increase in dopamine levels over the course of 24 hr. Scoparone significantly increased the secretion of dopamine into the culture medium. Under the same conditions, the activities of tyrosine hydroxylase (TH) and aromatic L-amino acid decarboxylase (AADC) were enhanced by treatment with 200 microM scoparone for 6-48 hr, but the activity of TH was regulated for a longer period than that of AADC. The intracellular levels of cyclic AMP and Ca(2+) were increased by treatment with 200 microM scoparone. The levels of TH mRNA and the phosphorylation of cyclic AMP-response element binding protein (CREB) were also significantly increased by treatment with 200 microM scoparone. In addition, scoparone at a concentration of 200 microM stimulated the activities of cyclic AMP-dependent protein kinase (PKA), protein kinase C (PKC), and Ca(2+)/calmodulin kinase II (CaMK II). Finally, pretreatment with 200 microM scoparone reduced the cytotoxicity induced by L-DOPA (20-100 microM) at 24 hr. These results suggest that scoparone enhances dopamine biosynthesis by regulating TH activity and TH gene expression, which is mediated by the PKA, CREB, PKC, and CaMK II pathways, and protects cells from L-DOPA induced cytotoxicity by inducing cyclic AMP-PKA systems in PC12 cells. PMID- 19185028 TI - Combined hematopoietic and lentiviral gene-transfer therapies in newborn Twitcher mice reveal contemporaneous neurodegeneration and demyelination in Krabbe disease. AB - This study characterized the therapeutic benefits of combining hematogenous cell replacement with lentiviral-mediated gene transfer of galactosylceramidase (GALC) in Twitcher mice, a bona fide model for Krabbe disease. Bone marrow cells and GALC-lentiviral vectors were administered intravenously without any preconditioning to newborn Twitcher pups before postnatal day 2. Treated Twitchers survived up to 4 months of age. GALC activity remained less than 5% of normal values in the nervous system for the first 2 months after treatment and reached approximately 30% in long-term-surviving mice. Long-term reconstitution of GALC activity in the nervous system was provided primarily by infiltrating macrophages and to a lesser extent by direct lentiviral transduction of neural cells. Treated Twitchers had significant preservation of myelin, with a G-ratio (ratio of the axon diameter to the diameter of the myelinated fiber) in sciatic nerve myelin of 0.75 +/- 0.08 compared with 0.85 +/- 0.10 in untreated mutants. Although treated mutants had improved locomotor activities during their long-term survival, they died with symptoms of progressive neurological degeneration, indistinguishable from those seen in untreated Twitchers. Examination of long lived Twitchers showed that treated mutants were not protected from developing degeneration of axons throughout the neuroaxis. These results suggest that GALC deficiency not only affects myelinating glia but also leads to neuronal dysfunction. The contemporaneous neuropathology might help to explain the limited efficacy of current gene and cell therapies. PMID- 19185030 TI - Hydride-donor abilities of 1,4-dihydropyridines: a comparison with pi nucleophiles and borohydride anions. AB - How are dihydropyridines like indoles? Both groups of compounds have similar nucleophilicity parameters N and are therefore suitable substrates for iminium catalyzed reactions of alpha,beta-unsaturated aldehydes. The N parameters of 1,4 dihydropyridines were derived from the rates of hydride transfer reactions to benzhydrylium ions (see scheme). PMID- 19185031 TI - Differential sensing of serine and tyrosine with aligned CdS nanowire arrays based on pH-dependent photoluminescence behavior. AB - The differential sensing of tyrosine and serine is achieved with well-aligned CdS nanowire arrays by exploring the pH-dependent photoluminescence behavior of the nanowire arrays toward exposure to the two amino acid solutions. The contrasting trend in photoluminescence (PL) intensity with respect to variations in analyte concentration observed at pH 11 served as the check point for the present differential sensing. The application format of the nanowire array is better suited for further sensing device assembly than that of nanocrystal suspensions. PMID- 19185032 TI - Fabrication of monodisperse toroidal particles by polymer solidification in microfluidics. AB - Microdoughnuts: Polymer toroidal particles such as the one shown in the left picture have been prepared by a capillary microfluidic technique. Droplets of polymer solution undergo non-uniform solidification to form the anisotropic polymer particles. By incorporating functional materials inside the polymer network, functional toroidal particles (center and right images) can be tailor made for specific applications such as magnetic actuation. PMID- 19185033 TI - Conflicting observations resolved by a far IR and UV/Vis study of the NO3 radical. AB - By codeposition of NO/Ne and O(2)/Ne mixtures at 6 K, weakly bound complexes between O(2) and NO are formed. They exhibit a strong, structured charge transfer UV band at lambda(max)=275 nm. The UV band disappears during UV irradiation of the neon matrix, while the visible spectrum of the NO(3) radical appears. Simultaneously, the fundamental nu(4) of the NO(3) radical in the X (2)A(2)' ground state is observed in infrared absorption for the first time at 365.6 cm( 1). Its (14/15)N and (16/18)O isotopic shifts reveal strong couplings between the two e'-type modes of NO(3), which are both active in a pseudo-Jahn-Teller interaction with the excited B (2)E' electronic state. The dispute on the vibrational fundamentals of the NO(3) radical is now concluded by the unambiguous assignment of combination bands associated with the fundamental nu(4). Taking into account the observed isotopic shifts and estimated anharmonicities for nu(4) and the most intense IR band of NO(3) at 1492 cm(-1) (nu(3)+nu(4)), the frequency of the so far not observed fundamental nu(3) is estimated to be 1100+/-10 cm(-1). A tentative assignment of the vibronic levels in the IR spectrum in the range from 1000 to 3000 cm(-1) is given. PMID- 19185034 TI - The azulene framework as a novel arene bioisostere: design of potent dopamine D4 receptor ligands inducing penile erection. AB - Blue makes it happen: The non-uniform charge distribution of the blue colored azulene framework is highly suitable for the bioisosteric replacement of bicyclic heteroarene moieties. Showing an analogous binding mode as heterocyclic dopamine D4 receptor-selective lead compounds, the induction of penile erection in rats over a greater range of doses indicates a putative advantage of the rationally developed azulene derivative 2 b over apomorphine. PMID- 19185035 TI - Synthesis and structural characterization of neutral hexacoordinate silicon(IV) complexes with SiO2N4 skeletons. AB - A series of novel neutral hexacoordinate silicon(IV) complexes with SiO(2)N(4) skeletons (compounds 4-8) was synthesized and structurally characterized by single-crystal X-ray diffraction and solid-state NMR spectroscopy ((13)C, (15)N, (29)Si). The silicon(IV) complexes each contain two bidentate monoanionic O,N Schiff base ligands and two cyanato-N or thiocyanato-N ligands. Compounds 4-8 were prepared from Si(NCO)(4) or Si(NCS)(4), whereby the complex formation involved some unexpected chemical transformations of the ligands. PMID- 19185037 TI - Dual amino-functionalised phosphonium ionic liquids for CO2 capture. AB - A series of 20 dual amino-functionalised phosphonium ionic liquids, (3 aminopropyl)tributylphosphonium amino acid salts ([aP(4443)][AA], in which [AA]( ) = [Ala](-), [Arg](-), [Asn](-), [Asp](-), [Cys](-), [Gln](-), [Glu](-), [Gly]( ), [His](-), [Ile](-), [Leu](-), [Lys](-), [Met](-), [Phe](-), [Pro](-), [Ser]( ), [Thr](-), [Trp](-), [Tyr](-) and [Val](-)), has been prepared. Their physicochemical properties, such as density, viscosity, glass transition and thermal decomposition temperatures and conductivity, have been determined. In particular, the [aP(4443)][AA] ionic liquids (ILs) have low glass transition temperatures ranging from -69.7 to -29.6 degrees C and high decomposition temperatures (all above 200 degrees C). The effects of the variation of the structure of [AA](-) on the above physicochemical properties are discussed. Furthermore, the CO(2) absorption of [aP(4443)][Gly], [aP(4443)][Ala], [aP(4443)][Val] and [aP(4443)][Leu], taken as examples, was investigated. It was found that the supported absorption of CO(2) by the [aP(4443)][AA] ILs almost reaches equilibrium within 80 min, the chemical absorption of CO(2) by the [aP(4443)][AA] ILs approaches 1 mol CO(2) per mol ionic liquid (twice that reported before) and the [aP(4443)][AA] ILs can be repeatedly recycled for CO(2) uptake. PMID- 19185036 TI - Spontaneous high-yield production of hydrogen from cellulosic materials and water catalyzed by enzyme cocktails. AB - Cocktail reception: Biohydrogen is produced in high yield from cellulosic materials and water in a one-pot process catalyzed by up to 14 enzymes and one coenzyme. This assembly of enzymes results in non-natural catabolic pathways. These spontaneous reactions are conducted under modest reaction conditions (32 degrees C and atmospheric pressure). PMID- 19185038 TI - Synthesis and conformational analysis of (alpha-D-galactosyl)phenylmethane and alpha-,beta-difluoromethane analogues: interactions with the plant lectin viscumin. AB - (Alpha-D-galactosyl)phenylmethane (1), (alpha- and beta-D galactosyl)(difluoro)phenylmethane (2 and 3) have been prepared and their conformations in solution were described by using a combination of force-field calculations and NMR spectroscopic studies. Galactoside 1 adopts a (4)C(1) chair conformation and an exo anomeric orientation, as is the case for natural alpha galactosides. The X-ray crystal structure of its difluoromethylene derivative 2 similarly shows a (4)C(1) chair conformation. Surprisingly, compound 2 exhibits a different equilibrium between (1)C(4) chair and (1)S(3) skew boat conformations and significant flexibility around the pseudoglycosidic linkage when in solution. The beta-stereoisomer 3 adopts a major (4)C(1) chair conformation. Interestingly, C-galactosides 1, 2, and 3 bind to viscumin (VAA), a galactoside-specific lectin, which is confirmed by NMR experiments and docking calculations. PMID- 19185039 TI - Mechanistic insight into formation of oxo-iron(IV) porphyrin pi-cation radicals from enzyme mimics of cytochrome P450 in organic solvents. AB - Two new models for cytochrome P450 in which the thiolate axial ligand is replaced by a RSO(3)(-) group, form oxo-iron(IV) porphyrin pi-cation radicals as sole oxidation products in "peroxo shunt" reactions independent of the nature of the employed solvent (polar or non-polar) and electronic nature of the porphyrin rings. Although the properties of the solvent and push-pull effects from the porphyrin rings do not affect the mode of the O-O bond cleavage (heterolytic or homolytic) in these models, they strongly affect the rate and mechanism of each reaction step leading to the formation of the high-valent iron intermediates. This article reports the results of mechanistic studies involving the measurements of the rate of oxo-iron(IV) porphyrin pi-cation radical formation from the enzyme mimics of P450 for different oxidant concentration, temperature and pressure in selected organic solvents. Extraction of the appropriate rate constants and activation parameters for the reactions studied enable a detailed discussion of the effects of solvent and electronic nature of the porphyrin rings on the position of the first pre-equilibrium involving formation of the acylperoxo-iron(III) porphyrin intermediate, as well as on the rate of heterolytic O-O bond cleavage leading to the formation of the high-valent iron species. Furthermore, an unusual effect of solvent on the kinetics of oxo iron(IV) porphyrin pi-cation radical formation in methanol is demonstrated and discussed in the present work. PMID- 19185040 TI - Intramolecular axial ligation of zinc porphyrin cores with triazole links within dendrimers. AB - Stable ligation: A series of dendritic porphyrins, such as that depicted in which benzyl ether dendrons were linked to a porphyrin core through 1,2,3-triazole links, was synthesized. Absorption and fluorescence spectra showed a stable axial ligation at the zinc center of the porphyrin core by triazole links in dendritic wedges and indicated that the position of the triazole links strongly affected the stability of the axial ligation within the dendrimer.A series of dendritic porphyrins 7-9 and 12, in which benzyl ether dendrons were linked to a porphyrin core through 1,2,3-triazole links, were synthesized by Cu(I)-catalyzed cycloaddition of azides and alkynes. Absorption and fluorescence spectra showed a stable axial ligation at the zinc center of the porphyrin core by triazole links in dendritic wedges and indicated that the position of the triazole links strongly affected the stability of the axial ligation within the dendrimer. When the porphyrin core was surrounded by aryl ether dendrons having anionic termini and triazole linkers, a significant rate enhancement for photoinduced electron transfer was observed compared with a similar water-soluble dendritic zinc porphyrin lacking triazole linkers. These triazole links constituted a direct pathway within the dendrimer architecture for electron transfer between the zinc porphyrin core and peripheral electron acceptors. PMID- 19185041 TI - Anionic reagents with silicon-containing double bonds. AB - E=Si transfer: Anionic compounds capable of transferring a silicon-containing double bond are reviewed (see figure), particularly reagents with Si=Si moieties (Tip=2,4,6-iPr(3)C(6)H(2), M=Li, Na, K) and their applications towards main-group and transition-metal electrophiles, as well as their reactivity towards organic compounds. A few recently reported derivatives with Si=C (Ad=1-adamantyl) and Si=P moieties are included for completeness.Anionic compounds capable of transferring a silicon double bond are summarized following an introduction to the differences between alkenes and their heavier homologues. The main focus is on reagents with Si=Si moieties and their applications towards main-group and transition-metal electrophiles, as well as their reactivity towards organic compounds, but a few recently reported derivatives with Si=C and Si=P bonds are also included. PMID- 19185042 TI - Enhancing gelation ability of a dendritic gelator through complexation with a polyelectrolyte. AB - A poly(urethane amide) (PUA) dendron with long alkyl chains on its periphery was synthesized and then attached to the backbone of a polyelectrolyte, in which each unit contained a positive charge, by ionizing the carboxyl groups on the apexes of the dendrons to form a dendronized polymer. We found that both the PUA dendron and the dendronized polymer could form organogels in toluene. Interestingly, both the minimum gelation concentration and the gelation time of the dendronized polymer gelator were greatly reduced compared with the dendron alone. Our investigations showed that in the gel phase the intermolecular hydrogen bonding between adjacent dendrons creates similar supramolecular structures in both the dendron and the dendronized polymer gelator, which immobilize solvent molecules by means of interactions between dendrons and solvent molecules. Further studies on the gelation kinetics indicated that the polyelectrolyte backbone plays an important role in prearranging the attached dendritic gelators orderly and quickly into the supramolecular structures through a nucleation-elongation mechanism. Therefore, the gel-forming ability of the dendritic PUA gelator is enhanced by being complexed with the polyelectrolyte. In this work, this positive macromolecular effect is discussed in detail. PMID- 19185043 TI - An extremely simple dibenzopentalene synthesis from 2-bromo-1-ethynylbenzenes using nickel(0) complexes: construction of its derivatives with various functionalities. AB - Nice and easy: A very simple synthesis for dibenzopentalenes, which starts from 1 bromo-2-ethynylbenzenes, has been developed. It uses Ni(0) complexes (see scheme), from which a relatively stable Ni(II) complex as an important intermediate has been isolated. Dibenzopentalenes with various functional groups can be prepared by the procedure, and their electronic properties are consistent with theoretical calculations.An extremely simple dibenzopentalene synthesis from readily available 2-bromo-1-ethynylbenzenes using a nickel(0) complex is described. Although the yields are moderate, the formation of three C-C bonds in a single process and the high availability of the starting materials are important advantages of this reaction. The corresponding aryl-nickel(II) complex as an important intermediate was isolated as relatively stable crystals, and the structure was confirmed by X-ray crystallographic analysis. The high stability of this complex should play a key role in this reaction. The reaction is applicable to the preparation of dibenzopentalenes bearing various functional groups. Their electronic properties are consistent with theoretical calculations. The cyclic voltammograms of these compounds reveal highly amphoteric redox properties. In particular, the electron-donating property of a tetramethoxy derivative is greater than that of oligothiophenes and dibenzodithiophenes and almost comparable to that of pentacene. PMID- 19185044 TI - Synthesis of carbosilane dendrimers containing up to four metal layers. AB - Metallodendrimers are an important class of materials with valuable properties and applications in a large number of areas. Herein, we report a highly efficient route for the synthesis of carbosilane dendrimers containing multimetal layers. The procedure involves the use of heteroditopic and tritopic P,N ligands as connectors of the metal layers. The synthetic strategy is based on the ability of the latter ligands to react selectively with the metal complexes [AuCl(tht)] (tht: tetrahydrothiophene), [{RuCl(2)(p-cymene)}(2)] and [Pd(eta(3)-2 MeC(3)H(4))(cod)](OTf) (cod: 1,5-cyclooctadiene; OTf: triflate). In this way, metallodendrimers containing trimetallic Ru-Au-Pd or tetrametallic Ru-Au-Au'-Pd layers have been formed and characterized. The trimetallic dendrimer 9 can be selectively deconstructed by cleavage of the Ru-N or Au-N bonds by reaction with chloride or iodide salts, respectively. PMID- 19185045 TI - Chemo- and regioselective palladium-catalyzed oxycyclization reactions of allendiols: preparation of five-, six-, and eight-membered cycles. AB - Positive discrimination: Chemo- and regioselective palladium-catalyzed cycloetherification of allendiols, namely beta,gamma- and gamma,delta-allendiols, may occur by judicious choice of palladium-catalyzed conditions owing to their potential ability to discriminate between both nucleophilic sites (see scheme). PMID- 19185046 TI - Asymmetric hydroboration of 1,1-disubstituted alkenes. AB - A breakthrough in the asymmetric hydroboration of notoriously difficult 1,1 disubstituted alkenes using a new family of highly effective hydroboration reagents is described (see scheme). The intermediate boranes can be oxidized to alcohols or used in Suzuki-Miyaura cross-coupling reactions. PMID- 19185047 TI - Radical polymerization of the silene (Me3Si)2Si=CR2 by hydrogen transfer from a trimethylsilyl group. AB - The silene (Me(3)Si)(2)Si=Ad is polymerized to produce a polycarbosilane with an unusual Si-Si-C repeating backbone, rather than the Si-C or Si-Si-C-C units expected for olefinic radical polymerization. The polymer structure and the polymerization mechanism (see scheme) were studied by GPC, EPR, and NMR spectroscopy and by trapping experiments. PMID- 19185048 TI - Aprotinin and the risk of death and renal dysfunction in patients undergoing cardiac surgery: a meta-analysis of epidemiologic studies. AB - PURPOSE: Observational studies have reported conflicting results regarding aprotinin's risk of renal dysfunction and death. A meta-analysis was conducted to summarize results and explain variation of published epidemiologic studies on risks of renal dysfunction and death associated with aprotinin. METHODS: MEDLINE and EMBASE were systematically searched for non-experimental studies that reported risk of renal dysfunction or death with aprotinin use during cardiac surgery in adults. Random-effects meta-analyses were used to pool results across studies for each outcome. Stratified and meta-regression analyses were used to identify sources of heterogeneity. RESULTS: Eleven relevant studies were identified and included in the analysis, including 10 that reported renal dysfunction and seven that reported death. Aprotinin was associated with renal dysfunction (risk ratio (RR), 1.42; 95%CI 1.13-1.79) and long-term mortality (hazard ratio (HR) 1.22; 95%CI 1.08-1.39). Pooled estimates were lower for short term mortality (RR 1.16; 95%CI 0.84-1.58) and renal failure requiring dialysis (RR 1.17; 95%CI 0.99-1.38). Cardiopulmonary bypass (CPB) time, which may be on the causal pathway, was a significant source of heterogeneity, with a 29% increased risk of renal dysfunction for every 10 minute increase in CPB time (p = 0.03). CONCLUSIONS: Despite some studies that reported no association between aprotinin and renal outcomes during cardiac surgery, the totality of epidemiologic evidence indicates an increased risk that cannot be fully explained by need for transfused red blood cells (RBCs). Epidemiologic studies also suggest an increased risk of long-term mortality associated with aprotinin as compared to various comparators used in these studies, although residual confounding cannot be ruled out. PMID- 19185049 TI - De novo T-lymphocyte responses against baculovirus-derived recombinant influenzavirus hemagglutinin generated by a naive umbilical cord blood model of dendritic cell vaccination. AB - Cancer patients and recipients of hematopoietic stem cell transplantation exhibit a negligible response to influenza vaccine. Toward the goal of addressing this issue, we developed an in vitro model of dendritic cell (DC) immunotherapy utilizing DCs generated from naive umbilical cord blood (UCB). UCB DCs were loaded with purified rHA protein and used to stimulate autologous T-lymphocytes. Upon recall with HA-loaded autologous DC, a 4-10-fold increase in the number of IFN-gamma producing T-lymphocytes was observed in comparison to T-cells stimulated with control DCs. Antigen-specific T-cell functionality was determined by (51)Cr lytic assay. Using a peptide library of predicted HA binding epitopes, we mapped an HA-specific, DR15-restricted CD4 T-cell epitope and observed tetramer positive cells. This model demonstrates that HA-specific immune responses might possibly be generated in a de novo fashion and suggests that dendritic cell immunotherapy for the prevention of influenza in populations of immunosuppressed individuals could be feasible. PMID- 19185050 TI - Multi-peptide vaccines vialed as peptide mixtures can be stable reagents for use in peptide-based immune therapies. AB - To date, most peptide-based vaccines evaluated for the treatment of cancer have consisted of one or few peptides. However, as a greater number of peptide antigens become available for use in experimental therapies, it is important to establish the feasibility of combining multi-peptide reagents as individual peptide mixtures. We have found that mixtures of up to 12 peptides can be analyzed accurately for identity, purity, and stability (for at least 5 years) using a combination of high-performance liquid chromatography (HPLC) and mass spectrometry and these complex peptide mixtures have been acceptable for use in human clinical trials. We have also identified some specific concerns for degradation products that should be considered in multi-peptide vaccine preparation and follow-up quality assurance studies. Results from these analyses have implications for changing the way peptide-based vaccines are manufactured and demonstrate that multi-peptide vaccines are reliable reagents for use in peptide-based immune therapies. PMID- 19185051 TI - Hyperbaric oxygen preconditioning reduces ischemia-reperfusion injury by inhibition of apoptosis via mitochondrial pathway in rat brain. AB - This study examined the hypothesis that apoptotic inhibition via mitochondrial pathway was involved in hyperbaric oxygen preconditioning (HBO-PC)-induced neuroprotection on ischemia-reperfusion injury in rat brain. Male Sprague-Dawley rats (250 approximately 280 g, n=144) were divided into control, middle cerebral artery occlusion (MCAO) for 90 min, and HBO-PC plus MCAO groups. HBO-PC was conducted four times by giving 100% oxygen at 2.5 atm absolute (ATA), for 1 h at 12 h intervals for 2 days. At 24 h after the last HBO-PC, MCAO was performed and at 24 h after MCAO, neurological function, brain water content, infarct volume, and cell death were evaluated. Enzymatic activity of capase-3 and -9, and expression of cytochrome c, Bcl-2 and Bax proteins were performed in the samples from hippocampus, ischemic penumbra and core of the brain cortex, respectively. HBO-PC reduced brain edema, decreased infarction volume, and improved neurological recovery. HBO-PC reduced cytoplasm cytochrome c levels, decreased caspase enzyme activity, upregulated the ratio of Bcl-2 and Bax expression, and abated the apoptosis of ischemic tissue. HBO-PC protects brain tissues from ischemia-reperfusion injury by suppressing mitochondrial apoptotic pathways. PMID- 19185052 TI - Transmembrane AMPA receptor regulatory proteins and AMPA receptor function in the cerebellum. AB - Heterogeneity among AMPA receptor (AMPAR) subtypes is thought to be one of the key postsynaptic factors giving rise to diversity in excitatory synaptic signaling in the CNS. Recently, compelling evidence has emerged that ancillary AMPAR subunits-the so-called transmembrane AMPA receptor regulatory proteins (TARPs)-also play a vital role in influencing the variety of postsynaptic signaling. This TARP family of molecules controls both trafficking and functional properties of AMPARs at most, if not all, excitatory central synapses. Furthermore, individual TARPs differ in their effects on the biophysical and pharmacological properties of AMPARs. The critical importance of TARPs in synaptic transmission was first revealed in experiments on cerebellar granule cells from stargazer mice. These lack the prototypic TARP stargazin, present in granule cells from wild-type animals, and consequently lack synaptic transmission at the mossy fibre-to-granule cell synapse. Subsequent work has identified many other members of the stargazin family which act as functional TARPs. It has also provided valuable information about specific TARPs present in many central neurons. Because much of the initial work on TARPs was carried out on stargazer granule cells, the important functional properties of TARPs present throughout the cerebellum have received particular attention. Here we discuss some of these recent findings in relation to the main TARPs and the AMPAR subunits identified in cerebellar neurons and glia. PMID- 19185053 TI - Evolutionary constraints on hub and non-hub proteins in human protein interaction network: insight from protein connectivity and intrinsic disorder. AB - It has been claimed that proteins with more interacting partners (hubs) are structurally more disordered and have a slow evolutionary rate. Here, in this paper we analyzed the evolutionary rate and structural disorderness of human hub and non-hub proteins present/absent in protein complexes. We observed that both non-hub and hub proteins present in protein complexes, are characterized by high structural disorderness. There exists no significant difference in average evolutionary rate of complex-forming hub and non-hub proteins while we have found a significant difference in the average evolutionary rate between hub and non-hub proteins which are not present in protein complexes. We concluded that higher disorderness in complex forming non-hub proteins facilitates higher number of interactions with a large number of protein subunits. High interaction among protein subunits of complex forming non-hub proteins imposes a selective constraint on their evolutionary rate. PMID- 19185054 TI - How scientific is the science in ethnopharmacology? Historical perspectives and epistemological problems. AB - This commentary is based on a general concern regarding the low level of self criticism (-evaluation) in the interpretation of molecular pharmacological data published in ethnopharmacology-related journals. Reports on potentially new lead structures or pharmacological effects of medicinal plant extracts are mushrooming. At the same time, nonsense in bioassays is an increasing phenomenon in herbal medicine research. Only because a dataset is reproducible does not imply that it is meaningful. Currently, there are thousands of claims of pharmacological effects of medicinal plants and natural products. It is argued that claims to knowledge in ethnopharmacology, as in the exact sciences, should be rationally criticized if they have empirical content as it is the case with biochemical and pharmacological analyses. Here the major problem is the misemployment of the concentration-effect paradigm and the overinterpretation of data obtained in vitro. Given the almost exponential increase of scientific papers published it may be the moment to adapt to a falsificationist methodology. PMID- 19185056 TI - Distinct roles for AF-1 and -2 of ER-alpha in regulation of MMP-13 promoter activity. AB - Previous studies have indicated that ER-alpha can influence the activity of the MMP-13 promoter. ER-alpha activity is mediated by two separate transcriptional activation domains (AF-1 and AF-2). The present study focused on analyzing the roles of these domains on the activation of the MMP-13 promoter. Transfection of synoviocytes with an ER-alpha construct lacking the C-terminus AF-2 domain led to significant elevation in MMP-13 promoter activity compared to wild type ER-alpha. Progressive deletions in the N-terminal AF-1 domain led to significant losses in MMP-13 promoter activity. MMP-13 promoter mutagenesis indicated that an AP-1 regulatory site was essential for ER-alpha mutant activity. Thus, both AF-1 and AF-2 domains of ER-alpha are required for regulation of MMP-13 promoter activity. As ER variants and ER related proteins have been implicated in bone and joint disorders, these findings provide understanding of the possible role of ER variants in the development of such conditions. PMID- 19185055 TI - Mechanical ventilation induces diaphragmatic mitochondrial dysfunction and increased oxidant production. AB - Mechanical ventilation (MV) is a life-saving intervention used in patients with acute respiratory failure. Unfortunately, prolonged MV results in diaphragmatic weakness, which is an important contributor to the failure to wean patients from MV. Our laboratory has previously shown that reactive oxygen species (ROS) play a critical role in mediating diaphragmatic weakness after MV. However, the pathways responsible for MV-induced diaphragmatic ROS production remain unknown. These experiments tested the hypothesis that prolonged MV results in an increase in mitochondrial ROS release, mitochondrial oxidative damage, and mitochondrial dysfunction. To test this hypothesis, adult (3-4 months of age) female Sprague Dawley rats were assigned to either a control or a 12-h MV group. After treatment, diaphragms were removed and mitochondria were isolated for subsequent respiratory and biochemical measurements. Compared to control, prolonged MV resulted in a lower respiratory control ratio in diaphragmatic mitochondria. Furthermore, diaphragmatic mitochondria from MV animals released higher rates of ROS in both State 3 and State 4 respiration. Prolonged MV was also associated with diaphragmatic mitochondrial oxidative damage as indicated by increased lipid peroxidation and protein oxidation. Finally, our data also reveal that the activities of the electron transport chain complexes II, III, and IV are depressed in mitochondria isolated from diaphragms of MV animals. In conclusion, these results are consistent with the concept that diaphragmatic inactivity promotes an increase in mitochondrial ROS emission, mitochondrial oxidative damage, and mitochondrial respiratory dysfunction. PMID- 19185057 TI - Free amino acids as indicators of nutritional status of silver bream (Vimba vimba), when using commercial and purified diets. AB - Studies on larval rearing of silver bream (Vimba vimba), a migratory cyprinid fish have addressed on limited scale larval and juvenile rearing using commercial and semipurified diets along with live feeds, such as brine shrimp Artemia nauplii. The objectives of the present study were (1) to determine whether experimental, protein-, peptide-, free amino acid-based diets are adequate for larval silver bream, a stomachless fish, (2) to evaluate whether commercial and purified diets are comparable as the first/exclusive feed for growth and survival of silver bream, and (3) to examine whether free amino acid concentrations in fish body are potential indicators of availability of amino acid sources. We report here the differences in diets acceptance, fish growth and diet utilization in silver bream in comparison to other cyprinid fishes. We specifically address the response in free amino acids in the body to dietary treatments. Experimental diets included: a commercial Aglo Norse feed, casein-gelatin based diet (CG), free amino acid mixture diet (FAA), dipeptide (PP), dipeptide-protein (PP50) based diet, and dipeptide diet without arginine (NoArg). In addition, live Artemia were offered to 3 groups and "fasting" control treatment was included during 3 week long trial. Fish offered Artemia overperformed those offered formulated diets both in terms of mass (80.7+/-26.3 mg) and survival (97.2%). We also indicate that commercial and purified diets are comparable as the first/exclusive feed for growth and survival of silver bream. Our experiment also showed that the whole body free amino acid concentrations of 9 indispensable amino acids (IDAA) out of 10 (His, Thr, Arg, Val, Met, Ile, Leu, Trp, Lys) in the PP50 group was the highest among 7 diet treatments and the totalfree amino acid concentration, total dispensable amino acids (DAA) and total IDAA of the PP50 diet fed fish showed the same trend. This may indicate that diets based on 50% of dipeptides and 50% of protein are adequate for silver bream larvae as they result in similar growth, survival and whole body IDAA concentrations as in fish fed CG (protein-based) diet fed fish. PMID- 19185058 TI - Managing protein flexibility in docking and its applications. AB - Docking, virtual screening and structure-based drug design are routinely used in modern drug discovery programs. Although current docking methods deal with flexible ligands, managing receptor flexibility has proved to be challenging. In this brief review, we present the current state-of-the-art for computationally handling receptor flexibility, including a novel statistical computational approach published recently. We conclude, from a comparison of the different approaches, that a combination of methods is likely to provide the most reliable solution to the problem of finding the right protein conformation for a given ligand. PMID- 19185059 TI - Cellular basis of neurogenesis in the brain of crayfish, Procambarus clarkii: Neurogenic complex in the olfactory midbrain from hatchlings to adults. AB - Neurogenesis in the central olfactory pathway of decapod crustaceans persists throughout life. Here we describe the structural basis of neurogenesis within the olfactory deutocerebrum of the crayfish Procambarus clarkii from hatchlings to adults. Using a proliferation marker and immunostaining, we found that throughout development each hemibrain contains a neurogenic complex consisting of five parts: two proliferation zones, each within the neuronal soma clusters containing local or projection interneurons, a tail of proliferating cells extending from each proliferation zone, and an elongated clump of cells where the two tails meet. The clump of cells comprises two subdivisions joined at a nucleus-free central area. Each subdivision consists of a dense group of clump cells with small, spindle-shaped nuclei and is connected to one of the proliferation zones by a strand of fibrous material encompassing the tail of proliferating cells extending from it. We identify one proliferating cell with a large nucleus in each subdivision as a putative neuroblast. Its daughter cells migrate through the strands to the associated proliferation zones, but in the strand leading to the soma cluster of local interneurons this is masked by local proliferation. We conclude that neurogenesis in the olfactory deutocerebrum of juvenile and adult P. clarkii is based on a few neuroblasts that are associated with unique clumps of cells likely representing stem cell niches. PMID- 19185060 TI - Comprehensive expression analysis of all Wnt genes and their major secreted antagonists during mouse limb development and cartilage differentiation. AB - Wnt signalling plays important roles in patterning and outgrowth of the vertebrate limb. Different mutations in Wnt genes, their antagonists or (co )receptors result in patterning and outgrowth defects as well as chondrocyte and bone phenotypes in mouse and human. Understanding Wnt activity during mouse limb development and chondrogenesis requires a temporal and spatial overview of Wnt signalling key factor expression. Here we present a comparative expression analysis of all 19 Wnt genes and their major secreted antagonists of the Dickkopf (Dkk), Wisp and the secreted frizzled related protein (Sfrp) families during mouse limb development. Our study reveals new domains of expression for Wnt2, Wnt2b, Wnt5b, Wnt6, Wnt7b, Wnt9a, Wnt10a, Wnt10b, Wnt11 and Wnt16, in the limb. We also identified novel expression domains for the Wnt antagonists Sfrp1, Sfrp3, Sfrp5, Wisp1 as well as Dkk2 and Dkk3. We provide a full expression pattern for Wif1 in limb development, for which no limb expression had been documented so far. PMID- 19185061 TI - Anti-inflammatory and anti-asthmatic effects of resveratrol, a polyphenolic stilbene, in a mouse model of allergic asthma. AB - Asthma is an inflammatory disease of the airways, and the current focus in managing asthma is the control of inflammation. Resveratrol (3,4,5 trihydroxystilbene) is a polyphenolic stilbene found in the skins of red fruits, including grapes, that may be responsible for some of the health benefits ascribed to consumption of red wine. We investigated the suppressive effects of resveratrol on asthmatic parameters such as cytokine release, eosinophilia, airway hyperresponsiveness, and mucus hypersecretion, in an OVA-induced allergic mouse model of asthma. Resveratrol significantly inhibited increases in T-helper 2-type cytokines such as IL-4 and IL-5 in plasma and bronchoalveolar lavage fluid (BALF), and also effectively suppressed airway hyperresponsiveness, eosinophilia, and mucus hypersecretion, in the asthmatic mouse model. The efficacy of resveratrol was similar to that of dexamethasone, a glucocorticoid used as a positive control. These results suggest that resveratrol may have applications in the treatment of bronchial asthma. PMID- 19185062 TI - Multiple-modulation effects of Oridonin on the production of proinflammatory cytokines and neurotrophic factors in LPS-activated microglia. AB - Microglial activation has been implicated in many neurological disorders for its inflammatory and/or neurotrophic effects. In this study, we investigated the effects upon activated microglia of Oridonin (Ori), an effective component isolated from Rabdosia rubescens. We pretreated rat primary microglia with or without Ori, then stimulated with lipopolysaccharide (LPS). We found that Ori pretreatment inhibited the release of proinflammatory mediators including nitric oxide (NO), tumor necrosis factor-alpha (TNF-alpha), interleukin-1 beta (IL 1beta) and interleukin-6 (IL-6). Such suppressive effects were accompanied by inhibition of DNA binding activity of the critical transcription factor nuclear factor-kappa B (NF-kappaB). In addition, Ori upregulated the production of the neurotrophic factor, nerve growth factor (NGF). Our findings suggest that Ori may have anti-inflammatory and neuroregulatory effects through modulation of multiple functions of microglia. PMID- 19185063 TI - Imiquimod inhibits the differentiation but enhances the maturation of human monocyte-derived dendritic cells. AB - Imiquimod is a topically used immune response modifier effective in the treatment of genital warts caused by HPV. Its therapeutic effects are believed to be the release of proinflammatory cytokines from the monocyte-macrophage lineage resulting in a cascade of events abating the HPV replication. Dendritic cell maturation and activation have also been found to be induced by imiquimod. We hypothesized that imiquimod may promote the development of DC at all levels of their life cycle. In this study, in vitro cultured monocyte-derived dendritic cells (MoDC) were used to evaluate the effect of imiquimod regarding the modulation of DC differentiation, terminal maturation and their function by phenotypic cell surface molecules expression, cytokine secretion and T cell stimulation in allogeneic system. We demonstrate that imiquimod exerts differential effects on DC biology at different stages of DC development. It inhibits the differentiation of DC, which may indicate a more potent antigen uptake activity. DC maturation is induced by imiquimod with an enhanced antigen presenting activity and IL-12 production. These evidence might be relevant with the clinically proven effectiveness of imiquimod in the treatment of genital warts. PMID- 19185064 TI - The interplay between the geographic distribution of HLA-B27 alleles and their role in infectious and autoimmune diseases: a unifying hypothesis. AB - Due to its strong association with Ankylosing Spondylitis and the other Spondyloarthropathies, the HLA-B27 family of alleles and, in particular, the ancestral HLA-B*2705, has been the object of numerous studies. More recently, some novel interesting features have emerged such as the ability of HLA-B27 to confer resistance to the progression of HIV infection and to promote a spontaneous CD8+ T cell-mediated viral clearance of HCV. The co-occurrence of these protective and pathogenic features suggests a common ground, i.e. to promote a more pronounced immune/inflammatory response leading to an effective clearance of some pathogens on one side and to autoimmunity on the other. This might be due to the antigen presenting properties and/or to the co-inheritance of gene variants that contribute to an altered homeostasis in case of microbial infections or tissue injury. The existence of conserved HLA haplotypes have since long been thought to result from a selective pressure by some pathogens that have edited the immune response genes. The peculiar distribution of the ancestor HLA B*2705 along a latitude-dependent gradient and the opposite distribution of their variants have suggested a correlation with endemic malaria. In this respect, Sardinia, a small Mediterranean island plagued by malaria, represents an interesting laboratory since its population is enriched in conserved HLA haplotypes and several genetic studies have disclosed their correlation with infectious and autoimmune diseases. PMID- 19185065 TI - Identification of regulatory T cells in systemic lupus erythematosus. AB - The concept that regulatory T cells (Treg) play a key role in both development and maintenance of autoimmune response in rheumatic diseases is well accepted. In recent years, several studies analyzed Treg cell phenotype and function in systemic lupus erythematosus (SLE), the prototypical systemic autoimmune disorder in humans. Although qualitative and/or quantitative abnormalities of Treg cells have been shown, data are often conflicting. This may depend on the selection of patients with different degrees of disease activity or on immunosuppressive treatments that can alter Treg cell findings. Among several proposed surface or intracellular Treg cell markers, CD25 at high level of expression and the transcription factor Foxp3 are the two most investigated in SLE. Despite the glucocorticoid-induced TNF receptor-related protein (GITR) represents a reliable phenotypic marker of murine Treg cells, little is known about its role in humans, in particular in the course of systemic autoimmune disorders. Preliminary data seems to suggest that this marker may represent a good tool to identify cell populations included within Treg cell subsets. PMID- 19185066 TI - Membrane-bound proteinase 3 and its receptors: relevance for the pathogenesis of Wegener's Granulomatosis. AB - Wegener's Granulomatosis (WG) is a life-threatening autoimmune disease. A pathogenic role for anti-neutrophil cytoplasmic autoantibodies (ANCAs) by inducing necrotizing damage to the vessel wall has been strongly suggested by in vitro and in vivo experimental data. Proteinase 3 (PR3), a serine protease mainly stored in the azurophilic granules of neutrophils, has been identified as a major ANCA-antigen in WG. Elevated expression levels of membrane-bound PR3 (mPR3) has been observed in WG and some other chronic inflammatory diseases, suggesting a pathogenic role of mPR3 by allowing interaction with PR3-ANCA. Recent studies revealed CD177 as a receptor for mPR3 on the neutrophil membrane. However, we recently showed that CD177 negative neutrophils also express mPR3 and are susceptible to PR3-ANCA induced neutrophil activation. Therefore, it is of interest to further investigate the functional consequences of binding of mPR3 to CD177, to explore other binding partners for mPR3 on the neutrophil membrane, and to study the relevance of colocalization of these molecules for disease pathogenesis. This review gives updated information on the mechanism of mPR3 expression and the relevance of colocalization of mPR3 with other molecules on the neutrophil membrane for the pathophysiological events occurring in WG. PMID- 19185067 TI - Minimum information requirements: neither bandits in the Attic nor bats in the belfry. AB - In respose to de Marco and colleagues 'Minimum Information for Protein Functional Evaluation,' I observe that they run the risk of imposing a Procrustean solution to the problem of rigorously reporting experimental results involving recombinant proteins. While they rightly identify a need for experimental reporting standards defined by their scientific community, they wrongly assign this role to 'Minimum information' guidelines. In my response I clarify the role and development processes of minimum information standards, and relate a similar experience that we had in the ProteomeBinders Consortium, that led to a distinction between minimum information guidelines for describing a binder and its efficiency as a molecular tool, and the need to define community standards for experimental binder assessment. PMID- 19185068 TI - Disulphide proteomes and interactions with thioredoxin on the track towards understanding redox regulation in chloroplasts and cyanobacteria. AB - Light-dependent disulphide/dithiol exchange catalysed by thioredoxin is a classical example of redox regulation of chloroplast enzymes. Recent proteome studies have mapped thioredoxin target proteins in all chloroplast compartments ranging from the envelope to the thylakoid lumen. Progress in the methodologies has made it possible to identify which cysteine residues interact with thioredoxin and to tackle membrane-bound thioredoxin targets. To date, more than hundred targets of thioredoxin and glutaredoxin have been found in plastids from Arabidopsis, spinach, poplar and Chlamydomonas reinhardtii. Thioredoxin-mediated redox control appears to be a feature of the central pathways for assimilation and storage of carbon, sulphur and nitrogen, as well as for translation and protein folding. Cyanobacteria are oxygenic photosynthetic prokaryotes, which presumably share a common ancestor with higher plant plastids. As in chloroplasts, cyanobacterial thioredoxins receive electrons from the photosynthetic electron transport, and thioredoxin-targeted proteins are therefore highly interesting in the context of acclimation of these organisms to their environment. Studies of the unicellular model cyanobacterium Synechocystis sp. PCC 6803 revealed 77 thioredoxin target proteins. Notably, the functions of all these thioredoxin targets highlight essentially the same processes as those described in chloroplasts suggesting that thioredoxin-mediated redox signalling is equally significant in oxygenic photosynthetic prokaryotes and eukaryotes. PMID- 19185069 TI - Optimizing management strategies in patients with chronic hepatitis B. Introduction. PMID- 19185070 TI - Diagnosis of chronic hepatitis B and the implications of viral variants and mutations. AB - Chronic infection with the hepatitis B virus (HBV) is a major cause of morbidity and mortality worldwide, necessitating accurate and timely diagnosis of infected patients coupled with optimal treatment strategies. Although the prevalence of HBV infection in the United States is low owing to the implementation of universal vaccination, growing immigration from areas where infection is more endemic means that prevalence is forecast to increase. Healthcare providers must be active in providing low-cost screening (hepatitis B surface antigen identification) and vaccination programs for high-risk communities, such as Asian Americans, with linked specialist referral schemes for patients found to carry the virus. Serologic technologies and improved nucleic acid testing techniques generate important information about the stage of disease, viral load, and disease subtype, including the presence of precore and core promoter variants that provide prognostic indicators and can guide patient management. Serial DNA monitoring is playing the major role in the assessment of therapeutic response and steering treatment approaches. More research is needed to further clarify the significance of HBV variants and their relation to therapeutic agents and strategies. PMID- 19185071 TI - Assessment of the newly diagnosed patient with chronic hepatitis B virus infection. AB - An expanding population of patients with chronic hepatitis B (CHB) infection, including immigrants from endemic countries, are being identified in the United States. All newly diagnosed patients with CHB should be evaluated for initial disease severity and potential antiviral treatment options. Following a history and physical examination, routine laboratory measurements along with serum markers of hepatitis B virus (HBV) replication should be obtained. A liver biopsy provides important information regarding disease activity and stage and can help identify suitable candidates for antiviral therapy. Untreated patients with CHB should undergo periodic monitoring of serum aminotransferase and HBV DNA levels, as well as surveillance for hepatocellular carcinoma. Avoidance of hepatotoxic medications and immunosuppressants, as well as oral antiviral prophylaxis in selected clinical circumstances, is recommended for all patients with CHB. Behavioral modifications and education can help minimize the risk of inadvertent HBV transmission to close contacts. Although HBV is an entirely preventable disease via the use of a safe and highly effective vaccine, continued morbidity and mortality will likely be encountered due to the delay in diagnosis and treatment of the large number of patients with chronic HBV in the United States and worldwide. PMID- 19185072 TI - Demography and presentation of chronic hepatitis B virus infection. AB - Currently, >350 million people worldwide are affected by chronic infection with the hepatitis B virus (HBV). Chronic infection may cause cirrhosis and hepatocellular carcinoma; HBV infection is responsible for 328,000 cancer deaths per year. In areas of high HBV endemicity, most infections occur early in life; infected children do not mount an effective immune response and exhibit immune tolerance, so that the risk of chronic infection is high. In areas of low endemicity, infections tend to be in adults within defined risk groups, and the risk of chronicity is much lower. Population migration from areas of high endemicity to areas of low endemicity is creating pockets of HBV infection in areas of low general prevalence, necessitating improved efforts to screen, vaccinate, and treat. Chronic HBV infection is a complicated, nonlinear disease with a variable course of progression; predictors of progression include the duration of time in the immunoactive phase of disease that follows the immune tolerant phase when hepatocytes are attacked. Additionally, the duration of a high viremic state, with ongoing clinical hepatitis and possibly concurrent infections (e.g., hepatitis C, human immunodeficiency virus), influence outcome. Targeted vaccination of high-risk groups has many limitations. Universal childhood vaccination to prevent chronic infection and its sequelae is the only approach that will lead to the global elimination of chronic HBV infection. PMID- 19185073 TI - Therapeutic strategies for chronic hepatitis B virus infection in 2008. AB - Reducing progression of disease and preventing development of hepatocellular carcinoma in patients with chronic hepatitis B (CHB) is best achieved through durable suppression of viral replication without the development of resistance. Suitability of a patient for therapy depends on several factors, notably hepatitis B e antigen (HBeAg) status, serum alanine aminotransferase (ALT) level, and serum HBV DNA level. Interferon therapy can produce durable responses (HBeAg clearance) in approximately one third of patients with HBeAg-positive disease, but has limited use in HBeAg-negative patients owing to relapse after discontinuation of therapy. Oral antiviral agents have superior tolerability and ease of administration, but the long-term treatment that may be necessary can be associated with the development of resistance. The high propensity for development of resistance to lamivudine and telbivudine means that adefovir and entecavir, and where available tenofovir, are now considered the preferred oral agents. Monitoring patients during therapy enables assessment of response and adherence, such that treatment can be modified accordingly; genotypic testing in cases of virologic breakthrough is important to identify presence of resistant strains, which may be found months or years before elevation of serum ALT. In cases of confirmed antiviral resistance, treatment should be promptly adjusted either by switching to another agent, or more commonly, adding a second agent, depending on the circumstances. Long-term studies indicate that effective suppression of viral replication improves clinical outcome, with improvement of liver histology and, it is assumed, a decreased risk for development of hepatocellular carcinoma. PMID- 19185074 TI - Implementing evidenced-based practice guidelines for the management of chronic hepatitis B virus infection. AB - In the past 2 years, evidenced-based guidelines and statements on screening, diagnosis, and management of hepatitis B virus (HBV) from several organizations and experts have been published. The purpose of this article is to take the recommendations from these documents and help guide clinicians--whether they are primary care providers, hepatologists, gastroenterologists, or public health workers--about how to incorporate these guidelines into practice. The first task is for all providers to be involved in identifying persons with chronic HBV infection (CHB). New recommendations from the Centers for Disease Control and Prevention (CDC) advocate screening for the HBV in those at highest risk, especially persons from countries where HBV is endemic. Using information from the clinical and laboratory evaluation of the patient infected with HBV, especially the hepatitis B e antigen/antibody status, clinicians can classify the patient into 1 of the 4 phases of HBV infection. Because CHB is a dynamic process and patients can move from inactive to active infection status, and vice versa, all patients must be followed with alanine aminotransferase (ALT) and aspartate aminotransferase monitoring every 3 to 12 months for life. Those with elevated ALT and HBV DNA levels (>2,000 IU/mL) should be referred to a specialist for evaluation for possible treatment. Patients selected for antiviral therapy with nucleoside analogues should be followed every 3 to 6 months to detect emergence of antiviral resistance to the agent chosen. In addition, alpha-fetoprotein should be tested and ultrasound performed on all men aged >40 years and women >50 years of age to detect any hepatocellular carcinoma (HCC) in an early stage. Algorithms are included for primary care providers providing information on initial evaluation and management and referral of persons with CHB, and for specialists evaluating and treating HBV. Implementing steps to identify, follow, refer, and treat appropriately persons with CHB infection by all primary care and specialist healthcare providers can have a major impact on reducing the occurrence of HCC and cirrhosis in infected persons. PMID- 19185075 TI - Reduced efficacy of rosuvastatin by St. John's Wort. PMID- 19185076 TI - Unusual case of hookworm presenting as acute surgical abdomen. PMID- 19185077 TI - More study needed on morning erections and erectile dysfunction. PMID- 19185078 TI - Can the gender imbalance in mitral stenosis prevalence be mitigated? PMID- 19185079 TI - Hypertension and women: a worldwide project. PMID- 19185080 TI - The American Journal of Medicine blog: an invitation to participate. PMID- 19185081 TI - Premed requirements: the time for change is long overdue! PMID- 19185082 TI - Risk stratification in non-ST-segment elevation acute coronary syndromes: troponin alone is not enough. PMID- 19185083 TI - From randomized controlled trials to observational studies. AB - Randomized controlled trials are considered the gold standard in the hierarchy of research designs for evaluating the efficacy and safety of a treatment intervention. However, their results can have limited applicability to patients in clinical settings. Observational studies using large health care databases can complement findings from randomized controlled trials by assessing treatment effectiveness in patients encountered in day-to-day clinical practice. Results from these designs can expand upon outcomes of randomized controlled trials because of the use of larger and more diverse patient populations with common comorbidities and longer follow-up periods. Furthermore, well-designed observational studies can identify clinically important differences among therapeutic options and provide data on long-term drug effectiveness and safety. PMID- 19185084 TI - Atopic dermatitis. PMID- 19185085 TI - Anticoagulation: a pathway to clinical effectiveness. PMID- 19185086 TI - Feverish, jaundiced. PMID- 19185087 TI - Beneath the surface. PMID- 19185088 TI - Pebbly skin. PMID- 19185089 TI - A dangerous secret. PMID- 19185090 TI - A 40-day siege. PMID- 19185091 TI - Predictors and implications of Q-waves in ST-elevation acute coronary syndromes. AB - BACKGROUND: Q-waves in ST-elevation acute coronary syndromes carry adverse implications. We sought to determine the frequency, predictors, and implications of Q-waves in the current era that includes primary percutaneous coronary interventions. METHODS: There were 14,916 patients evaluated in a multicenter observational study. They presented with ST-elevation acute coronary syndromes between 1999 and 2006. Clinical variables were compared between patients with versus without presenting Q-waves, with an additional comparison in the latter group between those with versus without subsequent development of Q-waves. RESULTS: ST-elevation myocardial infarction occurred in 88.6% of patients. Q waves were present on the initial electrocardiogram in 3929 patients and developed later in an additional 3085 patients. The incidence of Q-waves at presentation or during hospitalization decreased from 61% to 39% between 1999 and 2006 (linear trend P<.001). Both presenting and subsequent Q-waves were associated with greater likelihood of coronary occlusions and higher cardiac marker elevations (P <.001). Multivariate analysis showed that presenting Q-waves were associated with male sex (odds ratio [OR] 1.28), increased age (OR 1.06 per 5 years), diabetes (OR 1.26), smoking (OR 1.11), chronic aspirin (OR 0.79), acute aspirin (OR 0.87), other chronic cardiac medications (OR 0.80), prior heart failure (OR 0.67), and prior coronary artery disease (OR 0.61). Presenting Q waves were independently associated with increased in-hospital mortality (OR 1.46), but Q-waves at presentation or during hospitalization did not impact 6 month mortality. CONCLUSIONS: Q-waves in ST-elevation acute coronary syndromes are decreasing in incidence. Q-waves are a major determinant of in-hospital mortality, and targeted interventions should be directed to these high-risk patients. PMID- 19185092 TI - The impact of revascularization on mortality in patients with nonacute coronary artery disease. AB - BACKGROUND: Although early revascularization improves outcomes for patients with acute coronary syndromes, the role of revascularization for patients with nonacute coronary artery disease is controversial. The objective of this meta analysis was to compare surgical or percutaneous revascularization with medical therapy alone to determine the impact of revascularization on death and nonfatal myocardial infarction in patients with coronary artery disease. METHODS: The Medline and Cochrane Central Register of Controlled Trials databases were searched to identify randomized trials of coronary revascularization (either surgical or percutaneous) versus medical therapy alone in patients with nonacute coronary disease reporting the individual outcomes of death or nonfatal myocardial infarction reported at a minimum follow-up of 1 year. A random effects model was used to calculate odds ratios (OR) for the 2 prespecified outcomes. RESULTS: Twenty-eight studies published from 1977 to 2007 were identified for inclusion in the analysis; the revascularization modality was percutaneous coronary intervention in 17 studies, coronary bypass grafting in 6 studies, and either strategy in 5 studies. Follow-up ranged from 1 to 10 years with a median of 3 years. The 28 trials enrolled 13,121 patients, of whom 6476 were randomized to revascularization and 6645 were randomized to medical therapy alone. The OR for revascularization versus medical therapy for mortality was 0.74 (95% confidence interval [CI], 0.63-0.88). A stratified analysis according to revascularization mode revealed both bypass grafting (OR 0.62; 95% CI, 0.50-0.77) and percutaneous intervention (OR 0.82; 95% CI, 0.68-0.99) to be superior to medical therapy with respect to mortality. Revascularization was not associated with a significant reduction in nonfatal myocardial infarction compared with medical therapy (OR 0.91; 95% CI, 0.72-1.15). CONCLUSION: Revascularization by coronary bypass surgery or percutaneous intervention in conjunction with medical therapy in patients with nonacute coronary artery disease is associated with significantly improved survival compared with medical therapy alone. PMID- 19185093 TI - Do income level and race influence survival in patients receiving hemodialysis? AB - BACKGROUND: Residence in a lower-income area has been associated with higher mortality among patients receiving dialysis. We sought to determine whether these differences persist and whether the effect of income-area on mortality is different for African Americans versus patients of other races. METHODS: We evaluated relationships between lower- and higher-income versus middle-income area residence and mortality to 5 years after adjusting for differences in baseline clinical, dialysis facility, and socioeconomic characteristics in 186,424 adult patients with end-stage renal disease initiating hemodialysis at stand-alone facilities between 1996 and 1999. We also compared mortality differences between race and income level groups using non-African Americans residing in middle-income areas as the reference group. RESULTS: Patients with end-stage renal disease who reside in lower-income areas were younger and more frequently African American. After adjustment, there were no mortality differences among income level groups. However, African Americans in all income level groups had lower adjusted mortality compared with the reference group (lower-income hazard ratio [HR]=0.771, 95% confidence interval [CI], 0.736-0.808; middle-income HR=0.755, 95% CI, 0.730-0.781; higher-income HR=0.809, 95% CI, 0.764-0.857), whereas adjusted mortality was similar among non-African-American income level groups (lower-income HR=1.019, 95% CI, 0.976-1.064; higher-income HR=1.003, 95% CI, 0.968-1.039). CONCLUSION: Adjusted survival for patients receiving hemodialysis in all income areas was similar. However, this result masks the paradoxically higher survival for African American versus patients of other race and demonstrates the need to adjust for differences in demographic, clinical, provider, and socioeconomic status characteristics. PMID- 19185094 TI - Diagnostic usefulness of a T-cell-based assay for extrapulmonary tuberculosis in immunocompromised patients. AB - BACKGROUND: The low reactivity of the tuberculin skin test limits its clinical use in immunocompromised patients with extrapulmonary tuberculosis. A recently developed T-cell-based assay for diagnosing tuberculosis infection gave promising results. However, there were few data on the usefulness of this assay for diagnosing extrapulmonary tuberculosis in immunocompromised patients. METHODS: All adult patients with suspected extrapulmonary tuberculosis were prospectively enrolled at 2 university-affiliated hospitals over an 18-month period. In addition to the conventional tests for diagnosing extrapulmonary tuberculosis, enzyme-linked immunospot (ELISPOT) assay for the interferon-gamma-producing T cell response to early secretory antigenic target-6 and culture filtrate protein 10 was performed. The final diagnoses in patients with suspected extrapulmonary tuberculosis were classified by clinical category. RESULTS: There were 179 patients with suspected extrapulmonary tuberculosis enrolled: 59 (33%) were classified as immunocompromised. Of the 179 patients, 75 (42%) were classified as extrapulmonary tuberculosis, including 56 confirmed tuberculosis plus 19 probable tuberculosis, and 97 (54%) were classified as not tuberculosis. The remaining 7 (4%) had possible tuberculosis and were excluded from the final analysis. The tuberculin skin test (induration size >or=10 mm) was less sensitive in immunocompromised patients (38%; 95% confidence interval [CI], 19%-59%) than in immunocompetent patients (69%; 95% CI, 54%-81%, P=.01). In contrast, the ELISPOT assay retained a high sensitivity: (88%; 95% CI, 68%-97%) in immunocompromised patients compared with 96% (95% CI, 87%-100%) in immunocompetent patients (P=.32). CONCLUSION: The immunosuppressive condition does not affect the diagnostic sensitivity of the ELISPOT assay for extrapulmonary tuberculosis. PMID- 19185095 TI - Sources of satisfaction: a second administration of the program director satisfaction survey. PMID- 19185098 TI - Inherited thrombophilia and preeclampsia: is the evidence beginning to congeal? PMID- 19185097 TI - Prospective evidence that lactation protects against cardiovascular disease in women. PMID- 19185099 TI - Umbilical cord occlusion for selective feticide in complicated monochorionic twins: a systematic review of literature. AB - The aim of this study was to review literature concerning selective feticide (SF) in monochorionic pregnancies complicated with twin-twin transfusion syndrome, twin reversed arterial perfusion, severe malformation, and discordant growth. The remaining twins' outcomes were stratified for indication and surgical technique. Three hundred forty-five cases of SF were reviewed. Premature rupture of membranes was described in all the procedures and complicated 76/345 (22%) pregnancies within 4 postoperative weeks (59%) or later (41%; P = .52). Fetal demise accounted for 51/345 (15%), more frequently within 2 postoperative weeks (79%) than thereafter (21%; P = .004; odds ratio [OR], 6.12; 95% confidence interval [CI], 1.81-20.70). Improved survival rate was achieved in surgeries after 18 weeks (89%) than earlier (69%; P = .02; OR, 0.28; 95% CI, 0.10-0.80) without differences for indication. Survivors were 86% after radiofrequency ablation, 82% after bipolar cord coagulation, 72% after laser cord coagulation, 70% after cord ligation. In spite of favorable outcomes, the optimal surgical approach remains undetermined. PMID- 19185100 TI - Enhanced estrogen-induced proliferation in obese rat endometrium. AB - OBJECTIVE: We tested the hypothesis that the proliferative estrogen effect on the endometrium is enhanced in obese vs lean animals. STUDY DESIGN: Using Zucker fa/fa obese rats and lean control, we examined endometrial cell proliferation and the expression patterns of certain estrogen-regulated proproliferative and antiproliferative genes after short-term treatment with estradiol. RESULTS: No significant morphologic/histologic difference was seen between the obese rats and the lean rats. Estrogen-induced proproliferative genes cyclin A and c-Myc messenger RNA expression were significantly higher in the endometrium of obese rats compared with those of the lean control. Expression of the antiproliferative gene p27Kip1 was suppressed by estrogen treatment in both obese and lean rats; however, the decrease was more pronounced in obese rats. Estrogen more strongly induced the antiproliferative genes retinaldehyde dehydrogenases 2 and secreted frizzled-related protein 4 in lean rats but had little or no effect in obese rats. CONCLUSION: Enhancement of estrogen-induced endometrial proproliferative gene expression and suppression of antiproliferative gene expression was seen in the endometrium of obese vs lean animals. PMID- 19185101 TI - Comparison of APTIMA Trichomonas vaginalis transcription-mediated amplification to wet mount microscopy, culture, and polymerase chain reaction for diagnosis of trichomoniasis in men and women. AB - OBJECTIVE: We evaluated the performance characteristics of APTIMA Trichomonas vaginalis (ATV) transcription-mediated amplification (TMA) for diagnosis of T vaginalis (TV) infection from female vaginal swab, endocervical swab, and urine specimens and from male urethral swab and urine specimens. Performance of ATV TMA was compared with wet mount microscopy, culture, and polymerase chain reaction (PCR). STUDY DESIGN: In all, 296 female and 298 male subjects who attended the Jefferson County Health Department sexually transmitted diseases clinic were enrolled in the study and provided specimens for each test. Results were analyzed using 3 interpretative algorithms. RESULTS: For women, vaginal swab ATV TMA was significantly more sensitive than wet mount or culture. In male subjects, urethral swab ATV TMA was significantly more sensitive than culture or PCR. CONCLUSION: ATV TMA provides a sensitive, commercially available nucleic acid amplification test for improved diagnosis of TV in male and female patients. PMID- 19185104 TI - Surgery is different: a response to the IOM report. PMID- 19185102 TI - Fetal renal artery impedance as assessed by Doppler ultrasound in pregnancies complicated by intraamniotic inflammation and preterm birth. AB - OBJECTIVE: The objective of the study was to evaluate the fetal renal artery impedance in the context of inflammation-associated preterm birth. STUDY DESIGN: We conducted a prospective Doppler assessment of the fetal renal artery impedance in 70 singleton fetuses. The study group consisted of 56 premature fetuses (median, 28.1 [interquartile range, 25.3-30.6] weeks at enrollment). Gestational age (GA) reference ranges were generated based on fetuses with uncomplicated pregnancies (n = 14). Doppler studies included renal artery pulsatility index (PI), resistance index (RI), systolic/diastolic (S/D) ratio, and presence or absence of end-diastolic blood flow. Proteomic profiling (surface-enhanced laser desorption ionization time-of-flight) was used for assessment of intraamniotic inflammation and biomarker peak corresponding to beta2-microglubin. Data were interpreted in relationship to amniotic fluid index (AFI), cord blood interleukin (IL)-6 and erythropoietin (EPO) levels. The cardiovascular and metabolic profiles of the neonates were investigated in the first 24 hours of life. RESULTS: Fetuses delivered by mothers with intraamniotic inflammation had higher cord blood IL-6 but not EPO levels. Fetal inflammation did not affect either renal artery PI, RI, S/D ratio, or end-diastolic blood flow. Neonates delivered in the context of intraamniotic inflammation had higher serum blood urea nitrogen levels, which correlated significantly with AF IL-6 levels. The renal artery RI and SD ratio were inversely correlated with the AFI independent of GA, cord blood IL-6, and status of the membranes. CONCLUSION: The fetus is capable of sustaining normal renal artery impedance despite inflammation. Resistance in the renal vascular bed affects urine output independent of inflammation. PMID- 19185105 TI - Presidential Address: Dr. Organ, how are we doing? PMID- 19185106 TI - The Asa Yancey Lecture: swimming against the current: building strength through adversity. PMID- 19185107 TI - 17beta-estradiol mediates protection against microvascular endothelial cell hyperpermeability. AB - BACKGROUND: Previous work from our laboratory demonstrated the involvement of "intrinsic" mitochondrial apoptotic signaling in vascular hyperpermeability. The objective of this study was to determine if 17beta-estradiol, a known inhibitor of apoptosis, would attenuate microvascular endothelial cell hyperpermeability. METHODS: Rat lung microvascular endothelial cell monolayers were treated with 17beta-estradiol or estrogen-receptor antagonist ICI 182780 after transfection with BAK peptide (5 microg/mL). Fluorescein isothiocyanate (FITC)-albumin was used to determine the change in permeability. Mitochondrial reactive oxygen species (ROS) formation and transmembrane potential were determined using 123 dihydrorhodamine and JC-1, respectively. Cytosolic cytochrome c levels and caspase-3 activity were determined using enzyme-linked immunosorbent assay and fluorometric assay respectively. RESULTS: 17beta-estradiol (10 nm) attenuated BAK induced hyperpermeability (P < .05), ROS formation, cytochrome c release, and caspase-3 activation. The estrogen receptor antagonist ICI 182780 blocked the protective effect of 17beta-estradiol on hyperpermeability (P < .05). CONCLUSIONS: 17beta-estradiol attenuates BAK-induced hyperpermeability in rat lung microvascular endothelial cells by way of an estrogen-receptor mediated pathway. PMID- 19185108 TI - Severe traumatic brain injury: is there a gender difference in mortality? AB - BACKGROUND: Emerging evidence suggests that male and female nervous systems respond differently to traumatic brain injury (TBI). The objective of this study was to examine outcomes between the sexes after TBI. PATIENTS AND METHODS: A retrospective review of all severe TBI patients admitted between January and December 2005 was performed. Isolated severe TBI was defined as a head abbreviated injury score greater than 3 with an abbreviated injury score of 3 or less for other anatomic regions. The population was stratified into age subgroups (<14 y, 14-44 y, 45-54 y, and > or =55 y). Logistic regression was used to identify independent predictors of mortality. RESULTS: A total of 1,807 TBI patients were admitted. The mortality was significantly higher for women (43.2% vs 36.2%, P < .01) with an adjusted odds ratio of 1.4 (95% confidence interval, 1.1-1.9, P < .05). After stratification, only women age 55 and older had a significant difference in mortality (odds ratio, 1.71; 95% confidence interval, 1.11-2.62, P = .02). CONCLUSIONS: Female sex (particularly those age > or =55 y) is associated independently with higher mortality in isolated severe TBI. This increased mortality of postmenopausal women after isolated TBI may suggest a hormonal influence and warrants further investigation. PMID- 19185109 TI - Clinical utility of breast-specific gamma imaging for evaluating disease extent in the newly diagnosed breast cancer patient. AB - BACKGROUND: Breast-specific gamma imaging (BSGI) is a functional imaging modality that has comparable sensitivity but superior specificity compared with magnetic resonance imaging, yielding fewer false-positive results and thereby improving clinical management of the newly diagnosed breast cancer patient. METHODS: A retrospective review was performed from 2 community-based breast imaging centers of newly diagnosed breast cancer patients in whom BSGI was performed as part of the imaging work-up. RESULTS: A total of 138 patients (69 invasive ductal carcinoma, 20 invasive lobular carcinoma, 32 ductal carcinoma in situ, and 17 mixtures of invasive ductal carcinoma, invasive lobular carcinoma, or ductal carcinoma in situ and other) were reviewed. Twenty-five patients (18.1%) had a positive BSGI study at a site remote from their known cancer or more extensive disease than detected from previous imaging. Fifteen patients (10.9%) were positive for a synchronous or more extensive malignancy in the same or contralateral breast. Five patients had benign findings on pathology, 5 benign on ultrasound follow-up (false-positive rate, 7.2%). Findings converted 7 patients to mastectomy, 1 patient to neoadjuvant chemotherapy, and 7 patients were found to have previously undetected contralateral cancer. The positive predictive value for BSGI was 92.9%. CONCLUSIONS: BSGI detected additional or more extensive malignancy in the same or contralateral breast in 10.9% of newly diagnosed breast cancer patients. Only 7.2% incurred an additional work-up. BSGI provides accurate evaluation of remaining breast tissue in newly diagnosed breast cancer patients with few false-positive readings. PMID- 19185110 TI - Fournier's gangrene: vacuum-assisted closure versus conventional dressings. AB - BACKGROUND: Fournier's gangrene is a fulminant and destructive inflammation of the scrotum, penis, and perineum. The objective of this study was to compare 2 different approaches to wound management after aggressive surgical debridement. METHODS: Data from 35 patients with Fournier's gangrene were prospectively collected (1996-2007). Once the patients were stabilized following surgery, they were treated with either daily antiseptic (polyhexanide) dressings (group I, n = 16) or vacuum-assisted closure (VAC) therapy (group II, n = 19). RESULTS: The mean age of the patients was 58.2 years in group I and 57.2 years in group II. In both groups, the most common predisposing conditions were diabetes mellitus, chronic alcoholism, and obesity. Escherichia coli, streptococcal species, Pseudomonas aeruginosa, and Staphylococcus aureus were the most frequently isolated organisms. Length of hospital stay was 27.8 days +/- 27.6 days (mortality: 37.5%) in group I and 96.8 days +/- 77.2 days (mortality: 5.3%) in group II. Enterostomies were performed in 43.8% of group I patients and in 89.5% of group II patients. CONCLUSIONS: VAC was associated with significantly longer hospitalization and lower mortality. A partial explanation is that some patients with severe sepsis died within the first 3 days after admission and thus could not undergo vacuum therapy. Since our clinical experience has shown that vacuum dressings are particularly effective in the management of large wounds, we use VAC primarily for this indication despite the considerable material requirements involved. PMID- 19185111 TI - The availability of circulating blood volume values alters fluid management in critically ill surgical patients. AB - BACKGROUND: This study evaluated whether commercially available blood volume measurements in critically ill surgical patients altered fluid management. METHODS: Patients admitted to the surgical intensive care unit of a tertiary care teaching hospital were prospectively evaluated. The frequency of changes in fluid management when results of blood volume measurements were available was determined. RESULTS: In a pilot study, the frequency of instances when measurement of blood volume would have altered fluid management was statistically significant (P = .0003). In 40 subsequent patients, treatment change occurred in 36% of instances when blood volume results were obtained (P < .001). In the majority, no immediate qualitative change in clinical status occurred, with a desirable clinical response in 39% and no negative treatment responses (P < .001). CONCLUSIONS: Blood volume measurements may assist in the management of critically ill surgical patients by providing a direct measure of intravascular volume. Further studies are warranted to determine its effect on outcome. PMID- 19185112 TI - Novel approaches for selenium speciation in foodstuffs and biological specimens: a review. AB - Selenium is an essential element for human health. It has been recognized as an antioxidant and chemopreventive agent in cancer. Selenium is known to develop its biological activity via selenocysteine residue in the catalytically active centre of selenoproteins. The main source of selenium in human beings is the diet. However, in several regions of the world the content of selenium in diet has been estimated insufficient for a correct expression of the proteins. The beneficial effects of selenium on human health are strongly dependent on its chemical form and concentration. This review critically evaluated the state-of-the art of selenium speciation in biological matrices mainly focused in nutritional and food products. Besides the number of publications related to selenium speciation, isolation and accurate characterization and quantification of selenium species is still a challenge. Hyphenated techniques based on coupling chromatography separation with inductively coupled plasma spectrometry (ICP-MS) and its combination with molecular mass spectrometry (ESI-MS, ESI-MS-MS and MALDI-TOF) and isotopic dilution allow identification, quantification and structural characterization of selenium species. Particular attention is paid in the development of Se-enriched food and nutritional products and how the application of the techniques mentioned above is mandatory to get reliable results on selenium metabolisms in these particular matrices. PMID- 19185113 TI - Determination of trace amounts of sodium and lithium in zirconium dioxide (ZrO2) using liquid electrode plasma optical emission spectrometry. AB - This paper describes a quantitative measurement of trace elements (Na, Li) in high purity zirconium dioxide powder using liquid electrode plasma optical emission spectrometry (LEP-OES). Conventionally, for such type of measurements, inductively coupled plasma optical emission spectrometry (ICP-OES) is frequently employed. The detection limits of elements in zirconium by ICP-OES are degraded due to the spectra interference between the trace elements and zirconium of the matrix, because zirconium is a line rich element in spectra obtained by ICP-OES. LEP-OES is an elemental analysis method developed by the authors. The measurement principle is simple, as follows. Sample solution is put into a narrow channel on a small cuvette and voltage pulse is applied from both ends of the channel. At the center of the channel which is made narrower, the voltage and current are concentrated there, and plasma is generated. From the emission of the plasma, the quantitative analysis of the elements in the solution is achieved. The LEP-OES has the property that the emission of zirconium is relatively weak, so that highly sensitive measurement of trace elements in zirconium matrix can be conducted without interference. Sample solution is prepared by dissolving high purity zirconium dioxide powder and trace amounts of Na or Li with sulfuric acid. The voltage dependence and the pulse width dependence of optical emission spectra are also investigated. With increase of the voltage or the pulse width, the ratio of emission intensities of Na to those of hydrogen increases. This suggests that the ratio of sensitivity of two elements is variable, that means the element selectivity is controllable to some extent by the measurement conditions in LEP OES. In the case of Na and H, the ratio can be controlled from 7.4 to 21.6%. Finally, the detection limits (3S.D.) of the trace elements, Na and Li, in 4000 microg g(-1) zirconium dioxide aqueous solution are found to be 0.02 and 0.133 microg g(-1), respectively. These values correspond to 5 microg g(-1) for Na, 33.25 microg g(-1) for Li in original high purity zirconium dioxide powder. The correlation coefficient of calibration curve was 0.995 for Na, 0.985 for Li. Those are comparable to the literature values of detection limits using ICP-OES. PMID- 19185114 TI - Spectral quality assessment and application for gel-based matrix-assisted laser desorption ionization-time of flight tandem mass spectrometer. AB - Gel-based matrix-assisted laser desorption ionization-time of flight tandem mass spectrometer (MALDI TOF/TOF MS) is one of the dominant methods of current proteomics, utilizing both peptide mass fingerprinting (PMF) and peptide fragment fingerprinting (PFF) for protein identification on a spot-to-spot basis. However, the unique impact of the quality of the corresponding mass spectrometry spectra remains largely unreported, and has motivated the development and use of an automatic spectra-assessment method. In this study, a multi-variant regression approach has been utilized to assess spectral quality for both PMF and PFF spectra obtained from MALDI TOF/TOF MS. The assessment index has been applied to investigations of MASCOT search results. Systematic examination of two large scale sets of human liver tissue data has proved that spectral quality was a key factor in significant matching. Based on large-scale investigations on individual PMF search, individual PFF search and their combination, respectively, the filtering of bad quality spectra or spots proves to be an efficient way to improve search efficiency of all search modes in MASCOT. Meanwhile, a validation method based on score differences between normal and decoy (reverse or random) database searches is proposed to precisely define the positive matches. Further analysis showed that spectral quality assessment was also efficient in representing the quality of 2-DE gel spots and promoted the discovery of potential post-translation modifications. PMID- 19185115 TI - Determination of alpha-linolenic acid and linoleic acid in edible oils using near infrared spectroscopy improved by wavelet transform and uninformative variable elimination. AB - This paper proposes an analytical method for simultaneous near-infrared (NIR) spectrometric determination of alpha-linolenic and linoleic acid in eight types of edible vegetable oils and their blending. For this purpose, a combination of spectral wavelength selection by wavelet transform (WT) and elimination of uninformative variables (UVE) was proposed to obtain simple partial least square (PLS) models based on a small subset of wavelengths. WT was firstly utilized to compress full NIR spectra which contain 1413 redundant variables, and 42 wavelet approximate coefficients were obtained. UVE was then carried out to further select the informative variables. Finally, 27 and 19 wavelet approximate coefficients were selected by UVE for alpha-linolenic and linoleic acid, respectively. The selected variables were used as inputs of PLS model. Due to original spectra were compressed, and irrelevant variables were eliminated, more parsimonious and efficient model based on WT-UVE was obtained compared with the conventional PLS model with full spectra data. The coefficient of determination (r(2)) and root mean square error prediction set (RMSEP) for prediction set were 0.9345 and 0.0123 for alpha-linolenic acid prediction by WT-UVE-PLS model. The r(2) and RMSEP were 0.9054, 0.0437 for linoleic acid prediction. The good performance showed a potential application using WT-UVE to select NIR effective variables. WT-UVE can both speed up the calculation and improve the predicted results. The results indicated that it was feasible to fast determine alpha linolenic acid and linoleic acid content in edible oils using NIR spectroscopy. PMID- 19185116 TI - Prediction of sensory properties of Brazilian Arabica roasted coffees by headspace solid phase microextraction-gas chromatography and partial least squares. AB - Volatile compounds in fifty-eight Arabica roasted coffee samples from Brazil were analyzed by SPME-GC-FID and SPME-GC-MS, and the results were compared with those from sensory evaluation. The main purpose was to investigate the relationships between the volatile compounds from roasted coffees and certain sensory attributes, including body, flavor, cleanliness and overall quality. Calibration models for each sensory attribute based on chromatographic profiles were developed by using partial least squares (PLS) regression. Discrimination of samples with different overall qualities was done by using partial least squares discriminant analysis (PLS-DA). The alignment of chromatograms was performed by the correlation optimized warping (COW) algorithm. Selection of peaks for each regression model was performed by applying the ordered predictors selection (OPS) algorithm in order to take into account only significant compounds. The results provided by the calibration models are promising and demonstrate the feasibility of using this methodology in on-line or routine applications to predict the sensory quality of unknown Brazilian Arabica coffee samples. According to the PLS DA on chromatographic profiles of different quality samples, compounds 3 methypropanal, 2-methylfuran, furfural, furfuryl formate, 5-methyl-2 furancarboxyaldehyde, 4-ethylguaiacol, 3-methylthiophene, 2-furanmethanol acetate, 2-ethyl-3,6-dimethylpyrazine, 1-(2-furanyl)-2-butanone and three others not identified compounds can be considered as possible markers for the coffee beverage overall quality. PMID- 19185117 TI - Spectrophotometric determination of acidity and tautomeric constants and hydrogen bonding strength for a new Schiff base using hard modeling and multivariate curve resolution alternative least squares methods. AB - The equilibria of a new Schiff base derived from 3,6 bis((aminoethyl)thio)pyridazine were studied spectrophotometrically with the aid of factor-analytical methods. Hard modeling program was used for determination of the acidity constants of the Schiff base in dimethylformamide (DMF)/water mixture (30:70 v/v). In this method acidity constant equations act as hard models and the score vectors obtained by decomposing of absorbance data matrix will be linear combinations of equilibrium concentrations of species that exist in the absorption matrix. Two rank annihilation factor analysis (TRAFA) was used as a standard method to investigate the accuracy of the method. The tautomerization constant, K(t), of the Schiff base solution in various DMF/water mixtures has also been determined using spectral variations of the Schiff base solutions in various volume ratios of water with the aid of evolving factor analysis (EFA) and multivariate curve resolution alternative least squares (MCR-ALS) methods. In addition the intramolecular hydrogen bonding strength and its related thermodynamic parameters have been determined using MCR-ALS and spectral variation of the Schiff base solutions in different temperatures. PMID- 19185118 TI - In situ solvent formation microextraction based on ionic liquids: a novel sample preparation technique for determination of inorganic species in saline solutions. AB - In this research, a novel microextraction technique based on ionic liquids (ILs) termed in situ solvent formation microextraction (ISFME) is developed. In this method, small amount of sodium hexafluorophosphate (NaPF(6), as an ion-pairing agent) was added to the sample solution containing very small amount of 1-hexyl-3 methylimidazolium tetrafluoroborate ([Hmim][BF(4)], as hydrophilic IL). A cloudy solution was formed as a result of formation of fine droplets of 1-hexyl-3 methylimidazolium hexafluorophosphate [Hmim][PF(6)]. After centrifuging, the fine droplets of the extractant phase settled to the bottom of the conical-bottom glass centrifuge tube. ISFME is a simple and rapid method for extraction and preconcentration of metal ions from water samples and can be applied for the sample solutions containing very high concentrations of salt. Furthermore, this technique is much safer in comparison with the organic solvent extraction. Reliability of the introduced methodology was evaluated by analyzing water reference material. ISFME was successfully applied to determining mercury (II) in several real water samples. Michler thioketone (TMK) was chosen as a complexing agent. Analysis was carried out using spectrophotometric detection method. Type and amount of IL, temperature and the other parameters were optimized. Under the optimum conditions, the limit of detection (LOD) was 0.7 ng mL(-1) and the relative standard deviation (R.S.D.) was 1.94% for 40 ng mL(-1) mercury. PMID- 19185119 TI - Speciation analysis of inorganic arsenic in natural water by carbon nanofibers separation and inductively coupled plasma mass spectrometry determination. AB - In this paper, carbon nanofibers (CNFs) as a novel solid phase extraction sorbent were developed for speciation preconcentration and separation of inorganic arsenic species As(III) and As(V) prior to determination by inductively coupled plasma mass spectrometry (ICP-MS). It was found that during all the steps of the separation, As(III) was selectively sorbed on the microcolumn packed with CNFs within a pH range of 1.0-3.0 in the presence of ammonium pyrroinedithiocarbamate (APDC), while As(V) was passed through the microcolumn without the retention. Various experimental parameters affecting the separation and determination of As(III) and As(V) have been investigated in detail. Under the optimized conditions, the detection limits of this method for As(III) were 0.0045 ng mL(-1) with an enrichment factor of 33 and 0.24 ng mL(-1) for As(V), and the relative standard deviations for As(III) and As(V) were 2.6% and 1.9% (n=9, c=1.0 ng mL( 1)), respectively. In order to verify the accuracy of the method, a certified reference of water sample was analyzed, and the results obtained were in good agreement with the certified values. The proposed method was applied for the analysis of inorganic arsenic species in groundwater and lake water with the recovery of 92-106%. PMID- 19185120 TI - Accelerated solvent-based extraction and enrichment of selected plasticisers and 4-nonylphenol, and extraction of tin from organotin sources in sediments, sludges and leachate soils. AB - Enrichment techniques have become an important feature in the trace analysis of oestrogen mimicking chemicals in the environment. Recent developments such as accelerated solvent extraction (ASE) have improved extraction recoveries in a wide variety of solid matrices including sediments, sludges and leachate soils. Such samples taken from the Irish Midlands Shannon Catchment region during the winter of 2004/5 and suspected to contain certain xenooestrogens or hormonally active agents were extracted using this technique, which was then coupled with high performance liquid chromatography (HPLC) for quantification purposes. ASE was thus employed to both isolate and pre-concentrate targeted analytes using the minimum amount of solvent hence making extractions more conservational. Two simple, yet extremely sensitive liquid chromatographic methods were developed based on UV detection; one for phthalates and one for alkylphenols, with recoveries reaching up to 92.0%. Acid digestion was used for the extraction of the tin and organotin compounds with analysis by polarography. In river sediment, levels of up to 24.4 mg kg(-1) phthalate, 1.14 mg kg(-1) 4-nonylphenol and 118 mg kg(-1) tin were found. In leachate sediments, values up to 49.8 mg kg(-1) phthalate, 1.57 mg kg(-1) 4-nonylphenol, and 36.0 mg kg(-1) tin were determined. In sludge, values up to 174 mg kg(-1) phthalate and 22.8 mg kg(-1) 4-nonylphenol were quantified. The highest value of tin (118 mg kg(-1)) was found present in an area of high leisure craft activity. Typical sediment levels of tin at other river locations ranged between 1.20 and 37.5 mg kg(-1). PMID- 19185121 TI - Microwave-assisted double insert vapour-phase digestion of organic samples. AB - A microwave-assisted double insert multimode vapour-phase digestion method was developed for the digestion of organic samples. The experimental set-up was based on a third generation-type teflon microwave vessel, equipped with an automatic pressure regulating type vessel cover. A borosilicate glass holder insert, containing a smaller quartz sample insert, was fitted inside the vessel. Sulphuric acid was added to the holder insert as a microwave absorbing and temperature transferring liquid, which transferred heat to the sample insert (into which the sample was weighed) and charred the sample material. Oxidation of the sample material was carried out simultaneously with charring using nitric acid vapour, which was generated by the 1:1 (v/v) sulphuric acid-nitric acid mixture located in the bottom of the microwave vessel. This set-up generated high digestion efficiency, without any of the interferences normally associated with direct sulphuric acid usage. The method was used for determining the concentrations of Cd, Cr, Cu, Mn, Mo, Zn and Fe in certified organic reference materials using ICP-OES instrumentation. The certified organic reference materials were NRCC DOLT-2 dogfish liver, NIST-SRM 1577b bovine liver and IRMM VDA cadmium in polyethylene No. 001 and No. 004. The results were in good agreement with the certified values, forepart from Cd. For Cd the results were lower than the certified values due to volatilization losses. Sample materials that could not be digested by an earlier procedure were completely digested during a single-step, 30 min digestion. The tested sample materials included certified reference materials, 3-nitrobenzoic acid (3-NBA) and pike (Esox lucius) muscle. The residual carbon concentrations in the digestion solutions were below the detection limit of the TOC instrument. This type of digestion method is described here for the first time in the literature. PMID- 19185122 TI - An interior needle electropolymerized pyrrole-based coating for headspace solid phase dynamic extraction. AB - A headspace solid-phase dynamic extraction (HS-SPDE) technique was developed by the use of polypyrrole (PPy) sorbent, electropolymerized inside the surface of a needle, as a possible alternative to solid-phase microextraction (SPME). Thermal desorption was subsequently, employed to transfer the extracted analytes into the injection port of a gas chromatography-mass spectrometry (GC-MS). The PPy sorbent including polypyrrole-dodecyl sulfate (PPy-DS) was deposited on the interior surface of a stainless steel needle from the corresponding aqueous electrolyte by applying a constant deposition potential. The homogeneity and the porous surface structure of the coating were examined using the scanning electron microscopy (SEM). The developed method was applied to the trace level extraction of some polycyclic aromatic hydrocarbons (PAHs) from aqueous sample. In order to enhance the extraction efficiency and increase the partition coefficient of analytes, the stainless steel needle was cooled at 5 degrees C, while the sample solution was kept at 80 degrees C. Optimization of influential experimental conditions including the voltage of power supply, the time of PPy electrodeposition, the extraction temperature, the ionic strength and the extraction time were also investigated. The detection limits of the method under optimized conditions were in the range of 0.002-0.01 ng mL(-1). The relative standard deviations (R.S.D.) at a concentration level of 0.1 ng mL(-1) were obtained between 7.54 and 11.4% (n=6). The calibration curves of PAHs showed linearity in the range of 0.01-10 ng mL(-1). The proposed method was successfully applied to the extraction of some selected PAHs from real-life water samples and the relative recoveries were higher than 90% for all the analytes. PMID- 19185123 TI - On-line coupling of solid-phase extraction to liquid chromatography-tandem mass spectrometry for the determination of macrolide antibiotics in environmental water. AB - An automated on-line solid-phase extraction-liquid chromatography-tandem mass spectrometry (SPE-LC-MS/MS) system was developed for the determination of macrolide antibiotics including erythromycin (ETM), roxithromycin (RTM), tylosin (TLS) and tilmicosin (TMC) in environmental water samples. A Capcell Pak MF Ph-1 column packed with restricted access material (RAM) was used as SPE column for the concentration of the analytes and clean-up of the sample. One milliliter water sample was injected into the conditioned SPE column and the matrix was washed out with 3 mL high purity water. By rotation of the switching valve, macrolides (MLs) were eluted in the back-flush mode and transferred to the analytical column by the chromatographic mobile phase. The matrix effect was evaluated by the directly injection LC-MS and on-line SPE-LC-MS methods. The limits of detection (LODs) and limits of quantification (LOQs) obtained are in the range of 2-6 and 7-20 ng L(-1), respectively, which means that the proposed method is suitable for trace analysis of MLs at low level concentration. The intra- and inter-day precisions are in the range of 2.9-7.2% and 3.3-8.9%, respectively. In the three fortified levels (20, 200 and 2000 ng L(-1)), recoveries of MLs ranging from 86.5% to 98.3% are obtained. PMID- 19185124 TI - Evaluation of tris(4,7-diphenyl-1,10-phenanthrolinedisulfonate)ruthenium(II) as a chemiluminescence reagent. AB - Previous studies have suggested that tris(4,7-diphenyl-1,10 phenanthrolinedisulfonate)ruthenium(II) (Ru(BPS)(3)(4-)) has great potential as a chemiluminescence reagent in acidic aqueous solution. We have evaluated four different samples of this reagent (two commercially available and two synthesised in our laboratory) in comparison with tris(2,2'-bipyridine)ruthenium(II) (Ru(bipy)(3)(2+)) and tris(1,10-phenanthroline)ruthenium(II) (Ru(phen)(3)(2+)), using a range of structurally diverse analytes. In general, Ru(BPS)(3)(4-) produced more intense chemiluminescence, but the oxidised Ru(BPS)(3)(3-) species is less stable in aqueous solution than Ru(bipy)(3)(3+) and produced a greater blank signal than Ru(bipy)(3)(3+) or Ru(phen)(3)(3+), which had a detrimental effect on sensitivity. Although the complex is often depicted with the sulfonate groups of the BPS ligand in the para position on the phenyl rings, NMR characterisation revealed that the commercially available BPS material used in this study was predominantly the meta isomer. PMID- 19185125 TI - An ultra-clean technique for accurately analysing Pb isotopes and heavy metals at high spatial resolution in ice cores with sub-pg g(-1) Pb concentrations. AB - Measurements of Pb isotope ratios in ice containing sub-pg g(-1) concentrations are easily compromised by contamination, particularly where limited sample is available. Improved techniques are essential if Antarctic ice cores are to be analysed with sufficient spatial resolution to reveal seasonal variations due to climate. This was achieved here by using stainless steel chisels and saws and strict protocols in an ultra-clean cold room to decontaminate and section ice cores. Artificial ice cores, prepared from high purity water were used to develop and refine the procedures and quantify blanks. Ba and In, two other important elements present at pg g(-1) and fg g(-1) concentrations in Polar ice, were also measured. The final blank amounted to 0.2+/-0.2 pg of Pb with (206)Pb/(207)Pb and (208)Pb/(207)Pb ratios of 1.16+/-0.12 and 2.35+/-0.16, respectively, 1.5+/-0.4 pg of Ba and 0.6+/-2.0 fg of In, most of which probably originates from abrasion of the steel saws by the ice. The procedure was demonstrated on a Holocene Antarctic ice core section and was shown to contribute blanks of only approximately 5%, approximately 14% and approximately 0.8% to monthly resolved samples with respective Pb, Ba and In concentrations of 0.12 pg g(-1), 0.3 pg g(-1) and 2.3 fg g(-1). Uncertainties in the Pb isotopic ratio measurements were degraded by only approximately 0.2%. PMID- 19185126 TI - Development of a certified reference material for the content of nitroimidazole parent drugs and hydroxy metabolites in pork meat. AB - Nitroimidazoles have been applied in the past to poultry and pigs to treat protozoan diseases and to combat bacterial infections, but due to adverse health effects their use in food-producing animals has meanwhile been banned in the EU. The request for a certified reference material in a representative matrix was stipulated by the responsible Community Reference Laboratory and is underpinned by the need to improve the accuracy and comparability of measurement data and to establish metrological traceability of analytical results. The Institute for Reference Materials and Measurements (IRMM) has responded to this demand by developing and producing a new certified matrix reference material, ERM-BB124. This incurred lyophilised pork meat material was certified according to ISO guides 34 and 35 for the mass fractions of six nitroimidazole compounds. Processing of the frozen muscle tissue to the final material was accomplished by application of cutting, freeze-drying, mixing and milling techniques. Homogeneity and stability measurements were performed using liquid chromatography tandem mass spectrometry. The relative standard uncertainty due to possible heterogeneity showed to be below 1.8% for all analytes. Potential degradation during transport and storage was assessed by isochronous stability studies. No significant instability was detected at a storage temperature of -20 degrees C for a shelf life of 2 years. The certified mass fraction values were assigned upon evaluation of the data acquired in an international laboratory inter-comparison involving 12 expert laboratories using different sample preparation procedures, but exclusively LC-MS/MS methods. Relative standard uncertainty contributions for the characterisation (between-lab variation of mean values) were found to be between 1.6 and 4.8%. Certified values for five analytes were in the range of 0.7 to 6.2 microg kg(-1), with expanded relative uncertainties ranging between 7 and 14%. Dimetridazole could be certified as "<0.25 microg kg(-1) with a probability of 95%". All values are traceable to the International System of Units (SI). The material is intended to be used for method validation purposes (including trueness estimation) and for method performance assessment. PMID- 19185127 TI - Langmuir-Blodgett film based on MEH-PPV for cholesterol biosensor. AB - Cholesterol oxidase (ChOx) has been immobilized onto conducting poly[2-methoxy,5 (2'-ethyl-hexyloxy)-1,4-phenylene vinylene] (MEH-PPV)/stearic acid (SA) Langmuir Blodgett film transferred onto octadecanethiol (ODT) modified gold plate. The ChOx/MEH-PPV/SA LB film bioelectrode exhibits has been characterized by FT-IR, contact angle, and atomic force microscopy. The response of the ChOx/MEH-PPV/SA LB film bioelectrode carried out using differential pulse voltammetry (DPV) studies reveal linearity from 1.29 to 12.91 mM of cholesterol concentration and response time as 30 s. This ChOx/MEH-PPV/SA bioelectrode exhibits values of correlation coefficient as 0.9939, standard deviation as 0.0029 microA and limit of detection as 1.66 mM. UV-visible spectrophotometer studies reveal that 5.2x10( 3) U of ChOx are actively working per cm(2) area of ChOx/MEH-PPV/SA LB film bioelectrode and this bioelectrode is thermally stable upto 55 degrees C with reusability of about 60 times. PMID- 19185128 TI - Specific detection of oxytetracycline using DNA aptamer-immobilized interdigitated array electrode chip. AB - An electrochemical sensing system for oxytetracycline (OTC) detection was developed using ssDNA aptamer immobilized on gold interdigitated array (IDA) electrode chip. A highly specific ssDNA aptamer that bind to OTC with high affinity was employed to discriminate other tetracyclines (TCs), such as doxycycline (DOX) and tetracycline (TET). The immobilized thiol-modified aptamer on gold electrode chip served as a biorecognition element for the target molecules and the electrochemical signals generated from interactions between the aptamers and the target molecules was evaluated by cyclic voltammetry (CV) and square wave voltammetry (SWV). The current decrease due to the interference of bound OTC, DOX or TET was analyzed with the electron flow produced by a redox reaction between ferro- and ferricyanide. The specificity of developed EC biosensor for OTC was highly distinguishable from the structurally similar antibiotics (DOX and TET). The dynamic range was determined to be 1-100 nM of OTC concentration in semi-logarithmic coordinates. PMID- 19185129 TI - A capacitive immunosensor for detection of cholera toxin. AB - Contamination of food with biological toxins as well as their potential use as weapons of mass destruction has created an urge for rapid and cost effective analytical techniques capable of detecting trace amounts of these toxins. This paper describes the development of a sensitive method for detection of cholera toxin (CT) using a flow-injection capacitive immunosensor based on self-assembled monolayers. The sensing surface consists of monoclonal antibodies against the B subunit of CT (anti-CT), immobilized on a gold transducer. Experimental results show that the immunosensor responded linearly to CT concentrations in the range from 1.0x10(-13) to 1.0x10(-10) M under optimized conditions. The limit of detection (LOD) was 1.0x10(-14) M. Two more analytical methods were employed for detection of CT using the same antibody namely, sandwich ELISA and surface plasmon resonance (SPR)-based immunosensor. The former had an LOD of 1.2x10(-12) M and a working range from 3.7x10(-11) to 2.9x10(-10) M whereas, the later had an LOD of 1.0x10(-11) M and a linearity ranging from 1.0x10(-9) to 1.0x10(-6) M. These results demonstrate that the developed capacitive immunosensor system has a higher sensitivity than the other two techniques. The binding affinity of CT to the immobilized anti-CT was determined using the SPR-based immunosensor and an association constant (K(A)) of 1.4x10(9) M(-1) was estimated. PMID- 19185130 TI - A novel ratiometric fluorescent Fe3+ sensor based on a phenanthroimidazole chromophore. AB - Phenanthroimidazole derivative 1 has been developed as a rare example of ratiometric fluorescent sensors for Fe(3+). Interestingly, upon treatment with Fe(3+), the sensor displayed a ratiometric fluorescent response with an enhancement of the ratios of emission intensities at 440 and 500 nm from 0.36 to 3.24. The detection range of the sensor for Fe(3+) is in the 1.0x10(-5)-1.5x10( 4) M concentration range and the detection limit is 5.26x10(-6) M. In addition, the sensor showed good selectivity to Fe(3+) with the selectivity coefficients (K(Fe3+) = S(Fe3+)/S(0) of Fe(3+) over other metal ions tested in the range of 5 68. PMID- 19185131 TI - New rapid methods for determination of total LAS in sewage sludge by high performance liquid chromatography (HPLC) and capillary electrophoresis (CE). AB - Linear alkylbenzene sulfonates (LAS) are the most common synthetic anionic surfactant used in domestic and industrial detergents, with a global production of 2.4x10(6) t year(-1). After use and disposal, LAS may enter the environment by one of the several routes, including by direct discharge to surface water or discharge to water from sewage treatment plants. Sewage treatment plants break down LAS only partly: some of them remain in effluent and other fraction is adsorbed in sewage solid. New and rapid methods for determination of total LAS from sewage sludge based on microwave assisted extraction and HPLC-FL and CE-DAD determination are proposed. The extraction of total LAS is carried out by using microwaves energy, an extraction time of 10 min and 5 mL of methanol. For HPLC-FL determination, mobile phase acetonitrile-water was used, comprising 60% (v/v) from 0 to 1 min and a flow rate of 1 mL min(-1) programmed to 100% acetonitrile between 1 and 2 min and a flow rate of 2 mL min(-1). The final composition was maintained for a further 5 min. The determination of total LAS by CE-DAD was performed in a phosphate buffer (10 mM, pH 9). The separation voltage was 25 kV and the temperature of the capillary was 30 degrees C. Injections were performed in the pressure mode and the injection time was set at 12 s. The determination of total LAS is carried out in less than 5 min. The methods did not require clean-up or preconcentration steps. Detection limit for total LAS in the sludge was 3.03 mg kg(-1) using HPLC-FL and 21.0 mg kg(-1) using CE-DAD, and recoveries were >85% using both determination methods. Concentrations of total LAS obtained using both methods were compared with the sum of concentrations of homologues LAS C-10, LAS C-11, LAS C-12 and LAS C-13 obtained using microwaves assisted extraction and HPLC-FL and CE-DAD determination. PMID- 19185132 TI - Authentication and quantitative analysis on the chemical profile of Xanthium fruit (Cang-Er-Zi) by high-performance liquid chromatography-diode-array detection tandem mass spectrometry method. AB - A high-performance liquid chromatographic method using diode-array detection and electrospray ionization tandem mass spectrometry (HPLC-DAD-ESI-MS/MS) was developed for the qualitative and quantitative analysis on Xanthium fruit, a commonly used traditional Chinese medicine. In this study, 7 characteristic components, 1-O-caffeoylquinic acid, 4-O-caffeoylquinic acid, chlorogenic acid, 1,5-O-di-caffeoylquinic acid, 1,3-O-di-caffeoylquinic acid, 4,5-O-di caffeoylquinic acid and 1,3,5-O-tri-caffeoylquinic acid were identified and quantified by a validated HPLC-DAD method, and a fingerprint comprised of 12 markers was established under the same operating conditions. Furthermore, HPLC ESI-MS/MS method was successfully used to deduce the structure of three main constituents. On the basis of the established chromatographic profiles, 30 populations of cocklebur samples including 3 related species and 1 unknown species were divided into 3 chemotypes, indicated that place of origin significantly influences the kinds and content of components in cocklebur, and hence affects their quality. The simultaneous determination of 7 caffeoylquinic acids in the 30 samples showed a great variety in the amounts of caffeoylquinic acids present. The study indicated that some species such as Xanthium mongolicum of the genus Xanthium might be suitable for development as new alternative sources of caffeoylquinic acids to supplement the officially listed Xanthium species, and the abundant constituents such as chlorogenic acid perhaps should be recorded in some authorized publications and applied to the quality control or quality evaluation for Xanthium in China. The entire analytical procedure is reproducible and suitable for the authentication and quantification of Xanthium fruits. PMID- 19185133 TI - Spectrum-effect relationships between ultra performance liquid chromatography fingerprints and anti-bacterial activities of Rhizoma coptidis. AB - The fingerprints of Rhizoma coptidis from various sources were established by ultra performance liquid chromatography (UPLC) and the anti-bacterial activities of R. coptidis on Escherichia coli (E. coli) growth was studied by microcalarimetry. The UPLC fingerprints were evaluated using similarity analysis (SA) and hierarchical clustering analysis (HCA). Some quantitative parameters obtained from the thermo-genic curves of E. coli growth affected by R. coptidis were analyzed using principal component analysis (PCA). The spectrum-effect relationships between UPLC fingerprints and anti-bacterial activities were investigated using canonical correlation analysis (CCA). The results showed that close correlation existed between the spectrum-effect relationships. Berberine, jateorrhizine and palmatine in the UPLC fingerprints might be the main anti bacterial components. The anti-bacterial activities of R. coptidis were related with the main active constituents, along with the production place and the harvesting time of this herb, the latitude and longitude of the place. This work provides a general model of the combination of UPLC and microcalorimetry to study the spectrum-effect relationships of R. coptidis, which can be used to discover principle components of it on bioactivity. PMID- 19185134 TI - Mitral valve surgery for functional mitral regurgitation: should moderate to severe tricuspid regurgitation be treated? A propensity score analysis. PMID- 19185135 TI - Norman E. Shumway, MD, PhD: visionary, innovator, humorist. PMID- 19185136 TI - POINT: Prosthesis-patient mismatch does not affect survival for patients greater than 70 years of age undergoing bioprosthetic aortic valve replacement. AB - OBJECTIVE: The purpose of this investigation was to examine the impact of prosthesis-patient mismatch after bioprosthetic aortic valve replacement on long term survival in patients greater than 70 years of age compared with those less than or equal to 70 years of age. METHODS: Between 1992 and 2007, 1399 patients underwent bioprosthetic aortic valve replacement, including 881 (63%) patients older than 70 years of age. Prosthesis-patient mismatch was defined as severe (prosthetic effective orifice area/body surface area < 0.65 cm(2)/m(2)), moderate (0.65-0.85 cm(2)/m(2)), or absent (>0.85 cm(2)/m(2)). For patients less than or equal to 70 years of age, mismatch was severe in 62 (12%), moderate in 251 (48%), and absent in 205 (40%). For patients greater than 70 years of age, mismatch was severe in 109 (12%), moderate in 451 (51%), and absent in 321 (37%). There was no difference in the distribution of prosthesis-patient mismatch between age groups (P = .50). RESULTS: For patients less than or equal to 70 years, prosthesis patient mismatch was associated with impaired long-term survival (P = .02). Survival at 5 and 10 years was 61% +/- 7% and 28% +/- 12% for severe mismatch, 65% +/- 3% and 40% +/- 5% for moderate mismatch, and 73% +/- 5% and 46% +/- 9% for no mismatch. For patients greater than 70 years, prosthesis-patient mismatch did not affect long-term survival (P = .25). Survival at 5 and 10 years was 62% +/- 5% and 42% +/- 6% for severe mismatch, 62% +/- 2% and 30% +/- 5% for moderate mismatch, and 53% +/- 4% and 29% +/- 5% for absent mismatch. CONCLUSIONS: After bioprosthetic aortic valve replacement, prosthesis-patient mismatch had a negative impact on late survival for patients less than or equal to 70 years of age, but for patients greater than 70 years of age, prosthesis-patient mismatch did not influence late survival. PMID- 19185137 TI - COUNTERPOINT: Aortic valve replacement: size does matter. PMID- 19185138 TI - Repair-oriented classification of aortic insufficiency: impact on surgical techniques and clinical outcomes. AB - OBJECTIVE: Valve repair for aortic insufficiency requires a tailored surgical approach determined by the leaflet and aortic disease. Over the past decade, we have developed a functional classification of AI, which guides repair strategy and can predict outcome. In this study, we analyze our experience with a systematic approach to aortic valve repair. METHODS: From 1996 to 2007, 264 patients underwent elective aortic valve repair for aortic insufficiency (mean age - 54 +/- 16 years; 79% male). AV was tricuspid in 171 patients bicuspid in 90 and quadricuspid in 3. One hundred fifty three patients had type I dysfunction (aortic dilatation), 134 had type II (cusp prolapse), and 40 had type III (restrictive). Thirty six percent (96/264) of the patients had more than one identified mechanism. RESULTS: In-hospital mortality was 1.1% (3/264). Six patients experienced early repair failure; 3 underwent re-repair. Functional classification predicted the necessary repair techniques in 82-100% of patients, with adjunctive techniques being employed in up to 35% of patients. Mid-term follow up (median [interquartile range]: 47 [29-73] months) revealed a late mortality rate of 4.2% (11/261, 10 cardiac). Five year overall survival was 95 +/ 3%. Ten patients underwent aortic valve reoperation (1 re-repair). Freedoms from recurrent Al (>2+) and from AV reoperation at 5 years was 88 +/- 3% and 92 +/- 4% respectively and patients with type I (82 +/- 9%; 93 +/- 5%) or II (95 +/- 5%; 94 +/- 6%) had better outcomes compared to type III (76 +/- 17%; 84 +/- 13%). CONCLUSION: Aortic valve repair is an acceptable therapeutic option for patients with aortic insufficiency. This functional classification allows a systematic approach to the repair of Al and can help to predict the surgical techniques required as well as the durability of repair. Restrictive cusp motion (type III), due to fibrosis or calcification, is an important predictor for recurrent Al following AV repair. PMID- 19185140 TI - Effects of on- and off-pump coronary artery surgery on graft patency, survival, and health-related quality of life: long-term follow-up of 2 randomized controlled trials. AB - OBJECTIVE: Off-pump coronary artery bypass grafting reduces postoperative morbidity and uses fewer resources than conventional surgical intervention with cardiopulmonary bypass. However, only 15% to 20% of coronary artery bypass grafting operations use off-pump coronary artery bypass. One reason for not using off-pump coronary artery bypass might be the surgeon's concern about the long term patency of grafts performed with this technique. Therefore our objective was to compare long-term outcomes in patients randomized to off-pump coronary artery bypass or coronary artery bypass grafting with cardiopulmonary bypass. METHODS: Participants in 2 randomized trials comparing off-pump coronary artery bypass and coronary artery bypass grafting with cardiopulmonary bypass were followed up for 6 to 8 years after surgical intervention to assess graft patency, major adverse cardiac-related events, and health-related quality of life. Patency was assessed by using multidetector computed tomographic coronary angiographic analysis with a 16-slice scanner. Two blinded observers classified proximal, body, and distal segments of each graft as occluded or not. Major adverse cardiac-related events and health-related quality of life were obtained from questionnaires given to participants and family practitioners. RESULTS: Patency was studied in 199 and health-related quality of life was studied in 299 of 349 survivors. There was no evidence of attrition bias. The likelihood of graft occlusion was no different between off-pump coronary artery bypass (10.6%) and coronary artery bypass grafting with cardiopulmonary bypass (11.0%) groups (odds ratio, 1.00; 95% confidence interval, 0.55-1.81; P > .99). Graft occlusion was more likely at the distal than the proximal anastomosis (odds ratio, 1.11; 95% confidence interval, 1.02-1.20). There were also no differences between the off-pump coronary artery bypass and coronary artery bypass grafting with cardiopulmonary bypass groups in the hazard of death (hazard ratio, 1.24; 95% confidence interval, 0.72-2.15) or major adverse cardiac-related events or death (hazard ratio, 0.84; 95% confidence interval, 0.58-1.24), or mean health-related quality of life across a range of domains and instruments. CONCLUSIONS: Long-term health outcomes with off-pump coronary artery bypass are similar to those with coronary artery bypass grafting with cardiopulmonary bypass when both operations are performed by experienced surgeons. PMID- 19185142 TI - Is preoperative serum creatinine a reliable indicator of outcome in patients undergoing coronary artery bypass surgery? AB - OBJECTIVE: Evaluating renal function by calculating creatinine clearance as an alternative measure to serum creatinine may give a better estimation of postoperative renal function in patients undergoing coronary artery bypass grafting. METHODS: Using our database, we conducted a retrospective review of the records of all 11,884 patients aged 21 years or older undergoing pure bypass grafting who required cardiopulmonary bypass. Preoperative renal function was categorized as normal renal function (serum creatinine 60 mL/min), occult renal insufficiency (serum creatinine 1.5 mg/dL and creatinine clearance 7 days) was higher in patients with occult renal insufficiency than in the normal group in univariable analysis. Multivariable logistic regression analysis demonstrated that patients with occult renal insufficiency compared with the group with normal renal function were at higher risk for mortality (odds ratio = 2.59, 95% confidence interval 1.15-5.86; P = .022) and prolonged hospital stay (>7 d) (odds ratio = 1.30, 95% confidence interval 1.08-1.57; P = .005). CONCLUSIONS: To identify higher-risk patients requiring special intensive care, and in whom new interventions can be performed to improve outcome, we recommend the preoperative calculation of creatinine clearance, especially in older women with a lower body mass index. PMID- 19185143 TI - Impact of preoperative clopidogrel in off pump coronary artery bypass surgery: a propensity score analysis. AB - OBJECTIVE: The aim of our study was to evaluate the impact of recent clopidrogel use before off-pump coronary artery bypass grafting on the postoperative risk of bleeding. METHODS: During the period January 2003 to December 2006, 1104 consecutive patients underwent off-pump coronary artery bypass grafting. Patients were divided into two groups according to the recent use of clopidrogel (within 7 days). We performed a propensity score to further adjust for differences between the patients with and without recent use of clopidrogel. RESULTS: Mean age was 64 +/- 14 years and 87% were male. The clopidrogel group had a greater incidence of patients in unstable condition, requiring emergency coronary bypass grafting, and with a high EuroSCORE. Propensity score analysis selected 88 patients with and 176 without recent use of clopidrogel. By propensity score, the clopidrogel group had higher requirements for fresh frozen plasma units (18.1% vs 8.5%; P = .02), reoperation owing to bleeding (5.6% vs 0.5%; P = .009), and higher need for postoperative mechanical ventilation (4% vs 10%; P = .04), whereas mortality and length of stay were similar between groups. CONCLUSION: Recent use of clopidogrel before off-pump coronary artery bypass grafting is associated with greater risk for bleeding with similar mortality rate. PMID- 19185144 TI - Factors associated with the development of aortic valve regurgitation over time after two different techniques of valve-sparing aortic root surgery. AB - OBJECTIVE: Early results after aortic valve-sparing root reconstruction are excellent. Longer-term follow-up, especially with regard to aortic valve function, is required for further judgment of these techniques. METHODS: Between July of 1993 and September of 2006, 108 consecutive patients (mean age 53.0 +/- 15.8 years) underwent the Yacoub operation (group Y) and 83 patients underwent the David operation (group D). Innovative multilevel hierarchic modeling methods were used to analyze aortic regurgitation over time. RESULTS: In general, aortic regurgitation increased with time in both groups. Factors associated with the development of a significant increase in aortic regurgitation were Marfan syndrome, concomitant cusp intervention, and preoperative aortic anulus dimension. In Marfan syndrome, the initial aortic regurgitation was higher in group Y versus group D (0.56 aortic regurgitation vs 0.29 aortic regurgitation, P = .049), whereas the mean annual progression rate of aortic regurgitation was marginally higher in group Y (0.132 aortic regurgitation vs 0.075 aortic regurgitation, P = .1). Concomitant cusp intervention was associated with a significant aortic regurgitation increase in both groups (P < .0001). There was a trend that smaller preoperative aortic annulus diameters in group D and larger diameters in group Y were associated with increased aortic regurgitation over time. CONCLUSION: In regard to aortic regurgitation grade over time, patients with Marfan syndrome and a large preoperative aortic annulus diameter were better treated with the reimplantation technique, whereas those with a smaller diameter were better treated with the remodeling technique. Concomitant free-edge plication of prolapsing cusps was disadvantageous in both groups. Considering these factors may serve to improve the aortic valve longevity after valve-sparing aortic root surgery. PMID- 19185145 TI - Management and decision-making strategy for systolic anterior motion after mitral valve repair. AB - OBJECTIVE: Systolic anterior motion can complicate mitral valve repair. It can have no clinical consequence or cause low cardiac output syndrome and hypotension. The management of systolic anterior motion in the operating room remains controversial: some groups advocate nonsurgical management, and others propose immediate surgical correction. Conventional hemodynamic measures require time and can be unsuccessful. While describing our experience, we propose a simple and innovative management and classification of this complication. METHODS: Presenting the data of 608 consecutive patients who underwent mitral valve repair for degenerative mitral valve disease, we describe a novel 2-step conservative management consisting of intravascular volume expansion and discontinuation of inotropic drug (step 1) and increasing afterload by means of ascending aortic manual compression while administering beta-blockers (step 2). We also describe a novel classification of systolic anterior motion: easy to revert (responding to step 1), difficult to revert (responding to step 2), or persistent. RESULTS: The overall incidence of systolic anterior motion was 9.8% (60/608): 40 patients had easy-to-revert systolic anterior motion, and 15 had difficult-to-revert systolic anterior motion. Five patients had a persistent condition and underwent surgical intervention within 48 hours. CONCLUSIONS: Systolic anterior motion after repair of a degenerative mitral valve is common. Surgical revision in the minority of patients unresponsive to standard conservative management is suggested. PMID- 19185146 TI - Predictors of recurrence and reoperation for prosthetic valve endocarditis after valve replacement surgery for native valve endocarditis. AB - OBJECTIVE: Surgical treatment of native valve endocarditis remains challenging, especially in cases with paravalvular destruction. Basic principles include complete debridement and reconstruction. This study is designed to evaluate the outcomes of surgical reconstruction of complex annular endocarditis using standard techniques and materials, including autologous and bovine pericardium. METHODS: From 1975 to 2000, 358 cases (357 patients, mean age 49 +/- 16 years, range 18-88 years) of native valve endocarditis were surgically managed. Bioprosthetic valves were implanted in 189 cases, and mechanical prostheses were implanted in 169 cases. A total of 78 cases of paravalvular destruction were identified: 62 annular abscesses, 8 fistulas, and 8 combined abscesses/fistulas. These were managed with 46 pericardial patches and 32 isolated suture reconstructions after radical debridement and prosthetic valve replacement. RESULTS: The overall early mortality was 8.4% (n = 30). The mortality with paravalvular destruction was 17.9%, and the mortality with simple leaflet infection was 5.7% (P = .001). The unadjusted survival at 20 years was 26.4% +/- 4.9% for bioprosthetic valves and 56.5% +/- 8.1% for mechanical prostheses (P = .007). The freedom from recurrent prosthetic valve endocarditis was 78.9% +/- 4.4% at 15 years. The freedom from reoperation for recurrent prosthetic valve endocarditis was 85.8% +/- 4.2% at 15 years. The freedom from reoperation after reconstruction for paravalvular destruction was 88.2% +/- 6.9% at 15 years. The freedom from mortality for recurrent prosthetic valve endocarditis was 92.7% +/- 3.4% at 15 years. The independent predictors of reoperation were age (hazard ratio 0.930, P = .005) and intravenous drug use/human immunodeficiency virus plus surgical technique (hazard ratio 12.8, P = .003 for patch reconstruction plus valve and hazard ratio 3.6, P = .038 for valve replacement only). Prosthesis type was not predictive when separated from intravenous drug use/human immunodeficiency virus (hazard ratio 3.268, P = .088). CONCLUSION: Paravalvular destruction is associated with a higher operative mortality. Native valve endocarditis can be managed with reasonable long-term survival and low rates of reinfection with radical debridement and pericardial reconstruction with bioprostheses and mechanical prostheses. The type of prosthesis implanted does not influence long-term outcome. Patients with a history of intravenous drug use and human immunodeficiency virus are at increased risk for recurrent infection and reoperation. PMID- 19185147 TI - Propensity analysis of survival after subcoronary or root replacement techniques for homograft aortic valve replacement. AB - OBJECTIVE: Homograft aortic valve replacement is associated with excellent clinical and hemodynamic outcomes. Valves are implanted predominantly by using 2 techniques: the freehand subcoronary technique or as an aortic root replacement. Our aim was to identify any difference in survival, durability, and clinical performance. METHODS: Demographic, operative, and clinical data were obtained retrospectively through case-note review. All operations were performed by a single surgeon. Propensity score-adjusted analysis was used by developing a nonparsimonious logistic regression model for implantation with subcoronary versus root replacement. Actuarial survival and freedom from valve-related events were compared with Kaplan-Meier curves and multivariable proportional hazard Cox regression. RESULTS: Between January 1, 1991, and January 1, 2001, 215 patients underwent aortic valve replacement with a homograft. The subcoronary technique was used in 131 (61%) patients. Eighty-four (39%) patients underwent free standing aortic root replacement. After propensity risk adjustment, the subcoronary implantation technique was associated with a decreased risk of 30-day death (adjusted odds ratio, 0.18; 95% confidence interval, 0.06-0.34; P = .03). Technique of insertion was not an independent predictor of overall mortality during follow-up after adjustment (propensity adjusted hazard ratio, 0.35; 95% confidence interval, 0.09-1.41; P = .18). There were no significant differences in 1- and 5-year actuarial survival, freedom from structural valve disease, endocarditis, or reoperation. CONCLUSIONS: Both the subcoronary and root replacement techniques for homograft aortic valve replacement are associated with excellent midterm survival and clinical performance. Root replacement was associated with an increased risk of perioperative death after adjustment for covariates by using propensity analysis. PMID- 19185148 TI - Noninvasive positive-pressure ventilation for extubation failure after cardiac surgery: Pilot safety evaluation. AB - OBJECTIVE: Extubation failure is a serious complication after cardiac surgery. The role of noninvasive positive-pressure ventilation for acute respiratory failure in patients undergoing cardiac surgery is unknown. This study aimed to assess the safety of implementing noninvasive positive-pressure ventilation in this setting and its impact on lung function and operative outcomes. METHODS: In a 6-month pilot prospective survey, the study population comprised 43 patients (32 were male with a mean age of 65.73 +/- 9 years; 3 heart transplantations, 18 coronary artery bypass grafts, 5 aortic dissections, and 17 valvular procedures; 34 active smokers, 25 with medically treated chronic obstructive pulmonary disease, 21 emergency/urgency procedures) who required noninvasive positive pressure ventilation for acute respiratory failure after initial weaning from a respirator. The cause of acute respiratory failure (classified as post cardiopulmonary bypass lung injury in 48.8% [21 patients], cardiogenic edema in 30.2% [13 patients], and pneumonia in 21% [9 patients]), length of noninvasive positive-pressure ventilation support, respiratory ratios (arterial oxygen tension/fraction of inspired oxygen assessed immediately before noninvasive positive-pressure ventilation, and every 6 hours after institution of pressure ventilation), and need for reintubation along with a set of predefined safety parameters were recorded. RESULTS: The mean length of noninvasive positive pressure ventilation support was 33.8 +/- 24.04 hours. Plotting respiratory ratios with length of noninvasive positive-pressure ventilation supports a significant improvement was already evident within the first 6-hour frame (133.6 +/- 39.5 vs 205 +/- 65.7; P < .001) for all causes. Noninvasive positive-pressure ventilation prevented intubation in 74.4% of the patients, with satisfactory recovery for post-cardiopulmonary bypass lung injury and cardiogenic dysfunction (90.5% and 69.2%, respectively) and poor results (55% reintubated) in those treated for pneumonia. Noninvasive positive-pressure ventilation safety approached 97.7%. CONCLUSION: In appropriate candidates, noninvasive positive pressure ventilation exerts favorable effects on lung function, preventing reintubation. The cost-effectiveness of its systematic use in this setting should be assessed. PMID- 19185149 TI - Improvement in long-term survival after hospital discharge but not in freedom from reoperation after the change from atrial to arterial switch for transposition of the great arteries. AB - OBJECTIVE: To compare survival, freedom from reoperation, and functional status between atrial switch and arterial switch operations for transposition of the great arteries. METHODS: Data from 88, 329, and 512 patients who underwent Mustard, Senning, and arterial switch operations between 1974 and 2006 were analyzed. RESULTS: In-hospital mortalities were 8.0% for Mustard, 4.6% for Senning, and 6.4% for arterial switch. Presence of ventricular septal defect (hazard ratio 3.3, P < .001) was the only risk factor for in-hospital mortality in multivariate analysis. Follow-up for Mustard was 22.6 +/- 8.1 years, for Senning was 18.2 +/- 5.7 years, and for arterial switch was 9.5 +/- 5.7 years. Highest survival at 20 years was after arterial switch (96.6% +/- 1.3%), followed by Senning (92.6% +/- 1.5%) and Mustard (82.4% +/- 4.3%). Transposition with ventricular septal defect (hazard ratio 3.1, P < .001), transposition with ventricular septal defect and left ventricular outflow tract obstruction (hazard ratio 3.0, P = .029), and Mustard operation (hazard ratio 2.1, P = .011) emerged as risk factors for late death, with arterial switch a protective factor (hazard ratio 0.3, P = .010). Highest freedom from reoperation at 20 years was after Senning (88.7% +/- 1.9%), followed by arterial switch (75.0% +/- 6.4%) and Mustard (70.6% +/- 5.4%). Presence of complex transposition (hazard ratio 2.1, P < .001), previous palliative operation (hazard ratio 1.8, P = .016), surgery between 1985 and 1995 (hazard ratio 2.6, P = .002), surgery after 1995 (hazard ratio 3.5, P < .001), and Mustard operation (hazard ratio 3.3, P < .001) emerged as risk factors for reoperation. CONCLUSION: Change from atrial to arterial switch led to improved long-term survival after hospital discharge but not to lower incidence of reoperation. Survival and freedom from reoperation are determined by morphology. PMID- 19185151 TI - Mechanical cavopulmonary assist maintains pulmonary and cerebral blood flow in a piglet model of a bidirectional cavopulmonary shunt with high pulmonary vascular resistance. AB - OBJECTIVES: We tested mechanical cavopulmonary blood flow assist by incorporating a novel miniature centrifugal pump into a 1(1/2)-ventricle type cavopulmonary connection in neonatal pigs. METHODS: Nine 3-week-old piglets (mean body weight, 10.2 kg) were used: mechanical cavopulmonary assist (n = 6) and controls (n = 3). A bidirectional cavopulmonary connection between the superior vena cava and the main pulmonary artery was created. The superior vena cava and pulmonary artery were also connected by cannulas with an interposed centrifugal pump. The cavoarterial mechanical cavopulmonary assist was performed at pump speeds of 1500, 2000, 2500, and 3000 rpm. Retrograde superior vena caval flow was limited by a band on the superior vena cava. A bidirectional cavopulmonary connection was created in the control animals, which then had a pure 1(1/2)-ventricle repair physiology without mechanical support. Hemodynamics, blood gas, and cerebral blood flow measured by ultrasound were analyzed. Catheter-based dilatation of the surgically created superior vena cava obstruction was tested. RESULTS: Incremental increases in pump speed augmented bidirectional cavopulmonary shunt blood flow (P =.03) and diminished superior vena caval pressure (P =.03), thereby improving cerebral perfusion pressure. Pump flow of 3000 rpm was equivalent to baseline superior vena caval flow (before caval flow, 392 +/- 48 mL/min vs MCPA, 371 +/- 120 mL/min; mean +/- SD; P = not significant). The mechanical cavopulmonary assist group had higher Doppler velocities of the middle cerebral artery and higher transcerebral oxygen difference(P < .05) than controls. Balloon dilatation of the superior vena cava band was successful. CONCLUSIONS: Mechanical cavopulmonary assist maintained bidirectional cavopulmonary shunt flow, thereby sustaining primary bilateral cavopulmonary shunt physiology in a neonatal pig model of high pulmonary vascular resistance. The mechanical cavopulmonary assist maintained cerebral blood flow and metabolism with an adequate transcerebral pressure gradient. PMID- 19185153 TI - Mechanical valves versus the Ross procedure for aortic valve replacement in children: propensity-adjusted comparison of long-term outcomes. AB - OBJECTIVE: We aimed to identify characteristics differentiating children undergoing aortic valve replacement by using mechanical prostheses versus the Ross procedure and to compare survival and the need for aortic valve reoperation after each procedure. METHODS: From 1983 to 2004, 346 children underwent aortic valve replacement (215 underwent the Ross procedure and 131 underwent placement of a mechanical prosthesis). Factors associated with procedure choice were used to construct a propensity score for use as a covariate in regression models to adjust for potential confounding by indication. RESULTS: Patients undergoing the Ross procedure were younger, more likely to have a congenital cause, and less likely to have a rheumatic or connective tissue cause. They had a lower frequency of regurgitation, required more annular enlargement, and had less concomitant cardiac surgery. Competing-risk analysis showed that 16 years after aortic valve replacement, 20% of patients had died without subsequent aortic valve replacement, 25% underwent second aortic valve replacement, and 55% remained alive without further replacement. After propensity adjustment, factors associated with early-phase death included mechanical valves and a nonrheumatic cause. Mechanical valves were also associated with constant-phase mortality. Repeated aortic valve replacement was associated with the Ross procedure and a rheumatic cause. Both factors were also associated with all-cause cardiac reoperation. In children receiving mechanical prostheses, younger age and smaller valve size were significant risk factors for death. Freedom from homograft replacement after the Ross procedure was 82% at 16 years of follow-up. CONCLUSION: Results from this study showed good outcomes and an acceptable complication rate with both valve choices. Given the significantly increased risk of early and late death in younger children receiving smaller mechanical valves, the Ross procedure confers survival advantage in this age group at the expense of increased reoperation risk, especially in patients with a rheumatic cause. PMID- 19185155 TI - Biventricular repair in patients with heterotaxy syndrome. AB - OBJECTIVE: Complex intracardiac and extracardiac anatomy is often confronted during biventricular repair in patients with heterotaxy syndrome. We examined factors affecting surgical outcomes in these patients. METHODS: Between January 1990 and July 2007, 371 patients received a diagnosis of heterotaxy syndrome; 91 (91/371, 24.5%) underwent biventricular repair. Left atrial isomerism was present in 73% (66/91) and right atrial isomerism in 10% (9/91), with indeterminate atrial anatomy in 17% (16/91). Median age at biventricular repair was 6.8 months (5 days to 22.3 years). Systemic venous anomalies were present in 75 patients, pulmonary venous anomalies in 26, and endocardial cushion defects in 36. Transposition complexes were present in 15 patients with atrioventricular discordance in 10; 8 underwent double switch, 2 received a physiologic repair, 2 underwent arterial switch, and 3 underwent the Rastelli operation. Other conotruncal anomalies included double-outlet right ventricle in 10 patients, tetralogy of Fallot in 3, and hemitruncus in 2. Separation of systemic from pulmonary venous return included intra-atrial baffling in 48 patients and extracardiac grafting in 2. Combined lesions were common, occurring in 99% (90/91). Statistical analysis with Kaplan-Meier and Cox proportional hazards models were performed. RESULTS: Average follow-up was 44.9 +/- 57.5 months (3 days to 189.3 months). Kaplan-Meier estimated survival was 93.4% at 10 years; unbalanced complete atrioventricular canal was the only risk factor for mortality (P = .006). Subsequent procedures were common with a 10-year freedom from reoperation or reintervention of 38% +/- 7.5%. Arrhythmias occurred in 36 (39.6%) patients; bradyarrhythmia in 27 (29.7%) and tachyarrhythmia in 15 (16.5%). Freedom from any arrhythmia was 53.9% +/- 6.7% at 10 years. CONCLUSIONS: Excellent survival for patients with heterotaxy undergoing biventricular repair can be expected, even for multiple, complex lesions. Reintervention is common, and arrhythmia is a long-term concern. This experience shows that patients with heterotaxy syndrome and complex cardiac anatomy can be considered for biventricular repair. Patients with unbalanced complete atrioventricular canal are a high-risk group for which selection criteria are particularly important. PMID- 19185157 TI - Exercise performance and quality of life following surgical repair of anomalous aortic origin of a coronary artery in the pediatric population. AB - OBJECTIVE: We sought to evaluate exercise performance and quality of life in children after surgical repair of anomalous aortic origin of a coronary artery with an interarterial course. METHODS: Patients who had surgery from October 2001 to January 2007 were eligible for inclusion. Exercise performance and quality of life were prospectively assessed by maximal exercise tests and age-appropriate questionnaires, respectively. We used t tests to compare pre- and postoperative exercise data and quality-of-life scores to published normative data. We performed linear regression analyses to assess associations between demographic, anatomic, and exercise variables and quality-of-life score. RESULTS: Of 25/27 patients, 64% were boys, 68% had anomalous right coronary, 32% were asymptomatic. Average age at surgery was 10.8 (+/-4.1) years; median follow-up was 14.5 (2 to 48) months. Postoperative percent-predicted exercise values were: peak heart rate 97 (+/-6), working capacity 91 (+/-15), maximal oxygen consumption 82 (+/-16). In those who had preoperative exercise testing (n = 11), resting and maximal heart rates decreased significantly without significant change in exercise performance. Average child quality of life was 85/100 (+/-13) and parent-proxy 88 (+/-11) compared with normal scores of 83 (+/-15) and 88 (+/-12), respectively. CONCLUSION: There is mild chronotropic impairment in children and adolescents following anomalous coronary artery repair without a decline in exercise performance. This does not appear to impair their overall quality of life. Because long-term effects on heart rate, exercise performance, and quality of life are unknown, serial exercise tests should be included as routine care of these patients. PMID- 19185158 TI - Parachute mitral valve: morphologic descriptors, associated lesions, and outcomes after biventricular repair. AB - OBJECTIVE: In "true" parachute mitral valve, mitral valve chordae insert into one papillary muscle. In parachute-like asymmetric mitral valve, most or all chordal attachments are to one papillary muscle. This study compared morphologic features, associated lesions, and palliation strategies of the two parachute mitral valve and dominant papillary muscle types and examined interventions and midterm outcomes in patients with biventricular circulation. METHODS: Echocardiography and autopsy databases were reviewed to identify patients with "true" parachute mitral valve or parachute-like asymmetric mitral valve from January 1987 to January 2006. Predictors of palliation strategy in the entire cohort, mitral stenosis on initial echocardiogram, and mortality in the biventricular cohort were determined with logistic regression. RESULTS: Eighty six patients with "true" parachute mitral valve (n = 49) or parachute-like asymmetric mitral valve (n = 37) were identified. Chordal attachments to the posteromedial papillary muscle were more common (73%). The presence "true" parachute mitral valve (P = .008), hypoplastic left ventricle (P < .001), and two or more left-sided obstructive lesions (P = .002) predicted univentricular palliation. Among 49 patients maintaining biventricular circulation at follow-up, 8 died median follow-up 6.4 years (7 days-17.8 years). Multivariate analysis revealed that "true" parachute mitral valve was associated with mitral stenosis on initial echocardiogram (P = .03), and "true" parachute mitral valve (P = .04) and conotruncal anomalies (P = .0003) were associated with mortality. Progressive mitral stenosis was found in 11 patients; 2 underwent mitral valve interventions, and 1 died. CONCLUSION: Nearly two thirds of this parachute mitral valve cohort underwent biventricular palliation. Some progression of mitral stenosis occurred, although mitral valve intervention was rare. "True" parachute mitral valve was associated with mitral stenosis on initial echocardiogram. "True" parachute mitral valve and conotruncal anomalies were associated with mortality in the biventricular population. PMID- 19185159 TI - Evaluation of a novel Y-shaped extracardiac Fontan baffle using computational fluid dynamics. AB - OBJECTIVES: The objective of this work is to evaluate the hemodynamic performance of a new Y-graft modification of the extracardiac conduit Fontan operation. The performance of the Y-graft design is compared to two designs used in current practice: a t-junction connection of the venae cavae and an offset between the inferior and superior venae cavae. METHODS: The proposed design replaces the current tube grafts used to connect the inferior vena cava to the pulmonary arteries with a Y-shaped graft. Y-graft hemodynamics were evaluated at rest and during exercise with a patient-specific model from magnetic resonance imaging data together with computational fluid dynamics. Four clinically motivated performance measures were examined: Fontan pressures, energy efficiency, inferior vena cava flow distribution, and wall shear stress. Two variants of the Y-graft were evaluated: an "off-the-shelf" graft with 9-mm branches and an "area preserving" graft with 12-mm branches. RESULTS: Energy efficiency of the 12-mm Y graft was higher than all other models at rest and during exercise, and the reduction in efficiency from rest to exercise was improved by 38%. Both Y-graft designs reduced superior vena cava pressures during exercise by as much as 5 mm Hg. The Y-graft more equally distributed the inferior vena cava flow to both lungs, whereas the offset design skewed 70% of the flow to the left lung. The 12 mm graft resulted in slightly larger regions of low wall shear stress than other models; however, minimum shear stress values were similar. CONCLUSIONS: The area preserving 12-mm Y-graft is a promising modification of the Fontan procedure that should be clinically evaluated. Further work is needed to correlate our performance metrics with clinical outcomes, including exercise intolerance, incidence of protein-losing enteropathy, and thrombus formation. PMID- 19185160 TI - Anatomic repair for congenitally corrected transposition of the great arteries. AB - OBJECTIVE: Anatomic repair is being actively evaluated as the preferred option for congenitally corrected transposition of the great arteries. We present our 13 year experience with this approach. METHODS: Between May 1994 and September 2007, 68 patients with congenitally corrected transposition of the great arteries underwent anatomic repair. Thirty-one patients (group 1, mean age of 94.8 +/- 42.3 months) underwent a combined Rastelli and atrial switch operation. Thirty seven patients (group 2, mean age of 36.1 +/- 46.9 months) underwent an arterial switch operation and atrial rerouting. Eight patients in group 2 had an intact ventricular septum. RESULTS: Group 1 had 5 early deaths (17%) but no late deaths. Three patients underwent conduit revision at a mean follow-up of 62 months. Group 2 had 5 early deaths (13.5%). There were 4 late reoperations (2 pulmonary baffle revisions, 1 mitral valve replacement, and 1 permanent pacemaker implantation) and 4 late deaths (1 secondary to progressive left ventricular dysfunction, 2 secondary to uncontrolled atrial tachyarrhythmia, and 1 secondary to pulmonary hypertension and right ventricular failure). In group 2, 4 patients have a left ventricular ejection fraction less than 40%, 5 patients have moderate aortic incompetence, 5 patients have symptomatic tricuspid incompetence, 1 patient has tricuspid stenosis, 1 patient has superior cava obstruction, and 3 patients are receiving antiarrhythmic therapy. CONCLUSION: The occurrence of left ventricular dysfunction indicate that anatomic repair in the arterial switch group is still fraught with imperfections. The Rastelli group required conduit revisions but has otherwise performed well. PMID- 19185161 TI - Should endobronchial ultrasonography be part of the thoracic surgeon's armamentarium? AB - OBJECTIVE: The study objective was to determine the clinical usefulness and accuracy of endobronchial ultrasound-guided needle aspiration of mediastinal and hilar lymph nodes. METHODS: A retrospective analysis of a thoracic surgery unit's experience was performed. RESULTS: In a period of 19 months, 75 patients underwent the procedure (mean age = 65.5 +/- 1.6 years; male to female = 2:1) most commonly for mediastinal lymphadenopathy in the setting of diagnosed or suspected lung cancer. It was diagnostic in 68.9% after rapid on-site evaluation and 74.3% after final cytologic examination. The rapid on-site evaluation and final cytology results were discordant in 16.2% (P < .001). In 50 cases, the needle aspirate cytology could be compared with pathology results. The sensitivity and specificity for the diagnosis of cancer were 85% and 100%, respectively. The false-negative rate endobronchial ultrasound cytology was 8.1%. Mediastinal lymph node station 7 was most commonly biopsied. The stations with the highest diagnostic yield were: 11R, 3, 10L, and 7. Of the patients with a positive positron emission tomography scan with suspected clinical stage III lung cancer, cancer was downstaged in 40% after endobronchial ultrasound. CONCLUSION: Endobronchial ultrasound-guided needle aspiration is a clinically useful minimally invasive option for lung cancer staging and evaluation of mediastinal lymphadenopathy. The procedure should be considered complementary to mediastinoscopy. PMID- 19185163 TI - Fatal complications of adult paraesophageal hernia: a population-based study. AB - OBJECTIVES: Data on mortality from paraesophageal hernia are scarce. This study focused on mortality associated with its natural history or conservative treatment. METHODS: For this population-based retrospective study, Finland's administrative databases provided preliminary data. Among 333 patients who died from benign esophageal diseases or hiatal hernias, analysis of medical records led us to include 32. RESULTS: From 1987 through 2001 in Finnish hospitals, 563 patients underwent surgical intervention and 67 underwent conservative treatment for paraesophageal hernia. This hernia caused death (mortality, 0.6/1,000,000 of the adult population; 95% confidence interval, 0-1.8/1,000,000) in 32 patients, 29 (91%) with concomitant diseases. The overall mortality rate for the 563 having surgical treatment was 2.7% (15 patients). Three died after elective repair. Of 67 patients hospitalized for symptomatic paraesophageal hernia and treated conservatively, 11 (16.4%) died in the hospital within a mean of 42 months (range, 2-96 months) from onset of symptoms. Four (13%) deaths might have been prevented by elective surgical intervention. Of the 32 deceased patients, 4 (12.5%) had type II, 16 (50%) had type III, and 9 (28.1%) had type IV hiatal hernias. In 3 (9.4%) patients type remained unknown. Death resulted from incarceration in 24 (75%), complications of surgical intervention in 6 (18.8%), and bleeding ulcer in 2 (6.2%). CONCLUSIONS: Overall, most deaths were related to type III or IV hernias in aged patients with concomitant diseases, with those with severe symptoms requiring hospitalization at significant risk. Except for those at high surgical risk, we recommend repair of the paraesophageal hernia, at least in symptomatic patients. PMID- 19185164 TI - Neutrophil/lymphocyte ratio and its association with survival after complete resection in non-small cell lung cancer. AB - OBJECTIVE: Increasing neutrophil/lymphocyte ratios on preoperative blood tests have been associated with worse survival after resection of colorectal cancer. We sought to determine factors associated with increasing neutrophil/lymphocyte ratios and the stage-adjusted prognostic effect in patients undergoing resection for non-small cell lung cancer. METHODS: We performed a retrospective review of patients undergoing complete resection for non-small cell lung cancer between 1999 and 2005. Data acquisition was through patient medical records, blood results recorded on admission before surgical intervention, and follow-up by National Health Service database searches and hospital records. Cox proportional hazards regression was used to estimate the effect of neutrophil/lymphocyte ratio on stage-adjusted survival. RESULTS: During the study period, 178 patients underwent pulmonary resection. Of 177 patients, the majority were male 104 (59%), with a mean age of 63 years (standard deviation, 10 years). The median follow-up time was 29 months (interquartile range, 8-56 months), and overall survival was 83% and 54% at 1 and 5 years, respectively. Higher stage was the only factor found to be associated with increasing neutrophil/lymphocyte ratios (P = .019). Total white cell count (P = .990) and neutrophil count (P = .490), age (P = .290), and cell type (P = .490) were not significant predictors of mortality. On multivariable analysis after adjusting for stage, increasing neutrophil/lymphocyte ratios (hazard ratio, 1.10; 95% confidence interval, 1.03 1.17; P = .004) remained an independent prognostic indicator. CONCLUSIONS: Increasing preoperative neutrophil/lymphocyte ratios are associated with higher stage but remain an independent predictor of survival after complete resection for primary lung cancer and are a potential biomarker to stratify high risk of death in patients with stage I disease. PMID- 19185165 TI - Prognostic impact of intratumoral vessel invasion in completely resected pathologic stage I non-small cell lung cancer. AB - OBJECTIVE: Intratumoral vessel invasion of non-small cell lung cancer is a readily available tumor-related factor that provides direct evidence of microscopic tumor invasion. We assessed the prognostic influence of intratumoral vessel invasion and its ability to provide a differential prediction of prognosis for completely resected pathologic stage I non-small cell lung cancer. METHODS: We analyzed 258 patients with non-small cell lung cancer who underwent complete resection between January of 1996 and December of 2005 and were diagnosed with pathologic stage I disease. In addition to the conventional staging factors, intratumoral vessel invasion in the primary lesion was histologically evaluated by both hematoxylin-eosin and elastic staining. We examined the significance of intratumoral vessel invasion in prognosis and compared the outcomes between patients with and without this factor with stage IA and IB disease, respectively. RESULTS: Intratumoral vessel invasion was found in 124 patients (48%). Five-year survival of patients with or without intratumoral vessel invasion was 74% and 93%, respectively. On multivariate analysis, intratumoral vessel invasion and pleural invasion were shown to be independent prognostic factors. Subgroup analyses suggested that patients with pathologic-stage IA with intratumoral vessel invasion and patients with pathologic-stage IB with both intratumoral vessel and pleural invasion had significantly worse prognosis than patients with the same pathologic stage without these factors. CONCLUSION: The current study indicated that intratumoral vessel invasion and pleural invasion are independent prognostic factors. Intratumoral vessel invasion status can complement the size dependent TNM staging system in pathologic stage I non-small cell lung cancer. PMID- 19185166 TI - Complete resection of pulmonary inflammatory pseudotumors has excellent long-term prognosis. AB - OBJECTIVE: Pulmonary inflammatory pseudotumor is an uncommon disease, often with a benign presentation. However, invasion of adjacent thoracic organs, local recurrence, and distant metastases have been described, and the best management strategy remains unclear. We present a single large institutional experience in patients with pulmonary inflammatory pseudotumor and propose guidelines for treatment of this patient population. METHODS: A retrospective study was performed to review all patients who underwent resection for pulmonary inflammatory pseudotumor between 1974 and 2007. RESULTS: A total of 25 patients were treated with pulmonary inflammatory pseudotumor at the Marie Lannelongue Hospital. The mean age was 33 years. Two patients were referred after an incomplete resection. One patient presented with cerebral metastasis. We performed a complete resection in all patients: wedge resection (n = 7), lobectomy (n = 6), sleeve arterial lobectomy (n = 1), lobectomy with thoracic inlet exenteration (n = 2), bilobectomy (n = 2), pneumonectomy with brain metastasectomy (n = 1), sleeve pneumonectomy (n = 2), sleeve main bronchus or tracheal resection (n = 2), wedge with sleeve main pulmonary artery resections (n = 1), and sleeve pneumonectomy with esophageal, aortic arch, and right pulmonary artery resection (n = 1). No adjuvant therapy was given to any patients. Postoperative 30-day mortality and morbidity rates were 4% and 8%, respectively. With a mean follow-up of 80 months (range 4-369 months, 100% follow-up), actuarial 10-year survival was 89%. One patient died of an extensive sarcomatous recurrence 2 years after surgery. CONCLUSION: Pulmonary inflammatory pseudotumor is a malignant disease affecting young patients with local invasion, distant metastasis, local recurrence, and sarcomatous degeneration. A complete resection should always be performed at initial presentation because of its high likelihood of cure with aggressive management. PMID- 19185167 TI - Induction chemoradiotherapy facilitates radical resection of T4 non-small cell lung cancer invading the spine. AB - OBJECTIVE: We evaluated the outcome, long-term results, and factors affecting outcome of induction chemoradiotherapy followed by surgical resection for T4 non small cell lung cancer invading the spine. METHODS: Retrospective analysis of 23 consecutive patients undergoing radical vertebral resection for non-small cell lung cancer invading the spine between 1996 and 2007 was performed. In most cases, induction chemoradiotherapy consisted of cisplatin and etoposide followed by 45 Gy of radiation. Surgical resection with vertebrectomy was performed en bloc in either a 1-stage or 2-stage operation. Survival was estimated by Kaplan Meier techniques. The log-rank comparison was used to compare groups. RESULTS: There were 13 men and 10 women with a median age of 61 years (range 32-75). Twenty-two patients had induction chemoradiotherapy and 1 had induction chemotherapy alone. Vertebral resections included 6 total vertebrectomies, 15 hemivertebrectomies, and 2 partial vertebrectomies. Complete resection was achieved in 19 (83%) patients. Two (8.7%) patients died postoperatively. Pathologic complete response was observed in 10 (43%) patients. The 3-year survival was 58% (median follow-up, 34 months). Patients who achieved pathologic complete response or near complete response (viable tumor cells < 1%) demonstrated significantly better survival than those who did not (3-year survival, 92% vs 20%; P = .006). CONCLUSION: Highly selected patients with lung cancer invading the spine can potentially be cured with induction chemoradiation therapy followed by radical en bloc resection of the tumor. A multidisciplinary operative strategy allows a significant chance of achieving complete resection in patients requiring multilevel hemivertebrectomy or total vertebrectomy and an appreciable cure rate. PMID- 19185168 TI - Nonseminomatous germ cell tumors: assessing the need for postchemotherapy contralateral pulmonary resection in patients with ipsilateral complete necrosis. AB - OBJECTIVES: Our objective was to explore the pathologic components of residual masses after primary chemotherapy in patients with metastatic nonseminomatous germ cell tumors. METHODS: A multicenter retrospective study was conducted of 71 patients with thoracic residual masses (39 patients had bilateral lung metastasis) after first-line cisplatin-based chemotherapy for disseminated nonseminomatous germ cell tumors. Among the 71 patients, 52 also had a retroperitoneal lymph node dissection. RESULTS: Pathologic findings in postchemotherapy residual masses included complete necrosis, teratoma, and viable cancer in 31%, 55%, and 14% of patients, respectively. Discordant pathologic findings were evidenced between retroperitoneal lymph node and thoracic (lung or mediastinal lymph nodes) residual masses in 27% of patients. When a bilateral pulmonary resection was performed, only 2 (5%) of 39 patients had discordant histologic findings between the two lungs. Among patients who had necrosis only in residual masses from their first lung (n = 20), 19 (95%) also had necrosis only in contralateral lesions. A single patient had necrosis only in the first lung and some teratoma in the contralateral lung. CONCLUSIONS: This report shows a high rate (95%) of pathologic concordance between the two lungs. Avoiding contralateral lung surgery could therefore be considered when complete necrosis is found in the first lung after induction chemotherapy for nonseminomatous germ cell tumor. PMID- 19185169 TI - A phase I study of extrapleural pneumonectomy and intracavitary intraoperative hyperthermic cisplatin with amifostine cytoprotection for malignant pleural mesothelioma. AB - OBJECTIVE: This study was undertaken to determine maximum tolerated dose and toxicity of intraoperative intracavitary hyperthermic cisplatin perfusion with amifostine after extrapleural pneumonectomy for malignant pleural mesothelioma. METHODS: Patients with mesothelioma were prospectively enrolled. Those with resectable disease received amifostine and 1-hour hyperthermic cisplatin perfusion of ipsilateral hemithorax and abdomen. Morbidity, recurrence, and survival were recorded. RESULTS: Forty-two patients were enrolled; 29 underwent resection (operative mortality 7%, 2/29). Median age was 57 years. Eighteen were in pathologic stage I or II; 11 were in stage III. Median hospitalization was 15 days. Common complications were atrial fibrillation (66%, 19 patients), deep venous thrombosis (31%, 9 patients), and grade 3+ renal toxicity (31%, 9 patients). Feasibility was determined. Renal toxicity was unrelated to cisplatin dose, with no maximum tolerated dose determined. Overall median survival was 17 months (resected 20 months, unresected 10 months). Median survivals were 26 months for patients receiving higher cisplatin doses and 16 months for those receiving lower doses (P = .35). Survival was significantly longer with negative extrapleural nodes (31 vs 14 months, P = .0115) and early stage (all resected 35 months for stage I-II vs 14 months for stage III, P = .0022, epithelial 39 months for stage I-II vs 15 months for stage III, P = .0072). CONCLUSION: Early stage and negative extrapleural lymph nodes were associated with prolonged survival. Single-dose amifostine did not protect adequately against cisplatin-induced renal toxicity. Additional cytoprotective strategies are needed to allow determination of cisplatin maximum tolerated dose. PMID- 19185170 TI - Cerebrovascular response to continuous cold perfusion and hypothermic circulatory arrest. AB - OBJECTIVE: Clinical and laboratory studies have documented changes in cerebrovascular resistance after hypothermic circulatory arrest, both with and without adjunctive cerebral perfusion modalities. This study was designed to clarify whether these changes are due to cerebral edema, resistance vessel abnormalities, or alterations in the cerebral microcirculation. METHODS: Four mature swine underwent hypothermic circulatory arrest for 60 minutes, and 7 mature swine underwent cold cerebral perfusion for 60 minutes to simulate antegrade selective perfusion. All were rewarmed and weaned from cardiopulmonary bypass. Pial vascular diameter and reactivity were measured in vivo through a cranial window and ex vivo in an organ chamber; cerebral microvascular endothelium was studied in culture for release of vasoactive mediators. Cerebral water content was recorded. RESULTS: Cold perfusion caused pial arteriole and venule constriction, whereas hypothermic circulatory arrest alone caused pial arteriole and venule dilatation. Cold perfusion caused a temporal loss of endothelium-dependent vasodilatation, most notably to bradykinin. Hypothermic circulatory arrest caused a loss of nitric oxide-mediated endothelium-dependent vasodilatation. Endothelium-independent vasoreactivity remained intact in both groups. Endothelial cells from the cold group had a vasoconstrictive secretory phenotype, whereas endothelial cells from the hypothermic circulatory arrest group had a vasodilatory phenotype. Cerebral water content was the same in both groups. CONCLUSION: The increase in cerebrovascular resistance observed after cold cerebral perfusion is caused by resistance vessel constriction and may be promoted by an altered microcirculation. Hypothermic circulatory arrest alone is associated with endothelium-dependent vasoparesis. Both could contribute to cerebral injury in the early hours after operation. PMID- 19185171 TI - Chronic partial unloading restores beta-adrenergic responsiveness and reverses receptor downregulation in failing rat hearts. AB - OBJECTIVES: Mechanical unloading with a left ventricular assist device promotes "reverse remodeling," including restoration of beta-adrenergic receptor signaling and function. We compared the effects of partial unloading and complete unloading on beta-adrenergic responsiveness and gene expressions in failing rat hearts by use of heterotopic heart-lung or heart transplantation models. METHODS: Four weeks after ligation of the left anterior descending artery in Lewis rats, rats with heart failure were divided into 3 groups: infarcted hearts and lungs transplanted into the recipient rats (heart failure-partial unloading, n = 8); infarcted hearts transplanted into the recipient rats (heart failure-complete unloading, n = 7); infarcted (heart failure, n = 8) hearts without transplantation. Normal rats (n = 7) were used as controls. Papillary muscle function and gene expressions were studied at 2 or 4 weeks after transplantation. RESULTS: In 2-week models, baseline developed tension of papillary muscles significantly increased in heart failure-partial unloading and heart failure complete unloading compared with heart failure (0.15 +/- 0.07 and 0.12 +/- 0.05 g/mm(2) vs 0.02 +/- 0.01 g/mm(2), P < .05). However, in 4-week models, they decreased to 0.11 +/- 0.03 and 0.10 +/- 0.03 g/mm(2). In 4-week but not in 2-week models, the increase from baseline in baseline developed tension produced by beta adrenergic stimulation (isoproterenol, 10(-8) and 10(-7) mol/L) was significantly increased in heart failure-partial unloading compared with heart failure-complete unloading and heart failure (P < .05). The mRNA expressions of brain natriuretic peptide and beta(1)- and beta(2)-adrenergic receptors were normalized in both 2- and 4-week models of heart failure-partial unloading. CONCLUSIONS: Chronic partial unloading but not complete unloading improved beta-adrenergic responsiveness and normalized brain natriuretic peptide and beta(1)- and beta(2) adrenergic receptor mRNA expressions in the failing rat hearts. PMID- 19185172 TI - Cell-based gene therapy modifies matrix remodeling after a myocardial infarction in tissue inhibitor of matrix metalloproteinase-3-deficient mice. AB - OBJECTIVE: Cell-based gene therapy can enhance the effects of cell transplantation by temporally and spatially regulating the release of the gene product. The purpose of this study was to evaluate transient matrix metalloproteinase inhibition by implanting cells genetically modified to overexpress a natural tissue inhibitor of matrix metalloproteinases (tissue inhibitor of matrix metalloproteinase-3) into the hearts of mutant (tissue inhibitor of matrix metalloproteinase-3-deficient) mice that exhibit an exaggerated response to myocardial infarction. Following a myocardial infarction, tissue inhibitor of matrix metalloproteinase-3-deficient mice undergo accelerated cardiac dilatation and matrix disruption due to uninhibited matrix metalloproteinase activity. This preliminary proof of concept study assessed the potential for cell-based gene therapy to reduce matrix remodeling in the remote myocardium and facilitate functional recovery. METHODS: Anesthetized tissue inhibitor of matrix metalloproteinase-3-deficient mice were subjected to coronary ligation followed by intramyocardial injection of vector-transfected bone marrow stromal cells, bone marrow stromal cells overexpressing tissue inhibitor of matrix metalloproteinase-3, or medium. Functional, morphologic, histologic, and biochemical studies were performed 0, 3, 7, and 28 days later. RESULTS: Bone marrow stromal cells and bone marrow stromal cells overexpressing tissue inhibitor of matrix metalloproteinase-3 significantly decreased scar expansion and ventricular dilatation 28 days after coronary ligation and increased regional capillary density to day 7. Only bone marrow stromal cells overexpressing tissue inhibitor of matrix metalloproteinase-3 reduced early matrix metalloproteinase activities and tumor necrosis factor alpha levels relative to medium injection. Bone marrow stromal cells overexpressing tissue inhibitor of matrix metalloproteinase-3 were also more effective than bone marrow stromal cells in preventing progressive cardiac dysfunction, preserving remote myocardial collagen content and structure, and reducing border zone apoptosis for at least 28 days after implantation. CONCLUSIONS: Tissue inhibitor of matrix metalloproteinase-3 overexpression enhanced the effects of bone marrow stromal cells transplanted early after a myocardial infarction in tissue inhibitor of matrix metalloproteinase-3-deficient mice by contributing regulated matrix metalloproteinase inhibition to preserve matrix collagen and improve functional recovery. PMID- 19185173 TI - Coronary artery bypass grafting with a minimized cardiopulmonary bypass circuit: a prospective, randomized trial. AB - OBJECTIVE: The study was designed to determine differences in blood loss and transfusion associated with a minimized cardiopulmonary bypass circuit versus a standard bypass circuit. METHODS: From February 2005 through April 2006, 199 patients were randomized to undergo coronary artery bypass grafting with a standard cardiopulmonary bypass circuit (Medtronic, Inc., Minneapolis, Minn) or a minimized bypass circuit, the Medtronic Resting Heart Circuit. Laboratory perimeters (hemoglobin and platelet count), were measured at baseline, after initiation of cardiopulmonary bypass, and on intensive care unit admission. Lowest values recorded were noted. Blood administration was controlled by study specific protocol orders, (transfusion for hemoglobin <8mg%). Patient demographic data were retrieved from the Society of Thoracic Surgeons database. Blood product administration was recorded during hospital admission, and chest tube drainage as total output collected from operating room to discontinuation. Continuous variables were tested with a Wilcoxin rank test, and categoric variables with X(2) and Fisher's exact tests. RESULTS: Hematocrit, equivalent at baseline, was higher in minimized circuit cohort at lowest point during cariopulmonary bypass (31.5% +/- 3.9% vs. 25.5% +/- 3.7%), after protamine (31.6% +/- 3.9% vs 29.2% +/- 3.7%), and on intensive care unit arrival (35.2% +/- 4.1% vs 31.8% +/- 3.5%, P < .001). Similarly, platelet count was higher in minimized circuit group on intensive care unit arrival, as was lowest platelet count recorded (170 x 10(3) +/- 48 cells/mm(3) vs 107 x 10(3) +/- 28 cells/mm(3), P < .0001). Time to extubation was shorter in minimized circuit group (848 +/- 737 minutes vs. 526 +/ 282 minutes, (P < .01), and total chest tube drainage was lower (1124 +/- 647 mL vs. 506 +/- 214 mL, P < .01). Fewer red blood cells (148 vs 19 units) were given in minimized circuit group (P < .0001). CONCLUSIONS: A minimized cardiopulmonary bypass circuit provides less hemodilution, platelet consumption, chest tube output and lower post-operative blood loss than standard cardiopulmonary bypass. Red blood cell usage was also less. All differences are advantageous. PMID- 19185174 TI - A mouse model of orthotopic, single-lung transplantation. AB - OBJECTIVES: Progress in studying acute and chronic pulmonary allograft rejection has been hampered by the lack of feasible experimental animal transplantation models. Contemporary approaches are limited by anatomic applicability (heterotopic tracheal implantation) and lack of genetic variability (rat model). To utilize the breadth of available genetic modifications in a physiologic setup, we optimized and validated a procedure of orthotopically transplanted, perfused, and ventilated single pulmonary transplantation in mice. METHODS: C57BL/6 mice served as recipient, with Balb/c as donor. At time of harvest, explanted lungs were perfused with Perfadex, and the heart-lung block excised. Under 30 to 40x magnification, vessels and bronchus were cuffed. Following left thoracotomy in the recipient, hilar structures were incised and cuff-anastomosed with the corresponding donor parts. Allogeneic and syngeneic transplantations (n = 12/group) were performed with a follow-up period of 5 days and up to 90 days, respectively. RESULTS: The success rate of lung transplantation in mice was 87.5% (21/24). Mean cold ischemia time was 32.3 +/- 3.7 minutes, and warm ischemia time was 30.8 +/- 9.5 minutes. Deaths were due to bleeding during dissection of the hilus and/or caused by thrombosis postoperatively. Allogeneic grafts were rejected by day 5; syngeneic grafts were slightly congested but mainly unchanged up to day 90 posttransplantation. CONCLUSIONS: Unilateral lung transplantation in mice can be performed in a standardized and controlled fashion with low mortality, comparable to the rat. Employing transgenic and knockout mice strains, this procedure holds great promise to advance the understanding of immunologic pathways in acute and chronic rejection in a physiologic model of pulmonary transplantation. PMID- 19185175 TI - Cellular electrophysiologic and mechanical evidence of superior vascular protection in pulmonary microcirculation by Perfadex compared with Celsior. AB - OBJECTIVE: Pulmonary endothelial function is critical in posttransplant lung performance. Hyperkalemic organ preservation solutions alter vascular endothelial function through the non-nitric oxide and non-prostacyclin pathway, but the most frequently used lung preservation solutions, Perfadex (Vitrolife Sweden, Kungsbacka, Sweden) (K(+) 6 mmol/L) and Celsior (IMTIX SangStat Company, Lyon, France) (K(+) 15 mmol/L), have not been evaluated on pulmonary endothelial protection. We compared the non-nitric oxide and non-prostacyclin-mediated endothelial function in porcine pulmonary microarteries of lung preserved by Perfadex or Celsior solution at 4 degrees C for 4 hours. METHODS: The non-nitric oxide and non-prostacyclin-mediated endothelial function was determined by measuring the membrane potential in a single pulmonary smooth muscle cell (group II, n = 6) and bradykinin-induced relaxation (group I, n = 8) in pulmonary microarteries preserved in Krebs (a, control), Perfadex (b), or Celsior (c), with inhibitors of nitric oxide and prostacyclin. RESULTS: Membrane potential hyperpolarization decreased in IIc (4.5 +/- 0.2 mV, P < .05) but was preserved (P > .05) in IIa (6.6 +/- 0.1 mV) and IIb (6.3 +/- 0.3 mV). Resting membrane potential was depolarized in IIc (-42.8 +/- 1.3 mV) compared with IIa (-58.7 +/- 0.6 mV) and IIb (-56.7 +/- 0.9 mV) (P < .05). Hyperpolarization-associated relaxation (37.3% +/- 7.2% vs 59.7% +/- 7.7%) and sensitivity (EC(50)) (-7.29 +/- 0.13 vs -7.75 +/- 0.06 log M) to bradykinin significantly (P < .05) decreased in Ic but not in Ia and Ib. CONCLUSION: This in vitro study simulating clinical conditions demonstrates that Perfadex preserves endothelium-dependent smooth muscle relaxation and hyperpolarization better than Celsior solution in regard to the electrophysiologic and mechanical properties observed at cellular and vascular levels. This study provides a new method at the level of basic science to evaluate the solutions for heart/lung preservation. PMID- 19185176 TI - Giant primary paraganglioma of the left ventricle. PMID- 19185177 TI - A case of antiphospholipid syndrome presenting with a floating thrombus in the ascending aorta. PMID- 19185178 TI - Midterm patency rate after saphenous vein grafting with a PAS-Port device. PMID- 19185179 TI - Late rupture of polytetrafluoroethylene neochordae after mitral valve repair. PMID- 19185180 TI - Stapler versus precision dissection and sealant in comparing completion technique of fissures for lobectomy: True or statistical advantages? PMID- 19185183 TI - Periodic paralysis. AB - Periodic paralyses are rare diseases characterized by severe episodes of muscle weakness concomitant to variations in blood potassium levels. It is thus usual to differentiate hypokalemic, normokalemic, and hyperkalemic periodic paralysis. Except for thyrotoxic hypokalemic periodic paralysis and periodic paralyses secondary to permanent changes of blood potassium levels, all of these diseases are of genetic origin, transmitted with an autosomal-dominant mode of inheritance. Periodic paralyses are channelopathies, that is, diseases caused by mutations in genes encoding ion channels. The culprit genes encode for potassium, calcium, and sodium channels. Mutations of the potassium and calcium channel genes cause periodic paralysis of the same type (Andersen-Tawil syndrome or hypokalemic periodic paralysis). In contrast, distinct mutations in the muscle sodium channel gene are responsible for all different types of periodic paralyses (hyper-, normo-, and hypokalemic). The physiological consequences of the mutations have been studied by patch-clamp techniques and electromyography (EMG). Globally speaking, ion channel mutations modify the cycle of muscle membrane excitability which results in a loss of function (paralysis). Clinical physiological studies using EMG have shown a good correlation between symptoms and EMG parameters, enabling the description of patterns that greatly enhance molecular diagnosis accuracy. The understanding of the genetics and pathophysiology of periodic paralysis has contributed to refine and rationalize therapeutic intervention and will be without doubts the basis of further advances. PMID- 19185184 TI - Myotonia congenita. AB - Myotonia is a symptom of many different acquired and genetic muscular conditions that impair the relaxation phase of muscular contraction. Myotonia congenita is a specific inherited disorder of muscle membrane hyperexcitability caused by reduced sarcolemmal chloride conductance due to mutations in CLCN1, the gene coding for the main skeletal muscle chloride channel ClC-1. The disorder may be transmitted as either an autosomal-dominant or recessive trait with close to 130 currently known mutations. Although this is a rare disorder, elucidation of the pathophysiology underlying myotonia congenita established the importance of sarcolemmal chloride conductance in the control of muscle excitability and demonstrated the first example of human disease associated with the ClC family of chloride transporting proteins. PMID- 19185185 TI - Familial hemiplegic migraine. AB - Migraine is a severely debilitating episodic disorder affecting up to 12% of the general population. Migraine arises from both genetic and environmental factors, complicating our understanding of what makes the migraine brain susceptible to attacks. In recent years, powerful genetic screening tools have revealed several single genes linked to migraine. One example of a monogenic subtype of migraine is familial hemiplegic migraine (FHM), a rare form of migraine with aura. The fact that FHM and common multifactorial migraine have many overlapping clinical features indicates that they likely share underlying pathophysiological pathways. In addition, the identification of monogenic subtypes has made it possible to generate suitable animal models for migraine. The purpose of this review is to present an overview of the clinical features of migraine and discuss the continuing highway of migraine gene discovery. The genes associated with FHM will be discussed, including what we have learned from studying the functional consequences of FHM mutations in cellular and animal models. PMID- 19185186 TI - Genetics and molecular pathophysiology of Na(v)1.7-related pain syndromes. AB - SCN9A, the gene which encodes voltage-gated sodium channel Na(v)1.7, is located on human chromosome 2 within a cluster of other members of this gene family. Na(v)1.7 is present at high levels in most peripheral nociceptive neurons in dorsal root ganglion (DRG) and in sympathetic neurons. In addition to its focal tissue-specific expression, Na(v)1.7 is distinguished by its ability to amplify small depolarizations, thus acting as a threshold channel and modulating excitability. Dominantly inherited gain-of-function mutations in SCN9A have been linked to two familial painful disorders: inherited erythromelalgia (IEM) and paroxysmal extreme pain disorder (PEPD). One set of mutations leads to severe episodes of pain in the feet and hands in patients with IEM, and a different set of mutations causes pain in a perirectal, periocular, and mandibular distribution in patients with PEPD. These mutations allow mutant channels to activate in response to weaker stimuli, or to remain open longer in response to stimulation. The introduction of mutant channels into DRG neurons alters electrogenesis and renders these primary sensory neurons hyperexcitable. Mutant Na(v)1.7 channels lower the threshold for single action potentials and increase the number of action potentials that neurons fire in response to suprathreshold stimuli. In contrast, recessively inherited loss-of-function mutations in SCN9A, which cause a loss of function of Na(v)1.7 in patients, lead to indifference to pain with sparing of motor and cognitive abilities. The central role of Na(v)1.7 in these disorders, and the apparently limited consequences of loss of this channel in humans make it an attractive target for treatment of pain. PMID- 19185187 TI - Channelopathies of transepithelial transport and vesicular function. AB - The transport of ions across cellular membranes is crucial for various functions, including the transport of salt and water across epithelia, the control of electrical excitability of muscle and nerve, and the regulation of cell volume or the acidification and ionic homeostasis of intracellular organelles. This review will focus on ion channel diseases (channelopathies) that are due to impaired transepithelial transport or due to mutations in vesicular ion transporters or channels. When needed for a more comprehensive understanding of organ function, also other ion transport diseases will be mentioned. PMID- 19185188 TI - Preface. Veterinary public health. PMID- 19185189 TI - Public health for the twenty-first century: what role do veterinarians in clinical practice play? AB - All veterinarians, regardless of their formal job description, serve the public good and contribute to public health. The public health activities veterinarians engage in most frequently in clinical practice are in the areas of disease detection, reporting, and prevention. This article provides a brief overview of the basic functions of public health, while emphasizing the public health roles that veterinary clinicians play in their day-to-day practice of veterinary medicine and how they might extend their interest and involvement in this field. The multidisciplinary nature of the field of public health and the benefits of collaboration with other health care and public health professionals are also discussed. PMID- 19185190 TI - Disease reporting and surveillance: where do companion animal diseases fit in? AB - Disease surveillance and reporting is a necessary and integral part of public health practice. Surveillance systems have been developed over many years for both human medicine and veterinary medicine. However, these systems are not usually interconnected. Today, with the benefits of advanced information technology, the development and integration of existing and new resources in companion-animal practice should be focused on "one medicine-one health" for the betterment and health of all species. This means more sharing of surveillance data, greater cooperation among organizations involved in surveillance, and further integration of human and animal surveillance activities. PMID- 19185191 TI - Companion animals as sentinels for public health. AB - Animal sentinel surveillance is a key component of public health risk assessment. While many species serve as animal sentinels, companion animals have an especially valuable role as sentinels because of their unique place in people's lives, with exposure to similar household and recreational risk factors as those for the people who own them. Dogs and cats can help in early identification of food contamination, infectious disease transmission, environmental contamination, and even bioterrorism or chemical terrorism events. Early detection, leading to early intervention, can minimize the impact of these adverse events on both animal and human health. PMID- 19185192 TI - Influenza in dogs and cats. AB - Influenza has been long absent from the list of infectious diseases considered as possibilities in dogs and cats. With the discovery that avian influenza H5N1 can infect cats and dogs, and the appearance of canine influenza H3N8, small animal veterinarians have an important role to play in detection of influenza virus strains that may become zoonotic. Small animal veterinarians must educate staff and clients about influenza to improve understanding as to when and where influenza infection is possible, and to avert unreasonable fears. PMID- 19185193 TI - Emerging tick-borne diseases. AB - Ticks are capable of transmitting numerous pathogens to both humans and their pets. The risks of tick-borne disease risks vary geographically and are determined by the climate, environment, the presence of rodents and other mammal reservoirs, and the species of ticks parasitizing wild and domestic animals. Zoonoses such as Lyme borreliosis, tularemia, and tick-borne rickettsioses can emerge in previously nonendemic areas when circumstances favorable to their maintenance and transmission arise. Tick-borne zoonosis can be prevented by implementation and adoption of an integrated program to reduce the likelihood of tick bites on pets and their owners. PMID- 19185194 TI - Pets and antimicrobial resistance. AB - Antimicrobial resistance is a growing problem and is a significant public health issue. An increasing number of organisms are developing resistance to many of the antimicrobial agents available for treatment of infections in both humans and animals. These resistant organisms often result in greater disease severity, longer hospitalization, and increased care and treatment costs. This article reviews the current situation of antimicrobial resistance in companion small animals and highlights how important it is for veterinarians to recognize the significance of antimicrobial resistance and to commit to the judicious use of antimicrobial agents. PMID- 19185195 TI - The human-companion animal bond: how humans benefit. AB - The human-animal bond is extremely important to most clients of small animal veterinary practices. Pet ownership, or just being in the presence of a companion animal, is associated with health benefits, including improvements in mental, social, and physiologic health status. This article provides the research data regarding the human health benefits of companion animals, animal-assisted therapy, animal-assisted activities, and assistance animals; reviews measures that can be taken to enable safe pet ownership for the immunocompromised, and discusses the veterinarian's role in supporting immune-compromised clients and clients who have assistance animals. Client education and enhanced veterinary care can reduce the risk from zoonotic diseases, even for the immunocompromised. PMID- 19185196 TI - The impact of companion animal problems on society and the role of veterinarians. AB - The benefits of companion animals are immense, but there can be negative impacts also. Noise, destructive behaviors, excrement, bites, and the overpopulation of domestic cats and dogs are some of the major problems that can result in stress and hardships on owners, neighbors, the community, and the pets themselves. The perpetuation of pets in society requires that the negative aspects of living with dogs and cats be addressed. Veterinarians can play an important role in addressing these problems by incorporating the concept of behavior wellness into their practices and promoting education regarding husbandry, animal behavior, responsible pet ownership, and the effects of pets on the environment. PMID- 19185197 TI - Emergency management during disasters for small animal practitioners. AB - This article provides a broad overview of emergency management during disasters, including its organizational structure and the emergency management cycle. It delineates activities that small animal clinicians might engage in with regards to disaster mitigation, preparedness, response, and recovery. It also introduces such concepts as the incident command system and the national incident management system. Last, this article provides some suggestions for how small animal veterinarians might seek further training and education in this increasingly important field. PMID- 19185198 TI - Border health: who's guarding the gate? AB - Changes in the global trade market have led to a thriving international pet trade in exotic animals, birds, and puppies. The flood of animals crossing the United States' borders satisfies the public demand for these pets but is not without risk. Imported pets may be infected with diseases that put animals or the public at risk. Numerous agencies work together to reduce the risk of animal disease introduction, but regulations may need to be modified to ensure compliance. With more than 280,000 dogs and 183,000 wildlife shipments being imported into the United States each year, veterinarians must remain vigilant so they can recognize potential threats quickly. PMID- 19185199 TI - Local veterinary diagnostic laboratory, a model for the one health initiative. AB - The San Diego County Animal Disease Diagnostic Laboratory (ADDL) is unique in its emphasis on protecting both human and animal health in San Diego County, and its use of interagency and community collaboration to create strong, effective public health programs. This article describes the ADDL core programs of avian and vector-borne disease surveillance, rabies testing, and animal abuse investigations and uses selected case studies to illustrate the need for a local veterinary diagnostic laboratory to safeguard the health of humans and animals. The ADDL serves as a role model for other local communities to develop vital public health partnerships to ensure a healthier community. PMID- 19185200 TI - [The second life of daptomycin: analysis of a successful come-back]. PMID- 19185201 TI - [Enterococcus faecalis endocarditis successfully treated with daptomycin]. PMID- 19185202 TI - [The role of antibiotic-therapy referral in therapeutic innovation. Apropos of a case of streptococcal endocarditis]. PMID- 19185203 TI - [Conservative treatment of a hip prosthesis infection due to methicillin resistant Staphylococcus aureus using prolonged antibiotic therapy based on linezolid followed by daptomycin]. PMID- 19185204 TI - [Treatment of mitral MRSA endocarditis using daptomycin]. PMID- 19185205 TI - Conditioned preferences and aversions in infant rats mediated through ethanol inhalation. AB - Little is known about the acute motivational effects of inhaled ethanol during early postnatal development. We analyzed the motivational properties of ethanol inhalation in infant rats by using two distinct schedules of ethanol vapor delivery. Ethanol was presented in a continuous conditioning trial or in separate, distributed trials. Maximum blood ethanol concentrations (BECs) induced by these schedules were 55 and 15 mg%, respectively (Experiment 1). In Experiment 2, subjects were given daily pairings (postnatal days [PD] 14 and 15) between a tactile conditioned stimulus (CS, sandpaper) and the postabsorptive effects of ethanol inhalation. A tactile preference test (PD16) revealed a significant aversion for the CS in pups given continuous exposure to ethanol vapor. In Experiment 3, an ethanol pre-exposure phase (PD13) preceded tactile-ethanol pairings. During conditioning, pups were given distributed pairings between the tactile CS and ethanol or uncontaminated air. At test, ethanol-pre-exposed animals spent significantly more time on the ethanol-related CS than on an alternative texture. These results indicate that inhaled ethanol exerts differential hedonic effects in infant rats as a function of schedules of exposure that yield different levels of intoxication. Continuous experience with ethanol vapor induces aversive learning. Yet, pre-exposure to ethanol vapor allowed expression of ethanol-induced appetitive learning in pups given distributed vapor ethanol exposure. PMID- 19185206 TI - Ethanol induces locomotor activating effects in preweanling Sprague-Dawley rats. AB - Abuse of drugs exerts biphasic motor activity effects, which seem to be associated with their motivational effects. In the case of ethanol, heterogenous rat strains appear to be particularly sensitive to the sedative effects of the drug. In contrast, ethanol's activating effects have been consistently reported in rats genetically selected for ethanol affinity. Heightened ethanol affinity and sensitivity to ethanol's reinforcement are also observed in nonselected rats during early ontogeny. In the present study, we examined psychomotor effects of ethanol (1.25 and 2.5 g/kg) in 8-, 12-, and 15-day-old pups. Motor activity in a novel environment was assessed 5-10 or 15-20 min following drug treatment. Rectal temperatures and latency to exhibit the righting reflex were recorded immediately after locomotor activity assessment. Ethanol exerted clear activating effects at 8 and 12 days of age (Experiments 1a and 1b) and to a lesser extent at 15 days. At this age, ethanol enhanced locomotor activity in the first testing interval (Experiment 1b) and suppressed locomotion at 15-20 min (Experiment 1a). Ethanol mediated motor impairment was more pronounced in the youngest group (postnatal day 8) than in the older ones. Blood ethanol concentrations were equivalent in all age groups. The present study indicates that preweanling rats are sensitive to ethanol's stimulating effects during the second postnatal week, and suggest that specific periods during early ontogeny of the rat can provide a valuable framework for the study of mechanisms underlying ethanol's stimulation and reinforcement effects. PMID- 19185207 TI - Chronic ethanol and withdrawal effects on kainate receptor-mediated excitatory neurotransmission in the rat basolateral amygdala. AB - Withdrawal (WD) anxiety is a significant factor contributing to continued alcohol abuse in alcoholics. This anxiety is extensive, long-lasting, and develops well after the obvious physical symptoms of acute WD. The neurobiological mechanisms underlying this prolonged WD-induced anxiety are not well understood. The basolateral amygdala (BLA) is a major emotional center in the brain and regulates the expression of anxiety. New evidence suggests that increased glutamatergic function in the BLA may contribute to WD-related anxiety following chronic ethanol exposure. Recent evidence also suggests that kainate-type ionotropic glutamate receptors are inhibited by intoxicating concentrations of acute ethanol. This acute sensitivity suggests potential (KA-R) contributions by these receptors to the increased glutamatergic function seen during chronic exposure. Therefore, we examined the effect of chronic intermittent ethanol (CIE) and WD on KA-R-mediated synaptic transmission in the BLA of Sprague-Dawley rats. Our study showed that CIE, but not WD, increased synaptic responses mediated by KA-Rs. Interestingly, both CIE and WD occluded KA-R-mediated synaptic plasticity. Finally, we found that BLA field excitatory postsynaptic potential responses were increased during CIE and WD via a mechanism that is independent of glutamate release from presynaptic terminals. Taken together, these data suggest that KA-Rs might contribute to postsynaptic increases in glutamatergic synaptic transmission during CIE and that the mechanisms responsible for the expression of KA-R dependent synaptic plasticity might be engaged by chronic ethanol exposure and WD. PMID- 19185208 TI - Excessive alcohol consumption is blocked by glial cell line-derived neurotrophic factor. AB - We previously found that activation of the glial cell line-derived neurotrophic factor (GDNF) pathway in the ventral tegmental area (VTA) reduces moderate alcohol (ethanol) intake in a rat operant self-administration paradigm. Here, we set out to assess the effect of GDNF in the VTA on excessive voluntary consumption of ethanol. Long-Evans rats were trained to drink large quantities of a 20% ethanol solution in an intermittent-access two-bottle choice drinking paradigm. The rats were given three 24-h sessions per week, and GDNF's actions were measured when rats achieved a baseline of ethanol consumption of 5.5g/kg/24h. We found that microinjection of GDNF into the VTA 10min before the beginning of an ethanol-drinking session significantly reduced ethanol intake and preference, but did not affect total fluid intake. We further show that GDNF greatly decreased both the first bout of excessive ethanol intake at the beginning of the session, and the later consummatory activity occurring during the dark cycle. These data suggest that GDNF is a rapid and long-lasting inhibitor of "binge-like" ethanol consumption. PMID- 19185210 TI - Chronic ethanol consumption resulting in the downregulation of insulin receptor beta subunit, insulin receptor substrate-1, and glucose transporter 4 expression in rat cardiac muscles. AB - The objective of this study was to investigate the effect of chronic ethanol intake on the expression of insulin receptor beta subunit (IR-beta), insulin receptor substrate-1 (IRS-1), and glucose transporter 4 (Glut4) in rat cardiac muscle. Forty-eight male Wistar rats were randomly classified into four groups and to each group, ethanol was administered daily at the respective doses of 0 (control, C), 0.5 g kg(-1) (low ethanol, L), 2.5 g kg(-1) (middle ethanol, M), and 5 g kg(-1) (high ethanol, H). Twenty-two weeks later, the rats were anesthetized, and the left ventricle muscles were excised. The IR-beta, IRS-1, and Glut4 mRNA levels in the cardiac muscle tissues were detected by reverse transcription polymerase chain reaction (RT-PCR); the IR-beta, tyrosine phosphorylation of IR-beta (PY-IR-beta), IRS-1, tyrosine phosphorylation of IRS-1 (PY-IRS-1), and Glut4 protein levels were measured by Western blotting. Compared to the control group, the IR-beta, IRS-1, and Glut4 mRNA levels in groups H and M decreased remarkably. In addition, the protein levels of IR-beta, IRS-1, and Glut4 showed a corresponding decline in ethanol-treated groups, especially in group H. Moreover, the PY-IR-beta and PY-IRS-1 protein levels decreased in the hearts of ethanol-treated rats with corresponding changes in the IR-beta and IRS 1 protein levels. The present study showed that chronic ethanol intake could downregulate the expression levels of IR-beta, IRS-1, and Glut4 in rat cardiac muscles. PMID- 19185211 TI - Effect of exercise training on ethanol-induced oxidative damage in aged rats. AB - It is well known that lipid peroxidation increases with age, and alcohol drinking further exacerbates this damage. The present study determined the effect of regular exercise training on alcohol-induced oxidative damage and antioxidant status in the liver of aged animals. The age-matched Wistar albino rats (3 months young, n=24; 18 months old, n=24) were evenly divided into four groups: control (C), exercise trained (Ex), ethanol drinking (Et), and exercise plus ethanol drinking (Ex+Et). With ethanol drinking, hepatic malondialdehyde (MDA) level was significantly elevated above control (P<.001), whereas glutathione (GSH) and ascorbic acid (vitamin C) contents were significantly decreased below control. These changes were found to be greater in the aged rats than those of the young rats. For both age groups, exercise training significantly reversed the increase in MDA and decreases in GSH and ascorbic acid induced by ethanol drinking. The present study showed that ethanol-induced deterioration in lipid peroxidation and reduction in antioxidant status in the liver were exacerbated with age. Here, we found that exercise training significantly reversed the adverse conditions that were caused by ethanol in aged rats. PMID- 19185209 TI - Ethanol disrupts NMDA receptor and astroglial EAAT2 modulation of Kv2.1 potassium channels in hippocampus. AB - Delayed-rectifier Kv2.1 channels are the principal component of voltage-sensitive K+ currents (I(K)) in hippocampal neurons and are critical regulators of somatodendritic excitability. In a recent study, we demonstrated that surface trafficking and phosphorylation of Kv2.1 channels is modulated by NMDA-type glutamate receptors and that astroglial excitatory amino acid transporters 2 (EAAT2) regulate the coupling of NMDA receptors and Kv2.1 channels. Because ethanol is known to acutely inhibit NMDA receptors, we sought to determine if NMDA receptor and astroglial EAAT2 modulation of Kv2.1 channels is impaired by ethanol in the rodent hippocampus. As expected, bath application of NMDA to hippocampal cultures reduced the size of Kv2.1 clusters and produced a hyperpolarizing shift in the voltage-dependent activation of I(K) that was associated with dephosphorylated Kv2.1 channels. Ethanol, applied acutely, prevented the hyperpolarizing shift in activation of I(K) induced by NMDA and restored Kv2.1 clustering and phosphorylation to near control levels. Ethanol also attenuated the dephosphorylation of Kv2.1 channels produced by the EAAT2 selective inhibitor dihydrokainic acid. These data demonstrate that acute ethanol disrupts changes in Kv2.1 channels that follow NMDA receptor activation and impairs astroglial regulation of the functional coupling between NMDA receptors and Kv2.1 channels. PMID- 19185212 TI - Alcohol consumption and lipodystrophy in HIV-infected adults with alcohol problems. AB - Lipodystrophy is a common long-term complication of HIV infection that may lead to decreased quality of life and less adherence to antiretroviral therapy (ART). A complete understanding of the etiology of HIV-associated lipodystrophy has not yet been achieved, although factors related to the virus, per se, and use of ART appear to be related. Alcohol use is common among HIV-infected patients and has biological effects on fat distribution, yet alcohol's relationship to HIV associated lipodystrophy has not been examined. The goal of this clinical study was to assess the effect of alcohol consumption on lipodystrophy in HIV-infected adults with alcohol problems. This was a prospective study (2001-2006) of 289 HIV infected persons with alcohol problems. The primary outcome was self-reported lipodystrophy, which was assessed at one time point (median 29 months after enrollment). Alcohol use was assessed every 6 months and classified as: abstinent at all interviews; > or = 1 report of moderate drinking but no heavy drinking; 1 or 2 reports of heavy drinking; or > or = 3 reports of heavy drinking. Multivariable logistic regression models were fit to the data. Fifty-two percent (150/289) of subjects reported lipodystrophy. Alcohol consumption was: 34% abstinent at all interviews; 12% > or = 1 report of moderate drinking, but no heavy drinking; 34% 1-2 reports of heavy drinking; and 20% > or = 3 reports of heavy drinking. Although not statistically significant, subjects with alcohol use had a higher odds of lipodystrophy (adjusted odds ratios and 95% confidence interval: > or = 1 report of moderate drinking, 2.36 [0.89, 6.24]; 1-2 reports of heavy drinking, 1.34 [0.69, 2.60]; > or = 3 reports of heavy drinking, 2.07 [0.90, 4.73]). Alcohol use may increase the odds of developing HIV-associated lipodystrophy among subjects with alcohol problems. However, larger studies are needed to fully elucidate the role and impact of alcohol consumption on the development of this common long-term complication of HIV infection and its treatment. PMID- 19185213 TI - HTR3B is associated with alcoholism with antisocial behavior and alpha EEG power- an intermediate phenotype for alcoholism and co-morbid behaviors. AB - Alcohol use disorders (AUD) with co-morbid antisocial personality disorder (ASPD) have been associated with serotonin (5-HT) dysfunction. 5-HT3 receptors are potentiated by ethanol and appear to modulate reward. 5-HT3 receptor antagonists may be useful in the treatment of early-onset alcoholics with co-morbid ASPD. Low voltage alpha electroencephalogram (EEG) power, a highly heritable trait, has been associated with both AUD and ASPD. A recent whole genome linkage scan in one of our samples, Plains American Indians (PI), has shown a suggestive linkage peak for alpha power at the 5-HT3R locus. We tested whether genetic variation within the HTR3A and HTR3B genes influences vulnerability to AUD with comorbid ASPD (AUD+ASPD) and moderates alpha power. Our study included three samples: 284 criminal alcoholic Finnish Caucasians and 234 controls; two independent community ascertained samples with resting EEG recordings: a predominantly Caucasian sample of 191 individuals (Bethesda) and 306 PI. In the Finns, an intronic HTR3B SNP rs3782025 was associated with AUD+ASPD (P=.004). In the Bethesda sample, the same allele predicted lower alpha power (P=7.37e(-5)). Associations between alpha power and two other HTR3B SNPs were also observed among PI (P=.03). One haplotype in the haplotype block at the 3' region of the gene that included rs3782025 was associated with AUD+ASPD in the Finns (P=.02) and with reduced alpha power in the Bethesda population (P=.00009). Another haplotype in this block was associated with alpha power among PI (P=.03). No associations were found for HTR3A. Genetic variation within HTR3B may influence vulnerability to develop AUD with comorbid ASPD. 5-HT3R might contribute to the imbalance between excitation and inhibition that characterize the brain of alcoholics. PMID- 19185214 TI - Balancing act. PMID- 19185215 TI - Fluoroquinolones and postoperative endophthalmitis. PMID- 19185216 TI - Fluoroquinolones and postoperative endophthalmitis. PMID- 19185218 TI - Retinal phototoxicity in the aging pseudophakic and phakic eye. PMID- 19185220 TI - Formula for determining corneal refractive power. PMID- 19185222 TI - Endothelial cell damage with phacoemulsification techniques. PMID- 19185223 TI - Consultation section. Cataract surgical problem. PMID- 19185233 TI - Simple method to evaluate adequacy of capsule for foldable intraocular lens implantation in the sulcus. AB - We describe a simple method for evaluating the adequacy of the capsule for implantation of a foldable intraocular lens (IOL) during phacoemulsification complicated by a posterior capsule tear. After cortical lens material clean-up and anterior vitrectomy (if required), 3 knots of 10-0 nylon are tied to the trailing haptic of the IOL using a 3/1/1 configuration. Then, the IOL is inserted into the sulcus. If it is unstable when the surgeon tries to deepen or shallow the anterior chamber, gently pulling the nylon thread will prevent posterior IOL dislocation. Two further attempts to implant the IOL in the other meridians of the capsule are made. The results of using this technique in 10 eyes of 10 patients are reported. Optic decentration occurred in 1 case (10%). No postoperative dislocation was observed, and no patient required surgery for IOL decentration during the follow-up. PMID- 19185234 TI - Alternative technique for implantation of a scleral-fixated intraocular lens. AB - We describe an alternative method of implanting a scleral-fixated intraocular lens (IOL) that facilitates passage of the suture and improves control of suture placement. An ab externo approach introduces a loop of polypropylene (Prolene) through the sclera in a single 27-gauge puncture. The loop is used to secure the IOL haptic in a cow-hitch fashion, which minimizes the risk for the knot to unravel and the IOL to dislocate. After the IOL is placed in the eye, it is secured with the externalized curved suture needle. Knots are covered with a scleral flap. This technique improves efficiency and control in placing an IOL near the normal anatomical location of the crystalline lens when the absence of capsule support precludes nontethered placement of an IOL in the sulcus or capsular bag. PMID- 19185235 TI - Triamcinolone-assisted vitrectomy in pediatric cataract surgery. AB - We describe a technique to manage anterior vitreous face disturbance that may occur during performance of a manual posterior continuous curvilinear capsulorhexis (PCCC) in pediatric eyes. The technique renders the vitreous visible to the anterior segment surgeon and ensures a thorough and complete anterior vitrectomy in congenital cataract surgery after a manual PCCC using preservative-free triamcinolone acetonide. PMID- 19185236 TI - Postoperative corneal shape changes: microincision versus small-incision coaxial cataract surgery. AB - PURPOSE: To compare changes in corneal topography and in regular and irregular astigmatism after coaxial clear corneal microincision cataract surgery (MICS) and after coaxial small-incision cataract surgery (SICS). SETTING: Hayashi Eye Hospital, Fukuoka, Japan. METHODS: Induced corneal astigmatism was determined using vector analysis. The averaged corneal shape changes and degree of irregular astigmatism were examined using videokeratography preoperatively as well as 2 days and 1, 2, 4, and 8 weeks postoperatively. RESULTS: One hundred twenty eyes of 60 patients scheduled for phacoemulsification were evaluated. Sixty eyes had MICS (2.00 mm), and the 60 contralateral eyes had SICS (2.65 mm). The mean induced corneal astigmatism was significantly less in the MICS group than in the SICS group (P .05). The mean IOP remained unchanged during the entire follow-up (P> .05). The most frequent complication was postoperative pupillary block in 7 eyes (17.5%). Two eyes with severe glare and 1 eye with unexpected myopia and discomfort had pIOL explantation. Exchange of the pIOL was performed in 2 eyes and laser-assisted subepithelial keratectomy in 1 eye to treat unexpected postoperative myopia. No pIOL-induced glaucoma or cataract developed. CONCLUSIONS: Refractive stability and predictability were high at the 1-year follow-up. There was no gain in corrected visual acuity. Despite 2 iridotomies performed 2 weeks preoperatively, the main complication was early pupillary block. PMID- 19185239 TI - Objective and subjective preoperative refraction techniques for wavefront optimized and wavefront-guided laser in situ keratomileusis. AB - PURPOSE: To determine which of 3 methods for measuring preoperative refractive error yields the best refractive outcomes after wavefront-guided (Visx Star S4) or wavefront-optimized (WaveLight Allegretto Wave) excimer laser in situ keratomileusis (LASIK). SETTING: Emory Eye Center and Emory Vision, Emory University, Atlanta, Georgia, USA. METHODS: This retrospective analysis of LASIK, performed from June to December 2007, analyzed sphere, cylinder, and spherical equivalent (SE) refractions generated from 3 methods (manifest refraction, aberrometer autorefraction [CustomVue WaveScan], corneal analyzer autorefraction [Nidek ARK-10000 OPD]), actual programmed treatment, and absolute deviation from ideal treatment (deviation) for each technique. RESULTS: In the wavefront-guided group (63 eyes, 33 patients), manifest refraction and aberrometer autorefraction generated similar deviation for sphere and SE; both were significantly better than corneal analyzer autorefraction (P= .02 and P= .03, respectively). Aberrometer autorefraction generated less cylinder deviation than the other methods (both P= .003). In the wavefront-optimized group (61 eyes, 36 patients), manifest refraction generated less deviation for sphere and SE than aberrometer autorefraction or corneal analyzer autorefraction (sphere: P= .005 and P= .009, respectively; SE: P= .005 and P= .002, respectively). Manifest refraction and aberrometer autorefraction cylinder generated similar deviation, while aberrometer autorefraction was less than corneal analyzer autorefraction (P= .041). CONCLUSIONS: Overall, manifest refraction was most accurate in generating postoperative emmetropia with both laser treatments; however, aberrometer autorefraction produced the least cylinder deviation for wavefront-guided treatment. Corneal analyzer autorefraction produced the least accurate results with both lasers. PMID- 19185240 TI - Higher-order aberrations after wavefront-optimized photorefractive keratectomy and laser in situ keratomileusis. AB - PURPOSE: To analyze the changes in higher-order aberrations (HOAs) that occur after wavefront-optimized photorefractive keratectomy (PRK) and laser in situ keratomileusis (LASIK). SETTING: Emory Eye Center and Emory Vision, Atlanta, Georgia, USA. METHODS: This retrospective analysis comprised eyes that had PRK or LASIK from June 2004 through October 2005. Postoperative outcome measures included 3-month uncorrected visual acuity (UCVA), best spectacle-corrected visual acuity (BSCVA), manifest refraction spherical equivalent (MRSE), changes in the root mean square (RMS) and grouped coefficient HOAs (microns) measured with a corneal analyzer, and subjective assessment of visual aberrations. RESULTS: One hundred consecutive eyes of 54 patients had PRK, and 100 contemporaneous consecutive eyes of 71 patients had LASIK. The PRK and LASIK populations were similar in general demographics, preoperative HOAs, and postoperative UCVA and BSCVA. The mean MRSE was slightly hyperopic after PRK (mean +0.11 diopters [D]) and slightly myopic after LASIK (mean -0.19 D) (P< .0001). There were no statistically significant changes in RMS or grouped coefficient HOA values after PRK or LASIK, nor were there significant differences in postoperative RMS or grouped coefficient HOA values between PRK and LASIK. One percent of PRK and LASIK patients reported a subjective increase in postoperative visual aberrations; 5% reported a subjective improvement postoperatively. CONCLUSIONS: Wavefront-optimized excimer laser surgery did not induce significant HOAs after PRK or LASIK. The 2 techniques were equally efficacious and had equivalent postoperative HOA profiles. PMID- 19185241 TI - Randomized intraindividual comparison of posterior capsule opacification between a microincision intraocular lens and a conventional intraocular lens. AB - PURPOSE: To evaluate the differences in posterior capsule opacification (PCO) and visual and optical performance between a microincision intraocular lens (IOL) and a conventional IOL. SETTING: Ophthalmology Department, St. Thomas' Hospital, London, United Kingdom. METHODS: Patients with bilateral cataract were prospectively randomized to receive a HumanOptics MC611MI microincision IOL (microlens group) or an Alcon AcrySof MA60AC 3-piece IOL (control group) in either eye and were followed for 24 months. Best corrected visual acuity (BCVA) (logMAR) was measured; PCO was quantified by POCO software analysis of retroillumination images. Aberrations and modulation transfer function (MTF) were measured at the 24-month visit. RESULTS: The study enrolled 32 patients. The mean percentage area PCO was greater in the microlens group than in the control group from 3 months onward and was statistically significant from 12 months onward. The greatest difference in PCO between groups was at 24 months: mean 25.45%+/-34.51% (SD) in the microlens group versus 7.82%+/-13.35% in the control group (P= .029). The BCVA in the control group was slightly better at all time points; the difference between groups was statistically significant at 3, 6, and 12 months. No significant difference in aberrations was detected. The MTF curves were comparable for both IOLs. CONCLUSIONS: Both IOLs provided good visual performance. There was no evidence of distortion of the microincision IOL in the capsular bag. The microincision IOL had poorer PCO performance, which was visually significant and was caused by migration of lens epithelial cells through its broad optic-haptic junctions. PMID- 19185242 TI - Optimizing the intracameral dilation regimen for cataract surgery: prospective randomized comparison of 2 solutions. AB - PURPOSE: To compare the efficacy of epi-Shugarcaine solution (epinephrine 0.025% and lidocaine 0.75% in fortified balanced salt solution [BSS Plus]) (epi-S) and Lundberg and Behndig's intracameral dilation solution (cyclopentolate 0.1%, phenylephrine 1.5%, and lidocaine 1%) (LB) for intracameral dilation for cataract surgery. SETTING: Private clinic, Lincolnwood, Illinois, USA. METHODS: In this paired-eye single-masked prospective study, the first eye scheduled for surgery was randomized to receive LB solution or epi-S solution intracamerally at the time of cataract surgery; the second eye to have surgery received the alternative solution. Each eye received 1 drop of tropicamide 1% at least 20 minutes before the start of surgery. Pupil measurements were made before the incision was created, 1 minute after intracameral solution injection, after the anterior chamber was filled with an ophthalmic viscosurgical device, and at the conclusion of the case. Pupil size was objectively graded via software operated by a video technician who was masked to which solution was used. RESULTS: Eighty-four eyes of 42 patients were enrolled in the study. At each time point after instillation, the pupil was statistically significantly larger with epi-S solution than with LB solution, averaging 0.528 mm larger 1 minute after instillation (mean 7.183 versus 6.665 mm) to 0.34 mm larger (mean 7.381 versus 7.040 mm) by the end of the case. CONCLUSION: When combined with 1 drop of tropicamide 1% preoperatively, both formulations provided safe and effective intracameral dilation for cataract surgery; however, the epi-S solution was the more efficacious at each step of the procedure. PMID- 19185243 TI - Mydriasis with different preparations of topically administered lidocaine hydrochloride. AB - PURPOSE: To quantify the mydriatic effect and side effects of topical lidocaine hydrochloride with different pH values and concentrations in healthy volunteers. SETTING: Umea University Hospital Eye Clinic, Umea, Sweden. METHODS: In this intraindividual comparison double-masked randomized study, healthy volunteers were given topical lidocaine 8%, lidocaine 8% pH 6.0, or a placebo 3 times at 90 second intervals. Thereafter, a drop of tropicamide 0.5% or placebo was given (tropicamide setting). Another group of healthy volunteers was given topical lidocaine 4%, 8%, or 16% 3 or 6 times at 90-second intervals (dose-response setting). Before and 15, 30, 60, and 180 minutes after eyedrop instillation, the near point was determined and Scheimpflug photography was performed. RESULTS: The tropicamide setting comprised 26 volunteers (mean age 23.5 years) and the dose response setting, 10 volunteers (mean age 24.5 years). The direct mydriatic effect of lidocaine was larger at pH 6.0 (mean peak effect 0.61+/-0.06 [SEM] versus 0.42+/-0.04 mm) (P< .05), and the augmentation of tropicamide mydriasis was larger at pH 6.0. Lidocaine 8% had a better direct mydriatic effect than lidocaine 4% and lidocaine 16% (mean peak effect 0.60+/-0.09 mm versus 0.19+/ 0.08 mm and 0.28+/-0.10 mm, respectively) (P< .01). Lidocaine 16% given 6 times caused corneal side effects. CONCLUSIONS: Lidocaine 8% with pH 6.0 had a maximum direct mydriatic effect and gave maximum augmentation of tropicamide mydriasis without causing unwanted corneal side effects and thus should be preferable for topical anesthesia in cataract surgery. PMID- 19185244 TI - Objective accommodation measurements in pseudophakic subjects using an autorefractor and an aberrometer. AB - PURPOSE: To compare objective measures of accommodation with the WR-5100K autorefractor and iTrace aberrometer to subjective push-down and defocus tests in normal phakic subjects and pseudophakic subjects with standard monofocal intraocular lenses (IOLs). SETTING: University of Houston, College of Optometry, Houston, Texas, USA. METHODS: The push-down test and defocus curves were used to stimulate and measure accommodation subjectively in pseudophakic subjects. For objective testing, a near target was pushed up and refraction measured objectively. For comparison, the same objective measurements were performed in phakic subjects. Calibration tests were performed with soft contact lenses in phakic subjects with varying refractive error and in pseudophakic subjects. RESULTS: Fifteen phakic subjects (mean age 28.9 years+/-5.52 [SD]) and 10 pseudophakic subjects (mean age 66.2+/-11.23 years) participated. Subjectively measured accommodative amplitude in the pseudophakic group was 3.28+/-1.11 diopters (D) in the right eye and 3.64+/-1.38 D in the left eye. Defocus curves in the pseudophakic group had a range of 2.00 D for distance-corrected visual acuity of 20/40 or better. Objective measurements in the pseudophakic group were 0.11+/-0.50 D with the autorefractor and 0.10+/-0.47 D with the aberrometer. Contact lens calibrations showed good agreement with the 1:1 line. CONCLUSIONS: The subjective tests overestimated accommodative amplitude relative to the objective measures. The autorefractor and aberrometer were accurate, reliable, and appropriate for objective accommodation measurements in pseudophakes. Objective accommodation measurements such as these can be used to evaluate the performance of accommodating IOLs. PMID- 19185245 TI - Ultrasound biomicroscopic evaluation of the efficacy of a transillumination technique for ciliary sulcus localization in transscleral fixation of posterior chamber intraocular lenses. AB - PURPOSE: To evaluate the efficacy of the transillumination technique for precisely locating the ciliary sulcus in transscleral fixation of posterior chamber intraocular lenses (PC IOLs) by determining the haptic positions with ultrasound biomicroscopy (UBM). SETTING: Department of Ophthalmology, Numune Training and Research Hospital, Ankara, Turkey. METHODS: Ultrasound biomicroscopy was used to determine the haptic positions in eyes with ab externo transsclerally fixated PC IOLs. Eyes were randomly assigned to a control group, in which transscleral fixation of a PC IOL was performed, or an endoilluminator-assisted group, in which transscleral fixation was combined with transillumination. RESULTS: The study evaluated 33 eyes of 28 patients ranging in age from 16 to 81 years. The control group comprised 19 eyes (17 patients) and the endoilluminator assisted group, 14 eyes (12 patients). All haptics were easily visualized with UBM. The UBM examination showed that the rate of haptics located in the sulcus was statistically significantly higher in the endoilluminator-assisted group (64%) than in the control group (24%) (P= .001). There was no significant difference in either group in the rate of precise sulcus location between the straight needle and the 28-gauge insulin needle (P> .05). CONCLUSIONS: Ultrasound biomicroscopy showed the difficulty in reliably suturing the haptics in the ciliary sulcus, even with the use of a transillumination technique. However, the results suggest that the transillumination technique is a safe and easy procedure and helps the surgeon identify the ciliary sulcus during transscleral fixation of PC IOLs more precisely than without the use of transillumination. PMID- 19185246 TI - Optical quality in eyes with aspheric intraocular lenses and in younger and older adult phakic eyes: comparative study. AB - PURPOSE: To compare ocular higher-order aberrations (HOAs) in eyes with aspheric intraocular lenses (IOL) and in phakic eyes. SETTING: Department of Ophthalmology, Military Health Service Institute, Warsaw, Poland. METHODS: Higher order aberrations with a 5.0 mm pupil and best spectacle-corrected visual acuity (BSCVA) were analyzed in a young phakic group (age 20 to 40 years), older phakic group (age 40 to 60 years), and a pseudophakic group (age 62 to 84 years; SN60WF aspheric IOL). RESULTS: The mean coma was 0.13 microm+/-0.07 (SD) in the younger phakic group, 0.18+/-0.08 microm in the older phakic group, and 0.16+/-0.08 microm in the pseudophakic group; the difference between the 2 phakic groups was statistically significant (P< .005). The mean spherical aberration was 0.07+/ 0.05 microm, 0.11+/-0.05 microm, and 0.06+/-0.04 microm, respectively; the only statistically significant difference was between the 2 phakic groups (P< .005). The mean HOA value was 0.54+/-0.18 microm, 0.76+/-0.12 microm, and 0.92+/-0.31 microm, respectively; the difference was statistically significant between the 2 phakic groups or between the younger phakic group and the pseudophakic group (both P< .005). There were no statistically significant differences in BSCVA between the 3 groups. CONCLUSIONS: Spherical and coma aberrations were similar in eyes with an aspheric IOL and younger phakic eyes, although HOAs were higher in pseudophakic eyes. There were no differences between pseudophakic eyes and older phakic eyes in coma and HOA, although pseudophakic eyes had lower spherical aberration. PMID- 19185247 TI - Intermediate optimization of vision with bilateral nonaspheric multifocal intraocular lens implantation. AB - PURPOSE: To evaluate binocular visual function, refraction, and subjective outcomes of intermediate optimization of vision after simultaneous bilateral implantation of the nonaspheric AcrySof SA60D3 ReSTOR multifocal intraocular lens (IOL). SETTING: Private clinic, Abbotsford, British Columbia, Canada. METHODS: Intermediate optimization was performed by overcorrection of 1.00 diopter (D) to induce mild myopia in the nondominant eye in patients having bilateral implantation of the multifocal IOL. Distance manifest refraction and visual acuity at several distances were determined; contrast sensitivity function (CSF), near-point stereo acuity, and subjective outcomes were assessed postoperatively at 2 weeks and 1, 3, 6, and 12 months. RESULTS: Thirty-two patients with intended intermediate optimization and 5 patients with unintended intermediate optimization were prospectively evaluated. The mean manifest refraction spherical equivalent was stable from 2 weeks (-0.94 D+/-0.22 [SD]) to 12 months (-0.93+/ 0.23 D) in the intermediate optimization eyes. Overall, postoperative mean binocular uncorrected near, intermediate, and distance visual acuities were 20/23, 20/23, and 20/22, respectively. No significant differences were found between best distance-corrected and uncorrected stereo acuity or between best distance-corrected and uncorrected binocular CSF under most conditions (P> .05). On questioning, 97% of patients had little or no difficulty seeing and were bothered occasionally or never by the visual fluctuation between the near range and intermediate range. CONCLUSION: Intermediate optimization with slight myopia in 1 eye after bilateral implantation of the multifocal IOL offered consistent good vision at all ranges, indicating it a safe option for patients who require good vision at intermediate ranges. PMID- 19185249 TI - Effect of trypan blue staining on the elastic modulus of anterior lens capsules of diabetic and nondiabetic patients. AB - PURPOSE: To determine whether trypan blue causes a significant difference in the biomechanical properties (stiffness) of diabetic and nondiabetic anterior lens capsules and to determine whether diabetes significantly alters the stiffness of anterior lens capsules. SETTING: Veterans Administration Boston Healthcare Center System, Jamaica Plain, Massachusetts, USA. METHODS: In this unmasked prospective randomized controlled experimental study, anterior lens capsules were obtained from diabetic and nondiabetic patients approved for cataract surgery. Four treatment groups were created: (1) capsules of nondiabetic patients without trypan blue; capsules of nondiabetic patients with trypan blue; (3) capsules of diabetic patients without trypan blue; (4) capsules of diabetic patients with trypan blue. Anterior lens capsule stiffness as a function of elastic (Young's) modulus (kilopascals) was measured with the Hysitron TriboIndenter. RESULTS: The use of trypan blue led to significantly stiffer anterior lens capsules (P= .036). Trypan blue had the greatest effect on the stiffness of diabetic lens capsules (P= .046). CONCLUSION: Trypan blue contributed to increased capsule stiffness during capsulorhexis, especially in diabetic patients. PMID- 19185248 TI - Use of cobalt blue light in grading cataract and estimating vision loss. AB - PURPOSE: To test an anecdotally reported cataract grading system predictive of a 20/50 visual threshold in cataract-induced vision loss using cobalt blue light. SETTING: Hennepin County Medical Center, Minneapolis, Minneapolis VA Medical Center, Minneapolis, and Regions Hospital, St. Paul, Minnesota, USA. METHODS: Four observers evaluated pure nuclear cataracts using a standardized cobalt-blue light protocol. Observers graded a nuclear cataract as positive if the posterior capsule was visualized with cobalt blue light and negative if the posterior capsule was not visualized. Results of the grading were compared with cataract induced vision loss in an attempt to establish a threshold for lost visual acuity in grading cataracts with cobalt blue light. The study design was prospective observation of a cohort with a visually significant cataract. RESULTS: This study did not show a clear visual acuity threshold for cataract-induced vision loss using a standardized cobalt-blue-light protocol. Overall, 26.3% (95% confidence interval, 13.4-40.2) of all 20/40 or less dense nuclear cataracts had visible posterior capsules using cobalt blue light, with good estimated interobserver agreement. CONCLUSIONS: Although cobalt blue light is selectively absorbed by yellow pigment in an aging nuclear cataract, its ability to predict visual acuity loss due to lens opacity was limited. PMID- 19185250 TI - Comparison of partial coherence interferometry and ultrasound for anterior segment biometry. AB - PURPOSE: To assess the performance of a partial coherence interferometry (PCI) based device for the determination of anterior segment biometry. SETTING: Clinica Centrofama, Cartagena, Murcia, Spain. METHODS: Central corneal thickness (CCT), anterior chamber depth (ACD), and lens thickness (LT) were measured with the ACMaster PCI anterior segment biometer and an Echoscan US-1800 ultrasound (US) biometer/pachymeter with and without cycloplegia. To determine the precision of the instruments, the same examiner took 30 consecutive CCT, ACD, and LT measurements in a single subject under the same conditions and with and without cycloplegia. The same measurements were performed in additional subjects. RESULTS: Twenty-one eyes (16 subjects) were evaluated. Repeated measurements of the single subject yielded a standard deviation of 4.0 microm for CCT, 106.0 microm for ACD, and 323.0 microm for LT; there were many peaks, mainly in the last 10 readings. There was a high correlation between CCT measurements with both systems with and without cycloplegia (r(2)>0.93), with the US system giving higher values. Differences were significant (P< .001), but not consistent, throughout the range of corneal thicknesses and were greater for thicker corneas. Differences in ACD and LT measurements were similar. Agreement between systems in ACD and LT measurements was considerably lower than for CCT measurements. CONCLUSIONS: The PCI biometer provided precise CCT measurements. The ACD and LT measurements had a higher variance. Differences in CCT measurements between the 2 systems were greater for thicker corneas, with higher values with the US system. PMID- 19185251 TI - Scheimpflug photography keratometry readings for routine intraocular lens power calculation. AB - PURPOSE: To prospectively evaluate keratometry (K) values obtained by Scheimpflug photography in eyes scheduled for cataract surgery, compare the results with K values obtained with an autokeratometer (automated K), and evaluate the K values in commonly used intraocular lens (IOL) power calculation formulas for routine cataract surgery. SETTING: Private clinical ophthalmology practice, Lynwood, California, USA. METHODS: The mean simulated K power (simulated K), equivalent K (equivalent K), and true net power (true net K) readings from the Pentacam Comprehensive Eye Scanner were compared with the automated K readings. Automated K, simulated K, and equivalent K values were compared in commonly used IOL power calculation formulas. RESULTS: The mean automated K value was 43.49 diopters (D)+/-1.75 (SD) and the mean simulated K value, 43.49+/-2.00 D (P> .1). The mean equivalent K value was 43.78+/-1.97 D and exceeded the mean automated K and simulated K by 0.29 D (P> .1). The mean true net K was 42.31+/-2.13 D, which was 1.18 D lower than the automated K and simulated K values (P= .015). The IOL prediction mean absolute error was 0.41+/-0.27 D using the automated K method, 0.50+/-0.36 D using the simulated K method (difference 0.09 D) (P> .1), and 0.65+/-0.35 D using the equivalent K method (difference 0.24 D) (P< .01). CONCLUSION: The K values from Scheimpflug photography did not improve accuracy over autokeratometer values for routine IOL power calculation. PMID- 19185252 TI - Phacoemulsification in eyes with extreme axial myopia. AB - PURPOSE: To determine the outcomes of phacoemulsification of age-related cataract in eyes with extreme axial myopia. SETTING: Moorfields Eye Hospital NHS Foundation Trust, London, United Kingdom. METHODS: This consecutive case series of eyes with an axial length (AL) greater than 30.0 mm was identified retrospectively. Surgical procedure details, complications, and refractive outcome were recorded. The incidence of postoperative retinal detachment (RD) was assessed in a subgroup of eyes without previous RD or vitrectomy. RESULTS: One hundred seventy-seven eyes (124 patients) were identified from a dataset of 6882 cataract procedures; 21 eyes had previous RD or vitrectomy. Intraoperative complications were posterior capsule tear (5 eyes, 2.8%), zonular dehiscence (3 eyes, 1.7%), and anterior capsule tear (2 eyes, 1.1%). At a median follow-up of 25.4 months, 2 (1.3%) of 156 eyes without previous RD or vitrectomy developed RD. The median biometry prediction error in the 156 eyes (88.1%) with an intraocular lens (IOL) was 0.88 diopter (D) (range 0.00 to 5.75 D); 83 eyes (53.2%) were within +/-1.00 D. The median biometry prediction error was greater in the 61 eyes that received a negative-power IOL (1.38 D) than in the 95 eyes that received a plano or positive-power IOL (0.68 D) (P<.001). CONCLUSIONS: The incidence of complications after phacoemulsification in eyes with extreme axial myopia was similar to that in previously reported unselected case series. The refractive outcome showed a hyperopic error that increased with AL, an effect that was primarily the result of the eyes that received a negative-power IOL. PMID- 19185253 TI - Role of depressive symptoms in patient satisfaction with visual quality after laser in situ keratomileusis. AB - PURPOSE: To assess the role of depressive symptoms in patient satisfaction with visual quality after laser in situ keratomileusis (LASIK). SETTING: Navy Refractive Surgery Center, Ophthalmology Department, Naval Medical Center, San Diego, California, USA. METHODS: This was a prospective study of active-duty military patients having bilateral conventional LASIK to treat low to moderate myopia with or without astigmatism. Patients completed the Armstrong Laboratory Personality Survey Depression scale preoperatively. Patients were also given a 10 point Navy questionnaire that included an assessment of patient satisfaction with visual quality at the 1- and 6-month follow-up examinations. Patients were stratified by their visual quality rating after LASIK as highly satisfied, satisfied, and less satisfied and by their depression scale score into low, medium, and high groups. A proportional odds model was developed to compare the visual quality rating between the 3 depression groups 1 and 6 months post LASIK. RESULTS: The study comprised 370 patients. An uncorrected visual acuity of at least 20/20 was achieved in 88.6% of eyes. Patients with higher levels of depressive symptoms had 3.0 times higher odds (95% confidence interval [CI], 1.2 to 7.4) of being less satisfied than those with low levels of depressive symptoms 1 month after LASIK (P 0.69). Other interventions were not found to reduce either incidence or severity of BP episodes compared with controls. Negative trials included five trials of education, four of lumbar supports, two of shoe inserts, and four of reduced lifting programs. CONCLUSIONS: Twenty high-quality controlled trials found strong, consistent evidence to guide prevention of BP episodes in working-age adults. Trials found exercise interventions effective and other interventions not effective, including stress management, shoe inserts, back supports, ergonomic/back education, and reduced lifting programs. The varied successful exercise approaches suggest possible benefits beyond their intended physiologic goals. LEVEL OF EVIDENCE: Systematic review Level I evidence. PMID- 19185281 TI - Mutations in SPINT2 cause a syndromic form of congenital sodium diarrhea. AB - Autosomal-recessive congenital sodium diarrhea (CSD) is characterized by perinatal onset of a persistent watery diarrhea with nonproportionally high fecal sodium excretion. Defective jejunal brush-border Na(+)/H(+) exchange has been reported in three sporadic patients, but the molecular basis of the disease has not been elucidated. We reviewed data from a large cohort of CSD patients (n = 24) and distinguished CSD associated with choanal or anal atresia, hypertelorism, and corneal erosions--i.e., a syndromic form of CSD--occurring in ten families from an isolated form--i.e., classic CSD--presenting in seven families. Patients from both groups have a high risk of mortality due to immediate electrolyte imbalances and complications from long-term parenteral nutrition in the first years of life, but survivors can eventually adapt to partial or complete enteral nutrition. A genome-wide SNP scan was applied and identified a homozygous c.593 1G-->A splicing mutation in SPINT2, encoding a Kunitz-type serine-protease inhibitor, in one extended kindred with syndromic CSD. The same mutation and four distinct, homozygous or compound heterozygous mutations (p.Y163C, c.1A-->T, c.337+2T-->C, c.553+2T-->A) were identified in all syndromic patients. No SPINT2 mutations were found in classic-CSD patients. SPINT2 mutations were associated with loss of protein synthesis or failure to inhibit the serine protease trypsin in vitro. We delineate syndromic CSD as a distinct disease entity caused by SPINT2 loss-of-function mutations. SPINT2 mutations might lead to an excess of yet unknown serine protease activity in affected tissues. PMID- 19185282 TI - A multiplex human syndrome implicates a key role for intestinal cell kinase in development of central nervous, skeletal, and endocrine systems. AB - Six infants in an Old Order Amish pedigree were observed to be affected with endocrine-cerebro-osteodysplasia (ECO). ECO is a previously unidentified neonatal lethal recessive disorder with multiple anomalies involving the endocrine, cerebral, and skeletal systems. Autozygosity mapping and sequencing identified a previously unknown missense mutation, R272Q, in ICK, encoding intestinal cell kinase (ICK). Our results established that R272 is conserved across species and among ethnicities, and three-dimensional analysis of the protein structure suggests protein instability due to the R272Q mutation. We also demonstrate that the R272Q mutant fails to localize at the nucleus and has diminished kinase activity. These findings suggest that ICK plays a key role in the development of multiple organ systems. PMID- 19185283 TI - Searching for genotype-phenotype structure: using hierarchical log-linear models in Crohn disease. AB - There has been considerable recent success in the detection of gene-disease associations. We consider here the development of tools that facilitate the more detailed characterization of the effect of a genetic variant on disease. We replace the simplistic classification of individuals according to a single binary disease indicator with classification according to a number of subphenotypes. This more accurately reflects the underlying biological complexity of the disease process, but it poses additional analytical difficulties. Notably, the subphenotypes that make up a particular disease are typically highly associated, and it becomes difficult to distinguish which genes might be causing which subphenotypes. Such problems arise in many complex diseases. Here, we concentrate on an application to Crohn disease (CD). We consider this problem as one of model selection based upon log-linear models, fitted in a Bayesian framework via reversible-jump Metropolis-Hastings approach. We evaluate the performance of our suggested approach with a simple simulation study and then apply the method to a real data example in CD, revealing a sparse disease structure. Most notably, the associated NOD2.908G-->R mutation appears to be directly related to more severe disease behaviors, whereas the other two associated NOD2 variants, 1007L-->FS and 702R-->W, are more generally related to disease in the small bowel (ileum and jejenum). The ATG16L1.300T-->A variant appears to be directly associated with only disease of the small bowel. PMID- 19185285 TI - Perioperative beta-blockers for major noncardiac surgery: Primum Non Nocere. AB - Recent studies have called into question the benefit of perioperative beta blockade, especially in patients at low to moderate risk of cardiac events. Once considered standard of care, the role of beta-blocker therapy now lies mired in conflicting data that are difficult to apply to the at-risk patient. We provide an overview of the evolution of perioperative beta blockade, beginning with the physiology of the adrenergic system, with emphasis on the biologic rationale for the perioperative implementation of beta-blockers. Although initial studies were small in size and statistically limited, early data showed cardiac benefit with the use of perioperative beta-blockers. However, larger, more recent studies now suggest a lack of benefit and potential harm from this practice. This paradigm holds true especially in those at low-to-moderate cardiovascular risk profiles. Potential explanations for these paradoxical results are discussed, stressing the key differences between earlier and current studies that may explain these divergent outcomes. We conclude by commenting on performance measures as they relate to perioperative beta-blockers and make recommendations for the continued safe implementation of this practice. PMID- 19185284 TI - Common variation in the beta-carotene 15,15'-monooxygenase 1 gene affects circulating levels of carotenoids: a genome-wide association study. AB - Low plasma levels of carotenoids and tocopherols are associated with increased risk of chronic disease and disability. Because dietary intake of these lipid soluble antioxidant vitamins is only poorly correlated with plasma levels, we hypothesized that circulating carotenoids (vitamin A-related compounds) and tocopherols (vitamin E-related compounds) are affected by common genetic variation. By conducting a genome-wide association study in a sample of Italians (n = 1190), we identified novel common variants associated with circulating carotenoid levels and known lipid variants associated with alpha-tocopherol levels. Effects were replicated in the Women's Health and Aging Study (n = 615) and in the alpha-Tocopherol, beta-Carotene Cancer Prevention (ATBC) study (n = 2136). In meta-analyses including all three studies, the G allele at rs6564851, near the beta-carotene 15,15'-monooxygenase 1 (BCMO1) gene, was associated with higher beta-carotene (p = 1.6 x 10(-24)) and alpha-carotene (p = 0.0001) levels and lower lycopene (0.003), zeaxanthin (p = 1.3 x 10(-5)), and lutein (p = 7.3 x 10(-15)) levels, with effect sizes ranging from 0.10-0.28 SDs per allele. Interestingly, this genetic variant had no significant effect on plasma retinol (p > 0.05). The SNP rs12272004, in linkage disequilibrium with the S19W variant in the APOA5 gene, was associated with alpha-tocopherol (meta-analysis p = 7.8 x 10(-10)) levels, and this association was substantially weaker when we adjusted for triglyceride levels (p = 0.002). Our findings might shed light on the controversial relationship between lipid-soluble anti-oxidant nutrients and human health. PMID- 19185286 TI - Erythropoietin induction by electroconvulsive seizure, gene regulation, and antidepressant-like behavioral effects. AB - BACKGROUND: The neuroprotective and trophic actions of erythropoietin (EPO) have been tested in several animal models of insult, injury, and neurodegeneration. Recent studies in human volunteers demonstrated that EPO improves cognition and also elicits antidepressant effects. It is believed that the behavioral effects are mediated by EPO's trophic effect on neuronal systems. We therefore tested whether EPO is able to alter behavior and brain gene expression in rats. METHODS: The expression of EPO and EPO receptor (EPOR) in multiple brain regions was examined by quantitative polymerase chain reaction, in situ hybridization, and immunohistochemistry. The regulation of EPO and the transcription factor hypoxia induced factor-alpha (HIF1alpha) after electroconvulsive seizure (ECS) was investigated. Behavioral effects of EPO were tested in the rodent forced swimming and novelty-induced hypophagia (NIH) models. EPO gene profiles were obtained by microarray analysis of the hippocampus after intracerebroventricular infusion. RESULTS: EPO and EPOR were widely expressed in the brain albeit at low levels. Highest level of EPO and EPOR were in the choroid plexus and striatum, respectively. Peripheral administration of EPO was sufficient to produce a robust antidepressant-like effect in the forced swim and NIH tests. Gene expression profiles revealed that EPO induces the expression of neurotrophic genes such as brain-derived neurotrophic factor, VGF (nonacronymic), and neuritin. CONCLUSIONS: EPO is induced by ECS and independently exhibits antidepressant-like efficacy in the forced swim and NIH tests. EPO regulates the expression of genes implicated in antidepressant action and appears to be a candidate molecule for further testing in neuropsychiatry. PMID- 19185288 TI - Synthesis of the beta-1,3-glucan, laminarahexaose: NMR and conformational studies. AB - The synthesis of laminarahexaose is described. NMR studies of several of the intermediates leading to the beta-1,3-glucan show anomalously small coupling constants for some of the C-1 hydrogens. An X-ray structure for the protected hexasaccharide shows that the small coupling constants are due to some of the glucopyranose rings adopting a twist-boat conformation. The X-ray studies also explain other unexpected NMR observations. PMID- 19185287 TI - Tumor necrosis factor antagonism normalizes rapid eye movement sleep in alcohol dependence. AB - BACKGROUND: In alcohol dependence, markers of inflammation are associated with increases in rapid eye movement (REM) sleep, which is thought to be a prognostic indicator of alcohol relapse. This study was undertaken to test whether blockade of biologically active tumor necrosis factor-alpha (TNF-alpha) normalizes REM sleep in alcohol-dependent adults. METHODS: In a randomized, placebo-controlled, double-blind, crossover trial, 18 abstinent alcohol-dependent male adults received a single dose of etanercept (25 mg) versus placebo in a counterbalanced order. Polysomnographic sleep was measured at baseline and for 3 nights after the acute dose of etanercept or placebo. RESULTS: Compared with placebo, administration of etanercept produced significant decreases in the amount and percentage of REM sleep. Decreases in REM sleep were robust and approached low levels typically found in age-comparable control subjects. Individual differences in biologically active drug as indexed by circulating levels of soluble tumor necrosis factor receptor II negatively correlated with the percentage of REM sleep. CONCLUSIONS: Pharmacologic neutralization of TNF-alpha activity is associated with significant reductions in REM sleep in abstinent alcohol dependent patients. These data suggest that circulating levels of TNF-alpha may have a physiologic role in the regulation of REM sleep in humans. PMID- 19185289 TI - An array of possibilities for pectin. AB - Pectins are a major component of plant cell walls and have numerous roles in plant growth and development. Extracted pectins are widely used as functional food ingredients in products including ice creams, jams, jellies and milk drinks. Although all are based on a galacturonan-rich backbone, pectins are an immensely diverse family of polysaccharides, the functional properties of which are dictated by their fine structures. Understanding the biological roles of pectins and optimizing their industrial usage requires a detailed knowledge of their diversity and spatial and temporal distributions, and microarray technology is a promising tool for high throughput pectin analysis. This article discusses the technical aspects of the production of pectin microarrays and explores their current and potential future uses in the context of recent work in the field. PMID- 19185290 TI - Structure of the lipopolysaccharide core of Vibrio vulnificus type strain 27562. AB - The structure of the lipopolysaccharide core of Vibrio vulnificus type strain 27562 is presented. LPS hydrolysis gave two oligosaccharides, OS-1 and OS-2, as well as lipid A. NMR spectroscopic data corresponded to the presence of one Kdo residue, one beta-glucopyranose, three heptoses, one glyceric acid, one acetate, three PEtN, and one 5,7-diacylamido-3,5,7,9-tetradeoxynonulosonic acid residue (pseudaminic acid, Pse) in OS1. OS2 differed form OS 1 by the absence of glyceric acid, acetate, and Pse residues. Lipid A was analyzed for fatty acid composition and the following fatty acids were found: C14:0, C12:0-3OH, C16:0, C16:1, C14:0 3OH, C18:0, C18:1 in a ratio of 1:3:3:1:2.5:0.6:0.8. PMID- 19185291 TI - Cyclomaltoheptaose mixed esters of anti-inflammatory drugs and short-chain fatty acids and study of their enzymatic hydrolysis in vitro. AB - In an effort to enhance the drug-loading capacity of cyclomaltoheptaose (beta cyclodextrin, betaCD) and to combine the function of anti-inflammatory drugs with short-chain fatty acids (SCFA), ternary esters incorporating seven copies of an anti-inflammatory drug and 14 copies of a SCFA onto a beta-cyclodextrin core were designed and prepared. Acetic, propionic, or butyric esters were introduced at secondary OH groups, and ibuprofen, flurbiprofen, or felbinac was attached to primary OH groups through ester bonds. Heptakis[2,3-di-O-butanoyl-6-O-2-(biphenyl 4-yl)-ethanoyl]-cyclomaltoheptaose was very stable in aqueous and esterase solution. It was hydrolyzed by alpha-amylase (4 units/mL) with t(1/2) value of 18h. The total released amount of biphenyl acetic acid was 38% after 24h when the esterase was added after the alpha-amylase hydrolysis. The present results suggest that these nine betaCD conjugates may release the anti-inflammatory drug in the colonic contents. PMID- 19185292 TI - Design and synthesis of glucose-templated proline-lysine chimera: polyfunctional amino acid chimera with high prolyl cis amide rotamer population. AB - We describe the synthesis of two glucose-templated proline-lysine chimeras (GlcTProLysCs) that differ in the stereochemistry of the hydroxymethyl substituent at the C-5' position of the pyrrolidine ring. The key synthetic steps involve C-glycosylation of an exocyclic glucose-based epoxide with allyltributylstannane, which affords functionalized C-ketosides containing an alpha-hydroxy ester moiety; introduction of an amino group at C-2 through stereoselective reductive amination; and regioselective installation of the azide group at C-6 on the glucose scaffold. Incorporation of these chimeras into the model peptides Ac-GlcTProLysC-NHMe and Ac-GlcTProLysC-OMe demonstrates that the stereochemistry of the hydroxymethyl substituent at the C-5' position has a profound effect on the equilibrium constant of prolyl amide cis/trans isomerization. The equilibrium constant K(c/t) for the peptide mimic Ac GlcTProLysC-NHMe with C-5'(R) stereochemistry was determined to be 3.03+/-0.04, while the K(t/c) for the C-5'(S) diastereoisomer was 0.56+/-0.04 in D(2)O. Temperature coefficient experiments indicate that the origin of these effects is derived from two critical hydrogen bonds involving the C-5' hydroxymethyl substituent: one to the N-terminal amide carbonyl group, and the other to the primary amino group in the glucose moiety. PMID- 19185293 TI - GROMOS96 43a1 performance on the characterization of glycoprotein conformational ensembles through molecular dynamics simulations. AB - Considering the small number of papers assessing the conformational profile of glycoproteins through molecular dynamics (MD) simulations, the current work reports on a systematic analysis of the performance of the GROMOS96 43a1 force field and Lowdin HF/6-31G( * *)-derived atomic charges in the conformational description of glycoproteins. The results substantiate the accuracy of the computational representation of glycoprotein conformational ensembles in aqueous solution based on their agreement to available experimental information, supporting further contributions of computational techniques, mainly MD, in future studies on the characterization of glycoprotein structure and function. PMID- 19185294 TI - Efficient synthesis and protein conjugation of beta-(1-->6)-D-N-acetylglucosamine oligosaccharides from the polysaccharide intercellular adhesin. AB - A wide variety of medically important biofilm forming bacteria produce similar polysaccharide intracellular adhesins (PIAs). The PIA structures consist of partially de-N-acetylated beta-(1-->6)-N-acetylglucosamine polymers. These exopolysaccharides are key components of the bacterial biofilm matrix. Here, we describe the efficient synthesis of PIA oligosaccharides using an acid reversion reaction of N-acetylglucosamine in HF.pyridine. The PIA oligosaccharides produced by this reaction can be purified to homogeneity by size exclusion chromatography. Chemistry was developed to conjugate the PIA oligosaccharides to bovine serum albumin using a new heterobifunctional linker containing a thiol and an N methylhydroxylamine functional group. These glycoconjugates may serve as useful precursors for the development of defined conjugate vaccines against PIA producing bacterial strains. PMID- 19185295 TI - p38 MAPK contributes to CD54 expression and the enhancement of phagocytic activity during macrophage development. AB - p38 is a subfamily of the mitogen-activated protein kinase (MAPK) superfamily with four isoforms. It has been well established that p38 plays a central role in the production of inflammatory molecules and is therefore required for the activation of macrophages in response to inflammatory stimuli. However, little is known about the roles of p38 in macrophage development. The difficulty to get mice deficient in multiple p38 isoforms complicates the study of p38 in macrophage development. With the model of bone marrow-derived murine macrophages and highly selective p38alpha/beta inhibitors SB203580 and SB239063, here we report that macrophage colony-stimulating factor (M-CSF) induces p38 activation during macrophage development. Inhibition of p38 activity showed minor effects on macrophage proliferation or survival, and did not block CD14, F4/80 expression. However, p38 inhibitors resulted in a significant reduction in CD54 expression and impaired phagocytic activity. Taken together, our data suggest that p38 contributes to macrophage development. PMID- 19185296 TI - Atmospheric mercury in Changbai Mountain area, northeastern China I. The seasonal distribution pattern of total gaseous mercury and its potential sources. AB - An intensive field campaign for the measurement of total gaseous mercury (TGM) concentrations in ambient air was conducted in Changbai Mountain area from 5 August 2005 to 5 July 2006 using an automatic atmospheric mercury analyzer (Tekran 2537A), which was the first time TGM was monitored at a remote area in northeastern China. 99% of the hourly TGM concentrations fell between 1.28 and 9.49ngm(-3) with an annual arithmetic mean of 3.58+/-1.78ngm(-3), which was significantly elevated compared to values obtained in remote areas of Europe and North America. Seasonal mean TGM concentrations displayed a descending trend as follows: winter, spring, fall, and summer. Compared to spring/winter, TGM concentrations were lower in the summer/fall but the standard deviation (SD) of TGM levels was higher and indicated a correlation with anthropogenic emissions. TGM concentrations showed seasonal differences with respect to meteorological parameters: TGM levels in spring/winter were most correlated with wind speed, and correlated with solar radiation only in the winter, while TGM levels in summer/fall periods were most correlated with air temperature. There was a strong diurnal variation of seasonal TGM with significantly higher concentrations in daytime/nighttime compared to the early morning. The seasonal diel TGM pattern indicated regional biofuel and coal combustion were the primary mercury sources. PMID- 19185297 TI - Hormonal changes and somatopsychologic manifestations in the first trimester of pregnancy and post partum. AB - OBJECTIVE: To investigate links between psychologic distress and levels of maternal hormones and cortisol during pregnancy and post partum. METHOD: The study population of 308 pregnant or recently delivered women was divided into 4 equal groups, 3 representing each pregnancy trimester and the fourth one representing the first week of the postpartum period. The women were screened for psychological distress and had blood drawn to measure serum levels of estradiol, progesterone, and total cortisol on the same day. RESULTS: The rates of anxiety and depression were higher in the first trimester and the postpartum groups. Depression was correlated with sharp changes in estradiol and progesterone levels; anxiety was correlated with total cortisol level; and there was a correlation between depression and anxiety. CONCLUSION: Sharp changes in maternal hormone levels were correlated with depression; depression was correlated with anxiety; and anxiety was correlated with elevated cortisol levels, which may adversely affect the health of mother and offspring. Measuring maternal hormones may be a diagnostic tool to evaluate psychologic stress and associated risks. PMID- 19185298 TI - Increased carotid intima media thickness is associated with depletion of circulating myeloid dendritic cells in HIV-infected patients on suppressive antiretroviral treatment. PMID- 19185300 TI - A propensity score-based case-control study of renin-angiotensin system gene polymorphisms and diastolic heart failure. AB - Angiotensin II plays an important role in diastolic heart failure (DHF). However, genetic studies of DHF are scarce in the literature. We hypothesized that RAS genes might be the susceptible genes for DHF and conducted a propensity score based case-control study to prove this hypothesis. A total of 666 subjects (285 diagnosed with DHF confirmed by echocardiography and 381 without diastolic dysfunction) were recruited. Genotyped were: the angiotensin-converting enzyme (ACE) gene insertion/deletion polymorphism; the T174M, M235T, G-6A, A-20C, G-152A and G-217A polymorphisms of the angiotensinogen (AGT) gene; and the A1166C polymorphisms of the angiotensin II type I receptor (AT1R) gene. Propensity scores (PS) were used to find patients with and without DHF with equalized characteristics. We also assembled another set of PS matched groups for all characteristics except left ventricular mass (LVM) to detect the genetic association with DHF through the effect of left ventricular hypertrophy. PS matched 210 patients with DHF to 210 without. In a single-locus analysis, the odds ratios (ORs) for DHF were significant with the ACE DD genotype (OR=1.30, 95% CI=1.13-1.49, permuted P=0.003) and the AT1R 1166 CC genotype (OR=2.61, 95% CI=1.52-4.45, permuted P<0.001). Significant gene-gene interaction between the two genes was also detected. However, the ACE gene effect was diminished if LVM was not controlled in the propensity scores. We concluded that genetic variants in the RAS genes may determine individual risk to develop DHF through different pathways. Concomitant presence of ACE DD and AT1R 1166 CC genotypes synergistically increased the predisposition to DHF. PMID- 19185299 TI - Inverse association of erythrocyte n-3 fatty acid levels with inflammatory biomarkers in patients with stable coronary artery disease: The Heart and Soul Study. AB - OBJECTIVE: Dietary intake of polyunsaturated n-3 fatty acids has been associated with a reduced incidence of adverse cardiovascular events. The protective mechanisms involved are not fully understood, but may include anti-inflammatory factors. We sought to investigate the relationship between n-3 fatty acid levels in erythrocyte membranes and markers of systemic inflammation in 992 individuals with stable coronary artery disease. METHODS: Cross-sectional associations of C reactive protein (CRP) and Interleukin-6 (Il-6) with docosahexaenoic acid (DHA) and eicosapentaenoic acid (EHA) were evaluated in multivariable linear regression models adjusted for demographics, cardiovascular risk factors, medication use, exercise capacity, body-mass index, and waist-to-hip ratio. RESULTS: After multivariable adjustment, n-3 fatty acid levels (DHA+EPA) were inversely associated with CRP and IL-6. The inverse association of n-3 fatty acids with CRP and IL-6 was not modified by demographics, body-mass index, smoking, LDL cholesterol, or statin use (p values for interaction>0.1). CONCLUSIONS: In patients with stable coronary artery disease, an independent and inverse association exists between n-3 fatty acid levels and inflammatory biomarkers. These findings suggest that inhibition of systemic inflammation may be a mechanism by which n-3 fatty acids prevent recurrent cardiovascular events. PMID- 19185301 TI - Microvascular degenerative complications are associated with increased aortic stiffness in type 2 diabetic patients. AB - OBJECTIVE: Diabetes is a risk factor for increased arterial stiffness; however, few studies had investigated its associated factors. The aim was to evaluate the correlates of increased arterial stiffness in type 2 diabetes, particularly the relationships with microvascular complications. METHODS: 482 type 2 diabetic patients without peripheral arterial disease were evaluated in a cross-sectional study. Clinical (including tests of cardiovascular dysautonomy), laboratory, ECG, echocardiographic and 24h ambulatory blood pressure monitoring data were obtained. Arterial stiffness was assessed by carotid-femoral (aortic) and carotid radial (peripheral) pulse wave velocity (PWV) measurements. Statistics included multivariate linear and logistic regressions to investigate the independent correlates of increased arterial stiffness. RESULTS: No diabetes-related variable was associated with peripheral arterial stiffness. 148 patients (31%) had increased aortic PWV (>12m/s). On multiple linear regression, retinopathy and nephropathy, besides age, heart rate, 24h pulse pressure, diabetes duration, dyslipidemia and number of antihypertensive drugs in use, were independently associated with aortic PWV. On multivariate logistic regression increased aortic stiffness was associated with retinopathy (odds ratio: 3.83, 95% confidence interval [CI]: 2.24-6.56, p<0.001) and peripheral neuropathy (odds ratio: 1.79, 95%CI: 1.06-3.02, p=0.03) after adjusting for possible confounding variables. Other variables associated with increased aortic stiffness were older age, heart rate, diabetes duration, 24h pulse pressure, dyslipidemia and physical inactivity. CONCLUSIONS: In type 2 diabetic patients, increased central arterial stiffness is associated with the presence of microvascular complications independent of other established determinants of aortic stiffness. PMID- 19185302 TI - Arterial remodeling in response to increased blood flow using a constituent-based model. AB - Previous theoretical models of arterial remodeling in response to changes in blood flow were based on the assumption that material properties of the arterial wall remain unchanged during the remodeling process. According to experimental findings, however, remodeling due to increased flow is accompanied by alteration in the structural properties of elastin, which results in a decrease in its effective elastic stiffness. To account for these effects, we propose a predictive model of arterial remodeling hypothesizing that the variation in mechanical properties of elastin is initiated and driven by the deviation of the intimal shear stress from its baseline value. Geometrical remodeling restores the wall stress distribution as it was under normal flow conditions. A constrained mixture approach is followed. Artery is modeled as a thick-walled cylindrical tube made of non-linear, elastic, anisotropic and incompressible material. Data for a rabbit thoracic aorta have been employed. At the final adapted state, the model predicts a non-monotonic dependence of arterial compliance on the magnitude of flow. This result is in agreement with available experimental data in the literature. PMID- 19185303 TI - Mechanical properties of the porcine growth plate and its three zones from unconfined compression tests. AB - The aim of the study was to determine intrinsic mechanical properties of the complete growth plate and its reserve, proliferative and hypertrophic zones. Growth plate disk samples from newborn swine's ulnae were tested using stress relaxation tests under unconfined compression. The Transversely Isotropic Biphasic Model (TIBPE) derived by [Cohen, B., Lai, W. M., Mow, V. C., 1998. A transversely isotropic biphasic model for unconfined compression of growth plate and chondroepiphysis. Journal of Biomechanical Engineering, 120, pp. 491-496] was used to extract intrinsic mechanical properties using a four-parameter optimization procedure. Significant differences were found for the transverse permeability k(1), the Poisson's ratio in the transverse plane nu(21), the out-of plane Poisson's ratio nu(31) and the out-of-plane Young's modulus E(3) between the reserve zone and the proliferative zone as well as between the reserve zone and the hypertrophic zone. The same trends were obtained for the Young's modulus in the transverse plane E(1), but significant differences were also found between the reserve zone and the complete growth plate. The proliferative and hypertrophic zones are half as stiff as the reserve zone along the compression axis and about three times less stiff than the reserve zone in the transverse plane. These two zones are also three times as permeable as the reserve zone in the radial direction. The mechanical behavior of the newborn porcine distal ulna growth plate is non-uniform along its thickness. The reserve zone, with its greater zonal component at that development stage, has noteworthy effects on the complete growth plate intrinsic mechanical properties. This study provides, for the very first time, an investigation of the intrinsic mechanical properties of the reserve, proliferative and hypertrophic zones of the growth plate. PMID- 19185304 TI - Muscle moment arm and normalized moment contributions as reference data for musculoskeletal elbow and wrist joint models. AB - A geometric musculoskeletal model of the elbow and wrist joints was developed to calculate muscle moment arms throughout elbow flexion/extension, forearm pronation/supination, wrist flexion/extension and radial/ulnar deviation. Model moment arms were verified with data from cadaver specimen studies and geometric models available in the literature. Coefficients of polynomial equations were calculated for all moment arms as functions of joint angle, with special consideration to coupled muscles as a function of two joint angles. Additionally, a "normalized potential moment (NPM)" contribution index for each muscle across the elbow and wrist joints in four degrees-of-freedom was determined using each muscle's normalized physiological cross-sectional area (PCSA) and peak moment arm (MA). We hypothesize that (a) a geometric model of the elbow and wrist joints can represent the major attributes of MA versus joint angle from many literature sources of cadaver and model data and (b) an index can represent each muscle's normalized moment contribution to each degree-of-freedom at the elbow and wrist. We believe these data serve as a simple, yet comprehensive, reference for how the primary 16 muscles across the elbow and wrist contribute to joint moment and overall joint performance. PMID- 19185306 TI - Detailed analysis of petroleum hydrocarbon attenuation in biopiles by high performance liquid chromatography followed by comprehensive two-dimensional gas chromatography. AB - Enhanced bioremediation of petroleum hydrocarbons in two biopiles was quantified by high-performance liquid chromatography (HPLC) followed by comprehensive two dimensional gas chromatography (GCXGC). The attenuation of 34 defined hydrocarbon classes was calculated by HPLC-GCXGC analysis of representative biopile samples at start-up and after 18 weeks of biopile operation. In general, a-cyclic alkanes were most efficiently removed from the biopiles, followed by monoaromatic hydrocarbons. Cycloalkanes and polycyclic aromatic hydrocarbons (PAHs) were more resistant to degradation. A-cyclic biomarkers farnesane, trimethyl-C13, norpristane, pristane and phytane dropped to only about 10% of their initial concentrations. On the other hand, C29-C31 hopane concentrations remained almost unaltered after 18 weeks of biopile operation, confirming their resistance to biodegradation. They are thus reliable indicators to estimate attenuation potential of petroleum hydrocarbons in biopile processed soils. PMID- 19185305 TI - Preparation and binding study of solid-phase microextraction fiber on the basis of ametryn-imprinted polymer: application to the selective extraction of persistent triazine herbicides in tap water, rice, maize and onion. AB - A monolithic ametryn molecular-imprinted polymer based on a simple polymerization method was fabricated for use as new solid-phase microextraction (SPME) fiber, which can be coupled with GC and GC/MS for selective extraction and analysis of triazine herbicides. Methacrylic acid (MAA), ethylene glycol dimethacrylate (EDMA) and ametryn bear role of functional monomer, cross-linker and template, respectively. In the optimized conditions the fabricated fiber showed better molecular recognition abilities for methylthiotriazine herbicides than chloro triazine herbicides. By use of bi-Langmuir isotherm model the evaluated equilibrium constants for ametryn were 0.01 and 890.69 microM(-1), and the numbers of binding sites were 129.98 and 5.82 nmol g(-1), respectively. The high extraction efficiency was obtained for ametryn, prometryn, terbutryn, atrazine, simazine, propazine, and cyanazine, yielding the detection limits of 14, 28, 45, 56, 85, 95 and 74 ng mL(-1), respectively by GC with flame ionization detection. The reliability of the prepared fiber for extraction of ametryn and other analogues in real samples has been investigated and proved by using spiked samples such as tap water, rice, maize, and onion. PMID- 19185307 TI - Molecularly imprinted layer-coated silica nanoparticles toward highly selective separation of active diosgenin from Dioscorea nipponica Makino. AB - This paper reports the molecularly imprinted layer-coated silica nanoparticles toward highly selective separation of active diosgenin (DG) from the crude extracts of Dioscorea nipponica Makino (DNM). It has been demonstrated that DG templates were efficiently imprinted into the silica layer by the use of thermally cleavable urethane bonds between DG and 3 isocyanatopropyltriethoxysilane (IPTS), which was synchronously coated onto the surface of silica mother nanoparticles through a seed-directing surface condensation reaction between DG-IPTS and tetraethoxysilicane (TEOS). After removal of templates by simple thermal cleavage reaction, a high density of recognition sites of DG were created in the silica-coating layer. Afterwards, the DG-imprinted silica nanoparticles were evaluated by rebinding experiments and showed a higher selectivity and affinity to DG than commercial silica. When the imprinted particles were used as solid-phase extraction (SPE) sorbents, the recovery yield of DG was up to 90% by one-step extraction from the hydrolysate of DNM, and the purity of DG was larger than 98% by HPLC analysis. These results reported herein provide the possibility for the highly selective separation and purification of active DG from DNM by the molecular imprinting modification at the surface of common silica adsorbents. PMID- 19185308 TI - Total concentration analysis of polycylic aromatic hydrocarbons in aqueous samples with high suspended particulate matter content. AB - The European water framework directive (WFD) requires priority pollutants to be measured in the whole water sample and not only in the dissolved phase. However, it does not give clear definitions on how to achieve this. To overcome this limitation, a new methodology of sample preparation procedure for the analysis of polycyclic aromatic hydrocarbons on the basis of extraction disks is introduced here. The automatable procedure includes a "one-step" extraction of the analytes both dissolved in the liquid phase of the sample and sorbed to suspended particulate matter. The latter is extracted concurrently with the solid-phase extraction (SPE) material within the elution step of the procedure. Separation, identification, and quantification of the analytes is performed by GC-MS. Results from surface water samples spiked with certified sediment up to 1000 mg/l are presented in this work and compared with results derived from liquid-liquid extraction (LLE). Most measured values are within or at least near certified uncertainty limits of the sediment. The SPE disk method shows much higher recoveries and better precision (relative standard deviations between 2% and 11%) than the standard LLE method. For all substances under investigation, the limits of quantification achieved range between 0.001 and 0.005 microg/l. PMID- 19185309 TI - Statistical optimization of the silylation reaction of a mercaptosilane with silanol groups on the surface of silica gel. AB - Thiol-modified silica is often used as an intermediate product for further synthesis of modified stationary phases for chromatography or purification processes. Different conditions were used to synthesize such thiol-modified particles, but systematic optimizations remained scarce. In this study the reaction conditions for the synthesis of mercaptopropyl-modified silica were optimized. The general synthetic method consists in slurrying the silica gel in toluene before adding 3-mercaptopropyldimethoxymethylsilane together with a tertiary amine as catalyst (here dimethylaminopyridine). Reaction time and temperature were optimized using a full factorial design of experiment (DoE) from 3 to 25h with temperature varying between 45 and 105 degrees C. The surface coverage of the silica with mercaptopropyl-groups was analyzed by two different ways (elemental analysis and chemical surface reaction with 2,2'-dipyridyl disulfide followed by HPLC-UV analysis of stoichiometrically liberated pyridyl-2 thione). We obtained a three-dimensional (3D) plot of the surface coverage as a function of reaction time and temperature. The arch-shaped hyperplane allowed us to determine an optimum with regard to time and temperature, which yields to the highest surface coverage possible. We also verified that the increase of the surface coverage does not lead to a decrease of the stability of the surface modification by subjecting the gels to treatment with high temperature and acidic conditions. The stability was monitored by different chromatographic methods. Moreover, (29)Si cross-polarization-magic angle spinning (CP-MAS) NMR spectra of materials prepared by different conditions allowed to confirm that the Si species on the surface were essentially the same, while there was only a minute difference in signal intensities for the individual Si species for materials obtained by distinct temperatures. PMID- 19185310 TI - Equilibrium morphologies of nonionic lipid-nanoparticle mixtures in water: a self consistent mean-field prediction. AB - We model an aqueous system which comprises a mixture of lipid molecules and hydrophobic nanoparticles, by means of a self-consistent mean-field theory (SCMFT). Lipids are modeled as nonlinear/branched copolymers with a single hydrophilic head group and a double hydrophobic tail, whereas nanoparticles are modeled as hard-spheres of a particular size. The mixture of lipids and nanoparticles leads to a formation of core-shell micellar structures where the hydrophobic nanoparticles and lipid tails form the core of the micelle, and the hydrophilic lipid head groups form the shell. Different micellar morphologies are found depending on the total concentration of lipid molecules and nanoparticles, as well as the relative size of nanoparticles. There exist three distinct equilibrium morphologies of lipid-nanoparticle micelles: circular micelles (CM), ellipsoidal micelles (EM), and bilayer/lamellar structures (BL). We observed some smooth morphological transitions and phase coexistences by evaluating the excess free energy of micelles. PMID- 19185311 TI - Morphology of anisotropic chains in a magneto-rheological fluid during aggregation and disaggregation processes. AB - We study the morphology of the chain-like aggregates formed when a external constant and uniaxial magnetic field is applied to a magneto-rheological (MR) fluid. In order to characterize the conformation of the aggregates, we study the evolution of various fractal dimensions during aggregation and disaggregation processes (i.e., when the applied field is switched on and off), using video microscopy and image analysis. Experiments have been performed by varying the values of two external parameters: the magnetic field amplitude and particle concentration. We found that the box-counting dimension, related with how the aggregates occupy the surrounding space, depends on the ratio R(1)/R(0). During the first stage of the disaggregation process, when the particles are moving by Brownian motion inside the aggregate, Family-Vicsek scaling function is verified. PMID- 19185312 TI - Effect of buffers on aqueous solute-exclusion zones around ion-exchange resins. AB - Interaction between charged surfaces in aqueous solution is a fundamental feature of colloid science. Theoretically, surface potential falls to half its value at a distance equal to a Debye length, which is typically on the order of tens to hundreds of nanometers. This potential prevents colloids from aggregating. On the other hand, long-range surface effects have been frequently reported. Here we report additional long-range effects. We find that charged latex particles in buffer solutions are uniformly excluded from several-hundred-micron-thick shells surrounding ion-exchange beads. Exclusion is observed whether the beads are charged similarly or oppositely to the particles. Hence, electrostatic interactions between bead and microsphere do not cause particle exclusion. Rather, exclusion may be the consequence of water molecules re-orienting to produce a more ordered structure, which then excludes the particles. PMID- 19185313 TI - Mixed adsorption layers of 0.1 M tert-butanol-tetramethylthiourea at the interface of Hg/aqueous perchlorate solutions. AB - The electrosorption behavior of tetramethylthiourea (TMTU) on a mercury electrode from 1.0, 0.5, and 0.1 M NaClO(4) solutions containing 0.1 M tert-butanol (TB) is described by means of adsorption isotherm constants. The adsorption parameters for the double layer were calculated from a double-layer differential capacity measurement extrapolated to zero frequency. The values of the relative surface excess increase along with the concentration of NaClO(4). It seems that the adsorption of TMTU is determined by an interaction between adsorbed molecules. The presence of TB in the adsorption layer distinctly influences the parameters of the inner layer. PMID- 19185316 TI - Positive screening results on the modified checklist for autism in toddlers: implications for very preterm populations. PMID- 19185315 TI - Imaging myocardial scar and arrhythmic risk prediction--a role for the electrocardiogram? AB - Risk stratification for sudden cardiac death (SCD) has become increasingly important to identify candidates for implantable cardioverter-defibrillators (ICDs). Existing clinical guidelines to identify patients for ICDs focus on reduced left ventricular ejection fraction (LVEF); however, the average annual rate of appropriate ICD shocks is only 5.1% in this select group (LVEF 70, the odds for screening positive were increased 13-fold for those with an MDI of <55 and more than 4-fold for those with an MDI of 55 to 69. CONCLUSIONS: Major motor, cognitive, visual, and hearing impairments appear to account for more than half of the positive M-CHAT screens in extremely low gestational age newborns. Even after those with such impairments were eliminated, 10% of children--nearly double the expected rate--screened positive. PMID- 19185318 TI - Amelioration of white-matter lesions in a patient with Fabry disease. AB - We report on a 27-year-old man with Fabry disease who had widespread white-matter lesions (WMLs) despite the absence of renal or cardiac manifestations. Genomic analysis revealed a novel mutation: a GAT deletion at nucleotide position 234-236 in exon 5 of the coding region. After 12 months of enzyme replacement therapy (ERT), most of the WMLs had disappeared. Cell counts and protein levels in the cerebrospinal fluid also decreased. These findings suggest that ERT may play a role in the recovery of WMLs. PMID- 19185319 TI - Differentiating ischemic stroke subtypes: risk factors and secondary prevention. AB - Ischemic strokes account for 87% of all strokes in the US. Patients who suffer an initial ischemic stroke or transient ischemic attack are at risk for recurrent strokes, as well as ischemic events in the coronary and peripheral vasculatures. Lifestyle modifications and pharmacological and surgical interventions are effective in reducing the risk of recurrent events in all stroke patients, and evidence-based guidelines for secondary stroke prevention are available. However, increasing evidence indicates that risk factors may be differentially associated with specific ischemic stroke subtypes. In the future, exploiting these differential associations may facilitate secondary stroke prevention and, therefore, improve patient outcomes. PMID- 19185320 TI - Reduced neurogenesis after suppressed inflammation by minocycline in transient cerebral ischemia in rat. AB - Recently, the beneficial role of minocycline on endogenous neurogenesis after cerebral ischemia has been contradicted by many reports. We examined whether minocycline influences post-ischemic neurogenesis in the subventricular zone. Adult male Sprague-Dawley rats were subjected to focal cerebral ischemia for 2 h, and divided into a minocycline-treated (90 mg/Kg on reperfusion and 45 mg/Kg daily for maintenance) and a saline-treated group. Bromodeoxyuridine was injected to determine levels of cell proliferation. Inflammation was assessed by counting polymorphonuclear cell and activated microglia and by measuring myeloperoxidase activity. Endogenous neurogenesis was quantified by immunohistochemical staining and functional outcome was measured by infarct size and behavioral tests. Minocycline treatment decreased inflammation on 1st and 4th days after ischemia. BrdU-positive cells on 7th day (saline vs. minocycline: 602.80+/-146.96 vs. 399.40+/-109.69) and the number of double labeling cells of BrdU/NeuN on 7th day (13.00+/-4.36 vs. 6.40+/-2.07) and BrdU/DCx on 4th day (17.00+/-5.00 vs. 7.50+/ 1.91) were significantly decreased in minocycline-treated rats. Infarct size and behavioral tests were not different. Our results indicate that minocycline may reduce immediate post-ischemic neurogenesis despite adequately suppressed inflammation. PMID- 19185321 TI - The relationship between premature ejaculation and hyperthyroidism. AB - PURPOSE: We determine the prevalence of premature ejaculation in patients with hyperthyroidism and observed intravaginal ejaculation latency time alterations before and after hyperthyroidism treatment. MATERIALS AND METHODS: Between January 2004 and June 2007, 49 patients with hyperthyroidism and no history of hyperthyroidism treatment were enrolled in the study. After obtaining a detailed sexual anamnesis an erectile function questionnaire was completed and a patient self-reported outcome measure of difficult control over ejaculation was examined. We assessed stopwatch measurements of intravaginal ejaculation latency time performed by the patient or partner. Patient anxiety status was also evaluated. Changes in the mentioned measurements induced by hyperthyroidism treatment were examined 8 weeks after the achievement of euthyroidism. RESULTS: In the 43 eligible patients mean +/- SD age was 48.0 +/- 8.8 years. Premature ejaculation was observed in 31 of the 43 patients (72.1%). Mean intravaginal ejaculation latency time in patients with hyperthyroidism was 72.8 +/- 83.3 seconds. Of the 43 patients 30 (69.8%) were considered to have definite premature ejaculation according to stopwatch measurements. In patients with hyperthyroidism who had definite premature ejaculation anxiety scores were determined to be higher. A positive correlation was noted between serum thyroid stimulating hormone and intravaginal ejaculation latency time in the patients. In 24 patients who completed the followup visits we noted statistically significant improvement in intravaginal ejaculation latency time after the achievement of euthyroidism. CONCLUSIONS: Excess thyroid hormone and premature ejaculation are clinically interrelated conditions. Hyperthyroidism should be considered a novel and reversible etiological risk factor for premature ejaculation. PMID- 19185322 TI - A panel of angiogenic factors for early bladder cancer detection: enzyme immunoassay and Western blot. AB - PURPOSE: Angiogenesis is tightly regulated by a large number of pro-angiogenic factors, including vascular endothelial growth factor, basic fibroblast growth factor, hepatocyte growth factor and angiogenin. We adapted and evaluated the measurement of these factors using enzyme-linked immunosorbent assay and compared the results with Western blot and voided urine cytology. MATERIALS AND METHODS: This study included 240 patients diagnosed with bladder carcinoma, 108 with benign bladder lesions and 110 healthy individuals who served as controls. All participants underwent serological schistosomiasis antibody assay in serum, urine cytology and estimation of angiogenic factors in voided urine. RESULTS: Intra assay and interassay CVs of the investigated markers were 10.3 to 12.3 and 10 to 13.7, respectively. The recovery rate of the added angiogenic factor to the urine pool was 98% to 103%, 97% to 103%, 98% to 104% and 97% to 100% for vascular endothelial growth factor, basic fibroblast growth factor, angiogenin and hepatocyte growth factor, respectively. The concordance rate with Western blot was 97.5%. The levels and positive rates of urinary angiogenic markers and urine cytology were significantly higher in the malignant group than in the benign and healthy groups. Basic fibroblast growth factor increased significantly in bladder squamous cell carcinoma cases. Moreover, basic fibroblast growth factor and hepatocyte growth factor significantly correlated with tumor grade. Angiogenic markers showed significant association with clinical stage. CONCLUSIONS: Quantitative measurement of urinary angiogenic factors in voided urine samples by enzyme-linked immunosorbent assay was reliable. The sensitivity of basic fibroblast growth factor and hepatocyte growth factor was superior to that of the other investigated markers and of cytology in low grade and early stage cases, suggesting their convenience as sensitive, noninvasive diagnostic and screening tools for bladder cancer. PMID- 19185323 TI - Bacterial flora and antibiotic resistance from eggs of green turtles Chelonia mydas: an indication of polluted effluents. AB - Sea turtles migrate to various habitats where they can be exposed to different pollutants. Bacteria were collected from turtle eggs and their resistance to antibiotics was used as pollutant bio-indicators of contaminated effluents. Eggs were collected randomly from turtles when they were laying their eggs. A total of 90 eggs were collected and placed into sterile plastic bags (3 eggs/turtle) during June-December of 2003. The bacteria located in the eggshell, albumen and yolk were examined, and 42% of the eggs were contaminated with 10 genera of bacteria. Pseudomonas spp. were the most frequent isolates. The albumen was found to be the part of the egg to be the least contaminated by bacterial infection. Bacterial isolates tested with 14 antibiotics showed variations in resistance. Resistance to ampicillin was the highest. The presence of antibiotic resistant bacteria in eggs indicates that the green turtle populations were subjected to polluted effluents during some of their migratory routes and feeding habitats. Scanning electron microscopy revealed that Salmonella typhimurium penetrated all eggshell layers. PMID- 19185324 TI - A case study of in situ oil contamination in a mangrove swamp (Rio De Janeiro, Brazil). AB - Mangroves are sensitive ecosystems of prominent ecological value that lamentably have lost much of their areas across the world. The vulnerability of mangroves grown in proximity to cities requires the development of new technologies for the remediation of acute oil spills and chronic contaminations. Studies on oil remediation are usually performed with in vitro microcosms whereas in situ experiments are rare. The aim of this work was to evaluate oil degradation on mangrove ecosystems using in situ microcosms seeded with an indigenous hydrocarbonoclastic bacterial consortium (HBC). Although the potential degradation of oil through HBC has been reported, their seeding directly on the sediment did not stimulate oil degradation during the experimental period. This is probably due to the availability of carbon sources that are easier to degrade than petroleum hydrocarbons. Our results emphasize the fragility of mangrove ecosystems during accidental oil spills and also the need for more efficient technologies for their remediation. PMID- 19185325 TI - Impact of the salt stress on the photosynthetic carbon flux and 13C-label distribution within floridoside and digeneaside in Solieria chordalis. AB - The flux of photosynthetic carbon used in the synthesis of low-molecular weight carbohydrates (digeneaside and floridoside) was investigated by (13)C and (1)H NMR spectroscopy in samples of the red seaweed, Solieria chordalis, incubated at different salinities (22, 34 and 50psu). Carbohydrates were labelled, by pulse chase, with the stable isotope (13)C from NaH(13)CO(3). In vivo NMR analyses carried out with a cryogenic probe optimised for (13)C detection were performed directly on the living algal tissues to evidence the labelling of the carbohydrates with neither preliminary extraction nor purification step(s). The isotopic enrichment of each compound was determined by high-resolution (1)H and (13)C NMR spectroscopy. These analyses evidenced different orientations of the flux of the photosynthetic carbon in the algae according to the salt stress. At normal and low salinities, the photosynthetic carbon flux was responsible of 70% and 67% of the floridoside synthetized during the pulse period, respectively, whereas it was only of 30% in the thalli exposed to the high salinity, meaning a biosynthesis of high floridoside amount from endogen source leading to the osmotic regulation. Under normal and hyper-osmotic conditions, the stock of floridoside was used for cellular needs during the chase period, whereas it was not under hypo-osmotic conditions. The characterization of isotopomers composition of floridoside and digeneaside and the analysis of adjacent (13)C labelling gives much more details on the effects of salinity on the metabolic pathways leading to the synthesis or the degradation of these molecules. High turnover of floridoside was evidenced at normal salinity during the chase period and products issued from the degradation of floridoside would not be used for the novo biosynthesis. That suggests that synthesis and degradation of floridoside may be realized in two different cellular compartments. The presence of more numerous (13)C-(13)C blocks in the carbon skeleton of the molecules from the up salt stressed thalli than in those from no salt stressed algae, concomitant with a slower degree of isotopic enrichment of the molecule, provided evidence that the two metabolic pathways (endogen and photosynthetic) may not share the precursor molecules involved in the floridoside synthesis and that these two routes may be totally separate until the constitution of floridoside molecule. PMID- 19185326 TI - Straightforward and rapid determination of sulfadoxine and sulfamethoxazole in capillary blood on sampling paper with liquid chromatography and UV detection. AB - A method for the determination of sulfadoxine and sulfamethoxazole in capillary blood on sampling paper has been developed and validated. The method is straightforward with minimal sample preparation, and is suitable for rural settings. Separation of sulfadoxine, sulfamethoxazole and internal standard was performed using a Purospher STAR RP-18 endcapped LC column (150x4.6mm) with a mobile phase consisting of acetonitrile:sodium acetate buffer pH 5.2, I=0.1 (1:2, v/v). For sulfadoxine, the within-day precision was 5.3% at 15micromol/l and 3.7% at 600micromol/l, while for sulfamethoxazole it was 5.7% at 15micromol/l and 3.8% at 600micromol/l. The lower limit of quantification was determined to 5micromol/l and precision was 5.5% and 5.0% for sulfadoxine and sulfamethoxazole, respectively. PMID- 19185327 TI - Asymmetric total synthesis and antiproliferative activity of goniothalamin oxide isomers. AB - Goniothalamin oxide (1) is a styryl lactone which was isolated from bark and leaves of several Goniothalamus species. This natural product has some interesting biological properties such as larvicidal and tripanocidal activities. However, no studies on the antiproliferative profile of goniothalamin oxide (1) and its stereoisomers have been reported yet. Here, goniothalamin epoxide (1), isogoniothalamin epoxide (2) and their enantiomers were prepared via epoxidation of (R)-and (S)-goniothalamin (4). A 3:2 molar ratio in favor of goniothalamin oxide (1) and ent-1 was observed from (R)- and (S)-4, respectively, when 3 chloroperbenzoic acid (mCPBA) was employed while an increase to 6:1 molar ratio was achieved with (S,S)-Jacobsen's catalyst. Antiproliferative activity of these epoxides revealed that ent-isogoniothalamin oxide (ent-2) was the most active against the eight cancer cell lines studied. These results indicate that 6S, 7R and 8R absolute configurations are beneficial for the activity of these epoxides. PMID- 19185328 TI - Significance of treated agrowaste residue and autochthonous inoculates (Arbuscular mycorrhizal fungi and Bacillus cereus) on bacterial community structure and phytoextraction to remediate soils contaminated with heavy metals. AB - In this study, we analyzed the impact of treatments such as Aspergillus niger treated sugar beet waste (SB), PO4(3-) fertilization and autochthonous inoculants [arbuscular mycorrhizal (AM) fungi and Bacillus cereus], on the bacterial community structure in a soils contaminated with heavy metals as well as, the effectiveness on plant growth (Trifolium repens). The inoculation with AM fungi in SB amended soil, increased plant growth similarly to PO4(3-) addition, and both treatments matched in P acquisition but bacterial biodiversity estimated by denaturing gradient gel electrophoresis of amplified 16S rDNA sequences, was more stimulated by the presence of the AM fungus than by PO4(3-) fertilization. The SB amendment plus AM inoculation increased the microbial diversity by 233% and also changed (by 215%) the structure of the bacterial community. The microbial inoculants and amendment used favoured plant growth and the phytoextraction process and concomitantly modified bacterial community in the rhizosphere; thus they can be used for remediation. Therefore, the understanding of such microbial ecological aspects is important for phytoremediation and the recovery of contaminated soils. PMID- 19185329 TI - WITHDRAWN: Book review. AB - This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 19185330 TI - Lead (II) detection and contamination routes in environmental sources, cookware and home-prepared foods from Zimatlan, Oaxaca, Mexico. AB - An interdisciplinary investigation, involving environmental geochemists, epidemiologists, nurses, and anthropologists, was undertaken to determine the contamination source and pathway of an on-going outbreak of lead poisoning among migrants originating from Zimatlan, Oaxaca, Mexico and living in Seaside, California, and among their US-born children. An initial investigation in Seaside identified grasshopper foodstuff ("chapulines") imported from Mexico and consumed as snacks, as containing alarmingly high lead concentrations (up to 2300 mg/kg). The focus in the present work concentrates on the Oaxacan area of origin of the problem in Mexico, and two potential sources of contamination were investigated: wind-borne dusts from existing mine residues as potential contaminants of soil, plant, and fauna; and food preparation practices using lead-glazed ceramic cookware. Over a three year period, sampling was conducted in Oaxaca using community-level sampling and also targeted sampling with families of cases with lead poisoning in California. In addition to fresh field chapulines, we analyzed for total lead: soil, water, mine residues, and plant materials, both from areas adjacent to or at an abandoned waste site containing mine tailings, and from fields where chapulines are collected; foodstuffs gathered in community markets or in a food transport business; and foodstuffs and cookware gathered from relatives of case families in California. Also, selected new and used lead-glazed clay cookware was extracted for lead, using 0.02 M citric acid and with 4% acetic acid. The results indicated significant presence of lead in mine wastes, in specific foodstuffs, and in glazed cookware, but no extensive soil contamination was identified. In-situ experiments demonstrated that lead incorporation in food is made very efficient through grinding of spices in glazed cookware, with the combination of a harsh mechanical action and the frequent presence of acidic lime juice, but without heating, resulting in high but variable levels of contamination. PMID- 19185331 TI - Particle and gaseous emissions from compressed natural gas and ultralow sulphur diesel-fuelled buses at four steady engine loads. AB - Exhaust emissions from thirteen compressed natural gas (CNG) and nine ultralow sulphur diesel in-service transport buses were monitored on a chassis dynamometer. Measurements were carried out at idle and at three steady engine loads of 25%, 50% and 100% of maximum power at a fixed speed of 60 km h(-1). Emission factors were estimated for particle mass and number, carbon dioxide and oxides of nitrogen for two types of CNG buses (Scania and MAN, compatible with Euro 2 and 3 emission standards, respectively) and two types of diesel buses (Volvo Pre-Euro/Euro1 and Mercedez OC500 Euro3). All emission factors increased with load. The median particle mass emission factor for the CNG buses was less than 1% of that from the diesel buses at all loads. However, the particle number emission factors did not show a statistically significant difference between buses operating on the two types of fuel. In this paper, for the very first time, particle number emission factors are presented at four steady state engine loads for CNG buses. Median values ranged from the order of 10(12) particles min(-)(1) at idle to 10(15) particles km(-)(1) at full power. Most of the particles observed in the CNG emissions were in the nanoparticle size range and likely to be composed of volatile organic compounds The CO2 emission factors were about 20% to 30% greater for the diesel buses over the CNG buses, while the oxides of nitrogen emission factors did not show any difference due to the large variation between buses. PMID- 19185332 TI - Coal is a potential source of naphthenic acids in groundwater. AB - Naphthenic acids, with the general formula C(n)H(2n+Z)O(2), are found in conventional petroleums and oil sands ores. These acids are toxic to aquatic life, so their discharge from petroleum processing into receiving waters must be avoided. In a previous study, naphthenic acids were putatively identified in groundwaters from two domestic wells that were distant from petroleum sources. However, coal deposits were near these wells. In this study, waters from the two wells were extracted and analyzed by electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry to unequivocally confirm the presence of naphthenic acids and other organic acids. In addition, distilled water was percolated through three crushed coal samples and the leachates were shown to contain a variety of organic acids, including naphthenic acids. These results clearly demonstrate that coal is a source of naphthenic acids and that the naphthenic acids can leach into groundwaters. Thus, the presence of naphthenic acids in waters cannot be solely attributed to petroleum or petroleum industry activities. PMID- 19185333 TI - Human impacts on urban subsurface environments. PMID- 19185334 TI - Transfer, distribution and bioaccumulation of microcystins in the aquatic food web in Lake Taihu, China, with potential risks to human health. AB - In this paper, accumulation and distribution of microcystins (MCs) was examined monthly in six species of fish with different trophic levels in Meiliang Bay, Lake Taihu, China, from June to November 2005. Microcystins were analyzed by liquid chromatography electrospray ionization mass spectrometry (LC-ESI-MS). Average recoveries of spiked fish samples were 67.7% for MC-RR, 85.3% for MC-YR, and 88.6% for MC-LR. The MCs (MC-RR+MC-YR+MC-LR) concentration in liver and gut content was highest in phytoplanktivorous fish, followed by omnivorous fish, and was lowest in carnivorous fish; while MCs concentration in muscle was highest in omnivorous fish, followed by phytoplanktivorous fish, and was lowest in carnivorous fish. This is the first study reporting MCs accumulation in the gonad of fish in field. The main uptake of MC-YR in fish seems to be through the gills from the dissolved MCs. The WHO limit for tolerable daily intake was exceeded only in common carp muscle. PMID- 19185335 TI - Predicting dissolved inorganic nitrogen leaching in European forests using two independent databases. AB - Regional-scale databases can be particularly useful for identifying relationships between dissolved inorganic nitrogen (N) leaching in forests and environmental drivers, which in turn allow an assessment of the risk of ecosystem damage, such as forest acidification and eutrophication of downstream water bodies. However, detecting the 'signal' of a significant correlate to N leaching against a background of wide variability in other factors requires a large number of sites, and the validation of models developed requires a similarly large number of independent sites. Here we use two large and fully independent databases of forest ecosystems across Europe to develop and validate indicators of N saturation and leaching. One database was used for model development and the other for validating these models. Among 35 variables considered, the most significant indicators of N leaching in the model development database were: the flux of dissolved inorganic N in deposition, mean annual temperature, mean altitude, the site drainage (plot vs catchment), needle- and litter-N concentration, organic horizon C:N ratio, and subsoil pH. Altitude was not a consistent predictor (it was significant in the development database but not in the validation database), and needle and litter N concentration, plot vs catchment, and subsoil pH all showed high intercorrelation with N deposition and so were not significant in models already including N deposition. The most consistent and useful indicators of N leaching were throughfall N deposition, organic horizon C:N ratio and mean annual temperature. Sites receiving low levels of N deposition (<8 kg N ha(-1) y(-1)) showed very low output fluxes of N and were simulated separately from more polluted forests. In general, the models successfully predicted N leaching (mean of +/-5 kg N ha(-1) y(-1) between observed and predicted) from forests at early to intermediate stages of nitrogen saturation but not from nitrogen-saturated sites. Thus, simple relationships developed from combining (1) external drivers (deposition, temperature) and (2) site conditions (nitrogen status of soils) can successfully estimate nitrogen leaching from forests that have not yet been highly damaged by N deposition. PMID- 19185336 TI - Adenosine inhibition of adenosine diphosphate and thrombin-induced monocyte platelet aggregates in cardiac syndrome X. AB - INTRODUCTION: We previously showed that platelet reactivity at rest is increased in patients with cardiac syndrome X (CSX), but that exercise reduces platelet reactivity in these patients. Adenosine was suggested to be involved in this phenomenon. In this study we investigated the effect of adenosine on adenosine diphosphate (ADP) and thrombin-induced platelet reactivity in CSX patients. MATERIALS AND METHODS: We studied 15 CSX patients and a control group of 15 healthy subjects. Formation of monocyte-platelet (MONO-PLT) aggregates in vitro was assessed by flow cytometry: 1) at baseline; 2) after ADP (10(-7) M) stimulation alone; 3) after ADP stimulation in presence of adenosine (10(-5) M); 4) after thrombin (10(-11) M) stimulation alone; 5) after thrombin stimulation in presence of adenosine. RESULTS: In non stimulated samples there were no relevant differences between the two groups in cytometry variables. Compared to controls, ADP induced a higher increase in MONO-PLT aggregates in CSX patients (P < 0.01), which was significantly inhibited by adenosine (P < 0.01). Thrombin also induced a greater increase in MONO-PLT aggregates in CSX patients (P < 0.001), which was also significantly blunted by adenosine. Similar trends were observed for platelet CD41 (glycoprotein IIb-IIIa) receptor and for monocyte receptors CD142 ad CD162 in MONO-PLT aggregates. CONCLUSIONS: In CSX patients platelet reactivity is increased at rest, compared to healthy controls. Pre-incubation with adenosine reduces the agonist-induced platelet hyper-reactivity in these patients, suggesting that adenosine may be involved in the reduction of platelet reactivity observed in CSX patients after exercise in our previous study. PMID- 19185337 TI - Elongator controls the migration and differentiation of cortical neurons through acetylation of alpha-tubulin. AB - The generation of cortical projection neurons relies on the coordination of radial migration with branching. Here, we report that the multisubunit histone acetyltransferase Elongator complex, which contributes to transcript elongation, also regulates the maturation of projection neurons. Indeed, silencing of its scaffold (Elp1) or catalytic subunit (Elp3) cell-autonomously delays the migration and impairs the branching of projection neurons. Strikingly, neurons defective in Elongator show reduced levels of acetylated alpha-tubulin. Reduction of alpha-tubulin acetylation via expression of a nonacetylatable alpha-tubulin mutant leads to comparable defects in cortical neurons and suggests that alpha tubulin is a target of Elp3. This is further supported by the demonstration that Elp3 promotes acetylation and counteracts HDAC6-mediated deacetylation of this substrate in vitro. Our results uncover alpha-tubulin as a target of the Elongator complex and suggest that a tight regulation of its acetylation underlies the maturation of cortical projection neurons. PMID- 19185338 TI - Effects of caffeine, cumulus cell removal and aging on polyspermy and embryo development on in vitro matured and fertilized ovine oocytes. AB - The objectives of these studies were to determine the effects of cumulus cell removal and caffeine treatment on the development of in vitro matured ovine oocytes aged in vitro until until fertilization. Oocytes were denuded (DO) at 24h post-onset of maturation (hpm), control cumulus oocyte complexes (COC's) and DO groups were fertilized at 24 hpm or returned to culture in the presence or absence of 10mM caffeine and fertilized at 30 hpm. Removal of cumulus cells and aging both increased polyspermy, caffeine reduced this increase, however, with the exception of DO's (30 hpm) vs. COC's (24 hpm) the differences were not statistically significant. Aging significantly decreased cleavage between COC groups at 24 hpm and 30 hpm and caffeine did not affect this (68.4%, 73.4%, 74.0% respectively). In contrast, the frequency of cleavage was significantly reduced in the DO (24 hpm) group as compared to COC controls (45.6% vs. 68.4% (P<0.05)), however, cleavage increased in the DO group on aging (73.4%) and this was not affected by caffeine (73.0%). The percentage of COC's and DO's developing to the blastocyst stage significantly decreased on aging, caffeine treatment of DO's prevented this (31.3%, 12.7% and 29.4% respectively (P<0.05)) but had no effect on COC's (4.2% vs. 3.9%). Total cell numbers in blastocysts were not statistically different (92.4+/-5.2, 84.7+/-3.7 and 80.4+/-5.8 (P>0.05)). In summary caffeine treatment of aged COC's had no significant effect on the frequency of development, however, in aged DO's caffeine treatment statistically increased development to blastocyst and lowered the frequency of polyspermy. PMID- 19185339 TI - The relationship between consecutive pregnancies in Thoroughbred mares. Does the location of one pregnancy affect the location of the next, is this affected by mare age and foal heat to conception interval or related to pregnancy success. AB - Delayed uterine involution is a major cause of early reproductive failure in mares. Involution is affected by mare age, and foaling to covering interval. Involution rates vary between the previously non-gravid horn (PNGH), which recovers the quicker, and the previously gravid horn (PGH). Location of a pregnancy and its likely success may, therefore, be affected by its location relative to the previous pregnancy. This study aimed to determine: (i) the location of concepti in consecutive pregnancies; (ii) whether this varies with mare age or foaling to conception interval; (iii) whether location in relation to the previous pregnancy affects success. 1383 Thoroughbred mares were monitored by ultrasonic scanning during oestrus and early pregnancy. Significantly (p<0.01) more pregnancies were located in the PNGH (79.2%) than the PGH (20.8%). The number of pregnancies in PGH significantly increased with mare age (p<0.01) and foaling to conception interval (p<0.05). Significantly (p<0.001) more pregnancies located in the PGH (16.5%) failed, than those in the PNGH (4.6%). It can be concluded that most pregnancies locate in the PNGH where their chances of success are greatest. The larger number of pregnancies locating in the PGH in older mares and those with shorter foaling to conception intervals may in part account for the higher conceptus mortality rates in such mares. Hence breeding older mares on alternate years and maximising foaling to conception interval may improve reproductive success. Alternatively termination of pregnancies located in the PGH followed by timely recovering may be justifiable as might ET in older mares covered close to foaling. PMID- 19185340 TI - Efficiency of estrous synchronization in tropical sheep by combining short interval cloprostenol-based protocols and "male effect". AB - This study assessed the efficacy of a protocol combining short-interval cloprostenol-based protocols and "male effect" for estrous synchronization in hair sheep. In Experiment 1, 24 ewes were randomly assigned to three groups (n=8) and treated with cloprostenol on Days 3, 5 and 7 after ovulation, respectively. Estradiol secretion during the follicular phase was similar among groups. Onset of estrus (P<0.001) and the timing of maximum LH concentration (P<0.01) were earlier in group D3 than in D5 and D7 groups. During the subsequent cycle, the number and size of corpora lutea were higher (P<0.05) in ewes of the groups D3 (1.9+/-0.3 and 115.1+/-14.3mm(2)) and D5 (1.8+/-0.2 and 100.2+/-11.2mm(2)) than in group D7 (1.3+/-0.2 and 75.6+/-6.4mm(2)) group. In Experiment 2, 24 ewes were treated with two cloprostenol injections (7 days apart). Twelve ewes were exposed to "male effect" previous to an isolation period (ME group), whereas the remaining ewes were controls without male exposure (CTR group). Male effect induced earlier preovulatory LH surge (P<0.05) and ovulation (P<0.001) than CTR group. In Experiment 3, the estrus was synchronized in 68 ewes. Nineteen of them (group FGA) were treated using intravaginal sponges impregnated with fluorogestone acetate for 12 days and inseminated at 55h. Forty-nine females (group ME) were treated like ME group. Twenty-four (ME48 group) and 25 ewes (ME55 group) were inseminated at 48 and 55h after treatment, respectively. The fertility rate was numerically higher in ME48 than ME55 and FGA groups (62.5, 44.0 and 47.4%, respectively). In conclusions, the combined use of short-interval cloprostenol treatment and "male effect" may be an adequate alternative for synchronizing estrus and applying artificial insemination in hair sheep throughout the entire year. PMID- 19185341 TI - Influence of osmolality and ions on the activation and characteristics of zebrafish sperm motility. AB - Despite the prevalence of zebrafish as a model scientific organism, understanding sperm function in this species is essentially limited to observations that osmotic shock initiates motility. During natural spawning, sperm encounter a range of environmental salinities as well as freshwater mixed with egg-associated ovarian fluid (OF), thus sperm are likely to be exposed to saline prior to egg contact. Effects of saline on sperm function in this model species are unknown, but likely to be important. Using computer assisted sperm analysis, this study addressed the effects of osmolality of spawning media and ionic composition and pH on the proportion of sperm becoming motile at activation (motility), as well as sperm velocity and path. When activated with tap water, motility was maximal (80%) at 10s (earliest time measured), declining to 5% by 87 s postactivation. With activation at moderate osmolalities ( approximately 160-200 mmol/kg) initial motility was decreased relative to low osmolality, increased from 10 to 30s, and subsequently declined less rapidly (motility in 80 mM NaCl was 35%, 80%, and 60% at 10, 30 and 147 s, respectively). Thus, moderate osmolality increased duration, but introduced a temporal lag in motility onset. With moderate osmolalities, the rate of velocity decay was less than that with tap water activation. Sodium chloride and sucrose similarly impacted both motility and velocity. Replacement of NaCl with KCl, pH values ranging from 6.8 to 8.4, or the presence of gadolinium were without effect. Motility, but not velocity, was slightly supressed by Ca(2+). Therefore, whereas pH and concentrations of Ca(2+) or K(+) of OF are unlikely to impact fertility via sperm motility, the OF contribution to spawning media osmolality may have pronounced effects on motility and velocity of sperm, factors previously correlated with fertility in other species. PMID- 19185342 TI - Comparative efficacy and acceptability of 12 new-generation antidepressants: a multiple-treatments meta-analysis. AB - BACKGROUND: Conventional meta-analyses have shown inconsistent results for efficacy of second-generation antidepressants. We therefore did a multiple treatments meta-analysis, which accounts for both direct and indirect comparisons, to assess the effects of 12 new-generation antidepressants on major depression. METHODS: We systematically reviewed 117 randomised controlled trials (25 928 participants) from 1991 up to Nov 30, 2007, which compared any of the following antidepressants at therapeutic dose range for the acute treatment of unipolar major depression in adults: bupropion, citalopram, duloxetine, escitalopram, fluoxetine, fluvoxamine, milnacipran, mirtazapine, paroxetine, reboxetine, sertraline, and venlafaxine. The main outcomes were the proportion of patients who responded to or dropped out of the allocated treatment. Analysis was done on an intention-to-treat basis. FINDINGS: Mirtazapine, escitalopram, venlafaxine, and sertraline were significantly more efficacious than duloxetine (odds ratios [OR] 1.39, 1.33, 1.30 and 1.27, respectively), fluoxetine (1.37, 1.32, 1.28, and 1.25, respectively), fluvoxamine (1.41, 1.35, 1.30, and 1.27, respectively), paroxetine (1.35, 1.30, 1.27, and 1.22, respectively), and reboxetine (2.03, 1.95, 1.89, and 1.85, respectively). Reboxetine was significantly less efficacious than all the other antidepressants tested. Escitalopram and sertraline showed the best profile of acceptability, leading to significantly fewer discontinuations than did duloxetine, fluvoxamine, paroxetine, reboxetine, and venlafaxine. INTERPRETATION: Clinically important differences exist between commonly prescribed antidepressants for both efficacy and acceptability in favour of escitalopram and sertraline. Sertraline might be the best choice when starting treatment for moderate to severe major depression in adults because it has the most favourable balance between benefits, acceptability, and acquisition cost. PMID- 19185343 TI - Antidepressants are not all created equal. PMID- 19185344 TI - Hepcidin gene expression induced in the developmental stages of fish upon exposure to Benzo[a]pyrene (BaP). AB - Hepcidin is known to be expressed in fish with bacterial challenge and iron overload. Here we first report the hepcidin expression induced in the developmental stages from embryo to fry of red sea bream (Pagarus major) and in juvenile black porgy (Acanthopagrus schlegelii B.) upon continuous waterborne exposure to BaP. The gene expression of CYP1A1 and IgL (immunoglobulin light chain) were both measured. Expression of the Pagarus major hepcidin gene (PM hepc) was increased in post hatch fry at 24 h and 120 h exposure to BaP at concentrations of 0.1, 0.5 and 1.0 microg/l, respectively. The gene expression pattern was comparable to that of CYP1A1 but different from that of IgL. In addition, a high number of AS-hepc2 transcripts (Acanthopagrus schlegelii B. hepcidin gene) were detected in the liver upon exposure to 1.0 microg/l BaP. This study demonstrates that hepcidin gene expression is significantly induced in BaP exposed red sea bream and black porgy. PMID- 19185346 TI - Native and recombinant ASIC1a receptors conduct negligible Ca2+ entry. AB - Acid Sensing Ion Channels (ASICs) are a family of proton-gated cation channels that play a role in the sensation of noxious stimuli. Of these, ASIC1a is the only family member that is reported to be permeable to Ca(2+), although the absolute magnitude of the Ca(2+) current is unclear. Here, we used patch-clamp photometry to determine the contribution of Ca(2+) to total current through native and recombinant ASIC1a receptors. We found that acidification of the extracellular medium evoked amiloride and psalmotoxin 1-sensitive currents in isolated chick dorsal root ganglion neurons and human embryonic kidney cells, but did not alter fura-2 fluorescence when the bath concentration of Ca(2+) was close to that found in normal physiological conditions. Further, activation of recombinant ASIC1a receptors also failed to produce measurable changes in fluorescence despite of the fact that the total cation current through the over expressed receptor was ten-fold larger than that of the native channels. Finally, we imaged a field of intact DRG neurons loaded with the Ca(2+)-sensing dye Fluo 4, and found that acidification increased [Ca(2+)](i) in a small population of cells. Thus, although our whole-field imaging data agree with previous studies that activation of ASIC1a receptors can potentially cause elevations in intracellular free Ca(2+), our single cell data strongly challenges the view that Ca(2+) entry through the ASIC1a receptor itself contributes to this response. PMID- 19185345 TI - Biomimetic strategies based on viruses and bacteria for the development of immune evasive biomaterials. AB - The field of biomaterials has begun to focus upon materials strategies for modulating the immune response. While certain approaches appear promising, they are currently limited to isolated facets of inflammation process. It is well documented that both bacteria and viruses have highly developed methods for evading the immune system, providing inspiration for a more biomimetic approach to materials design. This review presents the immune evasive tactics employed by viruses and bacteria, and offers suggestions for future directions that apply these principles to design of immune evasive biomaterials. PMID- 19185347 TI - Mapping of IgE-binding epitopes on the major latex allergen Hev b 2 and the cross reacting 1,3beta-glucanase fruit allergens as a molecular basis for the latex fruit syndrome. AB - Nine distinct IgE-binding epitopes were identified along the entire amino acid sequence of the major latex allergen Hev b 2 (1,3beta-glucanase) using a set of synthetic 15-mer peptides frameshifted by 3 residues immobilized on cellulose membrane (Spot technique). Most of the amino acid residues building these IgE binding epitopic regions are nicely exposed on the surface and the epitopes usually correspond to charged regions on the molecular surface of the protein. A smaller number of 5 IgE-binding epitopic areas was identified on the banana 1,3beta-glucanase, which exhibits a very similar overall conformation and charge distribution. The latter epitopes might be responsible for the IgE-binding cross reactivity currently observed in the latex-fruit syndrome. Using rabbit polyclonal IgG anti-BanGluc as a probe instead of IgE from allergic patients the same epitopic regions were identified in both Hev b 2 and BanGluc. Additionally, surface-exposed regions with a very close conformation were predicted to occur on Ole e 9, the 1,3beta-glucanase allergen identified in olive pollen. PMID- 19185348 TI - Collagen regulates the activation of professional phagocytes of the teleost fish gilthead seabream. AB - The innate immune system mediates the initial inflammatory response that follows infection or injury. Although the innate immune response of fish to infection has been relatively well characterized during recent years at both cellular and molecular levels, no studies have examined the role of extracellular matrix (ECM) in the regulation of innate immunity and inflammation. We report here that collagen and gelatin in vitro were able to prime the respiratory burst of phagocytes from the bony fish gilthead seabream. In addition, collagen and gelatin induced a specific set of immune-related and ECM remodelling enzymes that substantially differed from that induced by pathogen-associated molecular patterns. Notably, both collagen and gelatin induced the expression of interleukin-1beta, chemokine (C-C motif) ligand 4 and matrix metalopeptidases (MMP) 9 and 13 in acidophilic granulocytes and macrophages but were unable to significantly increase the expression of other pro-inflammatory genes. Furthermore, it was found that the MMP2/MMP9 inhibitor V had a dose-dependent inhibitory effect on seabream phagocyte activation by either collagen or gelatin. In contrast, pre-treatment of collagen and gelatin by collagenase resulted in a higher stimulatory capacity compared to non-digested proteins. Collectively, these results indicate that collagen fragments produced by the action of different host proteases, and probably released by infectious agents, are sensed by fish phagocytes. Therefore, we propose that, besides to the well-established response to infection, the innate immune system of fish is able to respond to tissue injury. PMID- 19185349 TI - Standards of care for patients with invasive fungal infections within the United Kingdom: a national audit. AB - OBJECTIVE: The objective of this study was to audit the compliance and implementation of the British Society for Medical Mycology standards of care for patients with invasive infections in UK hospitals. METHODS: A multidisciplinary audit questionnaire regarding the processing of microbiology and histopathology specimens, radiology imaging and clinical management of patients with invasive fungal infections was distributed to UK hospitals. RESULTS: The study has shown that speciation of Candida and Aspergillus isolates from sterile sites was performed in 42-98% of hospitals. Microscopy of bronchoscopy specimens was not undertaken in 13 of 62 (21%) laboratories. Cryptococcal culture and antigen were undertaken routinely in abnormal CSF in 40-75% and 31-83% of at-risk patients but only in 12% of abnormal CSFs in patients without risk factors. Detailed fungal morphology was provided by <50% of histopathology departments. Most hospitals provided a timely HRCT or MRI on patients suspected to have an invasive fungal infection, but early treatment failed to occur in 15% of hospitals. In patients presenting with candidaemia, central venous catheters (CVC) were not changed routinely within 48h in 15%. CONCLUSION: Improvement in microbiology and histopathology specimen processing as well as rapid interventions such as initiation of anti-fungal therapy or CVC line removal could improve diagnostic rates and clinical outcomes of invasive fungal infections. PMID- 19185350 TI - Self-directed action affects planning in tool-use tasks with toddlers. AB - Toddlers grasp a tool more effectively when it is self-directed (e.g., spoon) than other-directed (e.g., hammer), possibly because the consequences of self directed actions are more obvious. When the negative consequences of an inefficient grip were made equally salient, the self-directed versus other directed differences remained. PMID- 19185351 TI - Long term developmental impact of social withdrawal in infants. AB - The aim of this study was to follow-up infants who were assessed in early infancy, at approximately 6 months of age, to determine the developmental impact of social withdrawal at approximately 30 months of age. Infants were administered the Bayley Scales of Infant Development-Third Edition, and the mothers complete the Behavior Assessment System for Children (BASC), Second Edition-Parent Rating Scale-Preschool form. Significant negative correlations were found between infant social withdrawal and Cognitive and Language scales of the Bayley Scales, and the Social and Communication scales of the BASC. Significant positive correlations were found between the infant social withdrawal and later higher scores on Atypicality and Attention scales. These results provide support for the use of the Alarm Distress Baby Scale as a measure of infant social withdrawal, and its association with later development and behavior. PMID- 19185352 TI - Procedural pain heart rate responses in massaged preterm infants. AB - Heart rate (HR) responses to the removal of a monitoring lead were assessed in 56 preterm infants who received moderate pressure, light pressure or no massage therapy. The infants who received moderate pressure massage therapy exhibited lower increases in HR suggesting an attenuated pain response. The heart rate of infants who received moderate pressure massage also returned to baseline faster than the heart rate of the other two groups, suggesting a faster recovery rate. PMID- 19185353 TI - Parenting of 7-month-old infants at familial risk for ADHD during infant's free play, with restrictions on interaction. AB - Patterns of interaction of 34 mothers and fathers with their 7-month-old boys at familial risk for ADHD and 25 comparison families were studied during infant play with blocks. The parents were instructed to refrain from intervening as much as possible. Infants in the risk group did not differ from those in the comparison group in frequency of needing help or involving parents in play. Nonetheless, they received adequate responsivity from their mothers less often than infants in the comparison group. Mothers in the risk group were also more likely not to respond to these needs at all. Mothers in the comparison group were more physically intrusive. No group difference was found for maternal rebuilding of the infant's play. No group differences were found for any of father's behaviors. However, fathers in both groups rebuilt their infant's play more frequently than mothers, infants looked at them more often, and a larger number of infants involved the father in their play. PMID- 19185354 TI - Screening for mood and anxiety disorders with the five-item, the three-item, and the two-item Mental Health Inventory. AB - The Mental Health Inventory (MHI)-5 is an attractive, brief screening questionnaire for depression and anxiety disorders. It has been suggested that the three questions on depression (MHI-d) may be as good as the full MHI-5 in assessing depressive disorders. We examined the validity of the MHI-d and the MHI a (the remaining two items on anxiety) in a large population-based sample of 7076 adults in the Netherlands. We also examined the validity of the MHI in assessing specific anxiety disorders. The presence of depressive and anxiety disorders in the past month was assessed with the Composite International Diagnostic Interview (CIDI), computerized version 1.1. ROC analyses indicated no significant difference between the MHI-5 (area under the curve of 0.93) and the MHI-d (area under the curve of 0.91) in detecting major depression and dysthymia. There was no difference either between the MHI-5 (area under the curve of 0.73) and the MHI a (area under the curve 0.73) in detecting anxiety disorders. Both the MHI-5 and the MHI-a also seem to be adequate as a screener for some anxiety disorders (generalized anxiety disorder; panic disorder; obsessive-compulsive disorder), but not others, especially phobias (agoraphobia; social phobia; simple phobia). PMID- 19185355 TI - Substance use disorders as risk factors for suicide in an Eastern and a Central European city (Tallinn and Frankfurt/Main). AB - Cultural and regional differences on the well-known elevated suicide risk in substance use disorders have not been clarified yet. Therefore, the suicide risk associated with substance use disorders in a society of transition and in a socially and economically stable society should be identified and compared. Data from two population-based matched case-control studies were used to analyse the association between alcohol and other substance consumption and the risk of suicide. Data in Frankfurt were obtained by a semi-structured interview including the Structured Clinical Interview for DSM-IV Axis I (SCID-I) in 163 suicides that occurred in 1999 and 2000, and data from Tallinn were collected according to DSM IV criteria on 156 deceased persons who committed suicide in 1999 by using the psychological autopsy method and in each city in matched population-based control persons by personal interview. In both cities, substance use disorders were significantly associated with suicide. Odds ratios for suicide were higher in Tallinn than in Frankfurt. The highest risk was observed in Tallinn among men with alcohol use disorders, aged 35 to 59 years. Although substance use and, in particular, alcohol use disorders were confirmed as risk factors for suicide in Tallinn and in Frankfurt, the much higher suicide risk associated with alcoholism in Tallinn than in Frankfurt indicates the importance of cultural, socio political, and regional impact of suicide risk in alcoholism. PMID- 19185356 TI - The origins of the International Journal of Pediatric Otorhinolaryngology. PMID- 19185357 TI - Tinnitus is prevalent in children with cochlear implants. AB - OBJECTIVES: To explore the prevalence and the perceived impact of tinnitus in children using cochlear implants. METHOD: Cross-sectional study of implanted children attending a cochlear implant family event organized annually by our academic tertiary pediatric care center. Children were interviewed together with their parents, using open-questioning and structured interview qualitative methodologies. The main outcome measures were the prevalence of tinnitus and any impact of these symptoms. RESULTS: 40 children (age range: 3-15, mean: 7 years) and their families were interviewed. These included unilateral implantees (n=21), and bilateral implantees (n=19) whose two procedures were simultaneous (n=6), within 6-12 months (n=3), or >2 years apart (n=10). Tinnitus was reported by 38% (n=15). Tinnitus occurred most commonly in the implanted ear, when the implants were not in use (e.g. in bed at night). The children were generally untroubled by the tinnitus, although two reported difficulty sleeping. Tinnitus was most frequent in children aged 6-8 years (8/17, 47%), and in bilateral implantees with an inter-procedure delay of at least 2 years (6/10, 60%). Tinnitus was least reported in those implanted bilaterally simultaneously (1/6, 17%), and in those 5 years old or younger (3/11, 27%). No obvious relationship was identified between the prevalence of tinnitus and the etiology of deafness, age of implantation, or time elapsed since implantation. CONCLUSIONS: To our knowledge this is the first study to report the widespread prevalence of tinnitus in implanted children. Further work, particularly examining the effect of inter-implant delay on tinnitus in bilateral implantees, may contribute to our understanding of the neuronal plasticity after implantation. PMID- 19185358 TI - In vitro and in vivo acute toxicity of fenpyroximate to flounder Paralichthys olivaceus and its gill cell line FG. AB - Fenpyroximate, an acaricide, is widely used in the prevention of acarids (mites) in plants. In this study, the in vitro and in vivo acute toxicity of fenpyroximate was examined using the marine flounder Paralichthys olivaceus and its gill cell line (FG). The 48h-IC(50) (95% confidence limits) values of fenpyroximate in the FG cells were 890 (790-990)nM, 950 (881-1019)nM and 1250 (1159-1341)nM, and 96h-IC(50) (95% confidence limits) were 480 (388-572)nM, 490 (454-526)nM and 510 (469-551)nM, for methyl thiazolyl tetrazolium (MTT) assay, neutral red (NR) uptake and cell protein assay, respectively. The 48h- and 96h LC(50) (95% confidence limits) values of fenpyroximate in living flounders were 28.84 (14.28-58.26)nM and 11.74 (6.06-22.8)nM, respectively. This indicated that fenpyroximate was highly toxic to both flounders and FG cells. Moreover, comparisons of the ratios of average 48h-IC(50) to 48h-LC(50) and average 96h IC(50) to 96h-LC(50) showed that the length of exposure time did not significantly affect the correlation between the FG cells and living flounders in the acute toxicity estimation of fenpyroximate provided the selected exposure time is the same. Thus, we suggest that FG cells could be a good bioassay system in rapid estimation of the corresponding LC(50) values of pollutants to living fish, instead of whole living fish. Histopathological examinations showed that liver and gill were the major target organs of fenpyroximate, especially the damage of gill tissues may account much for the high lethality of exposed flounders. Consistent with the histopathological observations, analysis of the activities of two key detoxification metabolism-related enzymes, glutathione S transferase (GST) and cytochrome P4501A1 (CYP1A1)-dependent ethoxyresorufin-O deethylase (EROD), in the liver and gill tissues of exposed flounders indicated that liver has much higher detoxification capacity than gills, and this contributes to the higher tolerance of liver to the toxicity of fenpyroximate in the exposed flounders. Fenpyroximate can initially induce a quick and significant increase of the activities of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) in all the exposed FG cells, and liver and gill tissues of exposed flounders. Upon continuation of the exposure the enzyme activities were inhibited, implying the occurrence of oxidative stress in the exposed fish cells and the possible interruption of the mitochondrial respiratory chain which involves redox reactions by fenpyroximate. PMID- 19185359 TI - Analysis of multimorbidity in individual elderly nursing home residents. Development of a multimorbidity matrix. AB - The chronic multimorbidity in individual elderly people is rarely documented in its entirety in present medical records, neither as cross-sectional overview nor as longitudinal time-course of various health problems. This obviously hampers an integrated clinical analysis. This work was aimed at evaluating the chronic multimorbidity in individual elderly patients and developing a method to map, quantify and grade the prevalence of the multimorbidity. An explorative study in 70 nursing home residents (55 women), mean age 85 was performed. Information on health problems was obtained through history, clinical examination and medical records. A 19-item multimorbidity matrix that maps, quantifies and grades the chronic morbidity in individual patients is presented. The 70 residents exhibited 275 different health problems; the top 3 items being neuropsychiatric, cardiovascular and gastrointestinal ones. The residents had a mean of 17 different chronic health problems and were prescribed a mean of 6.6 continuous medications per day. There was a significant correlation between the number of continuous drug prescriptions and both quantitative and graded multimorbidity scores. The presented multimorbidity matrix provides a useful taxonomic overview over the health situation in individual multimorbid elderly and constitutes the basis for ongoing work to develop and renew the electronic health record into an "interactive health analysis system". PMID- 19185360 TI - Tako-tsubo cardiomyopathy mimicking acute aortic dissection: a case report. AB - Tako-Tsubo cardiomyopathy is characterized by a transient and reversible left ventricular dysfunction possibly due to a catecholamine-mediated myocardial stunning. This form of cardiomyopathy is rarely associated with cardiogenic shock. Here we report on a patient with Tako-Tsubo cardiomyopathy induced low output failure in which blood flow disturbances in the thoracic aorta mimicked an acute type A aortic dissection. PMID- 19185361 TI - Takayasu arteritis presenting as type A intramural hematoma--an unusual presentation. AB - Takayasu arteritis which typically affects the aorta and its branches usually presents with obstructive lesions leading to ischemia. We report an unusual case of Takayasu arteritis presenting as an acute Type A intramural hematoma. PMID- 19185362 TI - Plasma indices of angiogenesis in rheumatoid disease: relationship to cardiovascular risk factors and cardiac function. AB - INTRODUCTION: Rheumatoid Disease (RD) is associated with increased rates of cardiovascular disease (CVD). Angiogenesis is central to RD, and well-recognized in CVD. We hypothesised that plasma levels of two indices associated with angiogenesis, vascular endothelial growth factor (VEGF) and angiogenin, would be higher among RD patients compared to healthy controls (HC), would relate to CVD risk factors, calculated 10-year coronary heart disease (CHD) and stroke risk scores. METHODS: 144 clinic patients with established RD and 63 HC were recruited in a cross-sectional study. RD patients were grouped according to the presence (RD-CVD, n=73 or absence (non-CVD RD; n=71) of CVD risk factors. Angiogenin and VEGF levels were quantified by ELISA. RESULTS: There were no significant differences for VEGF or angiogenin, between RD-CVD, non-CVD RD and HC groups (p=NS). Calculated risks for both CHD (p=0.017) and stroke (p=0.016) were higher when RD-CVD was compared to non-CVD RD and HC. Upon multivariate analysis, methotrexate use (p=0.006) and prior mycocardial infarction (MI) (p=0.034) were associated with higher angiogenin levels; body mass index (BMI) (p=0.034) and presence of RD (p=0.029) itself predicted lower levels. For RD patients, serum creatinine (p<0.001) and CRP levels, VEGF levels, and NSAID/COX2 inhibitor use (all p<0.05) were independently associated with CHD risk; plasma VEGF and serum creatinine levels were independently associated with stroke risk (p<0.05). CONCLUSIONS: Although levels of angiogenin were not significantly different between HC and RD patients, RD may have some influence on their variation. Methotrexate use and prior MI predicted higher angiogenin levels, whilst levels of VEGF were negatively associated with 10-year CHD and stroke risk. PMID- 19185363 TI - Anomalous origin and course of a dual left anterior descending coronary artery. AB - Dual left anterior descending artery (LAD) is a rare coronary artery anomaly. Dual but normally originated LAD has been usually reported to have no clinical significance. In this case report, we present a case of a middle age year old male with atypical anginal symptoms in whom coronary arteriography showed dual LAD with anomalous origin of one branch from the right coronary artery. PMID- 19185364 TI - Erythropoietin levels in cardiac resynchronization therapy responders. AB - Cardiac resynchronization therapy (CRT) is an effective treatment strategy in patients with severe left ventricular dysfunction and those with evidence of electro-mechanical dyssynchrony. A prospective observational study was conducted in patients with severe symptomatic heart failure undergoing CRT implantation. Thirty patients were classified as either responders (9 patients), or non responders (21 patients). The association between functional status, quality of life scores and CRT responder status with erythropoietin (EPO) levels was determined. The mean EPO levels among responders were 28.8+/-3.6 and 11.4+/-1.7 mIU/ml among non-responders. Elevated EPO levels in the absence of anemia and renal insufficiency effectively correlate with functional impairment and severe symptoms, which impair quality of life in patients undergoing CRT. PMID- 19185365 TI - Atrial fibrillation associated with subclinical hyperthyroidism. AB - Subclinical hyperthyroidism is an increasingly recognized entity that is defined as a normal serum free thyroxine and free triiodothyronine levels with a thyroid stimulating hormone level suppressed below the normal range and usually undetectable. It has been reported that subclinical hyperthyroidism is not associated with coronary heart disease or mortality from cardiovascular causes but it is sufficient to induce arrhythmias including atrial fibrillation and atrial flutter. It has also been reported that increased factor X activity in patients with subclinical hyperthyroidism represents a potential hypercoagulable state. We present a case of atrial fibrillation associated with subclinical hyperthyroidism, in a 78-year-old Italian woman. Also this case focuses attention on the importance of a correct evaluation of subclinical hyperthyroidism. PMID- 19185366 TI - A case of very late thrombosis of bare metal stent successfully treated with excimer laser coronary angioplasty. AB - Late thrombosis occurring after bare metal stent (BMS) implantation is a rare complication. However, it is often associated with poor outcome. Mechanisms underlying BMS thrombosis may differ from those underlying drug eluting stent thrombosis. In particular, severe instent restenosis may trigger thrombus formation. This case report describes a very late thrombosis of BMS occurring on an in-stent restenosis. After failure of manual thrombectomy, the case was successfully faced by Excimer Laser Coronary Angioplasty (ELCA). Cases like this may be successfully treated by devices like ELCA combining debulking and thrombus removal capabilities. PMID- 19185367 TI - Justice where justice is due: A posthumous Nobel Prize to Carlos Chagas (1879 1934), the discoverer of American Trypanosomiasis (Chagas' disease). AB - Working in the Brazilian backland, Chagas described a new disease. He discovered the etiologic agent, the vector, the reservoir, the acute stage, the several clinical aspects of the chronic stage (particularly the heart disease), role of autoimmunity in its pathogenesis, and anticipated the social impact of the disease. Chagas was nominated to Nobel Prize twice: in 1913, and in 1921. In 1913, Richet won the prize because his work on anaphylaxis. In 1921, no one received the Nobel Prize. It is believed that detraction of Chagas' work at the National Academy of Medicine, made by jealousy, mediocrity, and political rivalries can be maculated the image of the scientist. Furthermore, misperception of Chagas' work may also have led the Nobel Committee not to award him. One hundred years after the discovery, we can appreciate the greatness of the discovery of Carlos Chagas, never seem in the realm of biological research. Time to make justice, therefore, has finally come. PMID- 19185368 TI - A possible diagnosis of isolated right ventricular hypertrabeculation /noncompaction in an elderly man by three-dimensional echocardiography. AB - A 88 year-old man admitted with iterative severe chest distress and breathlessness associated with activity without chest complaint for 5 years. He underwent an initial 2DE that did not confirm HVM/NVM and 3DE that strengthened the diagnosis of HVM/NVM. In contrast to 2DE, 3DE provides for pyramid-shaped datasets that encompass the entire right ventricular. Specifically, right ventricular can be sectioned in user selected planes and an unlimited number and angles of such planes can be used. The present case should be regarded as isolated right ventricular HVM/NVM because there were no any factors that could explain the arrest of the development of the myocardial structure. PMID- 19185369 TI - 2-Dimensional strain echocardiography and early detection of myocardial ischemia. AB - Echocardiographic strain with strain rate imaging is a new technology enabling more reliable and comprehensive assessment of myocardial function. The spectrum of potential clinical applications is very wide due to its ability to differentiate between active and passive movement of myocardial segments, to quantify intraventricular dyssynchrony and to evaluate components of myocardial function, such as longitudinal myocardial shortening, that are not visually assessable. In-vivo and in-vitro validation of 2D-strain imaging technique have been undertaken and reached a point where it is considered ready for more widespread investigations into clinical utility, e.g. regarding myocarditis, arrhythmogenic right ventricular dysplasia/cardiomyopathy and regional ischemia. Moreover, longitudinal LV strain is closely related to log plasma brain-type natriuretic peptide levels in patients with congestive heart failure, both in patients with systolic and diastolic heart failure. We present a case of detection of coronary artery disease in a 55-year-old Italian man. This case focuses attention on the higher sensibility of the 2-Dimensional Strain echocardiography the diagnosis of myocardial ischemia in patients with coronary artery disease. PMID- 19185370 TI - Complex arrhythmias in Prinzmetal's angina reproduced by cold pressor test. AB - We describe the case of a young female patient with syncope due to complex arrhythmias associated with Prinzmetal's angina. The clinical situation was reproduced by the cold pressor test. PMID- 19185371 TI - Delayed enhancement cardiovascular magnetic resonance as a novel technique to predict cardiac events in dilated cardiomyopathy patients. AB - PURPOSE: Delayed enhancement cardiovascular magnetic resonance (DE-CMR) can detect cardiac scarring and has the potential to visualize the progression of myocardial remodeling. We determined whether DE-CMR can predict cardiac events in dilated cardiomyopathy patients. MATERIALS AND METHODS: Transthoracic echocardiography, coronary arteriography, and DE-CMR studies were performed in 60 consecutive dilated cardiomyopathy (DCM) patients. Percent delayed enhancement (%DE) was determined as the ratio of the area showing delayed enhancement to the total myocardial area in three short-axis views. Patients were classified as advanced group (Group A) when %DE was 10% or higher, and as non-advanced group (Group NA) when %DE was less than 10%. The incidence of cardiac events and the clinical history were compared between Group A and Group NA. RESULTS: There were 11 patients in Group A and 49 patients in Group NA. The incidence of cardiac events was significantly higher in Group A (36%; 4/11 patients) than in Group NA (2.0%; 1/49 patients) (log rank, p=0.0001). CONCLUSION: DE-CMR is a useful tool to predict cardiac events in DCM patients. PMID- 19185372 TI - Impact of history of heart failure on diagnostic and prognostic value of BNP: results from the B-type Natriuretic Peptide for Acute Shortness of Breath Evaluation (BASEL) study. AB - OBJECTIVES: This study aimed to examine the influence of history of heart failure (HF) on circulating levels, diagnostic accuracy and prognostic value of B-type natriuretic peptide (BNP) in patients presenting with all cause dyspnea at the emergency department. BACKGROUND: BNP has been shown to be very helpful in diagnosis and prognosis of HF. Due to chronically elevated cardiac filling pressures, patients with a history of HF might have higher BNP levels and therefore diagnostic and prognostic properties of BNP may be affected. METHODS: We analyzed circulating levels, diagnostic accuracy and prognostic value of BNP in 388 patients without a previous history of HF and compared these to data to 64 patients with a history of HF included in the B-type Natriuretic Peptide for Acute Shortness of Breath Evaluation (BASEL) Study. RESULTS: Baseline BNP levels were higher in patients with a history of HF (median 814 pg/ml [353-1300 pg/ml] vs. 216 pg/ml [45-801 pg/ml], p<0.001). Diagnostic accuracy of BNP to identify HF was comparable in patients with (AUC=0.804; 95% CI 0.628-0.980) and in patients without history of HF (AUC=0.883; 95% CI 0.848-0.919, p=0.389). Prognostic ability of BNP to predict one-year mortality was lower in overall patients with history of HF (AUC=0.458; 95%CI 0.294-0.622) compared to patients without history of HF (AUC=0.710; 95% CI 0.653-0.768, p<0.05). CONCLUSIONS: In patients with history of HF, BNP levels retain diagnostic accuracy. Ability to predict one-year mortality was decreased in unselected patients, but not in patients with acute HF induced dyspnea. PMID- 19185375 TI - A golden opportunity, squandered. PMID- 19185373 TI - Isolated cleft of the posterior mitral valve leaflet in a patient with Marfan syndrome. AB - Isolated cleft of the mitral valve is an uncommon congenital cause of mitral regurgitation. Most of the clefts involve the anterior leaflet and isolated clefts of the posterior leaflet have been anecdotally identified. We report a case of Marfan syndrome in whom an echocardiographic examination revealed a cleft in the posterior mitral leaflet. Marfan syndrome frequently involves the mitral valve causing mitral valve prolapse. Occurrence of a cleft may be an extension of its mitral valve involvement. PMID- 19185376 TI - Efficiency of slightly acidic electrolyzed water for inactivation of Salmonella enteritidis and its contaminated shell eggs. AB - The efficiency of slightly acidic electrolyzed water (SAEW) at different temperatures (4, 20 and 45 degrees C) for inactivation of Salmonella enteritidis and it on the surface of shell eggs was evaluated. The bactericidal activity of SAEW, sodium hypochlorite solution (NaClO) and acidic electrolyzed water (AEW) to inactivate S. enteritidis was also compared. SAEW with a pH value of 6.0-6.5 used was generated by the electrolysis of a dilute hydrochloric acid (2.4 mM) in a chamber without a membrane. Although the pH value of SAEW was greatly higher than that of AEW (pH2.6-2.7), SAEW had a comparative powerful bactericidal activity at the same available chlorine concentrations. The efficiency of SAEW for inactivation of pure S. enteritidis cultures increased with increasing the available chlorine concentration and treatment time at the three different temperatures. The S. enteritidis counts decreased to less than 1.0 log(10) CFU/ml at available chlorine of 2 mg/l and 100% inactivation (reduction of 8.2 log(10) CFU/ml) was resulted in using SAEW with available chlorine more than 4 mg/l at 4, 20 and 45 degrees C after 2 min treatment, whereas no reduction was observed in the control samples. Moreover, SAEW was also effective for inactivating the S. enteritidis inoculated on the surface of shell eggs. A reduction of 6.5 log(10) CFU/g of S. enteritidis on shell eggs was achieved by SAEW containing 15 mg/l available chlorine for 3 min, but only a reduction of 0.9-1.2 log(10) CFU/g for the control samples. No survival of S. enteritidis was recovered in waste wash SAEW after treatment. The findings of this study indicate that SAEW may be a promising disinfectant agent for the shell egg washing processing without environmental pollution. PMID- 19185377 TI - Microfluidic technology applied to cell-wall protein analysis of olive related lactic acid bacteria. AB - A fully automated electrophoretic method for separating cell-wall proteins was applied to 102 isolates from olive phylloplane and brine previously phenotypically identified as Lactobacillus plantarum (10), Leuconostoc mesenteroides subsp. mesenteroides (60), Leuconostoc spp. (12) and Enterococcus faecium (19). The high sensitivity of this technique allowed to discriminate isolates and to distinguish closely related strains within the same species. L. plantarum was characterized by proteins at 70.4 and 163.9 kDa, while 3 proteins (20.4, 52.9, 68. kDa) were typical for E. faecium. No characteristic proteins were found for Leuconostoc isolates except for that at 31.8 kDa mainly related to the ecological origin since only isolates from brine showed this protein. However, Leuc. mesenteroides subsp. mesenteroides and Leuconostoc spp. were differentiated by additional proteins. Sequence analysis of the 16S rRNA gene of selected strains was in agreement with the phenotypic identification and allowed to identify all Leuconostoc spp. isolates as Leuc. mesenteroides. Cell-wall protein fingerprinting performed by microfluidic technology completed the phenotypic characterization of olive-related lactic acid bacteria allowing their strain typing. PMID- 19185378 TI - Potato carrot agar with manganese as an isolation medium for Alternaria, Epicoccum and Phoma. AB - A semi-selective medium for isolation of Alternaria spp., Epicoccum sp. and Phoma spp. from soil and plant samples was developed. The basal medium was a modified potato carrot agar (PCA), containing 10 g/L of potato and carrot. It is known that the target genera sporulate well on standard PCA when grown at 25 degrees C with an alternating light/dark cycle consisting of 8 h of cool-white daylight followed by 16 h darkness. Addition of 1.5% MnCl(2) 4 H(2)O (w/v) inhibited most other fungi than Alternaria, Epicoccum and Phoma species when tested on pure cultures. The mycobiota of two soil samples and eight grain samples were examined using PCA-Mn and three commonly used isolation media, DRYES, DG18 and V8. On the three conventional media growth of several genera was observed with the predominant being Aspergillus, Eurotium, Fusarium, Mucor, Penicillium and Rhizopus. Of these only F. oxysporum and F. verticillioides were able to grow on PCA-Mn. Alternaria infectoria and Epicoccum nigrum were present in three cereal grain samples, but emerged to a far lower degree on the three conventional media compared to PCA-Mn. Three black spored fungi, identified as Phoma eupyrena, Paraconiothyrium minitan and one unknown species, were isolated from the two soil samples when incubated on PCA-Mn but were absent on the three conventional media. PMID- 19185379 TI - Time and temperature dependent microbiological and mycotoxin (ochratoxin-A) levels in boza. AB - This study describes the examination of microbiological tests and the determination of OTA in boza temperature and time dependently. Prior to the analysis, physicochemical properties of the boza samples such as moisture, total acidity as lactic acid, pH, protein amount and viscosity were investigated. The incidence of total aerobic bacteria (TAB), lactic acid bacteria (LAB), coliforms, E.coli, Salmonella, S. aureus, B. cereus, yeast and moulds were examined. E.coli, Salmonella, S. aureus and B. cereus were not found in all boza samples. Initially, Aspergillus fumigatus; Acremonium sp.; Geotrichum candidum and Geotrichum capitatum were identified in the samples. Certain extraction techniques such as direct injection, liquid-liquid and solid phase (SP) were tried for the OTA analysis. The most available way was found to be direct injection among them and the recovery was 70.56%+/-9.80 (13.89 RSD). OTA amounts were determined in all boza samples utilizing an isocratic HPLC analysis with an ODS column. OTA was detected in only one sample as 3.58 microg/kg and this amount is above the limits of European Commission Regulations. Time and temperature dependent changes were investigated and insignificant variation was observed. PMID- 19185380 TI - Endothelial dysfunction and cardiovascular risk factors. AB - The endothelium plays an integral role in the regulation of vascular tone, platelet activity, leukocyte adhesion, and thrombosis and is intimately involved in the development of atherosclerosis. Endothelial dysfunction has been observed in patients with established coronary artery disease or coronary risk factors, both in the coronary and peripheral vasculature. Therapeutic interventions with lipid-lowering drugs, ACE inhibitors, physical activity, and antioxidant agents have been shown to improve endothelial function in coronary and peripheral vessels. This systemic manifestation and improvement of endothelial function suggests that a common mechanism may contribute to endothelial dysfunction in the coronary and peripheral circulation. TARGET AUDIENCE: Internist, Cardiologists, Family physicians. LEARNING OBJECTIVES: After completion of this article, the reader should be able to define the participation of cardiovascular risk factors in the various complications associated with endothelial dysfunction. PMID- 19185381 TI - The Leu262Val polymorphism of presenilin associated rhomboid like protein (PARL) is associated with earlier onset of type 2 diabetes and increased urinary microalbumin creatinine ratio in an Irish case-control population. AB - AIMS: Environmental and genetic factors contribute to the evolution of type 2 diabetes (T2DM). Presenilin associated rhomboid like protein (PARL) is a mitochondrial protein that has been implicated in T2DM in both the rodent Psammomys obesus and in humans. The SNP variant (Leu262Val) in PARL has been shown to be associated with hyperinsulinaemia in an age-dependent manner in a US non-diabetic, cohort. However, this finding has not been replicated in UK cohorts. We studied Leu262Val associations in an Irish Caucasian T2DM case control population. METHODS: An RFLP-PCR assay using BstN I was used to assess Leu262Val genotype in a total of 613 subjects, 421 with T2DM and 192 controls. RESULTS: In the control group genotype frequencies were as follows 27.37% (GG), 51.58% (CG) and 21.05% (CC), while in the group with T2DM 30.64% (GG), 47.74% (CG) and 21.62% (CC). We observed no association between Leu262Val variant and T2DM nor was there an association with plasma insulin concentrations or BMI. There was no interaction between age and fasting plasma insulin concentration. However, in the group with T2DM the C allele was associated with higher urinary albumin to creatinine ratio while the GG genotype was associated with an earlier age of onset of T2DM. CONCLUSION: The Leu262Val polymorphism of PARL is not associated with markers of insulin resistance. However, in subjects with T2DM, genetic variation at this locus may indicate earlier onset of T2DM and increased susceptibility to nephropathy and cardiovascular complications. PMID- 19185382 TI - Genetics and epigenetics: stability and plasticity during cellular differentiation. AB - Stem cells and multipotent progenitor cells face the challenge of balancing the stability and plasticity of their developmental states. Their self-renewal requires the maintenance of a defined gene-expression program, which must be stably adjusted towards a new fate upon differentiation. Recent data imply that epigenetic mechanisms can confer robustness to steady state gene expression but can also direct the terminal fate of lineage-restricted multipotent progenitor cells. Here, we review the latest models for how changes in chromatin and DNA methylation are regulated during cellular differentiation. We further propose that targets of epigenetic repression share common features in the sequences of their regulatory regions, thereby suggesting a co-evolution of epigenetic pathways and classes of cis-acting elements. PMID- 19185383 TI - Oxygen uptake kinetics during cardiopulmonary exercise testing and postoperative complications in patients with lung cancer. AB - It is accepted that cardiopulmonary exercise testing is one of the most valuable parameters, especially peak oxygen uptake (VO(2)), for the evaluation of risk assessment in lung cancer surgery. It therefore represents an attractive way of identifying a patient at high risk for postoperative complications. However, many patients do not achieve the maximal or predictive level during an incremental exercise testing. The purpose of the current investigation was to study the value of the oxygen uptake efficiency slope (OUES), which shows exercise capacity during submaximal testing, in predicting postoperative mortality in patients with bronchogenic carcinoma scheduled for lung resection. Forty-nine patients with bronchogenic carcinoma participated in studies with exercise tests as a preoperative evaluation. The peak VO(2) was calculated for each subject by averaging values obtained during the final 10s of exercise. The following equation was used to determine OUES: VO(2)/log(10)VE. Peak VO(2) without postoperative complication was 22.8+/-3.3 ml/(kg min), however, peak VO(2) in patients with present complications was 19.1+/-4.2 ml/(kg min) (p=0.001). In addition, although the mean OUES in patients with present complications was 11.1+/-1.2, the mean OUES in the absent group was 13.3+/-2.1 (p<0.001). Although peak VO(2) is useful in evaluating selected patients with bronchogenic carcinoma, OUES is also a beneficial parameter and should be calculated and recorded with peak VO(2), a better predictor of poor surgical outcome than absolute values, and should be integrated into preoperative decision making. PMID- 19185384 TI - Necessity of centralisation of EBUS. AB - In the study of Bauwens et al. endobronchial ultrasound (EBUS) was provocated as first step procedure in the staging of PET mediastinal hot spots in lung cancer patients. In case of negative findings a surgical procedure should be undertaken. We certainly agree that in case of a negative finding a surgical procedure should be performed, however, we disagree that the first step procedure should be EBUS. In our opinion the first step procedure in a standard clinical practice should be a standard transbronchial needle aspiration (TBNA). PMID- 19185385 TI - Gefitinib as targeted therapy for mucoepidermoid carcinoma of the lung: possible significance of CRTC1-MAML2 oncogene. AB - Recent reports of a clinical response to gefitinib in pulmonary mucoepidermoid carcinoma (MEC) in the absence of sensitizing EGFR mutations suggest that tyrosine kinase inhibitors (TKIs) may be effective in this tumor type. Although not documented in these reports, MEC of the lung may harbor a t(11;19) translocation with an associated novel fusion oncogene (CRTC1-MAML2). Furthermore, MECs arising in the salivary glands carry this mutation in a high proportion of cases. In vitro data has shown that MEC cell-lines with t(11;19) are sensitive to gefitinib and that this may be mediated by the action of CRTC1 MAML2 in up-regulating the EGFR ligand, amphiregulin. Data also shows that gefitinib demonstrates amphiregulin-dependant activity in NSCLC cell-lines. As such, it may be speculated that MEC from lung and salivary glands expressing CRTC1-MAML2 present a valid target for treatment with gefitinib, even in the absence of sensitizing EGFR mutations. Clinical studies are required to test this hypothesis. PMID- 19185386 TI - Generalized linear mixed models: a practical guide for ecology and evolution. AB - How should ecologists and evolutionary biologists analyze nonnormal data that involve random effects? Nonnormal data such as counts or proportions often defy classical statistical procedures. Generalized linear mixed models (GLMMs) provide a more flexible approach for analyzing nonnormal data when random effects are present. The explosion of research on GLMMs in the last decade has generated considerable uncertainty for practitioners in ecology and evolution. Despite the availability of accurate techniques for estimating GLMM parameters in simple cases, complex GLMMs are challenging to fit and statistical inference such as hypothesis testing remains difficult. We review the use (and misuse) of GLMMs in ecology and evolution, discuss estimation and inference and summarize 'best practice' data analysis procedures for scientists facing this challenge. PMID- 19185387 TI - Salicylic acid improves acclimation to salt stress by stimulating abscisic aldehyde oxidase activity and abscisic acid accumulation, and increases Na+ content in leaves without toxicity symptoms in Solanum lycopersicum L. AB - Pre-treatment with 10(-4)M salicylic acid (SA) in hydroponic culture medium provided protection against salinity stress in tomato plants (Solanum lycopersicum L. cv. Rio Fuego). The effect of 10(-7) or 10(-4)M SA on the water status of plants was examined in relation to the biosynthesis and accumulation of abscisic acid (ABA) in order to reveal the role of SA in the subsequent response to salt stress. Both pre-treatments inhibited the K+(86Rb+) uptake of plants, reduced the K+ content of leaves, and caused a decrease in leaf water potential (psi(w)). Due to the changes in the cellular water status, SA triggered the accumulation of ABA. Since the decrease in psi(w) proved to be transient, the effect of SA on ABA synthesis may also develop via other mechanisms. In spite of osmotic adaptation, the application of 10(-4)M, but not 10(-7)M SA, led to prolonged ABA accumulation and to enhanced activity of aldehyde oxidase (AO1, EC.1.2.3.1.), an enzyme responsible for the conversion of ABA-aldehyde to ABA, both in root and leaf tissues. AO2-AO4 isoforms from the root extracts also exhibited increased activities. The fact that the activities of AO are significantly enhanced both in the leaves and roots of plants exposed to 10(-4)M SA, may indicate a positive feedback regulation of ABA synthesis by ABA in this system. Moreover, during a 100mM NaCl treatment, higher levels of free putrescine or spermine were found in these leaves or roots, respectively, than in the salt stressed controls, suggesting that polyamines may be implicated in the protection response of the cells. As a result, Na+ could be transported to the leaf mesophyll cells without known symptoms of salt toxicity. PMID- 19185388 TI - Nickel-induced changes in nitrogen metabolism in wheat shoots. AB - The influences of 50 and 100muM Ni on growth, tissue Ni accumulation, concentrations of nitrate, ammonium, glutamate, and proline as well as the activities of nitrate reductase (NR), nitrite reductase (NiR), glutamine synthetase (GS), glutamate synthase (GOGAT), glutamate dehydrogenase (GDH), alanine aminotransferase (AlaAT), and aspartate aminotransferase (AspAT) were examined in the shoots of wheat seedlings cv. Zyta. Exposure of the seedlings to Ni resulted in a rapid accumulation of this metal in the shoots, which was accompanied by significant reduction in fresh weight of these organs. Tissue nitrate content decreased in response to Ni stress, while ammonium concentration increased substantially. Glutamate concentration was slightly lowered up to the 4th day of the metal exposure. In contrast, proline content increased significantly, starting from the first day after Ni treatment. NR activity showed a decline of up to 40% below the control level after Ni application; however, its activation state remained unaltered. Heavy metal treatment also resulted in a marked decrease in NiR activity, which after 7d of exposure to 100muM Ni was almost 80% lower than in the control. GS activity in wheat shoots was not influenced by Ni application. Contrary to Fd-GOGAT exhibiting reduced activity in the shoots of Ni-treated wheat seedlings, NADH-GOGAT activity was considerably enhanced, exceeding the control value even by 165%. After 7d of exposure to Ni, both NADH-GDH and NAD-GDH activities in wheat shoots were markedly induced; however, NAD-GDH activity showed a significant decrease at the early stage of the experiment. Both AlaAT and AspAT glutamate-producing activities were considerably stimulated by Ni treatment. Our results suggest that induction of NADH-GOGAT, NADH-GDH, AlaAT, and AspAT activities may compensate for the reduced Fd-GOGAT activity and serve as an alternative means of glutamate synthesis in wheat shoots under Ni stress. PMID- 19185389 TI - Functional analysis of the buckwheat metallothionein promoter: tissue specificity pattern and up-regulation under complex stress stimuli. AB - To shed light on expression regulation of the metallothionein gene from buckwheat (FeMT3), functional promoter analysis was performed with a complete 5' regulatory region and two deletion variants, employing stably transformed tobacco plants. Histochemical GUS assay of transgenic tobacco lines showed the strongest signals in vascular elements of leaves and in pollen grains, while somewhat weaker staining was observed in the roots of mature plants. This tissue specificity pattern implies a possible function of buckwheat MT3 in those tissues. Quantitative GUS assay showed strong up-regulation of all three promoter constructs (proportional to the length of the regulatory region) in leaves submerged in liquid MS medium containing sucrose, after a prolonged time period. This represented a complex stress situation composed of several synergistically related stress stimuli. These findings suggest complex transcriptional regulation of FeMT3, requiring interactions among a number of different factors. PMID- 19185390 TI - Serious bacterial infections in infants who have experienced an apparent life threatening event. AB - STUDY OBJECTIVE: We determine the incidence of serious bacterial infection in infants presenting to the emergency department (ED) with an apparent life threatening event. METHODS: This was a prospective cohort study of infants younger than 12 months who were treated at an urban children's hospital ED during 1 year and who met standard criteria for an apparent life-threatening event. Sociodemographic data, presenting features, diagnostic testing, hospital course, and final diagnosis were collected. Follow-up information was obtained 4 weeks after the visit. RESULTS: Of the 198 infants enrolled, 44 (22.2%) had evaluation for serious bacterial infection: urine culture 36 (18%), blood culture 38 (19%) and cerebrospinal fluid culture 18 (9%). No infant (0/198; 95% confidence interval 0% to 1.8%) was found to have bacteremia, urinary tract infection, or bacterial meningitis, which was confirmed at the 4-week interview (99% ascertainment). Two infants were found to have enteroviral meningitis. CONCLUSION: The incidence of serious bacterial infection in this cohort was extremely low. Infants presenting with an apparent life-threatening event need not be subjected to a full serious bacterial infection evaluation routinely. PMID- 19185391 TI - Evidence-based Emergency Medicine/Rational Clinical Examination Abstract. Does the clinical examination predict lower extremity peripheral arterial disease? PMID- 19185392 TI - Interrater reliability and accuracy of clinicians and trained research assistants performing prospective data collection in emergency department patients with potential acute coronary syndrome. AB - STUDY OBJECTIVE: Clinical research requires high-quality data collection. Data collected at the emergency department evaluation is generally considered more precise than data collected through chart abstraction but is cumbersome and time consuming. We test whether trained research assistants without a medical background can obtain clinical research data as accurately as physicians. We hypothesize that they would be at least as accurate because they would not be distracted by clinical requirements. METHODS: We conducted a prospective comparative study of 33 trained research assistants and 39 physicians (35 residents) to assess interrater reliability with respect to guideline-recommended clinical research data. Immediately after the research assistant and clinician evaluation, the data were compared by a tiebreaker third person who forced the patient to choose one of the 2 answers as the correct one when responses were discordant. Crude percentage agreement and interrater reliability were assessed (kappa statistic). RESULTS: One hundred forty-three patients were recruited (mean age 50.7 years; 47% female patients). Overall, the median agreement was 81% (interquartile range [IQR] 73% to 92%) and interrater reliability was fair (kappa value 0.36 [IQR 0.26 to 0.52]) but varied across categories of data: cardiac risk factors (median 86% [IQR 81% to 93%]; median 0.69 [IQR 0.62 to 0.83]), other cardiac history (median 93% [IQR 79% to 95%]; median 0.56 [IQR 0.29 to 0.77]), pain location (median 92% [IR 86% to 94%]; median 0.37 [IQR 0.25 to 0.29]), radiation (median 86% [IQR 85% to 87%]; median 0.37 [IQR 0.26 to 0.42]), quality (median 85% [IQR 75% to 94%]; median 0.29 [IQR 0.23 to 0.40]), and associated symptoms (median 74% [IQR 65% to 78%]; median 0.28 [IQR 0.20 to 0.40]). When discordant information was obtained, the research assistant was more often correct (median 64% [IQR 53% to 72%]). CONCLUSION: The relatively fair interrater reliability observed in our study is consistent with previous studies evaluating interrater reliability for cardiovascular disease in the inpatient setting. With respect to research data, we found that prospective ascertainment of clinical data is more often correct when done by research assistants compared with clinicians simultaneously evaluating patients. PMID- 19185393 TI - Hospice and palliative medicine: new subspecialty, new opportunities. AB - Palliative care is the physical, psychological, social, and spiritual care provided to patients from diagnosis to death or resolution of a life-threatening illness. Hospice care is a comprehensive program of care that is appropriate when patients with chronic, progressive, and eventually fatal illness are determined to have a prognosis of 6 months or fewer. Hospice and palliative medicine has now been recognized by the American Board of Medical Subspecialties as a field with a unique body of knowledge and practice. With 9 other specialty boards, the American Board of Emergency Medicine has cosponsored hospice and palliative medicine as an official subspecialty. As a result, board-certified emergency physicians may now pursue certification in hospice and palliative medicine through either fellowship training or, for a limited time, completing practice track requirements, followed by a written examination in the subspecialty. As the practice of palliative medicine grows in hospitals, emergency physicians can develop a core of generalist palliative medicine skills for use with adults and children. These would include assessing and communicating prognoses, managing the relief of pain and other distressing symptoms, helping articulate goals of patient care, understanding ethical and legal requirements; and ensuring the provision of culturally appropriate spiritual care in the last hours of living. Front-line emergency physicians possessing these basic palliative medicine skills will be able to work collaboratively with subspecialty physicians who are dually certified in emergency medicine and hospice and palliative medicine. Together, generalist and specialist emergency physicians can advance research, education, and policy in this new field to reach the common goals of high-quality, efficient, evidence-based palliative care in the emergency department. PMID- 19185394 TI - Ethanol-based cleanser versus isopropyl alcohol to decontaminate stethoscopes. AB - Approximately 1 in 20 hospital admissions is complicated by a health care associated infection. Stethoscopes may play a role in spreading nosocomial infections. The objective of this study was to determine the effectiveness of an ethanol-based cleanser (EBC) compared with isopropyl alcohol pads in reducing bacterial contamination of stethoscope diaphragms. Stethoscopes were cultured from medical professionals on 4 medical floors before and after cleaning with either EBC or isopropyl alcohol pads. The numbers of colony-forming units (cfu) grown were compared between the 2 cleaners and to baseline values. A total of 99 stethoscopes were cultured (49 EBC; 50 isopropyl alcohol), and all were positive for growth. After cleaning, 28.28% of the stethoscopes were growth-free (12 EBC; 16 isopropyl alcohol). Cleaning with EBC and isopropyl alcohol pads significantly reduced the cfu counts (by 92.8% and 92.5%, respectively), but neither was found to be statistically superior (F = 1.22; P = .2721). Cleaning a stethoscope diaphragm using either EBC or isopropyl alcohol led to a significant reduction in bacterial growth in culture. As an extension of the hand, a stethoscope should be cleaned with the same frequency as the hands. The simultaneous cleaning of hands and stethoscope may further increase compliance with current standards. PMID- 19185395 TI - VEGF protects motor neurons against excitotoxicity by upregulation of GluR2. AB - Influx of Ca(2+) ions through the alpha-amino-3-hydroxy-5-methylisoxazole propionic acid (AMPA) receptors is toxic to neurons and contributes to motor neuron degeneration observed in amyotrophic lateral sclerosis (ALS). The Ca(2+) permeability of the AMPA receptor depends on its subunit composition. If the GluR2 subunit is present in the receptor complex, the AMPA receptor is impermeable to Ca(2+). In this study, we identified vascular endothelial growth factor-A (VEGF) as a GluR2 inducing molecule. Cultured motor neurons pretreated with VEGF displayed higher GluR2 levels. This resulted in AMPA receptor currents with a low relative Ca(2+) permeability and in motor neurons that were less vulnerable to AMPA receptor-mediated excitotoxicity. This effect of VEGF was mediated through the VEGFR2 present on the motor neurons and was due to stimulation of GluR2 transcription. Intracerebroventricular treatment with VEGF similarly induced GluR2 expression in the ventral spinal cord of rats and this mechanism contributes to the protective effect of VEGF on motor neurons. PMID- 19185396 TI - Fuel particles in the Chernobyl cooling pond: current state and prediction for remediation options. AB - During the coming years, a management and remediation strategy for the Chernobyl cooling pond (CP) will be implemented. Remediation options include a controlled reduction in surface water level of the cooling pond and stabilisation of exposed sediments. In terrestrial soils, fuel particles deposited during the Chernobyl accident have now almost completely disintegrated. However, in the CP sediments the majority of (90)Sr activity is still in the form of fuel particles. Due to the low dissolved oxygen concentration and high pH, dissolution of fuel particles in the CP sediments is significantly slower than in soils. After the planned cessation of water pumping from the Pripyat River to the Pond, significant areas of sediments will be drained and exposed to the air. This will significantly enhance the dissolution rate and, correspondingly, the mobility and bioavailability of radionuclides will increase with time. The rate of acidification of exposed bottom sediments was predicted on the basis of acidification of similar soils after liming. Using empirical equations relating the fuel particle dissolution rate to soil and sediment pH allowed prediction of fuel particle dissolution and (90)Sr mobilisation for different remediation scenarios. It is shown that in exposed sediments, fuel particles will be almost completely dissolved in 15-25 years, while in parts of the cooling pond which remain flooded, fuel particle dissolution will take about a century. PMID- 19185397 TI - Approaches to assessment of alcohol intake during pregnancy in Swedish maternity care--a national-based investigation into midwives' alcohol-related education, knowledge and practice. AB - OBJECTIVE: to evaluate how much education midwives in Sweden have undertaken to help them assess alcohol intake during pregnancy, and what tools they use to identify women who may be at risk of drinking during pregnancy. DESIGN: a national survey was conducted in March 2006, using a questionnaire constructed by a Swedish team of researchers and clinicians. SETTING: maternity health-care centres in Sweden. PARTICIPANT: 2106 midwives. FINDINGS: nearly all midwives stated that they had excellent or good knowledge concerning the risks associated with drinking during pregnancy. They considered themselves less knowledgeable about detecting pregnant women with risky alcohol consumption before pregnancy. The majority of the midwives had participated in some education in handling risky drinking. Almost half of the midwives assessed women's alcohol intake before pregnancy. Important facilitators for increased activity concerned recommendations and decisions at different levels (national, local and management) on how to address alcohol with expectant parents and work with risky drinkers. KEY CONCLUSIONS: more education was associated with more common use of a questionnaire for assessment of women's alcohol intake before pregnancy, and more frequent counselling when identifying a pregnant woman whose pre-pregnancy consumption was risky. PMID- 19185398 TI - Exploring the views of women on using nicotine replacement therapy in pregnancy. AB - OBJECTIVE: to explore women's views surrounding the use of nicotine replacement therapy (NRT) to aid smoking cessation. DESIGN AND SETTING: a qualitative approach using semi-structured interviews undertaken on 10 pregnant women in a semi-rural area of England. FINDINGS: the two main themes that emerged from the study were that NRT was an important component in stopping smoking, but this was in conjunction with support given by the midwife advisor. Both themes appeared to be equally important to the women in aiding their attempts at smoking cessation. Also, the importance of tailored interventions should not be dismissed when aiding women in smoking cessation. CONCLUSIONS: exploring the views of women enabled the researchers to gain a greater understanding of the problems and achievements that women encountered when using NRT as an aid to stop smoking during pregnancy. The study also demonstrated that smoking cessation strategies can be successful if they are tailored to the individual needs of each woman, taking into account her dependency, domestic circumstances and willingness to change. IMPLICATIONS FOR PRACTICE: the findings of this study add to the available research surrounding the efficacy of NRT in pregnancy. There is little evidence that the views of pregnant women have been taken into account prior to this study; as such, this report offers a different dimension to the available evidence. Although these findings could be adopted and utilized by practitioners, there is still the need for further research in this area on a larger scale. PMID- 19185399 TI - Randomized and observational studies in nephrology: how strong is the evidence? AB - The strength of the evidence that can be obtained from observational studies and randomized clinical trials (RCTs) has been widely discussed. This report critically examines 2 perspectives of this discussion in the setting of clinical studies in nephrology that have the objective of determining the efficacy of therapeutic interventions. Emphasizing the fundamental role of randomization in supporting causal inference, the first perspective asserts the strict superiority of RCTs for showing efficacy and emphasizes the susceptibility of observational studies to bias that cannot be corrected without invoking untestable assumptions. We also examine counterarguments to this strictly hierarchical perspective that support a greater role for observational research. The counterarguments cite limitations of RCTs, systematic reviews of the concordance between results of observational studies and RCTs, and methodological advances that have strengthened the inferences that can be made from observational data. Ultimately, we support an integrative approach that targets the use of observational studies and RCTs at different stages of the research process based on their respective strengths and weaknesses and seeks to maximize the information gained by joint evaluation of both types of evidence. PMID- 19185400 TI - The effect of combined calcium and vitamin D3 supplementation on serum intact parathyroid hormone in moderate CKD. AB - BACKGROUND: Studies addressing the effects of vitamin D(3) supplementation on secondary hyperparathyroidism in patients with moderate chronic kidney disease are scarce. STUDY DESIGN: Post hoc analysis of the randomized clinical trial Vitamin D, Calcium, Lyon Study II (DECALYOS II) to assess effects according to baseline estimated glomerular filtration rate (eGFR). SETTING & PARTICIPANTS: Multicenter, randomized, double-blinded, placebo-controlled study of 639 elderly women randomly assigned to calcium-vitamin D(3) fixed combination; calcium plus vitamin D(3) separate combination, or placebo. INTERVENTIONS: Placebo or calcium (1,200 mg) and vitamin D(3) (800 IU) in fixed or separate combination. OUTCOMES & MEASUREMENTS: Proportion of participants with a mean decrease in intact parathyroid hormone (iPTH) level of 30% or greater. eGFR was calculated using the 4-variable Modification of Diet in Renal Disease (MDRD) Study equation and categorized as 60 or greater, 45 to 59, and less than 45 mL/min/1.73 m(2). RESULTS: 610 participants had an eGFR at baseline: 288 (47.2%), 222 (36.4%), and 100 (16.4%) were in each decreasing eGFR category. Across decreasing eGFR groups, 88%, 86%, and 89% had 25-hydroxyvitamin D (25[OH]D) levels less than 15 ng/mL at baseline. On treatment, similar improvements in the proportion of participants achieving 25(OH)D levels greater than 30 ng/mL at 6 months were seen in all kidney function groups (43%, 49%, and 41%, respectively). Active regimens versus placebo increased mean 25(OH)D levels from baseline in all eGFR groups at all times (P < 0.001 for all). The proportion with a 30% or greater decrease in iPTH level at 6 months was 50% in all eGFR groups on treatment versus 6% to 9% for placebo (P < 0.001 for all). The effects of the intervention on iPTH levels did not differ according to baseline eGFR (interaction P > 0.1 for all times). LIMITATIONS: This study included only elderly white women. CONCLUSION: Vitamin D(3) was effective in increasing 25(OH)D and decreasing iPTH levels in patients with moderate chronic kidney disease. PMID- 19185401 TI - A forgotten cause of kidney injury in chronic myelomonocytic leukemia. PMID- 19185402 TI - Association of quarterly average achieved hematocrit with mortality in dialysis patients: a time-dependent comorbidity-adjusted model. AB - BACKGROUND: Recent publications suggest that increased mortality is associated with high hematocrit targets in erythropoietin-stimulating agent-treated patients with chronic kidney disease. We aim to further inform the debate about optimal hematocrit targets, advancing the hypothesis that the current hematocrit target may not optimize the survival of patients with end-stage renal disease. STUDY DESIGN: Cross-sectional observational study. SETTING & PARTICIPANTS: Medicare dialysis patients from 2002 to 2004 (n = 393,967). FACTORS: Quarterly average hematocrit and erythropoietin alfa (EPO) dose. OUTCOMES: Mortality hazard ratios from time-dependent Cox proportional hazard models, adjusting for comorbidities. RESULTS: N = 2,712,197 patient-facility quarters. During the study, 100,086 deaths were identified. Percentages of patient quarters within each hematocrit category: hematocrit less than 27% (2.0%), 27% to 28.49% (1.7%), 28.5% to 29.9% (2.9%), 30% to 31.49% (5.2%), 31.5% to 32.99% (9.0%), 33% to 34.49% (14.9%), 34.5% to 35.99% (19.2%), 36% to 37.49% (18.0%), 37.5% to 38.99% (12.0%), 39% to 40.49% (6.4%), 40.5% to 41.99% (3.0%), and 42% or greater (3.1%). Mortality hazard ratios from the fully adjusted model: hematocrit less than 27% (3.11), 27% to 28.49% (2.60), 28.5% to 29.9% (2.14), 30% to 31.49% (1.80), 31.5% to 32.99% (1.44), 33% to 34.49% (1.17), 34.5% to 35.99% (reference), 36% to 37.49% (0.98), 37.5% to 38.99% (1.01), 39% to 40.49% (1.13), 40.5% to 41.99% (1.32), and 42% or greater (1.57). LIMITATIONS: First, potential confounding by indication related to associations between underlying illness and mortality, anemia, and EPO responsiveness. Second, Medicare claims data reflect a range of conditions and degrees of severity not easily translated into the clinical context. Third, for Medicare claims, EPO reporting is not required if EPO is not billed. Greater than 95% of "missing hematocrit" quarters are "EPO = 0" patient quarters. Interpretation of results for the missing hematocrit and EPO = 0 use categories is complicated by data source limitations. CONCLUSIONS: We show an association between mortality and low hematocrit in dialysis patients, in part reflecting the presence of comorbidities. We also show an association between increased mortality and high hematocrit. Additional interventional trials should be undertaken to better define the optimal target for anemia management in patients with end-stage renal disease, with careful prospective identification of underlying comorbidities and clinical factors contributing to high erythropoietin stimulating agent requirement. PMID- 19185403 TI - Calcium-alkali syndrome due to vitamin D administration and magnesium oxide administration. PMID- 19185405 TI - Perceived occupational stress, affective, and physical well-being among telecommunication employees in Greece. AB - The present study examined four potential roles of work-related negative affectivity on the associations between self-reported occupational stress and physical well-being among telecommunication employees in Greece. Participants (764, predominantly male) completed a battery of self-report measures on perceived occupational stress, negative affectivity, and illness symptoms. In line with previous research, negative affectivity exerted a nuisance effect, by inflating the association between reported stressors and illness symptoms, and significantly predicted illness symptoms, over and above the effects of stressors. In addition, negative affectivity influenced reported illness symptom indirectly, through the effects of stressors, and moderated the relationship between interpersonal conflict at work and illness symptoms. The findings suggest that negative affectivity can largely explain and influence in different ways the associations between self-reported stress and physical strain. It is recommended that future studies of occupational stress should control for the effects of negative affectivity, and that health professionals should be cautious of its effects when interpreting relationships between self-reported occupational stress and physical well-being. PMID- 19185404 TI - Decreased antibody response to influenza vaccination in kidney transplant recipients: a prospective cohort study. AB - BACKGROUND: Antibody response to the inactivated influenza vaccine is not well described in kidney transplant recipients administered newer, but commonly used, immunosuppression medications. We hypothesized that kidney transplant recipient participants administered tacrolimus-based regimens would have decreased antibody response compared with healthy controls. STUDY DESIGN: Prospective cohort study of 53 kidney transplant recipients and 106 healthy control participants during the 2006-2007 influenza season. All participants received standard inactivated influenza vaccine. SETTING & PARTICIPANTS: Kidney transplant recipients administered tacrolimus-based regimens at a single academic medical center and healthy controls. PREDICTOR: Presence of kidney transplant. OUTCOMES: Proportion of participants achieving seroresponse (4-fold increase in antibody titer) and seroprotection (antibody titer > or = 1:32) 1 month after vaccination. MEASUREMENTS: Antibody titers before and 1 month after vaccination by means of hemagglutinin inhibition assays for influenza types A/H1N1, A/H3N2, and B. RESULTS: A smaller proportion of the transplantation group compared with the healthy control group developed the primary outcomes of seroresponse or seroprotection for all 3 influenza types at 1 month after vaccination. The response to influenza type A/H3N2 was statistically different; the transplantation group had 69% decreased odds of developing seroresponse (95% confidence interval, 0.16 to 0.62; P = 0.001) and 78% decreased odds of developing seroprotection (95% confidence interval, 0.09 to 0.53; P = 0.001) compared with healthy controls. When participants less than 6 months from the time of transplantation were considered, this group had a significantly decreased response to the vaccine compared with healthy controls. LIMITATIONS: Decreased sample size, potential for confounders, outcome measure used is the standard but does not give information about vaccine efficacy. CONCLUSIONS: Kidney transplant recipients, especially within 6 months of transplantation, had diminished antibody response to the 2006-2007 inactivated influenza vaccine. PMID- 19185406 TI - The developmental cognitive neuroscience of functional connectivity. AB - Developmental cognitive neuroscience is a rapidly growing field that examines the relationships between biological development and cognitive ability. In the past decade, there has been ongoing refinement of concepts and methodology related to the study of 'functional connectivity' among distributed brain regions believed to underlie cognition and behavioral control. Due to the recent availability of relatively easy-to-use tools for functional connectivity analysis, there has been a sharp upsurge of studies that seek to characterize normal and psychopathologically abnormal brain functional integration. However, relatively few studies have applied functional and effective connectivity analysis techniques to developmental cognitive neuroscience. Functional and effective connectivity analysis methods are ideally suited to advance our understanding of the neural substrates of cognitive development, particularly in understanding how and why changes in the functional 'wiring' of neural networks promotes optimal cognitive control throughout development. The purpose of this review is to summarize the central concepts, methods, and findings of functional integration neuroimaging research to discuss key questions in the field of developmental cognitive neuroscience. These ideas will be presented within a context that merges relevant concepts and proposals from different developmental theorists. The review will outline a few general predictions about likely relationships between typical 'executive' cognitive maturation and changes in brain network functional integration during adolescence. Although not exhaustive, this conceptual review also will showcase some of recent findings that have emerged to support these predictions. PMID- 19185407 TI - Response speed, contingent negative variation and P300 in Alzheimer's disease and MCI. AB - BACKGROUND: Decreased speed of information processing is a hallmark of Alzheimer's disease (AD) and mild cognitive impairment (MCI). Recent studies suggest that response speed (RS) measures are very sensitive indicators of changes in longitudinal follow-up studies. Insight into the psycho-physiological underpinnings of slowed RS can be provided by measuring the associated event related potentials (ERP). AIMS: The current study aims to investigate the relation between RS and its psycho-physiological correlates in AD and MCI. METHODS: Fifteen psychoactive drug-naive AD patients, 20 MCI patients and twenty age-matched, healthy control subjects participated. Response speed was measured during a simple (SRT) and choice reaction time task (CRT). An oddball and contingent negative variation (CNV) paradigm were used to elicit ERP. To evaluate test-retest reliability (TRR), subjects underwent a similar assessment one week after the first. RESULTS: The SRT and CRT distinguished the patient groups significantly. The P300 amplitude and latency also distinguished the groups and showed a significant correlation with response speed. The CNV amplitude did not reveal a significant difference between groups and also showed a low TRR. The TRR of the SRT, CRT and P300 amplitude and latency in general was moderate to high. The current study suggests that response speed measures on a behavioural and psycho-physiological level deserve attention as a possible marker in the diagnosis and follow-up of AD. PMID- 19185408 TI - Effects of a dental adhesive incorporating antibacterial monomer on the growth, adherence and membrane integrity of Streptococcus mutans. AB - OBJECTIVES: This study was attempted to incorporate an antibacterial monomer, methacryloxylethyl cetyl dimethyl ammonium chloride (DMAE-CB), into a commercial dental adhesive and to evaluate the antibacterial activity of the DMAE-CB incorporated adhesive after being cured against Streptococcus mutans. METHODS: DMAE-CB was incorporated at 3% (w/v) into a clinically used dental adhesive, Single Bond 2. Single Bond 2 without DMAE-CB was served as a negative control; Clearfil Protect Bond, containing an intensively researched antibacterial monomer MDPB, was enrolled as a positive control. The effects of the cured adhesives and their eluents on the growth of S. mutans were determined by film contact test and absorbance measurement, respectively. The effects of the cured adhesives on the adherence and membrane integrity of S. mutans were investigated using confocal laser scanning microscopy (CLSM) in conjunction with fluorescent indicators. RESULTS: Compared with negative control, the cured DMAE-CB-incorporated dental adhesive and positive control were found to exhibit inhibitory effect on the growth of S. mutans (P<0.05), whereas their eluents did not show detectable antibacterial activity. Moreover, the fluorescence analysis of CLSM images demonstrated that the cured DMAE-CB-incorporated adhesive and positive control could hamper the adherence of S. mutans and exert detrimental effect on bacterial membrane integrity (P<0.05). CONCLUSIONS: The incorporation of DMAE-CB can render dental adhesive with contact antibacterial activity after polymerization via influencing the growth, adherence and membrane integrity of S. mutans. PMID- 19185409 TI - Predictors for good cerebral performance among adult survivors of out-of-hospital cardiac arrest. AB - BACKGROUND: Complete neurological recovery is of great importance to survivors of cardiac arrest. Few studies have explored predictors of good cerebral performance outcomes among these. METHODS: We analyzed data from the SOS-KANTO study, a prospective, multi-center, observational study on patients who had out-of hospital cardiac arrest. We included patients with Glasgow-Pittsburgh cerebral performance categories (GP-CPC) 1 (good cerebral performance) and 2 (moderate cerebral disability) at 30 days after cardiac arrest. RESULTS: Among 122 eligible patients, 85 (70%) with GP-CPC 1 and 37 (30%) with GP-CPC 2 outcomes were analyzed. More patients with GP-CPC 1 outcome (27%) received conventional cardiopulmonary resuscitation (CPR) than those with GP-CPC 2 outcome (5%). Proportions for receiving cardiac-only resuscitation were not different between the two groups. Based on a multiple logistic-regression model constructed using age and significant variables from bivariate analyses, significant factors for GP CPC 1 outcome included: conventional bystander CPR compared to no bystander resuscitation with an odds ratio of 5.7 (95% CI, 1.1-30.4); positive pupillary reflex at the time of ED arrival with an odds ratio of 13.7 (95% CI, 3.5-53.7); spontaneous respiration at ED arrival with an odds ratio of 5.98 (95% CI, 1.6 23.0); and cardiac cause of initial arrest with an odds ratio of 5.9 (95% CI, 1.4 25.0). CONCLUSIONS: Survivors of out-of-hospital cardiac arrest with recovery to good cerebral performance were more likely to have cardiac cause of arrest and show positive pupillary reflex and spontaneous respiration at ED arrival. PMID- 19185410 TI - The need for separate analyses of survival for "in-home" arrests versus "out-of home" arrests in the determination of whether it is best to shock first or perform 3 min of CPR before the first defibrillation; in patients with ventricular fibrillation cardiac arrest when the ambulance response time is greater than 5 min. PMID- 19185411 TI - Effect of timing and duration of a single chest compression pause on short-term survival following prolonged ventricular fibrillation. AB - BACKGROUND: Pauses during chest compressions are thought to have a detrimental effect on resuscitation outcome. The Guidelines 2005 have recently eliminated the post-defibrillation pause. Previous animal studies have shown that multiple pauses of increasing duration decrease resuscitation success. We investigated the effect of varying the characteristics of a single pause near defibrillation on resuscitation outcome. METHODS: Part A: 48 swine were anesthetized, fibrillated for 7min and randomized. Chest compressions were initiated for 90s followed by defibrillation and then resumption of chest compressions. Four groups were studied-G2000: 40s pause beginning 20s before, and ending 20s after defibrillation, A1: a 20s pause just before defibrillation, A2: a 20s pause ending 30s prior to defibrillation, and group A3: a 10s pause ending 30s prior to defibrillation. Part B: 12 swine (Group B) were studied with a protocol identical to Part A but with no pause in chest compressions. Primary endpoint was survival to 4h. RESULTS: The survival rate was significantly higher for groups A1, A2, A3, and B (5/12, 7/12, 5/12, and 5/12 survived) than for the G2000 group (0/12, p<0.05). Survival did not differ significantly among groups A1, A2, A3, and B. CONCLUSIONS: These results suggest that the Guidelines 2005 recommendation to omit the post-shock pulse check and immediately resume chest compressions may be an important resuscitation protocol change. However, these results also suggest that clinical maneuvers further altering a single pre-shock chest compression pause provide no additional benefit. PMID- 19185412 TI - Influence of a lipid bilayer on the conformational behavior of amphotericin B derivatives - A molecular dynamics study. AB - Amphotericin B (AmB) is an effective but very toxic antifungal antibiotic. In our laboratory a series of AmB derivatives of improved selectivity of action was synthesized and tested. To understand molecular basis of this improvement, comparative conformational studies of amphotericin B and its two more selective derivatives were carried out in an aqueous solution and in a lipid membrane. These molecular simulation studies revealed that within a membrane environment the conformational behavior of the derivatives differs significantly from the one observed for the parent molecule. Possible reasons for such a difference are analyzed. Furthermore, we hypothesize that the observed conformational transition within the polar head of AmB derivatives may lead to destabilization of antibiotic-induced transmembrane channels. Consequently, the selective toxicity of the derivatives should increase as ergosterol-rich liquid-ordered domains are more rigid and conformationally ordered than their cholesterol-containing counterparts, and as such may better support less stable channel structure. PMID- 19185413 TI - Nonlinear processing in B-mode ultrasound affects carotid diameter assessment. AB - Noninvasive diameter assessment in the common carotid artery (CCA) by means of ultrasound is a useful technique for estimation of arterial mechanical and dynamic properties, clinical screening and treatment monitoring. Before presentation on screen, ultrasound images are subjected to nonlinear processing, e.g., logarithmic compression and noise-level thresholding, to improve visualization. In addition, signal saturation may occur, either in the received radiofrequency (RF) signals or in their envelopes. The objective of this study is to evaluate the effect of signal nonlinearities on CCA diameter measurements by means of noninvasive B-mode ultrasound, comparing the performance of two different edge detectors. In 14 healthy subjects, three repeated ultrasonic acquisitions (6 s) without saturation were performed. The acquired RF signals were subjected off-line to envelope detection, logarithmic compression and various degrees of saturation applied to the signals before or after envelope detection. For the purpose of CCA diameter estimation, artery walls were automatically outlined frame by frame. As automatic edge detectors, we considered the sustain attack filter (SAF), based on exponentially decaying reference functions, and a derivative approach (DER), relying on the positions of first derivative maxima. Both methods are applied within a region-of-interest located on the CCA. No regularization of the detected wall positions by means of pre- or postprocessing is presently applied to directly relate the outcome of the edge detectors to the applied nonlinear processing. Diameter values assessed with SAF are unaffected by logarithmic compression because of the possibility to integrate the compression characteristic of the ultrasound system into the method. The estimated diameters values obtained with DER instead show differences in the order of 10% because of compression. Saturation affects DER more than SAF; DER exhibits larger intrarecording and intrasubject variations in the estimated diameter values. Therefore, SAF gives more precise and robust CCA diameter estimates than DER, and is more suited for integration in algorithms meant for vascular ultrasound image segmentation. This study demonstrates the relevant effects of nonlinearities such as saturation and logarithmic compression on the quality of noninvasive US CCA diameter measurements. PMID- 19185414 TI - Editorial comment on: Single-port laparoscopic and robotic partial nephrectomy. PMID- 19185415 TI - Single-port laparoscopic and robotic partial nephrectomy. AB - BACKGROUND: Partial nephrectomy (PN) for small renal masses provides effective oncologic outcomes. Single-port laparoscopic (SPL) and robotic surgeries are evolving approaches to advance minimally invasive surgery. OBJECTIVE: To determine the feasibility of laparoscopic and robotic single-port PN. DESIGN, SETTING, AND PARTICIPANTS: Since 2007, evaluation of patients undergoing SPL and single-port robotic (SPR) PN at a primary referral center was performed. Patients with small, solitary, exophytic-enhancing renal masses were selected. Patients with a solitary kidney, endophytic or hilar tumors, and previous abdominal and/or kidney surgery were excluded. Perioperative and pathologic data were entered prospectively into an institutional review board (IRB)-approved database. INTERVENTIONS: Tumor location determined either an open Hasson transperitoneal or retroperitoneal approach. A single multichannel port or Triport provided intra abdominal access. The Harmonic Scalpel was used for tumor excision under normal renal perfusion. The da Vinci surgical robot was used for SPR cases. MEASUREMENTS: Patient demographics, perioperative, hematologic, and pathologic data as well as pain assessment using the Visual Analog Pain Scale (VAPS) were assessed. RESULTS AND LIMITATIONS: A total of seven patients underwent single port PN (SPL=5, SPR=2). One patient with a right anterior upper-pole mass required conversion from SPL to standard laparoscopy following tumor excision because of intraoperative bleeding. Pathology revealed six lesions compatible with renal cell carcinoma (RCC) and one benign cyst. One negative frozen section came back focally positive on final histopathology. All other surgical margins were negative. A mean difference of 3.0+/-2.0 g/dl in hemoglobin was noted in all patients. Minimal pain was noted at discharge following both laparoscopic and robotic single-port surgery (VAPS=1.7+/-1.2 vs 1+/-0.5/10). CONCLUSIONS: SPL and SPR PN is feasible for select exophytic tumors. Robotics may improve surgical capabilities during single-port surgery. PMID- 19185416 TI - White matter changes in HIV-1 infected brains: a combined gross anatomical and ultrastructural morphometric investigation of the corpus callosum. AB - OBJECTIVE: The HIV-1 associated cognitive/motor complex is characterized by cognitive, motor and behavioral disturbances. Besides a significant loss of neurons in the cerebral cortex and subcortical nuclei, a possible morphological substrate of this complex is also given by changes of the white matter as seen in HIV-1 leucoencephalopathy (HIVL), which is characterized by widespread diffuse pallor of myelin and the presence of gliomesenchymal nodules with multinucleated giant cells. METHODS: The corpus callosum as a sensitive marker for damage of the cerebral white matter was investigated by morphometry both at the macroscopic and electronmicroscopic level. RESULTS: In HIV-1 infected brains, a significant decrease of the profile area of the whole corpus callosum as well as of its different parts was noted. The absolute number of nerve fibers was significantly decreased, in particular in the frontal and occipital parts of the corpus callosum. Moreover, several morphometric parameters for nerve fibers, axons and myelin sheaths indicate in some areas a reduction of nerve fibers and axons, as well as a diminished myelin sheath thickness, whereas, in other regions, swelling of axons and myelin sheaths was observed. CONCLUSIONS: The observed changes are considered to represent subtle changes affecting nerve fibers before histological evidence of HIVL, and might represent one aspect of the morphological substrates preceeding the development of the HIV-1 related cognitive/motor complex. PMID- 19185417 TI - Early onset tension pneumocephalus following ventriculoperitoneal shunt insertion for normal pressure hydrocephalus: a case report. AB - The occurrence of tension pneumocephalus following ventriculoperitoneal (v.p.) shunt insertion is extremely rare, and is usually of delayed onset. We report a patient who developed an acute subdural tension pneumocephalus within 1 day following placement of a v.p. shunt for normal pressure hydrocephalus as a complication from shunt surgery. PMID- 19185418 TI - Low molecular weight hyaluronan inhibits colorectal carcinoma growth by decreasing tumor cell proliferation and stimulating immune response. AB - Hyaluronan modulates cancer progression by multiple mechanisms; nevertheless, its effects remain controversial. In this work, low molecular weight (LMW) hyaluronan but not high molecular weight (HMW) was found to significantly reduce colorectal carcinoma (CRC) growth in vitro and in vivo. Both survival and proliferation of CT26 tumor cells were affected by treatment with low doses of LMW HA, with involvement of Akt signaling mechanisms. We show for the first time that splenocytes isolated from LMW HA-treated animals present significantly higher proliferative capacity upon stimulation with dendritic cells (DCs) pulsed with tumor lysate. Consistently, expression of MHC class II and costimulatory molecules were increased in DCs isolated from the spleen of LMW HA-treated mice. Besides, increased tumor infiltrating lymphocytes was observed in animals treated with LMW HA. Our results suggest that LMW HA in a model of CRC triggers an activation of immune system, which is likely involved in the observed tumor growth inhibition. LMW HA is suggested as a candidate molecule for therapeutic adjuvant treatments in CRC immunotherapy. PMID- 19185419 TI - Hazardous animal waste carcasses transformation into slow release fertilizers. AB - Because of the need to address disposal of materials infected with pathogens new regulations have come into effect for the transport and disposal of dead farm animals or carcasses. For precautionary reasons, disposal to landfill, composting, biogas generation or fertilizer use are banned recycling paths because of incomplete knowledge about contamination transmission paths. Thermal treatment is recognized as a safe elimination process. Animal wastes have a high calorific value (above 16 MJ/kg). However, combustion of the organics leaves mineral residues (near 30%). The ashes contain mostly calcium and phosphate with some sodium, potassium and magnesium. We have examined the transformation of the ashes into a slow release fertilizer. We used a mixture of acids to partly dissolve the combustion residues and form slurry. In a second step, base was added to neutralize and solidify the reaction mixture. The final product was a whitish polycrystalline solid. Leaching tests were made to evaluate the nutrient release rate in laboratory columns. Water leachates were analyzed for up to ten pore-bed volumes and showed, as expected, large differences in release rates. Nitrate release was slowed and phosphate did not level even after ten pore-bed volumes. This demonstrates that insoluble precipitates (gypsum) contribute to control soluble ion release. PMID- 19185420 TI - Cadmium uptake potential of Brassica napus cocropped with Brassica parachinensis and Zea mays. AB - Cadmium uptake potential of Brassica napus co-cropped with B. parachinensis or Zea mays plants in split pot (allow the solutes to pass but prevent the interaction of roots between compartments) experiments was evaluated. Plants were grown in split pots filled with soil spiked at 0, 3, 6, 12, 25 and 50 mg Cd/kg soil. Biomass and Cd uptake were determined after 6 weeks, and rhizospheric soil solutions, extracted using soil probes, were analyzed for pH and water soluble Cd at weekly intervals. Cadmium treatments affected the biomass. Cadmium concentration in the shoots of B. napus was higher when co-cropped with B. parachinensis and significantly higher with Z. mays; however, the biomass was negatively affected implying the higher nutrient apportionment to the crop plants than B. napus. Concentration of Cd in B. napus was higher in shoots than in roots as revealed by shoot/root Cd quotient and was always >1; the quotient for B. parachinensis was approximately 1 and that of Z. mays was <1, indicating the potential of Brassicaceae members to translocate the Cd to aboveground tissue. Results indicate the feasibility of co-cropping method to clean the Cd contaminated soils. PMID- 19185421 TI - Influence of pH, soil humic/fulvic acid, ionic strength, foreign ions and addition sequences on adsorption of Pb(II) onto GMZ bentonite. AB - This work contributed to the adsorption of Pb(II) onto GMZ bentonite in the absence and presence of soil humic acid (HA)/fulvic acid (FA) using a batch technique. The influences of pH from 2 to 12, ionic strengths from 0.004M to 0.05M NaNO(3), soil HA/FA concentrations from 1.6 mg/L to 20mg/L, foreign cations (Li+, Na+, K+), anions (Cl(-), NO(3)(-)), and addition sequences on the adsorption of Pb(II) onto GMZ bentonite were tested. The adsorption isotherms of Pb(II) were determined at pH 3.6+/-0.1 and simulated with the Langmuir, Freundlich, and D-R adsorption models, respectively. The results demonstrated that the adsorption of Pb(II) onto GMZ bentonite increased with increasing pH from 2 to 6. HA was shown to enhance Pb(II) adsorption at low pH, but to reduce Pb(II) adsorption at high pH, whereas FA was shown to decrease Pb(II) adsorption at pH from 2 to 11. The results also demonstrated that the adsorption was strongly dependent on ionic strength and slightly dependent on the concentration of HA/FA. The adsorption of Pb(II) onto GMZ bentonite was dependent on foreign ions in solution. The addition sequences of bentonite/Pb(II)/HA had no effect on the adsorption of Pb(II). PMID- 19185422 TI - Reactor model development: the removal performance of ferrous-catalysed photo oxidation process by examining the reaction parameters. AB - The removal performance of the ferrous catalysed photo-oxidation process was investigated through the examination of major process parameters including pH levels and dosages of ferrous (Fe(2+)) and hydrogen peroxide (H(2)O(2)). A common used herbicide, alachlor, was used as a target compound in the degradation process. In the study, alachlor was found to be effectively degraded by hydroxyl radicals (HO) which were generated by UV/Fe(2+)/H(2)O(2) in the oxidation process. It was interesting to find that the pattern of reaction kinetics of alachlor varied depending on the initial concentrations of Fe(2+) and H(2)O(2). An optimum H(2)O(2) dosage was determined. This was practically useful because the overdose of H(2)O(2) would cause the process retardation. The conventional pseudo-first-order kinetics and two-stage first-order kinetics were observed at lower and higher Fe(2+) concentrations, respectively. Models were proposed and used to stimulate the kinetic process. Thus, design charts were established for determining the reaction time (i.e., reactor sizing) required for predetermined removal performance of alachlor under different concentrations of H(2)O(2) and Fe(2+). PMID- 19185423 TI - Behavior of dibutyl phthalate in a simulated landfill bioreactor. AB - In this study, the behavior of dibutyl phthalate (DBP) from municipal solid waste (MSW) in the leachate and refuse of two simulated landfill bioreactors was compared. In one reactor, the leachate was circulated between a landfill and a methanogenic reactor, while the other reactor was operated using direct recirculation of the leachate. The results revealed that the original concentration of DBP in the refuse was approximately 18.5 microg/g, and that this concentration decreased greatly during decomposition of the waste for both reactors. Furthermore, the major loss of DBP from the landfill occurred in an active methanogenic environment in the later period, while the environment was acidic due to a high concentration of volatile fatty acids (VFA) and contained a large volume of biologically degradable material (BDM) during the early stage. Circulating the leachate between the landfill and a methanogenic reactor resulted in an increase in the biodegradability of MSW and a high degree of waste stabilization. Furthermore, DBP degraded more rapidly in the landfill that was operated in conjunction with the methanogenic reactor when compared to the landfill in which there was direct leachate recirculation. PMID- 19185424 TI - Sorption behavior of Pb(II) and Cd(II) on iron ore slime and characterization of metal ion loaded sorbent. AB - The present investigation evaluates the sorption effectiveness of Pb(II) and Cd(II) ions on iron ore slime (IOS) obtained from Jindal Steel Ltd., Vijayanagaram, India. The sorption followed pseudo-second-order kinetics for both the cations. Pb(II) and Cd(II) sorption increased with the increase in pH from 2 to 4.5. The sorption data fitted well to Freundlich model as compared to Langmuir model. Synergistic effect of Pb(II) and Cd(II) on their sorption on IOS sample showed that Pb(II) sorption increases in presence of Cd(II) whereas Cd(II) sorption decreases. Presence of chloride or sulphate resulted in increased Pb(II) sorption but adversely affected Cd(II) sorption. The XRD patterns of Pb(II) adsorbed on IOS sample showed disappearance of some silica peaks and shifting of hematite peaks corresponding to 104 and 110 plane. For Cd(II) sorbed IOS sample, only peak shift for hematite of 104 and 110 plane was observed. Shifting of IR bands indicated that the Pb(II) sorption occurred through an inner sphere mechanism where as Cd(II) sorption occurred through outer sphere mechanism. EPMA studies showed that Pb(II) form a uniform thin layer and Cd(II) concentrate only on iron oxide phase. Regeneration and stability data on metal ion loaded IOS sample has been included. PMID- 19185425 TI - Hydrothermal processing of MSWI fly ash--towards new stable minerals and fixation of heavy metals. AB - A hydrothermal processing strategy of MSWI fly ash is presented for obtaining stable minerals with low toxic potential. Different hydrothermal conditions were tested to obtain high yields of new stable minerals. Experimental parameters including temperature, nature and molarity of alkali reagents, and reaction time were evaluated. The chemical stability of hydrothermal products was examined by the toxicity characteristic leaching procedure (TCLP) test and subsequent XRD for the leached residue. The significant amounts of Al-substituted 11A tobermorite and katoite in addition to minor amounts of zeolites were formed under experimental conditions at 0.5M NaOH, 180 degrees C for 48 h, however KOH treatment in a similar regime resulted in smaller amounts of Al-substituted 11A tobermorite and katoite. Similarly, a product of mixed Al-substituted 11A tobermorite and katoite could be formed from the washed fly ash treated in 0.5M NaOH at 180 degrees C for 48 h. Under the acidic condition, the treated fly ash exhibited an excellent stability of the mineral assemblage and less release of heavy metals relative to the untreated parent materials. PMID- 19185426 TI - Study of N-n-butyl-N-(2-nitroxyethyl)nitramine in RDX based gun propellant. AB - The current trend in the development of gun propellant is to replace the non energetic plasticizers with the energetic plasticizer. This leads to better energetic performance of gun propellant. The purpose of the present paper is to explore the possibility of enhancing the ballistic performance of RDX based gun propellant by incorporating an energetic plasticizer into the propellant composition. Compositions containing N-n-butyl-N-(2-nitroxyethyl)nitramine (Bu NENA) as an energetic plasticizer with varying percentage of RDX have been studied theoretically and experimentally. Performance in terms of ballistic parameters, sensitivity, thermal characteristics, stability and mechanical properties was evaluated and compared with the vis-a-vis compositions containing dioctylphthalate as a non-energetic plasticizer. Experimental data on comparative study indicates that Bu-NENA based propellants are superior to DOP based propellants in respect of ballistic performance. PMID- 19185427 TI - Improving the energy spread and brightness of thermal-field (Schottky) emitters with PHAST--PHoto Assisted Schottky Tip. AB - Using a relatively simple model of photoemission we derive an expression for the reduced on axis brightness of a thermal-photofield emitter. We then show that it is theoretically possible to reduce the energy spread of a Schottky (thermal field) emitter whilst increasing the reduced brightness. This can be achieved by the illumination of the tip with a high intensity laser light. We call the source PHAST-PHoto Assisted Schottky Tip. We find that due to the strong E-fields applied PHAST may operate at photon energies below the (Schottky reduced) work function. Thus removing the need for UV lasers, we will show that it is in fact preferable to work in the red, or in the green. The necessary laser intensities probably limit the application to pulsed operation. PMID- 19185428 TI - Prevalence of Cryptosporidium andersoni: a molecular epidemiological survey among cattle in India. AB - Cryptosporidiosis is an important and established cause of calfhood morbidity in bovines. The present communication reports the prevalence of Cryptosporidium infection among juvenile and adult cattle (6-24 months old) in India based on examination of faecal samples collected from 350 animals across three different agro-climatic regions of the country and further confirmation by a two-step nested PCR assay targeting 18S ssu rRNA gene. A total of 45 samples were positive for Cryptosoridium species by nested PCR assay. The PCR products were subjected to restriction fragment length polymorphism (RFLP) analysis using SspI and VspI restriction enzymes for species differentiation. The results showed that the species involved in all the samples found positive was Cryptosporidium andersoni. The overall prevalence rate was 12.85%, with highest occurrence in the northern states (14.37%) of the country. The animals between age group of 6-12 months were mostly affected (21.67%) and the season wise prevalence of infection was more during the hot and humid monsoon season (20.16%). The results clearly demonstrated that C. andersoni is the major Cryptosporidium species affecting juvenile and adult cattle in three agro-climatically different geographical regions of India. This is the first report on prevalence of C. andersoni in bovines from India the confirmation of which is based on application of nested PCR and PCR-RFLP based molecular tools. PMID- 19185429 TI - Adolescents' rationalizations to continue smoking: the role of disengagement beliefs and nicotine dependence in smoking cessation. AB - Adult smokers were found to reduce cognitive dissonance regarding their smoking behaviour by adhering to rationalizations or justifications to continue smoking, also known as disengagement beliefs. These beliefs were found to be an important barrier with regard to smoking cessation practices. Neither the occurrence of disengagement beliefs, nor its effect on motivation to quit and actual smoking cessation have been studied among adolescent smokers. Therefore, this prospective study among a sample of 363 adolescents examined the extent to which adolescents adhere to disengagement beliefs, and the relations between disengagement beliefs and adolescents' motivation to quit smoking, motivation change and smoking cessation. The association and interplay between disengagement beliefs and level of nicotine dependence was also assessed. Results showed that the degree to which adolescent smokers adhere to disengagement beliefs was similar to that of adults, if not stronger. Higher levels of dependence coincided with stronger adherence to disengagement beliefs. Further, when controlling for nicotine dependence, disengagement beliefs were strongly negatively associated with motivation to quit, but only marginally inversely associated with smoking cessation one year later. Nicotine dependence was the strongest barrier for smoking cessation at follow-up. PMID- 19185430 TI - Prenatal diagnosis and fetopathological investigation of dorsolumbosacral agenesis. AB - Sacral and lumbosacral spine agenesis, as characteristic signs of a rare congenital malformation--caudal regression syndrome--has been well described. However, dorsolumbosacral agenesis involving the lower thoracic, lumbar, and sacral vertebrae has rarely been reported, and prenatal diagnosis of this severe form has not been published yet. A 37-year-old woman (gravida 2, para 0) who had diabetes mellitus asked for termination of her pregnancy, because second trimester ultrasound screening showed dorsolumbosacral agenesis of the fetus. Fetopathological examination confirmed the prenatal diagnosis and showed that the lower seven thoracic and all lumbosacral segments were absent. The noticed small "bony" structure in the lumbar region supported the idea that caudal regression syndrome can be regarded as a "multisegmental" spinal dysgenesis that involves the caudal part of the spine. Reliable prenatal diagnosis of dorsolumbosacral agenesis is possible by second-trimester ultrasound. The prenatal sonologist should always try to look for and assess abnormalities during examinations. Emphasis should be placed especially on those types that have a higher risk of being present in the fetus because of the known risk factors in the particular pregnancy. Fetopathological examination emphasized the suggestion that segmental spinal dysgenesis and caudal regression syndrome may represent two faces of a single spectrum of segmental malformations of the spine and spinal cord. PMID- 19185431 TI - Effects of emodin on the gene expression profiling of human breast carcinoma cells. AB - BACKGROUND: The mechanism of emodin-mediated cell apoptosis has been investigated extensively in many types of human cancer cells. Our previous study demonstrated that emodin induced apoptosis through the decrease of Bcl-2/Bax ratio and the increase of cytoplasm cytochrome c concentration in human breast cancer BCap-37 cells. However, emodin's reaction to breast cancer cells remains elusive. MATERIALS AND METHODS: An apoptosis-associated cDNA microarray comprised of 458 known genes, namely, death receptors, calpains, death kinases, granzymes, DNA fragmentation proteins, caspases and Bcl-2 family, was used to determine the impact of emodin in breast cancer BCap-37 cells. Furthermore, the candidate emodin target genes were further evaluated via real-time quantitative PCR and Western blot analysis. RESULTS: We found that gene expression profiling in human breast cancer BCap-37 cells was altered when exposed to emodin. Thirty of the unique genes that were either induced or repressed in response to emodin-induced apoptosis were also identified. A follow-up study characterized p53, emodin induced gene, IGF-2, and emodin-repressed gene, and the downstream proteins were also seen as possible molecular targets of emodin. CONCLUSION: Data from this study provide novel evidence that emodin induces gene expression profiling changes, but has no effects on caspases. In addition, the p53 pathway may cooperate with the IGF-2 pathway, resulting in an emodin-induced apoptosis through disruption of the mitochondrial signaling pathway in BCap-37 cells. PMID- 19185432 TI - Characterization of Streptococcus suis isolates from the diseased pigs in China between 2003 and 2007. AB - The aim of this study was to illustrate the characteristics of 407 strains of Streptococcus suis (S. suis) isolated from diseased pigs in China. The results revealed that S. suis, with 56.6% of the Streptococci isolates, had replaced Streptococcus equi subsp. zooepidemicus as the predominant agent. Among the strains investigated, serotype 2 (43.2%) was most prevalent, followed by serotypes 3 (14.7%) and 4, 8, 5, 7, 1/2 (3.2-6.4%). Serotype 2 was more frequently isolated from swine with systemic infection, while serotype 3 was significantly associated with pneumonia. A high percentage of S. suis serotype 2 strains (54%) belonged to the genotype sly+ mrp+ epf+, which is a highly virulent strain, as confirmed by mice infection test. Biofilm producers only had lowly virulence compared to strains with the same genotype, indicating that biofilm could be associated with virulence but be not the character of virulent strains. The present study contributes to understanding the characteristics of Streptococcus suis and controlling Streptococcal disease in China. PMID- 19185433 TI - Quorum sensing in veterinary pathogens: mechanisms, clinical importance and future perspectives. AB - Under certain circumstances the individuals of a bacterial population may find advantages in acting together and making "collective decisions". This phenomenon is better known as quorum sensing. When the concentration of signal molecules produced by the surrounding bacteria exceeds a certain threshold, the bacterial population acts as a single organism, collectively expressing virulence genes, biofilm forming genes, etc. Several mechanisms of quorum sensing are discussed, each with its distinct signal molecules and respective receptors. Some of these mechanisms are restricted to sensing intraspecies signalling, but interspecies and even interkingdom signalling have also been described. Several veterinary pathogens such as Staphylococcus aureus, Staphylococcus pseudintermedius, Pseudomonas aeruginosa and Salmonella Typhimurium use quorum sensing as a means to optimize virulence gene expression and host colonization. Therefore, targeting of the QS mechanisms may provide a novel strategy for combating bacterial infections, also in veterinary medicine. PMID- 19185435 TI - Short-term preservation of canine preantral follicles: Effects of temperature, medium and time. AB - The use of the large pool of preantral follicles is a promising alternative to provide high numbers of fertilizable oocytes to reproductive biotechnology. This issue is particularly important to canids, since current rates of success of in vitro techniques using oocytes are very limited, and many species within this family are threatened by extinction. The aim of this study was to evaluate effects of temperature, medium and time on morphology and viability of canine preantral follicles during short-term preservation. Canine ovaries were cut into fragments which were incubated in 0.9% NaCl solution or in minimum essential medium (MEM) at 4, 20 or 38 degrees C for 2, 6, 12 or 24 h. Afterwards, preantral follicles were analyzed by histology, transmission electron microscopy and viability testing using trypan blue, calcein-AM and ethidium homodimer-1. Percentages of morphological normal and viable follicles were maintained similar to control (time 0 h) after incubation in 0.9% NaCl at 4 or 20 degrees C for up to 6h and at 38 degrees C for 2 h. Using MEM, such preservation was possible for 12h at 4 or 20 degrees C, and for 6h at 38 degrees C. These results indicate that preservation of canine preantral follicles might be better accomplished through hypothermic (4 or 20 degrees C) storage in MEM, which ensures maintenance of morphology and viability for up to 12h. PMID- 19185434 TI - The development of jaw motion for mastication. AB - The developmental course of early chewing has rarely been studied, although such knowledge is essential for understanding childhood feeding and swallowing disorders. The goal of this investigation was to quantitatively describe age- and consistency-related changes in jaw kinematics during early chewing development. An optical-motion tracking system was used to record jaw movements during chewing in 3-dimensions in 11 typically-developing participants longitudinally from 9-30 months of age. Age related changes in jaw movement were described for both puree and regular consistencies. The findings demonstrated that the development of rotary jaw motion, jaw motion speed, and management of consistency upgrades are protracted across the first two years of life. Young children did not differentiate their jaw closing speeds for puree and regular consistencies until 18-24 months of age, at which age the speed of movement was significantly slower for the puree than for the regular consistency. Horizontal jaw closing speed decreased significantly with age for the puree consistency, but not for the regular consistency. The emergence of a rotary chew pattern was not observed at the ages studied. PMID- 19185437 TI - Comparison of GC-MS and GC-IRD methods for the differentiation of methamphetamine and regioisomeric substances. AB - Gas chromatography-mass spectrometry (GC-MS) and gas chromatography-infrared detection (GC-IRD) methods were developed and compared for the differentiation of regioisomeric phenethylamines related to methamphetamine. There are a total of five regioisomeric phenethylamines (methamphetamine and four regioisomers) that produce essentially equivalent mass spectra. This unique set of five phenethylamines having the same molecular weight and elemental composition yield major mass spectral fragments at equivalent mass. The trifluoroacetyl derivatives of the primary and secondary amines yield characteristic individual fragment ions allowing structural differentiation among these regioisomers. The vapor phase infrared spectra generated via capillary gas chromatography differentiated among these compounds without the need for derivatization. The regioisomeric phenethylamines are well resolved by GC with the elution order generally determined by the degree of molecular linearity. PMID- 19185436 TI - Urine ratio of tetrahydrocortisol to tetrahydrodeoxycortisol to screen for the systemic administration of cortisone and hydrocortisone. AB - We use gas chromatography-mass spectrometry (GC-MS) to determine the urine peak area ratio of tetrahydrocortisol (THF) to tetrahydrodeoxycortisol (THS) in spot urine samples of eight male volunteers after a single intramuscular injection of 100 mg hydrocortisone (HC) and after a single oral administration of 10 mg HC at six different post-treatment times over 24 h with 1 week between the two treatments. Control spot urine samples were also obtained from a group of 100 volunteers of each sex for GC-MS analysis. In addition, one female volunteer was collected for GC-MS and isotope ratio mass spectrometry (IRMS) analysis after a single oral administration of 40 mg HC and 40 mg cortisone (C) at 15 and 10 different post-treatment times over 30 h, respectively. IRMS analysis focused on the acetylated derivative of 11-keto-etiocholanolone (11KE) and 11beta-hydroxy etiocholanolone (11OHE) as target metabolites, and on androsterone (A) as an endogenous reference compound (ERC) for calculating the corresponding delta(13)C (per thousand) depletion values. There was a small but significant sex-related difference for the THF/THS ratio in the control group with mean THF/THS ratio values of 10 and 13.5 for women and men, respectively. A cut-off value of 28 (mean+2 S.D.) for the THF/THS ratio offered a narrow detection window with 39% of suspicious samples after HC-oral treatment, and a wide detection window with 94% of suspicious samples after HC-intramuscular administration in men. For the woman the same cut-off value offered a wide detection window after HC and C administration with 100% and 90% of suspicious samples, respectively. On the basis of a cut-off value of 3 per thousand for the delta(13)C (per thousand) depletion, the exogenous origin was widely evidenced for at least one target compound in 93% and 80% of the HC and C samples, respectively. We conclude by discussing the predictive ability of the urine THF/THS ratio and its usefulness in pointing out suspicious samples resulting from the systemic administration of HC and C. PMID- 19185438 TI - Image quality in conventional chest radiography. Evaluation using the postprocessing tool Diamond View. AB - The objective of this work was to evaluate the influence of the postprocessing tool Diamond View (Siemens AG Medical Solutions, Germany) on image quality in conventional chest radiography. Evaluation of image quality remains a challenge in conventional radiography. Based on the European Commission quality criteria we evaluated the improvement of image quality when applying the new postprocessing tool Diamond View (Siemens AG Medical solutions, Germany) to conventional chest radiographs. Three different readers prospectively evaluated 102 digital image pairs of chest radiographs. Statistical analysis was performed with a p value <0.05 considered as significant. Images were evaluated on basis of the modified imaging Quality Criteria by the Commission of the European Communities. Each of the 11 image quality criteria was evaluated separately using a five point classification. Statistical analysis showed an overall tendency for improved image quality for Diamond View (DV) for all criteria. Significant differences could be found in most of the criteria. In conclusion DV improves image quality in conventional chest radiographs. PMID- 19185439 TI - Improving the false-negative rate of CT in acute appendicitis-Reassessment of CT images by body imaging radiologists: a blinded prospective study. AB - PURPOSE: To compare the accuracy of computed tomography (CT) analyzed by individual radiology staff members and body imaging radiologists in a non academic teaching hospital for the diagnosis of acute appendicitis. PATIENTS AND METHODS: In a prospective study 199 patients with suspected acute appendicitis were examined with unenhanced CT. CT images were pre-operatively analyzed by one of the 12 members of the radiology staff. In a later stage two body imaging radiologist reassessed all CT images without knowledge of the surgical findings and without knowledge of the primary CT diagnosis. The results, independently reported, were correlated with surgical and histopathologic findings. RESULTS: In 132 patients (66%) acute appendicitis was found at surgery, in 67 patients (34%) a normal appendix was found. The sensitivity of the primary CT analysis and of the reassessment was 76% and 88%, respectively; the specificity was 84% and 87%; the positive predictive value was 90% and 93%; the negative predictive value was 64% and 78%; and the accuracy was 78% and 87%. CONCLUSION: Reassessment of CT images for acute appendicitis by body imaging radiologists results in a significant improvement of sensitivity, negative predictive value and accuracy. To prevent false-negative interpretation of CT images in acute appendicitis the expertise of the attending radiologist should be considered. PMID- 19185440 TI - Identification of reaction products between drug substances and excipients by HPLC-SPE-NMR: ester and amide formation between citric acid and 5-aminosalicylic acid. AB - The reaction between the high-dose drug substance 5-aminosalicylic acid (5-ASA) and the excipient citric acid during storage of an experimental enema preparation has been studied and three isobaric reaction products, i.e., an ester and an amide with non-symmetrically substituted citric acid moieties and a symmetrical amide, were identified by combined use of HPLC-SPE-NMR and HPLC-MS. After storage for 1 week at 70 degrees C, approximately 5% of the 5-ASA present in the formulation was transformed into these impurities. Storage of the enema for 32 months at room temperature led to loss of approximately 10% of the original amount of 5-ASA, with the ester as the main reaction product. PMID- 19185441 TI - Chemical fingerprinting of Liuwei Dihuang Pill and simultaneous determination of its major bioactive constituents by HPLC coupled with multiple detections of DAD, ELSD and ESI-MS. AB - For quality control purpose, an approach of chemical fingerprinting of Liuwei Dihuang Pill (LDP) and simultaneous determination of its multiple bioactive components were established by using high performance liquid chromatograph (HPLC) coupled with multiple detection techniques. HPLC with ultraviolet detection (HPLC UV) was used to acquire its fingerprint, and HPLC with combined detections of diode array detector and evaporative light scattering detector (HPLC-DAD-ELSD) was performed to simultaneously determine eight bioactive constituents: including gallic acid, 5-hydroxymethyl furfural, morroniside, sweroside, loganin, paeoniflorin, paeonol and alisol B-23 acetate. The detection limits and quantification limits ranged in 0.11-1.93mug/mL and 0.38-3.85mug/mL, respectively. The validation of the proposed approach was acceptable, with 93.47 104.62% accuracy in recovery test. The intra- and inter-day precisions of the method were evaluated and were less than 3.87%, with accuracy from 95.3% to 103.4%. In addition, the mass spectrometry of the investigated major constituents was also studied. Based on the chromatographic fingerprint data, partial least square (PLS) and discriminate analysis were utilized to visualize the quality information of 60 batches of LDP, and a partial least square-discriminate analysis (PLS-DA) model was constructed with acceptable predictive performance for the discrimination of various products. The proposed approach was expected to be developed as a powerful tool for the quality control of LDP. PMID- 19185442 TI - Validated UPLC method for the fast and sensitive determination of steroid residues in support of cleaning validation in formulation area. AB - An ultra performance liquid chromatographic (UPLC) method was developed for simultaneous determination of seven steroid (dienogest, finasteride, gestodene, levonorgestrel, estradiol, ethinylestradiol, and norethisterone acetate) active pharmaceutical ingredient (API) residues. A new, generic method is presented, with which it is possible to verify the cleaning process of a steroid producing equipment line used for the production of various pharmaceuticals. The UPLC method was validated using an UPLC BEH C18 column with a particle size of 1.7 microm (50 mm x 2.1 mm) and acetonitrile-water (48:52, v/v) as mobile phase at a flow rate of 0.55 ml/min. Method development and method validation for cleaning control analysis are described. The rapid UPLC method is suitable for cleaning control assays within good manufacturing practices (GMP) of the pharmaceutical industry. PMID- 19185443 TI - Cortisol disrupts the ability of estradiol-17beta to induce the LH surge in ovariectomized ewes. AB - Stress disrupts the preovulatory luteinizing hormone (LH) surge in females, but the mechanisms are unknown. We tested the hypothesis that cortisol compromises the ability of estrogen to induce a preovulatory-like LH surge in ovariectomized ewes in both the breeding and nonbreeding season. Luteinizing hormone surges were induced in ovariectomized ewes by treatment with progesterone followed by a surge inducing estradiol-17beta (E2) stimulus using a crossover design. The experiment was replicated in the breeding and nonbreeding seasons. Cortisol reduced the incidence of LH surges irrespective of season. Cortisol increased the latency from E2 stimulus to the onset of the surge in the breeding season only and suppressed the LH surge amplitude during both seasons (P<0.01). We conclude that cortisol can interfere with the LH surge in several ways: delay, blunt, and in extreme cases prevent the E2-induced LH surge. Furthermore, the effect of cortisol to delay the E2-induced LH surge is more pronounced in the breeding season. These results show that cortisol disrupts the positive feedback effect of E2 to trigger an LH surge and suggest the involvement of multiple mechanisms. PMID- 19185444 TI - Diagnosis and multi-disciplinary management of hepatic metastases from gastrointestinal stromal tumour (GIST). AB - AIM: To explore the present application of diagnosis and management of hepatic metastases from GIST. METHODS: We performed a systematic review of the literature for studies concerning hepatic metastases from GIST. A literature search was performed using the Medline/PubMed databases to identify publications relevant to the review published from January 1998 to December 2008. Totally 113 relevant articles were retrieved. Abstracts from recent ASCO symposia were hand searched for relevant articles. After the primary filtration, articles on review and with repetitive content were excluded. The articles on clinical research, which were issued in authorized journals, were selected. At last, totally 69 articles were included for review. FINDINGS: The rate of liver metastases was reported as 15.9% in primary GISTs. The recurrence rate following surgical resection for hepatic metastases from GIST had been reported as 70-77%. For metastatic GIST patients with tyrosine kinase inhibitor (TKI) treatment, it demonstrated rates of CR, PR and SD respectively of 5.84%, 50.7%, and 32.4%. Combining repeated surgery with TKI treatment, R0/R1 resection rates range in various series between 48 and 82%. For those patients with unresectable disease confined to the liver or unable to tolerate liver resection due to co-morbidity or advanced age, RFA, HACE, TKI therapy, or even liver transplantation, can also improve survival. CONCLUSIONS: The liver is a common metastatic site for gastrointestinal stromal tumour (GIST). Appropriate initial evaluation remains paramount for selecting the correct management strategy. Multi-disciplinary management (which includes pathology, medical oncology, surgical oncology, and imaging expertise) of this disease is important for both curative and palliative treatment in these patients. Combining repeated surgery with TKI treatment may be the most effective management for GIST patients with liver metastases. PMID- 19185445 TI - [Epidemiology of chronic obstructive pulmonary disease]. AB - The chronic obstructive pulmonary disease (COPD) is responsible for important morbidity, mortality and health costs. Projection in 2020 suggests that COPD will be the 3rd cause of world mortality (6th in 1990). Although active smoking remains the main risk factor other exposure may also be involved in the development of COPD associated with genetic predisposition. The prevalence of the COPD fluctuates according to countries, age and sex. In France, mortality due to COPD remains stable in males but increases in females. A present, COPD is approximately responsible for 15,000 deaths per year. The major challenge in the near future rests on prevention and early detection of the disease. Smoking cessation allows a deceleration of the decline of the FEV1 and an improvement of total survival. PMID- 19185446 TI - [Acute exacerbations of chronic obstructive pulmonary disease]. AB - Acute exacerbation of chronic obstructive pulmonary disease (COPD) is defined by modification of the usual COPD symptoms, dyspnea, coughing and sputum, beyond daily variations, with a sudden onset, and requiring modification of the usual treatment. Exacerbations stud the course of COPD. Their frequency is variable, averaging 1-2 per year. Their frequency generally increases with COPD severity. Exacerbations impair patients' quality of life and aggravate disease prognosis by accelerating the decline in FEV1, the primary indicator of respiratory function. The most frequent causes of exacerbations are viral and bacterial respiratory infections and pollution. No cause is identified for nearly one third of all exacerbations. Most exacerbations can be treated at home, if a careful search for signs of clinical severity is negative. Treatment combines inhaled bronchodilator agents (beta-2 agonists, combined if necessary with anticholinergics) and oral corticosteroid therapy (prednisone: 0.5 mg/kg/d for 1 week) when the COPD is severe or signs of severity accompany the exacerbation. Antibiotic therapy is justified when the sputum appears purulent. Severe exacerbation may require oxygen therapy in cases of severe hypoxemia (PaO(2)<60 mm Hg) or mechanically assisted ventilation, essentially by noninvasive ventilation in cases of respiratory acidosis (pH<7.35). Noninvasive ventilation improves dyspnea and respiratory acidosis, diminishes respiratory frequency, intubation, duration of hospitalization, nosocomial infections, and mortality. Pulmonary follow-up is necessary after an exacerbation, especially to prevent the recurrence of exacerbations by measures that have been demonstrated to be effective, including help in smoking cessation, adaptation of COPD treatment, vaccination against influenza and pneumonia (pneumococci), and respiratory rehabilitation. Early diagnosis and rapid treatment of exacerbations can limit their impact, improve quality of life, and reduce the risk of hospitalization. PMID- 19185447 TI - [Vaccination in patients with cirrhosis]. AB - Patients with cirrhosis are immunocompromised and have an increased risk of infection, with a worse outcome. Some of those infections may be prevented by vaccination. Immunization can reduce the morbidity and mortality associated with cirrhosis. Immunizations against hepatitis A and B viruses, influenza and pneumococcus are recommended by the French Haute Autorite de Sante since 2007. Vaccination against hepatitis A is recommended in non-immunized cirrhotic patients. Vaccination against hepatitis B is recommended in every cirrhotic patient with no serological markers, and post-vaccinal antibodies titer should be checked. Annual influenza immunization can be done in cirrhotic patients, and pneumococcal polysaccharide vaccination should be repeated after 5 years. Few data regarding vaccination coverage are available, but studies suggest that immunization rates are too low in this population. PMID- 19185448 TI - [Social vulnerability is more frequent in victims of interpersonal violence: value of the EPICES score]. AB - OBJECTIVES: To evaluate social vulnerability of victims of interpersonal violence having consulted a service of forensic medicine with an individual index of social vulnerability. POPULATION AND METHODS: The population of victims of interpersonal violence was composed of 275 subjects having consulted the service of forensic medicine of the CHU of Saint Etienne. The social data were collected by questionnaire. Social vulnerability was measured by an individual index (EPICES) calculated on the basis of 11 weighted questions related to material and social deprivation. This population was compared with a reference population; the reference population was people, aged more than 16, living in the Rhone-Alpes region and examined in 2005 in one of the Health examination Centres (HECs) of the French General Health Insurance System, that is 7553 men and 6002 women. The comparisons between the two populations were made after redressing the population of the HECs on various socio-demographic data of the Rhone-Alpes region. The relations between violence and the variables studied were measured by odds ratios adjusted on age and sex. RESULTS: The population of the victims of violence is younger than the reference population (p<0.001). It is characterized by a lower level of education (p<0.001, 15% in the higher education level vs 23%) and the categories Employees and Manual workers are more frequent (p<0.001). The situation with respect to employment is also different between the two populations, unemployment rate is higher (OR=2.25) and the retired are fewer (OR=0.41). Subjects in social vulnerability are more frequent in the victims (57% vs 36%). All these differences persist after adjustment on age and sex. The context in which the aggression took place (family, public area or at work) varies significantly according to social vulnerability. On the other hand, the majority of the other medico-legal characteristics are not different according to the level of social vulnerability. CONCLUSION: The population of the victims of interpersonal violence has a socio-economic profile different from the reference population. Social vulnerability is associated with interpersonal violence, in particular with violence in the public and family area. PMID- 19185449 TI - How do educational attainment and gender relate to fluid intelligence, crystallized intelligence, and academic skills at ages 22-90 years? AB - Educational attainment and gender differences on fluid intelligence (Gf), crystallized intelligence (Gc), and academic skills in reading, math, and writing were analyzed for stratified adult samples ranging in age from 22 to 90 years. The data sources were the adult portions of the standardization samples of the second editions of Kaufman Brief Intelligence Test (N = 570) and the Kaufman Test of Educational Achievement-Brief Form (N = 555). Five univariate analysis of covariance were conducted with age as the covariate. Correlational analysis supplemented the covariate analyses to better understand the relationship of the five variables to education. All variables related significantly and substantially to years of formal schooling, an important finding in view of the key nature of this background variable for conducting neuropsychological assessments, as elaborated by Heaton and his colleagues. Surprisingly, Gf related just as strongly to education as did the school-related Gc. Among academic skill areas, math correlated higher with years of formal schooling than did either reading or writing. Women significantly outperformed men on the writing test and the reverse was true for the math test; other gender differences were not significant. These analyses fill a gap in the literature regarding the nature of gender and education differences in academic skills for heterogeneous samples of normal adults between young adulthood and old age and have practical implications for neuropsychological assessment. PMID- 19185450 TI - The cricothyroid joint--functional aspects with regard to different types of its structure. AB - The cricothyroid joint (CTJ) plays a key role in pitch adjustment of the human voice. It allows an external elongation of the vocal fold performed by the cricothyroid muscle with a consecutive stretching and increasing of tension. Phonosurgical methods such as cricothyroid approximation need sophisticated investigations on anatomical and functional principles because of the low satisfaction rates. Fifty cadaveric specimens were analyzed to reveal the morphological and functional anatomy of the CTJ focusing on possible gliding movements in a horizontal and vertical direction. The cartilaginous surfaces of the CTJ were categorized according to Maue and Dickson into three different types (type A: well-defined facet; type B: no definable facet; type C: flat cartilage surface or protuberance) and functional correlations examined. Side different statements and intraindividual differences between male and female specimens were included. Besides from rotational movements, the CTJ allowed horizontal and vertical gliding movements depending on the different types of the cartilaginous surfaces. Especially the difference concerning mobility between type A and the others was highly significant (P<0.001). Two thirds of our specimens showed a similar type in both CTJs, whereas in one third it was asymmetric. In comparison to the possible change of distances between horizontal gliding movements and rotation, rotation caused significant elongation of the vocal folds, which should be proposed in phonosurgical methods for cricothyroid approximation. PMID- 19185451 TI - Perceptual distances of breathy voice quality: a comparison of psychophysical methods. AB - Experiments to study voice quality have typically used rating scales or direct magnitude estimation to obtain listener judgments. Unfortunately, the data obtained using these tasks are context dependent, which makes it difficult to compare perceptual judgments of voice quality across experiments. The present experiment describes a simple matching task to quantify voice quality. The data obtained through this task were compared to perceptual judgments obtained using rating scale and direct magnitude estimation tasks to determine whether the three tasks provide equivalent perceptual distances across stimuli. Ten synthetic vowel continua that varied in terms of their aspiration noise were evaluated for breathiness using each of the three tasks. Linear and nonlinear regressions were used to compare the perceptual distances between stimuli obtained through each technique. Results show that the perceptual distances estimated from matching and direct magnitude estimation task are similar, but both differ from the rating scale task, suggesting that the matching task provides perceptual distances with ratio-level measurement properties. The matching task is advantageous for measurement of vocal quality because it provides reliable measurement with ratio level scale properties. It allows the use of a fixed reference signal for all comparisons, thus allowing researchers to directly compare findings across different experiments. PMID- 19185452 TI - Vocal tract in female registers--a dynamic real-time MRI study. AB - The area of vocal registers is still unclarified. In a previous investigation, dynamic real-time magnetic resonance imaging (MRI), which is able to produce up to 10 frames per second, was successfully applied for examinations of vocal tract modifications in register transitions in male singers. In the present study, the same MRI technique was used to study vocal tract shapes during four professional young sopranos' lower and upper register transitions. The subjects were asked to sing a scale on the vowel /a/ across their transitions. The transitions were acoustically identified by four raters. In neither of these transitions, clear vocal tract changes could be ascertained. However, substantial changes, that is, widening of the lips, opening of the jaw, elevation of the tongue dorsum, and continuous widening of the pharynx, were observed when the singers reached fundamental frequencies that were close to the frequency of the first formant of the vowel sung. These findings suggest that in these subjects register transition was not primarily the result of modifications of the vocal tract. PMID- 19185453 TI - Using electroglottographic real-time feedback to control posterior glottal adduction during phonation. AB - The goal of this pilot study was to determine whether the ability to change the degree of posterior glottal adduction (PGA) during phonation can be acquired more easily with the aid of electroglottographic (EGG) real-time feedback. The subject was a 37-year-old untrained female with habitually breathy voice. Before the experiment, she participated in one voice coaching session where exercises for increasing PGA were explained and executed. During the experiment, phonation has been monitored simultaneously with videostroboscopy, electroglottography, and audio recording. While phonating, the subject saw amplitude and period normalized EGG waveform representing one glottal cycle consecutively changing over time. The assignment was to increase the width of the EGG waveform during phonation. Laryngeal imaging revealed a posterior glottal chink during habitual phonation. The subject could only introduce intentional changes into the EGG waveform after its relevance had been explained, and after recapitulation of the exercises of the voice coaching session: An increase of the EGG waveform width coincided with the increase of high-frequency partials and an increase of PGA. For pitches B3 and B4, full glottal closure could be achieved. At G5, a reduction of the posterior glottal chink occurred. The findings of this study suggest that the skill to control the degree of PGA can be acquired, and that EGG real-time feedback can be a crucial element in optimizing the process of skill acquisition, but only if (1) the context and nature of the feedback is explained and (2) proper instructions are provided. The EGG contact quotient might not be sensitive to changes of PGA in falsetto phonation. PMID- 19185454 TI - Measuring quality of life in dysphonic patients: a systematic review of content development in patient-reported outcomes measures. AB - To review existing patient reported outcomes measures (PROMs) used in dysphonic populations to assess the procedures used in their development and the extent to which these meet current development standards for content generation and psychometric evaluation. The study is a systematic review. A systematic review of Medline, Cumulative Index to Nursing & Allied Health, and Health and Psychosocial Instruments databases was completed using voice, quality of life, and PROMs as keywords. We identified all patient or parent-reported questionnaires measuring quality of life associated with voice disorders from the review findings. Questionnaires were appraised for adherence to international guidelines for the development and evaluation of PROMs as outlined by the Scientific Advisory Committee of the Medical Outcome Trust. Nine PROMs fulfilled the inclusion criteria. The quality of these questionnaires was variable with regard to instrument development and none met all of the current, recommended criteria. Of the nine questionnaires, the Voice Symptom Scale underwent the most rigorous development process. Furthermore, many instruments have been augmented to allow for proxy administration, failing to address quality of life-related issues specific to the target population. Instrument development is often overlooked when attempting to quantify patient reported outcomes in dysphonic patients. Careful instrument development procedures are required to ensure that PROMs are valid, reliable, and responsive. Our review suggests that the deficits in psychometric properties of the current voice-related PROMs may be, at least in part, due to deficits in the development process. Furthermore, these data suggest the potential utility of a novel PROM adhering to rigorous international standards to better ensure that clinicians appreciate the variables most relevant to patients with voice disorders and address some of the psychometric shortcomings of the currently used questionnaires. PMID- 19185455 TI - Effects of training on time-varying spectral energy and sound pressure level in nine male classical singers. AB - The male classical singing voice is a musical instrument that is very important in western culture. It has many acoustic features which should change and improve over the period in which the singer trains. In this study we compare nine singers in different stages of training, from university level students through to international soloists. Typically, Energy Ratio (ER; a measure of mean spectral slope) and mean sound pressure level (SPL) may be calculated to summarize an entire singing sample. We investigate an alternative approach, by calculating the time-varying ER and SPL. The inspection of the distribution of these descriptors over an aria's time period yields a more detailed picture of the strategies for high-frequency energy production used by singers with different levels of training. PMID- 19185456 TI - Vocal fold hypomobility secondary to elective endotracheal intubation: a general surgeon's perspective. AB - This study was performed retrospectively to evaluate the incidence of documented vocal fold injury as a result of elective endotracheal intubation during general surgical procedures. Medical record review was performed at a single institution and all surgical cases reviewed which required endotracheal intubation in the nonemergent setting between April 1, 2003 and August, 31, 2007. Cases with unexpected and documented vocal fold immobility postoperatively formed the study cohort, and data were gathered regarding diagnosis and procedures performed. Of 23,010 general surgery cases performed during the study period, only seven documented cases of vocal fold paralysis were discovered (0.03%). There were five women and two men in the group; all were adults. Only one patient had a primary diagnosis related to the head and neck. Comorbidities were recorded as well, but there were no statistically significant patterns discerned. Furthermore, during the study period, a total of 31 patients overall (both surgical and nonsurgical) were admitted who carried a primary diagnosis of vocal fold paralysis. Therefore, the study cohort therefore constituted 22.6% of this total. Finally, cohort patients spent a total of 150 days in hospital during the study period; this length of stay (an average of 16.7 hospital days per patient) was significantly longer than the average of 5.1 days, presumably at least in part related to the vocal paralysis. PMID- 19185457 TI - Subjective evaluation of the long-term efficacy of speech therapy on dysfunctional dysphonia. AB - The objective of the study was to measure the subjective long-term effects of vocal therapy (more than 6 months) in 29 patients suffering from dysfunctional dysphonia. Four subjective measurements were used: one visual analog scale ranging from 0 (not efficient at all) to 100 (very efficient); one question "Is the speech therapy still efficient?," answered by yes or no; the GRB parameters of the GRBAS scale (evaluated by the patient); and the VHI-10. The results show that 76% of the patients consider that the vocal therapy they received is still efficient. On the vocal quality (G), roughness (R), and breathiness (B) parameters, we observe a high degree of satisfaction on the G parameter (median=77). The scores on the R and B parameters are lower (median R=12, S=5). Roughness is significantly correlated to the Grade (Spearman coefficient r=0.516, P=0.004), whereas breathiness is not (P=0.251, NS). The comparison of the results on the VHI-10 pre and post treatment shows a significant decrease in the grade of perceived handicap (medians 15 vs 11, P=0.017). The correlations between the judgment of the treatment's efficacy and the vocal quality is significant (r=0.623, P<0.001) as well as the correlation between the VHI-10 and the length of the treatment (r=0.416, P=0.035). The conclusion can be made that speech therapy plays an important role in long-term treatment of dysfunctional dysphonias. PMID- 19185458 TI - A study of voice changes in various phases of menstrual cycle and in postmenopausal women. AB - Larynx is responsible for the generation of voice and subsequently impacts on communication, social interaction, personality, and artistic expression. The vocal instrument is comprised of the vibratory body, the respiratory power source, and the oropharyngeal resonating chamber. The lungs are the power supply, the larynx is the vibratory source, and the supraglottal vocal tract (supraglottal pharynx oral cavity, nasal cavity) is the resonator that shapes the sound into words and songs. During the phase of expiration as the diaphragm relaxes and the chest wall recoils, air is pushed through the nearly closed vocal folds. The aerodynamic forces of the air column and myoelastic properties of the vocal folds are responsible for the repeated opening and closing of the glottal tissue that pulses that air column as it flows out. These disruptions in the steady state of tracheal air pressure by glottal activity and vocal fold vibrations result in voice production. Voice is characterized by its frequency intensity and harmonics. The harmonics are hormonally dependent. This is illustrated by changes that occur during male and female puberty. The female voice evolves from childhood to menopause under the varied influences of estrogen, progesterone, and testosterone. These hormones are the dominant factor in determining voice changes throughout life. Female voice has a fundamental frequency one-third lower than that of a child. In males, androgen released at puberty is responsible for the male vocal frequency being an octave lower than that of a child. The females have a reproductive system, which undergoes a regular cyclic change known as the menstrual cycle. Laryngeal changes are evident and fluctuate systematically during the reproductive years with the menstrual cycle. The main objective of this experiment is to provide a solid ground with evidence of changes in voice because of sexual hormones, which will form the base of a multidisciplinary approach to a comprehensive and integrated understanding of premenstrual and menopausal female voice. PMID- 19185459 TI - Flexible laryngoscopy: a comparison of fiber optic and distal chip technologies part 2: laryngopharyngeal reflux. AB - Part 1 of this paper compared fiber optic (FO) and distal chip (DC) flexible technologies in the diagnosis of vocal fold masses and mucosal wave abnormalities. Part 2 of this study was designed to evaluate the usefulness of FO and DC flexible imaging in the diagnosis of laryngopharyngeal reflux (LPR) disease. Thirty-four consecutive patients were examined with either FO or DC flexible stroboscopy followed immediately by rigid stroboscopy. Rigid stroboscopy was considered the "gold-standard" for this study. All stroboscopy segments were evaluated by two laryngologists, an otolaryngologist, a laryngology fellow, and an otolaryngology resident for physical findings of LPR using the Reflux Finding Score (RFS) and Posterior Erythema Grade (PE grade). Both flexible systems underrepresented the physical findings of LPR compared to the rigid examination, but the FO system was frequently more accurate than the DC system. For PE grade, agreement with the rigid endoscope was 95% for the FO system and 73% for the DC system. Total RFSs for both flexible systems were significantly different than RFSs from the corresponding rigid examinations (P=0.001). Raters who used the RFS more often were more consistent. More severe PE grade scores correlated well with increasing RFSs. The number of patients diagnosed with LPR (RFS>7) showed that despite differences in the category scores, the FO and DC were almost identical in how much LPR was diagnosed compared with their matched rigid examination. Because both flexible platforms significantly underrepresented reflux signs, we recommend that a rigid laryngeal telescope be used when examining the larynx for signs of LPR. If this is not available, these data suggest that a high-quality FO endoscope may be more accurate than a DC endoscope for most otolaryngologists. PMID- 19185460 TI - Sex hormones and the elderly male voice. AB - The objective was to describe influences of sex hormones on the male voice in an elderly cohort. Sixty-three elderly males were recruited to undergo assessment of voice parameters, stroboscopy, voice-related questionnaires, a blood draw, and an ultrasound examination of the laryngeal skeleton. The group was divided into men with normal hormonal status and men with lowered levels of sex hormones, called hypogonades. Depending on the level of androgens, voice parameters did not differ. In subjects with decreased levels of estrogens, a significant increase in mean fundamental frequency, as well as changes of highest and lowest frequency plus a shift of the frequency range could be detected. We could detect significant changes of voice parameters depending on status of estrogens in elderly males. Androgens appear to have no impact on the elderly male voice. To our knowledge, this is the first prospective study that correlates sex hormones with voice parameters in elderly men. PMID- 19185461 TI - The applicability of the dysphonia severity index and the voice handicap index in evaluating effects of voice therapy and phonosurgery. AB - The objective was to investigate the applicability of the Dysphonia Severity Index (DSI) and the Voice Handicap Index (VHI) in evaluating effects of intervention between groups of patients and for intrasubject differences and whether DSI and VHI are complementing measurements. Analyses of measurement data before and after intervention of 171 patients with voice disorders. The voice quality was measured objectively with the DSI. The perceived voice handicap was measured with the VHI. Three groups of patients were used: patients who had voice therapy, phonosurgery, or no intervention. DSI and VHI improved significantly after intervention in the voice therapy and the surgery group (median difference DSI 1.19 and 3.03, VHI -8 and -26, respectively). The intrasubject results were analyzed based on the test-retest variability of DSI and VHI. Significant better DSI and VHI scores after intervention were found in, respectively, 22% and 38% of the patients with voice therapy, and 56% and 78% of the patients with surgery. In the no intervention group, this was 11% and 12%. In 37% of the patients, the differences before and after intervention in DSI and VHI were in discordance. The DSI and VHI are able to show significant differences after intervention for voice disorders between groups of patients. The DSI and VHI can be used to determine a significant intrasubject result of intervention. The DSI and VHI measure each different aspects of the voice and are complementing measurements. The DSI is therefore applicable in clinical practice for objective evaluation of voice quality and the VHI for subjective evaluation of the perceived handicap by the patient self. PMID- 19185462 TI - Therapy of unilateral vocal fold paralysis with polydimethylsiloxane injection laryngoplasty: our experience. AB - The objective of this study was to document functional results and to compare objective and subjective voice measures after endoscopic laryngoplasty with injection of polydimethylsiloxane (PDMS) for the treatment of unilateral vocal fold paralysis, and to verify PDMS biocompatibility in vocal fold. The design used was a longitudinal prospective study. Fifteen patients with unilateral vocal fold paralysis underwent endoscopic injection of PDMS in general anesthesia. Accurate voice evaluation protocol (acoustic and aerodynamics analyses, GIRBAS [Grade, Instability, Roughness, Breathiness, Asthenia, and Strain] scale, videostrobolaryngoscopy, and Voice Handicap Index test) before, after surgery, and at follow-up time was performed. The median follow-up was 21.7 months (range, 6-35). Data obtained were statistically significant. All acoustic, aerodynamics, perceptive, and subjective evaluations showed a significant improvement. No complications due to PDMS were reported. Functional results were found comparable to framework surgery. Endoscopic injection laryngoplasty with PDMS is a safe and long-term option for treatment of unilateral vocal fold paralysis. PMID- 19185463 TI - Effects of cardiac resynchronization therapy on the Doppler Tei index. AB - BACKGROUND: The Tei index is an indicator of systolic and diastolic myocardial performance. We evaluated the Tei index in patients undergoing cardiac resynchronization therapy (CRT). METHODS: Forty-two patients were studied before CRT and 1 day and 6 months after CRT, comparing responders with nonresponders. RESULTS: The Tei index decreased 1 day after CRT (left ventricle [LV]: P < .001, right ventricle [RV]: P = .01) and remained lower at follow-up (LV and RV: P < .001 vs baseline). Responders had a higher LV Tei index at baseline (P = .003) and achieved a sustained improvement in Tei index at follow-up (LV: P < .001, RV: P = .002) in contrast with nonresponders (LV and RV: not significant). Baseline LV Tei index and change in LV Tei index were both correlated with LV end-systolic volume reduction after CRT (r = 0.52, P < .001, r = 0.43, P = .006). CONCLUSION: The baseline LV Tei index was significantly higher in responders and exhibited an acute and sustained improvement after CRT. The baseline RV Tei index was similar in responders and nonresponders but improved significantly only in responders. PMID- 19185464 TI - Effect of ice compression on pain after mandibular third molar surgery: a single blind, randomized controlled trial. PMID- 19185465 TI - Phase-dependent stimulation effects on bursting activity in a neural network cortical simulation. AB - PURPOSE: A neural network simulation with realistic cortical architecture has been used to study synchronized bursting as a seizure representation. This model has the property that bursting epochs arise and cease spontaneously, and bursting epochs can be induced by external stimulation. We have used this simulation to study the time-frequency properties of the evolving bursting activity, as well as effects due to network stimulation. METHODS: The model represents a cortical region of 1.6 mm x 1.6mm, and includes seven neuron classes organized by cortical layer, inhibitory or excitatory properties, and electrophysiological characteristics. There are a total of 65,536 modeled single compartment neurons that operate according to a version of Hodgkin-Huxley dynamics. The intercellular wiring is based on histological studies and our previous modeling efforts. RESULTS: The bursting phase is characterized by a flat frequency spectrum. Stimulation pulses are applied to this modeled network, with an electric field provided by a 1mm radius circular electrode represented mathematically in the simulation. A phase dependence to the post-stimulation quiescence is demonstrated, with local relative maxima in efficacy occurring before or during the network depolarization phase in the underlying activity. Brief periods of network insensitivity to stimulation are also demonstrated. The phase dependence was irregular and did not reach statistical significance when averaged over the full 2.5s of simulated bursting investigated. This result provides comparison with previous in vivo studies which have also demonstrated increased efficacy of stimulation when pulses are applied at the peak of the local field potential during cortical after discharges. The network bursting is synchronous when comparing the different neuron classes represented up to an uncertainty of 10 ms. Studies performed with an excitatory chandelier cell component demonstrated increased synchronous bursting in the model, as predicted from experimental work. CONCLUSIONS: This large-scale multi-neuron neural network simulation reproduces many aspects of evolving cortical bursting behavior as well as the timing dependent effects of electrical stimulation on that bursting. PMID- 19185466 TI - Differential relationship of frontal pole and whole brain volumetric measures with age in neuroleptic-naive schizophrenia and healthy subjects. AB - Brodmann's area (BA) 10, which occupies the frontal pole (FP) of the human brain, has been proven to play a central role in the executive control of cognitive operations. Previous in vivo morphometric studies of the FP have been limited by the lack of an accepted boundary of its posterior limit. We studied the FP gray matter volume in 23 healthy subjects who were age-, sex-, and education-matched to 23 neuroleptic-naive recent-onset schizophrenia subjects in the age span 20-40 years, using a cytoarchitectonically and functionally valid landmark-based definition of its posterior boundary that we proposed recently (John, J.P., Yashavantha, B.S., Gado, M., Veena, R., Jain, S., Ravishankar, S., Csernansky, J.G., 2007. A proposal for MRI-based parcellation of the frontal pole. Brain Struct. Funct. 212, 245-253. 2007). Additionally, we examined the relationship between FP volume and age in both healthy and schizophrenia subjects to examine evidence for a possible differential relationship between these variables across the samples. A major finding of the study was the absence of a group-level difference in frontal pole gray volumes between the healthy and schizophrenia participants. However, a more complex finding emerged in relation to age effects. The healthy participants showed an inverse relationship of FP gray volume with age, even after taking total brain volume differences into account. But this age effect was completely absent in the schizophrenia group. Moreover, all the volumetric measures in schizophrenia subjects showed substantially higher range, variance, skewness and kurtosis when compared to those of healthy subjects. These findings have implications in understanding the possible role of FP in the pathophysiology of schizophrenia. PMID- 19185467 TI - Superiority of escitalopram to paroxetine in the treatment of depression. AB - Post-hoc pooled analysis of data from two 6-month randomised controlled trials in patients with major depressive disorder (MDD) revealed superior efficacy and tolerability of escitalopram when compared with paroxetine. Escitalopram (n=394) produced a significantly (p<0.01) greater mean treatment difference of 2.0 points in primary endpoints, judged using the Montgomery-Asberg Depression Rating Scale (MADRS) total score, compared with paroxetine (n=383). Significant differences were also observed in Clinical Global Impression (CGI)--severity (escitalopram, 2.1; paroxetine, 2.4; p<0.01) and CGI--improvement (escitalopram, 1.8; paroxetine, 2.0: p<0.01). In the sub-group of severely depressed patients (baseline MADRS> or = 30), escitalopram showed further improved efficacy compared with paroxetine in all scores. This analysis supports previous observations of superior efficacy and tolerability of long-term escitalopram treatment (10 to 20 mg/day) compared with paroxetine (20 to 40 mg/day). Escitalopram is a good therapeutic option for the long-term treatment of MDD, particularly in severely depressed patients. PMID- 19185468 TI - Reduced language lateralization in first-episode schizophrenia: an fMRI index of functional asymmetry. AB - Patients with schizophrenia exhibit a decrease or loss of normal anatomical brain asymmetry that also extends to functional levels. We applied functional magnetic resonance imaging (fMRI) to investigate language lateralization in patients with schizophrenia during their first episode of illness, thus excluding effects of chronic illness and treatment. Brain regions activated during language tasks of verb generation and passive music listening were explored in 12 first-episode patients with schizophrenia and 17 healthy controls. Regions of interest corresponded to Broca's area in the inferior frontal gyrus (IFG) and Wernicke's area in the superior temporal sulcus (STS). Patients with schizophrenia had significantly smaller lateralization indices in language-related regions than controls. A similar effect was observed in their IFG and STS regions. There was no difference between the groups in the auditory cortex for the music task. Patients with schizophrenia demonstrated greater activation than the controls in temporal regions: the difference was larger in patients with more severe positive symptom subscores. In conclusion, patients with schizophrenia demonstrated loss of normal functional brain asymmetry, as reflected in diminished lateralization of language-related activation in frontal and temporal regions. This phenomenon was already present during their first episode of psychosis, possibly reflecting developmental brain abnormalities of the illness. PMID- 19185469 TI - In-situ NMR studies of isobutane activation and exchange in zeolite beta. AB - (1)H solid-state NMR techniques have been used to simultaneously detect the reactivity of both catalyst and alkane reactant protons in an in-situ experimental design. Specifically, the activation of isobutane C-H bonds by the solid acid zeolite H-Beta is directly observed while the reaction is in progress, and the rate of proton transfer between the solid catalyst surface and gaseous isobutane is quantitatively measured using isotopic (1)H/(2)H exchange methods. Arrhenius analysis of isothermal kinetic runs revealed an apparent activation barrier of 70kJ/mole for the exchange process between isobutane and the 12 membered ring H-Beta, which exceeds our previously determined value of 57kJ/mole for isobutane in the 10-membered ring H-ZSM-5 (JACS 2006, v. 128, p. 1848). Estimation of true activation energies using heat of adsorption data from the literature combined with the experimentally measured apparent E(a) suggests that the true activation barrier differs by only 6-7kJ/mole in the two catalysts. We discuss the possibility that subtle shape selectivity, or inverse shape selectivity, and lattice solvation differences between the two catalysts account for the enhanced solvation of the isobutane transition state in HZSM-5 compared to the larger channel H-Beta. In all experiments, the isobutane reagent was treated to eliminate any unsaturated impurities that might serve as initiators for carbenium-ion mechanisms, and the active catalyst was free of any organic contaminants that might serve as a source of unsaturated initiators. PMID- 19185470 TI - Theoretical investigation on the reaction of adhesion unit dopa in mussel with electrolyzing seawater. AB - Adhesive proteins secreted by the marine mussel could bind strongly to all kinds of surfaces, for instance, ship hulls and petroleum pipelines. Studies indicated that there was an unusual amino acid 3,4-dihydroxy-l-phenylanine (dopa), which was the crucial super adhesive unit in the proteins. The technology of electrolyzing seawater was employed to generate HOCl solution to hinder the adhesion. However, the detailed anti-fouling mechanism of HOCl solution remained unknown to be fully explained. Herein, we theoretically reported a study of single molecular (dopa) reaction under the HOCl solution environment, which would be helpful to reveal the anti-fouling mechanism through electrolyzing seawater. By using the density functional theory (DFT) quantum mechanics procedure, we theoretically studied the reaction mechanism of the adhesive unit dopa in mussel with electrolyzing seawater. Two possible pathways (1 and 2) were obtained (Fig. 6). The transition state for each pathway was determined, the intrinsic reaction coordinate (IRC) was analyzed and the mechanism had been confirmed. The calculations indicated dopa tended to have electrophonic attacking substitution reaction to generate 3-chlorine-4,5-dihydroxyphenylalanine (dopa-Cl) with different pathways, which hindered the formulation of conjuncted dopa-dopa and thus the stickiness among mussel adhesive proteins reduced. The transition states computation showed that pathway (1) had one transition state (TS1-1) with an activation energy of 102.22 kJ mol(-1), while pathway (2) had two transition states (TS2-1, TS2-2) with activation energies of 191.98 kJ mol(-1) and 42.00 kJ mol(-1) respectively and one intermediate (IM2-1). Rate constant value of pathway (1) was much bigger than that of pathway (2) regardless of high or low temperature, which meant that in the reaction process, pathway (1) was the favorable reaction step; but as the temperature rose, the competitiveness of pathway (2) gradually increased. After the theoretical calculation, we found that it was Cl(+) played an important and direct role in the dopa's modification. PMID- 19185471 TI - Surface modification of P(EMA-co-HEA)/SiO2 nanohybrids for faster hydroxyapatite deposition in simulated body fluid? AB - P(EMA-co-HEA)/SiO(2) nanocomposites with 0, 15 and 30 wt% of silica were obtained by copolymerization of ethyl methacrylate, EMA, and hydroxyethyl acrylate, HEA, during the simultaneous acid-catalyzed sol-gel polymerization of tetraethoxysilane, TEOS. A surface modification treatment was applied in order to reduce the induction time for hydroxyapatite (HAp) nucleation, combining a previous NaOH attack to increase the number of surface nucleating sites, and an alternate soaking process in Ca and P solutions to form apatite precursors, prior to the immersion in a simulated body fluid (SBF). The NaOH treatment was not effective by itself in shortening the HAp induction time. It introduced sodium carboxylates in the copolymer but hydrolyzed the silica network excessively, thus reducing the surface nucleating potential of its boundary silanols. Therefore, bioactivity was only due to the surface carboxylate groups of the organic phase. Maybe a controlled dissolution extent of the silica network so as to improve bioactivity could be attained by reducing the duration of the NaOH-treatment. This would be interesting in the hybrid with 30wt% of silica, because its dense silica network is not able to hydrolyze in SBF without any previous treatment, whereas the silica network in the hybrid with 15wt% of silica hydrolyzes at the surface promoting the deposition of HAp. The CaP treatment was able to coat the surfaces of the samples with a calcium phosphate layer within minutes. This amorphous calcium phosphate acted as HAp precursor, skipping the induction period in SBF. PMID- 19185472 TI - Surface chemistry and photophysical properties of a diacetylene-peptide derivative capped quantum dots Langmuir monolayer. AB - A diacetylene derivative, 10,12-pentacosadiynoic acid (PDA), was conjugated to a small peptide chain Cysteine-Cysteine-Glycine (CCG) through the solid-phase peptide synthesis. The (CdSe)ZnS core-shell quantum dots (QDs) capped with trioctylphosphine ligands were modified through a surface ligand reaction to prepare the PDA-CCG QDs conjugate. Both systems, PDA-CCG and PDA-CCG QDs, were investigated as Langmuir monolayer at the air-water interface through the surface pressure-area (pi-A) isotherms, compression-decompression cycles, stability measurements, and in situ UV-vis and fluorescence spectroscopy. Two different pi A isotherms were observed for the systems investigated showing the importance of the peptide moiety in PDA-CCG to form a Langmuir monolayer up to a surface pressure of 50 m Nm(-1) compared with 15 m Nm(-1) for the PDA component alone. The compression-decompression cycles and stability measurements for both systems suggest the formation of a stable Langmuir monolayer over 1h time period. Although the in situ UV-vis spectroscopy of PDAA-CCG and PDA-CCG QDs does not show an absorption spectrum, we observed by in situ fluorescence spectroscopy the photoluminescence (PL) of the PDA-CCG QDs at 560 nm, with an intensity of the PL increasing linearly with the increase of the surface pressure. Irradiating the PDA-CCG QDs Langmuir monolayer at 254 nm, we observe the photopolymerization with two distinct bands at 575 (blue band) and 630 nm (red band) of the polymer. PMID- 19185473 TI - A small-angle neutron scattering and rheology study of the composite of chitosan and gelatin. AB - The composite chitosan/gelatin solutions and films formed from these solutions were studied by rheological measurements, SANS and tensile tests. The relationship between the inter-molecule interactions with microstructure, rheological behaviour of a solution and eventually the mechanical performance of formed films was established. It was found that the complex formed between chitosan and gelatin was mainly through hydrogen bond but the size of the structure was also affected by electrostatic repulsions. The local structure (correlation length) and the global structure (large inhomogeneous structure size) in the composite solutions were found to be highly correlated to each other. It was also found that the interactions between these two polymers in solution were closely related to the mechanical properties of the formed films. This work will enable one to design films with desired mechanical properties through the combination of different polymers at optimum weight ratios. PMID- 19185474 TI - Strategic approaches for improving entrapment of hydrophilic peptide drugs by lipid nanoparticles. AB - In order to introduce hydrophilic peptide drugs into solid lipid nanoparticles (SLN), a technique of combining hydrophobic ion pairing (HIP) and non-aqueous oil in-oil (O/O) emulsion-evaporation was developed. Leuprolide (LR) was selected as the model drug, while sodium stearate (SA-Na) was used as the negative charged ion pairing material. The formation of leuprolide-sodium stearate (LR-SA-Na) complex was confirmed by differential scanning calorimetry (DSC). It was observed that when the molar ratio of SA-Na/LR reached 2/1, ca 88.5% LR was incorporated into the hydrophobic ion complexes with SA-Na. Compared with the conventional method of solvent diffusion in an aqueous system, the efficiency of LR drug entrapment with SLN increased from 28.0% to 74.6% by the combined technique of HIP and O/O emulsion-evaporation. In vitro drug release tests revealed that employing technique of HIP obviously reduced the burst release and slowed down the rate of drug release. At meanwhile, applying the method of non-aqueous O/O emulsion-evaporation, the longer time of drug release but relatively higher drug burst release ratio was observed in comparison with those by the solvent diffusion method in an aqueous system. The drug entrapment and release behaviors of LR-SA-Na SLN prepared by the O/O emulsion-evaporation method suggested that it could potentially be exploited as an oral delivery system for leuprolide. PMID- 19185475 TI - Predicting the probability of survival in intensive care unit patients from a small number of variables and training examples. AB - OBJECTIVE: Survival probability predictions in critically ill patients are mainly used to measure the efficacy of intensive care unit (ICU) treatment. The available models are functions induced from data on thousands of patients. Eventually, some of the variables used for these purposes are not part of the clinical routine, and may not be registered in some patients. In this paper, we propose a new method to build scoring functions able to make reliable predictions, though functions whose induction only requires records from a small set of patients described by a few variables. METHODS: We present a learning method based on the use of support vector machines (SVM), and a detailed study of its prediction performance, in different contexts, of groups of variables defined according to the source of information: monitoring devices, laboratory findings, and demographic and diagnostic features. RESULTS: We employed a data set collected in general ICUs at 10 units of hospitals in Spain, 6 of which include coronary patients, while the other 4 do not treat coronary diseases. The total number of patients considered in our study was 2501, 19.83% of whom did not survive. Using these data, we report a comparison between the SVM method proposed here with other approaches based on logistic regression (LR), including a second level recalibration of release III of the acute physiology and chronic health evaluation (APACHE, a scoring system commonly used in ICUs) induced from the available data. The SVM method significantly outperforms them all from a statistical point of view. Comparison with the commercial version of APACHE III shows that the SVM scores are slightly better when working with data sets of more than 500 patients. CONCLUSIONS: From a practical point of view, the implications of the research reported here may be helpful to address the construction of cheap and reliable prediction systems in accordance with the peculiarities of ICUs and kinds of patients. PMID- 19185476 TI - Obesity markers and estimated 10-year fatal cardiovascular risk in Switzerland. AB - BACKGROUND AND AIM: There is an ongoing debate on which obesity marker better predicts cardiovascular disease (CVD). In this study, the relationships between obesity markers and high (>5%) 10-year risk of fatal CVD were assessed. METHODS AND RESULTS: A cross-sectional study was conducted including 3047 women and 2689 men aged 35-75 years. Body fat percentage was assessed by tetrapolar bioimpedance. CVD risk was assessed using the SCORE risk function and gender- and age-specific cut points for body fat were derived. The diagnostic accuracy of each obesity marker was evaluated through receiver operating characteristics (ROC) analysis. In men, body fat presented a higher correlation (r=0.31) with 10 year CVD risk than waist/hip ratio (WHR, r=0.22), waist (r=0.22) or BMI (r=0.19); the corresponding values in women were 0.18, 0.15, 0.11 and 0.05, respectively (all p<0.05). In both genders, body fat showed the highest area under the ROC curve (AUC): in men, the AUC (95% confidence interval) were 76.0 (73.8-78.2), 67.3 (64.6-69.9), 65.8 (63.1-68.5) and 60.6 (57.9-63.5) for body fat, WHR, waist and BMI, respectively. In women, the corresponding values were 72.3 (69.2-75.3), 66.6 (63.1-70.2), 64.1 (60.6-67.6) and 58.8 (55.2-62.4). The use of the body fat percentage criterion enabled the capture of three times more subjects with high CVD risk than the BMI criterion, and almost twice as much as the WHR criterion. CONCLUSION: Obesity defined by body fat percentage is more related with 10-year risk of fatal CVD than obesity markers based on WHR, waist or BMI. PMID- 19185477 TI - Twenty-one year tracking of serum non-cholesterol sterols. The Cardiovascular Risk in Young Finns study. AB - BACKGROUND AND AIM: To show tracking of cholesterol metabolism, the ratios to cholesterol of e.g. serum cholestenol, desmosterol, and lathosterol, reflecting cholesterol synthesis, and cholestanol, campesterol, avenasterol and sitosterol, reflecting cholesterol absorption, were measured 21 years apart. METHODS AND RESULTS: In random population samples initially comprising 12- (n=162), 15- (n=158), and 18-year-old (n=148) males participating in the Cardiovascular Risk in Young Finns Study, serum sterols and squalene were measured with gas-liquid chromatography in 1980 and 2001. Quartiles of cholestanol, indicating low to high cholesterol absorption, were defined from the cholestanol values in 1980. Serum cholesterol increased in the oldest age group only, but synthesis markers (except desmosterol) increased in all age groups after the follow-up (e.g. lathosterol, total population +47.3+/-2.6% (SE), P<0.001). Campesterol (+69.0+/-3.0%, P<0.001) and sitosterol increased, avenasterol was unchanged, and cholestanol decreased ( 6.2+/-0.7%, P<0.001), respectively. The 1980 synthesis and absorption markers were interrelated with respective values 21 years later in all age groups and quartiles (e.g. lathosterol, total population 1980 vs. 2001 r=0.460, cholestanol 1980 vs. 2001 r=0.593, P<0.001 for both). Synthesis markers were highest in the first and lowest in the fourth quartile both in 1980 and 2001 (e.g. 2001, desmosterol, quartile 1, 99+/-9, quartile 4, 83+/-2 microg/mg of cholesterol, P<0.05). CONCLUSIONS: Cholesterol metabolism is significantly tracked in adolescent males over the follow-up of 21 years. Thus, high cholesterol synthesis and low absorption characterize subjects with the lowest cholestanol quartile, while those with the highest quartile have low synthesis and high absorption in both adolescence and later in young adult life. PMID- 19185478 TI - Some biological effects of scorpion envenomation in late pregnant rats. AB - Scorpion envenoming is less studied during gestation; however, it may induce various biological disturbances in maternal organism and hypothetical ones on their fetuses. The scope of this report was to elucidate some biological effects of such poisoning in late pregnant rats. Hence, TBARS levels in maternal lung, placental and fetal pulmonary and hepatic tissues and dam's biochemical blood parameters (glucose, creatinine, 17-beta estradiol, progesterone, blood nitrogen urea, sodium and potassium maternal plasma concentrations) had been evaluated after saline (G1), and scorpion venom (G2: 30 min and G3: 60 min) injections in 22nd day pregnant rats. Histological microscopic examination of these tissues was also carried out in HE-stained paraffin sections. In addition, the mean arterial blood pressure following the envenomation variations was measured in three rats from the same pool. Our results showed that Buthus occitanus tunetanus crude venom induced significant increase in maternal, placental and fetal tissues lipid peroxidation, concomitant with blood pressure elevation. Maternal plasma creatinine, estradiol and progesterone concentrations levelled up significantly after 30 min or later (60 min) after the venom injection. Except for a probable pronounced oedema and few congestions in maternal lungs and degenerative aspects of trophoblast cells, all examined tissues showed a conserved structure. These results suggest that scorpion envenomation may induce gestation process disturbances and threatens both mother's and fetus' well-being. PMID- 19185479 TI - Correcting aberrant kinetochore microtubule attachments: an Aurora B-centric view. AB - The directed movement of chromosomes during mitosis and meiosis relies on microtubule-mediated connections between spindle poles and kinetochores assembled on chromosomes. The molecular basis for the dynamic interaction between microtubules and kinetochores is just beginning to be unveiled. Here, focusing on the mitotic centromere kinase Aurora B, we review our current understanding of the signaling pathways that correct erroneous microtubule attachment at kinetochores. We evaluate several potential models that may explain how maloriented attachments are recognized and processed by the Aurora B pathway. PMID- 19185480 TI - Probing the macromolecular organization of cells by electron tomography. AB - A major goal in cell biology is to understand the functional organization of macromolecular complexes in vivo. Electron microscopy is helping cell biologists to achieve this goal, thanks to its ability to resolve structural details in the nanometer range. While issues related to specimen preparation, imaging, and image interpretation make this approach to cell architecture difficult, recent improvements in methods, equipment, and software have facilitated the study of both important macromolecular complexes and comparatively large volumes from cellular specimens. Here, we describe recent progress in electron microscopy of cells and the ways in which the relevant methodologies are helping to elucidate cell architecture. PMID- 19185481 TI - Cell structure and dynamics. PMID- 19185482 TI - The diffusive interaction of microtubule binding proteins. AB - Microtubule-based motility is often thought of as specifically referring to the directed stepping of microtubule-based motors such as kinesin or dynein. However, microtubule lattice diffusion (also known as diffusional motility) provides a second mode of transport that is shared by a much broader class of microtubule binding proteins. Microtubule lattice diffusion offers distinct advantages as a transport mechanism including speed, bidirectional microtubule end targeting, and no requirement for direct chemical energy (i.e. ATP). It remains to be seen whether a universal binding mechanism for this interaction will be identified but electrostatic interactions appear to play a significant role. In the meantime, the well-studied subject of DNA binding proteins that diffuse along the DNA backbone provides an insightful analog for understanding the nature of microtubule-based diffusional motility. PMID- 19185483 TI - Dopamine sensitized nanoporous TiO2 film on electrodes: photoelectrochemical sensing of NADH under visible irradiation. AB - Dopamine-coordinated photoactive TiO(2) nanoporous films with a wide excitation range of light in the visible region (up to 580 nm) were prepared and used for sensitive detection of NADH. Colloidal TiO(2) was firstly covered on an indium tin oxide (ITO) electrode surface and sintered at 450 degrees C to form a nanoporous TiO(2) film, then the electrode was dipped in a dopamine solution to form a dopamine-TiO(2) charge transfer complex via coordinating dopamine with undercoordinated titanium atoms on the electrode surface. This charge transfer complex provided an anodic photocurrent under visible light and the photocurrent could be largely enhanced by NADH. The photocurrent enhancement might be due to the electron transfer between NADH and the holes localized on dopamine. A new photoelectrochemical methodology for sensitive detection of NADH at a relatively low potential was developed. The detection limit of NADH was 1.4x10(-7) M, and the detection range could extend up to 1.2x10(-4) M. The dopamine-TiO(2) modified electrode exhibits its major advantages such as effective electronic transducer, fast response and easy fabrication for photoelectrochemical determination of NADH. This strategy largely reduces the destructive effect of UV light and the photogenerated holes of illuminated TiO(2) to biomolecules and opens a new avenue for the applications of TiO(2) in photoelectrochemical biosensing. PMID- 19185484 TI - Cathodic detection of H2O2 based on nanopyramidal gold surface with enhanced electron transfer of myoglobin. AB - Direct and reversible electron transfer of myoglobin (Mb), for the first time, is achieved at nanopyramidal gold surface, which was fabricated by one-step electrodeposition, with redox formal potential of 0.21+/-0.01 V (vs. Ag/AgCl) and an apparent heterogeneous electron-transfer rate constant (k(s)) of 1.6+/-0.2 s( 1). Electrochemical investigation indicates that Mb is stably confined on the nanopyramidal gold surface and maintains electrocatalytic activity toward hydrogen peroxide (H(2)O(2)). The facilitated electron transfer combined with the intrinsic catalytical activity of Mb substantially construct the third-generation biosensor for H(2)O(2). The positive redox potential of Mb at the nanostructured gold electrode gives a strong basis for determination of H(2)O(2) with high selectivity. Besides this advantage, the present biosensor also exhibits quick response time, broad linear range, and good sensitivity. The dynamic detection linear range is from 1 microM to 1.4 mM with a detection limit of 0.5 microM at 3sigma. The striking analytical performance of the present biosensor, as well as the biocompatibility of gold nanostructures provided a potential for continuous, on-line detection of H(2)O(2) in the biological system. PMID- 19185486 TI - Structure-activity studies of echinomycin antibiotics against drug-resistant and biofilm-forming Staphylococcus aureus and Enterococcus faecalis. AB - Four echinomycin antibiotics were isolated from the culture broth of a marine streptomycete, and their structures were determined by a combination of chemical and spectroscopic analyses. Antibiotic activities were measured against drug resistant and biofilm-forming strains of Staphylococcus aureus and Enterococcus faecalis. Minimum inhibitory concentrations ranging from 0.01 microM to greater than 14 microM clearly defined structure-activity relationships for antibiotic potency. Echinomycin was the most active compound with a MIC of 0.03 microM against methicillin-resistant S. aureus and 0.01 microM against biofilm-forming E. faecalis. PMID- 19185485 TI - PI 3-kinase and cancer: changing accents. AB - Research on PI 3-kinase (PI3K) is undergoing significant shifts in emphasis. Questions that have been dormant for some time are coming to the forefront, such as the relationship of PTEN to PI3K and the role of AKT in PI3K-driven oncogenesis. Two non-alpha isoforms of Class I PI3K are now established as important determinants in cancer: p110beta and p110delta. The oncogenic activities of p110beta include a non-catalytic function, a finding that will have immediate consequences for drug development. PMID- 19185487 TI - A heterocyclic molecule with significant activity against dengue virus. AB - There are no specific approved drugs or vaccines for the treatment or prevention of infectious dengue virus and there are very few compounds known that inhibit the replication of this virus. This letter describes the concise synthesis of two uracil-based multifunctional compounds. One of these compounds (1) has strong activity against dengue virus. It also exhibits low activity against a few other RNA viruses, but is highly active against yellow fever virus, a related flavivirus. It is likely that the mechanism of action of the antiviral activity of this compound is through its inhibition of the enzyme, inosine monophosphate dehydrogenase (IMPDH). Molecular modeling studies reveal that the compound can have specific hydrogen bonding interactions with a number of amino acids in the active site of IMPDH, a stacking interaction with the bound natural substrate, IMP, and the ability to interfere with the binding of NAD(+) with IMPDH, prior to the hydration step. PMID- 19185488 TI - Optimization of novel di-substituted cyclohexylbenzamide derivatives as potent 11 beta-HSD1 inhibitors. AB - Novel 4,4-disubstituted cyclohexylbenzamide inhibitors of 11beta-HSD1 were optimized to account for liabilities relating to in vitro pharmacokinetics, cytotoxicity and protein-related shifts in potency. A representative compound showing favorable in vivo pharmacokinetics was found to be an efficacious inhibitor of 11beta-HSD1 in a rat pharmacodynamic model (ED(50)=10mg/kg). PMID- 19185489 TI - Molecular hybridization, synthesis, and biological evaluation of novel chroman I(Kr) and I(Ks) dual blockers. AB - The combination of I(Kr) and I(Ks) blockade could lead to synergistic and safe class III anti-arrhythmic effect with the enhanced efficacy and reduced risk. On the rationale of structural hybridization of azimilide and HMR-1556, a novel series of I(Kr) and I(Ks) dual blockers were designed, synthesized and evaluated in vitro. One compound, 3r (CPUY11018), deserves further evaluation for its potent anti-arrhythmic activity and favorable cardiovascular profile. PMID- 19185490 TI - 1-Amido-1-phenyl-3-piperidinylbutanes--CCR5 antagonists for the treatment of HIV: part 2. AB - Optimisation of a series of 4-piperidinyltriazoles led to the identification of compound 28a which showed good whole cell antiviral activity, excellent selectivity over the hERG ion channel and complete oral absorption. PMID- 19185491 TI - Analysis of growth factor expression in affected and unaffected muscles of oculo pharyngeal muscular dystrophy (OPMD) patients: a pilot study. AB - Oculo-pharyngeal muscular dystrophy (OPMD) is characterised by progressive eyelid drooping (ptosis) and difficulties with swallowing (dysphagia). In order to determine the role of growth factors, cytokines and chemokines in the physiopathology of muscle disease we have compared the level of expression of 174 factors in both the affected (cricopharyngeal) and non-affected (sternocleidomastoid) muscles of 8 OPMD patients by means of antibody arrays. Despite an important inter-individual variability the expression of sixty factors was found to be persistently different between affected and non-affected muscles. Many of the differentially expressed factors in our study are known to be involved in the formation of fibrosis in both the liver and the lung, indicating the possibility that treatments such as those used in hepatic fibrosis may have a beneficial effect in OPMD patients. PMID- 19185492 TI - Light-dependent interactions between the Drosophila circadian clock factors cryptochrome, jetlag, and timeless. AB - Circadian clocks regulate daily fluctuations of many physiological and behavioral aspects in life. They are synchronized with the environment via light or temperature cycles [1]. Natural fluctuations of the day length (photoperiod) and temperature necessitate a daily reset of the circadian clock on the molecular level. In Drosophila, the blue-light photoreceptor Cryptochrome (Cry) mediates a rapid light-dependent degradation of the clock protein Timeless (Tim) via the F box protein Jetlag (Jet) and the proteasome, which initiates the resetting of the molecular clock [2, 3]. Cry is also degraded in the light but whereas the degradation of Tim is well characterized [4-8], the mechanism for light-dependent degradation of Cry is mostly unknown. Until now it was believed that these two degradation pathways are distinct [4, 9]. Here we reveal that Jetlag also interacts with Cry in a light-dependent manner. After illumination, Jetlag induces massive degradation of Cry, which can be prevented in vitro and in vivo by adding Tim as an antagonist. We show that the affinity of Tim for Cry and Jetlag determines the sequential order of Tim and Cry degradation and thus reveal an intimate connection between the light-dependent degradation of these two proteins by the same proteasomal pathway. PMID- 19185493 TI - Local cortical tension by myosin II guides 3D endothelial cell branching. AB - A key feature of angiogenesis is directional control of endothelial cell (EC) morphogenesis and movement [1]. During angiogenic sprouting, endothelial "tip cells" directionally branch from existing vessels in response to biochemical cues such as VEGF or hypoxia and migrate and invade the surrounding extracellular matrix (ECM) in a process that requires ECM remodeling by matrix metalloproteases (MMPs) [2-4]. Tip EC branching is mediated by directional protrusion of subcellular pseudopodial branches [5, 6]. Here, we seek to understand how EC pseudopodial branching is locally regulated to directionally guide angiogenesis. We develop an in vitro 3D EC model system in which migrating ECs display branched pseudopodia morphodynamics similar to those in living zebrafish. Using this system, we find that ECM stiffness and ROCK-mediated myosin II activity inhibit EC pseudopodial branch initiation. Myosin II is dynamically localized to the EC cortex and is partially released under conditions that promote branching. Local depletion of cortical myosin II precedes branch initiation, and initiation can be induced by local inhibition of myosin II activity. Thus, local downregulation of myosin II cortical contraction allows pseudopodium initiation to mediate EC branching and hence guide directional migration and angiogenesis. PMID- 19185495 TI - Strategic variation in mobbing as a front line of defense against brood parasitism. AB - Coevolutionary arms races, where adaptations in one party select for counter adaptations in another and vice versa, are fundamental to interactions between organisms and their predators, pathogens, and parasites [1]. Avian brood parasites and their hosts have emerged as model systems for studying such reciprocal coevolutionary processes [2, 3]. For example, hosts have evolved changes in egg appearance and rejection of foreign eggs in response to brood parasitism from cuckoos, and cuckoos have evolved host-egg mimicry as a counter response [4-6]. However, the host's front line of defense is protecting the nest from being parasitized in the first place [7-10], yet little is known about the effectiveness of nest defense as an antiparasite adaptation, and its coevolutionary significance remains poorly understood [10]. Here we show first that mobbing of common cuckoos Cuculus canorus by reed warblers Acrocephalus scirpaceus is an effective defense against parasitism. Second, mobbing of cuckoos is a phenotypically plastic trait that is modified strategically according to local parasitism risk. This supports the view that hosts use a "defense in-depth strategy," with successive flexible lines of defense that coevolve with corresponding offensive lines of the parasite. This highlights the need for more holistic research into the coevolutionary consequences when multiple adaptations and counter-adaptations evolve in concert [11]. PMID- 19185494 TI - Motor- and tail-dependent targeting of dynein to microtubule plus ends and the cell cortex. AB - BACKGROUND: Cytoplasmic dynein mediates spindle positioning in budding yeast by powering sliding of microtubules along the cell cortex. Although previous studies have demonstrated cortical and plus-end targeting of dynein heavy chain (Dyn1/HC), the regulation of its recruitment to these sites remains elusive. RESULTS: Here we show that separate domains of Dyn1/HC confer differential localization to the dynein complex. The N-terminal tail domain targets Dyn1/HC to cortical Num1 receptor sites, whereas the C-terminal motor domain targets Dyn1/HC to microtubule plus ends in a Bik1/CLIP-170- and Pac1/LIS1-dependent manner. Surprisingly, the isolated motor domain blocks plus-end targeting of Dyn1/HC, leading to a dominant-negative effect on dynein function. Overexpression of Pac1/LIS1, but not Bik1/CLIP-170, rescues the dominant negativity by restoring Dyn1/HC to plus ends. In contrast, the isolated tail domain has no inhibitory effect on Dyn1/HC targeting and function. However, cortical targeting of the tail construct is more robust than full-length Dyn1/HC and occurs independently of Bik1/CLIP-170 or Pac1/LIS1. CONCLUSIONS: Our results suggest that the cortical association domain is normally masked in the full-length dynein molecule. We propose that targeting of dynein to plus ends unmasks the tail, priming the motor for off-loading to cortical Num1 sites. PMID- 19185496 TI - The coding of color, motion, and their conjunction in the human visual cortex. AB - BACKGROUND: Color and motion serve as the prime examples of segregated processing in the visual brain, giving rise to the question how color-motion conjunctions are represented. This problem is also known as the "binding problem." RESULTS: Human volunteers viewed visual displays containing colored dots rotating around the center. The dots could be red or green and rotate clockwise or counterclockwise, leading to four possible stimulus displays. Superimposed pairs of such stimuli provided two additional displays, each containing both colors and both directions of motion but differing in their feature conjunctions. We applied multivariate classifiers to voxel-activation patterns obtained while subjects viewed such displays. Our analyses confirm the presence of directional-motion information across visual cortex and provide evidence of hue coding in all early visual areas except V5/MT(+). Within each cortical area, information on color and motion appeared to be coded in distinct sets of voxels. Furthermore, our results demonstrate the explicit representation of feature conjunctions in the primary visual cortex and beyond. CONCLUSIONS: The results show that conjunctions can be decoded from spatial activation patterns already in V1, indicating an explicit coding of conjunctions at early stages of visual processing. Our findings raise the possibility that the solution of what has been taken as the prime example of the binding problem engages neural mechanisms as early as V1. PMID- 19185497 TI - Social feedback and the emergence of leaders and followers. AB - In many animal groups, certain individuals consistently appear at the forefront of coordinated movements [1-4]. How such leaders emerge is poorly understood [5, 6]. Here, we show that in pairs of sticklebacks, Gasterosteus aculeatus, leadership arises from individual differences in the way that fish respond to their partner's movements. Having first established that individuals differed in their propensity to leave cover in order to look for food, we randomly paired fish of varying boldness, and we used a Markov Chain model to infer the individual rules underlying their joint behavior. Both fish in a pair responded to each other's movements-each was more likely to leave cover if the other was already out and to return if the other had already returned. However, we found that bolder individuals displayed greater initiative and were less responsive to their partners, whereas shyer individuals displayed less initiative but followed their partners more faithfully; they also, as followers, elicited greater leadership tendencies in their bold partners. We conclude that leadership in this case is reinforced by positive social feedback. PMID- 19185498 TI - phytochrome B and PIF4 regulate stomatal development in response to light quantity. AB - Stomata are pores on the surfaces of leaves that regulate gas exchange between the plant interior and the atmosphere [1]. Plants adapt to changing environmental conditions in the short term by adjusting the aperture of the stomatal pores, whereas longer-term changes are accomplished by altering the proportion of stomata that develop on the leaf surface [2, 3]. Although recent work has identified genes involved in the control of stomatal development [4], we know very little about how stomatal development is modulated by environmental signals, such as light. Here, we show that mature leaves of Arabidopsis grown at higher photon irradiances show significant increases in stomatal index (S.I.) [5] compared to those grown at lower photon irradiances. Light quantity-mediated changes in S.I. occur in red light, suggesting that phytochrome photoreceptors [6] are involved. By using a genetic approach, we demonstrate that this response is dominated by phytochrome B and also identify a role for the transcription factor, PHYTOCHROME-INTERACTING FACTOR 4 (PIF4) [7]. In sum, we identify a photoreceptor and downstream signaling protein involved in light-mediated control of stomatal development, thereby establishing a tractable system for investigating how an environmental signal modulates stomatal development. PMID- 19185499 TI - Clinical evaluation of the proximal tibiofibular joint in knees with severe tibiofemoral primary osteoarthritis. AB - Despite the coexistence of primary arthritic involvement of the tibiofemoral and proximal tibiofibular joints (PTFJ), clinical features of the PTFJ in knees with severe femorotibial arthritis have not been described to date. The PTFJ in knees with primary osteoarthritis may be a source of lateral knee pain after total knee arthroplasty operations. This study seeks to address whether there is a relationship between the clinical findings and degeneration of the PTFJ in knees with severe tibiofemoral primary osteoarthritis. Sixty knees in 34 consecutive patients (31 females and 3 males; mean age 71 years; range 61-86 years) with Kellgren-Lawrence grades III-IV primary femorotibial arthritis were enrolled in this study in order to collect clinical data regarding the PTFJ. Radiographs showed 23 PTFJs were grade IV, 14 were grade III, and 23 were grade II. With regard to the type of PTFJ, 13 joints were horizontal and 47 were oblique. Pain and tenderness were observed in 13 PTFJs upon clinical exam (two horizontal and 11 oblique PTFJ types; grade II in five, grade III in two, and grade IV in six). Lateral hamstring tightness was found in 28 knees (24 oblique and four horizontal PTFJ types; grade II in 10, grade III in two, and grade IV in 16). Ten of the 28 PTFJs in knees with lateral hamstring tightness were painful during the physical examination. No significant relationship was found between PTFJ exam findings and PTFJ type (Fisher's Exact test, p=0.713), PTFJ grade of arthritis (chi(2) test, p=0.700), or between hamstring tightness and PTFJ type (chi(2) test, p=0.194). However, hamstring tightness was significantly correlated with the grade of arthritis in the PTFJ (chi(2) test, p=0.004). Although degenerative disease of the PTFJ is commonly associated with degenerative disease of the knee joint, radiographic findings of the PTFJ in patients with severe degenerative knee osteoarthritis and varus malalignment do not correlate with clinical findings. PMID- 19185500 TI - Clinical, radiological and histological evaluation of biphasic calcium phosphate bioceramic wedges filling medial high tibial valgisation osteotomies. AB - We report clinical, radiological and histological findings following high tibial valgisation osteotomy (HTVO) using micro-macroporous biphasic calcium phosphate wedges fixed with a plate and locking screws. From 1999 to 2002, 43 knees were operated on and studied prospectively. All underwent clinical and radiological follow-up at days 1, 90, and 365 to evaluate consolidation and bone substitute interfaces. Additionally, biopsies were taken for histology at least 1 year after implantation from 10 patients who requested plate removal. Radiologically, consolidation was observed in 98% of cases. At 1 year, correction was unchanged in 95% of cases. Histological analysis revealed considerable MBCP resorption and bone ingrowth, both into the pores and replacing the bioceramic material. Polarised light microscopy confirmed normal bony architecture with trabecular and/or dense lamellar bone growth at the expense of the wedge implants. X-ray and micro-CT scan revealed a well organised and mineralised structure in the newly formed bone. This study shows that using MBCP wedges in combination with orientable locking screws and a plate is a simple, safe and fast surgical technique for HTVO. The is the first study to examine the results by histological analysis, which confirmed good outcomes. PMID- 19185501 TI - Novel E. coli beta-ketoacyl-acyl carrier protein synthase III inhibitors as targeted antibiotics. AB - Beta-ketoacyl-acyl carrier protein synthase (KAS) III is a condensing enzyme that initiates fatty acid biosynthesis in most bacteria. We determined three pharmacophore maps from receptor-oriented pharmacophore-based in silico screening of the X-ray structure of Escherichia coli KAS III (ecKAS III) and choose 16 compounds as candidate ecKAS III inhibitors. Binding inhibitors were characterized using saturation-transfer difference NMR spectroscopy (STD-NMR), and binding constants were determined with fluorescence quenching experiments. Based on the results, we propose that the antimicrobial compound, 4 cyclohexyliminomethyl-benzene-1,3-diol (YKAs3003), is a potent inhibitor of pathogenic KAS III, displaying minimal inhibitory concentration (MIC) values in the range 128-256 microg/mL against various bacteria. PMID- 19185503 TI - A new generation of biocides for control of crustacea in fish farms. AB - Farming of salmon has become a significant industry in many countries over the past two decades. A major challenge facing this sector is infestation of the salmon by sea lice. The main way of treating salmon for such infestations is the use of medicines such as organophosphates, pyrethrins, hydrogen peroxide or benzoylphenyl ureas. The use of these medicines in fish farms is, however, highly regulated due to concerns about contamination of the wider marine environment. In this paper we report the use of photochemically active biocides for the treatment of a marine copepod, which is a model of parasitic sea lice. Photochemical activation and subsequent photodegradation of PDAs may represent a controllable and environmentally benign option for control of these parasites or other pest organisms in aquaculture. PMID- 19185505 TI - Role of RANKL in bone diseases. AB - Bone remodeling is a tightly regulated process of osteoclast-mediated bone resorption, balanced by osteoblast-mediated bone formation. Disruption of this balance can lead to increased bone turnover, resulting in excessive bone loss or extra bone formation and consequent skeletal disease. The receptor activator of nuclear factor kappaB ligand (RANKL) (along with its receptor), the receptor activator of nuclear factor kappaB and its natural decoy receptor, osteoprotegerin, are the final effector proteins of osteoclastic bone resorption. Here, I provide an overview of recent studies that highlight the key role of RANKL in the pathophysiology of several bone diseases and discuss the novel therapeutic approaches afforded by the modulation of RANKL. PMID- 19185504 TI - Cellular fatty acid uptake: a pathway under construction. AB - Membrane uptake of long-chain fatty acids (FAs) is the first step in cellular FA utilization and a point of metabolic regulation. CD36 facilitates a major fraction of FA uptake by key tissues. This review highlights the contribution of CD36 to pathophysiology in rodents and humans. Novel concepts regarding regulation of CD36-facilitated uptake are discussed (i.e. the role of membrane rafts and caveolae, CD36 recycling between intracellular depots and the membrane, and chemical modifications of the protein that impact its turnover and recruitment). Importantly, CD36 membrane levels and turnover are abnormal in diabetes, resulting in dysfunctional FA utilization. In addition, variants in the CD36 gene were shown recently to influence susceptibility for the metabolic syndrome, which greatly increases the risk of diabetes and heart disease. PMID- 19185506 TI - Performance of the whole-blood stimulation assay for assessing innate immune activation under field conditions. AB - Innate propensity of immune activation is reflected in production of pro- and anti-inflammatory cytokines upon stimulation of Toll-like receptors (TLR) in whole-blood stimulation assays. The validity of the whole-blood stimulation assay under field conditions has not been evaluated extensively. Here, we have determined correlation of individually repeated whole-blood stimulation assays in a field-study in Ghana and compared it with that of two Dutch populations performed under optimal conditions. We also examined cytokine production to various TLR-agonists in order to create an assay that would mimic general innate immune responses. Under field conditions repeated assessments of lipopolysaccharide-induced Tumor Necrosis Factor-alpha (TNFalpha) production were poorly correlated (r=0.15, p=0.087). Correlation was relatively high for production of Interleukin-10 (IL10) (r=0.48, p<0.001) and comparable to that observed in the Dutch population under optimal conditions. Combined stimulation with lipopolysaccharide and zymosan resulted in cytokine production profiles that were similar to that attained after stimulation with a mixed culture of bacteria. Here, we conclude that variation of a whole-blood assay performed in field setting is large in general but that production of IL10 seems to better reflect an innate pro- or anti-inflammatory tendency whereas production of TNFalpha may predominantly reflect recent immunological challenges. Furthermore, simultaneous stimulation of several Toll-like receptors may mimic general innate immune activation. PMID- 19185507 TI - Common polymorphisms in interleukin genes (IL4, IL6, IL8 and IL12) are not associated with alcoholic liver disease or alcoholism in Spanish men. AB - BACKGROUND: Preliminary data suggest that polymorphisms in cytokine genes may be involved in the genetic predisposition to alcoholic liver cirrhosis or alcohol use disorders. We thus analyze the association between these diseases and the following polymorphisms: -33T>C IL4, -174 G>C IL6, -251 T>A IL8 and 1188 A>C IL12B. METHODS: 258 male alcoholics (161 without liver disease and 97 with liver cirrhosis) and 101 healthy controls were genotyped for the above mentioned polymorphisms. We examined the relationship between genotype and allele frequencies and the presence of disease, as well as the correlation with combinations of putative pro-inflammatory genotypes. Haplotypes were inferred using the expectation-maximization algorithm and haplotype frequencies were compared. RESULTS: We found no statistically significant association between any of these polymorphisms or the combinations of pro-inflammatory polymorphisms and the risk of alcoholic liver cirrhosis or alcohol abuse or dependence. Haplotype analysis of the IL4 and IL12B polymorphisms did not show any statistical relationship either. CONCLUSIONS: Our results do not support the hypothesis that the analyzed polymorphisms confer differences in alcoholic liver cirrhosis or alcohol use disorders susceptibility. PMID- 19185508 TI - Ab initio investigation of the hydration of deprotonated amino acids. AB - The complexation of five deprotonated anionic amino acids (glycine, L-alanine, L valine, L-Aspartic acid, and L-glutamine) with one water molecule, has been investigated using a MP2/63-11++G(d,p) approach fully accounting for the basis set superposition errors. For each amino acid, several energetic minima have been identified, and we provide spectroscopic information allowing to discriminate them. Our results strongly suggest that two complexes should coexist under the experimental conditions for [Ala - H](-), [Val - H](-), and [Asp - H](-). Comparisons with the experimental enthalpies, entropies, and Gibbs free energies recently obtained by Wincel [J. Am. Soc. Mass Spectrom. 2008, 19, 1091-1097] show that our simulation reproduces the most significant structure/energy experimental trends, though the entropic changes induced by hydration are slightly overestimated. PMID- 19185509 TI - Host-guest hydrogen atom transfer induced by electron capture. AB - 1,n-Alkanediammonium cations in noncovalent complexes with two dibenzo-18-crown-6 ether (DBCE) ligands undergo an unusual intramolecular tandem hydrogen atom and proton transfer to the crown ether ligand upon charge reduction by electron capture. Deuterium labeling established that both migrating hydrogens originated from the ammonium groups. The double hydrogen transfer was found to depend on the length of the alkane chain connecting the ammonium groups. Ab initio calculations provided structures for select alkanediammonium.dibenzo-18-crown-6-ether complexes and dissociation products. This first observation of an intra-complex hydrogen transfer is explained by the unusual electronic properties of the complexes and the substantial hydrogen atom affinity of the aromatic rings in the crown ligand. PMID- 19185511 TI - Gas-phase reactivity of ruthenium carbonyl cluster anions. AB - Partially-ligated anionic ruthenium carbonyl clusters react with alkenes, arenes, and alkanes in the gas phase; the products undergo extensive C-H activation and lose dihydrogen and carbon monoxide under collision-induced dissociation conditions. Triethylsilane and phenylsilane are also reactive towards the unsaturated clusters, and oxygen was shown to rapidly break down the cluster core by oxidative cleavage of the metal-metal bonds. These qualitative gas-phase reactivity studies were conducted using an easily-installed and inexpensive modification of a commercial electrospray ionization mass spectrometer. Interpretation of the large amounts of data generated in these studies is made relatively straightforward by employing energy-dependent electrospray ionization mass spectrometry (EDESI-MS). PMID- 19185510 TI - Deciphering the peptide iodination code: influence on subsequent gas-phase radical generation with photodissociation ESI-MS. AB - Iodination of tyrosine was recently discovered as a useful method for generating radical peptides via photodissociation of carbon-iodine bonds by an ultraviolet photon in the gas phase. The subsequent fragmentation behavior of the resulting odd-electron peptides is largely controlled by the radical. Although previous experiments have focused on mono-iodination of tyrosine, peptides and proteins can also be multiply iodinated. Tyrosine and, to a lesser extent, histidine can both be iodinated or doubly iodinated. The behavior of doubly iodinated residues is explored under conditions where the sites of iodination are carefully controlled. It is found that radical peptides generated by the loss of a single iodine from doubly iodinated tyrosine behave effectively identically to singly iodinated peptides. This suggests that the remaining iodine does not interfere with radical directed dissociation pathways. In contrast, the concerted loss of two iodines from doubly iodinated peptides yields substantially different results that suggest that radical recombination can occur. However, sequential activation can be used to generate multiple usable radicals in different steps of an MS(n) experiment. Furthermore, it is demonstrated that in actual peptides, the rate of iodination for tyrosine versus mono-iodotyrosine cannot be predicted easily a priori. In other words, previous assumptions that mono-iodination of tyrosine is the rate-limiting step to the formation of doubly iodinated tyrosine are incorrect. PMID- 19185512 TI - Intact and top-down characterization of biomolecules and direct analysis using infrared matrix-assisted laser desorption electrospray ionization coupled to FT ICR mass spectrometry. AB - We report the implementation of an infrared laser onto our previously reported matrix-assisted laser desorption electrospray ionization (MALDESI) source with ESI post-ionization yielding multiply charged peptides and proteins. Infrared (IR)-MALDESI is demonstrated for atmospheric pressure desorption and ionization of biological molecules ranging in molecular weight from 1.2 to 17 kDa. High resolving power, high mass accuracy single-acquisition Fourier transform ion cyclotron resonance (FT-ICR) mass spectra were generated from liquid- and solid state peptide and protein samples by desorption with an infrared laser (2.94 mum) followed by ESI post-ionization. Intact and top-down analysis of equine myoglobin (17 kDa) desorbed from the solid state with ESI post-ionization demonstrates the sequencing capabilities using IR-MALDESI coupled to FT-ICR mass spectrometry. Carbohydrates and lipids were detected through direct analysis of milk and egg yolk using both UV- and IR-MALDESI with minimal sample preparation. Three of the four classes of biological macromolecules (proteins, carbohydrates, and lipids) have been ionized and detected using MALDESI with minimal sample preparation. Sequencing of O-linked glycans, cleaved from mucin using reductive beta elimination chemistry, is also demonstrated. PMID- 19185513 TI - Measurement of neuropeptides in crustacean hemolymph via MALDI mass spectrometry. AB - Neuropeptides are often released into circulatory fluid (hemolymph) to act as circulating hormones and regulate many physiological processes. However, the detection of these low-level peptide hormones in circulation is often complicated by high salt interference and rapid degradation of proteins and peptides in crude hemolymph extracts. In this study, we systematically evaluated three different neuropeptide extraction protocols and developed a simple and effective hemolymph preparation method suitable for MALDI MS profiling of neuropeptides by combining acid-induced abundant protein precipitation/depletion, ultrafiltration, and C(18) micro-column desalting. In hemolymph samples collected from the crab Cancer borealis, several secreted neuropeptides have been detected, including members from at least five neuropeptide families, such as RFamide, allatostatin, orcokinin, tachykinin-related peptide (TRP), and crustacean cardioactive peptide (CCAP). Furthermore, two TRPs were detected in the hemolymph collected from food deprived animals, suggesting the potential role of these neuropeptides in feeding regulation. In addition, a novel peptide with a Lys-Phe-amide C-terminus was identified and de novo sequenced directly from the Cancer borealis hemolymph sample. To better characterize the hemolymph peptidome, we also identified several abundant peptide signals in C. borealis hemolymph that were assigned to protein degradation products. Collectively, our study describes a simple and effective sample preparation method for neuropeptide analysis directly from crude crustacean hemolymph. Numerous endogenous neuropeptides were detected, including both known ones and new peptides whose functions remain to be characterized. PMID- 19185514 TI - Differentiation of 3-O-sulfated heparin disaccharide isomers: identification of structural aspects of the heparin CCL2 binding motif. AB - The presence of 3-O-sulfated glucosamine residues in heparin or heparan sulfate plays a role in binding to antithrombin III and HSV infection. In this study, tandem mass spectrometry was used to differentiate between two heparin disaccharide isomers containing variable sulfate at C6 in a common disaccharide and C3 in a more rare one. The dissociation patterns shown by MS(2) and MS(3) were clearly distinguishable between the isomers, allowing their differentiation and quantitation. Using this technique, we show that an octasaccharide with 11 sulfate groups with high affinity for inflammatory chemokine CCL2 does not contain 3-O-sulfated disaccharides. PMID- 19185515 TI - Characterization of direct-current atmospheric-pressure discharges useful for ambient desorption/ionization mass spectrometry. AB - Two relatively new ambient ionization sources, direct analysis in real time (DART) and the flowing atmospheric-pressure afterglow (FAPA), use direct current, atmospheric-pressure discharges to produce reagent ions for the direct ionization of a sample. Although at a first glance these two sources appear similar, a fundamental study reveals otherwise. Specifically, DART was found to operate with a corona-to-glow transition (C-G) discharge whereas the FAPA was found to operate with a glow-to-arc transition (G-A) discharge. The characteristics of both discharges were evaluated on the basis of four factors: reagent-ion production, response to a model analyte (ferrocene), infrared (IR) thermography of the gas used for desorption and ionization, and spatial emission characteristics. The G-A discharge produced a greater abundance and a wider variety of reagent ions than the C-G discharge. In addition, the discharges yielded different adducts and signal strengths for ferrocene. It was also found that the gas exiting the discharge chamber reached a maximum of 235 degrees C and 55 degrees C for the G-A and C-G discharges, respectively. Finally, spatially resolved emission maps of both discharges showed clear differences for N(2)(+) and O(I). These findings demonstrate that the discharges used by FAPA and DART are fundamentally different and should have different optimal applications for ambient desorption/ionization mass spectrometry (ADI-MS). PMID- 19185516 TI - Invisible motion contributes to simultaneous motion contrast. AB - The purpose of the present study was two-fold. First we examined whether visible motion appearance was altered by the spatial interaction between invisible and visible motion. We addressed this issue by means of simultaneous motion contrast, in which a horizontal test grating with a counterphase luminance modulation was seen to have the opposite motion direction to a peripheral inducer grating with unidirectional upward or downward motion. Using a mirror stereoscope, observers viewed the inducer and test gratings with one eye, and continuous flashes of colorful squares forming an annulus shape with the other eye. The continuous flashes rendered the inducer subjectively invisible. The observers' task was to report whether the test grating moved upward or downward. Consequently, simultaneous motion contrast was observed even when the inducer was invisible (Experiment 1). Second, we examined whether the observers could correctly respond to the direction of invisible motion: It was impossible (Experiment 2). PMID- 19185517 TI - Conduits mediate transport of low-molecular-weight antigen to lymph node follicles. AB - To track drainage of lymph-borne small and large antigens (Ags) into the peripheral lymph nodes and subsequent encounter by B cells and follicular dendritic cells, we used the approach of multiphoton intravital microscopy. We find a system of conduits that extend into the follicles and mediate delivery of small antigens to cognate B cells and follicular dendritic cells. The follicular conduits provide an efficient and rapid mechanism for delivery of small antigens and chemokines such as CXCL13 to B cells that directly contact the conduits. By contrast, large antigens were bound by subcapsular sinus macrophages and subsequently transferred to follicular B cells as previously reported. In summary, the findings identify a unique pathway for the channeling of small lymph borne antigens and chemoattractants from the subcapsular sinus directly to the B cell follicles. This pathway could be used for enhancing delivery of vaccines or small molecules for improvement of humoral immunity. PMID- 19185519 TI - Risk factors and consequences of early childhood dyssomnias: New perspectives. AB - Dyssomnias are largely under-diagnosed in infants and toddlers. This literature review proposes an integrative model based on empirical data on determinants and consequences of sleep disturbances occurring in early life. This model proposes that parental behaviors that impede the child's autonomy toward sleep periods are primary grounds for the development of dyssomnias, e.g., parental presence until the child falls asleep, and putting an already sleeping child to bed. The model also indicates the serious potential consequences of a modest but chronic loss of sleep in childhood. At least three developmental domains could be directly affected: behavioral/social competence, cognitive performance, and physical condition. Thus, children with short nocturnal sleep duration before age 3.5 years show increased risk of high hyperactivity-impulsivity scores and low cognitive performance at 6 years compared to children who sleep 11 h per night, after controlling for potentially confounding variables. Moreover, persistent short sleep duration in early infancy increased the risk of suffering of obesity at 6 years of age, after controlling for potentially confounding variables. Finally, the importance of allowing the child to sleep at least 10 h per night in early childhood is stressed, as the National Sleep Foundation Poll suggests, for optimal child development. PMID- 19185518 TI - A low interleukin-2 receptor signaling threshold supports the development and homeostasis of T regulatory cells. AB - Interleukin-2 receptor (IL-2R) signaling is essential for T regulatory (Treg) cell development and homeostasis. Here, we show that expression of IL-2Rbeta chains that lack tyrosine residues important for the association of the adaptor Shc and the transcription factor STAT5 in IL-2Rbeta-deficient mice resulted in production of a normal proportion of natural Treg cells that suppressed severe autoimmunity related with deficiency in IL-2 or IL-2R. These mutant IL-2Rbeta chains supported suboptimal and transient STAT5 activation that upregulate the transcription factor Foxp3 to normal amounts in natural, but not induced, Treg cells. Nevertheless, gene expression profiling revealed many targets in peripheral natural Treg cells that were IL-2 dependent and a substantial overlap between the Treg cell IL-2-dependent gene program and the Treg cell transcriptional signature. Collectively, these findings demonstrate that a critical, and perhaps minor, subset of IL-2-dependent targets is indexed to a low IL-2R signaling threshold and that a substantial proportion of the Treg cell gene program is regulated by IL-2. PMID- 19185520 TI - Influence of a cyclic combination chemotherapeutic protocol on primary haemostasis in dogs suffering from malignant lymphoma. AB - The purpose of this study was to examine the influence of cyclic combination chemotherapy on primary haemostasis in dogs with malignant lymphoma. Seventeen dogs receiving cytostatic treatment for high-grade lymphoma were included in the study. The dogs were treated with a Madison-Wisconsin derived protocol, which included asparaginase, vincristine, doxorubicin and prednisolone. At different time points during the first 4 weeks of induction, platelet count, capillary bleeding time, analysis of the platelet function using the platelet function analyser PFA-100, and platelet aggregation by the Born-method were measured. The most obvious changes were found for median values of the platelet count, which increased significantly from 210,000/microL before induction to 349,000/microL during the second week of induction (P=0.0010). Median platelet count subsequently decreased by the fourth week of treatment (Friedman-test: P<0.0001). None of the parameters of platelet function (capillary bleeding time, automatic platelet function analysis, aggregation maximum) showed significant changes with time (P>0.05, Friedman-test). The results did not suggest that significant platelet dysfunction was induced by the chemotherapeutic protocol used in the study. PMID- 19185521 TI - Robust and efficient spin-locked symmetry-based double-quantum homonuclear dipolar recoupling for probing (1)H-(1)H proximity in the solid-state. AB - We report a novel symmetry-based method, using inversion elements bracketed by spin locks, for exciting double-quantum (DQ) coherences between spin-1/2 nuclei, such as protons. Compared to previous DQ-recoupling techniques, this new pulse sequence requires moderate rf field, even at ultra-fast MAS speeds. Furthermore, it is easy to implement and it displays higher robustness to both chemical shift anisotropy and to spreads in resonance frequencies. These advances greatly facilitate the observation of (1)H-(1)H proximities at high fields and high MAS frequencies. PMID- 19185522 TI - Narrow peaks and high dimensionalities: exploiting the advantages of random sampling. AB - Level of artifacts in spectra obtained by Multidimensional Fourier Transform has been studied, considering randomly sampled signals of high dimensionality and long evolution times. It has been shown theoretically and experimentally, that this level is dependent on the number of time domain samples, but not on its relation to the number of points required in appropriate conventional experiment. Independence of the evolution time domain size (in the terms of both: dimensionality and evolution time reached), suggests that random sampling should be used rather to design new techniques with large time domain than to accelerate standard experiments. 5D HC(CC-TOCSY)CONH has been presented as the example of such approach. The feature of Multidimensional Fourier Transform, namely the possibility of calculating spectral values at arbitrary chosen frequency points, allowed easy examination of resulting spectrum. We present the example of such approach, referred to as Sparse Multidimensional Fourier Transform. PMID- 19185524 TI - Structural basis for recognition of diubiquitins by NEMO. AB - NEMO is the regulatory subunit of the IkappaB kinase (IKK) in NF-kappaB activation, and its CC2-LZ region interacts with Lys63 (K63)-linked polyubiquitin to recruit IKK to receptor signaling complexes. In vitro, CC2-LZ also interacts with tandem diubiquitin. Here we report the crystal structure of CC2-LZ with two dimeric coiled coils representing CC2 and LZ, respectively. Surprisingly, mutagenesis and nuclear magnetic resonance experiments reveal that the binding sites for diubiquitins at LZ are composites of both chains and that each ubiquitin in diubiquitins interacts with symmetrical NEMO asymmetrically. For tandem diubiquitin, the first ubiquitin uses the conserved hydrophobic patch and the C-terminal tail, while the second ubiquitin uses an adjacent surface patch. For K63-linked diubiquitin, the proximal ubiquitin uses its conserved hydrophobic patch, while the distal ubiquitin mostly employs the C-terminal arm including the K63 linkage residue. These studies uncover the energetics and geometry for mutual recognition of NEMO and diubiquitins. PMID- 19185525 TI - Tuberculosis elimination in the Canadian First Nations population: assessment by a state-transfer, compartmental epidemic model. AB - BACKGROUND: Tuberculosis (TB) remains an important public health problem in Canadian Aboriginal (First Nations and Inuit) communities. The objectives of this study were to predict future disease burden and set feasible targets for the elimination of TB in the First Nations population, using retrospective data and an epidemic model. METHODS: Reported TB incidence data (1974-2002), previously published TB meningitis data from the pre-chemotherapy era, and previous estimates of disease risk following infection were used to estimate a trend in the annual risk of infection from 1929 to 2002, and the age-specific prevalence of infection in 2002. A state-transfer, compartmental model was then developed to predict future disease burden. Two scenarios were simulated, with different disease risk parameters. RESULTS: The estimated prevalence of infection in 2002 was 20.9% in scenario 1 and 25.5% in scenario 2. Predicted incidence rates in 2015 were 16.8 per 100,000 and 11.7 per 100,000 for the two scenarios, respectively. The incidence of disease was not lower than 1 per 100,000 for either scenario in 2034, the arbitrarily chosen last year of the model. CONCLUSIONS: The goal of eliminating TB among Aboriginal peoples in Canada is a feasible one, but will only be achieved with continued investment in programs designed to control and prevent transmission. Reactivation disease cases may occur for a number of years to come, making rapid elimination a difficult goal. PMID- 19185523 TI - A novel NDUFA1 mutation leads to a progressive mitochondrial complex I-specific neurodegenerative disease. AB - Mitochondrial diseases have been shown to result from mutations in mitochondrial genes located in either the nuclear DNA (nDNA) or mitochondrial DNA (mtDNA). Mitochondrial OXPHOS complex I has 45 subunits encoded by 38 nuclear and 7 mitochondrial genes. Two male patients in a putative X-linked pedigree exhibiting a progressive neurodegenerative disorder and a severe muscle complex I enzyme defect were analyzed for mutations in the 38 nDNA and seven mtDNA encoded complex I subunits. The nDNA X-linked NDUFA1 gene (MWFE polypeptide) was discovered to harbor a novel missense mutation which changed a highly conserved glycine at position 32 to an arginine, shown to segregate with the disease. When this mutation was introduced into a NDUFA1 null hamster cell line, a substantial decrease in the complex I assembly and activity was observed. When the mtDNA of the patient was analyzed, potentially relevant missense mutations were observed in the complex I genes. Transmitochondrial cybrids containing the patient's mtDNA resulted in a mild complex I deficiency. Interestingly enough, the nDNA encoded MWFE polypeptide has been shown to interact with various mtDNA encoded complex I subunits. Therefore, we hypothesize that the novel G32R mutation in NDUFA1 is causing complex I deficiency either by itself or in synergy with additional mtDNA variants. PMID- 19185526 TI - Pott's disease with bilateral paraspinal abscesses. PMID- 19185527 TI - Impact of sampling rate for time domain analysis of continuous intracranial pressure (ICP) signals. AB - Time domain analysis of the intracranial pressure (ICP) waveform is critically dependent on the exact reproduction of the ICP waveform. This study explored how the sampling rate of the ICP signal affects the time domain analysis. It was also assessed through this study how upsampling (interpolation) improves the time domain analysis. From the hospital database, a set of 55 ICP waveforms were retrieved from 48 patients (28 children and 20 adults). First, the ICP signals originally sampled at 200 or 100 Hz were compared with the ICP signals downsampled to 5, 10, 20, 25, 50 Hz (and 100 Hz). Second, the original ICP signals were compared with ICP signals upsampled (i.e. interpolated) to 100 Hz (from 5, 10, 20, 25 or 50 Hz). For each ICP recording the output of time domain analysis was the average value and the quantitative distribution of mean ICP wave amplitudes determined every six second (6 s) time window. The total material incorporated a total of 373,371 6 s time windows. Downsampling revealed that the time domain analysis could be most faithfully applied to ICP signals sampled at 50 Hz or above, while ICP signals sampled at 25 Hz deviated more from the original signal than we would accept from a clinical perspective. The use of interpolation gave better representation of the peaks, and should be applied to all ICP signals sampled at lower rate than 100 Hz. PMID- 19185528 TI - Prescribing soft contact lenses for astigmatism. AB - There have been continued improvements in toric soft lens design over the past three decades. Data that we have gathered from annual contact lens fitting surveys demonstrate a commensurate increase in toric lens fitting as a proportion of all soft lenses fitted. The current toric lens prescribing rate (34% of all lenses when cosmetic tints, monovision and multifocal lenses are ignored) is roughly equivalent to the rate that would be achieved if all cases of astigmatism 1.00D or more were fitted with toric lenses. Toric lenses tend to be fitted more to males and prescribed for monthly rather than daily replacement. PMID- 19185529 TI - DNA changes tell us about rice domestication. AB - Crop domestication can be considered a model system of plant evolution. Genome analyses of rice have revealed the fine population structure of this major crop associated with local origins of landraces. Recent cloning of rice domestication related genes and identification of the responsible functional nucleotide polymorphisms in landraces, while taking into account their population structures, have revealed the existence of historical signatures of the DNA involved in the domestication process. These signatures imply the importance of multiple selection steps wherein natural variants were combined to improve crop performance during domestication. These analyses will provide new insights into the relationship between Darwinian selection for agronomical phenotypes and DNA changes in terms of plant evolution. PMID- 19185530 TI - The emergence of whole genome association scans in barley. AB - Barley geneticists are currently using association genetics to identify and fine map traits directly in elite plant breeding material. This has been made possible by the development of a highly parallel SNP assay platform that provides sufficient marker density for genome-wide scans and linkage disequilibrium-led gene identification. By leveraging the combined resources of the barley research and breeding sectors, marker-trait associations are being identified and a renewed interest has emerged in novel strategies for barley improvement. New database and visualization tools have been developed and statistical methods adapted from human genetics to account for complexities in the datasets. Exciting early results suggest that association genetics will assume a central role in establishing genotype-to-phenotype relationships. PMID- 19185531 TI - Translocated effectors of Yersinia. AB - Currently, all known translocated effectors of Yersinia are delivered into host cells by type III secretion systems (T3SSs). Pathogenic Yersinia maintain the plasmid-encoded Ysc T3SS for the specific delivery of the well-studied Yop effectors. New horizons for effector biology have opened with the discovery of the Ysps of Y. enterocolitica Biovar 1B, which are translocated into host cells by the chromosome-endoded Ysa T3SS. The reported arsenal of effectors is likely to expand since genomic analysis has revealed gene-clusters in some Yersinia that code for other T3SSs. These efforts also revealed possible type VI secretion (T6S) systems, which may indicate that translocation of effectors occurs by multiple mechanisms. PMID- 19185532 TI - An evaluation of least-squares fitting methods in XAFS spectroscopy: iron-based SBA-15 catalyst formulations. AB - A detailed comparison has been made of determinations by (57)Fe Mossbauer spectroscopy and four different XAFS spectroscopic methods of %Fe as hematite and ferrihydrite in 11 iron-based SBA-15 catalyst formulations. The four XAFS methods consisted of least-squares fitting of iron XANES, d(XANES)/dE, and EXAFS (k(3)chi and k(2)chi) spectra to the corresponding standard spectra of hematite and ferrihydrite. The comparison showed that, for this particular application, the EXAFS methods were superior to the XANES methods in reproducing the results of the benchmark Mossbauer method in large part because the EXAFS spectra of the two iron-oxide standards were much less correlated than the corresponding XANES spectra. Furthermore, the EXAFS and Mossbauer results could be made completely consistent by inclusion of a factor of 1.3+/-0.05 for the ratio of the Mossbauer recoilless fraction of hematite relative to that of ferrihydrite at room temperature (293K). This difference in recoilless fraction is attributed to the nanoparticle nature of the ferrihydrite compared to the bulk nature of the hematite. Also discussed are possible alternative non-least-squares XAFS methods for determining the iron speciation in this application as well as criteria for deciding whether or not least-squares XANES methods should be applied for the determination of element speciation in unknown materials. PMID- 19185533 TI - Synthesis, characterization, biological activities of dimethyltin(IV) complexes of Schiff bases with ONO-type donors. AB - Four different dimethyltin(IV) complexes of Schiff bases derived from 2-amino-3 hydroxypyridine and different substituted salicylaldehydes have been synthesized. The compounds, with the general formula [Me(2)Sn(2-OArCH=NC(5)H(3)NO)], where Ar= C(6)H(3)(5-CH(3)) [Me(2)SnL(1)], -C(6)H(3)(5-NO(2)) [Me(2)SnL(2)], -C(6)H(2)(3,5 Cl(2)) [Me(2)SnL(3)], and -C(6)H(2)(3,5-I(2)) [Me(2)SnL(4)], were characterized by IR, NMR ((1)H and (13)C), mass spectroscopy and elemental analysis. Me(2)SnL(3) was also characterized by X-ray diffraction analysis and shows a fivefold C(2)NO(2) coordination with distorted square pyramidal geometry. H(3)C Sn-CH(3) angles in the complexes were calculated using Lockhart's equations with the (1)J((117/119)Sn-(13)C) and (2)J((117/119)Sn-(1)H) values (from the (1)H-NMR and (13)C-NMR spectra). The in vitro antibacterial and antifungal activities of dimethyltin(IV) complexes were also investigated. PMID- 19185534 TI - Density functional theory study on surface-enhanced Raman scattering of 4,4' azopyridine on silver. AB - Surface-enhanced Raman scattering (SERS) of 4,4'-azopyridine (AZPY) on silver foil substrate was measured under 1064nm excitation lines. Density-functional theory (DFT) methods were used to calculate the structure and vibrational spectra of models such as Ag-AZPY, Ag(4)-AZPY and Ag(6)-AZPY complexes with B3LYP/6 31++G(d,p)(C,H,N)/Lanl2dz(Ag) basis set. The Raman bands of AZPY were identified on the ground of analog computation of potential energy distribution. The calculated spectra of Ag(4)-AZPY and Ag(6)-AZPY models were much approximated to the experimental results than that of Ag-AZPY model. The DFT results showed that the angles between two pyridyl rings keep 0 degrees from AZPY to Ag-AZPY, Ag(4) AZPY and Ag(6)-AZPY model. The energy gaps between the HOMO and LUMO changed from 363 to 1140nm for AZPY-Ag complexes according to the DFT results. An conclusion was conceived that chemical enhancement mechanism may play an important role in the SERS of AZPY on silver substrate. PMID- 19185535 TI - Probing the binding sites and the effect of berbamine on the structure of bovine serum albumin. AB - Berbamine, a naturally occurring isoquinoline alkaloid extracted from Berberis sp., is the active constituent of some Chinese herbal medicines and exhibits a variety of pharmacological activities. The effects of berbamine on the structure of bovine serum albumin (BSA) were investigated by circular dichroism, fluorescence and absorption spectroscopy under physiological conditions. Berbamine caused a static quenching of the intrinsic fluorescence of BSA, and the quenching data were analyzed by application of the Stern-Volmer equation. There was a single primary berbamine-binding site on BSA with a binding constant of 2.577x10(4)Lmol(-1) at 298K. The thermodynamic parameters, enthalpy change (DeltaH(0)) and entropy change (DeltaS(0)) for the reaction were -76.5kJmol(-1) and -173.4Jmol(-1)K(-1) according to the van't Hoff equation. The results showed that the hydrogen bond and van der Waals interaction were the predominant forces in the binding process. Competitive experiments revealed a displacement of warfarin by berbamine, indicating that the binding site was located at Drug sites I. The distance r between the donor (BSA) and the acceptor (berbamine) was obtained according to the Forster non-radiation energy transfer theory. The results of three-dimensional fluorescence spectra, UV-vis absorption difference spectra and circular dichroism of BSA in the presence of berbamine showed that the conformation of BSA was changed. The results provide a quantitative understanding of the effect of berbamine on the structure of bovine serum albumin, providing a useful guideline for further drug design. PMID- 19185536 TI - Hypersomnia, REM sleep fragmentation and loss of motivation in a patient with hypothalamic lesions. PMID- 19185537 TI - The four diagnostic criteria for Restless Legs Syndrome are unable to exclude confounding conditions ("mimics"). AB - BACKGROUND: Epidemiological survey studies have suggested that a large fraction of the adult population, from five to more than 10%, have symptoms of Restless Legs Syndrome (RLS). Recently, however, it has become clear that the positive predictive value of many questionnaire screens for RLS may be fairly low and that many individuals who are identified by these screens have other conditions that can "mimic" the features of RLS by satisfying the four diagnostic criteria. We noted the presence of such confounders in a case-control family study and sought to develop methods to differentiate them from true RLS. METHODS: Family members from the case-control study were interviewed blindly by an RLS expert using the validated Hopkins telephone diagnostic interview (HTDI). Besides questions on the four key diagnostic features of RLS, the HTDI contains open-ended questions on symptom quality and relief strategies and other questions to probe the character of provocative situations and modes of relief. Based on the entire HDTI, a diagnosis of definite, probable or possible RLS or Not-RLS was made. RESULTS: Out of 1255 family members contacted, we diagnosed 1232: 402 (32.0%) had definite or probable RLS, 42 (3.3%) possible RLS, and 788 (62.8%) Not-RLS. Of the 788 family members who were determined not to have RLS, 126 could satisfy all four diagnostic criteria (16%). This finding indicates that the specificity of the four criteria was only 84%. Those with mimic conditions were found to have atypical presentations whose features could be used to assist in final diagnosis. CONCLUSION: A variety of conditions, including cramps, positional discomfort, and local leg pathology can satisfy all four diagnostic criteria for RLS and thereby "mimic" RLS by satisfying the four diagnostic criteria. Definitive diagnosis of RLS, therefore, requires exclusion of these other conditions, which may be more common in the population than true RLS. Short of an extended clinical interview and workup, certain features of presentation help differentiate mimics from true RLS. PMID- 19185538 TI - The effect of gender on the prevalence of hypertension in obstructive sleep apnea. AB - BACKGROUND: Obstructive sleep apnea (OSA) causes systemic hypertension. However, there is conflicting data on the effect of gender on susceptibility to hypertension in OSA. Some show no gender differences in the prevalence of hypertension while others report either female or male propensity to have hypertension in the context of OSA. The inconsistencies in the results appear to be due to lack of full range of sleep apnea severity and insufficient number of women in the studies. We examined the effect of gender on prevalent hypertension in a cohort with a large representation of females with OSA. METHODS: A cross sectional study of a large cohort of subjects referred for sleep disorder evaluation. RESULTS: The cohort comprised of 736 with OSA and 315 without OSA. OSA was defined as apnea-hypopnea index (AHI) of 5/h. There were 529 men and 207 women in the OSA group with mean+/-SD age of 50+/-13 and 51+/-14 years, respectively. The control group consisted of 154 men and 161 women with mean+/-SD age of 44+/-15 and 43+/-14 years, respectively. The AHI in the OSA group varied from 5 to 197 with a median of 24 and a mean of 36 (interquartile range: 11-53). Multiple logistic regression analyses, modeling the association between AHI and hypertension and considering other covariates, showed that odds of hypertension increased with increasing age, BMI, and AHI. There was evidence that men were at higher risk for hypertension than women OR 1.82 (95%CI 1.01, 3.20) at the highest quartile of BMI. CONCLUSIONS: We have shown that the prevalence of hypertension increases with increasing age and severity of OSA, and markedly obese men may have a nearly 2-fold greater risk for hypertension than women in this clinic based population. PMID- 19185539 TI - Measurement properties of the Medical Outcomes Study Sleep Scale in patients with fibromyalgia. AB - OBJECTIVE: Sleep problems are a common symptom of fibromyalgia (FM). The objective of this study was to evaluate the Medical Outcomes Study (MOS) Sleep Scale as a measure of FM-related sleep problems. METHODS: Analyses were based on data from the 1056 and 1077 studies, two randomized, double-blind, placebo controlled trials of pregabalin for adults with FM. MOS Sleep Scale scores of study patients were compared with United States normative scores using a one sample Z test. Subscale structure of the MOS was evaluated by confirmatory factor analyses, internal consistency was evaluated using Cronbach's alpha reliability coefficients. Estimated clinically important differences (CID) in MOS Sleep Disturbance subscale scores were evaluated using mixed-effects models of change in subscale scores as a function of the Patient Global Impression of Change (PGIC). RESULTS: 1056 and 1077 included 748 and 745 patients, respectively. Most patients were female (1056: 94.4%, 1077: 94.5%) and white (1056: 90.2%, 1077: 91.0%). Mean ages were 48.8 years (1056) and 50.1 years (1077). Baseline MOS Sleep Scale scores were statistically (P<0.001) and substantially poorer than general population values. The MOS subscale structure was confirmed in both studies at each assessment except at baseline in the 1056 study. Cronbach's alpha coefficients were acceptable, at least 0.70, for all multi-item scales at baseline and end-of-study assessments in both studies, with the exception of the Sleep Adequacy subscale at baseline. The estimated CID for the MOS Sleep Disturbance subscale was 7.9. CONCLUSIONS: The MOS Sleep Scale is an appropriate measure of FM-related sleep problems. These analyses provide the foundation for further use and evaluation of the MOS Sleep Scale in FM patients. PMID- 19185540 TI - The brain hypocretins and their receptors: mediators of allostatic arousal. AB - The hypocretins (abbreviated 'Hcrts' - also called 'orexins') are two neuropeptides secreted exclusively by a small population of neurons in the lateral hypothalamus. These peptides bind to two receptors located throughout the brain in nuclei associated with diverse cognitive and physiological functions. Initially, the brain Hcrt system was found to have a major role in the regulation of sleep/wake transitions. More recent studies indicate Hcrts may play a role in other physiological functions, including food intake, addiction, and stress. Taken together, these studies suggest a general role for Hcrts in mediating arousal, especially when an organism must respond to unexpected stressors and challenges in the environment. PMID- 19185541 TI - How can your parasites become your allies? AB - Although parasitic infection is usually detrimental, it can be beneficial to the host in some situations. Parasites could help their host by providing a new function or modifying one of the host's life-history traits. We argue that the evolution towards a lasting mutualistic relationship would be more likely when parasites endow hosts with new abilities rather than alter a trait because hosts are less likely to evolve a new capability on their own than adjust their life history by microevolutionary steps. Furthermore, we underline how evolved dependence--the host's loss of ability to live alone owing to a long history of evolution in the presence of its parasites--has shaped contemporary mutualistic relationships. PMID- 19185542 TI - Epidemiology of type 1 diabetes and what animal models teach us about the role of viruses in disease mechanisms. AB - There is a consensus among epidemiologists that the worldwide incidence rate of type 1 diabetes has been rising in recent decades. The cause of this rise is unknown, but epidemiological studies suggest the involvement of environmental factors, and viral infections in particular. Data demonstrating a cause-and effect relationship between microbial infections and type 1 diabetes and how viruses may cause disease in humans are currently lacking. However, new evidence from animal models supports the hypothesis that viruses induce disease via mechanisms linked with innate immune upregulation. In the BioBreeding Diabetes Resistant rat, infection with a parvovirus induces islet destruction via upregulation of the toll-like receptor 9 (TLR9) signaling pathway. Data from mouse models of diabetes implicate TLR2, TLR3, and TLR7 in the disease process. Understanding the link between environmental agents and innate immune pathways involved in early stages of diabetes may advance the design of immune interventions to prevent disease in genetically susceptible individuals. PMID- 19185543 TI - Mannose binding lectin gene polymorphisms and associated auto-immune diseases in type 1 diabetes Brazilian patients. AB - In our study we investigated the possible role of MBL2 functional single nucleotide polymorphisms (SNPs) in the augmented susceptibility to develop other autoimmune diseases in presence of type 1 diabetes (T1D) in a group of Brazilian patients. Patients were stratified for the presence of autoimmune diseases known to be associated with T1D, such as autoimmune thyroid disease (AITD) and celiac disease (CD), and compared with healthy controls (HC). Our findings suggest that MBL2 functional SNPs are more closely related to AITD than to T1D, being MBL2 SNPs frequencies in T1D patients not affected by AITD comparable to the HC ones, while significantly different between AITD patients and patients not affected by the disease. Thus, the association between MBL2 polymorphisms and T1D that we previously reported, seems to result from the stronger association of MBL2 SNPs with another autoimmune disease, the AITD, frequently associated with T1D. PMID- 19185544 TI - siRNA-mediated knockdown of the NR1 subunit gene of the NMDA receptor attenuates formalin-induced pain behaviors in adult rats. AB - NMDA receptors in the spinal cord dorsal horn (SCDH) mediate some inflammatory pain behaviors. Here, we used rAAV vectors expressing an active small interfering RNA (siRNA) (vector 6) targeting the essential NR1 subunit of the NMDA receptor or a mismatch siRNA (vector MM-6) sequence to determine the consequences of RNAi mediated knockdown of NR1 expression on NMDA receptor levels and formalin-induced pain behaviors in adult rats. Three weeks after intraparenchymal administration of the vector 6 into the right lumbar SCDH, NR1 mRNA and protein levels were significantly reduced (P < .01) in the ipsilateral SCDH compared with the contralateral SCDH but not in vector MM-6 or non-vector control animals. Formalin induced phase 2 nociceptive response was significantly reduced (P < .05) in vector 6 animals compared with controls. Although neither vector affected normal mechanical threshold, vector 6 provided protection from the mechanical allodynia seen in controls at 24 hours after intraplantar formalin. Vector 6 also prevented the increase in phosphorylated NR1 levels seen in the ipsilateral SCDH of control rats 45 minutes after formalin. These results indicate that vector-derived siRNAs can effectively produce spatial knockdown of NR1 gene expression, and this knockdown selectively attenuates in vivo NMDA receptor-mediated formalin behaviors and NR1 phosphorylation in the rat. PERSPECTIVE: This study reveals that a single administration of an siRNA-expressing viral vector produces significant knockdown of the NR1 gene in the SCDH of adult rats. This preclinical study demonstrates the use of RNAi to target the expression of genes mediating pain and the therapeutic potential of this approach. PMID- 19185545 TI - Individual differences in pain sensitivity: measurement, causation, and consequences. AB - Not only are some clinical conditions experienced as more painful than others, but the variability in pain ratings of patients with the same disease or trauma is enormous. Available evidence indicates that to a large extent these differences reflect individual differences in pain sensitivity. Pain sensitivity can be estimated only through the use of well-controlled experimental pain stimuli. Such estimates show substantial heritability but equally important environmental effects. The genetic and environmental factors that influence pain sensitivity differ across pain modalities. For example, genetic factors that influence cold pressor pain have little impact on phasic heat pain and visa versa. Individual differences in pain sensitivity can complicate diagnosis, among other reasons because low sensitivity to pain may delay self-referral. Inclusion of patients with reduced pain sensitivity can attenuate treatment effects in clinical trials, unless controlled for. Measures of pain sensitivity are predictive of acute postoperative pain, and there is preliminary evidence that heightened pain sensitivity increases risk for future chronic pain conditions. At this time, however, it is unclear which experimental pain modalities should be used as predictors for future pain conditions. Careful assessment of each individual's pain sensitivity may become invaluable for the prevention, evaluation, and treatment of pain. PERSPECTIVE: Large individual differences in pain sensitivity can complicate diagnosis and pain treatment and can confound clinical trials. Pain sensitivity may also be of great importance for the development of clinical pain. Thus, assessment of pain sensitivity may be relevant for the prevention, evaluation, and treatment of acute and chronic pain. PMID- 19185547 TI - The medial forebrain bundle mediates cardiovascular responses to electrical stimulation of the medial prefrontal cortex. AB - The medial prefrontal cortex (MPFC) is involved in cardiovascular control. MPFC electrical stimulation has been reported to cause depressor and bradycardic responses in anesthetized rats. Although the pathway involved is yet unknown, there is evidence indicating the existence of a relay in the lateral hypothalamus (LH). The medial forebrain bundle (MFB) that courses in the lateral portion of the LH carries the vast majority of telencephalic afferent as well efferent projections, including those from the MPFC. To evaluate if the hypotensive pathway originating in the MPFC courses the MFB, we studied the effect of coronal or sagittal knife cuts through the LH and other brain areas on the cardiovascular responses to MPFC electrical stimulation. Knife cuts were performed using blades 1 to 6 mm wide. Results indicate that the neural pathway descending from the MFB decussates early in the vicinity of MPFC, crossing the midline within the corpus callosum and yielding two descending pathways that travel rostro-caudally in the lateral portion of the LH, within the MFB. The decussation was confirmed by histological analysis of brain sections processed after the injection of biotinilated dextran amine in the site of the stimulation in the MPFC. Because knife cuts through the LH ipsilateral had minimal effects on the cardiovascular responses and knife cuts performed contralateral to the stimulated MPFC had no effect on the response to MPFC stimulation, data indicate that the contralateral limb of the pathway may be only activated as an alternative pathway when the ipsilateral pathway is blocked. PMID- 19185546 TI - Acid-sensing ion channel 3 expression in mouse knee joint afferents and effects of carrageenan-induced arthritis. AB - Arthritis is associated with decreases in local pH. Of the acid-sensing ion channels (ASIC), ASIC3 is most sensitive to such a pH change, abundantly expressed in dorsal root ganglion (DRG), and critical for the development of secondary hyperalgesia. The purpose of this study was to investigate the upregulation of ASIC3, using an acute arthritic pain model in mice. We examined ASIC3 expression in DRG neurons innervating the knee joint with and without carrageenan-induced arthritis by means of retrograde labeling and immunohistochemistry. We also examined the difference of DRG phenotype between ASIC3+/+ and ASIC3-/- mice. ASIC3 immunoreactivity was present in 31% of knee joint afferents and dominantly in small cells. After joint inflammation, ASIC3 immunoreactive neurons significantly increased in number by 50%. Calcitonin gene related peptide (CGRP) increased similarly in both ASIC3+/+ and ASIC3-/- mice. Soma size distribution of ASIC3-immunoreactive neurons without CGRP expression was shifted to smaller-diameter neurons. Our results suggest that ASIC3 plays an important role in acute arthritic pain. Specifically, we propose that ASIC3 upregulation along with CGRP and phenotypic change in ASIC3-immunoreactive neurons without CGRP are responsible for the development of secondary hyperalgesia after carrageenan-induced arthritis. PERSPECTIVE: This article shows that ASIC3 is upregulated along with CGRP in knee joint afferents and that there is a phenotypic change in ASIC3-immunoreactive nonpeptidergic neurons in an animal model of acute arthritis. Understanding the basic neurobiology after acute arthritis could lead to future new pharmacological management of arthritis. PMID- 19185548 TI - The role of novel genes rrp1(+) and rrp2(+) in the repair of DNA damage in Schizosaccharomyces pombe. AB - We identified two predicted proteins in Schizosaccharomyces pombe, Rrp1 (SPAC17A2.12) and Rrp2 (SPBC23E6.02) that share 34% and 36% similarity to Saccharomyces cerevisiae Ris1p, respectively. Ris1p is a DNA-dependent ATP-ase involved in gene silencing and DNA repair. Rrp1 and Rrp2 also share similarity with S. cerevisiae Rad5 and S. pombe Rad8, containing SNF2-N, RING finger and Helicase-C domains. To investigate the function of the Rrp proteins, we studied the DNA damage sensitivities and genetic interactions of null mutants with known DNA repair mutants. Single Deltarrp1 and Deltarrp2 mutants were not sensitive to CPT, 4NQO, CDPP, MMS, HU, UV or IR. The double mutants Deltarrp1 Deltarhp51 and Deltarrp2 Deltarhp51 plus the triple Deltarrp1 Deltarrp2 Deltarhp51 mutant did not display significant additional sensitivity. However, the double mutants Deltarrp1 Deltarhp57 and Deltarrp2 Deltarhp57 were significantly more sensitive to MMS, CPT, HU and IR than the Deltarhp57 single mutant. The checkpoint response in these strains was functional. In S. pombe, Rhp55/57 acts in parallel with a second mediator complex, Swi5/Sfr1, to facilitate Rhp51-dependent DNA repair. Deltarrp1 Deltasfr1 and Deltarrp2 Deltasfr1 double mutants did not show significant additional sensitivity, suggesting a function for Rrp proteins in the Swi5/Sfr1 pathway of DSB repair. Consistent with this, Deltarrp1 Deltarhp57 and Deltarrp2 Deltarhp57 mutants, but not Deltarrp1 Deltasfr1 or Deltarrp2 Deltasfr1 double mutants, exhibited slow growth and aberrations in cell and nuclear morphology that are typical of Deltarhp51. PMID- 19185549 TI - Normalization strategies for metabonomic analysis of urine samples. AB - Unlike plasma and most biological fluids which have solute concentrations that are tightly controlled, urine volume can vary widely based upon water consumption and other physiological factors. As a result, the concentrations of endogenous metabolites in urine vary widely and normalizing for these effects is necessary. Normalization approaches that utilized urine volume, osmolality, creatinine concentration, and components that are common to all samples ("total useful MS signal") were compared in order to determine which strategies could be successfully used to differentiate between dose groups based upon the complete endogenous metabolite profile. Variability observed in LC/MS results obtained from targeted and non-targeted metabonomic analyses was highly dependent on the strategy used for normalization. We therefore recommend the use of two different normalization techniques in order to facilitate detection of statistically significant changes in the endogenous metabolite profile when working with urine samples. PMID- 19185550 TI - Solid-phase microextraction using poly(pyrrole) film and liquid chromatography with UV detection for analysis of antidepressants in plasma samples. AB - Poly(pyrrole) (PPY) coating was prepared on a stainless-steel (SS) wire for solid phase microextraction (SPME) by electrochemical deposition (cyclic voltammetric). The PPY was evaluated by analyzing new-generation antidepressants (mirtazapine, citalopram, paroxetine, duloxetine, fluoxetine, and sertraline) in plasma sample by SPME and liquid chromatography with UV detection (LC-UV). The effect of electrolyte solution (lithium perchlorate or tetrabutylammonium perchlorate) and the number of cycles (50, 100 or 200) applied during the polymerization process on the SPME performance was evaluated. Important factors in the optimization of SPME efficiency such as extraction time, temperature, pH, influence of plasma proteins on sorption mechanisms, and desorption conditions are discussed. The SPME-PPY/LC method showed to be linear in concentrations ranging from the limit of quantification (LOQ) to 1200 ng mL(-1). The LOQ values range from 16 to 25 ng mL(-1). The inter-day precision of the SPME-PPY/LC method presented coefficient of variation (CV) lower than 15%. Based on analytical validation results, the SPME-PPY/LC methodology showed to be adequate for antidepressant analysis, from therapeutic to toxic levels. In order to evaluate the proposed method for clinical use, the SPME-PPY/LC method was applied to the analysis of plasma samples from elderly depressed patients. PMID- 19185551 TI - Neonatal intrahepatic cholestasis caused by citrin deficiency: clinical and laboratory investigation of 13 subjects in mainland of China. AB - BACKGROUND: Neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD) is a novel inborn error of metabolism due to dysfunction of citrin protein, and much more information about this new disease is still needed for its clinical management. AIMS: To investigate in detail the clinical and laboratory features of NICCD. PATIENTS: 13 NICCD subjects in mainland of China diagnosed in our department since 2006. METHODS: The anthropometric parameters of the patients at birth were compared with controls, representative biochemical changes and metabolome findings were investigated cross-sectionally, and mutations in the causative gene SLC25A13 were analyzed by protocols established previously. RESULTS: The patients showed reduced birth weight, length and ponderal index. Main clinical manifestations consisted of jaundice, hepato/hepatosplenomegaly and steatohepatosis on ultrasonography. Biochemical analysis revealed intrahepatic cholestasis, delayed switch of AFP to albumin, and elevated triglyceride, total cholesterol and LDL-cholesterol together with reduced HDL-cholesterol. Metabolome findings included co-existence of markers for galactosemia and tyrosinemia in urine, and elevated Cit, Met, Thr, Tyr, Lys, Arg and Orn in blood. Mutations of 851-854del, IVS6+5G>A, 1638-1660dup, A541D, IVS16ins3kb, R319X and G333D were detected in the gene SLC25A13. CONCLUSIONS: The diagnosis of NICCD cannot be established based just on the numerous but non-specific clinical manifestations and biochemical changes. The relatively specific metabolome features provide valuable tools for its screening and diagnosis, while SLC25A13 mutation analysis should be taken as one of the reliable tools for the definitive diagnosis. The body proportionality at birth, steatohepatosis on ultrasonography, delayed switch of AFP to albumin, dyslipidemia pattern, urinary metabolome features and the novel mutation G333D expanded the clinical spectrum of NICCD. PMID- 19185552 TI - Eosinophilic colitis. PMID- 19185553 TI - Liver HCV-antigens and steatosis in chronic hepatitis C: role of different genotypes. AB - BACKGROUND: HCV infection is frequently associated with liver steatosis. AIMS AND METHODS: We studied 126 frozen liver HCV positive specimens (genotype-3=27) without any features of metabolic syndrome, searching for a correlation between the number of HCV infected hepatocytes and the presence, amount and distribution of steatosis in relation to different genotypes. RESULTS: Mean steatosis score was higher in genotype-3 with respect to non-3 (1.11 vs 0.66, p=0.022). HCV antigens were detected by an immunoperoxidase technique in 91/126 (72.2%) cases. A significant correlation between the number of HCV-antigen positive cells and the degree of liver steatosis was observed in genotype-3 (p=0.01) but not in non 3 patients, matched for sex and age. Steatotic cells usually outnumbered HCV infected cells. Steatosis was observed both in HCV-antigen positive and negative hepatocytes, and HCV-antigens were detected in both hepatocytes with and without steatosis: while no lobular codistribution was found in genotype non-3, in genotype-3 steatosis and HCV-antigens were usually found in the same areas. CONCLUSION: Our data support the role of HCV-antigen liver expression in the pathogenesis of steatosis in genotype-3, however, since the presence of HCV antigens is not directly related to steatosis within single hepatocytes, an indirect mechanism might be operative too. PMID- 19185554 TI - Alcohol and coffee drinking and smoking habit among subjects with HCV infection. AB - BACKGROUND/AIMS: The aims were to estimate among patients with hepatitis C virus (HCV) infection the prevalence of alcohol and coffee intake and smoking habit, the reliability of these self-reported data and the possible change of patients' habit after their first contact with a Viral Hepatitis Service. METHODS: 229 patients were initially interviewed personally at the Viral Hepatitis Service and after 6 months they were re-interviewed by phone in regard to their alcohol, coffee drinking and smoking habits. RESULTS: Alcohol drinkers were 55.5% of males and 35.3% of females. Most subjects drank coffee daily, both men (90.0%) and women (84.9%). The proportion of current smokers was higher in males (43.6%) than females (26.9%). We found a fair to good reliability of self-reported data regarding patients' habits, alcohol and coffee intake, and number of cigarettes smoked daily. We observed a statistically significant decrease in alcohol and coffee intake and cigarettes smoked between baseline and follow-up interviews. CONCLUSION: We found a fairly high proportion of HCV-infected patients who regularly drink alcohol and coffee beverages and smoke cigarettes, especially among males. The reliability of self-reported data on these habits seems satisfactory. More decisive action to modify these habits, especially alcohol intake, is required in these patients. PMID- 19185555 TI - A modified TNM-based Japan Integrated Score combined with AFP level may serve as a better staging system for early-stage predominant hepatocellular carcinoma patients. AB - BACKGROUND: Combinations of Child-Pugh classification and Liver Cancer Study Group of Japan/Tumor-Node-Metastasis (LCSGJ/TNM) have been reported as Japan Integrated Staging (JIS). We previously modified the 6th AJCC/TNM to serve as a better staging system than the 5th and 6th AJCC/TNM. AIMS: To develop a modified TNM-based JIS to predict the survival of hepatocellular carcinoma (HCC) patients more accurately. METHODS: 3764 HCC patients were enrolled from 1986 to 2002 (2882 patients from 1986 to 2000 and 882 patients from 2001 to 2002). We compared the performance of original JIS, modified TNM-based JIS, modified TNM-based JIS combined alpha-fetoprotein (AFP), BCLC, and CLIP. Lower Akaike information criteria (AIC) values indicated better discriminatory abilities. RESULTS: AIC value was lowest in CLIP during all periods. However, during 2001-2002, when early-stage HCC patients were predominant, AIC value was lowest when modified TNM based JIS combined AFP was used. CONCLUSION: The CLIP system provided the best prognostic stratification in the present cohort of HCC patients who were mainly at late stages. However, early detection of HCCs has become more common in Taiwan in recent years, which has led to the predominance of early-stage HCC patients. Therefore, modified TNM-based JIS combined AFP may now be the most applicable system in recent years. PMID- 19185556 TI - The role of oesophageal diversion and exclusion in the management of oesophageal perforations. AB - AIM: Persistent sepsis from an oesophageal perforation has a near 100% mortality. We describe our experience with early oesophageal diversion and exclusion for patients in-extremis. METHODS: A retrospective review of oesophageal perforations was performed between 2000 and 2007. There were five cases Boerhaaves and one case of iatrogenic perforation that required oesophageal diversion and exclusion. 4 males, 2 females with a mean age of 67.6 (58-72) years. RESULTS: The primary procedure was performed within 24h in four patients; the other two were after 3 and 10 days. The intensive care unit (ITU) stay was a median of 25 days. Mortality rate was 50%. Median length of stay for the survivors was 60 days. Three patients underwent a successful colonic interposition in our unit after 6 months. CONCLUSION: Exclusion and diversion procedures are required in very rare circumstances. In conditions of persistent leak and continuing sepsis or those patients not fit to undergo a major procedure they could be lifesaving if performed early. As it is a relatively easy and quick procedure it should be considered early as a 2nd line management option. PMID- 19185557 TI - Periosteal chondroma of the clavicle: case report and review of the literature. AB - Periosteal chondromas (juxtacortical chondromas), are slow growing, rare cartilaginous lesion that arises adjacent to the cortex beneath the periosteum. They occur more in males in their 20s. We report a rare case of periosteal chondroma arising from the left clavicle of a 56 year old male. We could only find one reported case in the English literature of periosteal chondroma arising from the research and to the best of our knowledge, this is the second reported case. PMID- 19185558 TI - Monitoring of microcirculation in free transferred musculocutaneous lattisimus dorsi flaps by confocal-laser-scanning microscopy--a promising noninvasive methodical approach. PMID- 19185559 TI - The unilocular fat-cell graft. AB - Autologous fat grafting has become a common procedure for augmenting soft tissue. However, there are still some problems with resorption and cyst formation after injection, which mainly arise from insufficient nourishment of the transplanted fat tissues. In this study, using a mouse model, we enzymatically digested fat tissues into unilocular fat cells, and then transplanted the unilocular fat cells by sub-dermal injection to allow the transplanted cells to easily spread within the injected area. Fat tissue was harvested from a green fluorescent protein transgenic mouse (C57BL/6), and the optimal digestion time was determined to be 30 min. The fat cells were then injected into the sub-dermal layer of the head skin of a C57BL/6 mouse. As a control, minced fat without digestion was also injected. The animals were sacrificed immediately after injection and on days 1, 2 and 3, as well as at weeks 1, 2 and 4 after injection, and the recipient skins were collected for microscopic observation. The unilocular fat cells were observed to spread in a solitary manner among the recipient tissues, and no necrotic areas or cysts were observed. The minced-fat-graft control showed central necrosis in the transplanted region. In addition, the minced fat tissue needed to be injected with an 18-gauge syringe, but the unilocular fat cells could be injected with a 26-gauge syringe. Thus, the unilocular fat-cell graft was determined to be a superior alternative to conventional fat grafts. PMID- 19185560 TI - Alendronate/vitamin D3 70 mg/2800 IU with and without additional 2800 IU vitamin D3 for osteoporosis: results from the 24-week extension of a 15-week randomized, controlled trial. AB - Although vitamin D supplementation is a fundamental part of osteoporosis treatment, many patients do not regularly take adequate amounts. A once-weekly (OW) alendronate (ALN) preparation that includes 2800 IU of vitamin D3 in a single combination tablet (ALN+D2800) is available for treating patients and ensuring intake of vitamin D that is consistent with existing guidelines. This randomized, double-blind study extension was conducted to evaluate the safety and tolerability of ALN+D2800 and ALN+D2800 plus an additional 2800 IU vitamin D3 single tablet supplement (ALN+D5600) administered for 24 weeks in men and postmenopausal women with osteoporosis previously treated OW for 15 weeks with either ALN or ALN+D2800. The primary endpoint was the proportion of participants who developed hypercalciuria (defined as a 24-hour urine calcium >300 mg in women or >350 mg in men and an increase of >25% versus randomization baseline) at week 39. The key secondary endpoint was the proportion of participants with vitamin D insufficiency (serum 25(OH)D <15 ng/mL [37.4 nmol/L]) at the end of the study. Hypercalciuria incidence (4.2% [ALN+D5600] vs. 2.8% [ALN+D2800]), did not differ between groups (p = 0.354). No participants developed hypercalcemia. Among the participants with vitamin D insufficiency at the week 0 baseline, the prevalence of insufficiency at the end of the study was reduced by 92% in the ALN+D5600 group and by 86% in the ALN+D2800 group. The incidences of clinical adverse experiences, including drug-related adverse experiences, were similar in both groups. In subjects previously treated with ALN+D2800 for 15 weeks, the addition of 2800 IU D3 for 24 weeks did not produce hypercalcemia nor increase the risk of hypercalciuria. PMID- 19185562 TI - An alternative assay to discover potential calmodulin inhibitors using a human fluorophore-labeled CaM protein. AB - This article describes the development of a new fluorescent-engineered human calmodulin, hCaM M124C-mBBr, useful in the identification of potential calmodulin (CaM) inhibitors. An hCaM mutant containing a unique cysteine residue at position 124 on the protein was expressed, purified, and chemically modified with the fluorophore monobromobimane (mBBr). The fluorophore-labeled protein exhibited stability and functionality to the activation of calmodulin-sensitive cAMP phosphodiesterase (PDE1) similar to wild-type hCaM. The hCaM M124C-mBBr is highly sensitive to detecting inhibitor interaction given that it showed a quantum efficiency of 0.494, approximately 20 times more than the value for wild-type hCaM, and a large spectral change ( approximately 80% quenching) when the protein is in the presence of saturating inhibitor concentrations. Two natural products previously shown to act as CaM inhibitors, malbrancheamide (1) and tajixanthone hydrate (2), and the well-known CaM inhibitor chlorpromazine (CPZ) were found to quench the hCaM M124C-mBBr fluorescence, and the IC(50) values were comparable to those obtained for the wild-type protein. These results support the use of hCaM M124C-mBBr as a fluorescence biosensor and a powerful analytical tool in the high throughput screening demanded by the pharmaceutical and biotechnology industries. PMID- 19185561 TI - Molecular cloning and characterization of heat shock protein 70 from Trichinella spiralis. AB - A cDNA encoding heat shock protein 70 of Trichinella spiralis (Ts-Hsp70) was identified by immunoscreening the adult T. spiralis cDNA library with rabbit antisera against T. spiralis adult extracts. The open reading frame of Ts-Hsp70 cDNA encoded a 623-amino acid peptide with a predicted molecular weight of 68.7kDa, which shares a high degree of sequence conservation with HSP70s from other parasites. Recombinant Ts-Hsp70 was expressed in Escherichia coli and purified with nickel column chromatography. Western blot analysis showed that recombinant Ts-Hsp70 could be recognized not only by trichinellosis patient sera, but also by T. spiralis-infected sera from rabbits, swine, and mice. Mice vaccinated with recombinant Ts-Hsp70 formulated with Freund's adjuvant exhibited strong humoral immune responses indicated by high titer of IgG antibody and significant muscle larval reduction (37%) after being challenged with T. spiralis larvae. The present results indicate that Ts-Hsp70 is a possible candidate vaccine against T. spiralis infection. PMID- 19185563 TI - Single-step synthesis of 4-nitrophenyl ferulate for spectrophotometric assay of feruloyl esterases. AB - We report here, for the first time, a single-step method for the synthesis of 4 nitrophenyl ferulate (4NPF), a spectrophotometric substrate for the assay of feruloyl esterases by dehydrative coupling of ferulic acid and 4-nitrophenol. PMID- 19185564 TI - Apoptosis, autophagy, accelerated senescence and reactive oxygen in the response of human breast tumor cells to adriamycin. AB - Although the primary response to Adriamycin (doxorubicin) in p53 mutant MDA-MB231 and p53 null MCF-7/E6 breast tumor cells is apoptotic cell death, the residual surviving population appears to be in a state of senescence, based on cell morphology, beta galactosidase staining, induction of p21(waf1/cip1) and down regulation of cdc2/cdk1. Suppression of apoptosis in MDA-MB231 and MCF-7/E6 cells treated with Adriamycin using the broad spectrum caspase inhibitor, zvad-Fmk, results in substantial induction of autophagy. Overall sensitivity to Adriamycin, measured by clonogenic survival, is not altered in the cells undergoing autophagy, consistent with autophagy contributing to cell death in response to Adriamycin. The free radical scavengers, glutathione and N-acetyl cysteine attenuate the accelerated senescence response to Adriamycin in MCF-7 cells as well as in MDA-MB231 and MCF-7/E6 cells, but protect primarily the MCF-7 cells, indicating that reactive oxygen is unlikely to be directly responsible for Adriamycin toxicity in breast tumor cells. Expression of caspase 3 or induced expression of c-myc in MCF-7 cells fails to abrogate accelerated senescence induced by Adriamycin. Taken together, these studies suggest that accelerated senescence induced by Adriamycin is similar in cells with wild type p53 and in cells lacking functional p53 with regard to the upregulation of p21(waf1/cip1), down regulation of cdc2 and the involvement of reactive oxygen species. Furthermore, accelerated senescence, autophagy and apoptosis all appear to be effective in suppressing self-renewal capacity in breast tumor cells exposed to Adriamycin. PMID- 19185565 TI - Carbon tetrachloride-induced liver damage in asialoglycoprotein receptor deficient mice. AB - The asialoglycoprotein (ASGP) receptor is an abundant hepatocyte-specific receptor involved in receptor-mediated endocytosis. This receptor's abundance and function is decreased by chronic ethanol administration prior to the appearance of pathology such as necrosis or inflammation. Hence, this study aimed to determine if ASGP receptor function is required to protect against liver injury by utilizing a knockout mouse model lacking functional ASGP receptor in the setting of carbon tetrachloride (CCl(4)) hepatotoxicity. Briefly, ASGP receptor deficient (RD) mice and wild-type (WT) mice were injected with 1ml/kg body weight of CCl(4). In the subsequent week, mice were monitored for liver damage and pathology (aspartate transaminase (AST), alanine transaminase (ALT) and light microscopy). The consequences of CCl(4) injection were examined by measuring alpha-smooth muscle actin (alpha-SMA) deposition, contents of malondialdehyde and the percentage of apoptotic hepatocytes. After CCl(4) injection, RD mice showed increased liver pathology together with significantly increased activities of AST and ALT compared to that in WT mice. There were also significantly more apoptotic bodies, lipid peroxidation and deposition of alpha-SMA in RD mice versus WT mice following CCl(4) injection. Since these two mouse strains only differ in whether or not they have the ASGP receptor, it can be concluded that proper ASGP receptor function exerted a protective effect against CCl(4) toxicity. Thus, receptor mediated endocytosis by the ASGP receptor could represent a novel molecular mechanism that is responsible for subsequent liver health or injury. PMID- 19185566 TI - Role of carboxylesterase 1 and impact of natural genetic variants on the hydrolysis of trandolapril. AB - Carboxylesterase 1 (CES1) and carboxylesterase 2 (CES2) are the major hydrolytic enzymes responsible for the metabolism of numerous therapeutic agents as well as endogenous substrates. CES1 and CES2 differ distinctly in their substrate specificity and tissue distribution. In this study, we investigated the role of CES1 and CES2 in converting the antihypertensive prodrug trandolapril to its more active form trandolaprilat, and determined the influence of two newly identified CES1 mutations p.Gly143Glu and p.Asp260fs on trandolapril metabolism. Western blot analysis demonstrated that CES1 is expressed in human liver microsomes (HLM) but not in human intestinal microsomes (HIM). In vitro incubation studies were conducted to contrast the enzymatic activity of HLM as well as HIM upon trandolapril hydrolysis. Trandolapril was rapidly hydrolyzed to its principal active metabolite trandolaprilat after incubation with HLM. In contrast, in HIM, where CES2 is predominantly expressed, incubations did not produce any detectable trandolapril hydrolysis. Furthermore, hydrolysis of trandolapril catalyzed by wild type (WT) and mutant CES1 were assessed utilizing transfected Flp-In-293 cells stably expressing WT CES1 and two variants. WT CES1 efficiently hydrolyzed trandolapril to trandolaprilat with V(max) and K(m) values of 103.6+/-2.2 nmole/min/mg protein and 639.9+/-32.9muM, respectively. However, no appreciable trandolapril hydrolysis could be found after incubation with both p.Gly143Glu and p.Asp260fs variants. Thus, trandolapril appears to be a CES1 selective substrate while CES2 exerts little to no catalytic activity towards this compound. CES1 mutations p.Gly143Glu and p.Asp260fs are essentially dysfunctional enzymes with regard to the conversion of trandolapril to its more active metabolite trandolaprilat. PMID- 19185567 TI - Neural mechanism of intertemporal choice: from discounting future gains to future losses. AB - Intertemporal choice, the tradeoff among outcomes occurring at different points in time, involves not only benefit options but also those associated with cost. Previous neuroimaging studies have primarily focused on discounting future gains; thus the neural mechanism underlying discounting future losses remains unidentified. Using event-related functional magnetic resonance imaging, we comprehensively investigated the neural mechanism of temporal discounting using two decision-making tasks with a symmetric pattern of gains and losses. Our results revealed that the lateral prefrontal and posterior parietal areas were activated in discounting both future gains and future losses, but their activations were stronger when discounting losses. Moreover, we found that the insula, thalamus and dorsal striatum were more activated during intertemporal choices involving losses, suggesting that the enhanced sensitivity to losses may be driven by negative emotions. In addition, whereas the posterior cingulate cortex and medial prefrontal cortex were activated when the choices included immediate options, extra regions including the anterior cingulate cortex, insula and superior frontal gyrus were preferentially activated when the choices involved immediate losses. Taken together, our findings suggest that a fronto parietal network supports the common discounting process, and more importantly, discounting future losses and gains occurs asymmetrically in the brain. We speculate that this may provide a neural basis for the phenomenon that future losses are discounted less steeply than future gains. PMID- 19185568 TI - Changes in acetylcholinesterase (AchE) activity in lymphocytes and whole blood in acute lymphoblastic leukemia patients. AB - BACKGROUND: Acute lymphoblastic leukemia (ALL) is a type of cancer that affects lymphocytes and it is the most common form of cancer in children. Acetylcholinesterase (AChE) is well known as having non-cholinergic functions and has been detected in the blood and plasma of humans including in lymphocytes. Thus, we investigated whole blood and lymphocyte AChE activity in patients with ALL. METHODS: This study was performed on 72 children with ALL divided into 4 groups: newly diagnosed, remission induction, remission maintenance and out-of treatment and one control group of 50 healthy subjects. We determined AChE activity in whole blood and lymphocytes of these patients. RESULTS: Results demonstrated that whole blood AChE activity was enhanced in the newly diagnosed group and reduced in the remission induction and remission maintenance groups in relation to the control group. For lymphocyte AChE activity we found an increase in the newly diagnosed group and a decrease in the remission induction group in relation to the control. CONCLUSIONS: These results suggest that AChE activity was altered in ALL patients. This fact may be related with the essential role played by AChE in the development of hematological disease and its contribution to the regulation of immune function. PMID- 19185570 TI - Exogenous reelin prevents granule cell dispersion in experimental epilepsy. AB - Temporal lobe epilepsy (TLE) is often accompanied by granule cell dispersion (GCD), a migration defect of granule cells in the dentate gyrus. We have previously shown that a decrease in the expression of reelin, an extracellular matrix protein important for neuronal positioning, is associated with the development of GCD in TLE patients. Here, we used unilateral intrahippocampal injection of kainate (KA) in adult mice which is also associated with GCD formation and a decrease of reelin expression. In this mouse epilepsy model we aimed to prevent GCD development by the application of exogenous reelin. As a prerequisite we analyzed whether the reelin signaling transduction cascade was preserved in the KA-injected hippocampus. Using in situ hybridization and Western blot analysis we found that the expression of the reelin signaling components, apolipoprotein E receptor 2, the very-low-density lipoprotein receptor and the intracellular adaptor protein disabled 1, was maintained in dentate granule cells after KA injection. Next, recombinant reelin was infused into the KA-injected hippocampus by osmotic minipumps over a period of 2 weeks. Quantitative analysis of granule cell layer width revealed a significant reduction of GCD in reelin treated, but not in saline-infused animals when compared to KA injection alone. Our findings highlight the crucial role of reelin for the maintenance of granule cell lamination in the dentate gyrus of adult mice and show that a reelin deficiency is causally involved in GCD development. PMID- 19185569 TI - Elucidating timing and function of endothelin-A receptor signaling during craniofacial development using neural crest cell-specific gene deletion and receptor antagonism. AB - Cranial neural crest cells (NCCs) play an intimate role in craniofacial development. Multiple signaling cascades participate in patterning cranial NCCs, some of which are regulated by endothelin-A receptor (Ednra) signaling. Ednra(-/ ) embryos die at birth from severe craniofacial defects resulting from disruption of neural crest cell patterning and differentiation. These defects include homeotic transformation of lower jaw structures into upper jaw-like structures, suggesting that some cephalic NCCs alter their "identity" in the absence of Ednra signaling. To elucidate the temporal necessity for Ednra signaling in vivo, we undertook two strategies. We first used a conditional knockout strategy in which mice containing a conditionally targeted Ednra allele (Ednra(fl)) were bred with mice from the Hand2-Cre and Wnt1-Cre transgenic mouse strains, two strains in which Cre expression occurs at different time periods within cranial NCCs. In our second approach, we used an Ednra-specific antagonist to treat wild type pregnant mice between embryonic days E8.0 and E10.0, a time frame encompassing the early migration and proliferation of cranial NCCs. The combined results suggest that Ednra function is crucial for NCC development between E8.25 and E9.0, a time period encompassing the arrival of NCCs in the arches and/or early post-migratory patterning. After this time period, Ednra signaling is dispensable. Interestingly, middle ear structures are enlarged and malformed in a majority of Ednra(fl/fl);Wnt1-Cre embryos, instead resembling structures found in extinct predecessors of mammals. These observations suggest that the advent of Ednra signaling in cranial NCCs may have been a crucial event in the evolution of the mammalian middle ear ossicles. PMID- 19185571 TI - Over-expressed microRNA-27a and 27b influence fat accumulation and cell proliferation during rat hepatic stellate cell activation. AB - Hepatic stellate cells (HSCs) activation is an initial event in liver fibrosis. MicroRNAs (miRNAs) have been found to play essential roles in cell differentiation, proliferation, and fat metabolism. In this study, we showed that down-regulation of two over-expressed miRNAs, miR-27a and 27b allowed culture activated rat HSCs to switch to a more quiescent HSC phenotype, with restored cytoplasmic lipid droplets and decreased cell proliferation. Mechanistically, retinoid X receptor alpha was confirmed to be the target of miR-27a and 27b. These results indicated a new role and mechanism of miR-27a and 27b in regulating fat metabolism and cell proliferation during HSCs activation. PMID- 19185572 TI - Identification of the ice-binding face of a plant antifreeze protein. AB - The antifreeze protein of Lolium perenne, a perennial ryegrass, was previously modeled as a beta-roll with two extensive flat beta-sheets on opposite sides of the molecule. Here we have validated the model with a series of nine site directed steric mutations in which outward-pointing short side-chain residues were replaced by tyrosine. None of these disrupted the fold. Mutations on one of the beta-sheets and on the sides of the protein retained 70% or greater activity. Three mutations that clustered on the other flat surface lost up to 90% of their antifreeze activity and identify this beta-sheet as the ice-binding face. PMID- 19185573 TI - Intertwined dimeric structure for the SH3 domain of the c-Src tyrosine kinase induced by polyethylene glycol binding. AB - Here we report the first crystal structure of the SH3 domain of the cellular Src tyrosine kinase (c-Src-SH3) domain on its own. In the crystal two molecules of c Src-SH3 exchange their -RT loops generating an intertwined dimer, in which the two SH3 units, preserving the binding site configuration, are oriented to allow simultaneous binding of two ligand molecules. The dimerization of c-Src-SH3 is induced, both in the crystal and in solution, by the binding of a PEG molecule at the dimer interface, indicating that this type of conformations are energetically close to the native structure. These results have important implications respect to in vivo oligomerization and amyloid aggregation. PMID- 19185574 TI - The immunostimulatory effect of IL-1beta in vivo is blocked by antisense peptides complementary to the loop sequence 163-171. AB - Antisense (AS) peptides complementary to the beta-bulge surface loop VQGEESNDK (Boraschi loop) of the cytokine interleukin-1beta (IL-1beta) have been shown to bind IL-1beta at the Boraschi loop position, and to inhibit some of the IL-1beta mediated biological effects in vitro. Here we show that primary AS peptide FVITFFSLY inhibits IL-1beta-mediated immunostimulation in vivo in a dose dependent fashion, while inactive on IL-1beta-induced inflammation, an effect that takes place independently of the Boraschi loop. To the best of our knowledge, this is the first time that an AS peptide has been used successfully in vivo. PMID- 19185575 TI - Antisense oligonucleotides effectively inhibit the co-transcriptional splicing of a Candida group I intron in vitro and in vivo: Implications for antifungal therapeutics. AB - Self-splicing of group I intron from the 26S rRNA of Candida albicans is essential for maturation of the host RNA. Here, we demonstrated that the co transcriptional splicing of the intron in vitro was blocked by antisense oligonucleotides (AONs) targeting the P3-P7 core of the intron. The core-targeted AON effectively and specifically inhibited the intron splicing from its host RNA in living C. albicans. Furthermore, flow cytometry experiments showed that the growth inhibition was caused by a fungicidal effect. For the first time, we showed that an AON targeting the ribozyme core folding specifically inhibits the endogenous ribozyme splicing in living cells and specifically kills the intron containing fungal strains, which sheds light on the development of antifungal drugs in the future. PMID- 19185576 TI - Probing the orientation of yeast VDAC1 in vivo. AB - Voltage dependent anion channel (VDAC) is a vital ion channel in mitochondrial outer membranes and its structure was recently shown to be a 19 stranded beta barrel. However the orientation of VDAC in the membrane remains unclear. We probe here the topology and membrane orientation of yeast Saccharomyces cerevisiae in vivo. Five FLAG-epitopes were independently inserted into scVDAC1 and their surface exposure in intact and disrupted mitochondria detected by immunoprecipitation. Functionality was confirmed by measurements of respiration. Two epitopes suggest that VDAC (scVDAC) has its C-terminus exposed to the cytoplasm whilst two others are more equivocal and, when combined with published data, suggest a dynamic behavior. PMID- 19185577 TI - Sall4 regulates cell fate decision in fetal hepatic stem/progenitor cells. AB - BACKGROUND & AIMS: Fetal hepatic stem/progenitor cells, called hepatoblasts, differentiate into both hepatocytes and cholangiocytes. The molecular mechanisms regulating this lineage segmentation process remain unknown. Sall4 has been shown to be among the regulators of organogenesis, embryogenesis, maintenance of pluripotency, and early embryonic cell fate decisions in embryonic stem cells. The expression and functional roles of Sall4 during liver development have not been elucidated. We here provide their first description in hepatoblasts. METHODS: To investigate functions of Sall4 in fetal liver development, Dlk(+)CD45(-)Ter119(-) hepatoblasts derived from embryonic day 14 mouse livers were purified, and in vitro gain and loss of function analyses and in vivo transplantation analyses were performed using retrovirus- or lentivirus-mediated gene transfer. RESULTS: We demonstrated that Sall4 was expressed in fetal hepatoblasts but not adult hepatocytes. The expression level of Sall4 gradually fell during liver development. Overexpression of Sall4 in hepatoblasts significantly inhibited maturation induced by oncostatin M and extracellular matrix in vitro, as evidenced by morphologic changes and suppression of hepatic maturation marker gene expression. When bile duct-like structures were induced by collagen gel-embedded culture, overexpression of Sall4 markedly augmented size and number of cytokeratin19(+)-branching structures. Knockdown of Sall4 inhibited formation of these branching structures. With in vivo transplantation, Sall4 enhanced differentiation of cytokeratin19(+)-bile ducts derived from transplanted hepatoblasts. CONCLUSIONS: These results suggest that Sall4 plays a crucial role in controlling the lineage commitment of hepatoblasts not only inhibiting their differentiation into hepatocytes but also driving their differentiation toward cholangiocytes. PMID- 19185578 TI - Suppression of apoptosis, crypt hyperplasia, and altered differentiation in the colonic epithelia of bak-null mice. AB - BACKGROUND & AIMS: Members of the bcl-2 family of proteins are important determinants of cell fate. Bcl-2 and bcl-w have previously been identified as antiapoptotic members of this family that promote gastrointestinal epithelial cell survival. However, a proapoptotic family member that exerts important effects in the gastrointestinal tract has not yet been identified. We have therefore investigated intestinal epithelial apoptosis in bak-null mice. METHODS: Apoptosis, mitosis, differentiated cell composition, and cell number were assessed on a cell positional basis in the small intestinal and colonic epithelia of bak-null mice and their C57BL/6 wild-type counterparts. Apoptosis was induced by 1-Gy gamma-irradiation or 10mg/kg azoxymethane (AOM). Aberrant crypt foci were induced by 3 weekly injections of 10mg/kg AOM. RESULTS: The amount of spontaneous apoptosis in the colonic intercrypt table was reduced, and colonic crypt cell number and mitotic index were elevated in bak-null mice relative to C57BL/6 wild type mice. Bak-null colonic crypts contained more goblet cells and fewer endocrine cells than those from C57BL/6 mice. Fewer colonic epithelial apoptotic cells were observed after gamma-radiation and AOM in bak-null mice, and these mice also displayed greater numbers of colonic AOM-induced aberrant crypt foci. None of these parameters differed in the small intestinal epithelium of bak-null mice compared with C57BL/6. CONCLUSIONS: Bak prevents colonic crypt hyperplasia by regulating spontaneous apoptosis at the colonic intercrypt table region and also regulates damage-induced apoptosis in the colonic crypt. Deletion of bak in vivo results in altered colonic proliferation and differentiation, and causes increased susceptibility to colonic carcinogenesis. PMID- 19185579 TI - Analysis of CD8+ T-cell-mediated inhibition of hepatitis C virus replication using a novel immunological model. AB - BACKGROUND & AIMS: Virus-specific CD8+ T cells are required for the control of hepatitis C virus (HCV) infection. We investigated the extent to which different effector functions of CD8+ T cells contribute to the inhibition of viral replication. METHODS: We developed a novel immunologic model by stably transducing the HLA-A2 gene into the replicon system, matching the epitope sequence of the replicon to the sequence targeted by an HCV-specific CD8+ T-cell clone. Luciferase activity was then measured to quantitate HCV RNA replication. RESULTS: HCV-specific CD8+ T cells strongly inhibited viral replication, through cytolytic and noncytolytic mechanisms, in a dose-dependent manner. HCV replication was almost completely inhibited at an effector-to-target ratio of 1:1 with significant cytotoxicity; however, >95% viral inhibition occurred at ratios as low as 1:100. Importantly, no cytotoxicity was observed at low effector-to target ratios, indicating a dominant effect of noncytolytic effector functions that was confirmed by Transwell experiments. Neutralization experiments revealed that interferon gamma mediates the noncytolytic inhibition. CONCLUSIONS: Only a very few HCV-specific CD8+ T cells are required to inhibit HCV replication; inhibition occurs primarily by noncytolytic effector functions. PMID- 19185580 TI - The microRNA-30 family is required for vertebrate hepatobiliary development. AB - BACKGROUND & AIMS: The function of microRNA (miRNA) in liver development is unknown. To address this issue, we characterized miRNA expression in the embryonic mouse liver, performed functional miRNA analysis in zebrafish larvae, and identified novel hepatic miRNA targets. METHODS: Hepatic RNA isolated from mice at embryonic days 15.5, 18.5, and postnatal day 2 was hybridized to a mouse miRNA microarray. The microarray results were confirmed by Northern blot hybridization and quantitative reverse-transcription polymerase chain reaction. The spatial distribution of selected miRNAs was determined by in situ hybridization. Functional analysis of miR-30a was performed in zebrafish using antisense-mediated miRNA knockdown. Targets of miR-30a were identified by microarray analysis of gene expression following knockdown in cultured cells. RESULTS: A set of 38 differentially expressed fetal hepatic miRNAs was identified. Several of these miRNAs were found to exhibit distinct temporal and spatial patterns of expression in hepatocytes, cholangiocytes, and nonepithelial cells within the liver. Two (miR-30a and miR-30c) are the first examples of ductal plate and bile duct-specific hepatic miRNAs. Knockdown of miR-30a in the zebrafish larva results in defective biliary morphogenesis. Several newly identified targets of miR-30a are known regulators of liver development and function. CONCLUSIONS: We have identified miRNAs whose spatial and temporal patterns of expression are suggestive of functional roles in hepatic development and/or function. One of these, the biliary miRNA miR-30a, is required for biliary development in zebrafish. This is the first demonstration of a functional role for miRNA in hepatic organogenesis. PMID- 19185581 TI - Glucocorticoids and tumor necrosis factor-alpha synergize to induce absorption by the epithelial sodium channel in the colon. AB - BACKGROUND & AIMS: The epithelial sodium channel (ENaC) mediates electrogenic sodium absorption in distal colon. In patients with inflammatory bowel disease (IBD), ENaC induction is impaired, mainly through transcriptional suppression by proinflammatory cytokines such as tumor necrosis factor (TNF)-alpha. Glucocorticoid therapy rapidly increases sodium absorption; we investigated the molecular mechanisms underlying the interaction among TNF-alpha, glucocorticoids, and ENaC induction. METHODS: ENaC-mediated sodium transport in glucocorticoid receptor (GR)-expressing HT-29/B6 cells and rat distal colon, under the influence of the synthetic glucocorticoid dexamethasone and TNF-alpha, was quantified in Ussing chambers. ENaC messenger RNA (mRNA) levels were monitored by real-time polymerase chain reaction. GR transactivation and expression were investigated by gene reporter, immunoblot, and confocal immunofluorescence microscopy analyses. The GR mRNA half-life was determined. Signaling pathways were characterized using mitogen-activated protein kinase inhibitors. RESULTS: Dexamethasone not only prevented TNF-alpha-mediated ENaC suppression but caused synergistic induction of ENaC-dependent sodium absorption in HT-29/B6-GR cells and rat distal colon. This synergy resulted from TNF-alpha-mediated increases in GR protein levels because of GR mRNA stabilization and subsequent GR transactivation by dexamethasone. As a consequence, transcription of the ENaC beta- and gamma-subunits was up-regulated, increasing ENaC-dependent sodium absorption. p38 Mitogen-activated protein kinase is required for this synergistic effect: p38 inhibition blocked the increase in GR protein expression and ENaC-dependent sodium absorption. CONCLUSIONS: TNF alpha and dexamethasone induce ENaC, explaining the rapid and intense proabsorptive effect of glucocorticoid therapies. PMID- 19185582 TI - Tissue-specific amino acid transporter partners ACE2 and collectrin differentially interact with hartnup mutations. AB - BACKGROUND & AIMS: Hartnup amino acid transporter B(0)AT1 (SLC6A19) is the major luminal sodium-dependent neutral amino acid transporter of small intestine and kidney proximal tubule. The expression of B(0)AT1 in kidney was recently shown to depend on its association with collectrin (Tmem27), a protein homologous to the membrane-anchoring domain of angiotensin-converting enzyme (ACE) 2. METHODS: Because collectrin is almost absent from small intestine, we tested the hypothesis that it is ACE2 that interacts with B(0)AT1 in enterocytes. Furthermore, because B(0)AT1 expression depends on an associated protein, we tested the hypothesis that Hartnup-causing B(0)AT1 mutations differentially impact on B(0)AT1 interaction with intestinal and kidney accessory proteins. RESULTS: Immunofluorescence, coimmunoprecipitation, and functional experiments using wild-type and ace2-null mice showed that expression of B(0)AT1 in small intestine critically depends on ACE2. Coexpressing new and previously identified Hartnup disorder-causing missense mutations of B(0)AT1 with either collectrin or ACE2 in Xenopus laevis oocytes showed that the high-frequency D173N and the newly identified P265L mutant B(0)AT1 transporters can still be activated by ACE2 but not collectrin coexpression. In contrast, the human A69T and R240Q B(0)AT1 mutants cannot be activated by either of the associated proteins, although they function as wild-type B(0)AT1 when expressed alone. CONCLUSIONS: We thus show that ACE2 is necessary for the expression of the Hartnup transporter in intestine and suggest that the differential functional association of mutant B(0)AT1 transporters with ACE2 and collectrin in intestine and kidney, respectively, participates in the phenotypic heterogeneity of human Hartnup disorder. PMID- 19185583 TI - A population-based study of perforated diverticular disease incidence and associated mortality. AB - BACKGROUND & AIMS: Perforated diverticular disease represents the most serious complication of diverticular disease, but little is known regarding its occurrence and mortality. We aimed to determine the incidence and mortality associated with diverticular perforation and the influence of comorbidity. METHODS: We used a population-based cohort study using patients with perforated diverticular disease and population controls identified from 1990 to 2005 in the General Practice Research Database (GPRD). Incidence and mortality rates were modelled using Poisson and Cox regression. Comorbidity was quantified using the Charlson index. RESULTS: We identified 953 incident patients. The overall incidence was 2.66 (95% confidence interval [CI]: 2.49-2.83) per 100,000 person years. The incidence rates increased 2.28-fold (95% CI: 1.79-2.95) when corrected for age and sex between 1990 and 2005. The risk of death was highest in the first year with a 6-fold increase (hazard ratio [HR], 5.63; 95% CI: 4.68-6.77). Adjusted for age and sex, the risk of death in the first year was highest in those with lowest comorbidity (HR, 11.11; 95% CI: 8.06-15.31), but the absolute mortality rates were greatest in those with the highest comorbidity (263.1 per 1000 person-years). CONCLUSIONS: The incidence of perforated diverticular disease has doubled over the period of the study. Patients presenting with a perforated diverticulum are 6 times more likely to die than the general population during the first year following perforation. Those who have the greatest comorbidity are the most likely to die; however, those with least comorbidity have an 11-fold increase in mortality in the first year. PMID- 19185584 TI - Influence of methoprene and dietary protein on male Anastrepha suspensa (Diptera:Tephritidae) mating aggregations. AB - The Caribbean fruit fly, Anastrepha suspensa (Loew), like many polyphagous tephritids, exhibits a lek polygyny mating system, and juvenile hormone levels and adult diet are known to have important positive effects on male sexual success. Among the potential components of this success are male lek tenure and female response to the sexual signals of lekking males. Male A. suspensa where submitted to four different treatments: (M(+)P(+)) application of juvenile hormone analog, methoprene (M) and sugar and hydrolyzed yeast as adult food; (M(+)P(-)) application of M and sugar as adult food; (M(-)P(+)) no application of M and sugar and hydrolyzed yeast as adult food; and (M(-)P(-)) no application of M and sugar as adult food. M(+)P(+) males initiated and participated more in aggregations, mated more frequently, and occupied the lek centers more often. They also had fewer unsuccessful mounting attempts than males in all the other treatments. M(+)P(+) males also emitted pheromones and acoustically signaled more often and attracted more females than males in other treatments. Male sexual performance was improved due to methoprene, protein supply, and the interaction of methoprene and protein for most of the parameters. Since the success of the sterile insect technique (SIT), a commonly employed technique to control pest tephritids, requires the release of males that can form leks, engage in agonistic sexual interactions, and attract females, these positive effects of protein and methoprene may substantially improve SIT programs. PMID- 19185585 TI - Logical analysis of the budding yeast cell cycle. AB - The budding yeast Saccharomyces cerevisiae is a model organism that is commonly used to investigate control of the eukaryotic cell cycle. Moreover, because of the extensive experimental data on wild type and mutant phenotypes, it is also particularly suitable for mathematical modelling and analysis. Here, I present a new Boolean model of the budding yeast cell cycle. This model is consistent with a wide range of wild type and mutant phenotypes and shows remarkable robustness against perturbations, both to reaction times and the states of component genes/proteins. Because of its simple logical nature, the model is suitable for sub-network analysis, which can be used to identify a four node core regulatory circuit underlying cell cycle regulation. Sub-network analysis can also be used to identify key sub-dynamics that are essential for viable cell cycle control, as well as identifying the sub-dynamics that are most variable between different mutants. PMID- 19185586 TI - The role of weak selection and high mutation rates in nearly neutral evolution. AB - Neutral dynamics occur in evolution if all types are 'effectively equal' in their reproductive success, where the definition of 'effectively equal' depends on the population size and the details of mutations. Empirically observed neutral genetic evolution in extremely large clonal populations can only be explained under current models if selection is completely absent. Such models typically consider the case where population dynamics occurs on a different timescale to evolution. However, this assumption is invalid when mutations are not rare in a whole population. We show that this has important consequences for the occurrence of neutral evolution in clonal populations. In highly connected type spaces, neutral dynamics can occur for all population sizes despite significant selective differences, via the forming of effectively neutral networks connecting rare neutral types. Biological implications include an explanation for the high diversity of rare types that survive in large clonal populations, and a theoretical justification for the use of neutral null models. PMID- 19185587 TI - Characterization of a C-type lectin from the cotton bollworm, Helicoverpa armigera. AB - C-type lectins can specifically recognize sugars on the surface of microorganisms and cause a series of immune responses to effectively resist pathogenic invasions. In previous work in our laboratory, we obtained a C-type lectin from Helicoverpa armigera (Ha-lectin). It has two different carbohydrate recognition domains (CRDs) CRD1 and CRD2 arranged in tandem. In this study, recombinant CRD1 and CRD2 were expressed separately in Escherichia coli and purified. They have the ability to agglutinate Gram-negative and Gram-positive bacteria and fungi in the presence of Ca2+. They also have different spectra of sugar binding abilities. The rHa-lectin, rCRD1 and rCRD2 could inhibit the growth in quantity of Bacillus thuringiensis in vivo by increasing hemocyte phagocytosis. These results suggested that Ha-lectin and its two domains could function as a pattern recognition receptor or an opsonin in vivo to promote the hemocyte phagocytosis of pathogens and protect the insect from bacterial infection. PMID- 19185588 TI - Cflec-4, a multidomain C-type lectin involved in immune defense of Zhikong scallop Chlamys farreri. AB - C-type lectins are a superfamily of carbohydrate-recognition proteins which play crucial roles in the innate immunity. In this study, a novel multidomain C-type lectin gene from scallop Chlamys farreri (designated as Cflec-4) was cloned by RACE approach based on EST analysis. The full-length cDNA of Cflec-4 was of 2086 bp. The open reading frame was of 1830bp and encoded a polypeptide of 609 amino acids, including a signal sequence and four dissimilar carbohydrate-recognition domains (CRDs). The deduced amino acid sequence of Cflec-4 shared high similarities to other C-type lectin family members. The phylogenetic analysis revealed the divergence between the three N-terminal CRDs and the C-terminal one, suggesting that the four CRDs in Cflec-4 originated by repeated duplication of different primordial CRD. The potential tertiary structure of each CRD in Cflec-4 was typical double-loop structure with Ca2+-binding site 2 in the long loop region and two conserved disulfide bridges at the bases of the loops. The tissue distribution of Cflec-4 mRNA was examined by fluorescent quantitative real-time PCR. In the healthy scallops, the Cflec-4 transcripts could be only detected in gonad and hepatopancreas, whereas in the Listonella anguillarum challenged scallops, it could be also detected in hemocytes. These results collectively suggested that Cflec-4 was involved in the immune defense of scallop against pathogen infection and provided new insight into the evolution of C-type lectin superfamily. PMID- 19185589 TI - Modulation of TNFalpha, IL-10 and IL-12p40 levels by a ceramide-1-phosphate analog, PCERA-1, in vivo and ex vivo in primary macrophages. AB - Phospho-ceramide analog-1 (PCERA-1) has been described as a potent in vivo suppressor of the pro-inflammatory cytokine tumor necrosis factor alpha (TNFalpha), and thus as a putative drug for the treatment of inflammatory diseases. However, the in vivo cell target of PCERA-1 has not been identified, and its in vivo effect on secretion of other relevant cytokines has not been reported. We have previously shown that PCERA-1 suppresses lipopolysaccharide (LPS)-induced TNFalpha production in RAW264.7 macrophages in vitro. We therefore hypothesized that PCERA-1 targets TNFalpha production by primary macrophages. In this study we thus investigated the effect of PCERA-1 on LPS-induced release of TNFalpha, interleukin (IL)-10 and IL-12p40, in vivo, and ex vivo. We found that PCERA-1 suppressed production of the pro-inflammatory cytokines, TNFalpha and IL 12p40, and increased production of the anti-inflammatory cytokine, IL-10, in LPS challenged mice, and in primary peritoneal macrophages as well as bone marrow derived macrophages (BMDM) stimulated with LPS and interferon (IFN)-gamma. These activities of PCERA-1 were independent of each other. In contrast, PCREA-1 only slightly affected TNFalpha production in the whole blood assay, where LPS-induced cytokines are mainly produced by monocytes. Moreover, isolated blood monocytes were inert to PCERA-1, but acquired responsiveness to PCERA-1 upon macrophage colony stimulating factor (M-CSF)-induced differentiation into macrophages. Pharmacokinetic analysis in mice showed that while the volume of distribution of PCERA-1 is low, the drug was rapidly exchanged between the peritoneum and the systemic circulation. Together, these results suggest that sensitivity to PCERA-1 increases upon differentiation of blood monocytes into tissue macrophages, and imply a mechanistic role for peritoneal macrophages in the in vivo anti inflammatory activity of PCERA-1. Finally, we show that the mechanism of activity of PCERA-1 and prostaglandin E2 (PGE2) is distinct, and that PCERA-1 signaling is not mediated by EP2, a PGE2 receptor which is also activated by oxidized phospholipids. The independent and reciprocal modulation of production of TNFalpha and IL-12p40, vs. IL-10, suggests that PCERA-1 may be a candidate drug for the treatment of inflammation-linked diseases. PMID- 19185590 TI - The polo-like kinase 1 regulates CDC25B-dependent mitosis entry. AB - Activation of cyclin-dependent kinase complexes (CDK) at key cell cycle transitions is dependent on their dephosphorylation by CDC25 dual-specificity phosphatases (CDC25A, B and C in human). The CDC25B phosphatase plays an essential role in controlling the activity of CDK1-cyclin B complexes at the entry into mitosis and together with polo-like kinase 1 (PLK1) in regulating the resumption of cell cycle progression after DNA damage-dependent checkpoint arrest in G2. In this study, we analysed the regulation of CDC25B-dependent mitosis entry by PLK1. We demonstrate that PLK1 activity is essential for the relocation of CDC25B from the cytoplasm to the nucleus. By gain and loss of function analyses, we show that PLK1 stimulates CDC25B-induced mitotic entry in both normal conditions and after DNA-damage induced G2/M arrest. Our results support a model in which the relocalisation of CDC25B to the nucleus at the G2-M transition by PLK1 regulates its mitotic inducing activity. PMID- 19185591 TI - Identification, characterization and essentiality of the unusual peroxin 13 from Trypanosoma brucei. AB - Peroxin 13 (PEX13) is one of the components of a peroxisomal membrane complex involved in import of proteins into the matrix of the organelles and has previously been characterized in a variety of organisms. Trypanosomatids (Trypanosoma, Leishmania), protozoan parasites having peroxisome-like organelles designated glycosomes, possess an unusual PEX13 which shares very low sequence identity with others and lacks some typical PEX13 characteristics. It was identified in the databases through its multiple YGx motifs present in a glycine rich N-terminal region of low sequence complexity. Like other PEX13s, it contains predicted transmembrane segments and a SH3 domain in its C-terminal half. The localization of T. brucei PEX13 in the glycosomal membrane was confirmed by expression of a fusion construct with Green Fluorescent Protein, and western blot analysis of purified organelles and membranes. The C-terminal half of the protein was shown to interact with the third of three pentapeptide repeats of the previously characterized PEX5, the receptor of glycosomal proteins with a type 1 peroxisome-targeting signal, and with PEX14, another component of the same peroxisomal protein import complex in the membrane. PEX13 is essential for the parasite; depletion by RNA interference results in mislocalization of glycosomal proteins and death of the parasites. PMID- 19185592 TI - Identification of a virus-specific and conserved B-cell epitope on NS1 protein of Japanese encephalitis virus. AB - NS1 protein of Japanese encephalitis virus (JEV) is an important non-structural protein, which is able to induce protective immune response in target animals and can be used as specific serological diagnosis tool, but the epitopes on NS1 of JEV have not been identified. For epitope mapping, in this study, a series of 51 partially overlapping fragments covering entire NS1 protein were expressed with a GST-tag and then screened by a monoclonal antibody (mAb). Through enzyme-linked immunosorbent assay (ELISA), linear epitope-containing fragment, the overlapping region of NS1-18 and NS1-19 (residues 145-152), was located. Then a set of peptides derived from that overlapping region with deletions were expressed and subjected to ELISA and Western blot for further mapping purpose. Results indicated that the motif of (146)EHARW(150) is the minimal unit of the linear epitope recognized by that monoclonal antibody (mAb). Western blot showed that this epitope could be recognized by JEV-positive serum from pigs. Furthermore, it was found that the epitope is highly conserved among JEV strains through sequence alignments analysis. Notably, none of the homologous regions on NS1 proteins of other flavivirus could react with the mAb when they were tested for cross reactivity, suggesting the potential clinical application of this epitope in differential diagnosis. PMID- 19185593 TI - Complete sequence of the genome of avian paramyxovirus type 9 and comparison with other paramyxoviruses. AB - The complete genome consensus sequence was determined for avian paramyxovirus (APMV) serotype 9 prototype strain PMV-9/domestic Duck/New York/22/78. The genome is 15,438 nucleotides (nt) long and encodes six non-overlapping genes in the order of 3'-N-P/V/W-M-F-HN-L-5' with intergenic regions of 0-30 nt. The genome length follows the "rule of six" and contains a 55-nt leader sequence at the 3' end and a 47-nt trailer sequence at the 5' end. The cleavage site of the F protein is I-R-E-G-R-I downward arrowF, which does not conform to the conventional cleavage site of the ubiquitous cellular protease furin. The virus required exogenous protease for in vitro replication and grew only in a few established cell lines, indicating a restricted host range. Alignment and phylogenetic analysis of the predicted amino acid sequences of APMV-9 proteins with the cognate proteins of viruses of all five genera of family Paramyxoviridae showed that APMV-9 is more closely related to APMV-1 than to other APMVs. The mean death time in embryonated chicken eggs was found to be more than 120h, indicating APMV-9 to be avirulent for chickens. PMID- 19185594 TI - The flaviviral methyltransferase is a substrate of Casein Kinase 1. AB - Serine/Threonine phosphorylation of the nonstructural protein 5 (NS5) is a conserved feature of flaviviruses, but the identity and function(s) of the responsible kinase(s) remain unknown. Serine 56 in the methyltransferase domain of NS5 can be phosphorylated intracellularly, is conserved in all flaviviruses, and is a critical residue in the catalytic mechanism. A negative charge at this residue inactivates the 2'-0 methyltransferase activity necessary to form a 5' cap structure of the viral RNA. Here we show pharmacologic inhibition of Casein Kinase 1 (CK1) suppresses yellow fever virus (YFV) production. We also demonstrate the alpha isoform of Casein Kinase 1 (CK1alpha), a kinase previously identified as phosphorylating Hepatitis C Virus NS5A protein, also phosphorylates serine 56 of YFV methyltransferase. Overall these results suggest CK1 activity can influence flaviviral replication. PMID- 19185595 TI - Characterization of classical swine fever virus (CSFV) nonstructural protein 3 (NS3) helicase activity and its modulation by CSFV RNA-dependent RNA polymerase. AB - Classical swine fever virus (CSFV) nonstructural protein 3 (NS3) is believed to possess three enzyme activities that are likely to be essential for virus replication: a serine protease located in the N-terminus and NTPase as well as helicase activities located in the C-terminus. In this report, we expressed NS3 helicase domain (NS3h) in E. coli and characterized its helicase activity. The NS3h helicase activity was dependent on the presence of NTP and divalent cations, with a preference for ATP and Mn(2+), and required the substrates possessing a 3' un-base-paired region on the RNA template strand. The NS3h helicase activity was proportional to increasing lengths of the 3' un-base-paired regions up to 16 nucleotides of the RNA substrates. We also investigated the modulation of NS3 NTPase/helicase activities by NS3 protease domain and NS5B, an RNA-dependent RNA polymerase (RdRp). Our data showed that the NS3 protease domain enhanced the helicase activity of NS3 but had no effect on its NTPase activity. For the truncated NS3 (helicase domain, NS3h), both NTPase and helicase activities were up-regulated by NS5B. However, for the full-length NS3 (NS3FL), the NTPase activity, but not the helicase activity, was stimulated by NS5B. Maltose-binding protein (MBP) pull-down as well as enzyme-linked immunosorbent assays confirmed the specific interaction between NS3 and NS5B. PMID- 19185596 TI - SARS coronavirus spike protein-induced innate immune response occurs via activation of the NF-kappaB pathway in human monocyte macrophages in vitro. AB - A purified recombinant spike (S) protein was studied for its effect on stimulating human peripheral blood monocyte macrophages (PBMC). We examined inflammatory gene mRNA abundances found in S protein-treated PBMC using gene arrays. We identified differential mRNA abundances of genes with functional properties associated with antiviral (CXCL10) and inflammatory (IL-6 and IL-8) responses. We confirmed cytokine mRNA increases by real-time quantitative(q) RT PCR or ELISA. We further analyzed the sensitivity and specificity of the prominent IL-8 response. By real-time qRT-PCR, S protein was shown to stimulate IL-8 mRNA accumulation in a dose dependent manner while treatment with E protein did not. Also, titration of S protein-specific production and secretion of IL-8 by ELISA showed that the dose of 5.6nM of S produced a significant increase in IL 8 (p=0.003) compared to mock-treated controls. The increase in IL-8 stimulated by a concentration of 5.6nM of S was comparable to concentrations seen for S protein binding to ACE2 or to neutralizing monoclonal antibody suggesting a physiological relevance. An NF-kappaB inhibitor, TPCK (N-Tosyl-L-Phenylalanine Chloromethyl Ketone) could suppress IL-8 production and secretion in response to S protein in PBMC and THP-1 cells and in HCoV-229E virus-infected PBMC. Activation and translocation of NF-kappaB was shown to occur rapidly following exposure of PBMC or THP-1 cells to S protein using a highly sensitive assay for active nuclear NF kappaB p65 transcription factor. The results further suggested that released or secreted S protein could activate blood monocytes through recognition by toll like receptor (TLR)2 ligand. PMID- 19185597 TI - Modeling gene sequence changes over time in type 3 dengue viruses from Ecuador. AB - Dengue virus (DENV) is a member of the genus Flavivirus of the family Flaviviridae. DENV-3 re-emerged in Central America in 1994, and continues to expand into the South American region. Little is known about the evolutionary rates, viral spread and population dynamics of this genotype in the Latin American region. In order to gain insight into these matters, we used a Bayesian Markov chain Monte Carlo (MCMC) approach, to analyze envelope (E) gene sequences of the DENV-3 genotype III of strains included in a monophyletic cluster composed by Ecuadorian as well as strains from Cuba, Puerto Rico and Peru. The results of these studies revealed that the expansion population growth model was the best fit to the data. The most common recent ancestor (MRCA) was placed around 1989, in agreement with the first reports of the emergence of this new DENV-3 type. A mean rate 1.033 x 10(-3) nucleotide substitution per site per year was obtained. This rate is comparatively higher than the ones obtained for DENV-2 and DENV-4 in the same region. Faster population growth and greater population dispersal may have contributed to the vigorous initial transmission dynamics of this genotype in the Latin American region. PMID- 19185598 TI - Identification of the nonstructural protein 4B of hepatitis C virus as a factor that inhibits the antiviral activity of interferon-alpha. AB - Interferon-alpha (IFN-alpha) is the most commonly used therapeutics for the treatment of chronic viral infection. However, many viruses are resistant to IFN alpha treatment to some degrees through encoding inhibitors of the IFN-alpha producing or signaling pathway. Multiple HCV viral proteins have been reported to be involved in IFN-alpha resistance. To develop a method to screen for factors that inhibit the antiviral activity of IFN-alpha, a mini-library of HCV genome was transduced into the Huh7 cells containing the HCV subgenomic replicon (CON1 HCV S2204I) and screened for the factor that rendered the cells more resistant to IFN-alpha treatment. A fragment of nonstructural protein 4B (NS4B), named tNS4B, was isolated. Expression of tNS4B or the full-length NS4B in CON1 HCV S2204I or naive Huh7 cells inhibited the protection of the cells by IFN-alpha treatment from vesicular stomatitis virus (VSV) infection. In Huh7 cells expressing NS4B or tNS4B, IFN-alpha-induced phosphorylation levels of signal transducer and activator of transcription 1 (STAT1) were reduced. Furthermore, expression of NS4B reduced IFN-alpha-induced expression levels of type I interferon receptor and a reporter driven by the ISRE promoter. In conclusion, we have developed a method to screen for IFN-alpha resistance factors and identified HCV NS4B as such a factor. PMID- 19185599 TI - Molecular characterization of Rabies Virus isolates from dogs and crab-eating foxes in Northeastern Brazil. AB - Thirty-eight samples of Rabies Virus isolated from dogs and crab-eating foxes (Cerdocyon thous) in Northeastern Brazil were characterized genetically by analyzing the G gene and the psi region. The results show that there are two groups of Rabies Virus lineages circulating among domestic and wild animals in the region. The topologies of the phylogenetic trees of the G gene and psi region are similar and reveal the existence of geographic groups. The genetic diversity of the lineages isolated from wild animals (wild group) was approximately twice that of the lineages isolated from domestic animals (domestic group), and the genetic distance between the two groups was 9.93%. Polymorphism analysis revealed specific intra- and inter-group molecular signatures for both the G gene and psi region. Together with the analysis of the N gene undertaken previously, the results of this study confirm the existence of a Rabies Virus phylogroup in Northeastern Brazil (NB) circulating in the C. thous population, making this species a rabies biotype in the region. PMID- 19185600 TI - Parainfluenza virus type 5 (PIV-5) morphology revealed by cryo-electron microscopy. AB - The knowledge of parainfluenza type 5 (PIV-5) virion morphology is essentially based on the observation of negatively stained preparations in conventional transmission electron microscopy (CTEM). In this study, the ultrastructure of frozen-hydrated intact PIV-5 was examined by cryo-electron microscopy (cryo-EM). Cryo-EM revealed a majority of spherical virions (70%), with a lower pleiomorphy than originally observed in CTEM. Phospholipid bilayer thickness, spike length and glycoprotein spikes density were measured. About 2000 glycoprotein spikes were present in an average-sized spherical virion. Altogether, these data depict a more precise view of PIV-5 morphology. PMID- 19185601 TI - State-dependence of climbing fiber-driven calcium transients in Purkinje cells. AB - The recently described bi-stability of Purkinje cells and the state-dependence of the complex spike waveform suggest that calcium currents may play a pivotal role in both the complex spike waveform and the state of the membrane voltage. Here we used Ca2+ imaging to record the changes in intracellular [Ca2+] that are elicited by either spontaneous or climbing fiber-evoked activity in rat Purkinje cells. We show that a continuous somatic Ca2+ influx occurs during an "UP" state. Furthermore Ca2+ transients that are evoked by climbing fiber stimulation are state-dependent. Somatic transients are smaller following an "UP" state, while dendritic transients are smaller following a "DOWN" state. The state-dependence of these signals should affect the intrinsic firing of Purkinje cells as well as plastic processes that modulate synaptic strength. PMID- 19185602 TI - Hippocampal infusions of apolipoprotein E peptides induce long-lasting cognitive impairment. AB - The inheritance of the varepsilon4 allele of apolipoprotein E (ApoE4) and cholinergic system dysfunction have long been associated with the pathology of Alzheimer's disease (AD). Recently, in vitro studies have established a direct link between ApoE and cholinergic function in that synthetic peptides containing segments of the ApoE protein (ApoE(133-149) and ApoE(141-148)) interact with alpha7 nicotinic acetylcholine receptors (nAChRs) in the hippocampus. This raises the possibility that ApoE peptides may contribute to cognitive impairment in AD in that the hippocampus plays a key role in cognitive functioning. To test this, we acutely infused ApoE peptides into the ventral hippocampus of female Sprague Dawley rats and assessed the resultant effects on radial-arm maze choice accuracy over a period of weeks after the infusion. Local ventral hippocampal infusion of ApoE peptides caused significant cognitive impairment in radial-arm maze learning that persisted several weeks after the acute infusion. This persisting deficit may be an important model for understanding the relationship between ApoE protein induced neurotoxicity and cognitive impairment as well as serve as a platform for the development of new therapies to avoid neurotoxicity and cognitive decline. PMID- 19185603 TI - MCP-1 involvement in glial differentiation of neuroprogenitor cells through APP signaling. AB - Previously it has been reported that neural stem cells undergoing apoptotic stress have increased levels of amyloid precursor protein (APP) and increased APP expression results in glial differentiation. APP activity was also shown to be required for staurosporine-induced glial differentiation of neuroprogenitor cells. Monocyte chemoattractant protein-1 (MCP-1) is a chemokine that is expressed early during inflammation. The binding of MCP-1 to its chemokine receptor induces expression of novel transcription factor MCP-1-induced protein (MCPIP). MCPIP expression subsequently leads to cell death. Previous studies have shown that pro-apoptotic factors have the ability to induce neural differentiation. Therefore, we investigated if MCPIP expression leads to differentiation of NT2 neuroprogenitor cells. Results showed that MCPIP expression increased glial fibrillary acid protein (GFAP) expression and also caused distinct morphological changes, both indicative of glial differentiation. Similar results were observed with MCP-1 treatment. Interestingly, APP expression decreased in response to MCPIP. Instead, we found APP activity regulates expression of both MCP-1 and MCPIP. Furthermore, inhibition of either p38 MAPK or JAK signaling pathways significantly reduced APP's effect on MCP-1 and MCPIP. These data demonstrates the role APP has in glial differentiation of NT2 cells through MCP-1/MCPIP signaling. It is possible that increased APP expression after CNS injury could play a role in MCP-1 production, possibly promoting astrocyte activation at injured site. PMID- 19185604 TI - MRI protocol for in vivo visualization of the Gottingen minipig brain improves targeting in experimental functional neurosurgery. AB - BACKGROUND: The Gottingen minipig is increasingly used as an animal model in experimental neuroscience as a much needed alternative to non-human primates. Accurate spatial targeting in this species in vivo is challenging, and most clinically available magnetic resonance imaging (MRI) protocols do not provide sufficient spatial resolution for this purpose. Thus, the aim of this study was to develop an in vivo pre-operative MRI protocol allowing direct visualization of individual nuclei of major interest in the minipig brain. MATERIALS AND METHODS: Three Gottingen minipigs underwent MRI using an inversion-recovery fast spin-echo sequence that was optimized with regards to the following parameters: inversion time, relaxation time, echo time and spatial and temporal resolution, giving a scan duration acceptable for the tight schedule usually employed in a neurosurgical procedure. The most optimal pulse sequence was applied in 8 Gottingen minipigs and the anatomical structures were identified. RESULTS AND CONCLUSION: High-resolution images with excellent contrast were acquired, presenting negligible geometric distortions. Minor flow artifacts from the large neck vessels generated the most prominent artifact. Determination of coordinates necessary in experimental neurosurgery in the Gottingen minipig was considerably improved with this MRI protocol. PMID- 19185605 TI - The effects of electrical stimulation or an electrolytic lesion in the mediodorsal thalamus of the rat on survival, body weight, food intake and running activity in the activity-based anorexia model. AB - The glucose metabolism in the mediodorsal thalamus (MD) is increased in rats in the activity-based anorexia (ABA) model. In patients, electrical stimulation in hyperactive brain regions reduced symptoms in e.g. major depressive disorder and cluster headache. In two blinded randomised controlled experiments, we therefore examined the effects of high-frequency electrical stimulation and an electrolytic lesion in the MD in a validated rat model for anorexia nervosa. The ABA model was successfully replicated in all our experiments, with a reduction in body weight, food intake, and survival time and an increase in running activity. In a first experiment, we evaluated the effect of electrical stimulation or a curative lesion in the MD on survival, body weight, food intake and locomotor activity in ABA rats. Electrical MD stimulation or an electrolytic MD lesion did not improve the symptoms of rats in the ABA model, compared to control groups. In a second experiment, we investigated the effect of a preventive electrolytic lesion in the MD on rats in the ABA model. Although there was no significant improvement of survival, body weight and food intake, locomotor activity was significantly reduced in the lesion group compared to the control group. Apart from this positive effect on running activity, we found no convincing evidence for the suitability of the MD as a neuromodulation target for anorexia nervosa patients. PMID- 19185606 TI - Notch signaling and Hes labeling in the normal and drug-damaged organ of Corti. AB - During the development of the inner ear, the Notch cell signaling pathway is responsible for the specification of the pro-sensory domain and influences cell fate decisions. It is assumed that Notch signaling ends during maturity and cannot be reinitiated to alter the fate of new or existing cells in the organ of Corti. This is in contrast to non-mammalian species which reinitiate Delta 1 Notch1 signaling in response to trauma in the auditory epithelium, resulting in hair cell regeneration through transdifferentiation and/or mitosis. We report immunohistochemical data and Western protein analysis showing that in the aminoglycoside-damaged guinea pig organ of Corti, there is an increase in proteins involved in Notch activation occurring within 24h of a chemical hair cell lesion. The signaling response is characterized by the increased presence of Jagged1 ligand in pillar and Deiters cells, Notch1 signal in surviving supporting cell nuclei, and the absence of Jagged2 and Delta-like1. The pro-sensory bHLH protein Atoh1 was absent at all time points following an ototoxic lesion, while the repressor bHLH transcription factors Hes1 and Hes5 were detected in surviving supporting cell nuclei in the former inner and outer hair cell areas, respectively. Notch pathway proteins peaked at 2 weeks, decreased at 1 month, and nearly disappeared by 2 months. These results indicate that the mammalian auditory epithelium retains the ability to regulate Notch signaling and Notch dependent Hes activity in response to cellular trauma and that the signaling is transient. Additionally, since Hes activity antagonizes the transcription of pro sensory Atoh1, the presence of Hes after a lesion may prohibit the occurrence of transdifferentiation in the surviving supporting cells. PMID- 19185607 TI - Brief heat treatment increases cytotoxicity of Mannheimia haemolytica leukotoxin in an LFA-1 independent manner. AB - Mannheimia haemolytica is an important respiratory pathogen in cattle. Its predominant virulence factor is a leukotoxin (LKT) that is a member of the RTX family of exotoxins produced by a variety of Gram negative bacteria. LKT binds to the CD18 chain of beta(2) integrins on bovine leukocytes, resulting in cell death. In this study, we show that brief heat treatment of native LKT (95 degrees C for 3 min) results in increased cytotoxicity for BL-3 (bovine lymphoblastoid) cells. Similar heat treatment restored the activity of LKT that had been rendered inactive by incubation at 22 degrees C for 3 days. A hallmark of LKT is that its toxicity is restricted to leukocytes from cattle or other ruminant species. Surprisingly, heat treatment rendered LKT cytotoxic for human, porcine and canine leukocytes. Membrane binding studies suggested that heat-treated LKT binds to membrane proteins other than LFA-1, and is distributed diffusely along the BL-3 cell membrane. Circular Dichroism spectroscopy studies indicate that heat treatment induced a small change in the secondary structure of the LKT that was not reversed when the LKT was cooled to room temperature. Thus, we speculate that these structural changes might contribute to the altered biological properties of heat-treated LKT. PMID- 19185608 TI - Proteomic analysis of Vibrio vulnificus M2799 grown under iron-repleted and iron depleted conditions. AB - Vibrio vulnificus is an opportunistic marine bacterium that causes a serious, often fatal, infection in human. An important factor that determines the survival of V. vulnificus in the human body is the ability to acquire iron. The differential expression of proteins in whole-cell lysates of V. vulnificus M2799, a clinical isolate, was evaluated under iron-repleted and iron-depleted conditions during the early, mid and late logarithmic growth phases. A total of 32, 53 and 42 iron-regulated spots were detected by two-dimensional differential gel electrophoresis (2D-DIGE) in the early, mid and late logarithmic growth phases, respectively. Of these, 18 (early logarithmic growth phase), 31 (mid logarithmic growth phase) and 26 (late logarithmic growth phase) proteins were subsequently identified by matrix-assisted laser desorption/ionization-time of flight analysis. These proteins were classified into 10 functional categories, including inorganic ion transport and metabolism, carbohydrate transport and metabolism, and amino acid transport and metabolism. Based on this classification, the expression of proteins involved in the iron acquisition system increased from the early to the mid logarithmic growth phases, while that of proteins involved in other metabolic pathways increased from the mid to the late logarithmic growth phases. Furthermore, when the protein expression profile of the wild type bacterium was compared with that of the fur mutant grown under the iron-repleted condition, the expression of 18 proteins was found to be regulated by iron and Fur. PMID- 19185609 TI - Efficient repair of protein radicals by ascorbate. AB - Protein radicals were selectively generated by reaction with azide radicals on Trp and Tyr residues in insulin, beta-lactoglobulin, pepsin, chymotrypsin, and bovine serum albumin at rate constants in the range (2.9-19)x10(8) M(-1) s(-1). Monohydrogen ascorbate reduced tryptophanyl radicals in chymotrypsin and pepsin with rate constants in the narrow range of (1.6-1.8)x10(8) M(-1) s(-1), whereas beta-lactoglobulin tryptophanyl radicals reacted almost 10 times slower. The corresponding values for the protein tyrosyl radicals were about an order of magnitude smaller. Comparison of the rate constants of reactions of free and protein-bound tryptophanyl and tyrosyl radicals showed that, in most cases, the location of the radicals in the protein chain did not constitute a major barrier to the reaction with monohydrogen ascorbate. The results suggest that, under physiological concentrations of dioxygen, monohydrogen ascorbate is likely to be a significant target of protein radicals. It seems likely, therefore, that reaction with protein radicals may be responsible for much of the well-documented loss of ascorbate in living organisms subjected to oxidative stress. PMID- 19185610 TI - Estrogen deficiency results in apoptosis in the frontal cortex of adult female aromatase knockout mice. AB - The aromatase knockout (ArKO) mouse is completely estrogen deficient. We previously detected apoptosis in the hypothalamus of 1 year-old male ArKO mice. This study shows that 12 week-old female ArKO mice display spontaneous apoptosis of pyramidal neurons in the frontal cortex while wild-type (WT) littermates show no signs of apoptosis. Concomitantly, bcl-2 related anti-apoptotic genes are down regulated whereas the pro-apoptotic gene TRADD is up-regulated in the female ArKO frontal cortex. This phenotype can be rescued by 3-week replacement of 17beta estradiol. Furthermore, the apoptosis phenotype is exacerbated in 12-15 month-old female ArKO mice, which have 30% less neurons in the frontal cortex and lower brain weights than WT counterparts. These data show that estrogens are essential for the survival of female cortical neurons even in the absence of pathological conditions or external assaults. Our observations also demonstrate the sexually dimorphic susceptibility of neurons to estrogen deficiency. PMID- 19185611 TI - Propolis and Herba Epimedii extracts enhance the non-specific immune response and disease resistance of Chinese sucker, Myxocyprinus asiaticus. AB - The effect of traditional Chinese medicine (TCM) formulated from propolis and Herba Epimedii extracts at the ratio of 3:1 (w/w) on non-specific immune response of Chinese sucker (Myxocyprinus asiaticus) was investigated. Fish were fed diets containing 0 (control), 0.1%, 0.5% or 1.0% TCM extracts for five weeks. The respiratory burst and phagocytic activities of blood leukocytes, lysozyme and natural haemolytic complement activities in plasma were measured weekly. After five weeks of feeding, fish were infected with Aeromonas hydrophila and mortalities were recorded. Results of this study showed that feeding Chinese sucker with different dosage of TCM extracts stimulated respiratory burst activity, phagocytosis of phagocytic cells in blood and lysozyme activity in plasma. They had no effect on plasma natural haemolytic complement activity. All dosage of treated groups showed reduced mortality following A. hydrophila infection. Feed containing 0.5% TCM extracts was the most effective with the mortality of the fish significantly reduced by 35% compared to the control. The results indicate that propolis and Herba Epimedii extracts in combination enhances the non-specific immune response and disease resistance of Chinese sucker against A. hydrophila. PMID- 19185612 TI - Hippocampus-dependent strategy choice predicts low levels of cell proliferation in the dentate gyrus. AB - Neurogenesis continues to occur throughout life in the mammalian hippocampus. Previous research has suggested that the production of new neurons in the hippocampus during adulthood may be related to hippocampus-dependent learning and memory. However, the exact relationship between adult neurogenesis and learning and memory remains unclear. Here we investigated whether learning strategy selection is related to cell proliferation or to survival of new neurons in the hippocampus of adult male rats. We trained rats on alternating blocks of hippocampus-dependent (hidden platform) and hippocampus-independent (visible platform) versions of the Morris water task with the platform always in the same position. Following training, rats were given a probe session during which the platform was visible and in a novel location. Preferred strategy was determined by observing the initial swim path. Rats were classified as place strategy (hippocampus-dependent) users if they swam to the old platform location. Cue strategy (hippocampus-independent) users were classified as those rats that swam initially to the visible platform. Our results indicate that rats that preferentially used a place strategy had significantly lower cell proliferation than cue strategy users. However, there was no significant difference in cell survival or number of immature neurons between strategy user groups. These results suggest that low levels of cell proliferation in the dentate gyrus may be conducive or coincident with more efficient memory processing in the hippocampus. PMID- 19185614 TI - Cognitive assessment in patients with epilepsy using the Community Screening Interview for Dementia. AB - OBJECTIVE: Epilepsy is the most common neurological disease worldwide. It may be complicated by cognitive impairments with consequent deleterious effects on education, vocation, and social and family life of affected individuals. We assessed the cognitive functions of Nigerian patients with epilepsy using the modified Community Screening Interview for Dementia (CSID) questionnaire. METHODS: Eighty-two subjects (41 patients with epilepsy and 41 normal controls) participated in the study. The CSID was used to assess language, memory, orientation, attention, calculation, and praxis. The SPSS statistical package was used for data analysis. RESULTS: The mean ages (in years) of patients with epilepsy and controls were 28.32+/-9.22 and 25.98+/-7.72, respectively. The patients with epilepsy performed poorly when compared with the controls (P<0.05) in the domains Language, Memory, Attention and Calculation, and Praxis, whereas there was no statistically significant difference between the patients and controls in Orientation scores. Duration of epilepsy and of antiepileptic drug (AED) therapy had a significant negative impact on all domains of the CSID (P<0.05), whereas type of AED used did not. The CSID had a sensitivity of 91.7%. CONCLUSION: This study showed that patients with epilepsy have significant cognitive impairments compared with controls, and duration of epilepsy and AED therapy have deleterious effects on their cognitive performance. In addition, the CSID has acceptable sensitivity and predictive value. It is recommended that cognitive function assessment should be an integral part of routine evaluation of patients with epilepsy. PMID- 19185613 TI - Perirhinal cortex supports acquired fear of auditory objects. AB - Damage to rat perirhinal cortex (PR) profoundly impairs fear conditioning to 22kHz ultrasonic vocalizations (USVs), but has no effect on fear conditioning to continuous tones. The most obvious difference between these two sounds is that continuous tones have no internal temporal structure, whereas USVs consist of strings of discrete calls separated by temporal discontinuities. PR was hypothesized to support the fusion or integration of discontinuous auditory segments into unitary representations or "auditory objects". This transform was suggested to be necessary for normal fear conditioning to occur. These ideas naturally assume that the effect of PR damage on auditory fear conditioning is not peculiar to 22kHz USVs. The present study directly tested these ideas by using a different set of continuous and discontinuous auditory cues. Control and PR-damaged rats were fear conditioned to a 53kHz USV, a 53kHz continuous tone, or a 53kHz discontinuous tone. The continuous and discontinuous tones matched the 53kHz USV in terms of duration, loudness, and principle frequency. The on/off pattern of the discontinuous tone matched the pattern of the individual calls of the 53kHz USV. The on/off pattern of the 50kHz USV was very different from the patterns in the 22kHz USVs that have been comparably examined. Rats with PR damage were profoundly impaired in fear conditioning to both discontinuous cues, but they were unimpaired in conditioning to the continuous cue. The implications of this temporal discontinuity effect are explored in terms of contemporary ideas about PR function. PMID- 19185615 TI - Neuropsychological findings in patients with Unverricht-Lundborg disease. AB - The aims of this study were to clarify if patients with Unverricht-Lundborg disease (ULD) have adequate cognitive functioning and to delineate their neuropsychological profile. We evaluated 20 patients with ULD and 20 healthy, matched controls. Mean age of the patients was 35 years, and mean duration of disease, 22 years. Patients underwent a neuropsychological battery exploring intelligence, executive functions, visuospatial and verbal memory, depression, and anxiety. Eleven of 20 subjects with ULD had mild to moderate cognitive impairment. Compared with controls, patients with ULD had lower scores on all short-term memory and executive function tasks. Linear regression analysis disclosed significant associations between impaired performance on some memory tests and duration of disease and between severity of myoclonus and performance on most executive function tests. In conclusion, most patients with ULD seem to be impaired with respect to cognitive abilities. Longitudinal prospective studies are needed to confirm and further expand our findings. PMID- 19185616 TI - Effects of cadmium on respiratory burst, intracellular Ca2+ and DNA damage in the white shrimp Litopenaeus vannamei. AB - Acute effects of heavy metal ions on shrimp have been an area of intense study worldwide. However, the molecular mechanism by which cadmium-induced injury occurs remains largely unclear, and methods for mitigating toxicity in vivo have rarely been reported. In this study, the changes in respiratory burst and intracellular free calcium in haemocytes of pacific white shrimp, Litopenaeus vannamei, after exposure to Cd(2+) (CdCl(2)) were examined using flow cytometry. Meanwhile, DNA damage and repair in haemocytes and hepatopancreas cells were studied using the comet assay. Respiratory burst generation, intracellular Ca(2+) concentration ([Ca(2+)]i) and DNA damage in haemocytes and hepatopancreas cells all exhibited a dose-dependent increase and a time-dependent change after treatment with Cd(2+) compared with controls. These results indicate that Cd can induce oxidative stress and DNA damage in the shrimp L. vannamei. Moreover, the results also demonstrate that these parameters can be used as sensitive indicators of exposure to this genotoxicant. PMID- 19185617 TI - Cucurbitacin D isolated from Trichosanthes kirilowii induces apoptosis in human hepatocellular carcinoma cells in vitro. AB - The aim of the present study is to examine the effects of the anti-tumor component isolated from Trichosanthes kirilowii on human hepatocellular carcinoma cells. Using Sephadex G-25 column chromatography, Sep-Pak Plus C18 cartridge and high-performance liquid chromatography (HPLC), we isolated the active component from trichosanthes extract. By fast atom bombardment mass spectrometric analysis, the molecular mass of the active fraction was determined, the active components identified, and their mechanisms of action were analyzed by cell growth assay, cell cycle analysis, TUNEL staining and Western blot analysis. We found that the anti-tumor components isolated from the extract of trichosanthes (EOT) are cucurbitacin D and dihydrocucurbitacin D, and suggest that cucurbitacin D induces apoptosis through caspase-3 and phosphorylation of JNK in hepatocellular carcinoma cells. These results suggest that cucurbitacin D isolated from Trichosanthes kirilowii could be a valuable candidate for anti-tumor drug. PMID- 19185618 TI - Improvement of deficient natural killer activity and delayed bactericidal activity by a thiol proteinase inhibitor, E-64-d, in leukocytes from Chediak Higashi syndrome patients in vitro. AB - We previously reported that administration of a potent calpain inhibitor, E-64-d, which protects protein kinase C (PKC) from proteolysis, in a mouse model of Chediak-Higashi syndrome (CHS) (beige mice), decreases its susceptibility to Staphylococcus aureus infection. In the present study, we examined the in vitro effect of E-64-d on both deficient natural killer (NK) and delayed bactericidal activities of leukocytes from six CHS patients. Our results showed that pretreatment of peripheral blood mononuclear cells (PBMCs) obtained from CHS patients with E-64-d (1 microg/ml) significantly enhanced NK activity against K562 cells. The delayed bactericidal activity of polymorphonuclear cells (PMNs) against S. aureus also showed marked improvement. This was recovered to almost normal levels when PMNs were pretreated with E-64-d (1 microg/ml). On the other hand, the same concentration of E-64-d did not affect either the NK or bactericidal activity of normal controls. In addition, we confirmed that following E-64-d treatment, the abnormal down-regulation of PKC activity after concanavalin A (Con A) stimulation was eliminated in PBMCs obtained from CHS patients. To examine whether PKC is involved in the NK cell-mediated cytolysis and bactericidal activity of PMNs, two potent PKC inhibitors, chelerythrin and GO6976, were used. We found that chelerythrin inhibits NK activity of normal PBMCs in a dose-dependent manner, and GO6976 inhibits NK activity at doses that inhibit Ca(2+)-dependent PKC isozymes. These inhibitors also suppressed the bactericidal activity of PMNs against S. aureus. Taken together, our findings suggested that E-64-d improved the compromised NK and bactericidal activity of leukocytes from CHS patients by reversing the down-regulation of PKC activity. PMID- 19185619 TI - Lung adenocarcinoma and antiphospholipid antibodies. AB - Thrombosis is a frequent finding in cancer patients, being referred to as a poor prognostic factor. The mechanisms underlying the thrombophilic state in malignancy are not well elucidated but involve a complex interaction between tumor and host cells as well as the hemostatic system. A number of studies have demonstrated the presence of antiphospholipid antibodies (aPL) in cancer patients, suggesting a potential role in tumor-associated thrombosis. A prospective analysis has been performed in a group of lung adenocarcinoma patients in respect to the presence of aPL and thrombotic manifestations. Lupus anticoagulant (LAC) was identified in 61 out of 105 patients and it correlated highly with thrombosis (22/61, LAC positive vs 2/44, LAC negative RR=7.93; p<0.001). On the other hand, patients that displayed IgM anti-beta2-glycoprotein I (abeta2GPI) (22/80) showed an unexpected decrease in thrombosis risk (2/22, with IgM abeta2GPI vs 18/58, without IgM abeta2GPI RR=0.29; p=0.04). Considerations on the mechanisms that link cancer, thrombosis and aPL are discussed in this article. PMID- 19185620 TI - On coronary analyses. PMID- 19185622 TI - Light-dark patterns of soluble CD40 ligand: clinical implications. PMID- 19185623 TI - Risk factors and stroke risk stratification for atrial fibrillation. PMID- 19185625 TI - Using CHADS2 backwards plus echo criteria to identify stroke patients who have occult intermittent atrial fibrillation. PMID- 19185626 TI - Ultrafiltration for heart failure: how fast should we move? PMID- 19185627 TI - Health status as a risk factor in cardiovascular disease: a systematic review of current evidence. AB - BACKGROUND: Patient-perceived health status is receiving increased recognition as a patient-centered outcome in chronic heart failure (CHF) and coronary artery disease (CAD), but poor health status is also associated with adverse prognosis. In this systematic review, we examined current evidence on the influence of health status on prognosis in CHF and CAD. METHODS: We conducted a search of PubMed using a set of a priori-defined search terms, the Web of Science for newly cited articles, and the reference lists of eligible articles, resulting in 34 articles. RESULTS: Poor physical health status was a significant predictor for adverse health outcomes in patients with CHF and CAD. In CHF, poor physical health status seemed to be a stronger predictor of hospitalization than mortality. Little evidence was found that poor mental health status is associated with adverse prognosis in CHF and CAD. A disease-specific measure was a better predictor in CHF, but not in CAD. The majority of studies adjusted for an objective measure of disease severity. Neither the index event nor time to follow up appeared to influence the predictive value of health status. CONCLUSIONS: Poor physical health status is associated with adverse CAD and CHF prognosis. Heterogeneity across studies makes definitive conclusions difficult as to which components of health status may be detrimental to patients' health, and how health status as a potential risk factor should be assessed, monitored, and intervened upon in clinical practice. PMID- 19185628 TI - Standardizing care for acute decompensated heart failure in a large megatrial: the approach for the Acute Studies of Clinical Effectiveness of Nesiritide in Subjects with Decompensated Heart Failure (ASCEND-HF). AB - Acute decompensated heart failure (ADHF) is a major public health issue and remains a heterogeneous, complex, and difficult condition to manage. Although novel acute therapies are being tested in large randomized clinical trials, opportunities exist to improve the standard of care by ensuring optimal adherence to currently established evidence-based interventions for HF. Consideration of the goals of therapy and practical application of current methods to assess for clinical improvement may lead to improved patient management and possibly improved outcomes for patients with ADHF. This review provides key practical information regarding the current standard of care for patients with ADHF including the goals of therapy, management of acute and chronic medications, and discharge/transition of care planning. PMID- 19185629 TI - Percutaneous transcatheter aortic valve implantation: Evolution of the technology. AB - Aortic stenosis (AS) is currently the most common valvular abnormality in developed countries. The gold standard treatment of severe symptomatic AS is surgical aortic valve replacement. Mechanical valves were initially the prostheses of choice; with improvement of surgical technique and results, and increased durability of bioprosthetic valves, there has been a trend toward increasing use of bioprostheses. Concurrently, percutaneous technology has advanced exponentially, and the first human percutaneous aortic valve replacement (PAVR) became a reality in 2002. Various groups have now reported their early experiences with PAVR using different technologies. This new treatment modality is evolving very rapidly. There are currently 2 devices being used in clinical trials; several more devices have first-in-man results and others in preclinical development. It appears that PAVR will most likely become a viable option for selected patients in the near future. For these technologies to continue to improve and benefit patients, cross-specialty teamwork is vital. Treatment of severe AS will greatly evolve as these new and emerging technologies improve. Enhancement in device designs and increasing operator experience will make this technology safer and allow its application to a wider patient population. PMID- 19185630 TI - Current concepts in the pathogenesis of atrial fibrillation. AB - Current evidence suggests that the pathogenesis of atrial fibrillation (AF) is multifactorial. The observation that AF, once present, alters the electrophysiologic properties of the atrial myocardium causing self-perpetuation of the arrhythmia raised the importance of electrical remodeling in its pathogenesis. Although these changes are potentially reversible, maintenance of AF continues even after electrical remodeling has occurred. Clinical and experimental studies have highlighted the role of a susceptible atrial anatomical substrate with features of myocyte degeneration and interstitial fibrosis in the initiation and maintenance of AF. Finally, the association of increased inflammatory burden with the presence and future development of AF has implicated inflammation in the pathogenesis of the arrhythmia. The purpose of this review is to provide current evidence on the dominant theories on AF pathogenesis, namely, electrical remodeling, structural remodeling, and inflammation; describe the various experimental models and methods used; and identify a cause-effect association, when present. In addition, the interrelation between different mechanisms responsible for AF will be demonstrated, providing further insight into the complex pathophysiology. PMID- 19185631 TI - Protease-activated receptors in cardiovascular health and diseases. AB - The platelet, once thought to be solely involved in clot formation, is now known to be a key mediator in various other processes such as inflammation, thrombosis, and atherosclerosis. Therefore, antiplatelet agents have become paramount in the prevention and management of various cardiovascular diseases. However, the currently most widely used antiplatelet drugs, aspirin and clopidogrel, have been shown to reduce the risk of serious vascular events only by approximately one quarter. Similarly, oral glycoprotein IIb/IIIa antagonists have been associated with excess mortality, thus restricting the use of parental glycoprotein IIb/IIIa antagonists to the treatment of acute clinical conditions. Thus, for the prevention of cardiovascular diseases, there is still a clinical need for antiplatelet drugs with higher antithrombotic efficacy but with safety profiles that allow for a preventive long-term administration. Thrombin signaling through the protease-activated receptors (PARs) has been shown to influence a wide range of physiologic and pathologic responses in cardiovascular systems. Thus, interference with PARs appears to be a promising strategy to develop new antiplatelet agents with higher efficacy. This review focuses on the cardiovascular actions of PARs that play a role in normal cardiovascular physiology and that are likely to contribute to cardiovascular diseases. PMID- 19185632 TI - Current role of emboli protection devices in percutaneous coronary and vascular interventions. AB - After the observation that distal embolization is a frequent complication of coronary and vascular interventions, mechanical emboli protection devices (EPD) have been developed and tested in different vascular territories. The most frequently used device type incorporates a guidewire with a filter that is placed distal to the target lesion, unfolded, and then retrieved at the end of the procedure. Alternative approaches are based on transient flow obstruction using proximal or distal balloon occlusion. The procedure is then performed under flow reversal or flow arrest and the blood column is evacuated prior to restoration of flow. The efficacy of EPD in reducing major adverse cardiac events among patients undergoing percutaneous intervention of aortocoronary bypass grafts has been demonstrated in a randomized trial. In the acute myocardial infarction setting, EPD failed to improve outcomes. Although randomized data in carotid artery stenting are lacking, a broad, but not unanimous, consensus supports the use of these devices. While a small randomized trial showed disappointing results of emboli protection in renal stenting, no data are available for lower extremity interventions. PMID- 19185633 TI - Rationale and design of the Acute Study of Clinical Effectiveness of Nesiritide in Decompensated Heart Failure Trial (ASCEND-HF). AB - BACKGROUND: Acute decompensated heart failure (ADHF) is a major public health burden with significant mortality and morbidity. Nesiritide is a recombinantly produced intravenous formulation of human B-type natriuretic peptide that promotes vasodilation and increases salt and water excretion, which results in reduced cardiac filling pressures. Prior studies have shown that dyspnea is improved in patients with ADHF 3 hours after nesiritide infusion with significant dose-related reductions in cardiac filling pressures and systemic vascular resistance without significant arrhythmias. However, the effect of nesiritide on dyspnea at 6 or 24 hours is unknown, and no clinical outcome trials have been done to provide a reliable estimate of the effect of nesiritide on morbidity and mortality. METHODS: The Acute Study of Clinical Effectiveness of Nesiritide in Decompensated Heart Failure trial (ASCEND-HF) is a phase III study evaluating the efficacy and safety of nesiritide in patients with ADHF. Patients hospitalized for hear failure will be randomly assigned to receive either intravenous nesiritide or matching placebo for 24 hours to 7 days. The 2 coprimary end points are (1) assessment of acute dyspnea at 6 or 24 hours and (2) death or rehospitalization for hear failure within 30 days. A total of 7,000 patients will be enrolled worldwide between 2007 and 2010. CONCLUSIONS: The data from the ASCEND-HF trial will establish whether nesiritide safely improves acute dyspnea as well as morbidity and mortality at 30 days. PMID- 19185634 TI - A multifaceted intervention to improve blood pressure control: The Guideline Adherence for Heart Health (GLAD) study. AB - BACKGROUND: Although high blood pressure is associated with significant morbidity and mortality, the proportion reaching the goal blood pressures as outlined in the Seventh Report of the Joint National Committee on Prevention, Detection, Evaluation, Treatment of High Blood Pressure (JNC 7) is low. We conducted a randomized trial in primary care practices of a multifactorial intervention targeted to improve providers' adherence to hypertension guidelines. METHOD: A total of 61 primary care practices in North Carolina were randomized to receive either a multifactorial intervention (guideline dissemination via a continuing medical education session, academic detailing sessions, audit and feedback on preintervention rates of adherence, and automated blood pressure machines) or an attention control of similar magnitude but targeted at a different guideline. Outcomes were determined through review of patient charts conducted by an independent masked quality assurance organization. RESULTS: We found no difference between the 2 groups in any of the adherence measures including no difference in the percentage of patients at goal (intervention 49.2%, control 50.6%), with undiagnosed hypertension (18.1% vs 13.6%), average systolic (126 vs 125.1 mm Hg), or diastolic blood pressure (73.1 vs 73.4 mm Hg). Similarly, there was no difference in provider adherence to treatment recommendations (use of thiazide-type diuretic as first-line therapy: 32% vs 29.5%; use of 2-drug therapy in stage 2 hypertension: 11.3% vs 10.4%). CONCLUSION: An intensive, multifactorial intervention did not improve adherence to national hypertension guidelines among community-based primary care. Efforts should be focused on other types of interventions to improve rates of control of hypertension. PMID- 19185636 TI - Daily walking performance as an independent predictor of advanced heart failure: Prediction of exercise capacity in chronic heart failure. AB - PURPOSE: The purpose of this study was to use an accelerometer to measure daily walking performance in patients with chronic heart failure (CHF) to investigate if this parameter is a determinant of New York Heart Association class and indicative of maximal and functional exercise capacity. METHODS: Fifty patients with CHF were instructed to wear an accelerometer for 7 consecutive days while going about their daily business. Maximal and functional exercise capacity was assessed by cardiopulmonary (VO(2peak)) and 6-minute walk testing, respectively. RESULTS: Patients in New York Heart Association I, II, and III reached an average total walking time (TWT) of 160.6 +/- 35.8 minutes, 133.9 +/- 59.0 minutes, and 76.1 +/- 22.5 minutes per day of which 19%, 19%, and 9% where spent in the fast walking mode (>83 m/minute), respectively. The TWT correlated strongly with VO(2peak) (r = 0.72; P <.001) and 6-minute walk testing distance (r = 0.68; P <.001). The TWT and time spent in fast walking mode were the strongest determinants in discriminating moderate CHF. CONCLUSION: Daily walking performance is a clear determinant of maximal and functional exercise capacities in patients with CHF. Walking intensity in particular is an independent predictor in discriminating patients with advanced heart failure. Monitoring of daily walking performance might aid in detecting disease progression and improve clinical outcome. PMID- 19185635 TI - Defining an evidence-based cutpoint for medication adherence in heart failure. AB - BACKGROUND: Despite the importance of medication adherence in heart failure, clinically relevant cutpoints for distinguishing the level of adherence associated with outcomes are unknown. OBJECTIVE: The purpose of this study is to determine the cutpoint above which there is a positive relationship between level of medication adherence and event-free survival. METHODS: This was a longitudinal study of 135 patients with heart failure. Medication adherence was measured using a valid and objective measure, the Medication Event Monitoring System. Two indicators of adherence were assessed by the Medication Event Monitoring System (AARDEX, Union City, CA): (1) dose count, percentage of prescribed doses taken, and (2) dose days, percentage of days the correct number of doses was taken. Patients were followed up to 3.5 years to collect data on outcomes. A series of Kaplan-Meier plots with log-rank tests, Cox survival analyses, and receiver operating characteristic curves were assessed comparing event-free survival in patients divided at one-point incremental cutpoints. RESULTS: Event-free survival was significantly better when the prescribed number of doses taken (dose count) or the correct dose (dose day) was > or =88%. This level was confirmed in a Cox regression model controlling for age, gender, ejection fraction, New York Heart Association, comorbidity, angiotensin-converting enzyme inhibitor use, and beta blocker use. Receiver operating characteristic curves showed that adherence rates above 88% produced the optimal combination of sensitivity and specificity with respect to predicting better event-free survival. With 88% as the adherence cutpoint, the hazard ratio for time to first event for the nonadherent group was 2.2 by dose count (P = .021) and 3.2 by dose day (P = .002). CONCLUSION: The results of this study provide clinicians and researchers with an evidence-based recommendation about the level of adherence needed to achieve optimal clinical outcomes. PMID- 19185637 TI - Incremental value of renal function in risk prediction with the Seattle Heart Failure Model. AB - BACKGROUND: Impaired renal function portends poor heart failure (HF) outcomes. The Seattle Heart Failure Score (SHFS), a multimarker risk assessment tool, however does not incorporate renal function. In this study, we assessed the incremental value of renal function over the SHFS in patients with advanced HF on contemporary optimal treatment. METHODS: Blood urea nitrogen (BUN), serum creatinine (sCr), BUN/sCr ratio, and estimated glomerular filtration rate were assessed in survival models with SHFS as the base model among 443 patients with HF (52 +/- 12 years, male 68.5%, white 52.4%, ejection fraction 0.18 +/- 0.08). Incremental value of renal function was assessed by changes in the likelihood ratio chi(2) and the area under the receiver operating characteristic curves for 1-, 2-, and 3-year event prediction. RESULTS: During a median follow-up of 21 months, 108 (24.5%) of 443 patients had an event (death [n = 92], urgent transplantation [n = 13], or ventricular assist device implantation [n = 3]). All renal parameters individually were associated with outcome (BUN, P < .001; sCr, P < .001; BUN/sCr ratio, P = .006; and estimated glomerular filtration rate, P = .006); however, only BUN was an independent predictor of events in multivariable analyses. Addition of BUN improved the predictive ability of SHFS (Deltalikelihood ratio chi(2) 5.03, P = .025); however, the increase in the area under the receiver operating characteristic curve was marginal (year 1, 0.786 to 0.791; year 2, 0.732 to 0.741; year 3, 0.745 to 0.754; all P > .2). CONCLUSION: Among the various renal function parameters, BUN had the strongest association with outcomes in patients with advanced HF. However, the incremental value of renal function over the SHFS for risk determination was marginal. PMID- 19185638 TI - Differences in initial severity of illness between black and white emergency department patients hospitalized with heart failure. AB - BACKGROUND: Black patients hospitalized for heart failure have better reported short-term survival than white patients for unknown reasons. We sought to determine if initial severity of illness differed between black and white emergency department (ED) patients hospitalized for heart failure. METHODS: We analyzed 1,408 black and 7,260 white randomly selected patients in one state hospitalized from an ED during 2003 and 2004 and with a discharge diagnosis of heart failure. We used three validated clinical prediction rules to estimate severity of illness on admission. RESULTS: Black patients were younger than white patients (65.8 +/- 14.8 vs 77.4 +/- 11.5 years, P < .01) and were assigned to lower risk classes by all 3 prediction rules more frequently than white patients (P < .01). The odds ratio (95% CI) for classification of black versus white patients into the lowest risk class within the three rules ranged from 1.16 (1.00 1.33) to 4.30 (3.75-4.94). After adjusting for hospital clustering, the odds ratio (95% CI) for black versus white patient hospital death and complications was 0.75 (0.60-0.95) and, for 30-day death, was 0.34 (0.27-0.48). CONCLUSIONS: Black ED patients hospitalized with heart failure are younger, less severely ill on admission and less likely to experience short-term fatal and nonfatal outcomes than white patients. Our findings suggest a varying opportunity between black and white patients when considering alternative initial treatment strategies and sites of care. PMID- 19185639 TI - Acute coronary angiographic findings in survivors of out-of-hospital cardiac arrest. AB - BACKGROUND: Diagnosis of acute coronary artery disease in survivors of out-of hospital cardiac arrest (OHCA) is difficult. The role of emergency coronary angiography and percutaneous coronary intervention (PCI) in this setting is debated. The objective of this study was to assess the prevalence of coronary lesions on emergency angiography in survivors of OHCA. METHODS: Seventy-two consecutive OHCA survivors underwent systematic emergency coronary angiography. Patients with critical stenoses or occlusion underwent ad hoc PCI. RESULTS: Most (63.9%) OHCA survivors had angiographic coronary artery disease (> or =1 lesion >50%), but only a minority (37.5%) had clinical or angiographic evidence of an acute coronary syndrome due to either an acute occlusion (16.7%) or an irregular lesion suggestive of ruptured plaque or thrombus (25.0%). A final diagnosis of myocardial infarction was assigned in 27 patients (37.5%). Percutaneous coronary intervention was attempted and successful in 33.3% of the total cohort (n = 24). Hospital survival was 48.6%. By multivariable analysis, use of PCI was not an independent correlate of survival. ST-segment elevation on admission was an independent correlate of acute myocardial infarction (odds ratio 64.2, 95% CI 7.6 544.2, P = .0001), with high positive (82.6%) and negative (83.7%) predictive values. CONCLUSIONS: A minority of OHCA patients has angiographic evidence of an acute coronary syndrome and one-third undergo PCI, but PCI is not an independent correlate of survival. The presence of ST elevation on admission was a strong independent correlate of acute myocardial infarction and may be used to triage OHCA patients to emergency angiography with a view to PCI. PMID- 19185640 TI - Rationale and design of the National Registry of Genetically Triggered Thoracic Aortic Aneurysms and Cardiovascular Conditions (GenTAC). AB - BACKGROUND: Although the management of thoracic aortic aneurysms (TAAs) has improved significantly, patients presenting with aortic dissections, rupture, or other acute complications of TAAs continue to have high rates of morbidity and mortality. Accumulating data have indicated that many TAAs are due to underlying gene mutations. A comprehensive approach to the study of TAAs resulting from genetic mutations is needed to translate this information into advances in treatment. OBJECTIVE: The National Registry of Genetically Triggered Thoracic Aortic Aneurysms and Cardiovascular Conditions (GenTAC) was established to provide a biospecimen inventory and bioinformatics infrastructure to enable research to advance the clinical management of genetically triggered TAAs and related complications. METHODS: GenTAC is a longitudinal observational cohort study enrolling patients with conditions related to genetically induced TAAs from 5 regional clinical centers in the United States. RESULTS: More than 700 subjects with associated clinical histories, physical examinations, imaging data, and biospecimens have been enrolled in the Registry to date. Enrollment is expected to continue until September 2010. Total enrollment of nearly 3,000 subjects is expected. No interim analysis has yet been undertaken. CONCLUSIONS: GenTAC has been established to facilitate studies by GenTAC investigators and others that will advance multiple scientific frontiers in thoracic aortic disease. Genotypic, proteomic, clinical, and imaging data will be integrated systematically with outcomes data to determine the optimal clinical management of patients with genetically induced TAAs. PMID- 19185641 TI - Prognostic value of albuminuria on 1-month mortality in acute myocardial infarction. AB - RATIONALE: An increase in albuminuria occurs in the early days after acute myocardial infarction. The aim of this study was to assess the relation between albuminuria and 30-day mortality, as well as its incremental predictive value, on top of established prognostic parameters. METHODS AND RESULTS: Demographic, clinical, and biological characteristics at admission, as well as in-hospital treatments and 1-month survival, were recorded in 1,211 consecutive patients admitted for acute myocardial infarction. Albuminuria was assessed from an 8-hour overnight urine collection within the first 2 days using immunonephelemetry. The population was categorized into 3 groups according to albuminuria levels (<20, 20 200, and >200 microg/min). Among survivors on day 2, 52% (625/1,211) of patients had an albuminuria level <20 microg/min, 39% (477) between 20 and 200 microg/min, and 9% (109) >200 microg/min. High levels of albuminuria were associated with older age, peripheral vessel disease, systolic blood pressure, glucose, creatinine, troponin, B-type natriuretic peptide, and high-sensitivity C reactive protein levels, as well as use of angiography, angiotensin-converting enzyme inhibitors, and beta blockers. At 1 month, there was a significantly higher mortality rate in groups with higher albuminuria. After adjustment for baseline characteristics, patients with albuminuria level of >20 microg/min had a 2.7-fold higher 30-day mortality, and those with >200 microg/min had an almost 4-fold higher 30-day mortality compared to those with albuminuria level of <20 microg/min. The addition of albuminuria information improved the discrimination capacity of the model and the global risk prediction. CONCLUSIONS: Albuminuria level, taken as a quantitative or categorical variable, is an independent and powerful predictor of mortality after acute myocardial infarction. PMID- 19185642 TI - Angiotensin-converting enzyme inhibition and novel cardiovascular risk biomarkers: results from the Trial of Angiotensin Converting Enzyme Inhibition and Novel Cardiovascular Risk Factors (TRAIN) study. AB - BACKGROUND: Beneficial effects of angiotensin-converting enzyme (ACE) inhibitors seem to be mediated by mechanisms that are partly independent of blood pressure lowering. The present study evaluates effects of an ACE inhibitor (ie, fosinopril) intervention on novel cardiovascular risk factors. METHODS: Data are from the Trial of Angiotensin Converting Enzyme Inhibition and Novel Cardiovascular Risk Factors (TRAIN) study, a double-blind, crossover, randomized, placebo-controlled trial enrolling subjects > or =55 years old with high cardiovascular disease risk profile. Biomarkers of hemostasis (ie, plasminogen activator inhibitor 1, D-dimer), inflammation (ie, C-reactive protein, interleukin-6), and endothelial function (ie, endothelin 1, vascular cell adhesion molecule 1) were measured at the baseline, at the midterm, and at end of follow-up (after 1 year) clinic visits. Paired t test analyses (after Sidak's adjustment, P < .009) were performed to compare biomarkers modifications after fosinopril/placebo interventions. RESULTS: Mean age of the sample (n = 290, women 43.4%) was 66.0 years old. No significant differences were reported for C reactive protein, interleukin 6, plasminogen activator inhibitor 1, vascular cell adhesion molecule 1, and endothelin 1 levels in the comparisons between fosinopril and placebo interventions. D-dimer was the only biomarker showing a significant difference between fosinopril intervention (median 0.32 microg/mL, interquartile range 0.22-0.52 microg/mL) and placebo (median 0.29 microg/mL, interquartile range 0.20-0.47 microg/mL, P = .007) when analyses were restricted to participants with higher compliance to treatment and receiving the maximum ACE inhibitor dosage. CONCLUSIONS: Angiotensin-converting enzyme inhibition does not significantly modify major biomarkers of inflammation, hemostasis, and endothelial function. Further studies should confirm the possible effect of ACE inhibitors on the fibrinolysis pathway. PMID- 19185644 TI - Evaluation of left ventricular short- and long-axis function in severe mitral regurgitation using 2-dimensional strain echocardiography. AB - BACKGROUND: Few data exist on the changes in left ventricular (LV) short- and long-axis function and their usefulness as markers of LV contractile function in patients with chronic, severe mitral regurgitation (MR). METHODS: We studied 59 patients who had severe MR with an ejection fraction > or =50% and 34 healthy controls. Speckle tracking imaging was performed to measure peak systolic radial (SR(R)), circumferential (SR(C)), and longitudinal strain rates (SR(L)). In all patients, the peak rate of LV pressure rise (peak dP/dt) was measured using a micromanometer-tipped catheter. The patients were subdivided into patients with preserved (group 1, peak dP/dt > or =1,300 mm Hg/s [n = 30]) and depressed (group 2 [n = 29]) contractile function. RESULTS: SR(L) was significantly depressed in groups 1 and 2 when compared with the control group, but there was no difference between groups 1 and 2. In contrast, SR(R) and SR(C) were depressed only in group 2, whereas there were no differences between the control group and group 1. SR(R) and SR(C) correlated well with peak dP/dt (r = 0.71, P <.001 and r = -0.63, P <.001, respectively), whereas SR(L) did not. These findings suggest that LV long axis function becomes depressed earlier than short-axis function in the chronic remodeling process. CONCLUSIONS: Left ventricular short-axis function is a useful marker of LV contractility in patients with chronic, severe MR. Left ventricular long-axis function becomes depressed earlier in the chronic remodeling process. Therefore, evaluation of short-axis as well as long-axis function might be important for better assessment of LV contractile function in these patients. PMID- 19185643 TI - Common endothelial progenitor cell assays identify discrete endothelial progenitor cell populations. AB - BACKGROUND: Multiple measures of endothelial progenitor cells (EPCs) have been described, but there has been limited study of the comparability of these assays. We sought to determine the reproducibility of and correlation between alternative EPC assay methodologies. METHODS: We simultaneously assessed EPC numbers in 140 patients undergoing cardiac catheterization using the 2 most commonly used culture techniques: endothelial cell outgrowth and colony-forming unit (CFU). In the final 77 patients, EPCs were also identified on the basis of cell surface marker expression (CD133, CD34, and vascular endothelial growth factor receptor-2 [VEGFR-2]) and aldehyde dehydrogenase (ALDH) activity. RESULTS: Endothelial progenitor cell enumeration based on fluorescence activated cell sorting was more precise than culture assays. There was limited correlation between EPC numbers determined using the 2 common culture-based assays; however, endothelial CFUs correlated with VEGFR-2 and CD34/VEGFR-2-expressing cells. Endothelial progenitor cells defined by expression of CD133, CD34, CD133/CD34, and ALDH activity correlated with each other, but not with VEGFR-2(+) cells. CONCLUSIONS: Endothelial progenitor cells can be broadly classified into 2 classes: VEGFR-2 expressing cells, which give rise to endothelial CFUs, and CD133/CD34 or ALDH(br) cells. These observations underscore the need for better assay standardization and a more precise definition of EPCs in cell therapy research. PMID- 19185645 TI - Efficacy and safety of the cholesteryl ester transfer protein inhibitor anacetrapib as monotherapy and coadministered with atorvastatin in dyslipidemic patients. AB - BACKGROUND: High-density lipoprotein cholesterol (HDL-C) levels are inversely associated with cardiovascular risk. Cholesteryl ester transfer protein inhibition is one strategy for increasing HDL-C. This study evaluated the lipid altering efficacy and safety of the cholesteryl ester transfer protein inhibitor anacetrapib as monotherapy or coadministered with atorvastatin in patients with dyslipidemia. METHODS: A total of 589 patients with primary hypercholesterolemia or mixed hyperlipidemia (53.8% of the study population had low HDL-C) were randomized equally to one of 10 groups: 5 groups received background statin therapy of atorvastatin 20 mg and 5 did not, and each of these was randomized to placebo, anacetrapib 10, 40, 150, and 300 mg once daily for 8 weeks. An equal proportion of patients had triglycerides >150 mg/dL in each group. RESULTS: For placebo and anacetrapib monotherapy (10, 40, 150, and 300 mg), least squares mean percent changes from baseline to week 8 for low-density lipoprotein cholesterol (LDL-C) were 2%, -16%, -27%, -40%, and -39%, respectively, and for HDL-C were 4%, 44%, 86%, 139%, and 133%, respectively (P < .001 vs placebo for all doses). Coadministration of anacetrapib with atorvastatin produced significant incremental LDL-C reductions and similar HDL-C increases versus atorvastatin monotherapy. For both anacetrapib monotherapy and coadministration with atorvastatin, the LDL-C reductions were similar in patients with baseline triglyceride levels greater than and less than or equal to the median. Anacetrapib was well tolerated, and the incidence of adverse events was similar for placebo and all active treatment groups. There were no increases in systolic or diastolic blood pressure in any treatment arm. CONCLUSIONS: Anacetrapib, as monotherapy or coadministered with atorvastatin, produced significant reductions in LDL-C and increases in HDL-C; the net result of treatment with anacetrapib + atorvastatin was approximately 70% lowering of LDL-C and more than doubling of HDL-C. Anacetrapib was generally well tolerated with no discernable effect on blood pressure. PMID- 19185646 TI - Contrast-enhanced magnetic resonance imaging identifies focal regions of intramyocardial fibrosis in patients with severe aortic valve disease: Correlation with quantitative histopathology. AB - BACKGROUND: Chronic aortic valve disease (AVD) is characterized by progressive accumulation of interstitial myocardial fibrosis (MF). However, assessment of MF accumulation has only been possible through histologic analyses of endomyocardial biopsies. We sought to evaluate contrast-enhanced magnetic resonance imaging (ce MRI) as a noninvasive method to identify the presence of increased MF in patients with severe AVD. METHODS: Seventy patients scheduled to undergo aortic valve replacement surgery were examined by cine and ce-MRI in a 1.5-T scanner. Cine images were used for the assessment of left ventricular (LV) volumes, mass, and function. Delayed-enhancement images were used to characterize the regions of MF. In addition, histologic analyses of myocardial samples obtained during aortic valve replacement surgery were used for direct quantification of interstitial MF. Ten additional subjects who died of noncardiac causes served as controls for the quantitative histologic analyses. RESULTS: Interstitial MF determined by histopathologic analysis was higher in patients with AVD than in controls (2.7% +/- 2.0% vs 0.6% +/- 0.2%, P = .001). When compared with histopathologic results, ce-MRI demonstrated a sensitivity of 74%, a specificity of 81%, and an accuracy of 76% to identify AVD patients with increased interstitial MF. There was a significant inverse correlation between interstitial MF and LV ejection fraction (r = -0.67, P < .0001). Accordingly, patients with identifiable focal regions of MF by ce-MRI exhibited worse LV systolic function than those without MF (45% +/- 14% vs 65% +/- 14%, P < .0001). CONCLUSIONS: Contrast-enhanced MRI allows for the noninvasive detection of focal regions of MF in patients with severe AVD. Moreover, patients with identifiable MF by ce-MRI exhibited worse LV functional parameters. PMID- 19185647 TI - Bleeding risk associated with 1 year of dual antiplatelet therapy after percutaneous coronary intervention: Insights from the Clopidogrel for the Reduction of Events During Observation (CREDO) trial. AB - BACKGROUND: The optimal duration of dual antiplatelet therapy after percutaneous coronary intervention (PCI) is unknown. Incremental reductions in the risk of major adverse cardiovascular events may be partially offset by an increased incidence of bleeding in the months after a PCI. METHODS: We examined the incidence, severity, and predictors of bleeding associated with 1 year of dual antiplatelet therapy after PCI among 1,816 patients in the Clopidogrel for the Reduction of Event During Observation (CREDO) trial. We also compared bleeding in patients who received dual antiplatelet therapy for 1 year to those who did so for only 4 weeks. Bleeding was categorized as major or minor using the modified Thrombolysis In Myocardial Infarction (TIMI) Study Group criteria. RESULTS: Major or minor bleeding occurred in 146 patients during 1 year of follow-up. More than 80% of bleeding events were periprocedural. Multivariable predictors of any bleeding included increasing age and coronary artery bypass. Any (major or minor) bleeding occurred in 71 (8.1%) and 77 (8.9%), major bleeding in 34 (3.9%) and 49 (5.6%), and minor bleeding in 37 (4.2%) and 29 (3.3%) of placebo- and clopidogrel treated patients, respectively; these differences were not significant. However, major gastrointestinal bleeding occurred in significantly more clopidogrel- than placebo-treated patients (13 [1.4%] vs 3 [0.3%] [P = .011]). CONCLUSIONS: Adding clopidogrel to aspirin beyond 4 weeks post PCI is not associated with a significant increase in the overall rate of major or minor bleeding, although it is associated with an increase in major gastrointestinal bleeding in the year after a PCI. PMID- 19185648 TI - Impact of high loading and maintenance dose of clopidogrel within the first 15 days after percutaneous coronary intervention on patient outcome. AB - BACKGROUND: An increase in clopidogrel dose results in an improved inhibition of platelet aggregation. However, whether an increase in clopidogrel dose may improve patient outcome is still debated. The aim of this study was to analyze the impact on patient outcome of an increase in clopidogrel loading and maintenance doses within the first 15 days after percutaneous coronary intervention (PCI). METHODS: Between 2003 and 2007, we included 2,954 consecutive patients who underwent PCI and stent implantation. We compared 2 historical groups. In the "low-dose" group (2003-2005, n = 1,984), patients were pretreated with a 300-mg clopidogrel loading dose followed by 75 mg/d after PCI. In the "high-dose" group (2006-2007, n = 970), patients were pretreated with a 600-mg clopidogrel loading dose followed by 150 mg/d within the first 15 days and 75 mg/d thereafter. The composite primary end point (death, myocardial infarction, stent thrombosis) and bleeding were systematically indexed during the 2-month follow-up period. RESULTS: Clinical and most of angiographic characteristics were similar between the 2 groups. By multivariate analysis, high dose of clopidogrel was associated with a decrease in the composite primary end point (hazard ratio 0.694, 95% CI 0.485-0.993, P = .046). The other predictors were age, left ventricular ejection fraction, diabetes, renal failure, and acute coronary syndrome. Major bleeding was similar in the low- and high-dose groups (2.8% vs 3.4%, respectively, P = .379). After propensity score matching, the high-dose group was still associated with a significant clinical benefit. CONCLUSION: Our results show that a 600-mg loading dose followed by a 150-mg maintenance dose of clopidogrel within the first 15 days after PCI is independently associated with a decrease in the composite death-myocardial infarction-stent thrombosis at 2 months without increase in hemorrhagic complications. PMID- 19185649 TI - Predictive factors for in-stent late loss and coronary lesion progression in patients with type 2 diabetes mellitus randomized to rosiglitazone or placebo. AB - BACKGROUND: Type 2 diabetics (DM2) are at increased risk for restenosis as well as nonculprit coronary artery lesion (NCCL) progression. Rosiglitazone (RSG) favorably modifies many of the altered biologic processes in DM2, although recent reports have questioned its safety. We conducted a double-blind randomized trial to assess the effects of RSG versus placebo on in-stent late lumen loss (LL) and angiographic progression of NCCL. METHODS: A total of 65 DM2 were randomized to RSG (4 mg/d) (n = 32) or placebo (n = 33) at the time of stenting and underwent clinical and laboratory analysis at 1 and 4 months and 8-month angiography (n = 46 patients). Rapid angiographic progression (RAP) was defined as > or =20% diameter reduction of preexisting NCCL by quantitative coronary angiography, or a new narrowing > or =30%. RESULTS: Mean LL in RSG (n = 33 lesions) was not different from that of placebo (0.62 +/- 0.59 vs 0.70 +/- 0.67, P = NS). Seven (13.5%) of 52 NCCLs have RAP in RSG versus 9 (16.1%) of 56 in placebo (P = NS). High-sensitivity C-reactive protein (hs-CRP) was the only predictor of RAP. Patients with a 120-day hs-CRP > or =75th percentile had an OR of 7.35 (95% CI 2.35-23) for RAP versus those below. Although RSG treatment also lowered log (hs CRP) at 4 months (RSG 0.10 +/- 0.37 vs placebo 0.26 +/- 0.49, P = .06), it did not decrease the likelihood of plaque progression while also raising LDL and N terminal brain naturetic peptide. CONCLUSIONS: Rosiglitazone appears not to lower LL or reduce angiographic progression of NCCL in DM2 and had complex effects on markers of cardiac risk. PMID- 19185650 TI - S447X variant of the lipoprotein lipase gene, lipids, and risk of coronary heart disease in 3 prospective cohort studies. AB - BACKGROUND: Lipoprotein lipase (LPL) has a prominent role in the metabolism of triglycerides (TGs) and high-density lipoprotein cholesterol (HDL-C) and is a potential interesting target for the development of antiatherogenic treatment. To provide deeper insight into the role of natural variation in this gene, we investigated the association between the LPL S447X variant with lipids and risk of coronary heart disease (CHD) in 3 independent, prospective studies. METHODS: The S447X variant was genotyped in case-control studies of incident CHD nested within the Nurses' Health Study (NHS), the Health Professionals Follow-up Study (HPFS), and the Danish Diet, Cancer and Health (DCH) study, totaling 245, 258, and 962 cases, respectively. RESULTS: S447X carriers tended to have lower TG and higher HDL-C concentrations than noncarriers. The S447X variant was associated with a lower risk of CHD in the NHS; the association was weaker in the HPFS and not statistically significant in the DCH women and men. The pooled relative risk per minor allele was 0.74 (0.56-1.00). There was a suggestion that the associations of the S447X variant with plasma lipids and CHD risk were more pronounced in obese individuals in the NHS study, but this finding was not consistent across the studies. CONCLUSIONS: The LPL S447X variant tended to be associated with lower TG and higher HDL-C levels, and lower risk of CHD in all 3 cohorts. Lipoprotein lipase is an attractive target for clinical intervention, but studies are needed to clarify whether greater benefit from this variant may be conferred in some subgroups. PMID- 19185651 TI - Competing risk analysis of cause-specific mortality in patients with an implantable cardioverter-defibrillator: The EVADEF cohort study. AB - BACKGROUND: Although implantable cardioverter-defibrillator (ICD) therapy has been evaluated in randomized controlled trials, enrolling highly selected patients, mortality events in ICD patients have received little attention in routine medical care. We sought to assess the 24-month total and cause-specific mortality rates and their predictors in "real life" patients with an ICD. METHODS: The Evaluation Medico-Economique du Defibrillateur Automatique Implantable study was a French multicenter, prospective, observational cohort study of ICD patients with a 2-year follow-up. Cause-specific mortality rates and predictors at implantation of sudden cardiac death (SCD) or progressive heart failure (HF) death were assessed using competing risk methodology. RESULTS: From June 2001 to June 2003, 2,296 unselected patients were implanted and followed until June 2005. During a mean follow-up of 20.5 +/- 6.7 months, 274 deaths occurred: 29 (10.6%) were SCD and 146 (53.3%) were HF deaths, corresponding to 24 month cause-specific mortality rates of 1.4% (95% confidence interval 0.9%-1.9%) and 6.9% (95% confidence interval 5.8%-8.0%), respectively. Among the characteristics at implantation, ejection fraction (EF) <30% and history of atrial fibrillation were independently associated with SCD; age, high New York Heart Association class, systemic hypertension, prior atrial fibrillation, QRS duration, EF <30%, and lack of beta-blocker therapy were independently associated with HF death. CONCLUSIONS: In this large cohort of "daily" patients, the 2-year incidence of SCD (1.4%) was comparable with the event rate observed in randomized controlled trials; HF remained the predominant mode of death. An EF <30% at implantation appears to be the most important predictor of ICD-unresponsive SCD. PMID- 19185652 TI - Established, emerging and future roles of PET/CT in the management of colorectal cancer. AB - Positron-emission tomography-computed tomography (PET/CT) is rapidly being integrated into the imaging pathways of several different tumour types, most frequently using the glucose analogue 2- [(18)F]-fluoro-2-deoxy-D-glucose (FDG). Integrated FDG-PET/CT combines functional and anatomical imaging to improve sensitivity and specificity of tumour detection. The aim of this article is to review the established, emerging, and future roles of FDG-PET/CT in the management of patients with colorectal cancer (CRC). PMID- 19185653 TI - Chronic left ventricular failure: the role of imaging in diagnosis and planning of conventional and novel therapies. AB - Heart failure is the leading cause of hospitalisation in the UK, and its prevalence is expected to increase further in the future due partly to an aging population. Although pharmacological agents remain the mainstay of therapy, an increasing number of surgical and novel minimally invasive interventions have been developed for the treatment of both acute and chronic heart failure. Imaging is essential for diagnosis, guiding therapeutic options, and monitoring therapy and its complications. As a result, radiologists should be familiar with the pathogenesis, treatment options, and imaging-related issues pertaining to the management of these patients. PMID- 19185654 TI - Incremental detection of coronary artery disease by assessment of non-calcified plaque on coronary CT angiography. AB - AIM: The purpose of this study was to evaluate the improved assessment of coronary atherosclerotic plaque burden by measurement of non-calcified plaque in addition to calcified plaque using CT coronary angiography (CTA). MATERIALS AND METHODS: Low to intermediate-risk outpatients with suspected coronary artery disease were prospectively recruited. Patients underwent CTA and calcium scoring in addition to invasive angiography. The presence of plaque (calcified, non calcified, and mixed) was analysed on a per segment basis (percentage of segments with disease) with stratification by calcium score (CS). RESULTS: Seventy-six patients were enrolled of whom 30 had a CS of 0, 26 had a CS of 1-200, and 20 had a CS of >200. One thousand, one hundred and two segments were analysed using CTA and invasive angiography. The prevalence of segments with calcified or mixed plaque was 3.1% (n=13) for a CS of 0, 15.1% (n=57) for a CS of 1-200, and 50% (n=142) for a CS of >200 (all p<0.0001). The proportion of segments with non calcified plaque alone was low and similar among the three groups: 5.4% (n=23; CS=0), 8.2% (n=32; CS=1-200), and 8.6% (n=25; CS= >200), (CS=0 versus CS= >200; p=0.04, others p=ns). The relative increase in diseased segments by additional assessment of non-calcified plaque was greatest for patients with a CS of 0 (173%) versus a CS of 1-200 (55%), and a CS of >200 (17%). CONCLUSION: CTA offers increased relative incremental detection of non-calcified plaque, particularly in those with negative CS; however, the absolute detection of non-calcified plaque in those with negative CS is low. The prognostic significance of non-calcified plaque for the prediction of cardiac events, particularly in patients with low CS, requires continued study. PMID- 19185655 TI - Atypical teratoid/rhabdoid tumors of the central nervous system: imaging and clinical findings in 16 children. AB - AIM: To investigate the imaging and clinical findings of central nervous system (CNS) atypical teratoid/rhabdoid tumours (AT/RTs) in children. MATERIALS AND METHODS: The computed tomography (CT) and magnetic resonance imaging (MRI) findings and clinical records of 16 children with CNS AT/RTs were retrospectively reviewed. Tumour location, size, composition, enhancement pattern, peritumoural oedema, signal intensity (SI) on MRI and CT attenuation were evaluated. RESULTS: A total of 17 lesions from 16 patients (median age 2.3 years, age range 0.7-15 years) were included in the evaluation. Tumour location was infratentorial for 11 lesions and supratentorial for six lesions. The mean diameter of the largest dimension for a tumour was 4 cm. The tumour was mainly solid in 65% of cases, and solid and cystic or cystic and solid in 35% of cases. The solid component of the tumours had a homogeneous iso SI (n=15) on T2-weighted MRI images and iso SI (n=14) on T1-weighted images. Moderate to strong enhancement of the solid component was noted in most cases. In spite of a large tumour size, peritumoural oedema was minimal or mild except in four cases. Rapid growth of the tumour was demonstrated in three cases. Seven patients died from tumour progression, with a mean survival time of 8.4 months (range 2-12 months). CONCLUSION: Although the AT/RTs had non-specific imaging findings, the tumours tended to be large in size, have iso SI on T1 and T2-weighted MR images with prominent enhancement, and relatively mild peritumoural oedema. Rapid growth of the tumour was seen during the follow-up period. PMID- 19185656 TI - Oesophageal strictures caused by the ingestion of corrosive agents: effectiveness of balloon dilatation in children. AB - AIM: To evaluate the safety and clinical effectiveness of balloon dilatation in children for oesophageal strictures caused by the ingestion of corrosive agents. MATERIALS AND METHODS: The study comprised 11 children (median age 6 years; range 1-14 years) with oesophageal strictures caused by corrosive agents, who underwent a total of 36 balloon dilatation sessions. The technical and clinical success, recurrence of dysphagia, complications, and primary and secondary patency rates were retrospectively evaluated. RESULTS: Technical success was achieved in 91% of patients and in 97% of balloon dilatation sessions. Clinical success (defined as improved food intake and reduced dysphagia within 1 month of the first balloon dilatation session) was achieved in 64% of patients (7/11). During the mean 35 month follow-up period (range 1-89 months), 10 (91%) patients experienced recurrence. Oesophageal rupture (types 1 or 2) occurred in 45% of patients and in 31% of balloon dilatation sessions. Primary patency rates at 6 months and 1, 2, 3, 4, and 5 years were 36, 27, 14, 14, 14, and 14%, respectively. Secondary patency rates at 6 months and 1, 2, 3, 4, and 5 years were 82, 82, 82, 56, 42, and 42%, respectively. The secondary patency rate was higher than the primary patency rate (p<0.05). CONCLUSION: The present study examined oesophageal balloon dilatation for paediatric oesophageal strictures caused by the ingestion of corrosive agents. Although the technical success rate was high and there were no deaths, the clinical success rate was low owing to a high recurrence rate. However, repeated balloon dilatations resulted in an acceptable secondary patency rate. PMID- 19185657 TI - Predictors of abnormal chest CT after blunt trauma: a critical appraisal of the literature. AB - AIM: To identify and to evaluate predictors that determine whether chest computed tomography (CT) is likely to reveal relevant injuries in adult blunt trauma patients. METHODS: After a comprehensive literature search for original studies on blunt chest injury diagnosis, two independent observers included studies on the accuracy of parameters derived from history, physical examination, or diagnostic imaging that might predict injuries at (multidetector row) CT in adults and that allowed construction of 2x2 contingency tables. For each article, methodological quality was scored and relevant predictors for injuries at CT were extracted. For each predictor, sensitivity, specificity, positive and negative likelihood ratio and diagnostic odds ratio (DOR) including 95% confidence intervals were calculated. RESULTS: Of 147 articles initially identified, the observers included 10 original studies in consensus. Abnormalities at physical examination (abnormal respiratory effort, need for assisted ventilation, reduced airentry, coma, chest wall tenderness) and pelvic fractures were significant predictors (DOR: 2.1-6.7). The presence of any injuries at conventional radiography of the chest (eight articles) was a more powerful significant predictor (DOR: 2.2-37). Abnormal chest ultrasonography (four articles) was the most accurate predictor for chest injury at CT (DOR: 491-infinite). CONCLUSION: The current literature indicates that in blunt trauma patients with abnormal physical examination, abnormal conventional radiography, or abnormal ultrasonography of the chest, CT was likely to reveal relevant chest injuries. However, there was no strong evidence to suggest that CT could be omitted in patients without these criteria, or whether these findings are beneficial for patients. PMID- 19185658 TI - National audit of provision of MRI services 2006/07. AB - In 2003 the Royal College of Radiologists Clinical Radiology Audit Sub-Committee began an audit process evaluating the standards of provision of magnetic resonance imaging (MRI) services. This was prompted by the publication of the 2002 Audit Commission Report, which had identified that lack of MRI provision was responsible for more than half of the total waiting times for diagnostic imaging investigations. The audit found that the time from request to report did not meet the standard for cancer staging examinations, but nationally, was within the target set for routine orthopaedic examinations. However, national mean waiting times were longer than recommended for both cancer and orthopaedic MRI. Since then, there has been massive investment in MRI capacity, both from installation of MRI systems in NHS Trusts, and in England, from outsourcing of routine MRI cases through the Department of Health contract with an independent provider. A re-audit in 2006/7 shows that there has been a significant improvement in waiting times for routine orthopaedic examinations, but the position with cancer staging examinations has deteriorated. Control chart methodology shows that underperformance is due to common cause variation, i.e., improvements need to be made to the overall process from receiving the request for MRI to the issue of the report. Follow-up with participating departments demonstrated there were some common themes for underperformance, and suggestions for improvement are made from departments with best performance. PMID- 19185659 TI - Which oblique plane is more helpful in diagnosing an anterior cruciate ligament tear? AB - AIM: To evaluate the diagnostic role of additional oblique coronal and oblique sagittal magnetic resonance imaging (MRI) for an anterior cruciate ligament (ACL) tear. MATERIALS AND METHODS: A total of 101 patients who had undergone preoperative knee MRI examinations with orthogonal and two sets of oblique images were enrolled in the study. Two radiologists evaluated the MRI images by the use of four methods: orthogonal images only (method A); orthogonal and additional oblique coronal images (method B); orthogonal and oblique sagittal images (method C); and orthogonal images with oblique coronal and sagittal images (method D). The status of the ACL (normal or tear) was determined by consensus. The sensitivity, specificity, and accuracy for an ACL tear with the use of each method were calculated in comparison with arthroscopy as the reference standard, and values were statistically analysed using the McNemar test. The diagnostic accuracies were compared using receiver operating characteristic (ROC) analysis. RESULTS: Arthroscopy identified 10 partial ACL tears and 30 complete ACL tears. The specificities and accuracies for methods B, C, and D were significantly higher than the specificities and accuracies for method A (p<0.05). There was no significant difference in the sensitivity, specificity, and accuracy for methods B, C, and D. Diagnostic ability was not significantly different for each method, as determined by ROC analysis (p>0.05). CONCLUSIONS: Additional oblique imaging for an ACL tear improved the specificity. Either of the oblique imaging methods is sufficient, and no further improvement in the diagnostic efficacy was achieved by simultaneous use. PMID- 19185660 TI - Symmetry in computed tomography of the brain: the pitfalls. AB - Computed tomography (CT) studies of the brain are one of the most frequent examinations interpreted by radiologists out of hours. Apparently normal appearances in patients with significant neurological morbidity can be perplexing. As the contents of the cranium are normally remarkably symmetrical on axial CT, disease entities that result in symmetrical appearances are the most difficult to detect. In this review we highlight a spectrum of important acute neurological conditions that result in abnormal but symmetrical appearances on CT. PMID- 19185661 TI - MRI of impingement syndromes of the shoulder. AB - The diagnosis of shoulder impingement is primarily a clinical one. Imaging has a role in assisting clinicians in developing a treatment strategy by identifying and characterizing the cause of shoulder impingement. In this review, the relevant anatomy, cause/pathomechanics, clinical features, and magnetic resonance imaging (MRI) findings of the different types of impingement syndromes are presented. PMID- 19185662 TI - Imaging in pulsatile tinnitus. AB - Tinnitus may be continuous or pulsatile. Vascular lesions are the most frequent radiologically demonstrable cause of pulsatile tinnitus. These include congenital vascular anomalies (which may be arterial or venous), vascular tumours, and a variety of acquired vasculopathies. The choice of imaging depends on the clinical findings. If a mass is present at otoscopy, thin-section computed tomography (CT) is indicated. In the otoscopically normal patient, there is a range of possible imaging approaches. However, combined CT angiography and venography is particularly useful. PMID- 19185663 TI - Diagnosis of hepatic angiomyolipomata using CT: report of three cases and review of the literature. PMID- 19185664 TI - MR cholangiopancreatographic demonstration of biliary tract abnormalities in AIDS cholangiopathy: report of two cases. PMID- 19185665 TI - Re: Investigating suspected acute pulmonary embolism--what are hospital clinicians thinking? PMID- 19185666 TI - A 21st-century Trojan horse: the "abortion harms women" anti-choice argument disguises a harmful movement. PMID- 19185667 TI - Depo-Provera and skeletal health: reviewing the evidence; developing and disseminating a consensus. PMID- 19185668 TI - The use of depot-medroxyprogesterone acetate in contraception and its potential impact on skeletal health. AB - BACKGROUND: In the fall of 2007, the controversy about the contraceptive use of depot-medroxyprogesterone acetate (DMPA) and its potential impact on skeletal health reached the media in the province of Quebec, Canada, thereby becoming a matter of concern for the lay public and physicians. In order to discuss this subject openly, the National Institute of Public Health of Quebec (INSPQ) organized a scientific meeting on February 15, 2008, with targeted physicians delegated by their medical associations in the fields of general practice, obstetrics and gynaecology, rheumatology, orthopaedic surgery, physiatry and endocrinology. STUDY DESIGN: Participants reviewed the scientific literature using the study classification method according to the level of evidence, reviewed published guidelines of medical societies and organizations on the subject and reached a consensus position. This manuscript presents a review of the literature and describes the consensus position of the targeted medical associations. RESULTS: The consensus position adopted by all the targeted medical associations determined that DMPA was a cost-effective contraceptive option that must be considered in the light of the clinical situation and preference of each woman. Candidates for injectable contraception should be informed that the use of DMPA is associated with a slight decrease in bone mineral density (BMD), which is largely, if not completely, reversible. There should not be an absolute limit to the length of time that the DMPA contraceptive is used, regardless of the woman's age. Monitoring BMD is not recommended among users of DMPA for contraceptive purposes. Finally, the consensus statement declared that, although supplements of calcium and vitamin D are beneficial for skeletal health for women in general, such supplementation should not be recommended solely based on a woman's use of DMPA. CONCLUSION: Given the scientific evidences, DMPA use remains a valid contraceptive option for women. Its potential impact on BMD must be balanced against the significant individual, familial and social consequences of unintended pregnancy. PMID- 19185669 TI - Contraceptive failure related to estimated cycle day of conception relative to the start of the last bleeding episode. AB - BACKGROUND: The objective of this study was to estimate the menstrual cycle day of conception in women presenting for abortion. STUDY DESIGN: This was a retrospective chart survey in two urban free-standing abortion clinics. RESULTS: There were 913 charts reviewed of women presenting for an abortion at less than 63 days' gestation as determined by endovaginal ultrasound who were "sure" of the date of their last normal menstrual period. The estimated mean cycle day of conception determined by sonographically estimating length of gestation was 14.6. There were 26 (26.3%) of 99 women using cyclic hormonal contraception who conceived before 10 days after the onset of withdrawal bleeding compared to 100 (14.7%) of 679 who conceived before 10 days after the onset of their last menstrual period who were using all other forms of contraception, including "none" (p=.005). No other differences in the proportions conceiving early in the cycle were observed with respect to age, ethnicity or obesity. CONCLUSION: These data suggest that there is a sizeable subset of women who ovulate earlier after onset of withdrawal bleeding when using 21/7 hormonal contraceptives than after onset of menses when not using hormonal contraception. It is possible that women using hormonal contraceptives may have a higher risk of pregnancy if they ovulate sooner after the onset of bleeding. PMID- 19185670 TI - Follicular development in a 7-day versus 4-day hormone-free interval with an oral contraceptive containing 20 mcg ethinyl estradiol and 1 mg norethindrone acetate. AB - BACKGROUND: Combined oral contraceptive (COC) formulations with 20 mcg ethinyl estradiol (EE) have a greater incidence of ovarian hormone production and follicular development, which can be managed by shortening the number of hormone free days per COC cycle. This study evaluates differences in follicular development during a 7-day versus 4-day hormone-free interval in a COC regimen with 20 mcg EE and 1 mg norethindrone acetate. STUDY DESIGN: Forty-one healthy women were randomized in an open-label fashion to this formulation in either a 24/4 or a 21/7 day regimen for three cycles. Estradiol, progesterone, follicle stimulating hormone, luteinizing hormone and inhibin B were measured daily from Cycle 2, Day 21 to Cycle 3, Day 3 and on Day 7 of Cycle 3. Follicular diameter and Hoogland score were calculated on Cycle 2, Days 21, 24 and 28 and Cycle 3, Days 3 and 7. RESULTS: Sixty-six percent of subjects in the 21/7 group and 70% of the subjects in the 24/4 group developed a follicle greater than 10 mm diameter. Ovarian steroid hormone levels, Hoogland scores and bleeding patterns were not statistically significant between the groups. CONCLUSION: In contrast to prior studies, this analysis suggests no difference in follicle development or bleeding patterns among women receiving a 21/7 or 24/4 regimen of a 20-mcg EE/1-mg norethindrone acetate COC. PMID- 19185671 TI - The LNG-IUS study on adenomyosis: a 3-year follow-up study on the efficacy and side effects of the use of levonorgestrel intrauterine system for the treatment of dysmenorrhea associated with adenomyosis. AB - BACKGROUND: The objective of this study was to evaluate the efficacy and side effects of the levonorgestrel-releasing intrauterine system (LNG-IUS) in the treatment of moderate or severe dysmenorrhea associated with adenomyosis for a 3 year follow-up period. STUDY DESIGN: The LNG-IUS was inserted into 94 women who had moderate or severe dysmenorrhea associated with adenomyosis diagnosed by transvaginal sonography during Cycle Days 5-7. A visual analogue scale (VAS) of dysmenorrhea, uterine volume and serum CA125 levels were used to assess the efficacy of the treatment at baseline and 3, 6, 12, 24 and 36 months after the LNG-IUS insertion. Side effects were recorded at every follow-up visit. RESULTS: The VAS of dysmenorrhea dropped continuously and significantly from the baseline score of 77.9+/-14.7 to 11.8+/-17.9 after 36 months of the LNG-IUS insertion (p<.001). The uterine volume decreased significantly from 113.8+/-46.9 mL to 94.5+/-40.1 mL (p=.003) at 6 months and to 87.7+/-35.8 mL (p<.001) at 12 months and then rose slightly, but the variables at 24 and 36 months still decreased significantly in comparison with the baseline variable (p<.001). The serum CA125 levels reduced significantly starting from 6 months after device insertion (p<.001). The most common side effects were weight gain (28.7%), simple ovarian cyst formation (22.3%) and lower abdominal pain (12.8%). At 36 months, the overall satisfaction rate of the treatment was 72.5%. CONCLUSIONS: The LNG-IUS appears to be an effective method in alleviating dysmenorrhea associated with adenomyosis during 3 years. It may be a valuable long-term alternative for the treatment of adenomyosis. PMID- 19185672 TI - Unintended pregnancy and associated maternal preconception, prenatal and postpartum behaviors. AB - BACKGROUND: This study was conducted to determine the relationship between unintended pregnancy and maternal behaviors before, during and after pregnancy. STUDY DESIGN: Data were analyzed from a stratified random sample of 9048 mothers who delivered live born infants between 2001 and 2006 and completed the Pregnancy Risk Assessment Monitoring System (PRAMS) survey 2 to 9 months after delivery. Binary and ordinal logistic regression methods with appropriate survey weights were used to control for socio-demographic factors. RESULTS: Compared to women with intended pregnancies, mothers with unwanted pregnancies were more likely to consume less than the recommended amount of preconception folic acid [adjusted odds ratio (OR) 2.39, 95% confidence interval (CI) 1.7-3.2], smoke prenatally (OR 2.03, 95% CI 1.5-2.9), smoke postpartum (OR 1.86, 95% CI 1.35-2.55) and report postpartum depression (OR 1.98, 95% CI 1.48-2.64); they were less likely to initiate prenatal care during the first trimester (OR 0.34, 95% CI 0.3-0.5) and breastfeed for 8 or more weeks (OR 0.74, 95% CI 0.57-0.97). Compared to women with intended pregnancies, women with mistimed pregnancies were also more likely to consume inadequate folic acid, delay prenatal care and report postpartum depression. CONCLUSION: Even after controlling for multiple socio-demographic factors, unwanted and mistimed pregnancies were associated with unhealthy perinatal behaviors. PMID- 19185673 TI - Comprehension of a prototype emergency contraception package label by female adolescents. AB - BACKGROUND: We evaluated female adolescents' comprehension of a prototype over the-counter package label for an emergency contraceptive pill product. STUDY DESIGN: Volunteers aged 12-17 years who could read English were recruited at malls and clinics in six United States metropolitan areas. After completing a literacy assessment, subjects examined the prototype package and answered 20 questions that assessed understanding of six key concepts related to appropriate use of the product. RESULTS: The analysis population included 335 subjects, 54 to 59 of each year of age between 12 and 17 years. When asked what the product is used for, 264 respondents (79%) specifically indicated contraception. The six key concepts were each understood by 83-96% of subjects. In all 24 population subgroups examined, each key concept was understood by at least 72% of subjects. CONCLUSION: Female adolescents aged 17 years and younger understand the prototype package label well enough to enable safe and effective use without assistance from a clinician. PMID- 19185674 TI - Accuracy of information given by Los Angeles County pharmacies about emergency contraceptives to sham patient in need. AB - BACKGROUND: As emergency contraception (EC) becomes increasingly available without prescription, it is important that women in need be provided accurate information about its use. STUDY DESIGN: A telephone survey of all retail pharmacies in Los Angeles County was conducted by women posing as a 23-year-old who wanted to prevent pregnancy after an unprotected act of intercourse. RESULTS: One thousand four hundred sixty unduplicated pharmacies listed in the Yellow Pages 2007 were called between October 2007 and April 2008. Sixty-nine percent had EC available on site; 19% referred the caller elsewhere. The remainder said nothing could be done or hung up. Multiple calls and multiple recitations of the situation were needed in over one third of calls. Isolated incidents of inappropriate comments occurred. CONCLUSION: While most pharmacies provided information about EC that was consistent with labeling, barriers still exist to both accurate information and timely access to that product. PMID- 19185675 TI - Do instructions for over-the-counter pre-coital female contraceptives promote "perfect use"? AB - BACKGROUND: This study was conducted to estimate the readability and related features of English-language over-the-counter (OTC) pre-coital female contraceptive (PFC) instructions. STUDY DESIGN: We identified and purchased all currently available OTC PFCs (n=8), including Encare (Contraceptive Gel and Insert), F.C. Female Condom, Ortho Options (Conceptrol, Delfen and Gyncol II Jelly), Today Sponge and VCF Vaginal Film. Reading grade level was calculated using the Simple Measure of Gobbledygook. Text point size was measured and total number of graphics was tallied. Graphic dimensions were also measured. We also assessed OTC PFC instructions on four main criteria--derived from the "User Friendliness Tool (UFT)"--including layout features, graphic characteristics, presence of a clear message and presentation of manageable information. RESULTS: Reading level ranged from 8th to 12th grade (mean+/-SD=10.0+/-1.2). Mean page length was 24.4+/-7.0 cm (9.6+/-2.76 in.), while average page width was 20.3+/ 13.8 cm (7.99+/-5.43 in.). Average text point size was 7.9+/-2.3 (range=6-12). Illustrations, predominantly line drawings, were used throughout (range=3-11), and most were similar in size to a matchbox. None of the "How to Use" sections scored high in all criteria on the UFT. CONCLUSIONS: "How to Use" sections of OTC PFC instructions should be revised to be easier to read and more user friendly. Ideally, the gap between "typical" and "perfect" contraceptive efficacy could potentially be narrowed if instructions were developed that the large majority of women could easily understand. PMID- 19185676 TI - Barriers associated with the failure to return for intrauterine device insertion following first-trimester abortion. AB - BACKGROUND: The aim of the study was to identify barriers associated with the failure to return for delayed intrauterine device (IUD) insertion postabortion. STUDY DESIGN: This study had two components: (a) a retrospective cohort study of women who chose an IUD as their postabortion contraceptive method to compare characteristics of those who did and did not receive a device within 6 months of the procedure and (b) a prospective survey of women intending IUD use postabortion to assess actual IUD use at 6 weeks, barriers to access and attitudes on insertion timing. RESULTS: Of the 500 abortion patient charts available for review during the study period, 53 women intended IUD use postabortion. At 6 months, only 32% intending an IUD received one, and there were no significant demographic differences between the groups. For the prospective portion, the response rate at 6 weeks was 54% (27/50), with only 26% (7/27) of responders reporting IUD insertion by this time. The principal reported barrier to IUD insertion was time needed for an additional visit (41%). Most women (67%) desired immediate insertion and believed that they would be more likely to have an IUD inserted if it is performed immediately postabortion (63%). Monthly income was the only statistically significant difference between those who responded to the 6-week follow-up (US$1409.50) and those who did not (US$937.50, p=.05). CONCLUSION: A significant number of women that express a preference for IUD use after first-trimester abortion do not return to obtain a device. Most would prefer to have the option of immediate insertion. PMID- 19185678 TI - Young unmarried men's understanding of female hormonal contraception. AB - BACKGROUND: A better understanding of men's attitudes, norms and behaviors regarding women's use of hormonal contraception is needed. STUDY DESIGN: We conducted contraceptive life-history interviews with 41 ethnically diverse males ages 18-25 years which detailed up to six heterosexual relationships, focusing on knowledge, attitudes, norms and behaviors regarding hormonal contraception use, decision making and communication. RESULTS: Men's attitudes, norms and behaviors associated with hormonal contraceptive decisions and use varied greatly across participants and their relationships. Findings suggest a mixture of attitudes and practices regarding the importance of communication around contraception influenced by sexual experiences, age and relationship type. Many men demonstrated limited knowledge about contraceptives and identified improving contraceptive knowledge as an essential step in facilitating contraceptive communication. CONCLUSIONS: Increased awareness about young men's understanding of and perceived roles regarding hormonal contraception will help in designing services that address contraceptive adherence, contraceptive communication and incorrect or inadequate contraceptive knowledge. PMID- 19185679 TI - Effect of etonogestrel contraceptive implant (Implanon) on portal blood flow and liver functions. AB - BACKGROUND: This study was conducted to evaluate changes in portal blood flow and liver functions among women using Implanon for 2 years. STUDY DESIGN: Fifty healthy Implanon users were enrolled in this longitudinal study and followed up for 24 months. Portal blood flow, assessed by color Doppler; prothrombin time and concentration; serum albumin; bilirubin; gamma-glutamyl transferase (GGT); alanine aminotransferase (ALT); and aspartate aminotransferase (AST) were measured before and 24 months after insertion. RESULTS: After 24 months of Implanon insertion, there were no significant changes in portal blood flow, serum albumin, prothrombin time or concentration. However, there was a significant increase in serum levels of total and unconjugated bilirubin and GGT and a significant decrease in ALT and AST levels. All levels, however, remained within the normal range of values. CONCLUSIONS: Implanon use for 2 years does not seem to influence portal hemodynamics. Changes in serum levels of bilirubin, GGT, ALT and AST are unlikely to be of clinical significance. PMID- 19185677 TI - Unmet need for contraception among sex workers in Madagascar. AB - BACKGROUND: The study was conducted to investigate past and future pregnancy preferences and contraceptive need among Malagasy sex workers. STUDY DESIGN: We analyzed data on pregnancy and contraceptive use collected during the baseline visit of a randomized, prospective formative trial which assessed diaphragm and microbicide acceptability among sex workers. To be eligible, women could not be pregnant or planning pregnancy for the next 2 months. RESULTS: Women (N=192) from four cities (Antananarivo, Antsiranana, Mahajanga and Toamasina) reported a median of 10 sex acts per week. Fifty-two percent reported a prior unwanted pregnancy, 45% at least one induced abortion and 86% that preventing future pregnancy was moderately to very important. During the last sex act, 24% used a hormonal method, 36% used a male condom, 2% used a traditional method and 38% used no method. Nearly 30% of participants reported that pregnancy prevention was moderately or very important but used no contraception at last sex; these women were categorized as having "unmet need" for contraception. In multivariable binomial regression analyses, factors associated with unmet need included low knowledge of contraceptive effectiveness [age- and site-adjusted prevalence ratio (PR): 2.1; 95% confidence interval (CI): 1.4-3.0] and low self-efficacy to negotiate condom use (age- and site-adjusted PR: 2.0; 95% CI: 1.4-3.0). CONCLUSIONS: Among these women, prior unwanted pregnancy and induced abortion were common and preventing future pregnancy was important, yet gaps in contraceptive use were substantial. Contraceptive knowledge and self-efficacy should be improved to promote contraceptive use by sex workers. PMID- 19185680 TI - California Teachers Study report on incomplete pregnancy and breast cancer risk is flawed. PMID- 19185682 TI - Re: Estrogen-progestin contraceptive use during adolescence prevents bone mass acquisition: a 4-year follow-up study. PMID- 19185684 TI - Quality in EUS: an assessment of baseline compliance and performance improvement by using the American Society for Gastrointestinal Endoscopy-American College of Gastroenterology quality indicators. AB - BACKGROUND: An American Society for Gastrointestinal Endoscopy-American College of Gastroenterology (ASGE-ACG) task force recently developed quality indicators for the preprocedure, intraprocedure, and postprocedure phases of each endoscopic procedure. Benchmark rates and clinical significance of compliance have not been determined. OBJECTIVES: To establish baseline compliance rates to the preprocedure and intraprocedure quality indicators in our EUS cases, identify indicators with the lowest compliance rates, and establish change in compliance rates with a targeted performance improvement plan. METHODS: We measured baseline compliance to each of the preprocedure and intraprocedure EUS quality indicators in the EUS procedures performed at Mayo Clinic Jacksonville from March 1996 through August 2006. We developed a performance improvement plan that targeted the 4 indicators with the lowest compliance over the entire time period. Compliance rates in the year after plan implementation were compared with those from January 2004 to August 2006, when adjusting for endoscopist and direct access. RESULTS: We demonstrated areas of high quality as well as areas for improvement in compliance with the ASGE-ACG quality metrics in a large cohort of EUS cases. We achieved improvement in all 4 areas targeted for quality improvement, statistically significant at the 5% level for two of the quality indicators. LIMITATIONS: Limitations included our retrospective design and the use of unstructured procedure dictations that may limit application of our results. It is also unclear whether compliance was truly synonymous with performance. CONCLUSIONS: We established reference levels of compliance rate within our practice and showed that a targeted performance improvement plan that consisted of awareness, individual accountability, and documentation can result in improvement. PMID- 19185685 TI - Improving the quality of EUS: what is the template for success? PMID- 19185686 TI - On the way toward quantitative staining techniques? PMID- 19185687 TI - Prospective randomized trial of EUS versus ERCP-guided common bile duct stone removal: an interim report (with video). AB - BACKGROUND: EUS is being increasingly utilized for the diagnosis of choledocholithiasis and microlithiasis, especially in patients with biliary colic. Simultaneously, there is also a rising interest in the use of EUS for therapeutic interventions. OBJECTIVES: Our goal was to assess the effectiveness of EUS-directed common bile duct (CBD) stone removal to compare its safety and effectiveness with ERCP-directed intervention. DESIGN: Interim results of a prospective, randomized, single-center blinded clinical trial. SETTING: A single tertiary care referral center. PATIENTS: Fifty-two patients with uncomplicated CBD stones were prospectively randomized to CBD cannulation and stone removal under EUS or ERCP guidance. MAIN OUTCOME MEASUREMENTS AND INTERVENTIONS: Primary outcome measure was the rate of successful cannulation of the CBD. Secondary outcome measures included successful removal of stones and overall complication rates. RESULTS: CBD cannulation followed by stone extraction was successful in 23 of 26 patients (88.5%) in the EUS group (I) versus 25 of 26 patients (96.2%) in the ERCP group (II) (95% CI, -27.65%, 9.88%). Overall, there were 3 complications in the EUS group and 4 complications in the ERCP group. LIMITATION: The current study is an interim report from a single center report and performed by a single operator. CONCLUSIONS: Our preliminary analysis indicates that outcomes following EUS-guided CBD stone retrieval are equivalent to those following ERCP. EUS related adverse events are similar to those following ERCP. ERCP and EUS-guided stone retrieval appears to be equally effective for therapeutic interventions of the bile duct. Additional studies are required to validate these preliminary results and to determine predictors of success of EUS-guided stone removal. PMID- 19185688 TI - A novel capsule endoscope: do we need new kids on the block? PMID- 19185689 TI - The on-call fellow--answering the after-hours call. PMID- 19185690 TI - Systematic review of narrow-band imaging for the detection and differentiation of abnormalities in the esophagus and stomach (with video). PMID- 19185691 TI - A new injectable agent for the treatment of GERD: results of the Durasphere pilot trial. AB - BACKGROUND: A variety of endoscopic modalities have been introduced to treat GERD, including radiofrequency energy, suturing, plication, and injection therapy. OBJECTIVE: Our purpose was to assess the long-term safety and effectiveness of Durasphere (Carbon Medical Technologies, St Paul, Minn), a new injectable bulking agent, in the treatment of mild-moderate GERD. DESIGN: Initial human pilot study. SETTING: Single endoscopy center; study period from January 2005 to August 2006. PATIENTS: Ten GERD subjects, confirmed by pH monitoring, on daily proton pump inhibitor (PPI) therapy, hiatal hernia <3 cm, and no or mild erosive esophagitis. INTERVENTION: Endoscopic injection with Durasphere, a new submucosal bulking agent, at the gastroesophageal junction. OUTCOMES: Change in symptom scores, PPI use, pH scores, and endoscopic findings; monitoring of safety profile. RESULTS: Nine of 10 patients completed the 12-month trial. There were no adverse events. The procedure was well tolerated with minimal patient discomfort and no dysphagia. At 12 months 70% of patients discontinued all antacid medication completely; 90% of patients reduced PPI use by greater than 50%. DeMeester scores improved from a mean of 44.5 at baseline to 26.5 at 12 months; 4 patients achieved normal pH scores. There was no esophagitis at 12 months, and no erosion, ulceration or sloughing of material was noted at any injection site. The Durasphere material did not appear to migrate. LIMITATIONS: Nonrandomized study design without a control group; small number of subjects. CONCLUSIONS: Durasphere appears to be a promising new injectable bulking agent for the treatment of mild moderate GERD, with demonstrable efficacy and no significant adverse events in a small cohort. PMID- 19185692 TI - Injectable treatment for GERD: the flight of the Phoenix? PMID- 19185693 TI - Spiral enteroscopy: a new twist on overtube-assisted endoscopy. PMID- 19185694 TI - Pancreatic Schwannoma presenting as a cystic lesion. PMID- 19185695 TI - Rectal perforation in ulcerative colitis: complication of an enema tip. PMID- 19185696 TI - A European case series of endoscopic submucosal dissection for gastric superficial lesions. AB - BACKGROUND: EMR is an accepted method for resection of superficial lesions in the GI tract. However, because it leads, not unusually, to piecemeal resection, histopathologic interpretation problems and an increased risk of recurrence are noticeable. Endoscopic submucosal dissection (ESD) allows a higher rate of en bloc resection, with low recurrence. Nevertheless, this technique, namely in the upper-GI tract, has rarely been described in Western countries, probably because of the rarity of gastric cancer in most countries. OBJECTIVE: To describe the efficacy and safety of ESD for gastric superficial lesions in a European country. DESIGN: Consecutive case-series report. SETTING: A tertiary specialized center. PATIENTS: Nineteen patients with gastric superficial lesions (15-30 mm), with high-grade (n = 15) or low-grade (n = 4) noninvasive epithelial neoplasias, in the antrum (n = 12), incisura angularis (n = 2), body (n = 3), and cardia (n = 2). INTERVENTION: ESD with the patient under general anesthesia in the endoscopic room (40-300 minutes) by using an insulated-tip-knife. MAIN OUTCOME MEASUREMENTS: Complete (R0) and en bloc resection, and complications. RESULTS: ESD was achieved in all cases, with 89% R0 resection and 79% en bloc resection rates observed. Major bleeding was reported in 1 case (5%); there were no cases of perforation. With a median follow-up of 10 months, a single recurrence (5%) was observed. LIMITATIONS: A small series at a single center, with a short median follow-up time. CONCLUSION: We report the feasibility and effectiveness of gastric ESD in Europe. A further description of a Western series is expected, and guidelines for its dissemination are desirable to define the role of this technique in Western countries. PMID- 19185697 TI - Evaluation of hilar biliary strictures by using a newly developed forward-viewing therapeutic echoendoscope: preliminary results of an ongoing experience. AB - BACKGROUND: Obtaining a definitive tissue diagnosis in patients with hilar biliary strictures (HBS) is often difficult. OBJECTIVE: To describe our experience using a newly developed forward-viewing linear echoendoscope (FVL-EUS) with FNA as a primary diagnostic tool in patients with HBS. DESIGN: Case series. SETTING: A tertiary care, academic medical center. PATIENTS: Four patients with HBS who underwent the procedure. MAIN OUTCOME MEASUREMENTS: Performance of FNA with the FVL-EUS. RESULTS: Visualization and puncture of the primary lesion with a definitive tissue diagnosis was obtained in all of the 4 cases performed. Metastatic hilar cholangiocarcinoma and recurrent neuroendocrine tumor were diagnosed in 2 patients and followed by placement of a self-expandable metal stent, when possible. In the other 2 patients, a diagnosis of resectable hilar cholangiocarcinoma and poorly differentiated adenocarcinoma of unclear origin without evidence of vascular involvement was made, and plastic stents were placed before surgery; the first patient was found to have peritoneal metastases, and resection was aborted, and in the second patient, a gallbladder tumor was diagnosed in the surgical specimen. LIMITATION: The small number of patients. CONCLUSIONS: These preliminary data suggest that FVL-EUS used as a primary tool for the evaluation of patients with HBS may be of value and should be further explored in properly designed studies with a meaningful number of patients. PMID- 19185698 TI - Video capsule endoscopy in life-threatening GI hemorrhage after negative primary endoscopy (with video). AB - BACKGROUND: Video capsule endoscopy (VCE) continues to evolve as a key diagnostic tool. Traditionally VCE has been used to detect occult and obscure GI bleeding in adult patients. VCE has not been documented or accepted as an early diagnostic tool for acute life-threatening GI hemorrhage. OBJECTIVE: Our purpose was to demonstrate the use of VCE as an early diagnostic tool in acute life-threatening GI hemorrhage. DESIGN: Case series. PATIENTS: Patients with life-threatening GI hemorrhage. INTERVENTIONS: VCE after negative primary endoscopy. RESULTS: VCE allowed rapid diagnosis and reliable data before surgical intervention. Although proving to be a beneficial diagnostic tool for acute GI hemorrhage, VCE was not associated with increased morbidity or mortality rates. LIMITATIONS: This report only focuses on cases where VCE successfully led to a diagnosis. There is no prospective control group to which these patients can be compared. There were no other attempted acute VCE studies in patients with life-threatening bleeding during the time period of these case reports. CONCLUSIONS: The use of VCE is a simple and relatively safe diagnostic tool in the evaluation of continuing GI hemorrhaging in endoscopy-negative patients. The use of VCE can be considered as a another useful tool in the armamentarium of the endoscopist in the evaluation of GI bleeding. Prospective studies should be undertaken to determine the appropriate timing and clinical use in this group of patients. PMID- 19185700 TI - Larger short-axis length of lymph nodes predicts malignant involvement. PMID- 19185701 TI - Diagnostic bulb biopsies in celiac disease. PMID- 19185702 TI - Obtaining duodenal biopsy specimens for celiac disease: is site as important as number? PMID- 19185704 TI - Secrets of success in a difficult colonoscopy. PMID- 19185705 TI - The importance of "timing" when attempting to achieve cecal intubation of the "difficult colon". PMID- 19185706 TI - Warm water and oil for the difficult colon. PMID- 19185710 TI - Viability assays to monitor yeast autophagy. AB - In the yeast Saccharomyces cerevisiae, autophagy contributes to the sustaining of cell viability under starvation conditions, possibly through the supply of amino acids that is generated as a result of the degradation of cytosolic materials. Therefore, cellular viability is one of the best indexes for monitoring the completion of the entire autophagic process. In this chapter, several assays for monitoring yeast viability are presented. Along with the standard colony formation assay, assays using the dye phloxine B are introduced. PMID- 19185709 TI - Biochemical methods to monitor autophagy-related processes in yeast. AB - An increasing number of reports have elucidated the importance of macroautophagy in cell physiology and pathology. Macroautophagy occurs at a basal level and participates in the turnover of cytoplasmic constituents including long-lived proteins to maintain cellular homeostasis, but it also serves as an adaptive response to protect cells from various intra- or extracellular stresses. In addition, macroautophagy plays a role in development and aging and acts to protect against cancer, microbial invasion, and neurodegeneration. The machinery involved in carrying out this process, the autophagy-related (Atg) proteins were identified and characterized in various fungal systems, in particular because of the powerful tools available for genetic manipulation and the relative abundance of good biochemical assays in these model organisms. The analysis of these Atg proteins has allowed us to begin to understand the molecular mechanism of this process. Furthermore, many of the autophagy genes are functionally conserved in higher eukaryotes, including mammals, allowing the findings in fungi to be applied to other systems. Here, we discuss three biochemical methods to measure autophagy-related activities and to examine individual steps of the corresponding process. These methods rely on the detection of different modification states of certain marker proteins. Processing of the precursor form of the resident vacuolar hydrolase aminopeptidase I (Ape1) is applicable to fungi, whereas cleavage of the GFP-Atg8 and Pex14-GFP chimeras can be used in a wide array of systems. PMID- 19185711 TI - The quantitative Pho8Delta60 assay of nonspecific autophagy. AB - The measurement of autophagic flux is critical in understanding the regulation of autophagy. The Pho8Delta60 assay employs a very sensitive enzymatic assay that provides a high signal-to-noise ratio and allows for precise quantification of autophagic flow in yeast. Pho8, alkaline phosphatase, is a resident vacuolar enzyme that is delivered to the vacuole membrane through a portion of the secretory pathway. The assay utilizes a genetically engineered version of Pho8 that lacks the N-terminal transmembrane domain that allows for translocation into the endoplasmic reticulum. Accordingly, Pho8Delta60 remains in the cytosol and is delivered to the vacuole only through autophagy. Once in the vacuole lumen, the C terminal propeptide is proteolytically removed, which results in activation. Thus, the alkaline phosphatase activity reflects the amount of the cytosol delivered to the vacuole through nonspecific autophagy. PMID- 19185712 TI - Fluorescence microscopy-based assays for monitoring yeast Atg protein trafficking. AB - For several years, the yeast Saccharomyces cerevisiae has been the leading model organism for the study of autophagy. The amenability of this unicellular eukaryote to genetic and biochemical approaches has allowed the isolation and characterization of most of the genes specifically involved in autophagy, which are known as ATG (Reggiori, 2006; Reggiori and Klionsky, 2005). These pioneering studies have been of crucial relevance because most of the yeast ATG genes possess orthologs in all eukaryotic organisms. The experimental advantages, all the available reagents, and the established assays still maintain yeast in a prominent position in the study of autophagy and autophagy-related pathways. In this chapter, we describe fluorescent protein-based methodologies that permit one to readily assay the functionality of the autophagic pathway and to assess the trafficking of one of the key protein of this degradative process, Atg9. PMID- 19185713 TI - Measuring macroautophagy in S. cerevisiae: autophagic body accumulation and total protein turnover. AB - Macroautophagy has been implicated in various physiological functions and severe human diseases. Accordingly, there is a high interest in determining macroautophagy both qualitatively and quantitatively. In this chapter we discuss how macroautophagy can be followed morphologically in the yeast Saccharomyces cerevisiae using light microscopy. To quantitatively measure macroautophagy, we further present two protocols for the determination of total protein turnover. PMID- 19185714 TI - Aminopeptidase I enzymatic activity. AB - Aminopeptidase I is the cargo protein of the cytoplasm-to-vacuole targeting (Cvt), autophagy-like protein-targeting pathway of the yeast Saccharomyces cerevisiae, the nonclassical vacuolar biosynthetic transport route. The second enzyme following this route to the vacuole, alpha-mannosidase, is also transported by direct binding to the Atg19 receptor and to aminopeptidase I. Aminopeptidase I forms a homododecameric complex, which is synthesized and assembled in the cytoplasm, packed in double-membrane vesicles, and transported to the vacuole. Only the homododecameric complex of aminopeptidase I has exopeptidase activity directed against amino-terminal leucine residues. Enzymatic activity can be determined spectrofluorometrically in homogenates and semi quantitatively after nondenaturing gel electrophoresis and by yeast colony overlay assay. This chapter describes the methods to determine aminopeptidase I enzymatic activity used to follow complex assembly and vacuolar transport. PMID- 19185715 TI - Monitoring autophagy in yeast using FM 4-64 fluorescence. AB - The original observations and experiments dealing with autophagy were purely morphological in nature. Even though more and more molecular techniques have been introduced, experimenters are often asked to provide visual evidence of autophagic processes in order to back up data obtained via other means. In yeast as well, autophagosomes were initially defined morphologically and indirectly, by observing intravacuolar autophagic bodies that accumulate upon starvation. This can be achieved by electron microscopy, which affords very high resolution but is time consuming and costly, or by light microscopy, which is a relatively inaccurate method of scoring autophagy. A third alternative, which we present here, is to use the unique properties of the fluorescent dye FM 4-64 to follow the accumulation of autophagic bodies. PMID- 19185716 TI - Monitoring mitophagy in yeast. AB - Cellular degradative processes including proteasomal and vacuolar/lysosomal autophagic degradation, as well as the activity of proteases (both cytosolic and mitochondrial), provide for a continuous turnover of damaged and obsolete macromolecules and organelles. Mitochondria are essential for oxidative energy production in aerobic eukaryotic cells, where they are also required for multiple biosynthetic pathways to take place. Mitochondrial homeostasis also plays a crucial role in aging and programmed cell death, and recent data have suggested that mitochondrial degradation is a strictly regulated process. A recent study has shown that in yeast cells subjected to nitrogen starvation, degradation of mitochondria by autophagy occurs by both a selective process (termed mitophagy) and a nonselective process. This chapter provides an overview of the techniques that enable the study of mitophagy. Fluorescent proteins targeted to mitochondria can be used to follow mitochondrial sequestration within vacuoles. Degradation of mitochondria can be assayed using a mitochondrially targeted alkaline phosphatase (ALP) reporter test in which the delivery of mitochondrial N-terminal truncated Pho8Delta60 to the vacuole results from mitophagy. Degradation of mitochondrial proteins can also be followed by Western immunoblot analyses. Finally, electron microscopy observations permit the discrimination between selective mitophagy and nonselective mitochondrial degradation. PMID- 19185718 TI - Electron microscopy in yeast. AB - The membrane dynamics of autophagy in yeast were resolved mostly by using various electron microscopy (EM) methods combined with the cryofixation technique. In addition, the similarity of the dynamic process between autophagy and the cytoplasm-to-vacuole targeting (Cvt) pathway was first clarified through EM studies. In this chapter the application of several EM methods to detect the dynamic events involving cellular structures are described, and examples are provided of the typical images obtained in autophagy and the Cvt pathway. PMID- 19185717 TI - Monitoring organelle turnover in yeast using fluorescent protein tags. PMID- 19185719 TI - Cell-free reconstitution of microautophagy in yeast. AB - Microautophagy is the direct uptake of soluble or particulate cellular constituents into lysosomes. Here, I describe methods to reconstitute and study this process in vitro, using vacuoles (lysosomes) from the yeast Saccharomyces cerevisiae as model organelles. Protocols to grow the cells, isolate vacuoles from them, and to induce microautophagy of soluble tracers are presented. PMID- 19185720 TI - Autophagy in wine making. AB - Aging that involves contact with dying yeast cells is one of the differential processes between sparkling and still wine production. The release of the products of autolysis during this aging step is fundamental for the quality of sparkling wines made by the traditional method. These cells undergo an autolysis process characterized by self-digestion of yeast intracellular and cell-wall macromolecules, and the release of the degradation products to the wine. Autolysis is the source of several molecules responsible for the quality of sparkling wines, as well as still wines aged on lees (yeast cells). Autolysis is a slow process under sparkling wine production conditions, and there is interest, from the industrial side, in the design of strategies for rapid development of autolysis. Some years ago our research group hypothesized that, during the process of sparkling wine production, autophagy would take place. This had important implications for the design of genetic engineering strategies aimed to accelerate autolysis. The relationships between autolysis and autophagy are not completely elucidated, but in case autophagy preceded autolysis during the aging step of sparkling wine production, there were at least two possibilities for accelerating autolysis by targeting genes involved in autophagy. This chapter discusses methods to demonstrate the development of autophagy under enological conditions. This is accomplished by using either laboratory strains defective in autophagy and/or the Cvt pathway, in conditions that mimic sparkling wine production or industrial wine yeast strains under real sparkling wine production conditions. PMID- 19185721 TI - Purification and in vitro analysis of yeast vacuoles. AB - The purification of eukaryotic organelles is a prerequisite for the detailed analysis of protein sorting, localization and translocation, membrane fusion and vesicle budding. Yeast vacuoles receive cargo from the exocytic, endocytic, and autophagic pathways and hence represent an excellent model system for the study of organelle biogenesis and protein sorting. Yeast vacuoles undergo fission and fusion in vivo, events that can be monitored in vitro by an assay that employs purified vacuoles from two tester strains. Here, we describe the methodology of yeast vacuole purification, and provide protocols for the detailed analysis of the fusion reaction. We also include methods to analyze protein dynamics on yeast vacuoles and the controls required to ensure their reliability. PMID- 19185722 TI - Pexophagy in Hansenula polymorpha. AB - In the yeast Hansenula polymorpha the development and turnover of peroxisomes is readily achieved by manipulation of the cultivation conditions. The organelles massively develop when the cells are incubated in the presence of methanol as the sole source of carbon and energy. However, they are rapidly and selectively degraded when methanol-grown cells are placed at conditions of repression of methanol metabolism (e.g. in glucose or ethanol excess conditions) by a process termed macropexophagy. Degradation of peroxisomes is also observed when the cells are placed at nitrogen-depletion conditions (microautophagy). This contribution details the methodologies that are currently in use investigating macropexophagy and microautophagy in H. polymorpha. Emphasis is placed on various structural (fluorescence microscopy, electron microscopy) and biochemical (specific enzyme activity measurements, Western blotting) approaches. PMID- 19185723 TI - Pexophagy in Pichia pastoris. AB - The peroxisome is an organelle whose quantity is tightly regulated in response to changes in metabolic status, and much knowledge has been accumulated regarding its dynamics. The turnover of peroxisomes through autophagic pathways, termed pexophagy, has been especially studied in several methylotrophic yeast strains capable of growth on methanol as a sole carbon source, which led to the identification of factors involved in pexophagy (Dunn et al., 2005; Sakai et al., 2006). In the methylotrophic yeast Pichia pastoris, several types of membrane dynamics during pexophagy can be visualized simultaneously under live cell imaging. The decrease of abundant peroxisomal proteins in the cell lysate can be used as a convenient indicator of the completion of pexophagy. In combination, these methods provide basic information for further analysis of pexophagy at the molecular level. PMID- 19185724 TI - Methods of plate pexophagy monitoring and positive selection for ATG gene cloning in yeasts. AB - Methods for colony assay of peroxisomal oxidases in yeasts provide a convenient and fast approach for monitoring peroxisome status. They have been used in several laboratories for the isolation of yeast mutants deficient in selective autophagic peroxisome degradation (pexophagy), catabolite repression of peroxisomal enzymes or mutants deficient in oxidases themselves. In this chapter, protocols for monitoring peroxisomal alcohol oxidase and amine oxidase directly in yeast colonies and examples of their application for mutant isolation are described. These methods were successfully utilized in several methylotrophic yeasts and the alkane-utilizing yeast Yarrowia lipolytica. PMID- 19185725 TI - Autophagy in the filamentous fungus Aspergillus fumigatus. AB - Many breakthroughs in our understanding of the function and molecular basis of autophagy have been achieved in mammalian and yeast systems. However, we still know very little about the contribution of autophagy to the biology of filamentous fungi. A comparative analysis of autophagy between genera will expand our knowledge of the autophagy machinery and has the potential to identify novel functions that are relevant to multiple biological systems. This chapter will discuss methods that have been employed for studying autophagy in the opportunistic mold pathogen Aspergillus fumigatus. Understanding how autophagy influences the growth of this important human pathogen could lead to the development of novel antifungal drugs that restrict the growth of the fungus by manipulating the autophagy pathway. PMID- 19185726 TI - Monitoring autophagy in the filamentous fungus Podospora anserina. AB - Autophagy has been monitored in the filamentous fungus Podospora anserina using electron, light, and fluorescence microscopy. In this organism autophagy can be induced either by starvation or rapamycin treatment or by het gene incompatibility. Incompatible HET products signal a cell death reaction referred to as cell death by incompatibility. In het-R het-V strain bearing the two incompatible het-R and het-V genes, cell death is induced by a simple shift in growth temperature, as incompatibility is thermosensitive. In this strain large autophagosomes are formed as revealed by electron microscopy or using the GFP PaATG8 marker. This strain constitutes an alternative model to study autophagy. Analysis of the three autophagy mutants, DeltaPaATG1, DeltaPaATG8, and DeltapspA, reveals that autophagy is essential for aerial hyphae and female organ differentiation and involved in spore germination. During the incompatibility reaction, autophagy might protect cells from cell death as suggested by accelerated cell death observed in autophagy mutants. PMID- 19185727 TI - Monitoring autophagy in Magnaporthe oryzae. AB - Autophagy is a ubiquitous degradative pathway for the bulk degradation of eukaryotic macromolecules and organelles in eukaryotic cells (Klionsky, 2005; Levine and Klionsky, 2004). Previously, the role of autophagy in turgor generation in plant pathogenic fungi was unknown. Currently, autophagy is confirmed as an important pathway for turgor accumulation in the appressorium (the tips of the invasive hyphae; Liu et al., 2007b) using a technique of targeted gene replacement, deleting the genes that code for Magnaporthe oryzae homologs of yeast autophagy-related (ATG) genes ATG2, ATG4, ATG5, ATG8, ATG9, and ATG18 (Liu et al., 2007a). All of these null mutants fail to breach the cuticle of the host. This chapter will first look at some methodologies to analyze the functions of autophagy-related gene products at the biological, cellular, and molecular level in this model plant pathogenic fungi, and then provide some research evidence of the role of autophagy in the promotion of the formation of the infection structure and pathogenicity to point out some significant areas for further research in this field. PMID- 19185728 TI - Methods for functional analysis of macroautophagy in filamentous fungi. AB - Autophagy is a bulk degradative process responsible for the turnover of membranes, organelles, and proteins in eukaryotic cells. Genetic and molecular regulation of autophagy has been independently elucidated in budding yeast and mammalian cells. In filamentous fungi, autophagy is required for several important physiological functions, such as asexual and sexual differentiation, pathogenic development, starvation stress and programmed cell death during heteroincompatibility. Here, we detail biochemical and microscopy methods useful for measuring the rate of induction of autophagy in filamentous fungi, and we summarize the methods that have been routinely used for monitoring macroautophagy in both yeast and filamentous fungi. The role of autophagy in carbohydrate catabolism and cell survival is discussed along with the specific functions of macroautophagy in fungal development and pathogenesis. PMID- 19185729 TI - Autophagy in Candida albicans. AB - Candida albicans is considered a commensal organism of humans, colonizing the oral cavity, gastrointestinal, and reproductive tracts. However, when host defenses are compromised C. albicans can transform into a tissue invasive pathogen. Infections fall into two broad categories, those of mucosal tissue and serious disseminated disease, which involves transport through the bloodstream and invasion of the deeper organs. The ability of C. albicans to colonize diverse host microenvironments and switch from benign commensal to invasive pathogen suggests that C. albicans is able to undergo rapid and highly specialized adaptive responses in vivo. To date the role played by autophagy in facilitating asymptomatic host colonization, persistence, and transition of C. albicans into its pathogenic form have not been fully explored. Also the therapeutic potential of manipulating autophagic degradation within an invading fungus is yet to be established. In this chapter we describe several methods that have been adapted to detect autophagy within the opportunistic pathogen C. albicans. PMID- 19185730 TI - Analysis of autophagy during infections of Cryptococcus neoformans. AB - Cryptococcus neoformans is a yeastlike fungus that causes a lethal meningoencephalitis in a broad spectrum of immunocompromised patients and has become the most common cause of meningitis due to AIDS-related infections in Africa. Key to the development of new agents to control and prevent this infection is the identification of cellular mechanisms required for pathogenesis. Survival of the fungus within the hostile and nutrient-deprived environments of the host has recently been shown to depend on the induction of autophagy, whereby the cell recycles nutrients by slowly digesting itself in a regulated fashion. Further study of the role of autophagy during infection by C. neoformans requires the use of markers of autophagy that are specially adapted to the fungus within the mammalian host. PMID- 19185731 TI - Autophagy and autophagic cell death in Dictyostelium. AB - Autophagic cell death can be conveniently studied in Dictyostelium discoideum, an exceptionally favorable model not only because of its well-known genetic and experimental advantages but also because in Dictyostelium there is no apoptosis machinery that could interfere with nonapoptotic cell death. Moreover, autophagic cell death in Dictyostelium can be dissociated into a starvation-induced sensitization stage, during which autophagy is induced, and a death induction stage. We show here how to demonstrate, assess and analyze this autophagic cell death. This can be studied in vivo during the development of Dictyostelium, and in vitro, using modifications of the monolayer technique of Rob Kay et al. Methods to follow this autophagic cell death qualitatively and quantitatively are reported. PMID- 19185732 TI - Analysis of autophagy in the enteric protozoan parasite Entamoeba. AB - Entamoeba histolytica is the enteric protozoan parasite that causes human amoebiasis. We have previously shown that autophagy is involved in proliferation and differentiation in the related species Entamoeba invadens, which infects reptiles and develops similar clinical manifestations. Because this group of protists possesses only a limited set of genes known to participate in autophagy in other eukaryotes, it potentially represents a useful model for studying the core system of autophagy and provides tools to elucidate the evolution of eukaryotes and their organelles. Here we describe the methods to study autophagy in Entamoeba. PMID- 19185733 TI - Kinetoplastida: model organisms for simple autophagic pathways? AB - Phylogenetic analyses based on defined proteins or different RNA species have revealed that the order kinetoplastida belongs to the early-branching eukaryotes and may thus contain organisms in which complex cellular events are easier to analyze. This view was further supported by results from a bioinformatic survey that suggested that nearly half of the autophagy-related proteins existent in yeast are missing in trypanosomatids. On the other hand, these organisms have evolved a highly sophisticated machinery to escape from the different host immune response strategies and have learned to cope with extremely variable environmental conditions by morphological and functional reorganization of the cell. For both the stress response and the differentiation processes, autophagy seems to be an indispensable prerequisite. So far autophagy has not been systematically investigated in trypanosomatids. Here we present technical information on how to handle the different parasites belonging to this order and give an overview of the current status of autophagy research in these organisms. PMID- 19185734 TI - Methods to investigate autophagy during starvation and regeneration in hydra. AB - In hydra, the regulation of the balance between cell death and cell survival is essential to maintain homeostasis across the animal and promote animal survival during starvation. Moreover, this balance also appears to play a key role during regeneration of the apical head region. The recent finding that autophagy is a crucial component of this balance strengthens the value of the Hydra model system to analyze the implications of autophagy in starvation, stress response and regeneration. We describe here how we adapted to Hydra some established tools to monitor steady-state autophagy. The ATG8/LC3 marker used in biochemical and immunohistochemical analyses showed a significant increase in autophagosome formation in digestive cells after 11 days of starvation. Moreover, the maceration procedure that keeps intact the morphology of the various cell types allows the quantification of the autophagosomes and autolysosomes in any cell type, thanks to the detection of the MitoFluor or LysoTracker dyes combined with the anti-LC3, anti-LBPA, and/or anti-RSK (ribosomal S6 kinase) immunostaining. The classical activator (rapamycin) and inhibitors (wortmannin, bafilomycin A(1)) of autophagy also appear to be valuable tools to modulate autophagy in hydra, as daily-fed and starved hydra display slightly different responses. Finally, we show that the genetic circuitry underlying autophagy can be qualitatively and quantitatively tested through RNA interference in hydra repeatedly exposed to double-stranded RNAs. PMID- 19185735 TI - Autophagy in freshwater planarians. AB - Planarians provide a new and emergent in vivo model organism to study autophagy. On the whole, maintaining the normal homeostatic balance in planarians requires continuous dynamic adjustment of many processes, including proliferation, apoptosis, differentiation, and autophagy. This makes them very different from other models where autophagy only occurs at very specific times and/or in very specific organs. This chapter aims to offer a general vision of planarians as a model organism, placing more emphasis on those characteristics related to autophagy and describing how autophagy fits into the processes of body remodeling during regeneration and starvation. We also define exactly what is known about autophagy in these organisms and we discuss the techniques available to study the relevant processes, as well as the techniques that are currently being developed. As such, this chapter will serve as a compilation of the techniques available to investigate autophagy in planarians. PMID- 19185736 TI - Qualitative and quantitative characterization of autophagy in Caenorhabditis elegans by electron microscopy. AB - Caenorhabditis elegans has been introduced relatively late into the field of autophagy with no previous results by classical methods. Therefore, it has to be studied in parallel with both traditional electron microscopy and modern molecular approaches. In general, correct identification of autophagic elements by electron microscopy is indispensable to establish a firm basis for our understanding of the process. The principles and the method for identification, applied also for C. elegans, are summarized first in this article, to facilitate their utilization both for further studies and the analysis of new cell types and to support researchers new to electron microscopy techniques. Studying autophagy in the worm by electron microscopy has required the development of special handling and sampling techniques in addition to overcoming the general technical difficulties due to the nature of C. elegans samples. These are described in detail, together with some initial qualitative and quantitative results obtained by them. The feasibility of the presented method is supported by data which show that in continuously fed worms the autophagic compartment is in the lower range of the 10(-2)% order of magnitude of the cytoplasmic volume, while immediately after molting or upon starvation in the second larval period, usually more than a 10-fold increase can be measured. In dauer larvae, individual variation of the autophagic compartment is very high. The predauer stage in daf-2 mutants does not seem to show significant constitutive autophagic activity. Some autophagy-related gene mutants show characteristic ultrastuctural features, such as autophagosomes with membrane abnormalities (unc-51/Atg1) or the hypertrophy of multivesicular bodies (let-512/Vps34, bec-1/Atg6). PMID- 19185737 TI - Monitoring the role of autophagy in C. elegans aging. AB - Autophagy plays crucial roles in many biological processes, and recent research points to a possibly conserved role for autophagy in the process of organismal aging. Experiments in the nematode C. elegans suggest that autophagy may be required specifically for longevity pathways that are regulated by environmental signals. Known longevity genes can be assigned to four major longevity pathways/processes: insulin/IGF-1 signaling, dietary restriction, protein translation, and mitochondrial respiration. Of these, reduced insulin/IGF-1 signaling and dietary restriction, but not protein translation inhibition, appear to rely on autophagy to increase life span. Multiple experimental approaches have been used to study autophagy in the context of aging in C. elegans. This chapter describes techniques used to address the link between aging and autophagy in C. elegans. Specifically, we summarize how to examine organismal life span in various longevity mutants and how to visually detect autophagy and auto-lysosomal formation in C. elegans. PMID- 19185738 TI - Autophagy in Caenorhabditis elegans. AB - Autophagy (cellular self-eating) is a highly regulated, lysosome-mediated catabolic process of eukaryotic cells to segregate by a special membrane and subsequently degrade their own constituents during development or starvation. Electron microscopy analysis reveals autophagic elements in various cell types of the nematode Caenorhabditis elegans, whose genome contains counterparts of several yeast genes involved in autophagy. Genetic manipulation inactivating autophagy-related genes in C. elegans causes defects in development, affects dauer larval morphogenesis, accelerates aging thereby shortening life span, reduces cell size, decreases survival during starvation, promotes apoptotic cell death, and protects neurons from undergoing hyperactive ion channel- or neurotoxin-induced degeneration. These results implicate autophagy in various developmental and cellular functions such as reproductive growth, aging, and cell growth, as well as cell survival and loss. This chapter discusses methods of inactivating C. elegans autophagy genes by RNA interference, testing the resistance of autophagy-deficient nematodes to starvation-induced stress, handling mutants carrying a deletion in the autophagy pathway, and monitoring autophagic activity by using LysoTracker Red dye or reporters labeled with green fluorescent protein. Such methods may be adaptable to identify additional roles of autophagy in development and cellular function, and may also help to detect the intracellular accumulation of autophagy proteins and monitor autophagosome formation. PMID- 19185739 TI - Chimeric fluorescent fusion proteins to monitor autophagy in plants. AB - Autophagy is induced under nutrient-deficient conditions in both growing tobacco BY-2 cultured cells as well as Arabidopsis and others intact plants. The fluorescent protein-tagged structural protein for autophagosomes, the Atg8 protein, allows nondestructive detection of autophagy induction in plant cells and tissues by fluorescence microscopy. Using this technique, the general operation of autophagy in growing root cells has been observed. A synthetic cargo protein for autophagy consisting of cytochrome b5 and the red fluorescence protein, DsRed, allows for the quantitative assay of autophagy in tobacco cells. This chapter describes methods for detecting autophagy in these plant cells using fluorescent protein fusions in situ with light microscopy, as well as quantification of autophagy. PMID- 19185740 TI - Use of protease inhibitors for detecting autophagy in plants. AB - In cultured tobacco (BY-2) cells, autophagy seems to be induced under nutrient starvation conditions, whereas in root cells from Arabidopsis and barley, it occurs constitutively though is activated under nutrient starvation conditions. In both cases, protease inhibitors such as E-64, E-64c, antipain, and leupeptin block autophagy at the step of degradation of the cytoplasm enclosed in lysosomes/vacuoles, and cause the accumulation of autolysosomes (lysosomes containing parts of the cytoplasm) and/or of many cytoplasmic inclusions in the central vacuoles. Both types of autophagy are inhibited by 3-methyladenine, which is known as a potent inhibitor of autophagy in mammalian cells. Thus, using protease inhibitors and 3-methyladenine provides us with a method useful for analyzing autophagy in plant cells. This chapter describes protocols for detecting autophagic compartments in BY-2 cells and in the root-tip cells of Arabidopsis and barley by microscopy. PMID- 19185741 TI - Lysosomes and autophagy in aquatic animals. AB - The lysosomal-autophagic system appears to be a common target for many environmental pollutants, as lysosomes accumulate many toxic metals and organic xenobiotics, which perturb normal function and damage the lysosomal membrane. In fact, autophagic reactions frequently involving reduced lysosomal membrane integrity or stability appear to be effective generic indicators of cellular well being in eukaryotes: in social amoebae (slime mold), mollusks and fish, autophagy/membrane destabilization is correlated with many stress and toxicological responses and pathological reactions. Prognostic use of adverse lysosomal and autophagic reactions to environmental pollutants can be used for predicting cellular dysfunction and health in aquatic animals, such as shellfish and fish, which are extensively used as sensitive bioindicators in monitoring ecosystem health; and also represent a significant food resource for at least 20% of the global human population. Explanatory frameworks for prediction of pollutant impact on health have been derived encompassing a conceptual mechanistic model linking lysosomal damage and autophagic dysfunction with injury to cells and tissues. Methods are described for tracking in vivo autophagy of fluorescently labeled cytoplasmic proteins, measuring degradation of radiolabeled intracellular proteins and morphometric measurement of lysosomal/cytoplasmic volume ratio. Additional methods for the determination of lysosomal membrane stability in lower animals are also described, which can be applied to frozen tissue sections, protozoans and isolated cells in vivo. Experimental and simulated results have also indicated that nutritional deprivation (analogous in marine mussels to caloric restriction)-induced autophagy has a protective function against toxic effects mediated by reactive oxygen species (ROS). Finally, coupled measurement of lysosomal-autophagic reactions and simulation modelling is proposed as a practical toolbox for predicting toxic environmental risk. PMID- 19185742 TI - Autophagy in ticks. AB - The generation time of ticks is estimated at several years and most ticks spend more than 95% of their life off the host. They seem to have a unique strategy to endure the off-host state for a long period. We focused on autophagy that is induced by starvation and is essential for extension of the life span in model organisms. Autophagy may occur in ticks that can survive extended periods of starvation. Although little research has been done on autophagy in ticks, recently, we showed the existence of an ATG gene homolog, HlATG12, in the 3-host tick Haemaphysalis longicornis. We have also examined the expression patterns of HlATG12, from nymphal to adult stages of this tick and revealed the localization of the HlAtg12, protein within midgut epithelial cells of unfed adult ticks. However, autophagy in ticks is a new field, so methods for monitoring this phenomenon in ticks are still to be established. This chapter discusses protocols for the detection of HlATG12, gene/HlAtg12, protein and the observation of the midgut epithelial cells using an electron microscope during the nonfeeding period of H. longicornis ticks. These methods can be adapted and modified for the study autophagy in other hard ticks. PMID- 19185743 TI - Quantitative analysis of autophagic activity in Drosophila neural tissues by measuring the turnover rates of pathway substrates. AB - The process of macroautophagy occurs in most eukaryotic cells and serves as the main recycling mechanism for the elimination of excess cytoplasmic components. The pathway is upregulated under a wide range of stress-related conditions and basal levels of autophagy are critical for the clearance of age-associated cellular damage, which can accumulate in long-lived, nondividing cells such as neurons. Traditionally, activation of autophagy has been measured by the microscopic observation of newly formed autophagosomes or by monitoring the further modification of the LC3-I protein to the LC3-II isoform by Western blot analysis. However, using these methods to quantitatively determine autophagic activity that occurs in complex tissues over an entire life span has been a technical challenge and difficult to consistently reproduce. We have shown that Western analysis of protein substrates normally cleared by the pathway can be used to make quantitative estimates of autophagy occurring in tissues such as the adult Drosophila nervous system. By examining the profile of insoluble ubiquitinated proteins (aggregated proteins) we have found that an age-dependent decline in pathway flux or genetic defects in critical autophagic genes can result in the concomitant buildup of substrates that are normally targeted by autophagy to the lysosome. Further, we have found that increasing Atg81a expression (a key rate-limiting component of the pathway) during the time in which autophagy is normally suppressed prevents the age-dependent accumulation of insoluble ubiquitinated proteins in neurons. This technique, as well as the detection of proteins damaged by reactive carbonyl groups, can also be used to measure autophagic activity in both normal and genetically altered flies during the aging process or following their acute exposure to oxidants. PMID- 19185744 TI - Genetic manipulation and monitoring of autophagy in Drosophila. AB - Drosophila melanogaster provides a model system useful for many aspects of the study of autophagy in vivo. These include testing and validation of genes potentially involved in autophagy, discovery of novel genes through genetic screening for mutations that affect autophagy, and analysis of potential roles of autophagy in specific developmental or physiological processes. In recent years, a number of techniques and transgenic and mutant fly strains have been developed to facilitate autophagy analysis in this system. Here, protocols are described for activating or inhibiting autophagy in Drosophila, and for examining the progression of autophagy in vivo through imaging-based assays. The goal of this chapter is to provide a resource both for autophagy investigators with limited familiarity with fly genetics, as well as for experienced Drosophila biologists who wish to test for connections between autophagy and a given gene, pathway or process. PMID- 19185745 TI - Monitoring autophagy in insect eggs. AB - Oogenesis is a fundamental physiological process in insects. Successful oogenesis is critical for evolutionary success by transferring genetic information to the next generation. This is achieved by the normal maturation of the egg chamber (egg), which is accomplished through cell death of the cells that accompany the oocyte. Recent studies demonstrate that autophagy contributes to this cell death process. Hence, comprehension of the mechanisms that implicates autophagy during cell death in insect eggs is very important. Herein, we describe some experimental approaches that can be used to monitor autophagy in insect eggs. PMID- 19185746 TI - In vitro methods to monitor autophagy in Lepidoptera. AB - Autophagy is attracting growing interest, especially in relation to increasing evidence of the importance of autophagic processes in animal development, as well as in human cancer progression. In holometabolous insects (i.e., that undergo four distinct life cycle stages, including embryo, larva, pupa and imago), such as flies, butterflies, bees and beetles, autophagy has been found to play a fundamental role in metamorphosis, and given the high degree of conservation of the genes and the basic mechanisms of autophagy, attention to these relatively simple models has increased significantly. Together with Drosophila, Lepidoptera larvae are among the most common invertebrate models in studies concerning the protective action of starvation-induced autophagy or the possible role of autophagy as a programmed cell death process. In this chapter, we provide experimental methods developed for, or applicable to, the study of the autophagic process in the IPLB-LdFB cell line derived from the fat body of the caterpillar of the gypsy moth, Lymantria dispar. PMID- 19185759 TI - Re: Somani BK et al.: Successful management of "overlooked" ureteral stent in transplant kidney (Urology 2008;72:1012). PMID- 19185761 TI - Re: Chien-Hui O, et al.: En bloc stapling of renal hilum during hand-assisted retroperitoneoscopic nephroureterectomy in dialysis patients (Urology. 2008;72:589-592). PMID- 19185763 TI - Re: Berger AD, et al.: Transperitoneal laparoscopic radical nephrectomy for large (>7 cm) renal masses (Urology 2008;71:421-424). PMID- 19185764 TI - Re: Henderson A et al: Hand-assisted laparoscopic nephrectomy for renal cell cancer with renal vein tumor thrombus (Urology 2008;72:268-272). PMID- 19185765 TI - Re: Harris SE, Guralnick ML, O'Connor RC: Urethral erosion of transobturator male sling. (Urology 2009;73:443). PMID- 19185768 TI - Re: Agarwal a et al.: The safety and efficacy of different doses of bacillus Calmette Guerin in superficial bladder transitional cell carcinoma (Urology 2007;70:1075-1078). PMID- 19185769 TI - Let's not have to hear it through the grapevine. PMID- 19185770 TI - Surface EMG application in clinical biomechanics. Editorial. PMID- 19185771 TI - Hypercaloric diets differing in fat composition have similar effects on serum leptin and weight gain in female subjects with anorexia nervosa. AB - Weight regain in subjects with anorexia nervosa is associated with an increase in serum leptin concentrations that is hypothesized to impair full weight restoration. As diets rich in n-3 polyunsaturated fatty acids (PUFA) have been described to lower serum leptin concentrations, we tested the hypothesis that consumption of a hypercaloric diet rich in n-3 PUFA is associated with an attenuated increase in serum leptin and a higher efficiency of body weight gain in subjects with anorexia nervosa. Twenty-five female subjects with anorexia nervosa were enrolled into this controlled dietary intervention study. Four subjects discontinued therapy or participation in the study prematurely, and six were excluded. 15 subjects completed the study. Subjects consumed hypercaloric diets rich in either saturated fatty acids (SFA, n = 8) or n-3 PUFA (n = 7) for 5 weeks. Primary endpoints were the change in serum leptin concentrations and body weight gain relative to energy consumed. Serum leptin concentrations increased distinctly throughout the study (P < .001), and to a similar extend in both groups [+2.9 (SD 2.4) vs. +2.8 (SD 3.4) ng/mL in the SFA- and n-3 PUFA group, respectively; P = .487]. The efficiency of body weight gain also did not differ significantly between groups, with a body weight gain of 63.1 (SD 12.4) vs. 79.2 (SD 26.0) g per 4.2 MJ (1000 kcal) consumed in the SFA- and n-3 PUFA group, respectively (P = .132). Hypercaloric diets rich in either SFA or n-3 PUFA do not differ in their effects on serum leptin concentrations and the efficiency of body weight gain in female subjects with anorexia nervosa. PMID- 19185772 TI - Low-sodium Dietary Approaches to Stop Hypertension-type diet including lean red meat lowers blood pressure in postmenopausal women. AB - Low-sodium Dietary Approaches to Stop Hypertension (DASH) diets are base producing but restrict red meat without clear justification. We hypothesized that a vitality diet (VD), a low-sodium DASH-type diet with a low dietary acid load containing 6 servings of 100 g cooked lean red meat per week, would be more effective in reducing blood pressure (BP) compared with a higher acid load reference healthy diet (RHD) based on general dietary guidelines to reduce fat intake and increase intake of breads and cereals. A randomized, parallel dietary intervention study was conducted to compare the BP-lowering effect of these 2 diets in postmenopausal women with high/normal BP. Women were randomly assigned to follow either VD or RHD for 14 weeks. Home BP was measured daily with an automated BP monitor under standard conditions. Of 111 women commencing the study, 95 completed (46 VD, 49 RHD). Systolic BP (SBP) throughout the intervention was lower in the VD group compared to the RHD group (repeated measures analysis of variance time by diet, P = .04), such that at the end of the study, the VD had a fall of SBP by 5.6 +/- 1.3 mm Hg (mean +/- SEM) compared with a fall of 2.7 +/- 1.0 mm Hg in the RHD (group difference, P = .08). When only those taking antihypertensive medications were assessed, the VD (n = 17) had a significant fall of 6.5 +/- 2.5 mm Hg SBP (P = .02) and 4.6 +/- 1.4 mm Hg diastolic BP (P = .005) after 14 weeks, and their BP was lower than that of the RHD group (n = 18) throughout the study (P < .05). We concluded that a low-sodium DASH diet with a low dietary acid load, which also included lean red meat on most days of the week, was effective in reducing BP in older women, particularly in those taking antihypertensive medications. PMID- 19185773 TI - Lycium barbarum (goji) juice improves in vivo antioxidant biomarkers in serum of healthy adults. AB - Although Lycium barbarum (goji) and active compounds, Lycium barbarum polysaccharides (LBP), have a high in vitro antioxidant score as determined by simple chemical reaction methods, their in vivo antioxidant effects in humans have not been extensively examined. After our earlier report that an LBP standardized Lycium barbarum preparation (GoChi) helps prevent oxidant stress related conditions in humans, our present study examined the hypothesis that the antioxidant effects of GoChi result from its ability to enhance endogenous antioxidant factors. We investigated the effects of GoChi in a 30-day randomized, double-blind, placebo-controlled clinical study. The study population included 50 Chinese healthy adults aged 55 to 72 years. In vivo antioxidant markers, consisting of serum levels of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and lipid peroxidation (indicated by decreased levels of malondialdehyde, MDA) were examined preintervention and postintervention with GoChi or placebo (120 mL/d). In the GoChi group, antioxidant markers significantly increased by 8.4% for SOD and 9.9% for GSH-Px between the preintervention and postintervention measurements, whereas MDA were significantly decreased by 8.7%. In addition, the SOD, GSH-Px, and MDA levels in the GoChi group were significantly different from those in the placebo group at the postintervention time point, with increases of 8.1% and 9.0% and a decrease of 6.0%, respectively. No significant differences were detected between the preintervention and postintervention time points in the placebo group. These results indicate that GoChi increased antioxidant efficacies in humans by stimulating endogenous factors and suggest that continued use beyond 30 days might help prevent or reduce free radical-related conditions. PMID- 19185775 TI - Protein-energy malnutrition contributes to increased structural chromosomal alteration frequencies in Argentinean children. AB - The relationship between protein-energy malnutrition and genetic damage has been studied in human beings and laboratory animals, but results are still conflicting. The aim of the present study was to assess the induction of structural chromosomal aberrations in peripheral blood lymphocytes of children with primary protein-energy malnutrition. A case-control study was performed. Samples were obtained from 25 primary malnourished infants (mean age, 22 months; range, 1-66 months). The control group consisted of 25 eutrophic children from the same population who were matched 1:1 by age and sex. Anthropometric and clinic evaluations were performed to assess nutritional condition. Before blood collection, we interviewed each individual's parent to complete a semi-structural survey specifying age, dietary habits, viral or bacterial diseases; previous exposure to diagnostic x-rays; and use of therapeutic drugs. After 48 hours, 100 cultured lymphocytes were analyzed per patient. Statistical analysis was performed using the Epi Dat 3.0 program (P < or = .05). The chromosomal aberration frequency was nearly 7 times higher in malnourished infants than in controls (14.61% vs 2.2%, respectively). This difference was statistically significant (P < .001) and may be explained by the occurrence of achromatic lesions, breaks, and telomeric associations. DNA damage could be attributed to several factors: severe deficiency of essential nutrients (ie zinc, iron, and vitamin A) required in the synthesis of DNA maintenance factors; deterioration of repair mechanisms allowing the persistence of an unusually high number of structural chromosomal aberrations; and/or the absence of specific factors needed to protect the cell against oxidative DNA damage. PMID- 19185774 TI - A low pretransplant peripheral blood mononuclear cell complex I activity predicts metabolic disturbances and inability to regain fat free mass in cirrhotic patients undergoing liver transplantation. AB - Liver cirrhosis is associated with malnutrition and often, after liver transplantation, with the development of obesity and the inability to gain lean body mass. We have previously shown that peripheral blood mononuclear cell (PBMNC) complex I activity could be an appropriate marker for nutritional assessment. In this context, we hypothesized that a low pretransplant PBMNC complex I activity may predict a poor nutritional status in cirrhotic patients undergoing liver transplantation. Fifteen cirrhotic patients (CP) (8 men and 7 women) were recruited and investigated before and 4 months after liver transplantation. Body weight, body composition by DEXA, anthropometric measures (triceps skinfold thickness and midarm muscle circumference), resting energy expenditure, respiratory quotient and PBMNC complex I activity were measured on both time points. Patients were divided into 2 groups depending on their pretransplant PBMNC complex I activity (low vs high complex I activity [CP(low CI) vs CP(high CI)]), using as an arbitrary cutoff value-the mean complex I activity observed in age-matched healthy controls. Before transplantation, the CP(low CI) group who showed a lower complex I activity (2.11 +/- 0.53 vs 4.54 +/- 0.98 nmol/min per milligram of protein, P < .01) was significantly younger (44 +/ 9 vs 62 +/- 8 years old, P < .01); no differences were observed for any other nutritional parameters when compared to the CP(high CI) group. After transplantation, only the CP(low CI) group demonstrated a significant increase of complex I activity (+77%, P < .01), respiratory quotient (+10.5%, P < .02), triceps skinfold thickness (+126%, P < .005), and a significant decrease of fat free mass (-8%, P < .01). In summary, our findings indicate that a low pretransplant PBMNC complex I activity in cirrhotic patients could be a useful marker of poor nutritional status despite the lack of traditional indicators of malnutrition by predicting metabolic disturbances and an inability to regain fat free mass after liver transplantation. PMID- 19185776 TI - Dietary selenium intake is negatively associated with serum sialic acid and metabolic syndrome features in healthy young adults. AB - Low-grade and chronic inflammation related to excessive body weight can increase the risk for type 2 diabetes and cardiovascular disease, whereas the intake of antioxidant nutrients appears to produce anti-inflammatory effects. The purpose of this observational study was to assess the potential relationships between serum SA levels, metabolic syndrome features, and dietary selenium intake to test the hypothesis that this antioxidant micronutrient may also have anti inflammatory properties in healthy young adults. Forty-three healthy participants with a mean age of 18.0 +/- 0.93 years and a mean body mass index of 22.2 +/- 2.7 kg/m(2) were enrolled. Anthropometric, body composition, and blood pressure determinations were measured as well as serum lipid profile, glucose, insulin, and SA concentrations. Nutritional intake was estimated by a computerized, validated semiquantitative food frequency questionnaire. The findings included a positive correlation between SA and triacylglycerol levels (r = 0.317, P = .038) and a trend to significance with the homeostatic model assessment of insulin resistance index (r = 0.297, P = .053). Moreover, subjects with higher dietary selenium intake showed statistically lower SA levels compared with subjects with lower dietary selenium intake (1.8 +/- 0.4 vs 2.1 +/- 0.4 mmol/L, P = .037), while dietary selenium negatively correlated with SA (r = -0.331, P = .030) and triacylglycerol levels (r = -0.312, P = .041). It can be concluded that a relationship of SA, an inflammatory marker, with metabolic syndrome features such as lipid profile impairment and insulin resistance has been envisaged. In addition, we report (apparently for the first time) a negative association between SA and selenium intake, a recognized antioxidant trace element, in healthy young subjects, reinforcing the view of selenium as a potential anti inflammatory nutrient. PMID- 19185777 TI - In vitro intestinal glucose uptake is inhibited by galactomannan from Canadian fenugreek seed (Trigonella foenum graecum L) in genetically lean and obese rats. AB - Galactomannan, a soluble fiber, has been reported to reduce postprandial blood glucose response. Using this fiber, extracted from Canadian-grown fenugreek seeds (Trigonella foenum graecum L), we conducted an in vitro study to determine if galactomannan affects intestinal glucose uptake in genetically determined lean and obese rats. The segments of jejunum and ileum from these animals were incubated with labeled glucose (2 or 32 mmol/L) in the presence of different concentrations of galactomannan ranging from 0.1% to 0.5% (wt/wt). The uptake of low or high concentration of glucose was significantly and progressively reduced by increasing concentrations of galactomannan in both lean and obese rats. No significant difference was observed in the uptake of glucose between the 2 groups. The viscosity of various concentrations of galactomannan solutions was determined after stirring for 60 minutes at a temperature-controlled (37 degrees C) fixed sheer rate of 1.29 (1/s). The inhibitory effect of galactomannan on glucose uptake was found to be in parallel with the degree of viscosity of the fiber solutions. These results suggest that the galactomannan, because of its viscous property, has the potential to reduce intestinal absorption of low or high concentrations of glucose and hence for the benefit of blood glucose management. PMID- 19185778 TI - A mixture of Salacia reticulata (Kotala himbutu) aqueous extract and cyclodextrin reduces body weight gain, visceral fat accumulation, and total cholesterol and insulin increases in male Wistar fatty rats. AB - This study examined the effects of a mixture of an aqueous extract of Salacia reticulata (Kotala himbutu) and cyclodextrin (SRCD) on various metabolic parameters and cecal fermentation in obese fa/fa male Wistar fatty rats, a model of type 2 diabetes mellitus. Wistar fatty rats were fed 0% (control group) or 0.2% SRCD-supplemented diets and weighed weekly. The plasma glucose, triacylglycerol, total cholesterol, insulin, and adiponectin concentrations were measured at weeks 0, 2, 4, and 5. SRCD supplementation suppressed the time dependent increase in the plasma total cholesterol and insulin concentrations. After 6 weeks of a 0.2% SRCD-supplemented diet, the body weight gain, food intake, visceral fat mass, liver mass, and liver triacylglycerol content of the rats were significantly lower, whereas the plasma adiponectin concentrations were significantly higher than those of the control group. SRCD supplementation had no significant effect on plasma glucose and triacylglycerol concentrations. SRCD supplementation significantly increased cecum mass, whereas it significantly decreased the cecal butyrate and short-chain fatty acid (sum of the acetate, butyrate, and propionate) concentrations. All of the rats were subjected to an oral glucose tolerance test at the beginning of week 6. The area under the curve for insulin was significantly smaller with SRCD supplementation and showed no change in glucose tolerance compared to that of the control group. These results suggest that bioactive compounds in SRCD may suppress the development of type 2 diabetes mellitus by influencing glucose and lipid metabolism in male Wistar fatty rats and that SRCD may influence cecal fermentation. PMID- 19185779 TI - Egg yolk proteins suppress azoxymethane-induced aberrant crypt foci formation and cell proliferation in the colon of rats. AB - Dietary proteins can influence colonic carcinogenesis; some proteins have a promotional effect, whereas others exhibit a preventive effect. Dietary egg yolk proteins have been reported to suppress the expression of colon tumors in rats. In this study, we investigated the effect of consumption egg yolk proteins on cell proliferation in a rat model of azoxymethane (AOM)-induced colon cancer. We hypothesize, based on the literature of egg yolk protein actions, that they protect against colon tumor development. Therefore, male F344 rats were fed a purified AIN-93G diet containing either 20% casein (control) or 20% egg yolk proteins for 5 weeks. After 1 week on the experimental diet, the rats were administered weekly subcutaneous injections of saline or AOM for 2 weeks to induce aberrant crypt foci. Rats were administered an intraperitoneal injection of 5-bromo-2'-deoxyuridine 1 hour before being euthanized for examination of DNA synthesis in the colonic mucosa. The contents of the cecum were analyzed for the presence of short-chain fatty acids (SCFAs). In the AOM-injected rats, the yolk protein diet suppressed aberrant crypt foci formation and reduced the proliferative 5-bromo-2'-deoxyuridine-labeling index in the proximal colon when compared with the control diet. A significant increase in cecal SCFAs was observed in the rats that were fed egg yolk proteins. These results indicate that dietary egg yolk proteins have a preventive effect on AOM-induced large bowel carcinogenesis in rats; it exerts this effect by altering cell proliferation through SCFA production. This study suggests that the consumption of egg yolk proteins might be protective against colon carcinogenesis. PMID- 19185780 TI - Dietary supplementation with a combination of alpha-lipoic acid, acetyl-L carnitine, glycerophosphocoline, docosahexaenoic acid, and phosphatidylserine reduces oxidative damage to murine brain and improves cognitive performance. AB - Alzheimer disease has a complex etiology composed of nutritional and genetic risk factors and predispositions. Moreover, genetic risk factors for cognitive decline may remain latent pending age-related decline in nutrition, suggesting the potential importance of early nutritional intervention, including preventative approaches. We hypothesized that a combination of multiple nutritional additives may be able to provide neuroprotection. We demonstrate herein that dietary supplementation with a mixture of ALA, ALCAR, GPC, DHA, and PS reduced reactive oxygen species in normal mice by 57% and prevented the increase in reactive oxygen species normally observed in mice lacking murine ApoE when maintained on a vitamin-free, iron-enriched, oxidative-challenge diet. We further demonstrate that supplementation with these agents prevented the marked cognitive decline otherwise observed in normal mice maintained on this challenge diet. These findings add to the growing body of research indicating that key dietary supplementation may delay the progression of age-related cognitive decline. PMID- 19185781 TI - [Rivaroxaban: a decisive step forward in the management of postoperative thromboembolic disease?]. PMID- 19185782 TI - [Rivaroxaban: clinical pharmacology]. AB - Rivaroxaban (Xarelto) is a new oral, direct and selective inhibitor of the factor Xa of the coagulation cascade. The main pharmacokinetic characteristics of rivaroxaban are a bioavailability of approximately 80-100%, a maximum concentration obtained in 2 to 4 hours, a terminal half-life of elimination of 7 to 11 hours, a renal elimination for 1/3 for the active hepatic metabolism from the cytochrome P450 (3A4) for the other 2/3. The main sources of variability are the renal and the liver function and potential interactions with some strong inhibitors or inducers of the CYP450 3A4. Phase II clinical studies have shown that this compound can be orally administrated, once or twice daily, without any biological monitoring and without any need for dose adjustment. There is a contra indication in case of severe liver insufficiency and not recommended in case of severe renal impairment. Pharmacodynamically, Rivaroxaban is a direct and selective factor Xa inhibitor without any effect on the factor IIa and without any interaction on platelets. Four phases II with 2787 patients were carried out to for venous thromboembolic (VTE) prophylaxis after major orthopaedic surgery, showing that 10 mg once daily could be the optimal dose regimen to assess in phase III. Two phases II with 1446 patients were carried out for the treatment of VTE showing that 15 mg twice daily for 3 weeks and then 20 mg once daily could be the optimal dose regimen to evaluate in the following phases 3. No strong signal for a potential liver toxicity was shown during these 6 phases II. PMID- 19185783 TI - [Perioperative venous thromboembolism prophylaxis: short review and recommendations]. AB - The overall thromboembolic risk is the resultant of patient-related risk and surgical risk. The surgical risk is decreasing, especially with the introduction of new procedures (fast-track surgery). The value of prophylaxis has been firmly established. Mechanical prophylaxis is to be used as first-line prophylaxis when there is a risk of bleeding. Combining this with drugs increases the antithrombotic efficacy. However, the effectiveness of prophylaxis on pulmonary embolism and mortality has not been demonstrated. Renal function needs to be evaluated when low molecular weight heparins, fondaparinux, rivaroxaban or dabigatran are prescribed. An age of over 75 years and low body weight (<50 kg) have to be taken into account. There is a risk of spinal or epidural hematoma in patients receiving anticoagulants. Caution should be taken especially when administering the newer agents. Patients undergoing surgery that involves a moderate or high overall risk should receive prophylaxis until full mobilization. Patients who have undergone a total hip replacement, surgery for hip fracture, or major abdominal surgery should receive prophylaxis for about 5 weeks longer. The relevance of distal vein thromboses is debated. Surrogate venographic end-points should be gradually replaced by a combination of ultrasound and clinical criteria. The new antithrombotic agents will probably modify prevention in the years to come but currently there are very few long-term data for these products for which - it should be reminded - no antagonists are available. PMID- 19185784 TI - [Rivaroxaban (Xarelto): efficacy and safety]. AB - The oral direct Xa inhibitor rivaroxaban (Xarelto) shows great promise for prevention of venous thromboembolic events after major elective orthopedic surgery. Its consistent and predictable pharmacokinetics and pharmacodynamics across a wide range of patient populations allow administration with fixed dosing and with no coagulation monitoring. In 4 orthopaedic surgery clinical trials (12,700 patients), 10mg postoperative (6-10 hours after the end of surgery) dose, once daily, of oral rivaroxaban, achieved superior efficacy and similar safety to enoxaparin, whatever the dose of enoxaparin. Indeed, 40 mg once a day in Europe and 30 mg bid in US of enoxaparin were compared to the same dose of 10mg once daily of rivaroxaban. Furthermore, there is no difference according to liver enzymes elevation and cardio-vascular adverse events. Although the risk of spinal haematoma after neuraxial anaesthesia is rare, it is increased by concomitant use of anticoagulants. In orthopedic surgery trials with rivaroxaban to date, complications such as spinal haematoma have not been reported. The pharmacokinetic profile of rivaroxaban suggests that concurrent use with neuraxial anaesthesia should require no further precautions than currently necessary with low-molecular-weight heparin. PMID- 19185785 TI - [Summary and perspectives. Rivaroxaban]. AB - New oral anticoagulants, soon available in clinical practice, will deeply change the management of venous thromboembolism. The main advantage of these drugs is the route of administration. Moreover, among the new oral anticoagulants, rivaroxaban has a better efficacy than enoxaparin to prevent thromboembolic events after major orthopaedic surgery (THR and TKR). In phase III studies, safety profile seems adequate. A new era for prophylaxis of VTE is beginning with the new oral anticoagulants. However, improvement in the management of patients with renal failure, obese patients or elderly is needed considering that these patients have a high thrombotic and/or hemorrhagic risk. PMID- 19185787 TI - Planning and delivery of appropriate wound prevention and treatment services. PMID- 19185788 TI - The prevalence, management and outcome for acute wounds identified in a wound care survey within one English health care district. AB - This paper reports the characteristics and local management of 826 acute wounds identified during an audit across all health care providers serving the population of Bradford, UK. Of the wounds encountered 303 were traumatic wounds and 237 primary closures with smaller numbers of other acute wound types. Of the 303 traumatic wounds 174 occurred in women (57.4%). Men predominated in the under 45s (65M:26F), this being largely accounted for by hand and finger trauma (n = 62) particularly in patients of working age (M32:F12). Women predominated in the over 65s (50M:130F), this being largely accounted for by lower limb traumatic wounds (M24:F91), the majority of these being in patients 65 and over (M14:F82). In this sub-group of 96 patients 25 had wounds of 6 weeks or longer duration, only 3 had undergone Doppler assessment and only 2 received compression bandaging. Typically these wounds were of recent origin and small in size (under 1 week and less than 5 cm2 in surface area) however exceptions occurred where 10 people had wounds over 25 cm2 in area while 3 wounds had been present for over 5 years. 101 (12.2%) of the encountered wounds were considered to be infected although the practice of wound swabbing in the presence of presumed infection seemed inadequate with 37.6% of all infected acute wounds not being swabbed while 97 non-infected wounds were swabbed. Where wounds were swabbed 4.5% were found to be MRSA positive. Across all acute wound types (with the sole exception of primary closures) antimicrobial wound dressings were the most prevalent form of dressing and covered 56 (55.4%) of all infected wounds. PMID- 19185786 TI - [Rivaroxaban: mode of action]. AB - Rivaroxaban is the first oral anticoagulant with a direct anti-Xa activity to be registered (approval). As for all first comers in a class, it should be assessed both for itself and for the class. The targeting of factor-Xa factor, key component in the coagulation cascade, has the theoretical benefit of being an effective antithrombotic and a potential risk for hemorrhage, both highly dose dependent. Experience has shown us that the representativeness and predictiveness of in vitro tests and preclinical models are only partial and sometimes even misleading. This is why the responses can only come from clinical trials and rigorous research testing doses, which should be conducted specifically in all the indications foreseen, with no extrapolations. The oral anticoagulant drugs are developed in the prevention of arterial thromboembolic events caused by atrial fibrillation too, where the vitamin K antagonists (VKAs) are the current standard of care. The well-known problems of monitoring and adaptation doses with VKAs have led to developing new replacement classes without the need for control or biological adaptation. However, in certain conditions there is a need to monitor the patient. The advantage for the direct anti-Xa inhibitors such as rivaroxaban is that the prothrombin time, a routine test is sensitive and provides a prolonged response that is proportional to the plasma concentration within a wide range of concentrations. This test is potentially usable provided that the indispensable standardization is forthcoming. PMID- 19185789 TI - Oral and Maxillofacial Surgery Clinics of North America. Complications in cosmetic facial surgery. Preface. PMID- 19185790 TI - Diagnosis and management of skin resurfacing-related complications. AB - The field of skin resurfacing is undergoing rapid evolution with many new technologies that have developed, providing more choices for physicians and patients. Knowing the potential adverse effects associated with each skin resurfacing modality is paramount in selecting the appropriate approach for each candidate, thereby minimizing complications and achieving optimal results. PMID- 19185791 TI - Complications in fillers and Botox. AB - Americans spent $11 billion on cosmetic surgery procedures in 2007. Almost $3 billion were spent on minimally invasive procedures, of which injectable facial fillers and botulinum toxin type A (Botox) were the most popular. Injectables have fueled the fires of the popularity of cosmetic facial surgery. They have provided more options to patients and surgeons and provided quick, affordable, predictable, and long-lasting improvement of facial wrinkles and lip augmentation. Even in sour economies, patients want to look good and although they may not have money to spend on surgical procedures, injectable treatments remain popular. This article addresses common complications of injectable fillers and Botox. PMID- 19185792 TI - Nerve injuries and treatment in facial cosmetic surgery. AB - Surgical intervention remains a popular choice in patients seeking facial rejuvenation. Although uncommon, temporary or permanent peripheral nerve injury may complicate almost any type of invasive aesthetic procedure of the face, resulting in functional and psychological consequences for patients. Prompt recognition and appropriate intervention are necessary to avoid the long-term sequelae and improve the chances of complete neurologic recovery. Depending on the type of injury, various interventions may range from observation and close follow-up to interposition nerve grafting. This article reviews the pertinent anatomy of nerves at risk in facial cosmetic surgery and discusses various management strategies for inadvertent injury to peripheral nerves of the face. PMID- 19185793 TI - How to avoid blepharoplasty complications. AB - Complications unfortunately are part of blepharoplasty surgery. The goal of this article is to help guide the surgeon through the thought process of the preoperative evaluation, surgical planning, and actual surgery to avoid complications. With better understanding of the goals of blepharoplasty and the areas for potential problems, we hope to reduce the possibility of developing complications. PMID- 19185794 TI - Complications of neck liposuction and submentoplasty. AB - Patients desiring improved neck and jawline contours often are looking for minimally invasive procedures and are not interested in undergoing extensive face lifting procedures. Realizing the limitations, surgeons may offer their patient such procedures as liposuction and submentoplasty. Even though these procedures are less involved than a facelift, many pitfalls can occur, leading to an unfavorable result and a disappointed patient. Proper patient selection and choosing the correct operation are crucial to avoiding these situations. This article focuses on the common complications of neck liposuction and submentoplasty and reviews the management and avoidance of these complications. PMID- 19185795 TI - Complications following fat transfer. AB - Traditionally, strategies for facial rejuvenation have emphasized correction of tissue ptosis and laxity with suspensory and excisional techniques, such as face lifting and blepharoplasty. Volume loss plays a significant role in facial aging and, until recently, had not received appropriate attention. Facial fat grafting to correct volume loss has become a crucial component of facial rejuvenation in the authors' respective practices. PMID- 19185796 TI - Complications in facelift surgery and their prevention. AB - The purpose of this article is to discuss the pros and cons of various aspects of rhytidectomy surgery. The author presents his experiences that have led to performing approximately 85 facelifts a year and what has proved beneficial along the way over the past decade in his cosmetic facial practice. Because it is impossible to incorporate all aspects of facelift complications into the confines of this article, the author focuses on some of the more common problematic and avoidable complications. PMID- 19185797 TI - Complications of rhinoplasty. AB - Rhinoplasty presents a unique set of challenges for the cosmetic surgeon. Complications may arise from inadequate diagnosis, errors in surgical technique, or variations in the patient's anatomy or healing response. Complications as a result of overly aggressive surgery may also have functional consequences and be harder to correct. PMID- 19185798 TI - Complications of facial implants. AB - Facial implants are readily used for aesthetic and reconstructive efforts in lieu of autogenous materials due to obvious benefits. Alloplastic facial augmentation is not without potential complications. This article discusses the major factors that contribute to complications of facial implant surgery, based on alloplast composition, surgical technique, and facial region. Also discussed are the most common complications as well as both their prevention and management. PMID- 19185799 TI - Otoplasty complications. AB - Otoplasty, the correction of protruding ears, is a commonly performed cosmetic surgical procedure. Although few and rare, otoplasty has associated risks and complications. Most of these complications can be minimized by appropriate patient selection, careful preoperative analysis and planning, meticulous surgical technique, and compliant postoperative care. The surgeon must be familiar with the possible complications to avoid or prevent them. The astute surgeon must also be able to recognize and confidently treat adverse outcomes as they occur to minimize long-term sequelae. PMID- 19185800 TI - Complications in hair restoration surgery. AB - Hair loss affects more than 1.2 billion people worldwide. As the technology and artistry of hair restoration surgery has improved including natural results, so too has the popularity of this procedure. As with any other surgical procedure, complications may occur and this presents a major challenge for the surgeon and the patient. This article provides an overview of the complications most likely to occur during the pre, intra, and postoperative periods with modern hair transplant surgery (single follicular unit or multifollicular unit) including scalp surgery, and discusses their treatment and most importantly their prevention. PMID- 19185801 TI - How can jurors help oral and maxillofacial surgeons? AB - Oral and maxillofacial surgeons can improve patient relationships by pretending that patients are potential jurors. Looking at relationships from the eyes of a juror allows oral and maxillofacial surgeons to ascertain patients' personality types and modify communication styles accordingly. This approach also allows oral and maxillofacial surgeons to understand why patients (jurors) need to see information and evidence. PMID- 19185802 TI - A brief history of vitamin d and cancer prevention. AB - PURPOSE: To review the history of vitamin D and its use in cancer prevention. METHODS: The literature on published studies of vitamin D and its role in human health was reviewed and summarized. RESULTS: The modern history of vitamin D began in the mid-1800s, when it was noticed that city children were more likely to have rickets than rural children. Half a century later, Palm reported that children raised in sunny climates virtually never developed rickets. McCollum isolated vitamin D, and Windaus its precursors, receiving the Nobel Prize. Other scientists later observed that people with skin cancer had lower prevalence of nonskin cancers, and that lower overall mortality rates from all internal cancers combined existed in sunnier areas. These observations went largely unnoticed, and the field stagnated until 1970, when maps were created of cancer mortality rates. Through study of these maps, Cedric and Frank Garland of Johns Hopkins University reported a strong latitudinal gradient for colon cancer mortality rates in 1980, and hypothesized that higher levels of vitamin D compounds in the serum of people in the south were responsible, and that calcium intake also would reduce incidence. Edward Gorham and colleagues carried out cohort and nested studies, including the first study that found an association of a serum vitamin D compound with reduced cancer risk. William B. Grant then carried out numerous ecologic studies that extended the vitamin D-cancer theory to other cancers. CONCLUSIONS: The history of the role of vitamin D in human health is rich and much of that history is yet to be written not only by scientists, but by policy makers with the vision and leadership necessary to bridge the gap between research and policy. PMID- 19185803 TI - Reproductive factors and risk of pancreatic cancer in women: a review of the literature. AB - PURPOSE: Pancreatic cancer (PCA) is the fourth leading cause of cancer death in the United States. The male-to-female incidence and mortality ratio of PCA is 1.1 2.0. One possible explanation for this difference is that female hormone exposure is protective for the development of PCA. Several hypotheses were investigated in this systematic review: (1) increased exposure to estrogen through early menarche and later menopause is associated with a decreased risk of PCA; (2) increased exposure to pregnancy is associated with decreased risk of PCA; and (3) increased exposure to oral contraceptives and/or hormone replacement therapy is associated with decreased risk of PCA. METHODS: Of 371 articles identified, 10 case-control and 5 cohort studies met the criteria for our review. Odds ratios for case control studies and hazard ratios for cohort studies and their accompanying 95% confidence intervals for analyses relevant to our hypotheses were considered in the review. RESULTS: For all 3 hypotheses, studies displayed inconsistent results, and this may have been due to the diversity of study populations, exposure quantification, analysis approach, confounding and other limitations, and biases across studies. CONCLUSIONS: As there was no strong support for any of the 3 hypotheses, it appears that reproductive factors are not associated with the development of PCA in women. PMID- 19185805 TI - Controversies in epidemiology and policy: salt reduction and prevention of heart disease. PMID- 19185804 TI - Birth weight of offspring, maternal pre-pregnancy characteristics, and mortality of mothers: the Jerusalem perinatal study cohort. AB - PURPOSE: To explore the association between birth weight in offspring, a marker of the intrauterine environment, and mortality in their mothers, taking into account maternal pre-pregnancy characteristics, including maternal body mass index (BMI), smoking, and socioeconomic status. Distinguishing the effects of offspring's birth weight and pre-pregnancy characteristics on maternal outcome may provide clues regarding mechanisms underlying the association between birth weight and maternal mortality. METHODS: We studied long-term total mortality (average follow-up period, 29.1 years) in a population-based cohort of 13,185 mothers, aged 15 to 48 years at their offspring's birth, who delivered in West Jerusalem during 1974 through 1976. RESULTS: Univariate and multivariate Cox proportional hazard models used to estimate the hazard of overall mortality among mothers indicated a nonlinear relationship with birth weight of offspring when introduced into the models as a continuous variable, and a linear positive association with maternal pre-pregnancy BMI. Inclusion of maternal BMI and other pre-pregnancy characteristics in the model did not alter the association between offspring's birth weight and mothers' all-cause mortality. When birth weight was introduced as a categorical variable, higher mortality was observed among mothers who gave birth to babies with birth weight less than 2500 g (hazard ratio [HR] = 1.90; 95% confidence interval [95%CI], 1.23-2.94) as compared to mothers whose offspring had birth weight between 3000 and 3499 g. The HR for mothers who gave birth to babies with birth weight 4000 g or more was 1.30 (95%CI, 0.88-1.91). CONCLUSIONS: Independent of pre-pregnancy maternal BMI and other characteristics, birth weight of offspring was associated with mortality in their mothers, suggesting that intrauterine metabolic events reflected by birth weight and not explained by maternal obesity, smoking, and socioeconomic status have remote consequences for maternal health. These findings underline the need to explore specific genetic and/or environmental mechanisms that account for these associations. PMID- 19185807 TI - Hospital employee assault rates before and after enactment of the california hospital safety and security act. AB - PURPOSE: This study examines changes in violent event rates to hospital employees before and after enactment of the California Hospital Safety and Security Act in 1995. METHODS: We compared pre- and post-initiative employee assault rates in California (n = 116) emergency departments and psychiatric units with those in New Jersey (n = 50), where statewide workplace violence initiatives do not exist. Poisson regression with generalized estimating equations was used to compare assault rates between a 3-year pre-enactment period (1993-1995) and a 6-year post enactment period (1996-2001) using New Jersey hospitals as a temporal control. RESULTS: Assault rates among emergency department employees decreased 48% in California post-enactment, compared with emergency department employee assault rates in New Jersey (rate ratio [RR] = 0.52, 95% confidence interval [CI]: 0.31, 0.90). Emergency department employee assault rates decreased in smaller facilities (RR = 0.46, 95% CI: 0.21, 0.96) and for-profit-controlled hospitals (RR = 0.39, 95% CI: 0.19, 0.79) post-enactment. Among psychiatric units, for profit-controlled hospitals (RR = 0.41, 95% CI: 0.19, 0.85) and hospitals located in smaller communities (RR = 0.44, 95% CI: 0.21, 0.92) experienced decreased assault rates post-enactment. CONCLUSION: Policy may be an effective method to increase safety to health care workers. PMID- 19185806 TI - Breast cancer survival in ontario and california, 1998-2006: socioeconomic inequity remains much greater in the United States. AB - This study re-examined the differential effect of socioeconomic status on the survival of women with breast cancer in Canada and the United States. Ontario and California cancer registries provided 1,913 cases from urban and rural places. Stage-adjusted cohorts (1998-2000) were followed until 2006. Socioeconomic data were taken from population censuses. SES-survival associations were observed in California, but not in Ontario, and Canadian survival advantages in low-income areas were replicated. A better controlled and updated comparison reaffirmed the equity advantage of Canadian health care. PMID- 19185808 TI - Overall obesity and abdominal adiposity as predictors of mortality in u.s. White and black adults. AB - PURPOSE: The association of overall obesity and abdominal adiposity in predicting risk of all-cause mortality in white and black adults was compared. METHODS: This prospective study included a national sample of 3219 non-Hispanic white and 2,561 non-Hispanic black adults 30 to 64 years of age enrolled in the third National Health and Nutrition Examination Survey during 1988-1994. Multiple measures of adiposity were measured and calculated, including body mass index (BMI), waist circumference, waist-to-hip ratio (WHR), and waist-to-thigh ratio (WTR). Vital status was ascertained with the National Death Index through 2000. RESULTS: During 12 years of follow-up (51,133 person-years), 188 white and 222 black adults died. After adjustment for age, education, smoking, and existing disease, positive dose-response associations between WTR and mortality in both sexes and races, and WHR in white and black women were observed (p(trend) < 0.05 for all). These results were unchanged after additional adjustment for BMI. In contrast, BMI and waist circumference alone exhibited curvilinear-shaped associations with mortality. A higher WTR was associated with a higher risk of mortality among normal weight (BMI: 18.5-24.9 kg/m(2)) and obese (BMI: > or =30.0 kg/m(2)) white and black adults. CONCLUSIONS: These results suggest ratio measures of abdominal adiposity, particularly WTR in both sexes and WHR in women, strongly and positively predict mortality, independent of BMI, among white and black adults. Furthermore, WTR offers additional prognostic information beyond that provided by BMI alone. PMID- 19185809 TI - Implications of myocardial matrix remodeling by adipokines in obesity-related heart failure. AB - Owing to the increased incidence of obesity and its association with heart failure, there is now great interest in elucidating the underlying molecular mechanisms linking these pathologies. Since the discovery of adipose-derived hormones and cytokines, their important regulatory role in myocardial function has emerged. The events that these adipokines can regulate include alterations in myocardial metabolism, cardiomyocyte hypertrophy, cell death, and structure and composition of the extracellular matrix. Here, we focus on the last of these and review current research demonstrating an important role for adipokines, with particular emphasis on leptin and adiponectin, in regulating matrix metalloproteinases, tissue inhibitors of matrix metalloproteinases, and collagens. From this, it is clear that adipokines are capable of contributing to remodeling of the myocardial extracellular matrix, and the altered adipokine profiles observed in obese individuals may be important in the pathogenesis of heart failure. The feasibility of adipokine manipulation as a potential therapeutic treatment in preventing maladaptive cardiac remodeling is also discussed. PMID- 19185810 TI - Embryonic endothelial progenitor cell-mediated cardioprotection requires Thymosin beta4. AB - Myocardial damage is frequently occurring upon a prolonged period of ischemia, although subsequent reperfusion as standard therapy is established. Among the pleiotropic causes of ischema-reperfusion injury, loss of cardiomyocytes, microcirculatory disturbances, and postischemic inflammation have been frequently observed. Current clinical cell therapy after acute myocardial mostly aims at neovascularization and enhancement of tissue repair, whereas acute cardioprotection after ischemia and reperfusion has rarely been studied. Recently, embryonic endothelial progenitor cells (eEPCs) have been found to provide cardioprotection against acute ischemia-reperfusion injury (24 hours) in a preclinical pig model. The paracrine effect of eEPCs was mimicked by regional application of a single, highly expressed protein, Thymosin beta4. This review focuses on underlying mechanisms of acute cardioprotection provided by eEPCs and, in particular, Thymosin beta4. PMID- 19185811 TI - Use of rapid prototyping in the care of patients with structural heart disease. AB - Advances in surgery, interventional techniques, and critical care have allowed more than 90% of children born with structural heart defects to survive into adulthood. In addition, advances in imaging technology continue to raise awareness of hemodynamically significant intracardiac shunt lesions in both adults and children. Adult cardiologists are now faced with the daunting task of caring for patients with complex structural heart lesions, a population subset that at one time was exclusively cared for by pediatric cardiologists and congenital heart disease specialists. Given the wide range of anatomic complexity present in patients with structural heart disease, the definition and anatomic clarification of their structural abnormalities through high-quality noninvasive imaging has become paramount. Current two-dimensional imaging techniques, however, remain limited in their ability to effectively illustrate the complex three-dimensional relationships present in structural heart disease. Rapid prototyping, a process by which three-dimensional digital surface models are converted into physical models, represents the next evolution in advanced image processing and may serve as a means to improve our understanding of the many forms of structural heart disease. Ultimately, the technology may be used to enhance the level of care provided to the growing number of patients with structural heart defects. We recently reviewed the novel cardiovascular application of rapid prototyping. This review examines the expanded applications of rapid prototyping in the care and treatment of adult patients with structural heart disease. PMID- 19185812 TI - Congenital long-QT syndromes: a clinical and genetic update from infancy through adulthood. AB - Long-QT syndromes (LQTSs) have been described in all ages and are a significant cause of cardiovascular mortality, especially in structurally normal hearts. Abnormalities in transmembrane ion conduction channels and structural proteins produce these clinical syndromes, labeled LQT1-LQT12; however, genotype-positive patients still represent only about 70% of LQTSs. Future research will determine the etiology of the remaining cases, further risk-stratify the known genetic defects, improve current treatment options for these syndromes, and uncover novel therapies. PMID- 19185813 TI - Foxc2 transcription factor: a newly described regulator of angiogenesis. AB - Angiogenesis is a critical process to form new blood vessels from preexisting vessels under physiologic and pathologic conditions and involves cellular and morphologic changes such as endothelial cell proliferation, migration, and vascular tube formation. Despite evidence that angiogenic factors, including vascular endothelial growth factor and Notch, control various aspects of angiogenesis, the molecular mechanisms underlying gene regulation in blood vessels and surrounding tissues are not fully understood. Importantly, recent studies demonstrate that Forkhead transcription factor Foxc2 directly regulates expression of various genes involved in angiogenesis, CXCR4, integrin beta3, Delta-like 4 (Dll4), and angiopoietin 2, thereby controlling angiogenic processes. Thus, Foxc2 is now recognized as a novel regulator of vascular formation and remodeling. This review summarizes current knowledge about the function of Foxc2 in angiogenesis and discusses prospects for future research in Foxc2-mediated pathologic angiogenesis in cardiovascular disease. PMID- 19185815 TI - National Priorities Partnership: addressing challenges in the health care system. PMID- 19185816 TI - ASPAN's Delphi study on national research: priorities for perianesthesia nurses in the United States. AB - Prudent nursing practice mandates the best, scientific evidence available. The complexity of perianesthesia practice, management, and education, coupled with required competencies within these domains, generate problems demanding answers. ASPAN's financial and human resources, however, are limited. Therefore, ASPAN must be able to articulate research priorities. The purpose of this study was to identify and prioritize the research questions related to perianesthesia practice, management, and education. A three-round Delphi study conducted with perianesthesia nursing experts determined research priorities for perianesthesia nursing practice. Research problems were identified and refined into research questions. Questions were ranked for their priority. Research priorities focused on the following themes: evidence supporting ASPAN standards, staffing ratios, nurse fatigue, adverse patient outcomes, patient care issues related to intensive care unit (ICU) overflow, obstructive sleep apnea, pain management, glycemic control, and clinical critical care competencies. This study advances the mission of ASPAN by identifying the top national perianesthesia research priorities related to practice, management, and education, and should guide researchers in their continued exploration of the science of perianesthesia nursing. PMID- 19185814 TI - Monocyte-endothelial cell interactions in the development of atherosclerosis. AB - The activation of endothelial cells at atherosclerotic lesion-prone sites in the arterial tree results in the up-regulation of cell adhesion molecules and chemokines, which mediate the recruitment of circulating monocytes. Accumulation of monocytes and monocyte-derived phagocytes in the wall of large arteries leads to chronic inflammation and the development and progression of atherosclerosis. This review discusses the nature of these molecules and the mechanisms involved in the early steps of monocyte recruitment into atherosclerotic lesion sites within the vessel wall. PMID- 19185817 TI - Does audiovisual stimulation with music and nature sights (MuViCure) reduce pain and discomfort during placement of a femoral nerve block? AB - MuViCure (Photobia ApS, Copenhagen, Denmark) is a new program for audiovisual stimulation. We hypothesized that audiovisual stimulation would reduce pain and discomfort and improve patients' well-being during placement of a femoral nerve block. Fifty-five outpatients scheduled for anterior cruciate ligament reconstruction were randomly allocated into three groups: the first group received audiovisual stimulation (MuViCure), the second group received audio stimulation (MusiCure, Gefion Records ApS, Virum, Denmark), and the third group received no intervention (control). Ten of the 55 patients underwent a qualitative in-depth interview 1-2 days after surgery. Pain and discomfort during the procedure were more prominent in the MuViCure group when compared with the other two groups. Despite these negative results, 14 of the 19 patients in the MuViCure group answered that MuViCure had a positive effect on their well-being. The qualitative interviews revealed that a number of factors other than the audiovisual stimulation had a significant impact on the patients' experience. The use of MuViCure may be more appropriate in other settings. PMID- 19185818 TI - Scientific advances in the genetic understanding and diagnosis of malignant hyperthermia. AB - Malignant hyperthermia (MH), a potentially fatal disorder triggered by certain types of general anesthesia, has received much attention in the scientific literature. From the first case report in 1960 until the present, hundreds of studies have been conducted. The diagnosis of MH has evolved from subjective assumptions by family history and clinical diagnosis to more sophisticated laboratory testing. A genetic basis for MH was recognized in the early 1990s and, since then, complex genetic pathways have been demonstrated. The purpose of this paper is to summarize the research literature on what is known scientifically about the diagnosis and genetic basis of MH. PMID- 19185819 TI - Transfusion-related acute lung injury. AB - Approximately one person in 5,000 will experience an episode of transfusion related acute lung injury (TRALI) in conjunction with the transfusion of whole blood or blood components. Its hallmarks include hypoxemia, dyspnea, fever, hypotension, and bilateral pulmonary edema (noncardiogenic). The mortality for reported cases is 16.3%. The incidence and mortality may be even higher than estimated because of under-recognition and under-reporting. Although TRALI was identified as a clinical entity in the 1980s, a lack of consensus regarding a definition was present until 2004. An exact cause has yet to be identified; however, there are two theories regarding the etiology: the "antibody" and the "two-hit" theories. These theories involve both donor and recipient factors. Further education and research are needed to assist in the development of strategies for the prevention and treatment of TRALI. PMID- 19185820 TI - A systematic approach to chest x-ray interpretation in the perianesthesia unit. AB - In the perianesthesia setting, the chest X-ray has become an essential diagnostic tool to help direct the optimal care of critically ill patients. The ordering of medical tests, the interpretation of the results, and the determination of an appropriate medical plan of care remain within the role of the physician. In many United States acute care hospitals, however, the perianesthesia nurse is taught to implement emergency standing orders including obtaining a chest X-ray for unexpected respiratory distress, unexplained hypoxemia, or unequal breath sounds. This article will offer the perianesthesia nurse the basic knowledge to rapidly and systematically read chest radiographs and compare the findings with the patient's previous films and current clinical presentation. The assessment tool outlined in this paper may be combined with the clinical picture the patient presents to ensure that the nurse implements needed interventions in a timely manner. PMID- 19185821 TI - Challenges in pain assessment. PMID- 19185822 TI - Considering the human factors in patient safety. PMID- 19185824 TI - A new year of hope. PMID- 19185825 TI - Protecting the health of a nation: spotting the sentinel signs of disease. PMID- 19185826 TI - Multidisciplinary team meetings: what does the future hold for the flies raised in Wittgenstein's bottle? PMID- 19185827 TI - Palonosetron: more than just another option? PMID- 19185828 TI - A new subgroup of high-risk acute lymphoblastic leukaemia. PMID- 19185829 TI - Tibolone: the risk is too high. PMID- 19185830 TI - Early T-cell precursor acute lymphoblastic leukaemia. PMID- 19185831 TI - It's not palliative care, it's palliative treatment. PMID- 19185833 TI - Emergency surgery in the era of molecular treatment of solid tumours. AB - In the advancing era of molecular therapy of solid tumours, emergency treatment of complications, such as bowel perforation, haemorrhage, and tumour rupture, is likely to evolve into one of the main challenges of surgical oncology. These complications might be caused by disease progression from resistance to therapy, side-effects of therapy on normal vasculature, and therapeutic induction of excessively responding tumours. This Review outlines the probability and management of emergency operations during molecularly targeted therapy of solid tumours. Special attention is given to advanced gastrointestinal stromal tumours and colorectal cancer, and therapy with imatinib, sunitinib, and bevacizumab. PMID- 19185834 TI - Palliating delirium in patients with cancer. AB - Delirium is a frequent complication in oncology. Its definition as a disorder of consciousness, attention, and cognition is useful to elaborate a rational framework of its pathophysiology and to interpret the role of different aetiological factors and therapeutic interventions. Many aetiologies and an interaction between risk and predisposing factors have been shown to contribute to most cases of delirium. A screening of potential aetiologies is always mandatory to benefit reversible cases. The palliative treatment of symptoms of delirium includes non-pharmacological, environmental, and preventive interventions and the use of haloperidol. If haloperidol fails to control delirium, sedation with other drugs can be necessary. Specific attention to the qualitative aspects of care and to the effect of delirium on family members should be given in the overall assessment of the patient in his or her cancer trajectory. PMID- 19185835 TI - Alcohol and genetic polymorphisms: effect on risk of alcohol-related cancer. AB - Public health guidelines aim to limit the consumption of alcoholic beverages worldwide and the subsequent health burden. In particular, alcohol consumption is an avoidable risk factor for cancer. In human beings, ethanol in alcoholic drinks is mainly oxidised in the liver by alcohol dehydrogenases to acetaldehyde, and is further detoxified to acetate by aldehyde dehydrogenases. Functional variants in genes involved in alcohol metabolism result in differences between individuals in exposure to carcinogenic acetaldehyde, suggesting a possible interaction of genetic susceptibility and alcohol exposure in cancer. We reviewed available studies of the combined effects of alcohol drinking and genetic polymorphisms on alcohol-related cancer risk. Most available data were for polymorphisms in alcohol and folate metabolism. We give an overview of published studies on the combined effects of alcohol drinking and polymorphisms in genes for alcohol dehydrogenase (ADH), aldehyde dehydrogenase (ALDH), cytochrome P450 2E1, and methylene-tetrahydrofolate reductase on the risk of alcohol-related cancer. Current data lend support to a role of polymorphisms ADH1B and ALDH2 combined with alcohol consumption in cancer. Other available data are insufficient or inconclusive, highlighting the need for additional studies. PMID- 19185836 TI - Germ-line DNA polymorphisms and susceptibility to differentiated thyroid cancer. AB - The incidence of differentiated thyroid carcinoma (DTC) is increasing in most developed countries. The only well-known risk factor for thyroid cancer is exposure to ionising radiation. DTC is characterised by a strong hereditability, and individual susceptibility is likely due to genetic factors modulating the environmental risk. Identification of genetic polymorphisms is important for understanding the potential mechanisms involved in thyroid carcinogenesis. In this Review, we assess epidemiological data on the role of germ-line DNA polymorphisms and the risk of DTC. We have included only case-control studies that compare the incidence of germ-line mutations in patients with DTC, with the incidence of mutations in patients without a history of DTC. Such an analysis of genetic susceptibility in differentiated thyroid cancer has not yet been published. PMID- 19185837 TI - Preoperative treatment and surgery in gastric cancer: friends or foes? AB - Until 2006, most reports of preoperative (neoadjuvant) treatments for gastric carcinoma were inconclusive and produced confusing results due to inhomogeneous treatment regimens, selection of patients, and response assessment. Since publication of the results from the MAGIC trial, substantial scientific evidence has suggested the benefits of perioperative (preoperative and postoperative) chemotherapy to locally advanced gastric cancer. To date, this phase III trial, coupled with preliminary data from other published reports on neoadjuvant chemotherapy and radiotherapy, supports the theoretical advantages of preoperative treatment for gastric carcinoma, thus introducing the concept of delayed surgery. Neoadjuvant treatment of resectable, locally advanced tumours might improve patients' outcomes and postpone the need for curative resection, but it also exposes patients to the risk of tumour progression. PMID- 19185838 TI - Protons for radiotherapy: a 1946 proposal. PMID- 19185839 TI - Inflammation and cancer: IL-6 and STAT3 complete the link. AB - There is growing evidence that tumors are sustained and promoted by inflammatory signals from the surrounding microenvironment. Two papers by Grivennikov et al. and Bollrath et al. in this issue of Cancer Cell demonstrate the importance of the interleukin-6 family of proinflammatory cytokines and their downstream effector STAT3 in colitis-associated colon cancer. PMID- 19185840 TI - Reinforcing suppression using regulators: a new link between STAT3, IL-23, and Tregs in tumor immunosuppression. AB - STAT3 plays many roles in tumorigenesis. In this issue of Cancer Cell, Kortylewski et al. show that in the tumor microenvironment, STAT3 enhances the expression of the protumor cytokine IL-23 in macrophages but inhibits the antitumor cytokine IL-12 in dendritic cells. STAT3 also mediates IL-23's effect of activating tumor-infiltrating regulatory T cells. PMID- 19185841 TI - Identifying the perpetrator in medulloblastoma: Dorian Gray versus Benjamin Button. AB - Tumors contain a subpopulation of tumor-propagating cells (TPCs) that are critical for their growth. In this issue, Read, Wechsler-Reya, and colleagues show that in an animal model of medulloblastoma, TPCs express the surface marker CD15 and have properties distinct from neural stem cells. PMID- 19185842 TI - T cell acute lymphoblastic leukemia: NOTCHing the way toward a better treatment outcome. AB - Gamma-secretase inhibitors block the activation of NOTCH1 but have limited activity against T cell acute lymphoblastic leukemia (T-ALL) and cause severe gastrointestinal toxicity. In a recent study, Real et al. show that a potent gamma-secretase inhibitor potentiates the cytotoxicity of dexamethasone against glucocorticoid-resistant T-ALL cells, while dexamethasone abrogates the gastrointestinal toxicity induced by the gamma-secretase inhibitor. PMID- 19185843 TI - The stem of cancer. AB - Despite great advances in our understanding of tumor initiation and progression, the identity of the "cell of origin" of cancer remains elusive. Two recent publications provide experimental evidence that normal intestinal stem cells are the cells of origin of intestinal cancer in the mouse. PMID- 19185844 TI - gp130-mediated Stat3 activation in enterocytes regulates cell survival and cell cycle progression during colitis-associated tumorigenesis. AB - Although gastrointestinal cancers are frequently associated with chronic inflammation, the underlying molecular links have not been comprehensively deciphered. Using loss- and gain-of-function mice in a colitis-associated cancer model, we establish here a link comprising the gp130/Stat3 transcription factor signaling axis. Mutagen-induced tumor growth and multiplicity are reduced following intestinal epithelial cell (IEC)-specific Stat3 ablation, while its hyperactivation promotes tumor incidence and growth. Conversely, IEC-specific Stat3 deficiency enhances susceptibility to chemically induced epithelial damage and subsequent mucosal inflammation, while excessive Stat3 activation confers resistance to colitis. Stat3 has the capacity to mediate IL-6- and IL-11 dependent IEC survival and to promote proliferation through G1 and G2/M cell cycle progression as the common tumor cell-autonomous mechanism that bridges chronic inflammation to tumor promotion. PMID- 19185845 TI - IL-6 and Stat3 are required for survival of intestinal epithelial cells and development of colitis-associated cancer. AB - Colitis-associated cancer (CAC) is the most serious complication of inflammatory bowel disease. Proinflammatory cytokines have been suggested to regulate preneoplastic growth during CAC tumorigenesis. Interleukin 6 (IL-6) is a multifunctional NF-kappaB-regulated cytokine that acts on epithelial and immune cells. Using genetic tools, we now demonstrate that IL-6 is a critical tumor promoter during early CAC tumorigenesis. In addition to enhancing proliferation of tumor-initiating cells, IL-6 produced by lamina propria myeloid cells protects normal and premalignant intestinal epithelial cells (IECs) from apoptosis. The proliferative and survival effects of IL-6 are largely mediated by the transcription factor Stat3, whose IEC-specific ablation has profound impact on CAC tumorigenesis. Thus, the NF-kappaB-IL-6-Stat3 cascade is an important regulator of the proliferation and survival of tumor-initiating IECs. PMID- 19185846 TI - Regulation of the IL-23 and IL-12 balance by Stat3 signaling in the tumor microenvironment. AB - Interactions between tumor and immune cells either enhance or inhibit cancer progression. We show here that Stat3 signaling within the tumor microenvironment induces a procarcinogenic cytokine, IL-23, while inhibiting a central anticarcinogenic cytokine, IL-12, thereby shifting the balance of tumor immunity toward carcinogenesis. Stat3 induces expression of IL-23, which is mainly produced by tumor-associated macrophages, via direct transcriptional activation of the IL-23/p19 gene. Furthermore, Stat3 inhibits NF-kappaB/c-Rel-dependent IL 12/p35 gene expression in tumor-associated dendritic cells. Tumor-associated regulatory T cells (Tregs) express IL-23 receptor, which activates Stat3 in this cell type, leading to upregulation of the Treg-specific transcription factor Foxp3 and the immunosuppressive cytokine IL-10. These results demonstrate that Stat3 promotes IL-23-mediated procarcinogenic immune responses while inhibiting IL-12-dependent antitumor immunity. PMID- 19185847 TI - Orai1 and STIM1 are critical for breast tumor cell migration and metastasis. AB - Tumor metastasis is the primary cause of death of cancer patients. Understanding the molecular mechanisms underlying tumor metastasis will provide potential drug targets. We report here that Orai1 and STIM1, both of which are involved in store operated calcium entry, are essential for breast tumor cell migration in vitro and tumor metastasis in mice. Reduction of Orai1 or STIM1 by RNA interference in highly metastatic human breast cancer cells or treatment with a pharmacological inhibitor of store-operated calcium channels decreased tumor metastasis in animal models. Our data demonstrate a role for Orai1 and STIM1 in tumor metastasis and suggest store-operated calcium entry channels as potential cancer therapeutic targets. PMID- 19185848 TI - Identification of CD15 as a marker for tumor-propagating cells in a mouse model of medulloblastoma. AB - The growth of many cancers depends on self-renewing cells called cancer stem cells or tumor-propagating cells (TPCs). In human brain tumors, cells expressing the stem cell marker CD133 have been implicated as TPCs. Here we show that tumors from a model of medulloblastoma, the Patched mutant mouse, are propagated not by CD133(+) cells but by cells expressing the progenitor markers Math1 and CD15/SSEA 1. These cells have a distinct expression profile that suggests increased proliferative capacity and decreased tendency to undergo apoptosis and differentiation. CD15 is also found in a subset of human medulloblastomas, and tumors expressing genes similar to those found in murine CD15(+) cells have a poorer prognosis. Thus, CD15 may represent an important marker for TPCs in medulloblastoma. PMID- 19185850 TI - Redistribution of Rab27-specific effector Slac2-c, but not Slp4-a, after isoproterenol-stimulation in rat parotid acinar cells. AB - Small GTPase Rab27 has been implicated in the regulation of different types of membrane trafficking, including melanosome transport and various regulated secretion events. We have previously shown that Rab27 and its effectors, Slac2 c/MyRIP and Slp4-a/granuphilin-a, are involved in the control of isoproterenol (IPR)-induced amylase release from rat parotid acinar cells. The ability of Rab to interact with the specific effectors is important. However, little is known about the fate of these effectors after beta-adrenergic stimulation in parotid acinar cells. The present study investigated changes in intracellular redistribution of Slac2-c and Slp4-a in parotid acinar cells after IPR treatment. Subcellular fractionation studies detected Slac2-c and Slp4-a in the apical plasma membrane (APM) and secretory granules under resting conditions. After 5min of IPR treatment, Slac2-c was rapidly recruited to the luminal site, but after 30 min, the amount of Slac2-c in the APM fraction was reduced by approximately 80% compared to the increased level after 5 min of IPR treatment. Such reductions in Slac2-c are likely caused by the translocation of Slac2-c from the APM to the cytosol. In addition, we found that Slac2-c in the cytosolic fraction, but not other fractions, disappeared in the presence of Ca(2+). Since Slac2-c contains multiple PEST-like sequences (i.e., potential signals for rapid protein degradation), we suggest that Slac2-c is Ca(2+)-dependently proteolyzed in the cytosol after exocytosis. In contrast, intracellular localization and expression levels of Slp4-a in parotid acinar cells were unaltered even after beta stimulation, indicating completely different fates for the two Rab27 effectors after beta-stimulation. PMID- 19185849 TI - mTOR complex 2 is required for the development of prostate cancer induced by Pten loss in mice. AB - mTOR complex 2 (mTORC2) contains the mammalian target of rapamycin (mTOR) kinase and the Rictor regulatory protein and phosphorylates Akt. Whether this function of mTORC2 is critical for cancer progression is unknown. Here, we show that transformed human prostate epithelial cells lacking PTEN require mTORC2 to form tumors when injected into nude mice. Furthermore, we find that Rictor is a haploinsufficient gene and that deleting one copy protects Pten heterozygous mice from prostate cancer. Finally, we show that the development of prostate cancer caused by Pten deletion specifically in prostate epithelium requires mTORC2, but that for normal prostate epithelial cells, mTORC2 activity is nonessential. The selective requirement for mTORC2 in tumor development suggests that mTORC2 inhibitors may be of substantial clinical utility. PMID- 19185851 TI - Small salivary gland size in patients with xerostomia of unknown etiology. AB - OBJECTIVE: We have recently reported that flow rates of whole saliva in young healthy humans correlate positively with salivary gland sizes. The low rate of salivary secretion in xerostomia patients may be related to the small size of the salivary glands. To investigate this possibility, relationships between salivary secretions and salivary gland sizes were investigated in unknown-etiology xerostomia patients and healthy controls. DESIGN: The sizes of the three major salivary glands in seven xerostomia patients and seven age- and gender-matched healthy controls who have no previous disease history and prescription medication related to xerostomia, were measured by use of a magnetic resonance imaging technique. The salivary glands of all subjects failed to show any pathological aspects in magnetic resonance images. The flow rates of unstimulated and chewing stimulated whole saliva were also measured. RESULTS: Flow rates of unstimulated and chewing-stimulated whole saliva and the sizes of the parotid and submandibular glands were significantly lower and smaller in xerostomia patients of unknown etiology when compared with healthy controls. In addition, salivary flow rates per size of the combined three major salivary glands were also significantly lower in the xerostomia patients of unknown etiology. CONCLUSIONS: The smaller salivary gland size in xerostomia patients of unknown etiology may be one of the causes of the reduced salivary secretion. The secretion rates as a function of gland sizes were also lower, and so it is likely that functional impairments of the salivary gland are also present in patients with xerostomia of unknown etiology. PMID- 19185852 TI - Kinetically controlled Ferrier rearrangement of 3-O-mesyl-D-glycal derivatives. AB - The Ferrier rearrangement, which is widely used in carbohydrate chemistry, is generally performed under acidic conditions to give an alpha anomer with high stereoselectivity. We have found that 3-O-mesyl-D-glycals 2-4 were smoothly reacted with alcohols in the presence of triethylamine. The present reaction was shown to proceed under kinetic control to give approximately 1.3:1.0 mixture of alpha and beta anomers, indicating that a kinetic anomeric effect does not operate. PMID- 19185853 TI - Structure of the O-polysaccharide from the lipopolysaccharide of Providencia alcalifaciens O31 containing an ether of D-mannose with (2R,4R)-2,4 dihydroxypentanoic acid. AB - Recently, ether-linked diastereomeric 2,4-dihydroxypentanoic acids have been reported as new components of bacterial glycans [Shashkov, A. S. et al.Nat. Prod. Commun.2008, 3, 1625-1630]. In this work, an ether of (2R,4R)-2,4 dihydroxypentanoic acid (Dhpa) with d-mannose was identified in the O polysaccharide of Providencia alcalifaciens O31, and the polysaccharide structure was elucidated. Studies by NMR spectroscopy confirmed the ether linkage between O 2 of Dhpa and O-4 of Man, and the absolute configuration of Man was determined after ether cleavage with boron trichloride. In the polysaccharide, Dhpa was found to exist partially in the form of 1,4-lactone. Using sugar and methylation analyses along with (1)H and (13)C NMR spectroscopy, including 2D (1)H,(1)H COSY, TOCSY, ROESY, H-detected (1)H,(13)C HSQC, and gHMBC experiments, the following structure of the tetrasaccharide repeating unit of the polysaccharide was established: [carbohydrate structure: see text] PMID- 19185854 TI - Sources of information for discriminating dynamic human actions. AB - Despite the importance of action identification and discrimination in action perception and social cognition more broadly, little research has investigated how these processes are achieved. To this end, we sought to identify the extent to which adults capitalize on featural versus configural sources of information when discriminating small-scale human actions such as grasp and place. Results across two experiments indicate adults are sensitive to both sources of information in action discrimination, but selectively attend to featural over configural action information. The findings also parallel what is known regarding face processing: processing of configural information is especially disrupted by inversion, whereas processing of featural information is specifically affected by low-pass filtering. PMID- 19185855 TI - Ensembled support vector machines for human papillomavirus risk type prediction from protein secondary structures. AB - Infection by the human papillomavirus (HPV) is regarded as the major risk factor in the development of cervical cancer. Detection of high-risk HPV is important for understanding its oncogenic mechanisms and for developing novel clinical tools for its diagnosis, treatment, and prevention. Several methods are available to predict the risk types for HPV protein sequences. Nevertheless, no tools can achieve a universally good performance for all domains, including HPV and nor do they provide confidence levels for their decisions. Here, we describe ensembled support vector machines (SVMs) to classify HPV risk types, which assign given proteins into high-, possibly high-, or low-risk type based on their confidence level. Our approach uses protein secondary structures to obtain the differential contribution of subsequences for the risk type, and SVM classifiers are combined with a simple but efficient string kernel to handle HPV protein sequences. In the experiments, we compare our approach with previous methods in accuracy and F1 score, and present the predictions for unknown HPV types, which provides promising results. PMID- 19185856 TI - Virtual reality prototype for measurement of expression characteristics in emotional situations. AB - Expressions are a basic necessity for daily living, as they are required for managing relationships with other people. Conventional expression training has difficulty achieving an objective measurement, because their assessment depends on the therapist's ability to assess a patient's state or training effectiveness. In addition, it is difficult to provide emotional and social situations in the same manner for each training or assessment session. Virtual reality techniques can overcome shortcomings occurring in conventional studies by providing exact and objective measurements and emotional and social situations. In this study, we developed a virtual reality prototype that could present emotional situation and measure expression characteristics. Although this is a preliminary study, it could be considered that this study shows the potential of virtual reality as an assessment tool. PMID- 19185857 TI - Effect of high environmental temperature on semen parameters among fertile men. AB - OBJECTIVE: To evaluate the effect of high environmental occupational temperature on semen parameters of fertile men. DESIGN: Prospective. SETTING: Steel-casting plant. PATIENT(S): Ninety fertile workers exposed to a high temperature compared with 40 fertile workers working under ordinary conditions as control subjects. INTERVENTION(S): Measurement of scrotal temperature by invagination thermometry, air temperature, relative humidity by aspirated psychrometer, radiant heat by globe thermometer, air velocity by light vane anemometer, and semen analysis. MAIN OUTCOME MEASURE(S): Scrotal temperature and semen analysis. RESULT(S): Nonsignificant difference was found between the two groups regarding their scrotal temperature. Also, nonsignificant differences were demonstrated regarding semen analysis parameters being in the normozoospermic range. CONCLUSION(S): Under high environmental temperature, semen parameters were within normozoospermic levels owing to body acclimatization mechanisms. PMID- 19185858 TI - A 1-year longitudinal study of psychological morbidity after miscarriage. AB - OBJECTIVE: To examine the 1-year longitudinal course of psychological outcomes after miscarriage. DESIGN: Longitudinal observational study. SETTING: University affiliated teaching hospital. PATIENT(S): 280 miscarrying women and 150 nonpregnant women. INTERVENTION(S): Psychological outcomes were assessed using the 12-item General Health Questionnaire (GHQ-12) and Beck Depression Inventory (BDI) immediately, 3 months, 6 months, and 12 months after miscarriage. MAIN OUTCOME MEASURE(S): Scores on GHQ-12 and BDI. RESULT(S): Half (55%) of the miscarrying women scored high (>or=4) on the GHQ-12 immediately, 25% at 3 months; 17.8% at 6 months, and 10.8% at 1 year after miscarriage; 26.8% of the patients scored high on the BDI (>or=12) immediately, 18.4% at 3 months, 16.4% at 6 months, and 9.3% at 1 year after miscarriage. Patients who were initially more distressed continued to score higher on both the GHQ-12 and the BDI along the 1 year course when compared with those who were initially less distressed. When compared with the nonpregnant controls, the miscarrying women scored statistically significantly higher on the GHQ-12 and BDI; the differences became not statistically significant only 1 year after miscarriage. CONCLUSION(S): A statistically significant proportion of patients reported psychological morbidity shortly after miscarriage, but their level of distress reduced over time until they were comparable with the controls 1 year later. Patients who were initially more distressed continued to be throughout the 1-year course. PMID- 19185859 TI - True osseous metaplasia of the endometrium: the bone is not from a fetus. AB - OBJECTIVE: To identify the origin of calcified tissue in endometrial ossification. DESIGN: DNA analyses from the ossified tissue and from the woman were studied to compare both genotypes. SETTING: University and general hospitals. PATIENT(S): A 27-year-old infertile woman diagnosed of osseous metaplasia of the endometrium. INTERVENTION(S): Hysteroscopic resection of the endometrial osseous metaplasia for DNA analysis. MAIN OUTCOME MEASURE(S): DNA comparison between the patient and the osseous tissue extracted from the uterus. RESULT(S): All markers produced the same allele length for both blood and endometrial biopsy (including bones), thus confirming the same genetic origin. CONCLUSION(S): Endometrial ossification is derived from the patient, resulting in a true osseous metaplasia. PMID- 19185860 TI - Association of adolescent obesity and lifetime nulliparity--the Study of Women's Health Across the Nation (SWAN). AB - OBJECTIVE: To evaluate whether adolescent obesity is associated with difficulties in becoming pregnant later in life. DESIGN: Cross-sectional analysis of baseline data from a longitudinal cohort. SETTING: Multiethnic, community-based observational study of U.S. women. PATIENT(S): Three thousand one hundred fifty four midlife women. MAIN OUTCOME MEASURE(S): Lifetime nulliparity and lifetime nulligravidity. RESULT(S): Five hundred twenty-seven women (16.7%) women had never delivered a baby. Participants were categorized by self-reported high school body mass index (BMI): underweight (<18.5 kg/m(2)), normal (18.5-24.9 kg/m(2)), overweight (25-29.9 kg/m(2)), and obese (>30 kg/m(2)). The prevalence of lifetime nulliparity increased progressively across the high school BMI categories: 12.7%, 16.7%, 19.2%, and 30.9%, respectively. Multivariable logistic regression analysis confirmed that women who were obese adolescents had significantly higher odds of remaining childless compared with normal weight women (odds ratio [OR] 2.84; 95% confidence interval [CI], 1.59-5.10) after adjusting for adult BMI, history of nongestational amenorrhea, marital status, ethnicity, study site, and measures of socioeconomic status. Furthermore, adolescent obesity was associated with lifetime nulligravidity (OR = 3.93; 95% CI, 2.12-7.26). CONCLUSION(S): Adolescent obesity is associated with lifetime nulliparity and nulligravidity in midlife U.S. women. PMID- 19185861 TI - Estrogen receptor beta gene +1730 G/A polymorphism in women with polycystic ovary syndrome. AB - OBJECTIVE: To investigate whether the +1730 G/A polymorphism in the estrogen receptor (ER)-beta gene is associated with susceptibility to polycystic ovary syndrome (PCOS). DESIGN: Case-control study. SETTING: University Department of Obstetrics and Gynecology. PATIENT(S): Women with (n = 138) or without (n = 290) PCOS. INTERVENTION(S): Genotyping was performed by polymerase chain reaction restriction fragment-length polymorphism analysis. MAIN OUTCOME MEASURE(S): Genotype distribution and allele frequency of the +1730 G/A polymorphism in the ER-beta gene. RESULT(S): There was a significant difference in the genotype distribution between the patients with PCOS and controls (non-GG rates were 22.1% for patients with PCOS and 36.6% for controls). There was also a significant difference in the G and A allele frequencies between these two groups (11.7% in patients vs. 19.1% in controls with A allele). But in patients with PCOS there were no significant differences in the serum levels of hormones, biochemical variables, or ovarian morphology between GG and non-GG genotypes. CONCLUSION(S): The ER-beta gene +1730 G/A polymorphism may be associated with pathophysiologic aberrancies involved in PCOS. PMID- 19185862 TI - Factors associated with recurrence of venous leg ulcers: a survey and retrospective chart review. AB - BACKGROUND: Chronic venous leg ulcers have a significant impact on older individuals' well-being and health care resources. Unfortunately after healing, up to 70% recur. OBJECTIVE: To examine the relationships between leg ulcer recurrence and physical activity, compression, nutrition, health, psychosocial indicators and self-care activities in order to provide information for preventive strategies. DESIGN: Survey and retrospective chart review. SETTINGS: Two metropolitan hospital and three community-based leg ulcer clinics. SUBJECTS: A sample of 122 community living patients with leg ulcer of venous aetiology which had healed between 12 and 36 months prior to the survey. METHODS: Data were collected from medical records on demographics, medical history and previous ulcer history and treatments; and from self-report questionnaires on physical activity, nutrition, psychosocial measures, ulcer recurrences and history, compression and other self-care activities. All variables clinically or statistically significantly associated with recurrence at the bivariate level were entered into a logistic regression model to determine their independent influences on recurrence. RESULTS: Median follow-up time was 24 months (range 12 40 months). Sixty-eight percent of participants had recurred. Bivariate analysis found recurrence was positively associated with ulcer duration, cardiac disease, a body mass index< or =20, scoring as at risk of malnutrition and depression; and negatively associated with increased physical activity, leg elevation, wearing Class 2 (20-25 mmHg) or Class 3 (30-40 mmHg) compression hosiery, and higher self efficacy scores. After adjusting for all variables, an h/day of leg elevation (OR=0.04, 95% CI=0.01-0.17), days/week in Class 2 or 3 compression hosiery (OR=0.53, 95% CI=0.34-0.81), Yale Physical Activity Survey score (OR=0.95, 95% CI=0.92-0.98), cardiac disease (OR=5.03, 95% CI=1.01-24.93) and General Self efficacy scores (OR=0.83, 95% CI=0.72-0.94) remained significantly associated (p<0.05) with recurrence. CONCLUSIONS: Results indicate a history of cardiac disease is a risk factor for recurrence; while leg elevation, physical activity, compression hosiery and strategies to improve self-efficacy are likely to prevent recurrence. PMID- 19185863 TI - Convergence of self-reports and coworker reports of counterproductive work behavior: a cross-sectional multi-source survey among health care workers. AB - BACKGROUND: Most studies of counterproductive work behavior (CWB) are criticized for overreliance on single-source self-reports. This study attempts to triangulate on behaviors and perceptions of the work environment by linking job incumbent self-report with coworker report of the job incumbent's behaviors. Theoretical framework is the Demand-Induced Strain Compensation (DISC) Model, which proposes in general that specific job resources should match specific job demands to reduce deviant behavioral outcomes such as CWB. OBJECTIVES: To test the extent to which job incumbent self-report and coworker report of CWB in health care work converge, and the extent to which job incumbent-reported work related antecedents (i.e., job demands and job resources) similarly predict both self-reported and coworker-reported behaviors (in line with DISC theory). DESIGN: A cross-sectional survey with anonymous questionnaires was conducted, using data from two different sources (self-reports and coworker reports). SETTINGS: A large organization for residential elderly care in the Northern urban area in The Netherlands. PARTICIPANTS: Self-report and coworker questionnaires were distributed to 123 health care workers, of which 73 people returned the self report questionnaire (59% response rate). In addition, 66 out of 123 coworker questionnaires were returned (54% coworker response rate). In total 54 surveys of job incumbents and coworkers could be matched. METHODS: Next to descriptive statistics, t-test, and correlations, hierarchical regression analyses were conducted using SPSS 15.0 for Windows. RESULTS: Correlations and a t-test demonstrated significant convergence between job incumbent and coworker reports of CWB. Hierarchical regression analyses showed that both job incumbent and coworker data consistently demonstrated CWB to be related to its work-related antecedents. Specifically, findings showed that both physical and emotional job resources moderated the relation between physical job demands and CWB. CONCLUSIONS: The current findings provide stronger evidence that (multi-source measured) CWB is associated with job demands and job resources than has been provided in the past. Moreover, the present study implies that DISC theory has the potential of making a profound contribution to our understanding of counterproductive working behaviors in health care work. Future longitudinal studies should investigate this kind of relations more intensely. PMID- 19185865 TI - Increased serum levels of methylglyoxal-derived hydroimidazolone-AGE are associated with increased cardiovascular disease mortality in nondiabetic women. AB - OBJECTIVE: To investigate the association of the levels of methylglyoxal-derived hydroimidazolone AGE modified proteins (MG-H1-AGE) with cardiovascular disease (CVD) mortality in an 18-year follow-up study in Finnish nondiabetic and diabetic subjects. METHODS: The study design was a nested case-control study. Serum MG-H1 AGE levels in samples drawn at baseline were measured with a DELFIA type immunoassay in 220 diabetic subjects and 61 nondiabetic subjects who died from CVD during the follow-up, and age- and gender-matched 157 diabetic subjects and 159 nondiabetic subjects who did not die from CVD. RESULTS: In type 2 diabetic subjects serum MG-H1-AGE levels were similar in subjects who died from CVD and in subjects who did not, 32.6 (24.6-42.1) (median (interquartile range)) vs. 31.3 (22.5-40.7)U/mL (p=0.281). In nondiabetic subjects serum MG-H1 levels were significantly higher in subjects who died from CVD than in subjects who were alive, 35.4 (28.1-44.7) vs. 31.3 (24.2-38.6)U/mL (p=0.025). Corresponding MG-H1 levels were 41.2 (35.6-58.7) vs. 31.1 (26.7-35.7)U/mL, p=0.003, in women, and 34.4 (26.3-41.2) vs. 32.0 (22.8-40.3)U/mL, p=0.270, in men. Multivariate logistic regression analysis showed a significant association of serum levels of MG-H1-AGE with CVD mortality in nondiabetic women (adjusted p=0.021), but not in nondiabetic men. CONCLUSIONS: Our 18-year follow-up study shows that high baseline serum levels of MG-H1 type of AGE modified proteins were associated with CVD mortality in nondiabetic women, but not in nondiabetic men or in diabetic subjects. PMID- 19185864 TI - Association of apolipoprotein A5 concentration with serum insulin and triglyceride levels and coronary artery disease in Korean men. AB - OBJECTIVE: Whereas the relation between apolipoprotein A5 (APOA5) gene polymorphisms and triglycerides (TG) levels is well established, the associations between apoA5 concentrations, TG and coronary artery disease (CAD) remain controversial. Therefore, we investigated these relations in the setting of a case-control study involving Korean males. METHODS: ApoA5, TG, insulin, free fatty acid (FFA) and lipoprotein profiles were determined using a cross-sectional design in 777 healthy controls and 367 CAD patients. RESULTS: Plasma apoA5 concentration was lower in CAD patients than controls (192.7+/-5.2 vs. 237.2+/ 3.7ng/ml, P<0.001). Values in the second and top tertiles of apoA5 were associated with a decreased odds ratio (OR) for CAD when compared with values in the bottom tertile; OR for apoA5 top tertile was 0.33 (95% CI, 0.23-0.47) in the age- and BMI-adjusted model and 0.35 (95% CI, 0.23-0.56) following additional adjustments for smoking, drinking status, blood pressure, TG and HDL-cholesterol. After adjustment for age and BMI, plasma apoA5 concentration was negatively correlated with serum TG (r=-0.188, P<0.001) and insulin (r=-0.185, P<0.001) in normotriglyceridemic controls (TG<150mg/dL, n=509) whereas apoA5 was positively correlated with serum TG in hypertriglyceridemic controls (TG> or =150mg/dL, n=268) (r=0.246, P<0.001) and total CAD patients (r=0.177, P<0.01). Regardless of TG levels and CAD status, apoA5 concentration was positively correlated with HDL cholesterol and FFA levels. CONCLUSIONS: Our data supports an inverse association between plasma apoA5 concentrations and CAD risk, probably due to the observed negative correlations of apoA5 with TGs and insulin, although these correlations were affected by TG levels. PMID- 19185866 TI - Lipophilicity data for some preservatives estimated by reversed-phase liquid chromatography and different computation methods. AB - The chromatographic behavior of some preservatives was performed by reversed phase high-performance liquid chromatography on C18 (LiChroCART, Purosphere RP 18e), C8 (Zorbax, Eclipse XDB-C8), CN100 (Saulentechnik, Lichrosphere) and NH(2) (Supelcosil LC-NH(2)) columns. The lipophilicity estimated for the first time on the first three columns are comparable and very well correlated. The mobile phase was a mixture of methanol-water (0.1% formic acid) in different volume proportions from 40% to 60% (v/v) for RP-C18, RP-C8 and RP-CN100 column (exception for parabens on RP-C8 column-the methanol concentrations being from 55% to 65%) and from 30% to 50% (v/v) for RP-NH(2). Highly significant correlations were obtained between different experimental indices of lipophilicity (logk(w), S, phi(0), mean of k and logk, and scores of k and logk corresponding to the first principal component) and computed logP values, and C8 column seems to be more suited for estimating the lipophilicity of the investigated compounds. These direct correlations offer a very good opportunity to derive powerful predictive models via Collander-type equations. The reliability of scores values as lipophilic indices is shown by their high correlation with the logK(ow) obtained using classical "shake-flask" technique, logk(w) and also some of the computed logP values. In addition, the results obtained applying PCA to the retention data may be used in interpreting the molecular mechanism of interactions between eluents and stationary phases with different polarity and to explain the chromatographic behavior of compounds. Finally, the "congeneric lipophilicity chart" described by the scores corresponding to the first principal component has the effect of separating compounds from each other more effectively from congeneric ((dis)similarity) point of view. The parabens and tert-butylhydroquinone appeared to be the most lipophilic preservatives. PMID- 19185867 TI - Automated polyvinylidene difluoride hollow fiber liquid-phase microextraction of flunitrazepam in plasma and urine samples for gas chromatography/tandem mass spectrometry. AB - A new polyvinylidene difluoride (PVDF) hollow fiber (200 microm wall thickness, 1.2mm internal diameter, 0.2 microm pore size) was compared with two other polypropylene (PP) hollow fibers (200, 300 microm wall thickness, 1.2mm internal diameter, 0.2 microm pore size) in the automated hollow fiber liquid-phase microextraction (HF-LPME) of flunitrazepam (FLNZ) in biological samples. With higher porosity and better solvent compatibility, the PVDF hollow fiber showed advantages with faster extraction efficiency and operational accuracy. Parameters of the CTC autosampler program for HF-LPME in plasma and urine samples were carefully investigated to ensure accuracy and reproducibility. Several parameters influencing the efficiency of HF-LPME of FLNZ in plasma and urine samples were optimized, including type of porous hollow fiber, organic solvent, agitation rate, extraction time, salt concentration, organic modifier, and pH. Under optimal conditions, extraction recoveries of FLNZ in plasma and urine samples were 6.5% and 83.5%, respectively, corresponding to the enrichment factor of 13 in plasma matrix and 167 in urine matrix. Excellent sample clean-up was observed and good linearities (r(2)=0.9979 for plasma sample and 0.9995 for urine sample) were obtained in the range of 0.1-1000 ng/mL (plasma sample) and 0.01-1000 ng/mL (urine sample). The limits of detection (S/N=3) were 0.025 ng/mL in plasma matrix and 0.001 ng/mL in urine matrix by gas chromatography/mass spectrometry/mass spectrometry. PMID- 19185868 TI - Dispersive solid-phase microextraction method for sample extraction in the analysis of four tetracyclines in water and milk samples by high-performance liquid chromatography with diode-array detection. AB - A dispersive solid-phase microextraction (dispersive-SPME) method for the determination of tetracycline, oxytetracycline, chlortetracycline and doxycycline is proposed. Different silica-based and polymeric sorbents were evaluated for their capacity to simultaneously preconcentrate tetracyclines (TCs) in the dispersive format from aqueous or organic solutions. Silica-based sorbents especially functionalized with primary amine, secondary amine, or carbonyl groups have showed higher capacity than polymeric sorbents under organic environment. In the proposed dispersive solid-phase microextraction method, after extraction with acetonitrile and salt-promoted partitioning, TCs were adsorbed to a small amount of dispersive silica-based primary and secondary amine sorbents, desorbed with a small volume of desorption solution, and determined by high-performance liquid chromatography with diode-array detection. Under the optimal conditions, recoveries were determined for surface water and milk samples spiked at 10 ng/mL and 50-150 ng/g, respectively, and quantification was achieved by matrix-matched calibration. The calibration curves of four TCs in both samples showed linearity with a correlation coefficient value above 0.997. Average recoveries ranged from 97.1 to 104.1% and the precision was from 2.0 to 5.6%. Limits of detection ranged from 0.7 to 3.5 ng/mL and from 7.9 to 35.3 ng/g for four TCs surface in surface water and milk samples, respectively. PMID- 19185869 TI - Development and validation of a liquid chromatographic method for the analysis of capreomycin sulfate and its related substances. AB - A gradient LC method for the analysis of capreomycin sulfate and its related substances was developed. The chromatographic conditions include the use of a Hypersil base deactivated C(18) (250 mm x 4.6mm, 5 microm) column maintained at 25 degrees C, a mobile phase containing acetonitrile, phosphate buffer pH 2.3 and 0.025M hexanesulfonate at a flow rate of 1.0 mL/min and UV detection performed at 268 nm. Good separation of the four active components of capreomycin and eleven unknown impurities was achieved. A system suitability test to check the quality of the separation is specified. The method shows good repeatability, linearity and robustness. PMID- 19185870 TI - Characterization of sodium laureth sulfate by reversed-phase liquid chromatography with evaporative light scattering detection and 1H nuclear magnetic resonance spectroscopy. AB - A direct and effective method utilizing reversed-phase liquid chromatography combined with evaporative light scattering detection was developed to determine the relative ratio of different alkyl chain lengths, to quantify the average ethylene oxide (EO) level and to identify EO distribution in the presence of sodium laureth sulfates. A C8 bonded silica gel column and an acetonitrile-water gradient mobile phase containing ammonium acetate were used as the best stationary and mobile phase, respectively. The results were confirmed by nuclear magnetic resonance spectroscopy. The detection limit was 80 microg/mL and the calibration curve, i.e., the log-log plots (peak area vs. concentration), was linear in the working range of 80-4200 microg/mL with R(2) values of above 0.999 (in the case of 3 mol sodium laureth sulfates). Furthermore, the application of the chromatographic method to a commercial product without pretreatment was presented. The raw material was identified by fast atom bombardment mass spectrometry. PMID- 19185871 TI - Analysis of the stable free radical scavenging capability of artificial polyphenol mixtures and plant extracts by capillary electrophoresis and liquid chromatography-diode array detection-tandem mass spectrometry. AB - The oxidation process of phenolic compounds of an artificial mixture consisting of six polyphenols and the extract of eggplant (Solanum melongena) skin was monitored by using capillary zone electrophoresis and liquid chromatography-diode array detection-tandem mass spectrometry. The methods developed enabled simultaneous evaluation of the antioxidative capability of each compound. The above oxidation process was carried out using two radicals, viz. the 2,2-diphenyl 1-picrylhydrazyl and hydroxyl radicals generated via the Fenton reaction. The radical scavenging effects of artificial and natural polyphenol mixtures were compared. PMID- 19185872 TI - Rapid analysis of Radix puerariae by near-infrared spectroscopy. AB - A new, rapid analytical method using near-infrared spectroscopy (NIRS) was developed to differentiate two species of Radix puerariae (GG), Pueraria lobata (YG) and Pueraria thomsonii (FG), and to determine the contents of puerarin, daidzin and total isoflavonoid in the samples. Five isoflavonoids, puerarin, daidzin, daidzein, genistin and genistein were analyzed simultaneously by high performance liquid chromatography-diode array detection (HPLC-DAD). The total isoflavonoid content was exploited as critical parameter for successful discrimination of the two species. Scattering effect and baseline shift in the NIR spectra were corrected and the spectral features were enhanced by several pre processing methods. By using linear discriminant analysis (LDA) and soft independent modeling class analogy (SIMCA), samples were separated successfully into two different clusters corresponding to the two GG species. Furthermore, sensitivity and specificity of the classification models were determined to evaluate the performance. Finally, partial least squares (PLS) regression was used to build the correlation models. The results showed that the correlation coefficients of the prediction models are R=0.970 for the puerarin, R=0.939 for daidzin and R=0.969 for total isoflavonoid. The outcome showed that NIRS can serve as routine screening in the quality control of Chinese herbal medicine (CHM). PMID- 19185873 TI - Enantioseparation of a novel "click" chemistry derived native beta-cyclodextrin chiral stationary phase for high-performance liquid chromatography. AB - A novel native beta-cyclodextrin chiral stationary phase was prepared via "click" chemistry with cuprous iodide-triphenylphosphine complex as the catalyst and applied for enantioseparation of Dns-amino acids, substituted phenyl and phenoxy group modified propionic acids, flavonoids, and some pharmaceutical compounds such as nimodipine, propranolol, brompheniramine and bendroflumethiazide in reversed-phase high-performance liquid chromatography. The studied analytes could be resolved under different separation conditions. The resolution of Dns-DL-Leu could reach 5.08 using a mobile phase consisting of 1% (w/w) triethylammonium acetate buffer (pH 4.11) and methanol (50:50 v/v). The effects of buffer pH and the content of organic modifier on enantioseparation of Dns-amino acids by this novel chiral phase were being investigated. The separation results demonstrate that click chemistry, a versatile reaction, affords a facile approach towards the preparation of stable chiral stationary phases. PMID- 19185874 TI - Reactions of lysine with montmorillonite at 80 degrees C: implications for optical activity, H+ transfer and lysine-montmorillonite binding. AB - Amino acid-smectite interaction may have catalyzed prebiotic reactions essential for the emergence of life. Lysine solutions (0.05 M) were reacted with Na smectite in adsorption-desorption experiments. The lysine-smectite complexes were heated at 80 degrees C for 10 days to investigate (1) possible slow processes taking place at surface temperature that would be accelerated at higher temperature and (2) processes taking place in hydrothermal systems. Three sets of experiments were performed: thermal treatment in closed tubes and water added regularly; thermal treatment in closed tubes without adding water; and thermal treatment in open tubes and no added water. After lysine desorption (displacement with 0.1 M CaCl(2)), the solutions were investigated using circular dichroism (CD) and the smectite samples using FTIR and CHN elemental analysis. CD spectra were dependent on the solution pH, which was controlled by lysine protonation state. The lysine protonation state was altered by the adsorption-desorption process, with a higher Lys(+)/Lys(+/-) ratio after desorption. The CD and CHN analyses show that the thermal treatment in a moist state causes stronger smectite-lysine binding. FTIR data suggest that the stronger binding is caused by more or stronger H bonds between -NH(3)(+) lysine groups and smectite basal O atoms. PMID- 19185875 TI - Characterization of floc size, strength and structure in various aluminum coagulants treatment. AB - Flocs generated in various aluminum coagulants treatment exhibit difference in size, strength and structure. The formation, breakage and re-formation of flocs were investigated by using a laser diffraction instrument (Malvern Mastersizer 2000, Malvern, UK). Different Al coagulants--conventional Al salt (AlCl(3)), polyaluminum chloride (PAC-1) and the purified polyaluminum chloride (PAC-2)- were investigated for the coagulation of humic acid (HA). The results showed the properties of HA flocs depend on which species of aluminum used. The monomeric and dimeric aluminum species (denoted as Al(mono)) formed the largest and strongest HA flocs with longest growth time. The HA flocs formed by Al(13)O(4)(OH)(24)(7+) (Al(13) for short) is the smallest and weakest. The properties of HA flocs made it possible to investigate the relationship between the flocs growth process and aluminum species. PMID- 19185876 TI - Textural characterizations and catalytic properties of quasispherical nanosized molybdenum disulfide. AB - Synthesis of sphere nanostructured MoS(2) is reported. Characterization of the synthesized MoS(2) was investigated by X-ray diffraction, nitrogen adsorption, scanning electron microscopy (SEM) and transmission electron microscopy (TEM). It was found that the obtained MoS(2) is composed of layers that bend to form mostly spheres, with an average diameter of approximately 180 nm. The growth in crystallinity is mainly due to the increased number of the round-stacked layers of MoS(2). The catalytic activity and selectivity of the synthesized nanostructured MoS(2) for the dibenzothiophene hydrodesulfurization were investigated. The closed-circle MoS(2) layers exhibited a high selectivity for the direct sulfur removal. PMID- 19185877 TI - NiAg catalysts prepared by reduction of Ni2+ ions in aqueous hydrazine II. Support effect. AB - A series of bimetallic NiAg (Ni + Ag = 1% wt) catalysts supported on amorphous silica was synthesized via chemical reduction using hydrazine as the reducing agent at 353 K. Catalysts were prepared via impregnation or precipitation technique. It was found that the reduction of the Ni(2+) ions occurred only in the presence of silver, otherwise a stable blue [Ni(N(2)H(4))(3)](2+) complex was formed. Comparisons with similar NiAg catalysts supported on crystallized silica as prepared in our previous work indicated that the Ni(2+) ions weakly interacted with acidic crystallized silica on which they were readily reduced. For both supports, the combination of silver and nickel gave rise to a synergistic effect due to the existence of NiAg groupings. The surface and catalytic properties of the metal particles formed depended on the Ni:Ag ratio, method of preparation, and acidity of the support. PMID- 19185878 TI - Dendrimer-encapsulated Pt nanoparticles in supercritical medium: synthesis, characterization, and application to device fabrication. AB - In this work we describe a general method for formation of Pt nanoparticles within an ultrathin film matrix and its application for non-volatile memory (NVM). Our approach involves the formation of Pt nanoparticles within ultrathin film matrix formed by covalent layer-by-layer (LbL) assembly of pyromellitic dianhydride (PMDA) and second generation of polyamidoamine (PAMAM) dendrimer in supercritical carbon dioxide (SCCO(2)). The hyperbranched component in the film structure serves to confine nanoparticle size and improve distribution. The memory effect and retention capability is demonstrated by means of a metal insulator semiconductor (MIS) device fabricated using the nanoparticle-laden thin film as the insulating layer. PMID- 19185879 TI - Synthesis of mesoporous zeolite Ni-MFI with high nickel contents by using the ionic complex [(C4H9)4N]2(+)[Ni(EDTA)]2- as a template. AB - Ni-MFI zeolites with high percentage of Ni (5-15 wt%) were prepared by using an ionic complex [(C(4)H(9))(4)N](2)(+)[Ni(EDTA)](2-) by one step synthesis. These molecular sieves were characterized using X-ray diffraction (XRD), X-ray fluorescence (XRF), scanning electron microscopy (SEM), high-resolution transmission electron microscopy (HRTEM), Fourier-transform infrared spectroscopy (FT-IR), ultraviolet-visible spectroscopy (UV-vis), differential scanning calorimeter (DSC) and nitrogen adsorption-desorption isotherms. The results showed that heteroatom Ni was successfully introduced into the MFI framework up to 15 wt%. Moreover, this Ni-MFI possessed regular and stable structure with high specific surface area and average pore diameter of 388-439 m(2) g(-1) and 2.566 3.828 nm, respectively, compared to MFI prepared by traditional methods. These samples also showed good thermal and hydrothermal stability. PMID- 19185880 TI - On the effect of pH on spreading of surfactant solutions on hydrophobic surfaces. AB - Surfactants are invaluable in a number of agricultural applications in products such as pesticides and herbicides. In these products, surfactants are very often used in conjunction with acidifiers in order to improve their half-life. In this paper, we investigate how the change in pH affects surfactant wetting and spreading. We compare the performance of a conventional surfactant, Triton X-100, with that of a trisiloxane superspreader, Silwet L-77, on a number of polymer coated surfaces exhibiting various degrees of hydrophobicity. Silwet L-77 in water based solutions showed very good wetting capability on all surfaces. However, its wetting ability was drastically reduced with the addition of acetic acid. On the other hand, Triton X-100 was not affected by the addition of acid and exhibited the same spreading behaviour as in water-based solutions. PMID- 19185881 TI - Profiles of dysfluency and errors in classroom discourse among children with language impairment. AB - The current study is part of a larger study that focused on within-group classroom discourse patterns of children in special education classes for language impairment (LI). Eleven children with LI aged 7-9 were examined across 4 classroom contexts: dialogic journal-writing, group lesson, peer play and sharing time. Amount and complexity of language produced (mean length of turn, type token ratio), dysfluency (self-correction, self-repetition and speaker overlap) and communicative errors were plotted for visual analysis of profiles of performance. Results showed group differences in amount and complexity of language produced, dysfluency and errors. Profiles revealed individual variation in patterns of performance that tended to cluster into sub-group similarities in amount and complexity of talk, dysfluency and errors. Implications relate to the sub classification of LI, as well as to assessment of and therapy for such subgroups. LEARNING OUTCOMES: The reader will be able to consider why within-group patterns offer different and more qualitative information than cross-group comparisons, and understand the need for both approaches to the study of language in atypically developing children. The reader will be able to differentiate between group patterns and individual differences in dimensions of classroom discourse. The reader will be able to appreciate the impact of social context on different dimensions of communication among children with LI in the classroom. The reader will be able to discuss the role that individual differences and group patterns bear on issues of assessment and treatment of LI in the school system. PMID- 19185882 TI - Increasing number of stroke specialists should contribute to utilization of IV rt PA: results of questionnaires from 1,466 hospitals in Japan. AB - PURPOSE: To determine the present status of intravenous recombinant tissue plasminogen activator (IV rt-PA) administration in Japan, we investigated the components of stroke case related to IV rt-PA utilization using a questionnaire sent to hospitals. METHODS: Questionnaires about the infrastructure of acute stroke care were sent to 8,589 hospitals between August and October 2007. Responses were categorized as follows: 1) stroke service run by stroke physicians (SPs) 24 h/day, 7 days/week (24/7); 2) IV rt-PA utilizable 24/7 (rt-PA hospitals); 3) the total number of SPs. The components related to rt-PA hospitals were analyzed and the significance of the number in SPs to the rt-PA hospital was investigated. RESULTS: Responses were received from 4,690 (54.7%) of 8,569 hospitals. Of these, 1,466 hospitals were admitting acute stroke patients. 519 of those hospitals were rt-PA hospitals. Of the 1,466 (35.4%), 48.4% were serviced 24/7 by SPs, with 75.2% having <5 SPs. Multivariate analysis revealed administration of rt-PA was significantly associated with >4 SPs (odds ratios (OR), 2.8; 95% confidence interval (95%CI), 1.9-4.1; p<0.001). Compared to hospitals with 0-1 SPs as a reference, the OR for rt-PA utilization was 5.6 (95%CI, 2.5-12.9; p<0.001) with 5 SPs, 10.8 (95%CI, 5.0-23.6; p<0.001) with 6-10 SPs, and 37.3 (95%CI, 6.5-213.1; p<0.001) with >10 SPs. CONCLUSIONS: An increased number of SPs was associated with increased IV rt-PA utilization. Development of stroke centers with larger numbers of SPs is therefore urgently needed. PMID- 19185883 TI - Do patients with amyotrophic lateral sclerosis (ALS) have increased energy needs? AB - BACKGROUND AND AIMS: Nutritional status is a prognostic factor for survival in amyotrophic lateral sclerosis (ALS) patients. We investigated the contribution of some of the components contributing to resting energy expenditure (REE) in order to determine whether potentially higher energy needs should be considered for these patients. METHODS: Thirty three ALS patients and 33 age- and gender-matched healthy controls participated. REE was measured by an open-circuit indirect calorimeter, body composition by dual energy X-ray absorptiometry, and estimated caloric intake by 7-day food records. RESULTS: Patients had lower body mass indices and lower lean body mass (LBM) than healthy controls. REE values (as a percentage of predicted) was similar but increased when normalized by LBM (P<0.001). LBM and REE decreased while REE/LBM increased in ten patients who were reassessed 6 months later. A model for predicting measured REE was constructed based on the different components, with 86% prediction of its variability. CONCLUSIONS: ALS is associated with increased REE. Various factors, such as poor caloric intake and mechanical ventilation, may mask this tendency. All the above parameters need to be considered during nutritional intervention to prevent additional muscle loss. PMID- 19185884 TI - C-glycosylanthocyanidins synthesized from C-glycosylflavones. AB - Nine C-glycosyldeoxyanthocyanidins, 6-C-beta-glucopyranosyl-7-O methylapigeninidin, 6-C-beta-glucopyranosyl-7-O-methylluteolinidin, 6-C-beta-(2'' O-beta-glucopyranosylglucopyranosyl)-7-O-methylapigeninidin, 6-C-beta-(2''-O-beta glucopyranosylglucopyranosyl)-7,4'-di-O-methylapigeninidin, 8-C-beta glucopyranosylapigeninidin, 8-C-beta-(2''-O-alpha rhamnopyranosylglucopyranosyl)apigeninidin, 8-C-beta-(2''-O-alpha-(4'''-O acetylrhamnopyranosyl)glucopyranosyl)apigeninidin, 6,8-di-C-beta glucopyranosylapigeninidin (8), 6,8-di-C-beta-glucopyranosyl-4'-O methylluteolinidin (9), have been synthesized from their respective C glycosylflavones (yields between 14% and 32%) by the Clemmensen reduction reaction using zinc-amalgam. The various precursors (C-glycosylflavones) of the C glycosylanthocyanidins were isolated from either flowers of Iris sibirica L., leaves of Hawthorn 'Crataegi Folium Cum Flore', or lemons and oranges. This is the first time C-glycosylanthocyanidins have been synthesized. The structures of all flavonoids including the flavone rotamers were elucidated by 2D NMR techniques and high-resolution electrospray MS. The distribution of the various structural forms of 8 and 9 are different at pH 1.1, 4.5, and 7.0, however, the two pigments undergoes similar structural transformations at the various pH values. Pigments 8 and 9 with C-C linkages between the sugar moieties and the aglycone, were found to be far more stable towards acid hydrolysis than pelargonidin 3-O-glucoside, which has the typical anthocyanidin C-O linkage between the sugar and aglycone. This stability may extend the present use of anthocyanins as nutraceuticals, pharmaceuticals or colorants. PMID- 19185885 TI - Impact assessment of the use of insecticide-treated bed nets on parasitaemia and anaemia for malaria control in children, Ogun State, Nigeria. PMID- 19185886 TI - Public health centres in contemporary Japan. PMID- 19185887 TI - Patterns and predictors of re-admission to hospital with self-poisoning in Scotland. AB - OBJECTIVES: To identify factors influencing hospital re-admission with self poisoning. STUDY DESIGN: Retrospective cohort follow-up study using national linked hospital discharge data. METHODS: All Scottish adult hospital episodes with self-poisoning admissions were captured using NHS Scotland Information Services Division data, and first-time 'index' admissions between 1996 and 2002 were identified. Re-admission rate was defined as the proportion of index admissions who went on to have one or more further self-poisoning admissions within 2 years. The effects of various potential risk factors for re-admission were examined using logistic regression. RESULTS: In total, 50,891 index admissions were identified; of these, 8278 patients were re-admitted. The 1-year re-admission rate was 12.2%. Older patients (>65 years) were least likely to be re-admitted [odds ratio (OR) 0.40, P<0.01, compared with patients aged 15-24 years]. No differences were found between males and females. Previous psychiatric hospital admission was associated with an increased re-admission rate (OR 2.85, P<0.01), with a diagnosis of personality disorder associated with the highest rate of re-admission (OR 4.59, P<0.01). Other factors predicting re-admission were: increased deprivation (quintile 3: OR 1.16, P<0.01; quintile 5: OR 1.15, P<0.01, compared with quintile 1); taking medicines for chronic disease, drug dependency (OR 1.6 and 1.19, P < or = 0.02) or antidepressants (OR 1.11, P=0.01) (compared with paracetamol); and co-ingestion of three or more agents (OR 1.37, P<0.01). CONCLUSION: Younger age, higher deprivation, ingestion of certain drug groups or multiple drug types, and prior psychiatric hospital admission are all risk factors for re-admission with self-poisoning. Personality disorder carried the greatest risk of re-admission. These findings may provide a basis to develop policies to reduce re-admission rates in the future. PMID- 19185888 TI - Decreasing number of public health centres (hokenjyo) in Japan. PMID- 19185890 TI - Alcohol consumption and attitudes towards banning alcohol sales on campus among European university students. AB - OBJECTIVES: The European Commission's new health strategy for improving health at the European Union (EU) level includes tackling alcohol consumption. This study aimed to assess the prevalence of alcohol consumption and problem drinking, as well as students' attitudes towards banning the sale of alcohol on campus. STUDY DESIGN: In total, 5826 students from universities in seven European countries (Denmark, Germany, Spain, Lithuania, Poland, Bulgaria and Turkey) took part in this cross-sectional study. METHODS: A self-administered questionnaire assessed sociodemographic information, frequency of alcohol consumption, problem drinking and attitudes towards banning the sale of alcohol on campus. RESULTS: The highest prevalence of drinking alcohol more than once per week was reported in Bulgarian (males 46%, females 64%) and Spanish students (males 59%, females 64%). Among those students who drank alcohol (n=3170), problem drinking (CAGE score >1) was found in 24% of males and 13% of females. Male gender, depressive moods and a low importance of good grades at university were risk factors for drinking alcohol more than once per week as well as for problem drinking. There were substantial country differences in the proportion of students who would support a ban of alcohol sales on campus (23% in Denmark, 88% in Poland). Support for a ban was higher among female students and among students who drank alcohol once or less per week. CONCLUSIONS: Problem drinking is a concern among students in many European countries, especially among males. Students' support for banning the sale of alcohol on campus varies between countries and should be considered in developing EU policy. PMID- 19185891 TI - Differential expression of cell cycle regulators p21, p27 and p53 in metastasizing canine mammary adenocarcinomas versus normal mammary glands. AB - The cyclin dependent kinase inhibitors p21 and p27 are important regulators of cell cycle progression. To analyze their role in the malignant progression of canine mammary tumors expression levels of p27 and p21 and its major regulator p53 were compared in simple adenomas, adenocarcinomas of the mammary gland and lymph node metastases with normal mammary gland. Laser microdissection of tissue samples and real-time PCR were used for quantification of mRNA expression levels. p21 was overexpressed in adenocarcinomas, whereas adenomas and metastases expressed p21 more heterogeneously. Comparison of p21 expression in adenocarcinomas and their metastases revealed a significant decrease in expression in metastases. In contrast, p27 expression was reduced in the adenocarcinomas but heterogeneously expressed in adenomas and metastases. Taken together the results suggest that loss of p21 overexpression is associated with tumor metastasis while reduced cell cycle inhibition by p27 is associated with malignant progression. PMID- 19185892 TI - Detection of porcine Circovirus type 2 (PCV2) variants PCV2-1 and PCV2-2 in Brazilian pig population. AB - In the present study whole genome of six Brazilian isolates of PCV2 were sequenced, analyzed and compared with 35 other sequences (24 from other countries and 17 from Brazil). The phylogenetic analysis showed that mostly Brazilian variants of PCV2 were grouped as PCV2-1. Two isolates among the six analyzed here could not be grouped with any other PCV2-2 analyzed in this study. One of these isolates was from an aborted fetus with myocarditis and the other from a PMWS affected pig. The results pointed here showed that both groups of PCV2 are present in Brazilian pig population without any clear geographical correlation. PMID- 19185893 TI - Effectiveness of small interfering RNA (siRNA) against the Mcl-1 gene in a canine mammary gland tumor cell line. AB - The effect of down-regulation of Mcl-1 expression by small interfering RNA (siRNA) against the canine Mcl-1 gene on apoptosis was investigated by transfecting CF33 (canine mammary gland tumor cell line) with siRNA using cationic liposomes. The siRNA against canine Mcl-1 increased the rate of apoptotic cells and decreased the numbers of viable cells. Further, sequence specific down-regulation of Mcl-1 expression was measured by real time-PCR and Western blot analysis. The siRNA directed against the Mcl-1 gene reduced both the mRNA and protein expression in the CF33. Our study suggests the importance of Mcl 1 in canine mammary tumors for inducing apoptosis and reinforces using Mcl-1 as a putative therapeutic target in canine mammary gland tumor. PMID- 19185894 TI - Serological survey of Encephalitozoon cuniculi in farm rabbits in Italy. AB - Rabbit sera (n=1600) from 40 commercial farms were submitted to a serological screening for Encephalitozoon cuniculi by an enzyme-linked immunosorbent assay (ELISA) and a carbon immunoassay (CIA test). Antibodies anti-Encephalitozoon cuniculi were found in 505/1600 (31.6%) sera analysed, and all the farms (100%) resulted positive. Rabbits older than 4 months showed a significantly higher seropositivity for E. cuniculi (chi-squared test: p<0.0001) than rabbits under 4 months, E. cuniculi sero-prevalence showed an increasing trend in rabbits within the farm along with the increase in the "number of rabbits on the farm"; however, this trend was not significant (Spearman's correlation: p=0.073). The findings of the present study confirm that rabbit is the main reservoir of E. cuniculi; they are of epidemiological relevance and immediate public health importance because of the recognized infectivity in humans by the microsporidium. PMID- 19185895 TI - Paragonimiasis: a common cause of persistent pleural effusion in Lao PDR. AB - Southeast Asia is the major endemic area for paragonimiasis. Diagnosis relies on identification of ova in the sputum, pleural fluid or tissue specimen, or serology. Low awareness, however, frequently results in the disease being overlooked. We report nine cases presenting as primary, massive and protracted pleural effusions. All patients had evidence of Paragonimus spp. in the pleural fluid; one discharged an adult worm through a chest tube during treatment with praziquantel. In three cases, resolution of symptoms and pleural effusions could not be achieved, despite repeated fluid evacuation procedures and courses of praziquantel, which contradicts the widely accepted statement of paragonimiasis being self-limited and easy to cure. The disease should be considered in any case of elusive pleural effusion occurring in endemic areas. PMID- 19185896 TI - Isolation and genotyping of potentially pathogenic Acanthamoeba strains from dust sources in Iran. AB - Opportunistic free-living amoebae of the Acanthamoeba genus present genotypes/strains that are causative agents of Acanthamoeba keratitis (AK), with genotype T4 related to most AK cases worldwide. In order to prove that exposure to dirt and dust could be an important risk factor for contact lens wearers in Iran, 13 strains isolated from dust samples in this area were classified at the genotype level. Results revealed the presence of T4, T5 and T11 genotypes within these samples. To our knowledge, this is the first study presenting the identification of pathogenic genotypes of Acanthamoeba in dust samples in Iran and around the world. PMID- 19185897 TI - Socio-environmental factors and ascariasis infection among school-aged children in Ilobu, Osun State, Nigeria. AB - The risk factors predisposing children to ascariasis transmission in a rural community of Osun State, Nigeria were investigated from November 2005 to April 2006. Children below 16 years of age were examined at the household level after information on biodata, access to water supply and sanitation, socio-economic status of their parents and degree of cohabitation with their parents was collected using a questionnaire. Of 440 children examined, overall prevalence was 60% and median intensity was 1548 eggs per gram (epg) (min. 48 epg; max. 55464 epg). Infection patterns were gender comparable and age dependent, with peak prevalence (67.8%) occurring in children aged 5-9 years and peak median intensity (4368 epg) in children aged >or=15 years. Logistic regression analysis revealed that prevalence was influenced by patterns of water supply and sanitation, parents' educational background, number of biological parents living with a child and number of playmates a child has. These findings suggest that socio environmental risk factors which play a role in disease transmission need to be taken into account when formulating sustainable control strategies for ascariasis and other intestinal parasites in Nigeria and elsewhere. PMID- 19185898 TI - High rate of fatigue and abdominal symptoms 2 years after an outbreak of giardiasis. AB - The aim of this study was to evaluate the prevalence of fatigue and abdominal symptoms 2 years after Giardia lamblia infection. All 1262 cases who had Giardia positive stool samples during an outbreak in 2004 in Norway received a questionnaire in 2006 asking about fatigue and abdominal symptoms. Fatigue was reported by 41%, whereas 38% reported abdominal symptoms, and there was a highly significant association between these symptoms. Increasing age was a highly significant risk factor for fatigue. The symptoms were not due to chronic infection in this cohort. Our data warrant further investigations into the late effects of giardiasis. PMID- 19185899 TI - HIV entry in macrophages is dependent on intact lipid rafts. AB - Macrophages are an important natural target cell for HIV-1, but previous studies of virus entry into these cells are limited, and the involvement of membrane cholesterol and lipid rafts is unknown. Cholesterol disruption of macrophage membranes using four pharmacological agents acting by different mechanisms: methyl-beta cyclodextrin, nystatin, filipin complex and Lovastatin, all significantly inhibited productive HIV entry and reverse transcription. The inhibitory effects of these drugs resulted in decreased virus release from infected cells, and could be substantially reversed by the addition of water soluble cholesterol. The virus bound equally to cholesterol-disrupted cells even though HIV receptor expression levels were significantly reduced. Macrophage CD4 and CCR5 were found to partition with the detergent-resistant membranes with a typical raft-associating protein flotillin-1. HIV particles were observed co localising with a marker of lipid rafts (CTB-FITC) early post infection. These data suggest that macrophage membrane cholesterol is essential for HIV entry, and implicate lipid raft involvement. PMID- 19185900 TI - Antibiotics in the aquatic environment--a review--part I. AB - Although antibiotics have been used in large quantities for some decades, until recently the existence of these substances in the environment has received little notice. It is only in recent years that a more complex investigation of antibiotic substances has been undertaken in order to permit an assessment of the environmental risks they may pose. Within the last decade an increasing number of studies covering antibiotic input, occurrence, fate and effects have been published, but there is still a lack of understanding and knowledge about antibiotics in the aquatic environment despite the numerous studies performed. This review addresses the present state of knowledge concerning the input, occurrence, fate and effects of antibiotics in the environment. It brings up important questions that are still open, and addresses some significant issues which must be tackled in the future for a better understanding of the behavior of antibiotics in the environment, as well as the risks associated with their occurrence. Questions related to resistance in the environment that may be caused by antibiotics will be addressed in the second part. PMID- 19185901 TI - A comparative study of Saccharomyces cerevisiae sensitivity against eight yeast species sensitivities to a range of toxicants. AB - Fungi are widespread and very important in many ecosystems but the extensive use of pesticides can adversely affect beneficial fungi. The yeast, Saccharomyces cerevisiae has been proposed for the toxicological assessment of the effects of environmental pollutants on non-target fungi. To assess whether S. cerevisiae is a good representative of the immense ecological and phylogenetic diversity of yeasts, we compare the sensitivity of eight other yeasts from diverse phylogenetic taxa to a range of toxicants and environmental samples. Sensitivity was assessed using both the growth inhibition and alamar blue (resorufin fluorescence inhibition) bioassays. The growth inhibition bioassay showed that all yeast species had similar dose-response curves for the five organic fungicides and two environmental samples used. However, two yeast species, Trichosporon dulcitum and Pseudozyma fusiformata, were a great deal more sensitive than all others to CuSO4 and K2Cr2O7 while S. cerevisiae was the most tolerant to these chemicals. S. cerevisiae, however, showed similar sensitivity as other species to all toxicants in the resorufin fluorescence inhibition bioassay. It can therefore be used as a representative yeast species for assessing effects of environmental contaminants to non-target fungi and in the screening of chemical libraries for fungicidal activity. PMID- 19185902 TI - Semicircular canal system in early primates. AB - Mammals with more rapid and agile locomotion have larger semicircular canals relative to body mass than species that move more slowly. Measurements of semicircular canals in extant mammals with known locomotor behaviours can provide a basis for testing hypotheses about locomotion in fossil primates that is independent of postcranial remains, and a means of reconstructing locomotor behaviour in species known only from cranial material. Semicircular canal radii were measured using ultra high resolution X-ray CT data for 9 stem primates ("plesiadapiforms"; n=11), 7 adapoids (n=12), 4 omomyoids (n=5), and the possible omomyoid Rooneyia viejaensis (n=1). These were compared with a modern sample (210 species including 91 primates) with known locomotor behaviours. The predicted locomotor agilities for extinct primates generally follow expectations based on known postcrania for those taxa. "Plesiadapiforms" and adapids have relatively small semicircular canals, suggesting they practiced less agile locomotion than other fossil primates in the sample, which is consistent with reconstructions of them as less specialized for leaping. The derived notharctid adapoids (excluding Cantius) and all omomyoids sampled have relatively larger semicircular canals, suggesting that they were more agile, with Microchoerus in particular being reconstructed as having had very jerky locomotion with relatively high magnitude accelerations of the head. Rooneyia viejaensis is reconstructed as having been similarly agile to omomyids and derived notharctid adapoids, which suggests that when postcranial material is found for this species it will exhibit features for some leaping behaviour, or for a locomotor mode requiring a similar degree of agility. PMID- 19185903 TI - Pilot-scale comparison of constructed wetlands operated under high hydraulic loading rates and attached biofilm reactors for domestic wastewater treatment. AB - Four different pilot-scale treatment units were constructed to compare the feasibility of treating domestic wastewater in the City of Heraklio, Crete, Greece: (a) a free water surface (FWS) wetland system, (b) a horizontal subsurface flow (HSF) wetland system, (c) a rotating biological contactor (RBC), and (d) a packed bed filter (PBF). All units operated in parallel at various hydraulic loading rates (HLR) ranging from 50% to 175% of designed operating HLR. The study was conducted during an 8 month period and showed that COD removal efficiency of HSF was comparable (>75%) to that of RBC and PBF, whereas that of the FWS system was only 57%. Average nutrient removal efficiencies for FWS, HSF, RBC and PBF were 6%, 21%, 40% and 43%, respectively for total nitrogen and 21%, 39%, 41% and 42%, respectively for total phosphorus. Removals of total coliforms were lowest in FWS and PBF (1.3 log units) and higher in HSF and RBC (2.3 to 2.6 log units). HSF showed slightly lower but comparable effluent quality to that of RBC and PBF systems, but the construction cost and energy requirements for this system are significantly lower. Overall the final decision for the best non conventional wastewater treatment system depends on the construction and operation cost, the area demand and the required quality of effluent. PMID- 19185904 TI - Application of land use regression to regulatory air quality data in Japan. AB - A land use regression (LUR) model has been used successfully for predicting traffic-related pollutants, although its application has been limited to Europe and North America. Therefore, we modeled traffic-related pollutants by LUR then examined whether LUR models could be constructed using a regulatory monitoring network in Shizuoka, Japan. We used the annual-mean nitrogen dioxide (NO2) and suspended particulate matter (SPM) concentrations between April 2000 and March 2006 in the study area. SPM accounts for particulate matter with an aerodynamic diameter less than 8 microm (PM(8)). Geographic variables that are considered to predict traffic-related pollutants were classified into four groups: road type, traffic intensity, land use, and physical component. Using geographical variables, we then constructed a model to predict the monitored levels of NO2 and SPM. The mean concentrations of NO2 and SPM were 35.75 microg/m(3) (standard deviation of 11.28) and 28.67 microg/m(3) (standard deviation of 4.73), respectively. The final regression model for the NO2 concentration included five independent variables. R(2) for the NO2 model was 0.54. On the other hand, the regression model for the SPM concentration included only one independent variable. R(2) for the SPM model was quite low (R(2) = 0.11). The present study showed that even if we used regulatory monitoring air quality data, we could estimate NO2 moderately well. This result could encourage the wide use of LUR models in Asian countries. PMID- 19185905 TI - Major ion chemistry and dissolved inorganic carbon cycling in a human-disturbed mountainous river (the Luodingjiang River) of the Zhujiang (Pearl River), China. AB - Major ion chemistry and dissolved inorganic carbon system (DIC, mainly HCO3(-) and gaseous CO2) in the Luodingjiang River, a mountainous tributary of the Zhujiang (Pearl River), China, were examined based on a seasonal and spatial sampling scheme in 2005. The diverse distribution of lithology and anthropogenic impacts in the river basin provided the basic idea to assess the effects of lithology vs. human activities on water chemistry and carbon biogeochemistry in river systems. Major ions showed great spatial variations, with higher concentrations of total dissolved solids (TDS) and DIC in the regions with carbonate rocks and clastic sedimentary rocks, while lower in the regions with metamorphic sandstones and schists as well as granites. pCO2 at all sampling sites was oversaturated in June, ranging with a factor from 1.6 to 18.8 of the atmospheric concentration, reflecting the enhanced contribution from baseflow and interflow influx as well as in situ oxidation of organic matter. However, in April and December, undersaturated pCO2 was found in some shallow, clean rivers in the upstream regions. delta13C of DIC has a narrow range from -9.07 to -13.59 per thousand, which was more depleted in the regions with metamorphic rocks and granites than in the carbonate regions. Seasonally, it was slightly more depleted in the dry season (December) than in the wet season (June). The results suggested that lithological variability had a dominant control on spatial variations of water chemistry and carbon geochemistry in river systems. Besides, anthropogenic activities, such as agricultural and urban activities and in-stream damming, as well as river physical properties, such as water depth and transparency, also indicated their impacts. The seasonal variations likely reflected the changes of hydrological regime, as well as metabolic processes in the river. PMID- 19185907 TI - Protein Z g-42a variant and the risk of pregnancy-related venous thromboembolism in a cohort of Italian patients. PMID- 19185906 TI - Leukocyte count is associated with increased platelet reactivity and diminished response to aspirin in healthy individuals with a family history of coronary artery disease. AB - BACKGROUND: Markers of systemic inflammation, including blood leukocyte count, are associated with increased cardiovascular risk, but the mechanisms underlying this association are unclear. Leukocytes may promote platelet reactivity and thrombus formation, providing a basis for increased risk, but a relation between leukocyte count and platelet function has not been studied. METHODS: We evaluated the relation of blood leukocyte count, C-reactive protein (CRP), and interleukin 6 (IL-6) to platelet aggregation to collagen, ADP and arachidonic acid, and to urinary excretion of 11-dehydro thromboxane B2. Studies were conducted in 1600 individuals (45.0+/-12.9 years, 42.7% male) at risk for coronary artery disease (CAD) before and after low dose aspirin. RESULTS: At baseline, platelet reactivity increased with increasing quartile of leukocyte count (median counts for each quartile were normal) for all measures of platelet function (P<0.0001). These relations were unchanged by aspirin. The relation between leukocyte count and each measure of platelet reactivity remained significant (P<0.05) after multivariable adjustment for CRP, IL-6, cardiac risk factors, hematologic variables, and platelet thromboxane production. CRP and IL-6 were independently associated with few measures of platelet reactivity. CONCLUSIONS: Increasing quartile of leukocyte count, even within the normal range, is associated with increasing platelet reactivity in individuals at risk for CAD. This relationship is not altered by aspirin and is independent of inflammatory markers and platelet thromboxane production. Additional studies are needed to determine the mechanism(s) for this association and therapies to reduce cardiovascular risk in patients with elevated leukocyte counts. PMID- 19185908 TI - Combined androgen blockade revisited: emerging options for the treatment of castration-resistant prostate cancer. PMID- 19185909 TI - Early conception factor lateral flow assays for pregnancy in the mare. AB - The ECF lateral flow assay test is marketed to detect non-pregnancy in mares. The objectives of the present study were to determine the accuracy of the ECF test, the accuracy of the electronic reader accompanying the ECF test, and agreement between two human readers and the electronic reader. Serum samples were collected from anestrus, cycling but not inseminated, and inseminated mares, and were evaluated with the ECF test (EDP Biotech Company, Knoxville, TN, USA) at The Ohio State University and at the EDP Biotech Laboratory. Specificity ranged from 0.07 to 0.16, the negative predictive value ranged from 0.15 to 0.33, and accuracy ranged from 0.43 to 0.52. The electronic reader did not add improve the accuracy or predictive values of the test. Based on the electronic reader, 80.0% of the serum samples collected from the anestrus mares were false positives; Readers 1 and 2 had 60.0 and 33.3% false positives, respectively. For samples collected during the estrous cycle, 83.9% were false positives by the electronic reader, whereas Readers 1 and 2 had 43.7 and 26.4% false positives. We concluded that, regardless of whether the test strips were evaluated by a human or electronic reader, this assay was not accurate for determination of the non-pregnant mare. PMID- 19185910 TI - Morphological characterization and meiotic competence of oocytes collected from filly ovaries. AB - The effect of filly age on morphology of the ovaries, collected oocytes and their capacity for in vitro maturation (IVM) was examined. The ovaries of slaughtered fillies were classified into three groups, according to filly age: (I) <10 month old (<10MF); (II) approximately 1 year old (1YF); and (III) approximately 1.5 year old (1.5YF). The ovaries of mares were used as a control group. Ovarian morphology and collected oocytes were evaluated. Only oocytes with expanded (Ex) and compact (Cm) cumuli were used for IVM. In <10MF, 1YF, 1.5YF and mare groups, corpora lutea were found in the ovaries of 9.3%, 36.7%, 59.6% and 80.9% females, respectively (P<0.001). Based on this observation, we found that about 37% of fillies reach puberty at approximately 12 months of age. No relationship was found between filly age and morphology of the oocytes obtained. In comparison to mares, fewer (P<0.05) Cm oocytes were collected from filly ovaries. Among Cm groups, fewer filly (28.4-35.5%) than mare oocytes (50.0%) reached metaphase II stage, but the difference was only significant when compared to oocytes of the <10MF group (P<0.05). In the Ex groups, a similar proportion of oocytes of fillies (40.8-51.1%) and mares (48.4%) attained the metaphase II stage. In conclusion, in the culture conditions applied, Cm oocytes of fillies younger than 10 months showed lower meiotic competence than mare oocytes. Oocytes of older fillies showed meiotic competence similar (P>0.05) to mare oocytes. Both filly and mare oocytes with expanded cumuli displayed the same capacity for IVM. PMID- 19185911 TI - Structural optimization of the fibre-reinforced composite substructure in a three unit dental bridge. AB - INTRODUCTION: Failures of fixed partial dentures (FPDs) made of fibre-reinforced composites (FRC) have been reported in many clinical and in vitro studies. The types of failure include debonding at the composite-tooth interface, delamination of the veneering material from the FRC substructure and fracture of the pontic. The design of the FRC substructure, i.e. the position and orientation of the fibres, will affect the fracture resistance of the FPD. OBJECTIVES: The purpose of this study was to find an optimal arrangement of the FRC substructure, by means of structural optimization, which could minimize the failure-initiating stresses in a three-unit FPD. METHODS: A structural optimization method mimicking biological adaptive growth was developed for orthotropic materials such as FRC and incorporated into the finite element (FE) program ABAQUS. Using the program, optimization of the fibre positions and directions in a three-unit FPD was carried out, the aim being to align the fibre directions with those of the maximum principal stresses. The optimized design was then modeled and analyzed to verify the improvements in mechanical performance of the FPD. RESULTS: Results obtained from the optimization suggested that the fibres should be placed at the bottom of the pontic, forming a U-shape substructure that extended into the connectors linking the teeth and the pontic. FE analyses of the optimized design indicated stress reduction in both the veneering composite and at the interface between the veneer and the FRC substructure. SIGNIFICANCE: The optimized design obtained using FE-based structural optimization can potentially improve the fracture resistance of FPDs by reducing some of the failure-initiating stresses. Optimization methods can therefore be a useful tool to provide sound scientific guidelines for the design of FRC substructures in FPDs. PMID- 19185912 TI - Human hepatocyte functions in a crossed hollow fiber membrane bioreactor. AB - An important challenge in liver tissue engineering is the development of bioartificial systems that are able to favour the liver reconstruction and to modulate liver cell behaviour. A crossed hollow fiber membrane bioreactor was developed to support the long-term maintenance and differentiation of human hepatocytes. The bioreactor consists of two types of hollow fiber (HF) membranes with different molecular weight cut-off (MWCO) and physico-chemical properties cross-assembled in alternating manner: modified polyetheretherketone (PEEK-WC) and polyethersulfone (PES), used for the medium inflow and outflow, respectively. The combination of these two fiber set produces an extracapillary network for the adhesion of cells and a high mass exchange through the cross-flow of culture medium. The transport of liver specific products such as albumin and urea together with the transport of drug such as diazepam was modelled and compared with the experimental metabolic data. The theoretical metabolite concentration differed 7.5% for albumin and 5% for urea with respect to experimental data. The optimised perfusion conditions of the bioreactor allowed the maintenance of liver functions in terms of urea synthesis, albumin secretion and diazepam biotransformation up to 18 days of culture. In particular the good performance of the bioreactor was confirmed by the high rate of urea synthesis (28.7 microg/h 10(6) cells) and diazepam biotransformation. In the bioreactor human hepatocytes expressed at high levels the individual cytochrome P450 isoenzymes involved in the diazepam metabolism. The results demonstrated that crossed HF membrane bioreactor is able to support the maintenance of primary human hepatocytes preserving their liver specific functions for all investigated period. This device may be a potential tool in the liver tissue engineering for drug metabolism/toxicity testing and study of disease pathogenesis alternatively to animal experimentation. PMID- 19185913 TI - Nanoparticle-mediated local delivery of Methylprednisolone after spinal cord injury. AB - Systemic administration of a high-dose of Methylprednisolone (MP) can reduce neurological deficits after acute spinal cord injury (SCI). However, the use of high-dose MP in treating acute SCI is controversial due to significant dose related side effects and relatively modest improvements in neurological function. Here, using a rat model of SCI, we compare the efficacy of controlled, nanoparticle-enabled local delivery of MP to the injured spinal cord with systemic delivery of MP, and a single local injection of MP without nanoparticles. Based on histological and behavioral data, we report that local, sustained delivery of MP via nanoparticles is significantly more effective than systemic delivery. Relative to systemic delivery, MP-nanoparticle therapy significantly reduced lesion volume and improved behavioral outcomes. Nanoparticle-enabled delivery of MP presents an effective method for introducing MP locally after SCI and significantly enhances therapeutic effectiveness compared to bare MP administered either systemically or locally. PMID- 19185914 TI - The influence of the stable expression of BMP2 in fibrin clots on the remodelling and repair of osteochondral defects. AB - Growth factors like BMP2 have been tested for osteochondral repair, but transfer methods used until now were insufficient. Therefore, the aim of this study was to analyse if stable BMP2 expression after retroviral vector (Bullet) transduction is able to regenerate osteochondral defects in rabbits. Fibrin clots colonized by control or BMP2-transduced chondrocytes were generated for in vitro experiments and implantation into standardized corresponding osteochondral defects (n=32) in the rabbit trochlea. After 4 and 12 weeks repair tissue was analysed by histology (HE, alcian-blue, toluidine-blue), immunohistochemistry (Col1, Col2, aggrecan, aggrecan-link protein), ELISA (BMP2), and quantitative RT-PCR (BMP2, Col1, Col2, Col10, Cbfa1, Sox9). In vitro clots were also analysed by BMP2-ELISA, histology (alcian-blue), quantitative RT-PCR and in addition by electron microscopy. BMP2 increased Col2 expression, proteoglycan production and cell size in vitro. BMP2 transduction by Bullet was efficient and gene expression was stable in vivo over at least 12 weeks. Proteoglycan content and ICRS-score of repair tissue were improved by BMP2 after 4 and 12 weeks and Col2 expression after 4 weeks compared to controls. However, in spite of stable BMP2 expression, a complete repair of osteochondral defects could not be demonstrated. Therefore, BMP2 is not suitable to regenerate osteochondral lesions completely. PMID- 19185915 TI - Vasculogenesis and angiogenesis in the first trimester human placenta: an innovative 3D study using an immersive Virtual Reality system. AB - First trimester human villous vascularization is mainly studied by conventional two-dimensional (2D) microscopy. With this (2D) technique it is not possible to observe the spatial arrangement of the haemangioblastic cords and vessels, transition of cords into vessels and the transition of vasculogenesis to angiogenesis. The Confocal Laser Scanning Microscopy (CLSM) allows for a three dimensional (3D) reconstruction of images of early pregnancy villous vascularization. These 3D reconstructions, however, are normally analyzed on a 2D medium, lacking depth perception. We performed a descriptive morphologic study, using an immersive Virtual Reality system to utilize the full third dimension completely. This innovative 3D technique visualizes 3D datasets as enlarged 3D holograms and provided detailed insight in the spatial arrangement of first trimester villous vascularization, the beginning of lumen formation within various junctions of haemangioblastic cords between 5 and 7 weeks gestational age and in the gradual transition of vasculogenesis to angiogenesis. This innovative immersive Virtual Reality system enables new perspectives for vascular research and will be implemented for future investigation. PMID- 19185916 TI - Stress differentially modulates mRNA expression for corticotrophin-releasing hormone receptors in hypothalamus, hippocampus and pituitary of prairie voles. AB - This study compares the effect of an acute stressor (restraint for 1h) versus a chronic stressor (social isolation for 4 weeks) on the expression of mRNAs for corticotropin-releasing hormone (CRH), CRH receptor type 1 (CRH-R1) and type 2 (CRH-R2) in the hypothalamus, hippocampus and pituitary of socially monogamous female prairie voles (Microtus ochrogaster). Animals were studied immediately following a stressor or as a function of repairing with a familiar sibling. Despite elevated expression of CRH mRNA, no alteration of CRH-R1 mRNA in the hypothalamus was observed following restraint stress or 4 weeks of social isolation. Hypothalamic CRH-R2 mRNA was significantly lower in voles exposed to restraint or isolation. CRH-R2 mRNA also remained down-regulated in isolated animals when these animals were re-paired with their sibling for one day following 28 days of isolation. Restraint, but not isolation, significantly increased CRH-R1 mRNA and decreased CRH-R2 mRNA in the pituitary. However, these differences were no longer observed when these animals were re-paired with their sibling for one day. Despite elevated CRH mRNA expression, CRH-R1 mRNA did not increase in the hippocampus following restraint or social isolation. Social isolation, but not restraint stress, increased CRH-R2 mRNA in the hippocampus, when these animals were re-paired with their sibling for one day the modulation of CRH mRNA remained up-regulated. Plasma corticosterone was elevated only following restraint, and not in animals that were handled, isolated or re-paired. The results of the present study reveal that acute restraint as well as social isolation can have significant consequences for the modulation of gene expression for the CRH receptors in brain and pituitary of prairie voles. PMID- 19185917 TI - Impact on survival of different treatments for myelodysplastic syndromes (MDS). AB - Therapies for myelodysplastic syndromes (MDS) often achieve hematological responses but their impact on overall survival has generally not been evaluated. The Duesseldorf MDS Registry allowed us to perform matched-pair analyses to assess a possible survival benefit of treatment with thalidomide, valproic acid, low-dose Ara-C, antithymocyte globulin (ATG), induction chemotherapy, or allogeneic stem cell transplantation (allo-SCT). For all treatment modalities, lengthening of survival was restricted to certain subgroups of patients. With the exception of allo-SCT, MDS treatment was generally palliative. Recently, epigenetic treatment with demethylating agents proved to be the first therapy that can significantly prolong survival in patients with higher-risk MDS. PMID- 19185918 TI - Level of DNA topoisomerase IIalpha mRNA predicts the treatment response of relapsed acute leukemic patients. AB - The DNA topoisomerase IIalpha (Topo IIalpha) is known as a target enzyme for many chemotherapeutic agents. We investigated the Topo IIalpha mRNA expression by real time RT-PCR in 37 paired samples at diagnosis and at relapse of acute leukemic patients in relation to drug sensitivity and clinical outcome. The Topo IIalpha levels in leukemic blasts at relapse were significantly higher than that at diagnosis, especially in ALL. The increase in the Topo IIalpha level at relapse was significant in cases which could not achieve a second remission, but not significant in cases which achieved a second remission. These results suggest that the change of Topo IIalpha expression in leukemic blasts at relapse may predict therapeutic responsiveness. PMID- 19185919 TI - Change in the quantity and acute toxicity of pesticides sold in South African crop sectors, 1994-1999. AB - BACKGROUND: South African pesticide market sales data, for two years, 1994 and 1999, were audited to identify change in total and per hectare mass sold and acute toxicity indicator (ATI) (kg sold/rat oral LD50) in the grape, pome, stone fruit, potato and wheat sectors. RESULTS: Total pesticide sales (62%), amount per hectare (42%) and number of active ingredients (23%) increased in 1999 compared to 1994 with the grape fruit sector, the most significant contributor over the two years. Total (14%) and per hectare ATI (19%) decreased in 1999, but not substantially with the potato sector the most significant contributor. CONCLUSIONS: Toxic pesticides were still used in 1999 which highlights a need to develop alternative agricultural and non-chemical pest control methods that reduce usage of pesticides. PMID- 19185920 TI - Flame retardants in indoor dust and air of a hotel in Japan. AB - Occurrence of flame retardants (FRs) in the indoor environment of highly flame retarded public facilities is an important concern from the viewpoint of exposure because it is likely that FRs are used to a greater degree in these facilities than in homes. For this study, brominated flame-retardants (BFRs) and organophosphate flame-retardants and plasticizers (OPs), and brominated dibenzo-p dioxins/furans (PBDD/DFs) were measured in eight floor dust samples taken from a Japanese commercial hotel that was assumed to have many flame-retardant materials. Concentrations of polybrominated diphenylethers (PBDEs) and hexabromocyclododecanes (HBCDs) varied by about two orders of magnitude, from 9.8 1700 ng/g (median of 1200 ng/g) and from 72-1300 ng/g (median of 740 ng/g), respectively. Concentrations of the two types of BFRs described above were most dominant among the investigated BFRs in the dust samples. It is inferred that BFR and PBDD/DF concentrations are on the same level as those in house and office dust samples reported based on past studies. Regarding concentrations of 11 OPs, 7 OPs were detected on the order of micrograms per gram, which are equivalent to or exceed the BFR concentrations such as PBDEs and HBCDs. Concentrations of the investigated compounds were not uniform among dust samples collected throughout the hotel: concentrations differed among floors, suggesting that localization of source products is associated with FR concentrations in dust. Passive air sampling was also conducted to monitor BFRs in the indoor air of hotel rooms: the performance of an air cleaner placed in the room was evaluated in terms of reducing airborne BFR concentrations. Monitoring results suggest that operation of an appropriate air cleaner can reduce both gaseous and particulate BFRs in indoor air. PMID- 19185922 TI - Reaching and grasping a moving object in 6-, 8-, and 10-month-old infants: laterality and performance. AB - The goal of this study was to investigate some of the visuo-motor factors underlying an infant's developing ability to grasp a laterally-moving object. In particular, hand preference, midline crossing, and visual-field asymmetry were investigated by comparing performance as a function of the object's direction of motion. We presented 6-, 8-, and 10-month-old infants with a graspable object, moving in a circular trajectory in the horizontal plane. Six-month-old infants reached for the object with the ipsilateral hand and grasped it with the contralateral hand. Eight-month-old infants showed a strong right-hand bias for both reaching and grasping. Ten-month-old infants showed a greater diversity of strategy use including bimanual and successful ipsilateral grasping following ipsilateral reaching in both directions of motion. Thus, motor constraints due to spatial compatibility, hand preference and bimanual coordination (but not midline crossing) must be taken into account to understand age differences in grasping a moving object. PMID- 19185921 TI - Functional CD1d and/or NKT cell invariant chain transcript in horse, pig, African elephant and guinea pig, but not in ruminants. AB - CD1d-restricted invariant natural killer T cells (NKT cells) have been well characterized in humans and mice, but it is unknown whether they are present in other species. Here we describe the invariant TCR alpha chain and the full length CD1d transcript of pig and horse. Molecular modeling predicts that porcine (po) invariant TCR alpha chain/poCD1d/alpha-GalCer and equine (eq) invariant TCR alpha chain/eqCD1d/alpha-GalCer form complexes that are highly homologous to the human complex. Since a prerequisite for the presence of NKT cells is the expression of CD1d protein, we performed searches for CD1D genes and CD1d transcripts in multiple species. Previously, cattle and guinea pig have been suggested to lack CD1D genes. The CD1D genes of European taurine cattle (Bos taurus) are known to be pseudogenes because of disrupting mutations in the start codon and in the donor splice site of the first intron. Here we show that the same mutations are found in six other ruminants: African buffalo, sheep, bushbuck, bongo, N'Dama cattle, and roe deer. In contrast, intact CD1d transcripts were found in guinea pig, African elephant, horse, rabbit, and pig. Despite the discovery of a highly homologous NKT/CD1d system in pig and horse, our data suggest that functional CD1D and CD1d-restricted NKT cells are not universally present in mammals. PMID- 19185923 TI - Susceptibility of the C2 canine mastocytoma cell line to the effects of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a member of the TNF family, which preferentially induces apoptosis in cells that have undergone malignant transformation. In humans, non-neoplastic cells are normally protected from the effects of TRAIL by expressing decoy receptors, lacking death domains. In contrast, neoplastic cells tend to downregulate their decoy receptor expression, increasing their susceptibility to the pro-apoptotic effects of TRAIL, via the functional TRAIL receptors. The aim of the current study was to investigate the effect of TRAIL on the canine C2 mastocytoma cell line to determine whether this agent might be a suitable treatment for mast cell tumors in dogs. C2 and MDCK cells were cultured with recombinant human TRAIL. Apoptosis was assessed using a Caspase 3 and 7 chemiluminescence assay and flow cytometry following Annexin V:FITC labelling. Cell metabolism was assessed using a colorimetric MTT-based assay. C2 cells demonstrated greater sensitivity to TRAIL induced apoptosis compared to MDCK cells by all assessment methods. The dog genome assembly was searched for orthologs of TRAIL and its receptors using published sequences from other species for reference. Although a canine ortholog for TRAIL was identified, only one TRAIL receptor ortholog (TNFRSF11B) could be found. C2, but not MDCK, cells expressed mRNA for TNFRSF11B, detected by RT-PCR. In other species, TNFRSF11B is a decoy receptor, as even though it has a death domain it is secreted due to its lack of a transmembrane domain. The effect of TRAIL on the C2 cell line suggests that this cytokine might be suitable for treatment of mast cell tumors in dogs. PMID- 19185924 TI - Newborn hearing screening and strategy for early detection of hearing loss in infants. AB - OBJECTIVE: More than 80% of permanent hearing losses (HL) in children are congenital. Newborn hearing screening (NHS) is the best method for early detection of suspected hearing loss. If the NHS is not universal more than 30% permanent hearing losses are not identified. There are various methods of NHS: otoacoustic emissions (TEOAE, DPOAE) and automatic auditory brainstem response (AABR). After hearing screening, and when hearing loss is suspected, tympanometry and audiological methods then used for determination of hearing threshold; these include ABR, ASSR or/and behavioral methods. The goal of this study is to evaluate the influence of UNHS on the early detection of hearing loss in children before and after the implementation of obligatory universal newborn hearing screening in Slovakia, and also on the etiologic evaluation of hearing impaired infants identified by screening. METHOD: In Slovakia NHS started in 1998 and was provided in ENT departments. From May 1, 2006 UNHS has been mandatory in Slovakia, using two stages TEOAE in all newborn departments in Slovakia (64 newborn departments). In year 2005--42% of newborns in Slovakia were screened, in 2006--66% newborns and in 2007--94, 99% (three small newborn departments do not yet have equipment for OAE screening). For determination of hearing thresholds ASSR are used in two ENT departments and ABR in the other four ENT departments. RESULTS: Comparing the number of identified cases with bilateral severe permanent HL or deafness before and after UNHS, 22.8% more cases of PHL were identified in the first year of UNHS. Also the average age of diagnosis of PHL was lower. In the year 2007, 94% of newborns were screened. We found 0.947/1000 newborns with bilateral severe PHL (35.9%) more than before UNHS). After audiologic and etiologic assessment of the 76 infants who failed screening, 5 (6.58%) were found to have normal hearing, 16 (22.54%) had unilateral and 55 (77.46%) had bilateral SNHL. A non-syndromic genetic cause was present in 25.45% of cases, syndromic in 9%, perinatal cause (31%), congenital CMV infection in 7.27%, bilateral cochlear anomalies without other abnormality in 1.83% and unknown etiology in 25.45%. PMID- 19185925 TI - IJPORL and ESPO: congratulations and a short history. PMID- 19185926 TI - A comparison of a child's fundamental frequencies in structured elicited vocalizations versus unstructured natural vocalizations: a case study. AB - OBJECTIVES: Building on the concept that task type may influence fundamental frequency (F(0)) values, the purpose of this case study was to investigate the difference in a child's F(0) during structured, elicited tasks and long-term, unstructured activities. It also explores the possibility that the distribution in children's F(0) may make the standard statistical measures of mean and standard deviation less than ideal metrics. METHODS: A healthy male child (5 years, 7 months) was evaluated. The child completed four voice tasks used in a previous study of the influence of task type on F(0) values: (1) sustaining the vowel /a/ in isolation; (2) sustaining the vowel /a/ embedded in a word at the end of a phrase; (3) repeating a sentence; and (4) counting from 1 to 10. The child also wore a National Center for Voice and Speech voice dosimeter, a device that collects voice data over the course of an entire day, during all activities for 34 h over 4 days. RESULTS: Throughout the structured vocal tasks within the clinical environment, the child's F(0), as measured by both the dosimeter and acoustic analysis of microphone data, was similar for all four tasks, with the counting task the most dissimilar. The mean F(0) (approximately 257 Hz) matched very closely to the average task results in the literature given for the child's age group. However, the child's mean fundamental frequency during the unstructured activities was significantly higher (approximately 376 Hz). Finally, the mode and median of the structured vocal tasks were 260 Hz and 259 Hz respectively (both near the mean), while the unstructured mode and median were 290 Hz and 355 Hz respectively. CONCLUSIONS: The results of this study suggest that children may produce a notably different voice pattern during clinical observations compared to routine daily activities. In addition, the child's long term F(0) distribution is not normal. If this distribution is consistent in long term, unstructured natural vocalization patterns of children, statistical mean would not be a valid measure. Mode and median are suggested as two parameters which convey more accurate information about typical F(0) usage. Finally, future research avenues, including further exploration of how children may adapt their F(0) to various environments, conversation partners, and activity, are suggested. PMID- 19185927 TI - Preliminary study of proinflammatory cytokines and chemokines in the middle ear of acute otitis media due to Alloiococcus otitidis. AB - Alloiococcus otitidis is a newly discovered organism frequently detected in otitis media. However, the association of the organism with the development of otitis media has not been disclosed in detail yet. In the middle ear, proinflammatory cytokines and chemokines are released in association with infection by pathogens, and these cytokines contribute to the induction of an inflammatory reaction. To investigate the profile of inflammation-related cytokines in the acute phase of A. otitidis infection, we analyzed the release of proinflammatory cytokines and chemokines in middle ear effusions of acute otitis media due to A. otitidis, in comparison with acute otitis media due to the well known Gram-positive middle ear pathogen Streptococcus pneumoniae. The amounts of proinflammatory cytokines (IL-8, IL-1beta, IL-6, TNF-alpha) and CXC chemokines (IP-10, I-TAC) were significantly increased in the A. otitidis group as well as in the S. pneumoniae group. Various inflammation-related cytokines/chemokines were induced in the A. otitidis-infected middle ear, and the profile of cytokines was very similar to that in S. pneumoniae infection. This preliminary study suggests that A. otitidis has the potential to induce these cytokines, contributing to the development of an inflammatory reaction in the middle ear cavity in a similar manner to S. pneumoniae. PMID- 19185928 TI - Effects of naphthalene, beta-naphthoflavone and benzo(a)pyrene on the diurnal and nocturnal indoleamine metabolism and melatonin content in the pineal organ of rainbow trout, Oncorhynchus mykiss. AB - Polycyclic aromatic hydrocarbons (PAHs) have deleterious effects on neuroendocrine systems in teleost fish affecting, among other processes, reproductive function or stress responses. The hormone melatonin, mainly produced in the pineal organ of vertebrates, is involved in the regulation of biological rhythms as well as other important functions, and may also act as an antioxidant molecule. The effects of environmental pollutants on the endocrine and metabolic activity of the pineal organ have been studied only in mammals. We here evaluate the effects of the PAHs naphthalene (NAP) and benzo(a)pyrene (BaP) and the flavonoid beta-naphthoflavone (BNF) on the pineal organ of rainbow trout by quantifying the diurnal and nocturnal pineal content of some indoles and methoxyindoles, including melatonin. NAP mainly induced diurnal increases in the pineal content of melatonin and other methoxyindoles like 5-methoxytryptamine (5 MT), 5-methoxyindole-3-acetic acid (5-MIAA) or 5-methoxytryptophol (5-MTOL). Those increases did not occur at night, when even occasional decreases were observed compared with controls. NAP also induced some diurnal and nocturnal decreases in the levels of indolic compounds like serotonin (5-HT) and 5 hydroxyindole-3-acetic acid (5-HIAA), while pineal content of 5-hydroxytryptophan (5-HTP) was first decreased (few hours after injection) and then increased (few days after injection) during the day. BaP and BNF induced strong increases in diurnal levels of melatonin, whereas other pineal compounds were unaffected. It seems that an increase of the methylation capacity of the pineal organ takes place during the day, and a decrease occurs at night. Those effects could be mediated by changes in the activity of key enzymes involved in pineal melatonin biosynthesis, maybe as a result of the alteration of the cellular phototransduction mechanisms involved in the light-induced inhibition of melatonin synthesis in the pineal photoreceptor cells. These results demonstrate for the first time that environmental pollutants can disrupt the activity of the pineal organ of teleost fish. This disruption could be a threat for the survival of the animals in their natural environment, although the increases observed in melatonin levels could play a relevant role as a toxicity-protection factor. PMID- 19185929 TI - Massive melanotic myocardial metastasis characterized by multiple cardiac imaging modalities. AB - Melanoma has a great propensity for cardiac involvement but most cases are diagnosed post-mortem. We present the case of a 67-year-old man with history of malignant melanoma who presented with increasing dyspnea, fatigue and peripheral edema. A large myocardial metastasis was identified on transthoracic echocardiography, and further characterized with cardiovascular magnetic resonance prior to surgery. PMID- 19185930 TI - Myocarditis and cardiomyopathy HIV associated. AB - Heart muscle involvement associated with human immunodeficiency virus (HIV) infection may present as myocarditis, dilated cardiomyopathy or as isolated left or right ventricular dysfunction. Histopathological and ultra structural findings with different degrees of cardiac-chamber dilation have been described and an important role of the cytokines tumor necrosis factor-alpha (TNF-alpha), interleukin-1 (IL-1) and IL-6 has been suggested. We present a case of myocarditis in a 47-year-old woman with HIV associated cardiomyopathy, focussing attention on heart muscle involvement in HIV disease. PMID- 19185931 TI - Prostate-specific antigen kallikrein and acute myocardial infarction: where we are. Where are we going? AB - Prostate-specific antigen (PSA) is an established marker for the detection of prostate cancer. Both elevated and diminished PSA have been reported during acute myocardial infarction. It seems that when elevation of PSA occurs during acute myocardial infarction (AMI), coronary lesions are frequent and often more severe than when a diminution of PSA occurs. PSA has been identified as a member of the human kallikrein family of serine proteases. In recent years, numerous observations have suggested that the activity of the kallikrein-kinin system is related to inflammation and to cardiovascular diseases. PSA kallikrein, however, does not seem to have kinin-generating activity. The inactive precursor form of PSA, proPSA, is converted rapidly to active PSA by Human kallikrein 2 (hK2), suggesting an important in vivo regulatory function byhK2 on PSA activity. However, it has been reported that hK2 might not alone be able to activate proPSA in vivo, but there are also other protease/proteases involved in this event. Moreover, it seems that when elevation of prostate-specific antigen occurs during AMI, it seems to relate to a higher occurrence of major adverse cardiac events in the first 8 days after AMI than when a diminution of PSA occurs. It confirms a possible new intriguing scenario of the role of the PSA in AMI. Although these preliminary observations are suggestive, large studies need to be done to confirm these preliminary results. PMID- 19185932 TI - Atrial fibrillation associated with exogenous subclinical hyperthyroidism. AB - Subclinical hyperthyroidism is an increasingly recognized entity that is defined as a normal serum free thyroxine and free triiodothyronine levels with a thyroid stimulating hormone level suppressed below the normal range and usually undetectable. It has been reported that subclinical hyperthyroidism is not associated with coronary heart disease or mortality from cardiovascular causes but it is sufficient to induce arrhythmias including atrial fibrillation and atrial flutter. It has also been reported that increased factor X activity in patients with subclinical hyperthyroidism represents a potential hypercoagulable state. Moreover acute myocardial infarction has been reported during L-thyroxine substitution therapy. Far more common and relatively less studied is exogenous subclinical hyperthyroidism caused by L-thyroxine administration to thyroidectomized or hypothyroid patients or patients with simple or nodular goiter. We present a case of atrial fibrillation associated with exogenous subclinical hyperthyroidism, in a 72-year-old Italian woman. Also this case focuses attention on the importance of a correct evaluation of subclinical hyperthyroidism. PMID- 19185933 TI - Management of acute coronary syndromes in developing countries: are we complying with practice guidelines? PMID- 19185934 TI - Clinical management and short-term cost - 64-slice MDCT vs. myocardial perfusion scintigraphy. AB - There are currently no published studies of the impact of CT coronary angiography (CTA) on patient management or cost when compared with other established imaging techniques. We assessed the short term investigation and treatment costs of CTA compared with myocardial perfusion scintigraphy (MPS) using real clinical scenarios. Clinical information with either their CTA or MPS results were presented to 20 cardiologists in a random order. They decided further investigations and treatment required based on these data. Short term cost was calculated for each imaging strategy. Whilst the total number of further investigations requested did not differ between groups patients undergoing CTA were more likely to be referred for invasive coronary angiography, receive aspirin, statins, ACE inhibitors, beta-blockers or clopidogrel. Overall cost and investigation costs were similar between CTA and MPS; however treatment costs were higher with CTA. There are significant differences in further clinical management when using CTA compared with MPS, in particular with greater use of secondary preventative medication. PMID- 19185935 TI - Left ventricular thrombus complicating tachycardia-induced cardiomyopathy. AB - A 45 day old new-born with arrhythmia-induced cardiomyopathy complicated by thrombus formation is presented. Drug treatment produced immediate symptomatic relief and subsequent reversion to normal cardiac function. The thrombus disappeared a few days later. PMID- 19185936 TI - Multi-detector computerized tomography angiography for evaluation of acute chest pain--a meta analysis and systematic review of literature. AB - BACKGROUND: Evaluation of pain localized to the chest in the emergency room is, challenging, time-consuming, costly, and often inconclusive. Available research, though limited, suggests a role for MDCTA in the evaluation of patients with acute chest pain of low to intermediate risk, for identifying and excluding ACSs during the initial emergency department evaluation. Accordingly, our aim was to conduct a meta-analysis to assess the diagnostic accuracy of MDCTA in this setting. METHODS: We included all studies that compared MDCTA with either coronary angiography or standard of care for early and accurate triage of patients presenting with acute chest pain. Published studies were identified by searches of the Pubmed, Ovid and Google scholar databases as well as hand searches of selected references. Data were extracted independently by two reviewers. Included studies were evaluated for heterogeneity. Meta-analysis was performed at patient level using a random-effects model. RESULTS: 16 studies totaling 1119 patients were included in the current meta-analysis: one randomized trial, one retrospective analysis and fourteen prospective cohort studies. Pooled DOR was 190.80 (95%CI, 102.94-353.65). The pooled sensitivity and specificity were 0.96 (95%CI, 0.93-0.98) and 0.92(95%CI, 0.89-0.94) respectively. The pooled NLR and PLR were 0.09 (95%CI, 0.06-0.14) and 10.12 (95%CI, 6.73-15.22). CONCLUSION: MDCTA has an excellent diagnostic accuracy in detection of significant coronary artery stenosis in patients with acute chest pain. This diagnostic accuracy of MDCTA has a potential for rapid triage of patients in the ED, with acute chest pain of low to intermediate risk of acute coronary syndrome, to rule out significant epicardial stenosis as the etiology of chest pain. PMID- 19185937 TI - A rare case of reversible constrictive pericarditis with severe pericardial thickening in a patient with adult onset Still's disease. AB - A 21-year-old male German patient, was referred to our intensive care unit from a peripheral hospital with severe septic illness and fever. The echocardiographic investigation revealed a massive pericardial thickening of at least 20 mm mainly located posterolateral and inferior, without typical pericardial effusion. An inspiratory diastolic septal shift to the left, as well as pathological respiratory PW-Doppler profiles of mitral and tricuspid inflow velocities could clearly been detected in the examination. The patient describes at the interview in our intensive care unit, that he has previously been well, without any symptoms of cardiac failure or joint pain. We diagnosed an adult onset Still's disease (AOSD). Thus, prednisone pulse-therapy was initiated with 250 mg and reduced stepwise in the following days. All the symptoms improved dramatically within a few days. The documented pericardial thickening showed a significant reduction to approximately 6 mm already after 4 days of therapy AOSD should not be forgotten in the differential diagnosis of pericardial disease due to its potentially life threatening complications. Usually, echocardiography reveals typical pericardial effusion. In this case the patient presented with an impressing, atypical thickening of the pericardium in the posterolateral region, hardly ever seen in AOSD. PMID- 19185938 TI - A congenital form of junctional ectopic tachycardia. AB - Accessory pathways have been described as well as their Ecg identification criteria also in pediatric population. Radiofrequency ablation is a curative treatment but its application has been more limited in the paediatric population. The congenital form of junctional ectopic tachycardia was firstly described by Coumel et al. in 1976. It usually occurs in the first six months of life presenting as a persistent sustained form, lasting up to 90% of the time and it is hampered by high mortality. Its clinical presentation may be dramatic, being associated in up to 60% of cases with cardiomegaly and/or heart failure. Secondary dilated cardiomyopathy, ventricular fibrillation and sudden cardiac death have also been reported. We present a case of congenital form of junctional ectopic tachycardia in a 12-day-old newborn infant. Also this case is illustrative of the congenital form of junctional ectopic tachycardia. PMID- 19185939 TI - Atrial fibrillation and acute myocardial infarction without significant coronary stenoses associated with subclinical hyperthyroidism and erythrocytosis. AB - Subclinical hyperthyroidism is an increasingly recognized entity that is defined as a normal serum free thyroxine and free triiodothyronine levels with a thyroid stimulating hormone level suppressed below the normal range and usually undetectable. It has been reported that sub-clinical hyperthyroidism is not associated with CHD or mortality from cardiovascular causes but is sufficient to induce arrhythmias including atrial fibrillation and atrial flutter. Moreover increased factor X activity in patients with subclinical hyperthyroidism represents a potential hypercoagulable state. It has been also reported an acute myocardial infarction with normal coronary arteries associated with iatrogenic hyperthyroidism and with a myocardial bridge too. It has been also reported an acute myocardial infarction without significant coronary stenoses associated with subclinical hyperthyroidism. Furthermore it has been reported that at highly increased hematocrit levels patients may experience hyperviscosity symptoms. We present a case of atrial fibrillation and acute myocardial infarction without significant coronary stenoses associated with subclinical hyperthyroidism and erythrocytosis. Also this case focuses attention on the importance of a correct evaluation of subclinical hyperthyroidism. PMID- 19185940 TI - Atheroscerlotic heart disease in Bangladeshi immigrants: risk factors and angiographic findings. AB - BACKGROUND: The prevalence of coronary artery disease (CAD) among Bangladeshis greatly exceeds that of Caucasians. Bangladeshis also suffer from premature onset, clinically aggressive and angiographically extensive disease. The role of conventional CAD risk factors (CCRFs) has been questioned. We therefore sought to determine if the CCRFs of Bangladeshis differed from non-Bangladeshis. We also sought to determine whether CAD was more extensive in Bangladeshis and if Bangladeshi ethnicity was independently predictive of extensive i.e., 3-vessel CAD at angiography. METHODS: We reviewed the coronary angiograms and medical records of 75 Bangladeshis and 57 non-Bangladeshis presenting with myocardial infarction or angina pectoris. RESULTS: Bangladeshis were younger (56.1 vs. 62.4 years, p=.001), had a lower body-mass index (25.2 vs. 27.2 kg/m(2), p=.017) and were less likely to be current or recent smokers (40% vs. 58%, p=.041) than non Bangladeshis. There were no statistically significant differences in the proportion of subjects in the 2 groups with respect to diabetes mellitus, dyslipidemia, hypertension or family history of CAD. Bangladeshis had twice the rate of 3-vessel CAD of non-Bangladeshis (53% vs. 26%, p=.002). Bangladeshi ethnicity was independently associated with >3X the likelihood of having 3-vessel CAD at angiography (p=.011). CONCLUSIONS: This study demonstrated that the CCRF burden of Bangladeshis with CAD is not excessive compared to that of non Bangladeshis and is therefore unlikely to account for the excessive CAD risk found in this cohort. We also conclude that Bangladeshis have more angiographically extensive CAD than non-Bangladeshis and that Bangladeshi ethnicity is independently predictive of 3-vessel disease. PMID- 19185941 TI - Irrigation linked to a greater incidence of human and veterinary West Nile virus cases in the United States from 2004 to 2006. AB - Identifying risk factors for West Nile virus transmission is vital for future public health control measures. This study investigated the effect of irrigation, expressed as percent irrigated land, and population density on the incidence of human and veterinary West Nile virus (WNV) cases using a sample of 2827 counties from the United States over a 3-year period. Zero-inflated Poisson regression models were selected because of the large number of counties reporting zero cases. The mean percentage of irrigated land for the entire sample was approximately 0.029, while counties reporting both human and veterinary cases had an average of 0.054. As the level of irrigation rose by 0.1% of the total county land area, the incidence rate of WNV cases increased by 50% in the human model (RR: 1.50, 95% CI: 1.46-1.53) and 63% in the veterinary model (RR: 1.63, 95% CI: 1.57-1.68). Higher equine and human population densities were associated with a slightly decreased incidence of WNV cases. In the logistic portion of the Zero inflated model, the presence of irrigation significantly decreased the odds of a county reporting zero cases. Future research is warranted to determine the exact nature of the relationship between irrigation and increased disease incidence in human and veterinary populations. PMID- 19185942 TI - The effect of income growth and inequality on health inequality: Theory and empirical evidence from the European Panel. AB - Governments of EU countries have declared that they would like to couple income growth with reductions in social inequalities in income and health. We show that, theoretically, both aims can be reconciled only under very specific conditions concerning the type of growth and the income responsiveness of health. We investigate whether these conditions were met in Europe in the 1990s using panel data from the European Community Household Panel. We demonstrate that (i) in most countries, the income elasticity of health was positive and increases with income, and (ii) that income growth was not pro-rich in most EU countries, resulting in small or negligible reductions in income inequality. The combination of both findings explains the modest increases we observe in income-related health inequality in the majority of countries. PMID- 19185943 TI - Knockout of three-component regulatory systems reveals that the apparently constitutive plantaricin-production phenotype shown by Lactobacillus plantarum on solid medium is regulated via quorum sensing. AB - It has been found that many bacteriocins from lactic acid bacteria (LAB) are only produced in broth cultures when specific growth conditions are achieved and a dedicated three-component regulatory system, involved in a quorum sensing (QS) mechanism, is switched on. Surprisingly, bacteriocin production in LAB occurs in an apparently constitutive manner on solid media. This study addresses the question of constitutive versus regulated bacteriocin production on solid media in two different QS-regulated plantaricin-producing strains: Lactobacillus plantarum NC8 and L. plantarum WCFS1. Construction of knockout mutants for their respective regulatory operons revealed that bacteriocin production is controlled through a QS mechanism in both strains, on solid as well as in liquid media. These results could be extensible to other bacteriocins from LAB which are only produced on agar plates and not in broth cultures. Our findings suggest that QS regulated bacteriocin production in LAB has evolved for competing on solid supports rather than in liquid media. In practice, this could be of major importance in vegetable fermentations, where the solid substrate itself provides an enormous surface where bacteria can attach to and produce biofilms. Therefore, QS-regulated bacteriocinogenic LAB growing in biofilms are under the optimum conditions to produce bacteriocins. Selection of strains to be used as starter cultures for vegetable fermentations should take into account these facts. PMID- 19185944 TI - Visfatin in gestational diabetes: serum level and mRNA expression in fat and placental tissue. AB - AIMS AND METHODS: In this study we measured: (1) serum visfatin concentrations in pregnant women with normal glucose tolerance (NGT) and gestational diabetes mellitus (GDM) between 26 and 33 weeks of gestation, using two immunoassays: EIA and ELISA; (2) serum visfatin levels (ELISA) and its mRNA expression (quantitative real-time PCR) in subcutaneous adipose tissue (SAT), visceral adipose tissue (VAT) and placental tissue from women with NGT and GDM at term. RESULTS: Visfatin concentrations (measured by EIA and ELISA) did not differ in the women with GDM and NGT between 26 and 33 weeks of gestation but were significantly lower in GDM than in NGT subjects at term (2.7 [0.7-4.6] vs 5.2 [3.7-5.9]ng/ml, p=0.02). There was no difference in visfatin mRNA expression in fat and placental tissue between the two subgroups. Regression analysis revealed that visfatin mRNA expression was significantly related to interleukin-6 and tumour necrosis factor-alpha mRNA expression in SAT (beta=0.39, p=0.009 and beta=0.47, p=0.002) and placental tissue (beta=0.37, p=0.03 and beta=0.49, p=0.005). CONCLUSIONS: Circulating visfatin was significantly lower in the GDM than in the NGT subjects at term, although no differences in its mRNA expression in fat and placental tissues were observed. PMID- 19185945 TI - Does subsidy work? Price elasticity of demand for influenza vaccination among the elderly in Japan. AB - OBJECTIVES: Subsidy for influenza vaccination is often provided to the elderly in order to encourage them to receive a flu shot in developed countries. However, its effect on uptake rate, i.e., price elasticity of demand, has not been well studied. METHODS: Japan's decentralised vaccination programme allows observation of various pairs in price and uptake rate of flu shots among the elderly by the municipality from 2001/2002 to 2004/2005 season. We combine our sample survey data (n=281), which monitor price, subsidy and uptake rate, with published data on local characteristics in order to estimate price elasticity of demand with panel model. RESULTS: We find price elasticity of demand for influenza vaccine: nearly zero in nationwide, nearly zero in urban area, and -1.07 in rural area. CONCLUSIONS: The results question the rationale for subsidy, especially in urban area. There are cases where maintaining or increasing the level of subsidy is not an efficient allocation of finite health care resources. When organising a vaccination programme, health manager should be careful about the balance between subsidy and other efforts in order to encourage the elderly to receive shots with price elasticity in mind. PMID- 19185946 TI - Arrowsmith two-node search interface: a tutorial on finding meaningful links between two disparate sets of articles in MEDLINE. AB - The Arrowsmith two-node search is a strategy that is designed to assist biomedical investigators in formulating and assessing scientific hypotheses. More generally, it allows users to identify biologically meaningful links between any two sets of articles A and C in PubMed, even when these share no articles or authors in common and represent disparate topics or disciplines. The key idea is to relate the two sets of articles via title words and phrases (B-terms) that they share. We have created a free, public web-based version of the two-node search tool (http://arrowsmith.psych.uic.edu), have described its development and implementation, and have presented analyses of individual two-node searches. In this paper, we provide an updated tutorial intended for end-users, that covers the use of the tool for a variety of potential scientific use case scenarios. For example, one can assess a recent experimental, clinical or epidemiologic finding that connects two disparate fields of inquiry--identifying likely mechanisms to explain the finding, and choosing promising follow-up lines of investigation. Alternatively, one can assess whether the existing scientific literature lends indirect support to a hypothesis posed by the user that has not yet been investigated. One can also employ two-node searches to search for novel hypotheses. Arrowsmith provides a service that cannot be carried out feasibly via standard PubMed searches or by other available text mining tools. PMID- 19185947 TI - Estimating the measuring sensitivity of unipolar and bipolar ECG with lead field method and FDM models. AB - New portable electrocardiogram (ECG) measurement systems are emerging into market. Some use nonstandard bipolar electrode montage and sometimes very small interelectrode distances to improve the usability of the system. Modeling could provide a straightforward method to test new electrode systems. The aim of this study was to assess whether modeling the electrodes' measuring sensitivity with lead field method can provide a simple tool for testing a number of new electrode locations. We evaluated whether the actual ECG signal strength can be estimated by lead fields with two realistic 3D finite difference method (FDM) thorax models. We compared the modeling results to clinical body surface potential map (BSPM) data from 236 normal patients and studied 117 unipolar and 42 bipolar leads. In the case of unipolar electrodes the modeled measuring sensitivities correlated well with the clinical data (r=0.86, N=117, p<0.05). In the case of bipolar electrodes the correlation was moderate (r=0.62 between Model 1 and clinical data, r=0.71 between Model 2 and clinical data, N=42 and p<0.05 for both). Based on this we can conclude that lead field analysis based on realistic thorax models provides a good initial prediction for designing new electrode montages and measurement systems. PMID- 19185949 TI - Comprehensive analysis of EGFR signaling pathways in Japanese patients with non small cell lung cancer. AB - PURPOSE: Translational approach is essentially needed to return the achievement of basic researches to oncological practice. The molecular associations among EGFR mutation and the components of EGFR signaling pathways have been extensively studied in laboratory experiments, although were still controversial. Moreover, the impact of downstream signaling of EGFR on clinical features in patients with non-small cell lung cancer (NSCLC) remains undetermined. PATIENTS AND METHODS: A total of 93 surgically resected NSCLC patients were recruited the study. EGFR mutation status was analyzed by direct sequence method. The protein expression levels of EGFR, phosphorylated EGFR (pEGFR), phosphorylated Akt (pAkt), and phosphorylated MAPK (pMAPK) were determined by immunohistochemistry. RESULTS: There were 37 (40%) patients whose tumor harboring EGFR mutations (1 in exon 18, 22 in exon 19, and 14 in exon 21). Protein expression of EGFR, pEGFR, pAkt, and pMAPK was detected in 61 (66%), 27 (29%), 58 (62%), and 41 (44%) patients, respectively. The expression of pAkt was significantly associated with female gender and never-smoking history, and it was frequently upregulated in tumors harboring EGFR mutations (p<0.05, each). Phosphorylation of EGFR was closely correlated with the EGFR protein expression (p<0.05), but not with the EGFR mutations. In regard to patient survival, none of the molecular biomarkers was predictive for survival after surgical resection, but pMAPK expression was predictive for poor prognosis after gefitinib treatment in patients with postoperative recurrence (p<0.05), suggesting the strong linkage between pMAPK expression and survival benefit from gefitinib. CONCLUSION: Our result could provide new insight into MAP kinase signaling when we treat NSCLC patients with gefitinib. PMID- 19185950 TI - Cigarette smoking and changing trends of lung cancer incidence by histological subtype among Chinese male population. AB - We analyzed the time trends of lung cancer by histological subtype in Hong Kong during 1991-2005, and examined how the time trends were influenced by the effects of birth cohort and calendar period of diagnosis. Cancer incidence data were obtained from Hong Kong Cancer Registry and population data from Census and Statistics Department. Age-standardized incidence rates were computed by the direct method using WHO 1966 standard population as reference. Period and cohort effects were assessed by using two separate Poisson regression models adjusting for age. From 1991 to 2005, the incidence rates in Hong Kong Chinese males decreased steadily. The decline in overall lung cancer incidence rates was limited primarily to the decrease in squamous cell carcinoma, which could be explained by the decreasing trend of cigarette smoking. Adenocarcinoma had been the most predominant histological subtype all along. The relatively horizontal trend of adenocarcinoma and the lack of cohort effect implied the important roles of gene-environment interaction and/or the use of low-tar and filter tip cigarettes. Our study suggests that different histological subtypes may represent different disease entities with perhaps some distinct risk factors. The hypotheses generated from this ecological study will need confirmation by subsequent analytic studies. PMID- 19185948 TI - Dietary chemoprevention strategies for induction of phase II xenobiotic metabolizing enzymes in lung carcinogenesis: A review. AB - Lung cancer is the leading cause of cancer mortality for men and women in the United States and is a growing worldwide problem. Protection against lung cancer is associated with higher dietary intake of fruits and vegetables, according to recent large epidemiologic studies. One strategy for lung cancer chemoprevention focuses on the use of agents to modulate the metabolism and disposition of tobacco, environmental and endogenous carcinogens through upregulation of detoxifying phase II enzymes. We summarize the substantial evidence that suggests that induction of phase II enzymes, particularly the glutathione S-transferases, plays a direct role in chemoprotection against lung carcinogenesis. The engagement of the Keap1-Nrf2 complex regulating the antioxidant response element (ARE) signaling pathway has been identified as a key molecular target of chemopreventive phase II inducers in several systems. Monitoring of phase II enzyme induction has led to identification of novel chemopreventive agents such as the isothiocyanate sulforaphane, and the 1,2-dithiole-3-thiones. However, no agents have yet demonstrated clear benefit in human cell systems, or in clinical trials. Alternative strategies include: (a) using intermediate cancer biomarkers for the endpoint in human trials; (b) high-throughput small molecule discovery approaches for induced expression of human phase II genes; and (c) integrative approaches that consider pharmacogenetics, along with pharmacokinetics and pharmacodynamics in target lung tissue. These approaches may lead to a more effective strategy of tailored chemoprevention efforts using compounds with proven human activity. PMID- 19185951 TI - Ecological explanations for (incomplete) speciation. AB - Divergent natural selection has been shown to promote speciation in many taxa. However, although divergent selection often initiates the process of speciation, it often fails to complete it. Several time-based, geographic and genetic factors have been recognized to explain this variability in how far speciation proceeds. We review here recent evidence indicating that variability in the completeness of speciation can also be associated with the nature of divergent selection itself, with speciation being greatly promoted by (i) stronger selection on a given, single trait (the 'stronger selection' hypothesis) and (ii) selection on a greater number of traits (the 'multifarious selection' hypothesis). However, evidence for each selective hypothesis is still scarce, and further work is required to determine their relative importance. PMID- 19185952 TI - Jasmonate-induced ripening delay is associated with up-regulation of polyamine levels in peach fruit. AB - Methyl jasmonate (MJ, 0.20mM) and its synthetic analog n-propyl dihydrojasmonate (PDJ, 0.22mM) were applied to peach fruit (Prunus persica L. Batsch) at a late developmental stage under field conditions (in planta). On the basis of a previously demonstrated jasmonate (JA)-induced ripening delay in peach, the effects of JAs on the time course of the endogenous polyamine (PA) accumulation and expression of their biosynthetic genes arginine decarboxylase (ADC), ornithine decarboxylase (ODC), spermidine synthase (SPDS) and S adenosylmethionine decarboxylase (SAMDC) were evaluated in control and JA-treated fruit during the 21-d trial period. In parallel, the main ripening-related parameters (ethylene production, flesh firmness and soluble solids contents) were measured, and transcription profiles of aminocyclopropane-1-carboxylic acid oxidase (PpACO1) and of two ethylene perception genes were evaluated. PDJ, but not MJ, reduced ethylene production and fruit softening, impaired PpACO1 transcription and altered the expression of PpERS1 (ethylene sensor 1), but not the expression of PpETR1 (ethylene receptor 1). In the epicarp and mesocarp, the pattern of PA accumulation was altered in a biphasic manner leading to a higher overall PA level in PDJ-treated fruit. Short and long term increases in putrescine, spermidine and/or spermine, the latter only in the epicarp, were observed in PDJ-treated fruit. MJ induced this behavior only with putrescine in the mesocarp. PpADC transcription was also enhanced soon after the PDJ treatment. Since PDJ-treated fruit were less ripe, their higher PA concentrations in treated fruit are discussed in light of the dual role of these molecules as stress/defense protective compounds and rejuvenating effectors. PMID- 19185953 TI - Topical tar: back to the future. AB - The use of medicinal tar for dermatologic disorders dates back to the ancient times. Although coal tar is utilized more frequently in modern dermatology, wood tars have also been widely employed. Tar is used mainly in the treatment of chronic stable plaque psoriasis, scalp psoriasis, atopic dermatitis, and seborrheic dermatitis, either alone or in combination therapy with other medications, phototherapy, or both. Many modifications have been made to tar preparations to increase their acceptability, as some dislike its odor, messy application, and staining of clothing. One should consider a tried and true treatment with tar that has led to clearing of lesions and prolonged remission times. Occupational studies have demonstrated the carcinogenicity of tar; however, epidemiologic studies do not confirm similar outcomes when used topically. This article will review the pharmacology, formulations, efficacy, and adverse effects of crude coal tar and other tars in the treatment of selected dermatologic conditions. PMID- 19185954 TI - A prospective clinical trial of open-label etanercept for the treatment of hidradenitis suppurativa. AB - BACKGROUND: Medical therapies for hidradenitis suppurativa (HS) are often ineffective. Tumor necrosis factor-alpha inhibitors may be a potential treatment for patients with moderate to severe HS. OBJECTIVES: We sought to evaluate the safety and efficacy of etanercept for patients with severe HS. METHODS: We conducted a phase II clinical trial of etanercept (50 mg/wk subcutaneously) in patients with moderate to severe HS. Efficacy was measured using a Physician Global Assessment and several secondary physician- and patient-reported outcome measures. Responders were classified as those achieving at least a 50% reduction on the Physician Global Assessment score at week 12 compared with baseline. RESULTS: Only 3 of the 15 patients who entered the study were classified as responders (response rate of 20%; 95% confidence interval: 4.3-48.1) based on the intention-to-treat analysis. Dermatology Life Quality Index scores improved slightly from a median of 19 to 15 (P = .02). Comparison of baseline with week-12 Physician Global Assessment scores, and secondary outcome measures of lesion counts and patient pain scores, failed to show statistically significant improvement. Etanercept was generally well tolerated; however, two patients discontinued the study as a result of skin infections at the site of hidradenitis lesions requiring oral antibiotics. LIMITATIONS: Lack of a control group and a small number of participants are limitations. CONCLUSIONS: Our study demonstrated minimal evidence of clinically significant efficacy of etanercept (50 mg/wk subcutaneously) in the treatment of hidradenitis. Future studies using higher doses of etanercept are indicated; however, patients need to be carefully monitored for infection and other adverse events. Randomized, controlled trials will be necessary to demonstrate the risk-to-benefit ratio of tumor necrosis factor-alpha inhibitors in the treatment of hidradenitis. PMID- 19185955 TI - Synthesis, characterisation and in vitro cytotoxicity studies of a series of chiral platinum(II) complexes based on the 2-aminomethylpyrrolidine ligand: X-ray crystal structure of [PtCl2(R-dimepyrr)] (R-dimepyrr=N-dimethyl-2(R) aminomethylpyrrolidine). AB - A series of platinum(II) complexes were synthesised based on the enantiomerically pure amino acid proline. Novel synthetic pathways were developed, adapted from standard peptide chemistry, to produce the 2-aminomethylpyrrolidine (pyrr) ligand and its derivatives with differing arrangements of methyl substituents at the exocyclic amine sites. The crystal structure of [PtCl(2)(R-dimepyrr)] (R dimepyrr=N,N-dimethyl-2(R)-aminomethylpyrrolidine) is reported and the five membered ligand ring has been shown to be in an envelope conformation. Cytotoxicity studies were carried out on the ovarian cancer A2780 tumour cell line and its cisplatin-resistant variant, A2780cisR. Remarkably good activity was seen for several of the drugs when compared to cisplatin despite the addition of substantial steric bulk to the amine groups, and there was a lack of cross resistance with cisplatin seen for some compounds. PMID- 19185956 TI - Physicochemical properties and membrane interactions of anti-apoptotic derivatives 2-(4-fluorophenyl)-3-(pyridin-4-yl)imidazo[1,2-a]pyridine depending on the hydroxyalkylamino side chain length and conformation: an NMR and ESR study. AB - Three imidazo[1,2-a]pyridine derivatives 3a-c have been synthesized from p38 kinase inhibitor structures and evaluated as anti-apoptosis agents. These drugs were designed to interact with nucleic acids and membrane interactions by varying the chain length in position 6, from hydroxyethylamino (3a), to hydroxybutylamino (3b) and hydroxyhexylamino (3c). First experiments showed that 3a and 3b were insoluble in water while 3c could be solubilized in water despite its partition coefficient (logP=3.2). This latter feature was explained by the formation of a fifth intramolecular cycle thus allowing supramolecular structure formation (NMR and MD calculations). The interactions with membranes have been studied using (1)H, (2)H, (31)P Nuclear Magnetic Resonance (NMR), Electron Spin Resonance (ESR) and High Resolution-Magic Angle Spinning (HR-MAS). Despite the insolubility of 3a and 3b in water, these derivatives could be partially solubilized by synthetic phospholipidic model membranes (small unilamellar vesicles, SUV). (1)H NMR paramagnetic broadening experiments performed on the same models showed that 3a was located in the external layer, probably close to the surface while 3b only formed external superficial adducts. Supplementary (31)P, (2)H NMR and ESR experiments on phospholipid dispersions confirmed the location of 3a close to the polar headgroup of the external layer of the membrane, this resulting in a 2K lowering of the transition temperature. Moreover, no significant interaction was detected on the deep part of the layer ((2)H NMR and 16NS ESR experiments). This binding was also found in the presence of cell cultures, as revealed by HR-MAS NMR experiments. Conversely, no significant interaction with membranes was found with 3b or 3c. From both the unexpected solubility of 3c and 3a interactions with membranes, further chemical modifications were finally proposed. PMID- 19185957 TI - A breakfast with alpha-lactalbumin, gelatin, or gelatin + TRP lowers energy intake at lunch compared with a breakfast with casein, soy, whey, or whey-GMP. AB - BACKGROUND & AIMS: Dietary protein plays a role in body weight regulation, partly due to its effects on satiety. The objective was to compare the effects of casein , soy-, whey-, whey without glycomacropeptide (GMP)-, alpha-lactalbumin-, gelatin , or gelatin with tryptophan (TRP)-protein breakfasts at two concentrations on subsequent satiety and energy intake (EI). METHODS: Twenty-four healthy subjects (mean+/-SEM BMI: 24.8+/-0.5 kg/m(2); age: 25+/-2 years) received a breakfast; a custard with casein, soy, whey, whey-GMP, alpha-lactalbumin, gelatin, or gelatin+TRP as protein source with either 10/55/35 (normal) or 25/55/20 (high) En% protein/carbohydrate/fat in a randomized, single-blind design. At the precedingly determined time point for lunch, 180 min, subjects were offered an ad lib lunch. Appetite profile (Visual Analogue Scales, VAS) and EI were determined. RESULTS: Both at the level of 10 and 25 En% from protein, EI at lunch was approximately 20% lower after an alpha-lactalbumin or gelatin (+TRP) breakfast (2.5+/-0.2 MJ) compared with after a casein, soy, or whey-GMP breakfast (3.2+/ 0.3 MJ, p<0.05). Appetite ratings at 180 min differed 15-25 mm (approximately 40%, p<0.05) between types of protein. Differences in EI were a function of differences in appetite ratings (R(2)=0.4, p<0.001). CONCLUSIONS: Different proteins (alpha-lactalbumin, gelatin, gelatin+TRP) that are approximately 40% more satiating than other proteins (casein, soy, whey, whey-GMP) induce a related approximately 20% reduction of subsequent energy intake. PMID- 19185959 TI - Internal loads in the human tibia during gait. AB - BACKGROUND: Internal loads in long bones are of special interest, for testing and optimization of current implant designs for fracture treatment. Inverse dynamic musculoskeletal models were successfully used to determine the muscle forces in the lower extremities, however the internal loads expressed as forces and moments along the human tibia during gait have not been reported. METHODS: A musculoskeletal model of the lower extremities, developed and provided under public domain from the AnyBody research group (www.anybody.aau.dk), was modified to determine the three-dimensional internal loads along the tibial axis during gait. To corroborate this numerical model, the calculated resultant hip contact force as well as the axial force on the tibial plateau were compared to in vivo data from the literature. FINDINGS: The highest internal loads were the axial force with up to 4.7 bodyweight and the bending moment in the sagittal plane with up to 71.6 bodyweight times millimetre in the late stance phase. The extreme values of the internal loads along the tibial axis varied up to 1.5 bodyweight and 85.7 bodyweight times millimetre. In the distal part of the tibia, the axial force dominated the three-dimensional internal load case, whereas the internal moments became more significant with increasing distance from the ankle joint. INTERPRETATION: This study provides for the first time the three-dimensional internal loads, expressed as forces and moments along the human tibia during normal gait. The results of this study could be used to improve the mechanical behaviour of current implant designs for the treatment of tibial fractures to avoid implant failures under in vivo loading conditions. PMID- 19185958 TI - Management of the third stage of labour--focus group discussions with Swedish midwives. AB - OBJECTIVE: to explore Swedish midwives' experiences of management of third stage of labour. DESIGN: six focus group discussions were performed and the analysis was based on content analysis. SETTING: the midwives worked at six hospitals: three university hospitals and three provincial hospitals located from the south west to the north of Sweden. PARTICIPANTS: 32 midwives with extensive experience of assisting women in childbirth. FINDINGS: the analysis generated three categories: 'bring the process under control', 'protect normality and women's birthing experiences' and 'maintain midwives' autonomy'. This study demonstrates that management of the third stage of labour varies greatly. Not all midwives were convinced that administration of prophylactic oxytocin in the third stage of labour was always the best alternative for all women who had a normal birth. KEY CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: the midwives exhibited self-confidence in evaluating the physiological process, and endeavoured to leave the physiological process undisturbed if no other risks were apparent. Their decisions concerning third stage management were based on a combination of previous experience, hospital guidelines, risk assessment and sensitivity to each woman's needs. This study demonstrates that management of the third stage of labour varies greatly. The findings show the importance of reaching a balance between treating birth as a normal process and as a biomedical event. PMID- 19185960 TI - Biomechanics of latissimus dorsi transfer for irreparable posterosuperior rotator cuff tears. AB - BACKGROUND: Latissimus dorsi transfer is the treatment most frequently used for restoring function in shoulders with irreparable posterosuperior rotator cuff tears. Yet, functional outcomes of the transfers are unpredictable and vary among patients. METHODS: A three-dimensional upper-extremity computational model was used to simulate and analyze the biomechanical consequences of transferring the latissimus dorsi to four attachment sites: the infraspinatus, supraspinatus, subscapularis and teres minor insertions. Functions of a normal shoulder were simulated, as well as those and of a shoulder with a posterosuperior rotator cuff tear before and after muscle transfers were simulated. Parameters such as active and passive moment-generating capacity, and the moment arm and fiber excursion ratio of the transferred muscle were analyzed. FINDINGS: All muscle transfers resulted in a large increase in shoulder external rotation strength. The latissimus dorsi was an external rotator after the transfer, but the fiber excursion ratio decreased accordingly. When the latissimus dorsi was transferred to the infraspinatus, supraspinatus or subscapularis insertion, it changed from extensor to flexor at the beginning of flexion. The flexion moment arm of the latissimus dorsi after the transfers was generally decreased. Shoulder abduction strength did not improve. Decrease in fiber excursion ratio during abduction and flexion was observed after the transfer. Side effects of the muscle transfers, such as the reduction of active adduction, extension and internal rotation of the shoulder, were explored. INTERPRETATION: A transfer to teres minor insertion was not recommended. Infraspinatus insertion was found to be a preferred attachment site in latissimus dorsi transfer, provided that the patient had a strong deltoid. PMID- 19185961 TI - Passive mechanical properties of gastrocnemius in people with multiple sclerosis. AB - BACKGROUND: There is evidence to suggest that contractile properties of muscles in people with multiple sclerosis change as a consequence of demyelination in central nervous system. However, passive properties of muscles in people with multiple sclerosis have not been previously investigated. The purpose of this study was to characterise passive mechanical properties of gastrocnemius in people with multiple sclerosis and to compare these properties with those of gastrocnemius in neurologically normal people. METHODS: Ten people with multiple sclerosis having signs and symptoms of weakness in the legs (Disease step 1-3) and 10 age- and sex-matched healthy people participated in the study. Ultrasound images of muscle fascicles of medial gastrocnemius as well as passive ankle torque and ankle angle data were obtained simultaneously as the ankle was rotated through its full range with the knee in a range of positions. Analysis of ultrasound images and passive ankle torque-angle relations allowed us to derive the slack lengths and maximal strains of whole muscle-tendon units, muscle fascicles and tendons. Paired-samples t-tests were used to compare these variables in the two groups. RESULT: There was no difference between subjects with multiple sclerosis and healthy controls in the mean slack lengths and mean maximal strains of the whole muscle-tendon units or of their fascicles or tendons. INTERPRETATIONS: These data suggests that typically, in people with multiple sclerosis who have impaired lower limbs but are still ambulatory, the passive mechanical properties of the gastrocnemius muscles are normal. PMID- 19185962 TI - Estimating the spatial scale of herbicide and soil interactions by nested sampling, hierarchical analysis of variance and residual maximum likelihood. AB - An unbalanced nested sampling design was used to investigate the spatial scale of soil and herbicide interactions at the field scale. A hierarchical analysis of variance based on residual maximum likelihood (REML) was used to analyse the data and provide a first estimate of the variogram. Soil samples were taken at 108 locations at a range of separating distances in a 9 ha field to explore small and medium scale spatial variation. Soil organic matter content, pH, particle size distribution, microbial biomass and the degradation and sorption of the herbicide, isoproturon, were determined for each soil sample. A large proportion of the spatial variation in isoproturon degradation and sorption occurred at sampling intervals less than 60 m, however, the sampling design did not resolve the variation present at scales greater than this. A sampling interval of 20-25 m should ensure that the main spatial structures are identified for isoproturon degradation rate and sorption without too great a loss of information in this field. PMID- 19185963 TI - Bacterial toxicity comparison between nano- and micro-scaled oxide particles. AB - Toxicity of nano-scaled aluminum, silicon, titanium and zinc oxides to bacteria (Bacillus subtilis, Escherichia coli and Pseudomonas fluorescens) was examined and compared to that of their respective bulk (micro-scaled) counterparts. All nanoparticles but titanium oxide showed higher toxicity (at 20 mg/L) than their bulk counterparts. Toxicity of released metal ions was differentiated from that of the oxide particles. ZnO was the most toxic among the three nanoparticles, causing 100% mortality to the three tested bacteria. Al(2)O(3) nanoparticles had a mortality rate of 57% to B. subtilis, 36% to E. coli, and 70% to P. fluorescens. SiO(2) nanoparticles killed 40% of B. subtilis, 58% of E. coli, and 70% of P. fluorescens. TEM images showed attachment of nanoparticles to the bacteria, suggesting that the toxicity was affected by bacterial attachment. Bacterial responses to nanoparticles were different from their bulk counterparts; hence nanoparticle toxicity mechanisms need to be studied thoroughly. PMID- 19185964 TI - "Race" and "ethnicity" in biomedical research: how do scientists construct and explain differences in health? AB - Social and biomedical scientists, journal editors, and public health officials continue to debate the merits of the use of race and ethnicity in health-related research. As biomedical research focuses on issues of racial or ethnic health disparities, it remains unclear how biomedical scientists investigate race or ethnicity and health. This paper examines how biomedical researchers construct and analyze race or ethnicity in their studies and what conclusions they make about difference and health. Using content analysis of 204 biomedical research journal publications, which were supported by grants won from the National Cancer Institute of the National Institutes of Health in the USA, I demonstrate that although authors tended to see race or ethnicity as important and significant in their research, they rarely defined or operationalized the concepts adequately. Moreover, when presenting findings of racial or ethnic difference, authors generally did not provide explanations of the difference. I argue that this under theorized and unspecified use of race or ethnicity and the biological conclusions drawn about health and difference have the potential to reify "race" and to limit our thinking about what these biomedical differences suggest about health disparities and inequalities in general. PMID- 19185966 TI - An SEM evaluation of conditioned and bonded enamel following carbamide peroxide bleaching and casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) treatment. AB - OBJECTIVES: The study aimed to evaluate, (a) the surface morphology of acid etched/conditioned enamel following carbamide peroxide bleaching with/without casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) (Tooth Mousse (MI Paste); GC Corp., Tokyo, Japan) treatment; and b) the nature of the bonded resin enamel interfaces formed with a self-etching primer adhesive. METHODS: Twenty four human adult molars were each sectioned into four, the specimens divided and treated according to four experimental groups: 1, no treatment; 2, 16% carbamide peroxide bleaching; 3, CPP-ACP paste; 4, bleaching and CPP-ACP paste. A self etching primer adhesive (Clearfil SE Bond, CSE) was used. The specimens were further divided into four subgroups for etching/conditioning: A, CSE Primer only; B, 30-40% phosphoric acid and CSE primer; C, 15% EDTA and CSE primer; D, 20% polyacrylic acid and CSE primer. Bonded specimens were also prepared. The morphology of the etched/conditioned enamel surfaces and polished resin-enamel interfaces of the bonded specimens were observed by field-emission scanning electron microscopy (FE-SEM). RESULTS: Treatment with CPP-ACP did not inhibit phosphoric acid etching of enamel. Poorly defined enamel etch patterns were observed with the other conditioners in all the groups. The morphology of the bonded resin-enamel interfaces observed on FE-SEM for each acid etching/conditioning subgroup was similar in all the experimental groups, except after bleaching. Resin infiltration into enamel was observed with prior phosphoric acid etching and polyacrylic acid conditioning and was unaffected by experimental group treatments. CONCLUSIONS: The use of a CPP-ACP paste with or without prior bleaching did not inhibit enamel etching. Enamel etching/conditioning may help improve bonding efficiency of the self-etching primer adhesive after CPP-ACP treatment. PMID- 19185965 TI - Violence from parents in childhood and obesity in adulthood: using food in response to stress as a mediator of risk. AB - Guided by a life course perspective and concepts from models of stress and coping, this study tested the extent to which self-reported profiles of physical and psychological violence in childhood from parents were associated with greater odds of obesity in adulthood. This study also examined the extent to which adults' greater use of food in response to stress served as a mediator of potential associations of risk. Multivariate regression models were estimated using data from 1650 respondents in the 1995-2005 National Survey of Midlife in the U.S. (MIDUS). Results indicated that respondents who reported having experienced both psychological and physical violence from parents-with at least one type of violence having reportedly occurred frequently-were more likely to be classified as obese in contrast to respondents who reported never having experienced either type of violence from parents. Evidence from a sequence of models that tested mediation effects indicated that greater use of food in response to stress among respondents with problematic histories of violence explained, in part, their higher risk of adult obesity. Findings contribute to the growing body of evidence regarding psychosocial predictors of obesity, as well as the physical health consequences of childhood family violence. Results further suggest the importance of addressing particular aspects of life course social relationships-such as violence in childhood from parents-and their implications for psycho-behavioral uses of food within efforts to reduce rates of adult obesity. PMID- 19185967 TI - The approach to delayed resuscitation in paediatric cardiac arrest: A survey of paediatric intensivists in Canada. AB - AIM: To determine how long a period of having had no cardiopulmonary resuscitation (CPR) (delay time) is considered to result in subsequent futile efforts at resuscitation. METHODS: In 2007 a survey was mailed to all 77 paediatric intensivists in Canada. Three scenarios of witnessed cardiac arrest were presented: out-of-hospital, in-hospital, and in-hospital with extracorporeal CPR (E-CPR). Each scenario asked what delay time would make attempts at resuscitation futile for survival to hospital discharge, and for survival to hospital discharge in a better than vegetative state. Comparisons of median [inter-quartile range] used Wilcoxon-signed-rank or Friedman tests with Bonferroni corrections. RESULTS: The response rate was 49/77 (64%). The delay time was significantly different between rhythms within all scenarios (p<.001); and was significantly shorter for survival than for better than vegetative survival (p<.006) except when E-CPR was to be used. The delay time was not significantly different between the in-hospital and out-of-hospital scenario with the same rhythms (p>.01). The delay time was significantly shorter in scenarios with asystole versus pulseless electrical activity with (p=.010) or without (p<.001) an arterial line with absent pulsation. In out-of-hospital arrest, the delay time for survival varied from 15 [10-20]min for asystole to 20 [15-20]min for pulseless electrical activity. In in-hospital scenarios, the delay time for survival varied from 10 [10-20]min for asystole, to 15 [10-20]min for most other rhythms. CONCLUSION: A delay time of 15 [10-20] (range 5-30)min was considered futile for survival. This has implications for pronouncing death in donation after cardiac death. PMID- 19185968 TI - Patient suitability for magnetic resonance guided focused ultrasound surgery of uterine fibroids. AB - OBJECTIVE: To assess selection criteria used to determine eligibility for magnetic resonance-guided focused ultrasound surgery (MRgFUS) for the treatment of symptomatic uterine fibroids; to assess the percentage of patients suitable for MRgFUS. STUDY DESIGN: A retrospective analysis of 144 patients seeking minimally invasive treatment options for symptomatic uterine fibroids at a single treatment center. Clinical eligibility for MRgFUS was assessed at a gynecology clinic by a Gynecology research fellow trained in the procedure and suitability was assessed by magnetic resonance imaging. Several techniques were used to mitigate against factors that are contra-indications for MRgFUS. RESULTS: 100% of patients interested in MRgFUS were deemed clinically eligible for the procedure and 74% were deemed technically suitable to proceed with treatment. CONCLUSIONS: Mitigation techniques allow for less restrictive MRgFUS selection criteria for treatment for symptomatic uterine fibroids. These less restrictive criteria are expected to expand the pool of patients for whom MRgFUS is a viable treatment option for uterine fibroid symptoms. PMID- 19185969 TI - The effect of extracellular adenosine triphosphate on the spontaneous contractility of human myometrial strips. AB - OBJECTIVES: Extracellular ATP is involved in cell-cell signalling in a variety of tissues but its effects in tissue level signalling in the myometrium have been poorly studied to date. We hypothesised that extracellular ATP was involved in the control of myometrial contractile frequency and/or force. STUDY DESIGN: In vitro study of the effect of altering the concentration of extracellular ATP on the spontaneous contractility of human myometrial strips obtained from term elective caesarean sections. RESULTS: Decreasing extracellular ATP levels by the ectoATP-ase agent apyrase VI (1-50 units/mL) produced a dose dependent decrease in contractile frequency (decrease of 46.3% compared with the baseline frequency at 20 units/mL, p=0.001, n=6). Contractility was unchanged by apyrase VII (20 units/mL), an agent with relatively greater ADPase activity, indicating an effect via ATP and not ADP. Contractile frequency increased after addition of ATP 10-100 microM (an increase to 145.8% of baseline frequency at 100 microM: 126.1-165.5%, p=0.005, n=7) or the ATPase inhibitor ARL at 100 microM (an increase to 136.3% of the baseline frequency: 107.1-165.5, p=0.03, n=7). Contractile force remained unchanged by these agents. CONCLUSIONS: Extracellular ATP shows a dose-response relationship to contractile frequency but does not affect contractile force. Consequently it may be involved in the pacemaking mechanism for the generation of uterine contractions. PMID- 19185970 TI - Internalization of agri-environmental policies and the role of institutions. AB - Recent investigations have indicated that environmental and conservation policies frequently fail to reach anticipated aims, which raises concern over the cost effectiveness of governmental policy-related expenditure. The limited effectiveness of policies is often attributed to methodological aspects of policy implementation. However, an alternative reason can be the limited internalization of policies, so that these are only implemented to a minimum level to attain benefits or avoid penalties. It is postulated that increased internalization of policies can considerably improve their effectiveness and that suitable institutional arrangements exist to support such increases in internalization. In this paper, we review the available literature on internalization and its institutional aspects, and propose a framework, based on self-determination theory, for evaluation of potential internalization by farm managers that are expected to implement policies. This method was applied to a small case study for dairy farmers in the nationally important landscape of the Northern Friesian Woodlands, The Netherlands. The results showed that organic farmers were internally motivated for nature conservation and had strong institutional links. They were more likely to internalize the goals of environmental policy schemes than conventional farmers who focused predominantly on financial rewards. We suggest that policy developers promote internalization of policies by tuning policy instruments to the specificities of farmers' motivations to preserve farmers' nature and to focus on institutional support to help internalization. PMID- 19185971 TI - A bioeconomic model of afforestation in Southern Ontario: integration of fiber, carbon and municipal biosolids values. AB - This study explores the economic attractiveness of afforestation as a strategy for the joint production of fibre, carbon storage and biosolids (treated municipal sewage sludge) disposal for municipalities in Southern and Central Ontario, Canada. We use a spatial, stochastic model, the Canadian Forest Service Afforestation Feasibility Model (CFS-AFM), to simulate a range of spatial biosolids application scenarios in hybrid poplar afforestation projects. Results suggest that such joint afforestation strategies could be financially attractive. Significant cost savings can be expected through decreases in transportation distances and avoided waste disposal fees. Sensitivity analysis is used to examine the effects of variations in critical model parameters on net present values. Our findings indicate that waste disposal savings from application of biosolids on hybrid poplar plantations combined with incentives for landowners to sequester carbon can easily compete with agricultural land rental values in some regions of Ontario. Social acceptance of this kind of activity, however, may be an impediment to adoption. PMID- 19185972 TI - 3D freehand ultrasound for in vivo determination of human skeletal muscle volume. AB - Skeletal muscle volume is an important indicator of muscle function. Three dimensional (3D) freehand ultrasound provides a noninvasive method for determining muscle volume and is acquired using a standard clinical ultrasound machine and an external tracking system to monitor transducer position. Eleven healthy volunteers were scanned with a 3D freehand system that uses an optical tracking device. Interest was concentrated on one of the muscles of the quadriceps group, rectus femoris and volume measurements performed on 30 mm cross sections were compared with measurements derived from magnetic resonance imaging. Measured muscle volumes ranged from 5 cm(3) to 28 cm(3). The mean difference between measurements from 3D freehand ultrasound and magnetic resonance was 0.53 cm(3) with 95% limits of agreement of +/-2.14 cm(3). Muscle volume measurements obtained using 3D ultrasound were within +/-16% of the corresponding value from magnetic resonance imaging. We have shown for the first time that 3D freehand ultrasound can be used to determine human skeletal muscle volume accurately in vivo. PMID- 19185973 TI - Effect of transosseous application of low-intensity ultrasound at the tendon graft-bone interface healing: gene expression and histological analysis in rabbits. AB - The present study investigates the effect of transosseous low-intensity pulsed ultrasound (LiUS) on the healing at tendon graft-bone interface, in molecular and histological level. The anterior cruciate ligament (ACL) in both knees of 52 New Zealand White rabbits was excised and replaced with the long digital extensor. A custom-made ultrasound transducer was implanted onto the medial tibial condyle, adjacent to the surface of the bone tunnel at both knees of the rabbits. The LiUS treated right knees received 200-mus bursts of 1 MHz sine waves at a pulse repetition rate of 1 kHz and with 30 mW/cm(2) spatial-average temporal-average intensity for 20 min daily (study group), while the left knee received no LiUS (control group). Thirty-six rabbits were used to perform semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis from both study and control groups for transforming growth factor-beta1 (TGF-beta1), biglycan and collagen I. RT-PCR products showed statistically significant upregulation of biglycan and collagen I gene expression in the study group, while TGF-beta1 gene expression exhibited a bimodal profile. Histological examination performed in 16 rabbits from both groups supported the findings of the molecular analysis, indicating a faster healing rate and a more efficient ligamentization process after ultrasound treatment. These findings suggest that transosseous application of LiUS enhances the healing rate of the tendon graft-bone interface, possibly by affecting the expression levels of genes significant for the tendon to bone healing process. PMID- 19185974 TI - Ultrasound Doppler monitoring of soft tissues in vitro and tissue phantoms heating and thermal destruction induced by acoustic remote palpation. AB - The rise of shear strain value under temperature increase in biological tissue samples in vitro and tissue phantoms was studied and the range of shear modulus and viscosity calculated. It has been shown that the acoustic radiation force based methods with the usage of ultrasound Doppler probing provides the potential ability of noninvasive real-time monitoring of tissues' ultrasound thermal destruction process. At that, the thermal destruction is possible under action of wave beam that creates the radiation force and local tissue displacements so that tissue ablation and acoustic remote palpation could be realized by means of the same ultrasound transducer. The experiments were performed using gelatin-based tissue-mimicking phantoms and freshly excised samples of bovine muscle tissue. It was determined also that fluctuating pattern of detected displacement amplitude variation is the indicator of the phase transitions beginning in the heated field of soft tissue or tissue phantom. PMID- 19185975 TI - Four-year outcome of a prospective randomised trial comparing bipolar plasmakinetic and monopolar transurethral resection of the prostate. AB - BACKGROUND: No data have been published on the midterm efficacy of bipolar transurethral resection of the prostate (TURP). OBJECTIVE: To evaluate 4-yr results from a prospective randomised trial comparing bipolar TURP with standard monopolar TURP. DESIGN, SETTING, AND PARTICIPANTS: Seventy patients with symptomatic benign prostatic hyperplasia were enrolled in this prospective randomised controlled trial in a tertiary-care institution. Inclusion criteria were age > 50 yr, good performance status, urinary retention, International Prostate Symptom Score (IPSS) > or = 18, and maximal flow rate (Q(max)) < or = 15 ml/s. Exclusion criteria were prostate volume < 30 cm(3), documented or suspected prostate cancer, neurogenic bladder, bladder stone or diverticula, urethral stricture, and maximal bladder capacity > 500 ml. INTERVENTION: Patients underwent standard or bipolar plasmakinetic TURP performed by the same surgeon using the same surgical technique. MEASUREMENTS: Treatment efficacy was evaluated at 1, 2, 3, and 4 yr by comparing urinary flow rates, IPSS, and estimated postvoid residual (PVR) urine volume. Midterm complications were also recorded. RESULTS AND LIMITATIONS: The number of dropouts was not statistically significantly different in the two groups (p=0.2). The significant improvements in both groups were maintained at 4 yr for the IPSS, quality of life score, Q(max), and PVR versus baseline values. The main outcome variables at 4 yr for bipolar and monopolar TURP were mean IPSS 6.9 and 6.4 (p=0.58); mean Q(max) 19.8 ml/s and 21.2 ml/s (p=0.44), and mean PVR volume 42 ml and 45 ml (p=0.3). Overall, 2 of 32 (6.2%) and 3 of 31 (9.6%) patients required reoperation because of late complications (p=0.15). The major study limitation was the small sample size. CONCLUSIONS: This study represents the secondary, midterm analysis of a previously published trial. Our 4-yr data confirm our initial positive findings for the efficacy and safety of bipolar plasmakinetic TURP. Larger well-designed studies are needed to corroborate these findings. PMID- 19185976 TI - Effects of phosphodiesterase inhibitors on the inflammatory response of endothelial cells stimulated by myeloperoxidase-modified low-density lipoprotein or tumor necrosis factor alpha. AB - BACKGROUND: Sildenafil, vardenafil, and tadalafil are phosphodiesterase type 5 inhibitors (PDE5-Is) usually used in the treatment of erectile dysfunction (ED). Previously, we have shown the presence of myeloperoxidase-modified low-density lipoprotein (Mox-LDL) in the penises of patients with ED, and we have shown the impact of Mox-LDL on cyclic monophosphate (cGMP) level. In vitro, Mox-LDL triggered the inflammatory response by increasing the release of both interleukin 8 (IL-8) and tumor necrosis factor alpha (TNF-alpha) by endothelial cells (ECs) and monocytes respectively. OBJECTIVE: To determine whether or not the three therapeutically PDE5-Is protect against the proinflammatory effects of Mox-LDL or TNF-alpha on ECs. DESIGN, SETTING, AND PARTICIPANTS: ECs (EA.hy926) were incubated in the presence of either TNF-alpha (100 pg/ml) or Mox-LDL (200 microg/ml) with each of the three PDE5-Is (1 microM, 5 microM, and 10 microM) respectively. IL-8 production was measured in the supernatant after 48 h of incubation. MEASUREMENTS: All experiments were repeated at least three times. Statistical analysis was performed with an ANOVA. RESULTS AND LIMITATIONS: Two way ANOVA analysis showed that TNF-alpha alone (p<0.001) or Mox-LDL alone (p<0.001) increased IL-8 production. Sildenafil, vardenafil, or tadalafil alone did not generate an increase of IL-8 production. Tadalafil in combination with Mox-LDL and TNF-alpha showed a decrease of IL-8 (p<0.05) compared with sildenafil and vardenafil. CONCLUSIONS: Among the three available PDE5-Is, tadalafil showed an additional potentially anti-inflammatory effect on relaxation. Those data could be considered for the chronic use of PDE5-Is, but extrapolations of experimental evidence to the clinical setting should be made cautiously. PMID- 19185977 TI - Retropubic, laparoscopic, and robot-assisted radical prostatectomy: a systematic review and cumulative analysis of comparative studies. AB - CONTEXT: Despite the wide diffusion of laparoscopic radical prostatectomy (LRP) and robot-assisted laparoscopic radical prostatectomy (RALP), only few studies comparing the results of these techniques with the retropubic radical prostatectomy (RRP) are currently available. OBJECTIVE: To evaluate the perioperative, functional, and oncologic results in the comparative studies evaluating RRP, LRP, and RALP. EVIDENCE ACQUISITION: A systematic review of the literature was performed in January 2008, searching Medline, Embase, and Web of Science databases. A "free-text" protocol using the term radical prostatectomy was applied. Some 4000 records were retrieved from the Medline database; 2265 records were retrieved from the Embase database;, and 4219 records were retrieved from the Web of Science database. Three of the authors reviewed the records to identify comparative studies. A cumulative analysis was conducted using Review Manager software v.4.2 (Cochrane Collaboration, Oxford, UK). EVIDENCE SYNTHESIS: Thirty-seven comparative studies were identified in the literature search, including a single, randomised, controlled trial. With regard to the perioperative outcome, LRP and RALP were more time consuming than RRP, especially in the initial steps of the learning curve, but blood loss, transfusion rates, catheterisation time, hospitalisation duration, and complication rates all favoured LRP. With regard to the functional results, LRP and RRP showed similar continence and potency rates. Similarly, no significant differences were identified between LRP and RALP, while a single, nonrandomised, prospective study suggested advantages in terms of both continence and potency recovery after RALP, compared with RRP. With regard to the oncologic outcome, LRP and RALP were associated with positive surgical margin rates similar to those of RRP. CONCLUSIONS: The quality of the available comparative studies was not excellent. LRP and RALP are followed by significantly lower blood loss and transfusion rates, but the available data were not sufficient to prove the superiority of any surgical approach in terms of functional and oncologic outcomes. Further high quality, prospective, multicentre, comparative studies are needed. PMID- 19185978 TI - Editorial comment on: Four-year outcome of a prospective randomised trial comparing bipolar plasmakinetic and monopolar transurethral resection of the prostate. PMID- 19185979 TI - Editorial comment on: Effects of phosphodiesterase inhibitors on the inflammatory response of endothelial cells stimulated by myeloperoxidase-modified low-density lipoprotein or tumor necrosis factor alpha. PMID- 19185980 TI - Editorial comment on: Four-year outcome of a prospective randomised trial comparing bipolar plasmakinetic and monopolar transurethral resection of the prostate. PMID- 19185981 TI - Functional magnetic resonance imaging in prostate cancer. AB - CONTEXT: Magnetic resonance imaging (MRI) combined with magnetic resonance spectroscopy imaging (MRSI), dynamic contrast-enhanced MRI, and diffusion weighted MRI emerged as promising tests in the diagnosis of prostate cancer, and they show encouraging results. OBJECTIVE: This review emphasizes different functional MRI techniques in the diagnosis of prostate cancer and includes information about their clinical value and usefulness. EVIDENCE ACQUISITION: The authors searched the Medline, Embase, and Cochrane Library databases. There were no language restrictions. The last search was performed in October 2008. EVIDENCE SYNTHESIS: The combination of conventional MRI with functional MRI techniques is more reliable for differentiating benign and malignant prostate tissues than any other diagnostic procedure. At present, no guideline is available that outlines which technique is best in a specific clinical situation. It also remains uncertain whether improved spatial resolution and signal-to-noise ratio of 3-T MRI will improve diagnostic performance. CONCLUSIONS: A limited number of small studies suggest that functional MRI may improve the diagnosis and staging of prostate cancer. This finding needs further confirmation in larger studies, and cost-effectiveness needs to be established. PMID- 19185982 TI - Optic neuritis and palatal dysarthria as presenting features of post-infectious GQ1b antibody syndrome. AB - A 31-year-old man had optic neuritis 2 weeks after a diarrheal illness, followed by several deficits including palatal dysarthria, diplopia, ataxia, sensory dysfunction, and mild dysautonomia. Brain MRI and CSF were normal. Nerve conduction studies were initially normal and subsequently showed mild reduction in sensory amplitudes. Anti-GQ1b IgG titer was positive. Deficits resolved after treatment with IVIg. This clinical constellation represents an overlap between Miller Fisher syndrome (MFS) and the pharyngeal-cervical-brachial (PCB) variant of Guillain-Barre syndrome (GBS), along with the infrequently reported central feature of optic neuritis. Campylobacter jejuni enteritis may have triggered the syndrome by molecular mimicry. GQ1b antibodies are associated with MFS, GBS, Bickerstaff brainstem encephalitis and PCB; they form an overlapping spectrum of features, hence the anti-GQ1b syndrome. PMID- 19185983 TI - A modern approach to CSF analysis: pathophysiology, clinical application, proof of concept and laboratory reporting. AB - The CNS immune response often leads to characteristic interrelated biochemical changes in cerebrospinal fluid. Multiple analytes, i.e. cell count, cell differential, evaluation of barrier function and intrathecal IgG, IgA and IgM synthesis should be included in basic diagnostic workup. We describe the scientific background, laboratory investigations and characteristic patterns found with basic CSF analysis, based on the recommendations of the German cerebrospinal fluid society. The concept is substantiated by retrospectively analyzing data of 4026 paired CSF/serum samples. 53% of our samples presented with at least one or several combined abnormal findings. An intrathecal IgG, IgA or IgM immunoglobulin response (37%, n=1481) and a blood-CSF barrier dysfunction (37%; n=1473) were most frequent; followed by an elevated leukocyte cell count (25%; n=992). The immunoglobulin response showed an intrathecal production of IgG in 49% (n=731/1481), which was only detectable in isoelectric focusing in 27% (n=200/731). Intrathecal IgM (n=389) and IgA (n=361) synthesis presented with nearly equal frequency of 25% in samples with intrathecal immunoglobulin response. The immunoglobulin pattern showed a solitary one class reaction of IgG, IgA or IgM in 67%, a combined two class reaction of IgG/IgA, IgG/IgM or IgA/IgM synthesis in 16% and a combined three-class reaction of IgG, IgA and IgM in 17%. This approach generates valuable but numerous complex and interrelated biochemical data. We therefore developed a knowledge-based system combined with visual oriented laboratory output to transfer the information more effectively. This often uncovers typical patterns specific for distinct neurological diseases, is well accepted by our medical community documented by a 37% increase in external ordering. PMID- 19185984 TI - Nerve root prolapse into a spinal arachnoid cyst--an unusual cause of radiculopathy. AB - Arachnoid cysts are rare lesions of the spine and can present with myelopathy, radiculopathy, local pain or a combination of these symptoms. Nerve root prolapse into an arachnoid cyst causing radiculopathy has not been reported before. We report a nerve root prolapse into a spinal arachnoid cyst presenting clinically as radiculopathy. An 18-year-old female patient presented with mid-back pain, right anterior thigh pain and hip flexor weakness. Magnetic resonance imaging (MRI) and computerized tomography (CT) myelography revealed an arachnoid cyst at T12-L1 level on the right side. At surgery, a nerve root was seen prolapsing into an extradural arachnoid cyst. The nerve root was replaced back into dural sac and the dural defect closed. At 20 months of follow-up, the patient continues to be asymptomatic with no evidence of recurrence on imaging. Replacing the prolapsed nerve root into the dural sac with meticulous closure of the dural defect could lead to good clinical outcome. We propose a modification to the popular classification of these lesions to better rationalize their surgical management. Classification of extradural arachnoid spinal cysts (Nabors's type 1) should be based on the presence or absence of dural communication. Sacral meningoceles (Nabors' type 1B) should be excluded from the classification as they have free communication with the thecal sac and are not true spinal cysts. PMID- 19185985 TI - Relationship of homocysteine with other risk factors and outcome of ischemic stroke. AB - OBJECTIVE: Hyperhomocysteinemia (HH) is an emerging risk factor for ischemic stroke but its role in outcome is controversial. We compare the risk factors, nature of stroke and outcome of patients with and without hyperhomocysteinemia. PATIENTS AND METHODS: CT proven ischemic stroke patients were included. The conventional risk factors such as diabetes, hypertension, hyperlipidemia, obesity, smoking, and family history of stroke were recorded. Dietary history was noted. Fasting serum homocysteine (Hcy), B12 and folic acid were estimated after 1 month of stroke. Severity of stroke was assessed by Canadian Neurological Scale (CNS) and outcome at 3 months by Barthel Index (BI) score into good (BI > or = 12) and poor (BI < 12). Serum Hcy, B12 and folic acid were also estimated in 200 normal healthy volunteers. RESULTS: There were 198 patients with ischemic stroke whose median age was 56 years and 36 were females. In the study group, 41.4% patients were vegetarian, 55.1% hypertensive, 24.7% diabetic, 30.8% smoker, 61.1% sedentary and 28.8% obese. 23.2% had past history of stroke and 21.7% had stroke in their first degree relative. Serum cholesterol was elevated in 11.7% and LDL in 10.8% patients. Serum Hcy was elevated in 60.6% and serum B12 low in 25.7% and folic acid in 42.1%. Hcy levels correlated with serum B12 and LDL. Patients with hyperhomocysteinemia had significantly better outcome at 3 months. Hcy levels in stroke patients did not significantly differ from controls. CONCLUSION: Hyperhomocysteinemia is found in 60.6% stroke patients, which is related to low serum B12 level. Patients with hyperhomocysteinemia had a better 3-month outcome. PMID- 19185986 TI - Histone deacetylase inhibitor induced modulation of anti-estrogen therapy. AB - Histone deacetylase (HDAC) inhibitors are a novel class of anti-tumor agents with a potential role in the treatment of breast cancer. In ER-positive cells, treatment with selective and non-selective HDAC inhibitors has been associated with a transcriptional down-regulation (and possibly protein modification via the HSP90 chaperone function) of ER and its response genes. In ER-negative cell lines, HDAC inhibitors have been shown to re-establish ER expression. In addition, HDAC inhibitors have been reported to modulate the progesterone receptor. Despite the opposing effects in ER-positive and ER-negative breast cancer cells, the addition of an HDAC inhibitor potentiated and restored the efficacy of anti-estrogen therapy in preclinical models. This has led to the initiation of several clinical trials combining HDAC inhibitors with anti estrogen therapy. In this review, we will summarize the relationship between estrogen signaling and HDACs, examine how HDAC inhibitors impact this relationship and synergize with anti-estrogens to inhibit tumor growth, and discuss the clinical possibilities and potential of this new approach. PMID- 19185988 TI - Side population in oral squamous cell carcinoma possesses tumor stem cell phenotypes. AB - To characterize the biological features of side population (SP) in oral squamous cell carcinoma (OCC), SP and non-SP were sorted and compared. The SP cells were more clonogenic and in nude mice, only 10,000 SP cells were needed for tumor development compared to 1,000,000 non-SP cells. The SP cells expressed higher levels of ABCG2, ABCB1, CD44, Oct-4, Bmi-1, NSPc1 and CK19. The SP cells generated SP and non-SP populations, whereas the non-SP cells generated only non SP. These findings provide the first evidence that SP in OCC possesses tumor stem cell phenotypes and may play an important role in OCC tumorigenesis. PMID- 19185989 TI - Thermal radiation of di-tert-butyl peroxide pool fires-Experimental investigation and CFD simulation. AB - Instantaneous and time averaged flame temperatures T , surface emissive power SEP and time averaged irradiances E of di-tert-butyl peroxide (DTBP) pool fires with d=1.12 and 3.4m are investigated experimentally and by CFD simulation. Predicted centerline temperature profiles for d=1.12m are in good agreement with the experimental emission temperature profiles for x/d>0.9. For d=3.4m the CFD predicted maximum centerline temperature at x/d=1.4 is 1440 K whereas the emission temperature experimentally determined from thermograms at x/d approximately 1.3 is 1560 K. The predicted surface emissive power for d=1.12m is 115 kW/m(2) in comparison to the measured surface emissive power of 130 kW/m(2) whereas for d=3.4m these values are 180 and 250 kW/m(2). The predicted distance dependent irradiances agree well with the measured irradiances. PMID- 19185987 TI - Oxidative stress in prostate cancer. AB - As prostate cancer and aberrant changes in reactive oxygen species (ROS) become more common with aging, ROS signaling may play an important role in the development and progression of this malignancy. Increased ROS, otherwise known as oxidative stress, is a result of either increased ROS generation or a loss of antioxidant defense mechanisms. Oxidative stress is associated with several pathological conditions including inflammation and infection. ROS are products of normal cellular metabolism and play vital roles in stimulation of signaling pathways in response to changing intra- and extracellular environmental conditions. Chronic increases in ROS over time are known to induce somatic mutations and neoplastic transformation. In this review we summarize the causes for increased ROS generation and its potential role in etiology and progression of prostate cancer. PMID- 19185990 TI - Highly selective potentiometric determination of mercury(II) ions using 1-furan-2 yl-4-(4-nitrophenyl)-2-phenyl-5H-imidazole-3-oxide based membrane electrodes. AB - The electrode characteristics and selectivities of PVC-based mercury(II) selective coated graphite electrode (CGE) and polymeric membrane electrode (PME) incorporating the recently synthesized 1-furan-2-yl-4-(4-nitrophenyl)-2-phenyl-5H imidazole-3-oxide are reported here. The electrodes exhibit Nernstian slope for mercury(II) ions over wide concentration ranges, i.e. 1.0 x 10(-1)M to 1.0 x 10( 6)M (with CGE) and 1.0 x 10(-1)M to 1.0 x 10(-5)M (with PME). The lower detection limits shown by CGE and PME are 8.91 x 10(-7)M and 6.30 x 10(-6)M, respectively, in the pH range of 1.0-4.0. From the comparative study of these electrodes, CGE has been found to be better than PME in terms of lower detection limit and better selectivity for mercury(II) ions with comparatively less interference from silver(I) ions. The proposed electrodes can be successfully used as an indicator electrode for potentiometric titration of mercury with potassium dichromate. The electrodes have been successfully applied for estimation of mercury content in synthetic water samples, insecticide (parad tablet) and dental amalgam. PMID- 19185991 TI - Degradation of gas-phase trichloroethylene over thin-film TiO2 photocatalyst in multi-modules reactor. AB - The present paper examined the photocatalytic degradation (PCD) of gas-phase trichloroethylene (TCE) over thin-film TiO2. A large-scale treatment of TCE was carried out using scale-up continuous flow photo-reactor in which nine reactors were arranged in parallel and series. The parallel or serial arrangement is a significant factor to determine the special arrangement of whole reactor module as well as to compact the multi-modules in a continuous flow reactor. The conversion of TCE according to the space time was nearly same for parallel and serial connection of the reactors. PMID- 19185992 TI - Ecotoxicity tests using the green algae Chlorella vulgaris--a useful tool in hazardous effluents management. AB - The treatment efficiency of laboratory wastewaters was evaluated and ecotoxicity tests with Chlorella vulgaris were performed on them to assess the safety of their environmental discharge. For chemical oxygen demand wastewaters, chromium (VI), mercury (II) and silver were efficiently removed by chemical treatments. A reduction of ecotoxicity was achieved; nevertheless, an EC50 (effective concentration that causes a 50% inhibition in the algae growth) of 1.5% (v/v) indicated still high level of ecotoxicity. For chloride determination wastewaters, an efficient reduction of chromium and silver was achieved after treatment. Regarding the reduction of ecotoxicity observed, EC50 increased from 0.059% to 0.5%, only a 0.02% concentration in the aquatic environment would guarantee no effects. Wastewaters containing phenanthroline/iron (II) complex were treated by chemical oxidation. Treatment was satisfactory concerning chemical parameters, although an increase in ecotoxicity was observed (EC50 reduced from 0.31% to 0.21%). The wastes from the kinetic study of persulphate and iodide reaction were treated with sodium bisulphite until colour was removed. Although they did not reveal significant ecotoxicity, only over 1% of the untreated waste produced observable effects over algae. Therefore, ecotoxicity tests could be considered a useful tool not only in laboratory effluents treatment, as shown, but also in hazardous wastewaters management. PMID- 19185993 TI - Application of zeolitic material synthesized from thermally treated sediment to the removal of trivalent chromium from wastewater. AB - Zeolitic materials were synthesized from thermally treated sediment by alkali treatment using different NaOH/sediment ratios. Characterization of the materials was done by XRD, FTIR, cation exchange capacity and specific surface area. Use of high NaOH/sediment ratio favored the formation of zeolite. The potential value of the zeolitic materials for the retention of trivalent chromium from water was examined. The maximum of Cr(III) sorption by the zeolitic materials, determined by a repeated batch equilibration method, ranged from 38.9 to 75.8 mg/g which was much greater than that of the thermally treated sediment (6.3 mg/g). No release of sorbed Cr(III) by 1.0M MgCl(2) at pH 7 was observed but Cr(III) desorption by ionic electrolyte increased with decreasing pH. The zeolitic materials could completely remove Cr(III) from wastewater even in the presence of Na(+) and Ca(2+) with high concentrations with a dose above 2.5 g/L. The pH-dependent desorption behavior and the high selectivity of zeolitic material for Cr(III) were explained by sorption at surface hydroxyl sites and formation of surface precipitates. PMID- 19185994 TI - Study of photo-assisted electrochemical degradation of carbaryl at dimensionally stable anodes (DSA). AB - This paper presents the results concerning the degradation of the pesticide carbaryl comparing two methods: electrochemical (EC) and photo-assisted electrochemical (PAEC). The experimental variables of applied current density, electrolyte flow-rate and initial carbaryl concentration were investigated. The results demonstrate that the electrochemical degradation of carbaryl was greatly enhanced when simultaneous UV light was applied. The greatest difference between the PAEC and EC method was apparent when lower current densities were applied. The extent of COD removal was much enhanced for the combined method, independent of the applied current density. It should be noted that the complete removal of carbaryl was achieved with out the need to add NaCl to the reaction mixture, avoiding the risk of chlorinated organic species formation. PMID- 19185995 TI - Comparative study of the eliminating of waste gas containing toluene in twin biotrickling filters packed with molecular sieve and polyurethane foam. AB - Two kinds of packing materials, molecular sieve (MS) and polyurethane foam (PUF), were loaded into two identical biotrickling filters respectively to compare the microbial removal efficiency of waste gas containing toluene by seeding with same bacteria. The affecting parameters of the removal performance, such as gas flow rates, inlet toluene concentrations, periods of starvation, were investigated in detail in biotrickling filters. The results demonstrated that both of the packing materials exhibited high toluene degradation efficiency when the gas flow rates ranged from 100 L h(-1) to 600 L h(-1). For MS, the total maximum removal efficiency (RE) of toluene maintained 100% when the gas flow rates increased from 100 L h(-1) to 200 L h(-1) accompanied with the decrease of empty bed residence time (EBRT) from 266s to 133s. However, as for PUF, merely 97.64% RE was obtained at the gas flow rate of 100 L h(-1) and the EBRT of 266s. With further increasing the gas flow rates (to 600 L h(-1)) and decreasing the EBRTs (to 44s), both the total REs of toluene for MS and PUF decreased to 70.68% and 63.18%, respectively. When varying the inlet toluene concentrations, the REs for MS are able to maintain nearly 100% at the inlet concentration of 9.19 mg L(-1) or below, and with the maximum elimination capacity (EC) of 373.24 gm(-3)h(-1) (RE=100%) at the inlet concentration of 9.19 mg L(-1). Contrastively, the maximum EC of PUF was only 119.41 gm(-3)h(-1) (RE=56.66%) at the inlet concentration of 5.19 mg L(-1). As illustrated by different starvation period (2, 10 and 60 days), MS possessed shorter recovery time (9h for 2 days, 17 h for 10 days and 324 h for 60 days starvation, respectively) than PUF (14 h for 2 days, 24h for 10 days and 324 h for 60 days starvation, respectively). Based on its higher removal capacity of toluene and shorter recovery time, MS would be a better choice than PUF for packing material used for biotrickling filter. PMID- 19185996 TI - Biodegradation kinetics of picric acid by Rhodococcus sp.NJUST16 in batch reactors. AB - Biological degradation of 2,4,6-trinitrophenol (TNP) by Rhodococcus sp.NJUST16 in mineral salt medium was investigated in shake-flask experiments at pH of 7.0 and 30 degrees C, over a wide range of initial TNP concentration (20-800 mgl(-1)). The TNP was observed to be the inhibitory compound. For the studied concentration range, Haldane's model could be fitted to the growth kinetics data well with the kinetic constants mu(max)=0.2362 h(-1), K(s)=9.9131 mgl(-1) and K(i)=362.7411 mgl(-1). Further, the variation of observed yield coefficient Y with initial TNP concentration and the decay coefficient were investigated. It is our view that the above information would be useful for modeling and designing the units treating TNP-containing wastewaters. PMID- 19185997 TI - Non-isothermal thermal decomposition reaction kinetics of 2-nitroimino-5-nitro hexahydro-1,3,5-triazine (NNHT). AB - The thermal behavior and decomposition reaction kinetics of 2-nitroimino-5-nitro hexahydro-1,3,5-triazine (NNHT) were investigated by TG-DTG and DSC under atmospheric pressure and flowing nitrogen gas conditions. The results show that the thermal decomposition process of NNHT has two mass loss stages. The exothermic decomposition reaction mechanism obeys chemical reaction rule. The kinetic parameters of the reaction are E(a)=131.77 kJ mol(-1), lg(A/s(-1))=12.56, respectively. The kinetic equation can be expressed as: dalpha/dt = 10(12.86)(1 alpha)(3/2)3(-1.5849 x 10(4)/T)). The critical temperature of thermal explosion of NNHT obtained from the peak temperature (T(p)) is T(bp)=467.22K. The entropy of activation (DeltaS( not equal)), enthalpy of activation (DeltaH( not equal)), and free energy of activation (DeltaG( not equal)) of the reaction are -7.978 J mol(-1)K(-1), 127.99 kJ mol(-1) and 131.62 kJ mol(-1), respectively. PMID- 19185998 TI - Enhanced treatment of waste frying oil in an activated sludge system by addition of crude rhamnolipid solution. AB - The presence of high-strength oil and grease (O&G) in wastewater poses serious challenges for environment. Addition of surfactant into the activated sludge bioreactor is feasible in reducing high concentrations of O&G via enhancing its bioavailability. In this paper, an aqueous biosurfactant solution of rhamnolipid as a cell-free culture broth of Pseudomonas aeruginosa zju.um1 was added into a batch of aerobic activated sludge system for treatment of the waste frying oil. This treatment was conducted on both bench and pilot-scales, whereas the removal efficiency of frying oil was determined by analyzing the residue concentration of O&G and chemical oxygen demand (COD). In the presence of varying concentrations of rhamnolipid from 22.5 mg/L to 90 mg/L, aerobic treatment for 30 h was enough to remove over 93% of O&G while this biodegradability was only 10% in the control system with the absence of rhamnolipids. The equivalent biodegradability was similarly obtained on COD under addition of rhamnolipid. Compared with bench studies, a higher treatment efficiency with the presence of rhamnolipids was achieved on a pilot-scale of activated sludge system, in which a short time of 12h was required for removing approximately 95% of O&G while the control treatment attained a low efficiency of 17%. Finally, foaming and biodegradability of rhamnolipids in activated sludge system were further examined in the whole treatment process. It seems that the addition of rhamnolipid-containing culture broth showed great potential for treatment of oily wastewater by activated sludge. PMID- 19185999 TI - Chemical modification of carbonized wheat and barley straw using HNO3 and the adsorption of Cr(III). AB - The effects of oxidation using HNO(3) on the properties of the carbonized wheat and barley straw were investigated by measuring different properties such as specific surface area, PZC, total surface acidic groups as well as FTIR and TG DTA. A small decrease in the specific surface area due to pore blockage was observed after oxidation. After oxidation, the acidity was increased considerably and the point of zero charge shifted from approximately pH 9 to pH 2 in both types of carbon. By the oxidation of the carbon with nitric acid, carboxylic groups were produced as shown by absorption peaks at 1750 cm(-1) in the FTIR spectra. Boehm titration results showed that the number of carboxyl groups increased by approximately 11-fold after oxidation. The oxidized carbon samples were compared with the unoxidized original carbon samples for Cr(III) adsorption. It was observed that the oxidized carbons exhibit high adsorption efficiencies for Cr(III) ions compared to the original carbonized straws and this can be correlated to the decrease in PZC which is mainly due to the increase in carboxylic functional groups in the oxidized carbonized straws and these are responsible for their cation ion exchange phenomenon. PMID- 19186000 TI - Study of catalytic decomposition of formaldehyde on Pt/TiO2 alumite catalyst at ambient temperature. AB - Formaldehyde (HCHO) emitted from buildings, furnishing materials and consumer products is one of the most dominant volatile organic compounds (VOCs) in an indoor environment. In this work, a Pt/TiO(2)/Al(2)O(3) catalyst was prepared on an anodic alumite plate and was employed in the catalytic decomposition of formaldehyde at ambient temperature. Firstly, TiO(2) was deposited on the anodic alumite plate with electro-deposition technology. Then, platinum was supported on the anodic alumite plate with wet impregnation method. The developed catalyst exhibits good activity towards the decomposition of HCHO at ambient temperature. TPR (temperature programmed reduction) and TPD (temperature programmed desorption) analysis results indicate that oxygen adsorbed on the Pt/TiO(2)/Al(2)O(3) catalyst can be activated and generated to O:Pt(surface) species quickly at ambient temperature. Hence, the developed catalyst experiences the high activity towards the catalytic decomposition of formaldehyde at ambient temperature. Moreover, in accordance with the process requirements, the developed catalyst can be formed into various shapes such as a mesh, plate, fin, serrate etc., because aluminum can be formed into any shapes. The serrate type catalyst was prepared in this work and it also exhibits fine activity towards the decomposition of HCHO. PMID- 19186001 TI - Why do some patients keep hurting their back? Evidence of ongoing back muscle dysfunction during remission from recurrent back pain. AB - Approximately thirty-four percent of people who experience acute low back pain (LBP) will have recurrent episodes. It remains unclear why some people experience recurrences and others do not, but one possible cause is a loss of normal control of the back muscles. We investigated whether the control of the short and long fibres of the deep back muscles was different in people with recurrent unilateral LBP from healthy participants. Recurrent unilateral LBP patients, who were symptom free during testing, and a group of healthy volunteers, participated. Intramuscular and surface electrodes recorded the electromyographic activity (EMG) of the short and long fibres of the lumbar multifidus and the shoulder muscle, deltoid, during a postural perturbation associated with a rapid arm movement. EMG onsets of the short and long fibres, relative to that of deltoid, were compared between groups, muscles, and sides. In association with a postural perturbation, short fibre EMG onset occurred later in participants with recurrent unilateral LBP than in healthy participants (p=0.022). The short fibres were active earlier than long fibres on both sides in the healthy participants (p<0.001) and on the non-painful side in the LBP group (p=0.045), but not on the previously painful side in the LBP group. Activity of deep back muscles is different in people with a recurrent unilateral LBP, despite the resolution of symptoms. Because deep back muscle activity is critical for normal spinal control, the current results provide the first evidence of a candidate mechanism for recurrent episodes. PMID- 19186002 TI - The analgesic actions of centrally administered celecoxib are mediated by endogenous opioids. AB - Celecoxib is a selective inhibitor of cyclooxygenase-2 (COX-2) and blocks prostaglandin (PG) biosynthesis associated with inflammatory conditions. In a model of peripherally induced inflammatory pain in rats, celecoxib, given systemically, induced a state of hypoalgesia where the nociceptive threshold was raised above basal values, an effect not observed after treatment with non selective inhibitors of COX (indomethacin, piroxicam). Here, we have assessed the possibility that these atypical effects of celecoxib could be mediated by action at a site in the CNS. Inflammation and hyperalgesia were induced in one hind paw of rats by intraplantar injection of carrageenan (250microg). Nociceptive thresholds to mechanical stimulation were measured in the inflammed and contralateral paws for 6h after carrageenan injection. Celecoxib, SC236 (selective COX-2 inhibitors), indomethacin (non-selective COX inhibitor), SC560 (selective COX-1 inhibitor) or morphine were given by i.c.v. injection, 30 min before carrageenan. Celecoxib, SC236 or morphine-induced hypoalgesia whereas, after indomethacin or SC 560, the nociceptive threshold only returned to basal values. Naltrexone, also given i.c.v., reversed the hypoalgesia after celecoxib or morphine. Bestatin, an inhibitor of metabolism of endogenous opioid peptides, given i.c.v., potentiated the analgesic effects of a low dose of celecoxib. Taken together, these data indicate that celecoxib could act centrally after systemic administration to produce its characteristic profile of analgesia in this model of peripheral inflammatory pain. Moreover, this atypical analgesia appeared to be mediated by endogenous opioids rather than by inhibition of PG biosynthesis. PMID- 19186003 TI - The relationship between high catastrophizing children's facial display of pain and parental judgment of their child's pain. AB - The present study investigated the effect of the child's pain catastrophizing and self-reported pain upon the child's facial expression of pain and parental inferences of their child's pain. School children (n=62) experienced pain by taking part in a cold water procedure. Analyses revealed that more intense pain was associated with higher levels of facial pain expression in children who reported a low frequency of catastrophizing. In children with high pain catastrophizing, this relationship was not significant. A similar pattern was obtained for the pain inferences by the parent: pain intensity as reported by the child was positively related to pain inferences by the parent in children who reported a low frequency of catastrophizing, but such relationship was not significant for children with high catastrophizers. Further analyses revealed that when pain intensity was low, parents of high catastrophizing children judged the pain of their child to be higher than parents of low catastrophizing children. The implications of the findings are discussed in terms of the importance of assessing different dimensions of pain encoded in expression, different types of pain expression, and its differential effects upon others. PMID- 19186004 TI - Tell me why... The influence of self-involvement on short term smoking behaviour. AB - Previous studies found that information is more persuasive when self-generated (high self-involvement), rather than when simply read or heard (low self involvement). In two studies, we investigated whether differences in self involvement concerning smoking issues would influence immediate smoking behaviour. As predicted, results indicate that participants who developed their own arguments against smoking waited longer before lighting up a cigarette than those who read arguments against smoking that were developed by other participants (Study 1). Further, participants who additionally generated their own arguments were less likely to smoke within 30 min than those who read prepared arguments against smoking (Study 2). In sum, our studies illustrate that personal involvement in generating anti-smoking arguments can reduce short term smoking behaviour. PMID- 19186005 TI - Epigenetic alteration of p16INK4a gene in dedifferentiation of liposarcoma. AB - The atypical lipomatous tumor (ALT)/well-differentiated liposarcoma (WDLPS) is a locally aggressive subtype of liposarcoma unless dedifferentiation occurs. The mechanism driving this progression is not clear. Loss of p16 is believed to be an early and critical event in tumor progression. Gene silencing by methylation of p16INK4a gene promoter has been reported in several soft tissue sarcomas. The aim of this study is to study the role of p16INK4a gene promoter methylation and p16 expression in tumor progression (dedifferentiation) and recurrence of ALT/WDLPS. Four cases of dedifferentiated liposarcomas (DDLPS) and three cases of recurrent well-differentiated liposarcomas (WDLPS) were collected, and methylation status of p16INK4a gene promoter was analyzed using methylation-specific PCR (MSP) on DNA extracted from paraffin blocks. p16 expression was examined by immunohistochemistry on the same blocks. Methylation of p16INK4a gene promoter was seen in the dedifferentiated (DD) components only, in two out of four (2/4, 50%) DDLPS. The other two DDLPS and three recurrent WDLPS were not methylated. Both WD and DD components in all four DDLPS cases showed strong nuclear p16 expression. All three recurrent WDLPS showed positive p16 expression with similar intensity between primary and recurrent tumors. Even though linear correlation between p16 promoter hypermethylation and p16 protein expression was not present, there appears to be a role for p16INK4a gene promoter hypermethylation in DDLPS and not in recurrent WDLPS. PMID- 19186006 TI - Prevalence of infectious pathogens in Crohn's disease. AB - The importance of infectious pathogens in Crohn's disease (CD) is still under debate. Therefore, we examined a panel of potential viral and bacterial pathogens in a large series of CD patients and controls. Archival tissue from 76 patients, 56 with CD and 20 control patients, with normal colon mucosa (n=10) and non steroid anti-inflammatory drug (NSAID)-induced colitis (n=10) were examined using PCR-based detection methods for human cytomegalovirus (CMV), Epstein-Barr virus (EBV), herpes simplex virus 1, 2 (HSV1,2), adenovirus (AD), varicella-zoster virus (VZV), human herpes virus 6 (HHV6), human herpes virus 8 (HHV8), Mycobacterium tuberculosis complex (Mtbc), atypical mycobacteria (nM/MG1), including Mycobacterium avium (subspecies paratuberculosis, MAP), Stenotrophomonas maltophilia (Sm), and Yersinia enterocolitica (Ye). In CD patients, positive PCR results were achieved in 19 cases (34%). Sm was most frequent in 10 of 56 cases (17.9%) followed by EBV (6/56, 10.7%), nM/MG1 (4/56, 7.1%), including MAP, HHV6, and CMV (2/56, 3.6%), and finally Mtbc and AD (1/56, 1.8%). The control patients showed positive PCR results in 12 patients (12/20, 60%), nine of them with only weak signals, suggesting a persistent infection. In addition, we compared typical pathomorphological features of CD patients with the PCR results and found a significant correlation between EBV infection and mural abscesses (P=0.014). Our data demonstrate that several potential pathogens can be detected in a sizeable fraction of specimens from patients with CD, but also in control patients, suggesting that the analyzed infectious pathogens may be associated with the disease, but do not represent an obligatory cause. PMID- 19186007 TI - Differential expression of survivin and its splice variants, survivin-DeltaEx3 and survivin-2B, in bladder cancer. AB - BACKGROUND: Survivin, an inhibitor of apoptosis protein (IAP), has been regarded as a valuable tumor marker for diagnosis and prognosis of bladder cancer. Recently, three splice variants of the gene with different anti-apoptotic activities have been reported. However, there is no data on the expression and potential causative roles of these transcripts in bladder cancer. Here, we have investigated the expression pattern of survivin and two of its splice variants, survivin-DeltaEx3 and survivin-2B, in malignant versus non-malignant bladder tissues. METHODS: We used semi-quantitative RT-PCR analysis to examine the expression of survivin variants in 30 transitional cell carcinoma, 19 matched non tumor, and 9 apparently healthy control tissue samples of the bladder. DNA sequencing was used to confirm the identity of amplified fragments. RESULTS: For all examined samples, survivin was the predominant transcript, being present in 83% of tumor and 25% of non-tumor bladder tissue samples, and survivin-2B was the least detected isoform. The expression levels of survivin and survivin-DeltaEx3 was significantly higher in neoplastic compared to non-neoplastic tissues (P<0.001) and both the sensitivity and specificity of survivin were superior to survivin-DeltaEx3 (83% and 75% for survivin and 76% and 64% for survivin DeltaEx3, respectively). Also, the expression levels of survivin and survivin DeltaEx3 showed a significant correlation (P=0.02) with tumor invasiveness (pT1/pTa). CONCLUSION: Our data revealed for the first time a differential expression pattern of survivin splice variants in bladder tissues, which potentially could have a practical usefulness in diagnosis and/or therapy of the tumor. PMID- 19186009 TI - Geographical dissemination of Leptospira interrogans serovar Pomona during seasonal migration of California sea lions. AB - Leptospirosis is one of the most widespread bacterial zoonoses in the world and affects most mammalian species. Although leptospirosis is well documented and characterized in terrestrial species, less information is available regarding the distribution and impact of leptospirosis in marine mammals. Additionally, the role of animal migrations on the geographical spread of leptospirosis has not been reported. Periodic epizootic outbreaks of acute leptospirosis among California sea lions (Zalophus californianus) have been reported since 1971. In this study, we collected samples from California sea lions stranded along the Pacific coast of North America during the most recent epidemic in 2004, and maintained leptospirosis surveillance of the California sea lion population along the California coast through 2007. Several isolates of Leptospira interrogans serovar Pomona were obtained from kidney and urine samples collected during this study, a finding consistent with serological evidence that California sea lions are persistently exposed to this leptospiral serovar. Combined, these data support a model whereby California sea lions are maintenance hosts for L. interrogans serovar Pomona, yet periodically undergo outbreaks of acute infection. During the 2004 outbreak, the incidence of new leptospirosis cases among California sea lions coincided with the seasonal movement of male sea lions from rookeries along the coast of central and southern California north as far as British Columbia. These data show that seasonal animal movement contributes to the distribution of leptospirosis across a large geographical region. PMID- 19186008 TI - Differential effects of cholesterol and phytosterols on cell proliferation, apoptosis and expression of a prostate specific gene in prostate cancer cell lines. AB - BACKGROUND: The purpose of our study was to show the apoptotic and anti proliferative effects of phytosterols as distinct from cholesterol effects on prostate cancer cell lines, and also their differential expression of caveolin-1, and a prostate specific gene, PCGEM1. METHODS: PC-3 and DU145 cells were treated with sterols (cholesterol and phytosterols) for 48h, followed by trypan blue dye exclusion measurement of cytotoxicity and MTT cell proliferation assays, respectively. Cell cycle analysis was carried out microscopically, and by propidium iodide uptake using flow cytometry. Sterol induction of oncogenic gene expression was evaluated by RT-PCR. Apoptotic cells were identified by immunocytochemistry using DNA fragmentation method, and by annexin V adhesion using flow cytometry. RESULTS: Physiological doses (16microM) of these sterols were not cytotoxic in these cells. Cholesterol-enrichment promoted mitosis (54 and 61% by microscopy; 40.8 and 34.08% by FACS analysis in PC-3 and DU145, respectively) and cell growth (P<0.05), while phytosterols suppressed mitosis (29 and 35% by microscopy; 27.71 and 17.37% by FACS analysis in PC-3 and DU145, respectively), and significantly induced tumor-suppression (P<0.05) and apoptosis. We demonstrated for the first time that cholesterols upregulated the expression of PCGEM1 even in androgen-insensitive prostate cancer cell lines. Phytosterols reversed this effect, while upregulating the expression of caveolin 1, a known mediator of androgen-dependent proto-oncogene signals that presumably control growth and anti-apoptosis. CONCLUSIONS: Phytosterol inhibition of PCGEM1 and cell growth and the overexpression of caveolin-1, suggests that poor disease prognosis anchors on the ability of caveolin-1 to regulate downstream oncogene(s) and apoptosis genes. Sterol intake may contribute to the disparity in incidence of prostate cancer, and elucidation of the mechanism for modulation of growth and apoptosis signaling may reveal potential targets for cancer prevention and/or chemotherapeutic intervention. Sterol regulation of PCGEM1 expression suggests its potential as biomarker for prediction of neoplasms that would be responsive to chemoprevention by phytosterols. PMID- 19186010 TI - Respiratory disease in calves: microbiological investigations on trans-tracheally aspirated bronchoalveolar fluid and acute phase protein response. AB - Trans-tracheal aspirations from 56 apparently healthy calves and 34 calves with clinical signs of pneumonia were collected in six different herds during September and November 2002. The 90 samples were cultivated and investigated by PCR tests targeting the species Histophilus somni, Mannheimia haemolytica, Pasteurella multocida, Mycoplasma bovis, Mycoplasma dispar, and Mycoplasma bovirhinis. A PCR test amplifying the lktC-artJ intergenic region was evaluated and shown to be specific for the two species M. haemolytica and Mannheimia glucosida. All 90 aspirations were also analyzed for bovine respiratory syncytial virus (BRSV), parainfluenza-3 virus, and bovine corona virus by antigen ELISA. Surprisingly, 63% of the apparently healthy calves harbored potentially pathogenic bacteria in the lower respiratory tract, 60% of these samples contained either pure cultures or many pathogenic bacteria in mixed culture. Among diseased calves, all samples showed growth of pathogenic bacteria in the lower respiratory tract. All of these were classified as pure culture or many pathogenic bacteria in mixed culture. A higher percentage of the samples were positive for all bacterial species in the group of diseased animals compared to the clinically healthy animals, however this difference was only significant for M. dispar and M. bovirhinis. M. bovis was not detected in any of the samples. BRSV was detected in diseased calves in two herds but not in the clinically healthy animals. Among the diseased calves in these two herds a significant increase in haptoglobin and serum amyloid A levels was observed compared to the healthy calves. The results indicate that haptoglobin might be the best choice for detecting disease under field conditions. For H. somni and M. haemolytica, a higher percentage of the samples were found positive by PCR than by cultivation, whereas the opposite result was found for P. multocida. Detection of P. multocida by PCR or cultivation was found to be significantly associated with the disease status of the calves. For H. somni a similar association with disease status was only observed for cultivation and not for PCR. PMID- 19186011 TI - Evaluation of the immunogenicity of recombinant stress-associated proteins during Mycobacterium avium subsp. paratuberculosis infection: Implications for pathogenesis and diagnosis. AB - The aim of this study was to assess the immunogenicity of recombinant stress associated proteins of Mycobacterium avium subsp. paratuberculosis in sheep infected with the organism compared to control sheep. Five proteins - MAP2411, ClpP, Ppa, MAP0593c and GreA - which were identified previously in in vitro stress or dormancy responses of M. paratuberculosis to hypoxia, nutrient starvation and heat, were cloned, expressed and purified as His-tag recombinant proteins from the pET-15b vector in a BL21(DE3)pLysS strain of E. coli. The immunogenicity of MAP2411 did not differ between infected and control sheep. However, the serological reactivity of the other recombinant antigens, and combinations of them, varied according to the histological stage of paratuberculosis. Interestingly, the sera from some animals with paucibacillary lesions, which were not immunoreactive in a commercial paratuberculosis ELISA that was based on non-defined native antigens, recognised the recombinant antigens. We infer from their differential immunogenicity in infected and control sheep that four of the stress-associated proteins were expressed by M. paratuberculosis in vivo. These data provide fundamental information on host mycobacterial interactions and have conceptual implications for the development of future diagnostic tests for early immune responses in animals infected with mycobacteria. PMID- 19186012 TI - Patent ductus arteriosus: perspectives on a 'persistent' problem. PMID- 19186013 TI - Gender-related effects of prenatal administration of estrogen and progesterone receptor antagonists on VEGF and surfactant-proteins and on alveolarisation in the developing piglet lung. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) is essential for embryonic lung development and has been shown to be regulated by estradiol (E2) and progesterone (P). AIM: To investigate the effects of prenatal E2 and P withdrawal by specific receptor antagonists on the mRNA expression of VEGF, surfactant proteins (SP-B and SP-C) and on alveolarisation in lung tissue of male and female pig fetuses. METHODS: Fetuses from 10 sows were randomized to receive either both an intramuscular injection of the E2 receptor blocker ICI 182.780 and the P receptor blocker RTI 3021-022 (ICI+RTI, n=5) or a placebo injection (n=5) at 90 days of gestation (DOG, 115=term). After delivery by cesarean section on 114 DOG, tissue of the left lingula of the piglet's lung (28 placebo, 26 ICI+RTI) was obtained to determine the mRNA expression of VEGF, SP-B and SP-C. Lungs from 15 placebo and 14 ICI+RTI group piglets were removed and alveolar counts performed. RESULTS: The ICI+RTI group showed significantly lower SP-C mRNA expression and alveolar counts compared to the placebo group (p=0.04 and 0.03, respectively). Diminished alveolarisation in the ICI+RTI group was mainly due to the reduction of alveolar counts in male piglets (p=0.02). Within the placebo group VEGF and SP B mRNA expression in male piglets were significantly lower compared to female piglets (p=0.01 and 0.004, respectively). ICI+RTI treatment abolished this gender related difference. CONCLUSION: Estradiol and P antagonism affected gender related differences of key proteins for pulmonary function and development and especially in males was associated with diminished alveolarisation. PMID- 19186014 TI - Sexuality during the climacteric period. AB - BACKGROUND: Cultural, social, physiological and psychological factors may alter the course of sexual function in climacteric women. OBJECTIVE: The objective of the present literature review is to survey the prevalence of sexual dysfunctions in the climacteric and to establish the association between the organic and psychic changes that occur during this phase and sexual dysfunction. We also discuss potential treatments. METHODS: We evaluated the data available in PubMed (1982-2008). For each original article, two reviewers analyzed the data independently and considered a study to be of high quality if it had all three of the following characteristics: prospective design, valid data and adequate sample size. Both reviewers extracted data from each of the 99 studies selected: 34 cross-sectional studies, 25 cohort studies, 9 trials, 31 reviews related to sexuality in pre- and post-menopausal women. RESULTS: Sexual dysfunction among climacteric women is widespread and is associated with bio-psychosocial factors. However, there is not enough evidence to correlate sexual dysfunction with a decrease in estrogen levels and biological aging. A strong association exists between climacteric genital symptoms and coital pain. There is, however, sufficient evidence demonstrating the benefits of local estrogen therapy for patients with genital symptoms. CONCLUSION: A significant decline in sexual function occurs in climacteric women, although it is still unclear whether this is associated with the known decrease in estrogen levels or with aging, or both. Relational factors may interfere with sexual function during this phase. The climacteric genital symptoms improve with estrogen replacement therapy, and positively influence sexual function. Further studies are needed to establish the actual impact of the decrease in estrogen levels and of aging on the sex life of climacteric women. PMID- 19186015 TI - Vibrio gallaecicus sp. nov. isolated from cultured clams in north-western Spain. AB - A group of three motile facultative anaerobic marine bacteria were isolated from cultured Manila clams (Ruditapes philippinarum) in Galicia, north-western Spain. The strains were characterized phenotypically and genotypically. Phylogenetic analysis of the 16S rRNA gene and four housekeeping genes, RNA polymerase alpha chain (rpoA), RecA protein (recA), the alpha-subunit of bacterial ATP synthase (atpA) and the uridine monophosphate (UMP) kinase (pyrH), indicated that these strains were closely related to the Vibrio splendidus clade. The amplified fragment length polymorphism (AFLP) fingerprints, DNA-DNA hybridizations and phylogenies of the housekeeping and 16S rRNA gene sequences showed that the three strains represented a different species from all currently described vibrios. The new species could be differentiated from its nearest neighbours on the basis of several phenotypic features. The three strains are therefore a novel species within the genus Vibrio, for which the name Vibrio gallaecicus is proposed, with the type strain being VB 8.9T(=CECT 7244T=LMG 24045T). PMID- 19186016 TI - Overcoming extractability hurdles of a 14C labeled taxane analogue milataxel and its metabolite from xenograft mouse tumor and brain tissues. AB - Taxane analogue milataxel have been shown to bound to proteins/tissues irreversibly. Extraction of the aforementioned bound drug and metabolite was proven to be difficult task. Nonetheless, an extraction method had to be developed to accurately determine drug concentration in tissues over time. This method would enable Taxolog, Inc. (Fairfield, NJ, USA) to accurately map the fate of drug in mice and it would also enable to better design drug dosing scheme for its maximum efficiency. A productive extraction technique for milataxel (MAC-321, TL-139) in nude mice with various xenograft human tumors was developed by extracting analytes from tumors using a novel extraction procedure and analyzing samples by LC-MS. This extraction technique entails disrupting tissue cells with hexane followed by acidic methanol (MeOH), with the aid of a tissuemizer and sonic cell disrupter. An average extractability of 75% was achieved as confirmed by the recovery of 14C labeled milataxel, as compared to 4-48.5% extraction efficiency using solvents and/or combination of solvents such as acetonitrile (ACN), ethanol, ethyl acetate, MeOH/acetic acid in water, and chloroform/MeOH. This extraction technique allowed for quantitation of milataxel and its major metabolite s-lactate (M-10) from tumors and brain tissue samples using HPLC coupled with electro-spray ionization mass spectrometry (HPLC-ESI-MS). Ratios of M-10 metabolite to milataxel were determined to be approximately 3:1 and 2:1 in SKMES human lung carcinoma tumors and A-375 melanoma tumors, respectively, and declined in concentration over 20 days. However, levels of milataxel and M-10 were determined to be equal at 8h in HCT-15 human colon carcinoma tumors with M 10 levels dropping sharply over a 10-day period. PMID- 19186017 TI - The use of differential scanning calorimetry for the purity verification of pharmaceutical reference standards. AB - Reference standards are routinely used in pharmaceutical industry to determine strength, content, and the quality of drug products, active pharmaceutical ingredients (API), preservatives, antioxidants and excipients. Traditionally, chromatographic techniques such as High Performance Liquid Chromatography (HPLC) and Gas Chromatography (GC) in conjunction with other analytical techniques have been used to determine the purity and strength of a specific lot of a compound for the purpose of qualifying the lot to use as a reference standard. The assigned purity of the reference standard for a wide variety of compounds can be verified using an absolute method such as Differential Scanning Calorimetry (DSC). In this paper, purity of 16 reference standards was determined by DSC and the results were then compared to the purity values that were obtained using HPLC and other analytical techniques. The results indicate that the purity obtained from DSC analysis is comparable to the chromatographic purity for organic compounds that are at least 98% pure. Use of DSC for purity determination is not appropriate if a compound lacks sharp melting point, decomposes in the defined temperature range or exhibits other thermal event(s) which interfere with the melting point of the compound. The use of DSC as an alternative and or complementary method to verify the purity of a compound as part of the pharmaceutical reference standard certification process is discussed. PMID- 19186018 TI - Profiling and quantification of isoflavone-C-glycosides impurities in puerarin injection by liquid chromatography coupled to ESI-ion trap mass spectrometry. AB - An HPLC/DAD/MS(n) method was established for the qualitative and quantitative analysis of the impurities in puerarin injection (PI), a widely used drug in China. The analytical HPLC was performed on an Agela RP-C18 column using 0.1% aqueous formic acid (v:v) and methanol as mobile phase. A total of nine impurities were detected and eight of them were identified as isoflavone-C glycosides basing on their UV spectra and MS(n) spectra and comparing with the literature data. An HPLC method for the assay of two common impurities in the commercial PI samples, i.e., neopuerarin A and neopuerarin B, was then established. The validation of the method, including sensitivity, linearity, precision, accuracy, was carried out. The calibration curves showed good linearity of R(2)>0.9999 and LOQ (S/N=10) were less than 3.73 ng. The precision was evaluated by intra- and inter-day assays and R.S.D. values were less than 0.94%. The average recovery rates were 97.0% and 99.5%, respectively, with R.S.D. less than 1.38%. The contents of neopuerarin A and neopuerarin B in various commercial brands of PI samples varied over the range of 0.30-1.16% and 0.42 1.66%, respectively. This is the first report on the impurities in PI. PMID- 19186019 TI - Volatile composition of Catharanthus roseus (L.) G. Don using solid-phase microextraction and gas chromatography/mass spectrometry. AB - A total of 88 volatile and semi-volatile components were formally or tentatively identified in flowers, leaves and stems of Catharanthus roseus (L.) G. Don (cv. Little Bright Eye), by headspace solid-phase microextraction (HS-SPME) and by dichloromethane extraction, combined with gas chromatography-mass spectrometry (GC-MS). These include some diterpenic compounds (manool and manoyl oxides), a sesquiterpen (alpha-bisabolol), and some pyridine, pyrazine, indol and carotenoid derivatives. Applying multivariate analysis (principal component analysis and agglomerative hierarchic cluster analysis) to the HS-SPME-GC-MS data, it was possible to characterize each part of the vegetal material using a relative small number of compounds. Hence, flowers were richer in terpenic molecules (including limonene), alpha-bisabolol, methyljasmonate, cis-jasmone, 2-phenylethanol, phenylacetaldehyde, trans-2-octenal, benzylic alcohol and 2-isobutyl-3 methoxypyrazine. Leaves can be characterized by the methyl and propyl esters of fatty acids, mono- and disaturated, trans-phytol, carotenoid derivative compounds, hydrofarnesylacetone, methylanthranilate, manool and epi-manool oxide, while stems have high levels of volatile aldehydes, such as hexanal, octanal, cis 2-nonenal, cis-2-decenal, cis, trans-2,6-nonadienal, trans, trans-2,4-decadienal and cis, trans-2,4-decadienal. Dichloromethane extraction allowed also the identification of some alkaloid-like compounds that were not detected by HS-SPME. PMID- 19186020 TI - Electrochemical DNA nano-biosensor for the study of spermidine-DNA interaction. AB - An electrochemical DNA nano-biosensor is prepared by immobilization of double stranded DNA (dsDNA) onto a mixed self-assembled monolayer (SAM) composed of azide- and hydroxyl-terminated thiols. The SAMs- and dsDNA-modified gold electrodes were characterized by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The interaction of spermidine with DNA was studied by differential pulse voltammetry at the DNA-modified electrode. The decrease in the guanine oxidation peak current was used to study the interaction. The binding constant (K), obtained by differential pulse voltammetry, was 1.85 x 10(5)M(-1). A linear dependence of the guanine peak currents was observed in the range of 1.6 70.4 microM spermidine, with a detection limit of 0.72 microM and r=0.994 by using differential pulse voltammetry. PMID- 19186021 TI - Determination of breviflavone A and B in Epimedium herbs with liquid chromatography-tandem mass spectrometry. AB - Two new types of minor flavonoids, breviflavone A and B, have been recently isolated and identified from Epimedium brevicornu in our previous research. Breviflavone B is a novel flavonoid with potent and specific estrogen receptor (ER) bioactivity. Its positional isomer, breviflavone A, is not ER active. Therefore, it is important to determine the two minor components, breviflavone A and B, in Epimedium herbs. In this report, a robust method for measurement of the two breviflavones in Epimedium ethanolic extracts has been developed by using liquid chromatography tandem mass spectrometry via selected-reaction monitoring (m/z 437-->m/z 367 for breviflavone A and m/z 437-->m/z 351 for breviflavone B) under negative electrospray ionization mode. This method has been successfully used to determine the two breviflavones in ethanolic herbal extracts of five major Epimedium species (E. brevicornu, E. koreanum, E. pubescens, E. sagittatum, and E. wushanese) from various sources. The contents of the two breviflavones range from 0.0181 to 0.1791% for breviflavone A and 0.0026 to 0.0252% for breviflavone B in the dried ethanolic extracts of those Epimedium herbal samples. PMID- 19186022 TI - [Benefits of ultrasound-guided axillary brachial plexus block in patients with arteriovenous fistulae]. PMID- 19186023 TI - [Patient-controlled analgesia. Pain and Locoregional Anesthesia Committee and the Standards Committee of the French Society of Anesthesia and Intensive Care]. PMID- 19186024 TI - [Ultrasound and regional anaesthesia]. AB - The use of ultrasound is the latest major evolution in regional anaesthesia. Review of available literature shows significant changes in clinical practice. Ultrasound guidance allows the visualization of anatomical variations or unsuspected intraneural injections, reduces the volume of local anaesthetic injections and confirms correct local anaesthetic distribution or catheter placement. No study has found a statistical difference in success rates and safety because all studies were underpowered. However, the ability to visualize an invasive procedure that has been performed blindly in the past is an undeniable progress in terms of safety. The necessity to be familiar with the machine and the learning curve can be repulsive. The aim of this article is to demystify ultrasound guidance by explaining the fundamentals of the clinical use of ultrasound. With the help of different chapters, the authors explain the different adjustments and possible artefacts and give easy solutions for the use of bedside ultrasound. Training is essential and can be performed on manikins or training phantom. For each region the main anatomical landmarks are explained. One must be familiar with several imaging techniques: short axis (transverse) or long axis (longitudinal) nerve imaging, in-plane or out-of-plane imaging and hydrolocalization. Viewing the needle's tip position during its progression remains the main safety endpoint. Therefore, electrical nerve stimulation and ultrasound guidance should be combined, especially for beginners, to confirm proximity to neural structures and to help in case of difficulty. Optimizing safety and clinical results must remain a key priority in regional anaesthesia. Finally, specific regulations concerning the transducers are described. Paediatric specificities are also mentioned. PMID- 19186025 TI - [Vision loss after spine surgery: a case report]. AB - We report the case of a 36-year-old man who underwent neurosurgery for a T9 spine fracture consecutive to a fall. The patient had complete postoperative blindness which did not totally recover during the hospital stay. Decreased visual acuity and postoperative vision loss are not uncommon in spine surgery. Such postoperative complications in spine surgery are severe. To avoid them, it is mandatory to identify the contributing factors and set up a preventive strategy. PMID- 19186026 TI - [Postoperative pain management using regional anaesthesia in the child. Pain and Locoregional Anesthesia Committee and the Standards Committee of the French Society of Anesthesia and Intensive Care]. PMID- 19186027 TI - [Morbidity and mortality of patients with preeclampsia or HELLP syndrome transferred in intensive care]. AB - CONTEXT: During pregnancy, the two leading causes of admission in intensive care are preeclampsia and obstetric haemorrhage. However, there are few studies about preeclamptic patients admitted in intensive care. Our purpose was to determine the outcome of pregnancies with preeclampsia and/or HELLP syndrome admitted in intensive care. METHODS: We performed a retrospective study between March 1996 and April 2005 in a level III maternity. 533 patients were managed with preeclampsia and/or HELLP syndrome during this period. We compared patients admitted in intensive care with patients who did not require admission in intensive care. RESULTS: Sixty six patients (12,4%) with preeclampsia and/or HELLP were admitted in intensive care. Severe HELLP syndrome, eclampsia, neurological troubles and acute pulmonary oedema were the four leading causes of admission. The mean duration of admission was 3, 2+/-2,9 days. Mean age of the patients (28, 2+/-5,8 vs. 29,0+/-5,8 years, NS) and number of primiparous (71, 2% vs 66,6%, NS) were similar between the two groups. The mean gestational age of delivery was reduced when patients were needed admission in intensive care (29,8+/-3,9 weeks of gestation versus 32,5+/-4,4, p<0,001). 77, 3% of babies survived in the intensive care group compared with 90,4% in the other group (p<0,01). CONCLUSION: The short-term outcome of patients admitted in intensive care for preeclampsia or HELLP syndrome is generally good. However, neonatal morbidity and mortality remained important when women needed management in intensive care. PMID- 19186028 TI - Hypnosis for postradiation xerostomia in head and neck cancer patients: a pilot study. AB - Xerostomia, the sensation of dry mouth, affects almost all patients who undergo radiotherapy for cancer in the head and neck area. Current therapies for xerostomia are inadequate, and the condition negatively impacts the quality of life. This prospective observational pilot study aimed to evaluate whether hypnosis could improve salivation and decrease xerostomia. Twelve patients with xerostomia after radiotherapy for head and neck cancer were assessed for severity of xerostomia symptoms and sialometry. They then received a single hypnosis session with specific suggestions to increase salivation. The session was recorded on a compact disk (CD), and the participants were instructed to listen to it twice a day for one month. Sialometry was repeated immediately after hypnosis. Validated xerostomia questionnaires were completed at one, four, and 12 weeks after hypnosis. A substantial overall improvement was reported by eight patients at 12 weeks (66%). The saliva flow rate increased on sialometry in nine patients following hypnosis (75%). There was no correlation between the magnitude of changes in the measured saliva flow rate and changes in subjective measures (Spearman's correlation coefficient r=0.134). Symptomatic improvement significantly correlated with the number of times the patients listened to the hypnosis CD (r=0.714, P=0.009). No adverse events were reported. The data from this small observational trial suggest that hypnosis may be an effective treatment for xerostomia. Confirmation in a larger randomized and controlled investigation is warranted. PMID- 19186029 TI - Effects of SC58236, a selective COX-2 inhibitor, on epileptogenesis and spontaneous seizures in a rat model for temporal lobe epilepsy. AB - Inflammation is an important biological process that is activated after status epilepticus and could be implicated in the development of epilepsy. Here we tested whether an anti-inflammatory treatment with a selective cox-2 inhibitor (SC58236) could prevent the development of epilepsy or modify seizure activity during the chronic epileptic phase. SC58236 was orally administered (10mg/kg) during the latent period for 7 days, starting 4h after electrically induced SE. Seizures were monitored using EEG/video monitoring until 35 days after SE. Cell death and inflammation were investigated using immunocytochemistry (NeuN and Ox 42). Sprouting was studied using Timm's staining after 1 week and after 4-5 months when rats were chronic epileptic. SC58236 was also administered during 5 days in chronic epileptic rats. Hippocampal EEG seizures were continuously monitored before, during and after treatment. SC58236 effectively reduced PGE(2) production but did not modify seizure development or the extent of cell death or microglia activation in the hippocampus. SC58236 treatment in chronic epileptic rats did not show any significant change in seizure duration or frequency of daily seizures. The fact that cox-2 inhibition, which effectively reduced prostaglandin levels, did not modify epileptogenesis or chronic seizure activity suggests that this type of treatment (starting after SE) will not provide an effective anti-epileptogenic or anti-epileptic therapy. PMID- 19186030 TI - The efficacy of lamotrigine in clozapine-resistant schizophrenia: a systematic review and meta-analysis. AB - BACKGROUND: Clozapine is the drug of choice for patients with unsatisfactory response to routine antipsychotic treatment. Polypharmacy is widely used among patients having clozapine-resistant schizophrenia, although no solid evidence exists for any effective augmentation therapy for this patient population. We aimed to study the efficacy of lamotrigine in the treatment of clozapine resistant schizophrenia. METHOD: We conducted electronic searches of the Cochrane PsiTri database, the Website of metaRegister of Controlled Trials, including NIH ClinicalTrials.gov, and a clinical trial register by the manufacturer of lamotrigine (GlaxoSmithKline). All randomized placebo-controlled studies on patients receiving clozapine were included in the analysis. The primary outcome measure was a total score for symptoms of psychosis, and the secondary outcome measures were scores for positive and negative symptoms of psychosis. For continuous and binary data, standardized mean differences (SMD), and odds ratios (OR) and the number needed to treat (NNT) were calculated, respectively. RESULTS: Five trials with 10 to 24 weeks duration and total of 161 randomized clozapine patients were included in the meta-analysis. Lamotrigine was superior to placebo augmentation in both the primary outcome measure (SMD 0.57, 95%CI 0.25-0.89, p<0.001; OR 0.19, 95%CI 0.09-0.43, p<0.001; NNT 4, 95%CI 3-6) and secondary outcome measures (SMD 0.34, 95%CI 0.02-0.65 for positive symptoms, SMD 0.43, 95%CI 0.11-0.75 for negative symptoms). CONCLUSIONS: This meta-analysis suggests that lamotrigine augmentation may be an effective treatment for patients with clozapine-resistant schizophrenia. A substantial proportion of these most severely ill patients appeared to obtain clinically meaningful benefit from this combination treatment. PMID- 19186031 TI - Silencing the androgen receptor: new skills for antiandrogen oligonucleotide skin and hair therapy. PMID- 19186032 TI - Interaction of dopamine D1 with NMDA NR1 receptors in rat prefrontal cortex. AB - Despite the tremendous importance of D1 and NMDA receptors to cognition (working memory, executive functions) and synaptic plasticity in the prefrontal cortex (PFC), little is known about the molecular mechanisms underlying D1-NMDA receptors interactions in this brain area. Here, we show that D1 receptors and the NMDA receptor co-localize in single pyramidal neurons and interneurons in adult rat PFC. NR1 and NR2A expression are found in different neuronal types. Conversely, D1 receptors are predominantly localized in pyramidal-like cells and parvalbumin positive cells. NR1 co-immunoprecipitates with D1 receptor in adult medial PFC. In prefrontal primary cultures, NMDA does not affect the D1 receptor dependent-cAMP production. In contrast, activation of D1 receptor potentiates the NMDA mediated increase in cytosolic Ca2+, an effect that was blocked by a PKA inhibitor. We conclude that D1 receptor potentiates the NMDA-Ca2+ signal by a PKA dependent mechanism. PMID- 19186033 TI - QuadFit--a new cross-platform computer program for simulation of NMR line shapes from solids with distributions of interaction parameters. AB - A new Java computer program called QuadFit has been written to simulate NMR line shapes from solid materials. The program takes into account the major interactions, with a key feature that distributions of isotropic chemical shift and quadrupolar interaction parameters can be calculated, which are often encountered in amorphous and disordered materials. The quadrupolar interaction can be simulated for all the transitions for both half-integer and integer spins. The utility of the program is demonstrated with examples of (27)Al (nuclear spin I=5/2) in an atomically disordered aluminoborate mullite, (65)Cu (I=3/2) in CuInSe(2) and (10)B (I=3) in amorphous B(2)O(3). The program has good cross platform compatibility and is written for high stability. The program has been designed with an easy to use graphical interface. It can be run efficiently on any reasonably powerful PC and is freely available from the Warwick website (http://go.warwick.ac.uk/quadfit). PMID- 19186034 TI - In situ high temperature MAS NMR study of the mechanisms of catalysis. Ethane aromatization on Zn-modified zeolite BEA. AB - Ethane conversion into aromatic hydrocarbons over Zn-modified zeolite BEA has been analyzed by high-temperature MAS NMR spectroscopy. Information about intermediates (Zn-ethyl species) and reaction products (mainly toluene and methane), which were formed under the conditions of a batch reactor, was obtained by (13)C MAS NMR. Kinetics of the reaction, which was monitored by (1)H MAS NMR in situ at the temperature of 573K, provided information about the reaction mechanism. Simulation of the experimental kinetics within the frames of the possible kinetic schemes of the reaction demonstrates that a large amount of methane evolved under ethane aromatization arises from the stage of direct ethane hydrogenolysis. PMID- 19186035 TI - Dewetting pattern and stability of thin xyloglucan films adsorbed on silicon and mica. AB - Thin polysaccharide films prepared with xyloglucan (XG), a neutral polysaccharide extracted from the seeds of Guibourtia hymenifolia were prepared by spin-coating and drop deposition under pH3, pH5 and pH12, on silicon and mica substrates. Atomic force microscopy (AFM) images show flat nanoporous matrices with additional grain-like structures on both mica and silicon for pH 3 and pH 5. However, X-ray photoelectron spectroscopy (XPS) and Auger spectra of these adsorbed biopolymers prepared under alkaline condition (pH 12) reveal that Na(+) ions from the solution interact with the mica substrate surface and with XG forming chemical bonds. Both XPS and Auger results suggest XG depolymerisation during adsorption, caused by an alkaline ss-base catalyzed degradation mechanism, which is consistent with the more basic character of the mica surface under these conditions. Thus, the polysaccharide diffusion is inhibited during dewetting due to the surface bonding. On the other hand, the interaction of Na(+) in solution with the silicon surface is weaker, favoring its interaction with the polysaccharide, conserving the overall polymer structure of XG and allowing the biopolymer to slip and diffuse during dewetting, forming the final branched fractal structure. PMID- 19186036 TI - Dissimilar pH-dependent adsorption features of bovine serum albumin and alpha chymotrypsin on mica probed by AFM. AB - We studied bovine serum albumin (BSA) and alpha-chymotrypsin adsorption onto mica surfaces over a large pH range by atomic force microscopy (AFM) measurements in liquid. Data analyses (height, roughness and roughness factor) brought new insights on the conformation of proteins in soil environments, with mica as a model of soil phyllosilicates and non-hydrophobic surfaces. Validation of AFM approach was performed on BSA, whose behavior was previously described by nuclear magnetic resonance and infra-red spectroscopic methods. Maximum adsorption was observed near the isoelectric point (IEP). A stronger interaction and a lower amount of adsorbed proteins were observed below the IEP, which contrasted with the progressive decrease of adsorption above the IEP. We then studied the adsorption of alpha-chymotrypsin, a proteolytic enzyme commonly found in soils. AFM pictures demonstrated a complete coverage of the mica surface at the IEP in contrast to the BSA case. Comparison of the AFM data with other indirect methods broadened the understanding of alpha-chymotrypsin adsorption process through the direct display of the protein adsorption patterns as a function of pH. PMID- 19186037 TI - Pullulan acetate nanoparticles prepared by solvent diffusion method for epirubicin chemotherapy. AB - Pullulan acetate (PA) was synthesized by the reaction of pullulan with acetic anhydride in the presence of pyridine. PA was characterized by Fourier transform infrared (FT-IR) and proton nuclear magnetic resonance ((1)H NMR). A solvent diffusion method was employed in the current work to prepare PA nanoparticles. This technique had some advantages compared with other methods. The particle size increased from 185.7 nm to 423.0 nm with the degree of acetylation increasing from 2.71 to 3.0. Drug-loaded PA nanoparticles were prepared for controlled release of epirubicin (EPI). The drug entrapment and drug content increased with the degree substitution of PA increasing. EPI was released from the nanoparticles in a biphasic profile with a fast release rate in the first 10h followed by a slow release in vitro. A higher cytotoxicity against KB cells was found for EPI loaded PA nanoparticles in comparison with free EPI. Confocal laser scanning microscopy (CLSM) observations indicate that EPI-loaded nanoparticles were internalized and released in the cytoplasmic compartment. PMID- 19186038 TI - [Acute RSV bronchiolitis: should we be looking for pertussis?]. PMID- 19186039 TI - Various dietary polyunsaturated fatty acids modulate acrylamide-induced preneoplatic urothelial proliferation and apoptosis in mice. AB - To investigate whether various dietary fats affected preneoplatic lesions of urinary tract in acrylamide (ACR)-treated mice. Eighty Kunming mice were initiated with ACR at dose 10mg/kg bw, and fed with different polyunsaturated fatty acid (PUFA): 6% fish oil (enriched in n-3 PUFA), 6% corn oil (enriched in n 6), 6% olein (enriched in n-9, an EFA deficiency inducer), 10% fish oil, 10% corn oil, 10% olein and a commercial chow. Animals were autopsied 22 weeks. The liver fatty acid profile showed a close correlation with dietary sources, exhibiting macroscopic and biochemical EFA-deficient (EFAD) characteristics in ACR mice with olein. The frequency of simple urothelial hyperplasia (H) and dysplasia/carcinoma in situ (D/CIS) was significantly higher in ACR mice with corn oil or olein compared to ACR mice with commercial chow. Proliferation and abnormal luminal localized mitosis, also expression of proliferating cell nuclear antigen (PCNA), significantly increased in ACR mice with corn oil and olein than in ACR mice with commercial chow; moreover, abnormal apoptotic/mitosis ratio, expression of caspase-3, decreased in ACR mice with both olein and corn oil. Fish oil took no significant effect on almost all the parameters in ACR mice in this study. Results suggest that dietary PUFA modulate preneoplastic proliferation in ACR mice; n-6 PUFA (corn oil) and EFAD status (n-9 PUFA) exhibits a promoting activity; whereas, fish oil, rich in n-3 fatty acids, exhibits somewhat attenuated effect, and needs further research. PMID- 19186040 TI - Obesity and NK cells affect the expression of the long form of the leptin receptor Ob-Rb in liver of F344 rats. AB - In obesity, the regulatory effects of leptin, a primarily adipocyte-derived hormone, are severely disturbed affecting the control of energy homeostasis and immune functions. In addition, recent studies indicate that specific immune cells can affect glucose and lipid metabolism of liver. However, the contribution of body weight and immune cells, such as Natural Killer (NK) cells, to the regulation of the leptin-receptor expression remains elusive. Therefore, we investigated the expression of the signal-transducing long form of the leptin receptor (Ob-Rb) in diet-induced obesity and after adoptive cross-over NK cell transfer between normal weight and obese male F344 rats. Expression of Ob-Rb was significantly increased in liver in diet-induced obese rats as compared to normal weight littermates. Similarly, the expression of Ob-Rb was higher in liver of obese animals that received NK cells from either obese or normal weight donors as compared to normal weight animals that received NK cells from normal weight donors. Interestingly, normal weight animals that were transferred with NK cells from obese donors also showed a tendency towards a higher Ob-Rb expression. In contrast to the findings in liver, the expression of Ob-Rb in spleen or lung remained unaffected by changes in body weight or cross-over NK cell transfer. Our results suggest that the expression of Ob-Rb mRNA in liver, but not in spleen or lung, is dependent on the body weight but can also be influenced by NK cells, thereby indicating a bidirectional cross-talk between the metabolic and the immune system. PMID- 19186041 TI - Dose- and time-dependent effects of luteolin on carbon tetrachloride-induced hepatotoxicity in mice. AB - Carbon tetrachloride (CCl(4)) is a well-known model compound for producing chemical hepatic injury. This study investigated the protective effects of the flavonoid luteolin on the CCl(4)-induced hepatotoxicity in mice. Luteolin dissolved in dimethyl sulfoxide (DMSO) was administered intraperitoneally (i.p.) at 5 or 50 mg/kg as a single dose, and once daily for 2 consecutive days. Two hours after the final treatment, the mice were treated with CCl(4) (20 mg/kg, i.p.). CCl(4)-induced hepatotoxicity was reduced in a dose- and time-dependent manner, as determined by decreased serum aminotransferase activities and liver histopathology. CCl(4) intoxication resulted in an overexpression of heat shock protein gp96 in the mice liver, which was strongly attenuated by luteolin pretreatment. Luteolin has also decreased oxidative stress produced by CCl(4), as suggested by improvement in the Cu/Zn superoxide dismutase activity. The effect of luteolin on myeloperoxidase, an indicator of inflammatory cell infiltration, was also investigated. Treatment of the mice with luteolin resulted in a significant decrease in the myeloperoxidase activity. The hepatoprotective effect of luteolin against CCl(4) hepatotoxicity was higher in animals pretreated with luteolin for 2 consecutive days. This suggests that the protection might be due to induction of some adaptive mechanisms. The data indicate that luteolin could be effective in protecting mice from the hepatotoxicity produced by CCl(4). PMID- 19186042 TI - Adipose tissue, liver and pancreas structural alterations in C57BL/6 mice fed high-fat-high-sucrose diet supplemented with fish oil (n-3 fatty acid rich oil). AB - Fish oil treatment was used in reversing the morphological and metabolic changes of C57BL/6 mice fed high-fat-high-sucrose (HFHS) diet. Two-month-old male C57BL/6 mice were fed HFHS chow or standard chow (SC). At 3 months of age, HFHS mice were separated into an untreated group (HFHS) and a group treated with fish oil (HFHS Fo, 1.5g/kg/day). At 4 months of age, HFHS fed mice had an increase in body mass (BM) and total body fat, when the animals were sacrificed. Both parameters were lower in HFHS-Fo than in HFHS mice. Plasma glucose and insulin levels were not affected among the groups, but HFHS and HFHS-Fo animals had higher homeostasis model assessment for insulin resistance HOMA-IR ratio. HFHS and HFHS-FO mice had increased plasma total cholesterol and LDL-C, HFHS-Fo increased plasma HDL-C and decreased triglycerides levels. The liver mass (LM) and the adipocytes' size were larger in HFHS mice, while HFHS-Fo mice had a lower LM and smaller adipocytes. The liver steatosis and hepatocyte binucleation were increased in HFHS mice, while HFHS-Fo mice had reduced liver steatosis and hepatocyte binucleation. HFHS Fo mice had a lower pancreas mass, while HFHS animals had higher islet pancreatic diameter. The SC group showed strong expression for insulin, glucagon and a glucose transporter type 2 GLUT-2 in all pancreatic islets, while in HFHS mice there was less expression for GLUT-2. However, HFHS-Fo mice showed an increase of GLUT-2 expression. In conclusion, dietary fish oil treatment reduces body mass and fat pad adiposity, and also by reducing plasma TG and pancreatic islet hypertrophy in mice fed high-fat-high-sucrose diet. Furthermore, fish oil improves glucagon and GLUT-2 expressions when it is decreased in insulin, but in hepatocyte binucleation and hepatic steatosis where the effect is reduced. PMID- 19186043 TI - Luteal cell steroidogenesis in relation to delayed embryonic development in the Indian short-nosed fruit bat, Cynopterus sphinx. AB - The primary aim of this study was to determine the possible cause of slow or delayed embryonic development in Cynopterus sphinx by investigating morphological and steroidogenic changes in the corpus luteum (CL) and circulating hormone concentrations during two pregnancies of a year. This species showed delayed post implantational embryonic development during gastrulation of the first pregnancy. Morphological features of the CL showed normal luteinization during both pregnancies. The CL did not change significantly in luteal cell size during the delay period of the first pregnancy as compared with the second pregnancy. The circulating progesterone and 17beta-estradiol concentrations were significantly lower during the period of delayed embryonic development as compared with the same stage of embryonic development during the second pregnancy. We also showed a marked decline in the activity of 3beta-hydroxysteroid dehydrogenase, P450 side chain cleavage enzyme, and steroidogenic acute regulatory peptide in the CL during the delay period. This may cause low circulating progesterone and estradiol synthesis and consequently delay embryonic development. What causes the decrease in steroidogenic factors in the CL during the period of delayed development in C. sphinx is under investigation. PMID- 19186044 TI - The effect of c-di-GMP (3'-5'-cyclic diguanylic acid) on the biofilm formation and adherence of Streptococcus mutans. AB - Depending on a biofilm lifestyle, Streptococcus mutans (S. mutans) is thought to be one of the primary causative agents of dental caries. Biofilm formation and adhesion are crucial physiological functions and virulence factors for S. mutans. Thus, attempts to control the development of dental caries only by inhibiting one of the several virulence factors are not effective. Cyclic diguanylate (c-di-GMP) [bis(3',5')-cyclic diguanylic acid] is a prokaryotic cyclic dinucleotide second messenger that has been implicated in determining the timing and amplitude of complex biological processes from biofilm formation and virulence to photosynthesis. Here, we demonstrate that this signaling molecule also plays a role in the ability of S. mutans to initiate biofilm formation and adhere to tooth surfaces. To test this hypothesis, S. mutans UA159 and its gcp gene knockout mutant were assayed for their ability to initiate biofilm formation and adherence. The spatial distribution and architecture of the biofilms were examined by scanning electron microscopy. These results show that inactivation of the gcp gene resulted in the formation of an abnormal biofilm. We confirmed that c-di-GMP was effective in preventing biofilm formation of S. mutans UA159. We also found that extracellular c-di-GMP inhibited the adherence of S. mutans to tooth surfaces and reduced (>50%) biofilm formation compared to the untreated control. These results indicate that c-di-GMP attenuates the caries-inducing virulence factors of S. mutans. This suggests that c-di-GMP may be used alone or in combination with other antimicrobial agents, and that such a treatment could be developed into a novel method to prevent tooth decay. PMID- 19186045 TI - Role of protein kinase C in 15-HETE-induced hypoxic pulmonary vasoconstriction. AB - The aim of the present study was to investigate the roles of protein kinase C (PKC) signal transduction pathway in the 15-hydroxyeicosatetraenoic acid (15 HETE)-induced down-regulation expression of K(V) 1.5, K(V) 2.1 and K(V) 3.4, and pulmonary vasoconstriction under hypoxia. Tension measurements on rat pulmonary artery (PA) rings, Western blots, semi-quantitative PCR and whole-cell patch clamp technique were employed to investigate the effects of 15-HETE on PKC and K(V) channels. Hypericin (6.8 micromol/L), a PKC inhibitor, significantly attenuated the constriction of PA rings to 15-HETE under hypoxia. The down regulation of K(V) 1.5, K(V) 2.1 and K(V) 3.4 channel expression and inhibition of whole-cell K currents (I(K)(V)) induced by 15-HETE were rescued and restored, respectively, by hypericin. These studies indicate that the PKC signal transduction pathway is involved in 15-HETE-induced rat pulmonary vasoconstriction under hypoxia. 15-HETE suppresses the expression of K(V) 1.5, K(V) 2.1 and K(V) 3.4 channels and inhibits I(K)(V) through the PKC signaling pathway in pulmonary arterial smooth muscle cells. PMID- 19186046 TI - Elevated plasma TGF-beta1 levels in patients with chronic obstructive pulmonary disease. AB - BACKGROUND: Transforming growth factor-beta(1) (TGF-beta(1)), a multifunctional cytokine, has been implicated to be responsible for the increased deposition of extracellular matrix in the airways, and increased submucosal collagen expression in chronic obstructive pulmonary disease (COPD). We determined plasma TGF-beta(1) levels in patients with COPD and explored its association with common functional polymorphisms of TGF-beta(1) gene at C-509T and T869C in the development of COPD in a case-control study. METHODS: Stable COPD patients who were ever smokers, and age and pack-years smoked matched healthy controls (n = 205 in each group) were recruited for measurement of plasma TGF-beta(1) levels using commercially available ELISA kit, and genotyped at C-509T and T869C functional polymorphisms of TGF-beta(1) gene using polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP). RESULTS: COPD patients had significantly elevated plasma TGF-beta(1) levels in comparison to healthy controls irrespective of the genotypes. Allele frequencies and genotype distributions at both polymorphic sites were not different among COPD patients or controls. TGF-beta(1) levels were inversely correlated (Pearson's correlation analysis) with FEV(1) (% predicted) (p < 0.001) and FVC (% predicted) (p < 0.001). CONCLUSION: The findings of elevated plasma TGF-beta(1) levels in patients with COPD suggest that TGF-beta(1) may play a role in COPD pathogenesis. The C-509T and T869C functional polymorphisms of TGF-beta(1) gene do not represent a genetic predisposition to COPD susceptibility in Hong Kong Chinese patients. PMID- 19186048 TI - A multiple-ligand approach to extending the dynamic range of analyte quantification in protein microarrays. AB - This work describes a concept for extending the dynamic range of quantification in an affinity biosensor assay by using a set of ligands with different affinities toward a common analyte. Three synthetic, biotinylated polypeptides capable of binding a model protein analyte with different affinities (10(-9) M < or = K(d) < or = 10(-7) M) were immobilized in a microarray format on a gold slide covered with an oligo(ethylene glycol)-containing alkane thiolate self assembled monolayer. A five-element affinity array, comprising single-peptide spots as well as spots where peptides were immobilized in mixtures, was realized by means of piezodispensation. Imaging surface plasmon resonance was used to study binding of the analyte to the different spots. The lower limit of analyte quantification was approximately 3 nM and the corresponding upper limit was increased by more than an order of magnitude compared to if only the highest affinity ligand would have been used. Affinity array sensors with multiple ligands for each analyte are particularly interesting for omitting dilution steps and providing highly accurate data in assays where several analytes such as disease biomarkers with extremely variable concentrations are quantified in parallel. PMID- 19186047 TI - Actin and endocytosis: mechanisms and phylogeny. AB - The regulated assembly of actin filament networks is a crucial part of endocytosis, with crucial temporal and spatial relationships between proteins of the endocytic and actin assembly machinery. Of particular importance has been a wealth of studies in budding and fission yeast. Cell biology approaches, combined with molecular genetics, have begun to uncover the complexity of the regulation of actin dynamics during the endocytic process. In a wide range of organisms, clathrin-mediated endocytosis appears to be linked to Arp2/3-mediated actin assembly. The conservation of the components, across a wide range eukaryotic species, suggests that the partnership between endocytosis and actin may be evolutionarily ancient. PMID- 19186049 TI - Pediatric malignancies: update on sarcomas and leukemia development in children. AB - Despite the rarity of malignancies in children, their study has provided important insights into normal cellular growth regulation and into cancer development. Here we present a few vignettes in pediatric leukemia and sarcoma that highlight a number of trends in basic, translational, and clinical research. These include the application of new methodologies for oncogene discovery, new approaches to study oncogenes, the use of genetically engineered models to identify tumor cell-of-origins, and the use of targeted therapeutics against pathways not thought to be mutant in tumor cells. Through the use of such methodologies, an improved understanding of pediatric malignancy is emerging, and is being applied to the care of children with cancer. PMID- 19186050 TI - Understanding and engineering RNA sequence specificity of PUF proteins. AB - PUF proteins are RNA-binding proteins named for founding members PUMILIO and fem 3 binding factor (FBF). Together these proteins represent the range of known RNA recognition properties. PUMILIO is a prototypical PUF protein whose RNA sequence specificity is simple, elegant, and predictable. FBF displays differences in RNA recognition that represent divergence from the prototype. Here we review recent studies that examine the engineering of sequence specificity of PUF proteins and its applications as well as studies that increase our understanding of the natural diversity of RNA recognition by this family of proteins. PMID- 19186051 TI - Phase II study of weekly Kahalalide F in patients with advanced malignant melanoma. AB - This phase II clinical trial evaluated the antitumour response of Kahalalide F (KF) 650 microg/m(2) given as a 1-h weekly infusion in advanced malignant melanoma patients, both untreated and those who relapsed or progressed after one line of systemic therapy. Of 24 enrolled patients (median age, 55 years; range, 28-89), 14 patients had been previously treated with chemotherapy or biological therapy. No RECIST responses occurred; five chemotherapy-naive patients with cutaneous melanoma had disease stabilisation for > or = 3 months; median progression-free survival was 1.7 months (95% CI, 1.2-1.9 months); and median overall survival was 10.8 months (95% CI, 5.0-upper limit not reached). The most common laboratory toxicities were non-cumulative increase of transaminases (ALT/AST) and gamma-glutamyltransferase (GGT). No patients experienced leukopenia and thrombocytopenia during the study. KF was a well-tolerated and safe chemotherapy regimen. Despite a favourable safety profile, this trial was closed after the first stage because of the lack of objective response in patients with malignant melanoma. PMID- 19186052 TI - Biosorption of phenol and 2-chlorophenol by Funalia trogii pellets. AB - The removal of phenol (Ph) and 2-chlorophenol (2-CPh) from aqueous solution by native and heat inactivated fungus Funalia trogii pellets were investigated. The effects of contact time, solid/liquid ratio, optimum pH and temperature on the phenols removal capacity by the pellets were established. The removal efficiency of phenols increased significantly with increasing biomass dose. The optimum pH was detected to be 8.0. The second-order equations are described and evaluated on the basis of a comparative estimation of the corresponding coefficients. The phenol removal equilibrium isotherm was modeled by the Langmuir equations. The enthalpy change values were obtained between -7.62 and -10.64 kJ/mol. This indicated that the uptake of phenols either on native or heat inactivated fungal pellets was based on a physical adsorption process. PMID- 19186053 TI - Simultaneous removal of organic matter and nitrogen compounds by combining a membrane bioreactor and a membrane biofilm reactor. AB - Hydrogen-based membrane biofilm reactors (MBfR) have been applied to the denitrification of nitrate-containing water and wastewater. Adding an aerobic membrane bioreactor (MBR) to a MBfR provides significant nitrification and organic oxidation because most wastewater also contains a significant concentration of organic material and ammonium nitrogen. This study describes experiments that investigate the removal of organic and nitrogenous compounds in the combined MBR/MBfR system. The experiments demonstrate that the MBR/MBfR combination successfully performs COD oxidation and nitrogen removal for organic and ammonium loads in the ranges of 1000-4300gCOD/m(3)-d and 200-230gN/m(3)-d, respectively. Total-nitrogen removal was controlled by nitrification in the MBR, because the MBfR denitrified all of the NO(3)(-) provided by the MBR. The nitrate flux in the MBfR was in the range of 4-8gN/m(2)-d for cases of almost complete denitrification (>99 %); the H(2) flux was varied from 1.4 to 2.8gH(2)/m(2)-d. PMID- 19186054 TI - Drin pesticides removal from aqueous solutions using acid-treated date stones. AB - This work describes the potential applicability of chemically and thermally treated date stones for removing drin pesticides (aldrin, dieldrin and endrin) from aqueous solutions. The effect of several parameters, such as sorbent particle size, adsorbent dose, shaking speed, shaking time, concentration of pesticide solution and temperature, was evaluated by batch experiments. Pesticide determination was carried out using stir bar sorptive extraction and gas chromatography coupled with mass spectroscopy. Maximum removal efficiency (93%) was reached using 0.1 g of acid-treated date stones (ATDS) (63-100 microm) and 100 mL of aldrin (0.5 mg L(-1)). The removal efficiency of drin pesticides decreased in the order of aldrin, dieldrin and endrin, and decreased as the temperature rose. Adsorption data were processed according to various kinetic models. Lagergren and Morris-Weber equations were applied to fit the kinetic results. The second order model was the most suitable on the whole, and intra particle diffusion was found to be the rate-controlling the adsorption process. According to adsorption kinetic data, 3.5h were considered as the equilibrium time for determining adsorption isotherms. Adsorption data were analyzed by the Langmuir, Freundlich and Dubinin-Radushkevich adsorption approaches. Experimental results showed that the Freundlich isotherm model best described the adsorption process. In addition, thermodynamic parameters such as DeltaH, DeltaS and DeltaG were calculated. Negative values of DeltaH and DeltaG indicate the exothermic and spontaneous nature of pesticide adsorption on ATDS. PMID- 19186055 TI - The identification of orally bioavailable thrombopoietin agonists. AB - Recently, we disclosed a series of potent pyrimidine benzamide-based thrombopoietin receptor agonists. Unfortunately, the structural features required for the desired activity conferred physicochemical properties that were not favorable for the development of an oral agent. The physical properties of the series were improved by replacing the aminopyrimidinyl group with a piperidine-4 carboxylic acid moiety. The resulting compounds possessed favorable in vivo pharmacokinetic properties, including good bioavailability. PMID- 19186056 TI - A thiabendazole sulfonamide shows potent inhibitory activity against mammalian and nematode alpha-carbonic anhydrases. AB - A sulfonamide derivative of the antihelmintic drug thiabendazole was prepared and investigated for inhibition of the zinc enzyme carbonic anhydrase CA (EC 4.2.1.1). Mammalian isoforms CA I-XIV and the nematode enzyme of Caenorhabditis elegans CAH-4b were included in this study. Thiabendazole-5-sulfonamide was a very effective inhibitor of CAH-4b and CA IX (K(I)s of 6.4-9.5nm) and also inhibited effectively isozymes CA I, II, IV-VII, and XII, with K(I)s in the range of 17.8-73.2nM. The high resolution X-ray crystal structure of its adduct with isozyme II evidenced the structural elements responsible for this potent inhibitory activity. PMID- 19186057 TI - Water-soluble prodrugs of an Aurora kinase inhibitor. AB - Compound 1 (SNS-314) is a potent and selective Aurora kinase inhibitor that is currently in clinical trials in patients with advanced solid tumors. This communication describes the synthesis of prodrug derivatives of 1 with improved aqueous solubility profiles. In particular, phosphonooxymethyl-derived prodrug 2g has significantly enhanced solubility and is converted to the biologically active parent (1) following iv as well as po administration to rodents. PMID- 19186058 TI - New and selective ryanodine receptor activators for insect control. AB - Diamide insecticides have emerged as one of the most promising new classes of insecticide chemistry owing to their excellent insecticidal efficacy and high margins of mammalian safety. Chlorantraniliprole and flubendiamide, the first two insecticides from this class, demonstrate exceptional activity across a broad range of pests in the order Lepidoptera. This chemistry has been confirmed to control insects via activation of ryanodine receptors which leads to uncontrolled calcium release in muscle. The high levels of mammalian safety are attributed to a strong selectivity for insect over mammalian receptors. PMID- 19186059 TI - Synthesis and antiprotozoal activity of novel 1-methylbenzimidazole derivatives. AB - In this paper are reported the synthesis and antiprotozoal activity in vitro of 24 1-methylbenzimidazole derivatives (13-36) substituted at position 2 with aminocarbonyl, N-methylaminocarbonyl, N,N-dimethylaminocarbonyl, ethoxycarbonyl, 1-hydroxyethyl and acetyl groups, some of them with chlorine atoms at the benzenoid ring. Compounds 13-36 were more active than metronidazole, the choice drug against Giardia intestinalis and most of them against Trichomonas vaginalis. The most active group of compounds for both parasites was that with a 2 ethoxycarbonyl group (16, 22, 28, 34), independently of the substitution pattern at the benzenoid ring. PMID- 19186060 TI - Insect neuropeptides: structures, chemical modifications and potential for insect control. AB - Insect neuropeptides are involved in almost all physiological processes in insects, such as diuresis, ecdysis, pheromone biosynthesis and control of muscle activity. Thus, these small peptide hormones and their receptors are promising targets for a novel generation of selective and non-neurotoxic insecticides. However, due to poor bioavailability, pharmacokinetics and short half-life the peptides themselves cannot be used as insect control agents. The past two decades have seen an increase in research into the discovery of non-peptide small molecules that function as mimics for neuropeptides. This review presents an overview on structure-activity studies, conformational analyses and peptidomimetic modifications of selected insect neuropeptides with a special potential for application in pest control. PMID- 19186061 TI - Synthesis and biological testing of thioalkane- and thioarene-spaced bis-beta-D glucopyranosides. AB - A three-step synthesis of bis-beta-D-glucopyranosides containing thioalkane or thioarene spacers of different length and flexibility is described. The key-step reaction allows an easy modulation of final saccharidic products so that a library of molecules with different glycosidic residues and spacers can be obtained. Two of the new thioarene-spaced bis-beta-D-glucopyranosides endow with a specific cytotoxic potential. A more detailed investigation of one of the two compounds ascertains that this effect is attributable to induction of cell death by apoptosis. PMID- 19186062 TI - Synthesis and biological evaluation of fused thio- and selenopyrans as new indolocarbazole analogues with aryl hydrocarbon receptor affinity. AB - A series of thio- and selenopyrans having two fused indole units, structurally related to indolocarbazoles, have been prepared and evaluated for aryl hydrocarbon receptor (AhR) affinity, leading to the identification of several new significant AhR ligands. In particular, the parent thiopyrano[2,3-b:6,5 b']diindole and its derivative having a methyl group in the central ring, as well as the two corresponding selenopyrans, displayed the highest potencies of the compounds tested. PMID- 19186063 TI - An improved radiosynthesis of the muscarinic M2 radiopharmaceutical, [18F]FP TZTP. AB - The radioligand 3-(4-(3-[(18)F]fluoropropylthio)-1,2,5-thiadiazol-3-yl)-1-methyl 1,2,5,6-tetrahydropyridine ([(18)F]FP-TZTP) is an agonist with specificity towards subtype 2 of muscarinic acetylcholine (M2) receptors. It is currently the only radiotracer available for imaging M2 receptors in human subjects with positron emission tomography. The present study reports on an improved method for the synthesis of [(18)F]FP-TZTP, automated using a GE TRACERlab FX(FN) radiosynthesis module. A key facet was the use of a new precursor, 3-(4-(1-methyl 1,2,5,6-tetrahydropyridin-3-yl)-1,2,5-thiadiazol-3-ylthio)propyl 4 methylbenzenesulfonate. The precursor was fluorinated via nucleophilic displacement of the tosyloxy group by potassium cryptand [(18)F]fluoride (K[(18)F]/K(222)) in CH(3)CN at 80 degrees C for 5 min, and purified by HPLC. Formulated [(18)F]FP-TZTP was prepared in an uncorrected radiochemical yield of 29+/-4%, with a specific activity of 138+/-41 GBq/micromol (3732+/-1109 mCi/micromol) at the end of synthesis (35 min; n=3). This methodology offers higher yields, faster synthesis times, an optimized precursor, and simpler automation than previously reported. PMID- 19186064 TI - Estimation of heat generation by radioactive decay of some phosphate rocks in Egypt. AB - Radiogenic heat production data for phosphate rocks outcropping on the three main areas Eastern Desert, Western Desert and Nile Valley are presented. They were derived from uranium, thorium and potassium concentration measurements of gamma radiation originating from the decay of (214)Bi ((238)U series), (208)Tl ((232)Th series) and the primary decay of (40)K. A low radioactive heat production rate (0.32+/-0.1 microWm(-3)) was found for Wadi Hegaza, whereas the highest value (19+/-4.1 microWm(-3)) was found for Gabel Anz, Eastern Desert of Egypt. PMID- 19186065 TI - Spallation neutron emission spectra in medium and heavy target nuclei by a proton beam up to 140 MeV energy. AB - In this study, neutron-emission spectra produced by (p,xn) reactions for some spallation neutron target nuclei such as (65)Cu, (204,206,207,208)Pb, (209)Bi, (232)Th and (238)U have been calculated by a proton beam up to 140 MeV. Multiple pre-equilibrium mean free path constants from internal transition for (204,208)Pb (p, xn) neutron emission spectra have been investigated. In the calculations, pre equilibrium effects were calculated by using new evaluated hybrid model and geometry dependent hybrid model, full exciton model and cascade exciton model. The reaction equilibrium component was calculated by Weisskopf-Ewing model. The obtained results have been discussed and compared with the available experimental data and found agreement with each other. PMID- 19186066 TI - Separation of the 90Sr-90Y pair with cerium(IV) iodotungstate cation exchanger. AB - A new inorganic ion exchanger, cerium(IV) iodotungstate, has been synthesized and characterized by elemental analysis and IR spectroscopy. This inorganic material is very stable thermally, chemically and with respect to radiation. Its ion exchange capacity for alkali and alkaline earth metal ions has been studied. (90)Y has been successfully separated from its parent (90)Sr in the equilibrium mixture with this exchanger at pH 6.0 with 0.0063 M EDTA solution as an eluent. PMID- 19186068 TI - Clinical application of 3D pre-bent titanium implants for orbital floor fractures. AB - INTRODUCTION: Orbital structures are affected in approximately 40% of all cases of craniofacial trauma. Changes in the bony orbital dimensions can alter the function of intraorbital contents and lead to serious complications. The unique anatomy of the orbit and the resulting surgical approaches make the process of fitting and aligning implants difficult, time consuming and operator dependent. It is now possible to make relatively inexpensive anatomical models on the basis of computed tomography images, using rapid prototyping. Such models can be used as templates to form titanium mesh implants, which are then used in the reconstruction of orbital floor defects. MATERIAL AND METHODS: Six patients with facial trauma were included in this study. First, 3D virtual models and then physical models were created. These were used as templates to shape the titanium mesh and then intraoperatively as guides to aid correct implant placement in the orbit. RESULTS: Significant improvement resulted in three cases and total recovery in three cases. CONCLUSION: It is financially viable to build anatomical models, on the basis of CT studies, that can be used in the repair of orbital floor fractures. PMID- 19186067 TI - Targeting prokaryotic choline oxidase into chloroplasts enhance the potential of photosynthetic machinery of plants to withstand oxidative damage. AB - Chloroplasts from plants of transgenic lines expressing prokaryotic choline oxidase gene (the codA(ps) gene; GenBank accession number-AY589052) and wild-type of chickpea and Indian mustard were evaluated for their efficacy to withstand photoinhibitory damage, by exposing them to high light intensity ( approximately 1200micromolm(-2)s(-1) photon flux density) at 10 and 25 degrees C. Western analysis confirmed presence of choline oxidase in chloroplasts of only transgenic lines. The loss in PS II activity in chloroplasts of wild-type exposed to high light intensity was significantly higher than that in chloroplasts of transgenic chickpea as well as Indian mustard. Although, chloroplasts of both wild-type and transgenic chickpea as well as Indian mustard were more sensitive to photoinhibitory damage at 10 than at 25 degrees C, the damage recorded in chloroplasts harboring choline oxidase was significantly lower than those of wild type. High light promotes H(2)O(2) production in chloroplasts more significantly at low temperature (10 degrees C) than at 25 degrees C. We compared low temperature accelerated photoinhibition of chloroplasts with that caused due to exogenously applied H(2)O(2). Although exogenous H(2)O(2) accelerated high light intensity induced loss in PS II activity of chloroplasts of wild-type, it caused only a little alteration in PS II activity of chloroplasts from transgenic lines of both chickpea and Indian mustard, demonstrating that the chloroplasts harboring choline oxidase are better equipped to resist photoinhibition. We hypothesize that H(2)O(2) produced by choline oxidase as a byproduct during synthesis of glycinebetaine is responsible for building stronger antioxidant system in chloroplasts of transgenic lines compared to that of wild-type. PMID- 19186069 TI - Segmentectomy for carcinoid arising from an accessory cardiac bronchus. PMID- 19186071 TI - Priming of cardiopulmonary bypass: effect on renal function. PMID- 19186072 TI - Hypoxia-inducible factor in cancer angiogenesis: structure, regulation and clinical perspectives. AB - Tumor hypoxia is a common feature of many cancers. A master regulator of hypoxic response is the transcription factor hypoxia-inducible factor-1 (HIF-1). It functions as a master regulator of oxygen and undergoes conformational changes in response to varying oxygen concentrations. In this paper, we review what has been described about HIF-1: its structure, its regulation and target genes, its role in cancer, and its implication for cancer therapy. PMID- 19186073 TI - 1Alpha,25-dihydroxyvitamin D3 inhibits CD40L-induced pro-inflammatory and immunomodulatory activity in human monocytes. AB - CD40 ligand (CD40L) stimulation induces proinflammatory and immunomodulatory activity in monocytes. Here, we report on the effects of the steroid hormone 1alpha,25-dihydroxyvitamin D3 (1,25D3) on human blood monocytes that have been stimulated with the CD40L ligand. Co-treatment of CD40L-stimulated monocytes with 1,25D3 resulted in reduced production and secretion of tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta, as well as in reduced expression of the surface co-stimulatory molecules CD80 and CD86. In addition, costimulation of CD4+ T lymphocytes by monocytes co-treated with CD40L and 1,25D3 resulted in reduced cell proliferation and diminished interferon (IFN)-gamma but enhanced IL 10 production by CD4+ T cells. Finally, 1,25D3 interfered with the ability of CD40L to rescue monocytes from apoptosis induced by serum withdrawal. These findings suggest that 1,25D3 may regulate the interaction of monocytes with T cells or other cell types that express CD40L, thus influencing the outcome of the immune or inflammatory response. PMID- 19186074 TI - Detection of VDR gene ApaI and TaqI polymorphisms in patients with type 2 diabetes mellitus using PCR-RFLP method in a Turkish population. AB - Type 2 diabetes mellitus (T2DM) is by far the most common type of diabetes and is characterized by insulin resistance and altered insulin secretion. Some genes, such as the vitamin D receptor gene (VDR, NM_001017535; GI: 7421), involved in its metabolic pathway have been regarded as good candidates for T2DM. In this study, we investigated whether there was an association of VDR: g.59979G>T or c.1025-49G>T (ApaIG>T) and g.60058T>C or c.1056T>C (TaqIT>C) polymorphisms in the 3' untranslated region of VDR with T2DM in a Turkish population. We collected blood samples from 241 individuals (72 patients with T2DM and 169 healthy individuals), and their DNA was isolated. Polymorphisms of the VDR were analyzed by DNA amplification with polymerase chain reaction and endonuclease digestion with ApaI and TaqI. Body mass index was higher in T2DM patients than in control individuals. However, the frequency of g.59979TT genotype in T2DM patients was not significantly increased compared to healthy subjects (37.5% vs. 36.1%, respectively). Although the VDR g.60058CC genotype in T2DM patients (19.4%) was higher than that in healthy individuals (11.2%), there was no significant difference. In the same way, there was no difference between the groups in allele frequencies. In conclusion, our study did not provide evidence for the association of two examined VDR polymorphisms with T2DM in a Turkish population. PMID- 19186075 TI - Osteoid osteoma of the coronoid process with involvement of the proximal radioulnar joint: a case report. PMID- 19186076 TI - The anatomy of the deltoid insertion. AB - HYPOTHESIS: The deltoid muscle is in continuity with the distal arm fascia and musculature. MATERIALS AND METHODS: Ten fresh-frozen upper extremity cadaveric specimens were dissected to determine the insertional anatomy of the deltoid muscle. Measurements were made with micro-calipers and acetate grid sheets. The deltoid tendon and fibrous aponeurosis was in continuity with the lateral intermuscular septum posteriorly and the lateral aspect of the brachialis and deep brachial fascia anteriorly in all ten specimens. This interconnection remained in continuity following complete release of the deltoid insertion. RESULTS: The width of the insertion of the anterior head of the deltoid was a mean of 7.3 mm, the middle averaged 4.7 mm, and the posterior averaged 7.8 mm. The mean length of the anterior insertion was 70 mm, the middle was 48.4 mm, and the posterior was 63.4 mm. The mean width of the deltoid tendon and the investing fascia at the superior margin of the insertion was 21.9 mm and at the inferior margin 13.1 mm. DISCUSSION: Deltoid muscle integrity is critical to shoulder function. The deltoid insertion, however, is often partially released during surgical approaches for internal fixation of proximal humerus fractures and shoulder arthroplasty. Partial detachment without repair as performed during surgical approaches should not result in complete loss of continuity of the deltoid insertion. CONCLUSION: Knowledge of the distal insertion and interconnections of the deltoid allows for more anatomic repair following extended releases during complex fracture fixation or revision surgery. PMID- 19186077 TI - Functional outcome following endoprosthetic reconstruction of the proximal humerus. AB - BACKGROUND: The proximal humerus is a common site for tumors, either metastatic or primary. Thus it is a frequent site of intervention in musculoskeletal oncology surgery. We looked at the use of endoprosthetic reconstructions in surgical intervention for tumors of the proximal humerus. METHODS: A review of our database from 1990 to 2005 revealed 83 proximal humeral endoprosthetic reconstructions following an intra-articular, deltoid muscle, and axillary nerve sparing resection. Medical records and radiographs were reviewed to determine shoulder range of motion, MSTS scores, and any complications. The median patient age was 55 years (range, 13-80). The mean follow-up was 30 months (range, 1-199). RESULTS: Mean active abduction was 41 degrees (range, 10-90 degrees) and mean active forward flexion was 42 degrees (range, 5-115 degrees). The mean MSTS score was 63% (range, 40-83%). Implant-related complications included 2 deep infections and 22 patients with proximal migration of the prosthesis. No prostheses loosened. Only 2 required removal (1 for infection and 1 for progression of metastatic disease). CONCLUSIONS: A proximal humeral endoprosthesis provides a durable reconstruction with a relatively low complication rate. Although it provides a stable platform for elbow and hand function, actual shoulder function is limited. PMID- 19186078 TI - Partial humeral head resurfacing for osteonecrosis. AB - BACKGROUND: The purpose of this study is to present our experience treating patients in the advanced stages of osteonecrosis of the humeral head with partial resurfacing of the humeral head. METHODS: This is a prospective series of 12 shoulders in 11 patients diagnosed with osteonecrosis of the humeral head who underwent partial humeral head resurfacing. Their mean age was 56 years. Preoperative and postoperative standardized evaluations included history, physical examination, radiographs, and clinical scoring systems, including the Western Ontario Osteoarthritis of the Shoulder index, Shoulder Score Index derived from the American Shoulder and Elbow Surgeons evaluation form, Constant score, and score on the visual analog scale for pain. The mean follow-up was 30 months. RESULTS: Postoperatively, all patients reported significant pain relief. Scores on the visual analog scale for pain improved from 75 preoperatively to 16 postoperatively (P < .001). Physical examination showed significant improvements in functional outcomes as well. Forward elevation improved from a mean of 94 degrees preoperatively to 142 degrees postoperatively (P < .001). Good to excellent results were also observed for the Western Ontario Osteoarthritis of the Shoulder index, Shoulder Score Index, and Constant score. CONCLUSIONS: This prospective series on partial resurfacing of the humeral head for patients with advanced-stage osteonecrosis has shown it to be effective in relieving pain and restoring function. PMID- 19186079 TI - Missed posterior dislocation of the shoulder after intramedullary fixation of humeral fractures: a report of three cases. PMID- 19186080 TI - Severe chondrolysis after shoulder arthroscopy: a case series. AB - HYPOTHESIS: Chondrolysis has been observed after shoulder arthroscopy and results in severe glenohumeral complications. MATERIALS AND METHODS: Twenty three cases of post-arthroscopic glenohumeral chondrolysis, occurring between 2005-2006, are reported following a variety of arthroscopic shoulder procedures. Presenting complaints, signs and symptoms, associated operative findings, and potential etiological factors are reviewed. Management options are summarized. RESULTS: Of the 23 cases of chondrolysis identified in our practice over a two year period, 14 occurred in patients following labral repair using a bioabsorbable device. Seventeen of the 23 patients used a high volume intra-articular pain pump for 48 hours after surgery. Seven of the 23 cases had documented use of a thermal probe. Four cases occurred in shoulders with no reported use of fixation anchors, pain pumps, or thermal probes. All cases had at least a 20 cc intra-articular bolus injection of 0.25% bupivicaine with epinephrine. DISCUSSION: This case series identifies several common factors that could be responsible for post-arthroscopic glenohumeral chondrolysis. No single mechanism can be implicated based on the results of this study. Although strong concerns are raised over the use of intra articular local anesthetics, glenohumeral chondrolysis appears to be an unfortunate convergence of multiple factors that may initiate rapid dissolution of articular cartilage and degenerative changes. CONCLUSION: Chondrolysis is a devastating complication of arthroscopic shoulder surgery that can result in long term disabling consequences. Further research is required to specifically identify causative factors. Until this is a available, we strongly advise against the use of large doses of intra-articular placement of local anesthetics. PMID- 19186081 TI - Neuropsychological predictors of quality of life in focal epilepsy. AB - Spontaneous complaints of outpatients with focal epilepsy often stress the relationship between cognitive deficits and Quality of Life (QOL). Consequently, the aim of the present study was to find the best neuropsychological predictors of QOL in individuals with focal epilepsy, in order to guide their ambulatory health care. A sample of 71 Portuguese patients was studied: 40 female, 47 married, with a mean age of 37.48 years (S.D.=11.79, 16-62), mean education of 7.93 (S.D.=4.05, 3-17), and focal epilepsy of moderate severity. A Socio demographic and Clinical Questionnaire, the SF-36 v1, the Cognitive Functioning Scale from the ESI-55, a Seizure Control scale (items from the Liverpool Seizure Severity Scale), and several neuropsychological tests were used. Semantic Fluency was the only predictor of Physical Functioning, Role Functioning - Physical, and Mental Health; I.A. Test predicted Bodily Pain; and Attentive Matrices predicted General Health, Vitality, and Role Functioning - Emotional. The Mental Component of the SF-36 v1 was predicted by Attentive Matrices, and the Physical Component was predicted by Semantic Fluency. Cognitive Functioning was predicted by the Token Test. Social Functioning and Seizure Control presented no statistically significant correlation with the neuropsychological indicators used. These results underscore the importance of cognitive performance to the QOL of individuals with focal epilepsy, supporting the systematic screening of cognitive performance in this population. Additionally, they suggest cognitive rehabilitation has the potential to improve these individuals' QOL. PMID- 19186082 TI - Does RNA editing play a role in the development of urinary bladder cancer? AB - PURPOSE: A-to-I RNA editing is essential for the development of normal cells and is involved in a wide variety of biological pathways. Currently, limited information suggests linkage between changes in RNA editing levels and the development of cancer. We aimed to explore the possible linkage between altered RNA editing levels and the development of human urinary bladder neoplasms. MATERIALS AND METHODS: Thirty-two patients underwent transurethral resection of bladder tumor. Normal and tumoral urinary bladder tissues were obtained from each patient during surgery. Total RNA was extracted from tissue cells and converted by RT-PCR reaction to cDNA molecules for further analysis. We explored known editing sites in RNA encoding for proteins (BLCAP, Cyfip2, FLNA, GluB Q/R) as well as in RNA transcribed from Alu elements in noncoding regions of the genes encoding for CARD11, FANCC, MDM4, BRCA1, and RBBP9 proteins. Editing levels were determined using Sequenom MassARRAY Compact Analyzer. RESULTS: Eleven tumoral tissues obtained were low grade TCC, 14 high grade TCC, 1 CIS, and another 5 inflammation. One sample contained only normal tissue. We got a total number of 30 normal bladder tissue samples and overall 29 paired samples (i.e., normal and tumoral tissues obtained from the same patient). Statistical analysis revealed no significant changes in editing levels between normal and tumoral tissues. CONCLUSIONS: Relying on the results obtained for 9 different editing sites, it can be determined that RNA editing is an epigenetic mechanism that does not participate in the evolution of urinary bladder cancer. PMID- 19186083 TI - Diagnosis and management of restless legs syndrome in children. AB - Recent published evidence suggests that restless legs syndrome (RLS) and periodic limb movement disorder (PLMD) are common condition in children and adolescents. It is likely that if left untreated, RLS and PLMD may lead to adverse cardiovascular and neurocognitive consequences. However, the diagnosis of RLS and PLMD in children is challenging, particularly because children are relatively unable to describe typical RLS symptoms. The International Restless Legs Study Group has recently published consensus criteria for the diagnosis of RLS and PLMD in children. In addition to clinical description of RLS symptoms, supportive evidence including the presence of clinical sleep disturbances, documented periodic limb movements in sleep from overnight sleep study and family history of restless legs syndrome may be required. Few if any controlled studies have addressed the management of RLS and PLMD, which may involve both non pharmacologic and pharmacologic approaches. In this context, the importance of avoidance of aggravating factors and good sleep hygiene cannot be overemphasized. Children with evidence of low-iron storage, i.e., low-serum ferritin and/or iron levels may likely benefit from iron therapy. While there is overall limited experience regarding the use of dopaminergic agents in children with RLS and PLMD, published reports suggesting efficacy of compounds such as levodopa, ropinirole, pramipexole and pergolide have emerged. Other medications including benzodiazepine, anti-convulsants, alpha-adrenergic and opioid medications have not been adequately studied in children. Children with RLS and PLMD should have regular follow-up visits to evaluate clinical improvement and to monitor adverse effects from the selected therapy. Based on aforementioned findings, it is clear that a substantial research effort is needed to evaluate the pathophysiology, clinical presentation, treatment modalities, and overall long-term outcome of children with RLS and PLMD. PMID- 19186084 TI - The effects of isoflurane minimum alveolar concentration on withdrawal reflex activity evoked by repeated transcutaneous electrical stimulation in ponies. AB - The aim of this study was to quantify the effects of isoflurane at approximately the minimum alveolar concentration (peri-MAC) on the temporal summation (TS) of reflex activity in ponies. TS was evoked by repeated electrical stimulations applied at 5 Hz for 2 s on the digital nerve of the left forelimb of seven ponies. Surface electromyographic activity was recorded from the deltoid and common digital extensor muscles. TS thresholds and amplitude of response to stimulations of increasing intensities were assessed during anaesthesia at 0.85, 0.95 and 1.05 times the individual MAC, and after anaesthesia in standing animals. Under isoflurane anaesthesia, TS thresholds increased significantly in a concentration-dependent fashion and at each isoflurane MAC, the responses increased significantly for increasing stimulation intensities. A concentration dependent depression of evoked reflexes with a reduction in the slopes of the stimulus-response function was observed for both muscles. The results demonstrated that with this model it is possible to describe and quantify the effects of anaesthetics on spinal sensory-motor processing in ponies. PMID- 19186085 TI - Levetiracetam as monotherapy for seizures in a neonate with acute lymphoblastic leukemia. AB - Congenital acute lymphoblastic leukemia (ALL) is a relatively rare disorder that is characterized by frequent central nervous system involvement that may increase the risk for seizures. Appropriate choice of anticonvulsant therapy with respect to ongoing oncologic treatment is not established in this age group. We report the case of a neonate with ALL who was successfully treated for seizures with levetiracetam monotherapy. This full-term boy did well until 3 days of age when he had an episode of left extremity jerking (07/06/07). Computed tomography of the head demonstrated extensive multifocal intraparenchymal hemorrhages. Initial EEG demonstrated multifocal epileptiform activity. Patient was loaded with Phenobarbital at 20 mg/kg. Complete blood count revealed leukocytosis (78 x 103/mm(3)). Peripheral blood smear contained blastocytes and DNA analysis confirmed B-cell ALL. A second focal seizure was reported on the same day and he was re-loaded with Phenobarbital. Maintenance dosing of Phenobarbital was initiated and no further seizures were noted. A repeat EEG on 7/10/07 remained abnormal with excessive multifocal sharp waves. Continuation of anticonvulsant therapy was recommended. Given concern for interaction between Phenobarbital and planned chemotherapy regimen, oncology requested a non-enzyme inducing anticonvulsant. Phenobarbital was subsequently weaned and Levetiracetam monotherapy initiated at 40 mg/kg/day (07/10/2007). Currently, the patient is seizure free at 8 months of age on Levetiracetam monotherapy. The use of Levetiracetam as monotherapy in neonates has not been formally evaluated and experience is limited. We report the successful use of levetiracetam monotherapy after Phenobarbital load in a neonate with leukemia and localization-related epilepsy. PMID- 19186086 TI - Measuring small compartmental dimensions with low-q angular double-PGSE NMR: The effect of experimental parameters on signal decay. AB - In confined geometries, the MR signal attenuation obtained from single pulsed gradient spin echo (s-PGSE) experiments reflects the dimension of the compartment, and in some cases, its geometry. However, to measure compartment size, high q-values must be applied, requiring high gradient strengths and/or long pulse durations and diffusion times. The angular double PGSE (d-PGSE) experiment has been proposed as a means to extract dimensions of confined geometries using low q-values. In one realization of the d-PGSE experiment, the first gradient pair is fixed along the x-axis, and the orientation of the second gradient pair is varied in the X-Y plane. Such a measurement is sensitive to microscopic anisotropy induced by the boundaries of the restricting compartment, and allows extraction of the compartment dimension. In this study, we have juxtaposed angular d-PGSE experiments and simulations to extract sizes from well characterized NMR phantoms consisting of water filled microcapillaries. We are able to accurately extract sizes of small compartments (5mum) using the angular d PGSE experiment even when the short gradient pulse (SGP) approximation is violated and over a range of mixing and diffusion times. We conclude that the angular d-PGSE experiment may fill an important niche in characterizing compartment sizes in which restricted diffusion occurs. PMID- 19186087 TI - Acupuncture treatment for feline multifocal intervertebral disc disease. AB - A 14-year-old male neutered domestic shorthair cat was admitted to the Veterinary Medical Center, University of Minnesota for evaluation of severe hind limb ataxia, atrophy and paresis. Diagnosis based on physical examination, neurological assessment and magnetic resonance imaging (MRI) was multifocal intervertebral disc disease (IVDD) with dorsal disc protrusion throughout the thoracic and cranial lumbar spine. The Oriental Medicine (OM) diagnosis (pattern identification) was painful obstruction (Bi) syndrome caused by phlegm-heat accumulation with blood stagnation in the spine. High dose prednisolone therapy (1.25mg/kg PO, once daily) initially did not show any significant improvement in clinical signs. The cat was then treated with several modes of acupuncture treatment including dry needle acupuncture, electro-acupuncture and scalp acupuncture along with Tui-Na (hand manipulation in OM) and physical therapy. Significant improvements in mobility, proprioception and spinal posture were noticed and the cat was able to rise, walk and run 4 months after starting acupuncture treatments. This is the first case report of feline IVDD with multiple sites of disc compression which was successfully treated with several modes of acupuncture treatment. PMID- 19186088 TI - Simulation studies on the effect of absorbers on dose distribution in rotational radiotherapy. AB - The effect of cylindrical protector dimensions, material and distance from the source on the dose distribution in rotational radiotherapy was studied to assess the potential protection possibilities of small-sized radiosensitive structures, such as spinal cord. The dose distributions were evaluated in terms of dose at the protected region and surface dose, ratio of the dose at the protected region to the maximum dose, and dose gradient. High-density materials, such as lead, tungsten, gold and cerrobend, along with new polymer-metal composite ones were used in simulation studies, performed by an in-house developed Monte Carlo Radiotherapy Simulator. To ensure correct modeling of the composite materials, simulated attenuation data were verified against experimentally measured data. The dependence of the dose at the protected region from the protector diameter and the field size was established. Protectors of higher density and larger diameter provide not only lower dose at the protected region, but also steeper dose gradient and lower ratio of the dose at the protected region to the treatment dose. For the protection of small structures, high-density protectors placed further from the source allow thicker protectors to be used. The surface dose increases insignificantly for the studied protector-surface distances. The results have shown that shielding properties of composite materials are close to those of lead. PMID- 19186089 TI - The role of immunoglobulin for the treatment of Clostridium difficile infection: a systematic review. AB - Clostridium difficile is the most common infectious cause of nosocomial healthcare-associated diarrhea. The increasing prevalence of C difficile, spread in the community, virulence and frequent relapse has created an urgent need to identify new effective treatments for C. difficile infection. Among these, intravenous immunoglobulin (IVIG) is used for cases of severe C. difficile infection. We undertook a systematic review to examine the published literature pertaining to the use of immunoglobulin for C. difficile infection. Four retrospective studies and five case reports that addressed the use of IVIG for the treatment of C. difficile infection were identified. One study on the use of oral immunoglobulin was identified. Although overall there appear to be benefits to using IVIG in recurrent severe disease, the small sample sizes and lack of control groups in three of the four studies do not allow recommendations to be made regarding the use of immunoglobulin in C. difficile infection. Further research is urgently needed to clarify the role of immunoglobulin--intravenous or oral--for the treatment of C. difficile infection. PMID- 19186090 TI - The prevalence and correlates of hepatitis C virus (HCV) infection and HCV-HIV co infection in a community sample of gay and bisexual men. AB - OBJECTIVE: To describe hepatitis C virus (HCV) and HIV prevalence and co infection, and to examine variables associated with infection in a community sample of men who have sex with men (MSM). METHODS: Data were from an anonymous, cross-sectional study (N=5080) in Canada. Men self-completed a questionnaire and provided an optional saliva specimen for HCV and HIV testing. Polytomous logistic regressions identified variables associated with HCV, HIV, and HCV-HIV co infection. RESULTS: The prevalences of HCV, HIV, and HCV-HIV co-infection were 1.9%, 9.0%, and 0.7%, respectively. The greatest contribution to HCV (odds ratio (OR) 23.66, 95% confidence interval (CI) 9.69-57.73) and HCV-HIV co-infection (OR 26.76, 95% CI 7.97-89.80) was injection drug use. Sexual behaviors and proxies were associated with HIV but not HCV infection. CONCLUSIONS: Results suggest there are subgroups of MSM at risk for HCV. While sexual transmission of HCV was not ruled out, the predominant risk was needle sharing. The greater prevalence of HCV among HIV-positive men suggests the need for greater vigilance in the detection of HCV in this group. PMID- 19186091 TI - [Chondroma of the acromion: an unusual localization]. AB - The scapula is an uncommon site for a chondroma. We report the case of a chondroma in the left acromion, which presented as a painful shoulder in a 19 year-old teenager. The diagnosis was suspected on radiographs and CT scan and confirmed by histological findings. PMID- 19186092 TI - [Algodystrophy versus postoperative infection: use of labelled white cell scans in differential diagnosis in the hand; a case report]. AB - The differential diagnosis between complex regional pain syndrome (CRPS) and postoperative infection is sometimes difficult. We discuss such a case following surgical release of a trigger finger. Because of lack of an accurate diagnosis, the patient was subjected to more and more costly investigations and was operated one more (and unnecessary) time. As white blood cell scintigraphy ((99m)technetium hexamethylpropyleneamineoxime leukocyte [(99m)Tc HMPAO leu]) proved negative, it allowed us to identify with high specificity, a non infectious cause for the problem, and to stop over-investigation and inflation of the costs of treatment. However, the use of white blood cell scintigraphy should remain exceptional, because of its complexity and expense. PMID- 19186093 TI - Baropodometric information return device for foot unloading. AB - Numerous studies on the development of new biofeedback device conception have already been undertaken. The devices are used for some methods of compensation for the data loss on sensors set. Patients with loss of protective pain sensation are unable to modify their gait when abnormal and excessive plantar pressure occurs. Repeated pressure can result in ulcer formation at specific points. For this reason, we have developed a baropodometric biofeedback to prevent injuries by informing the subject when local pressure exceeds a determined threshold. The case study of our visual and auditory biofeedback is encouraging. The system provides a warning system that may play a valuable role in preventing injuries or ulceration by changing the walking pattern without generating dangerous redistribution. PMID- 19186094 TI - A finite element analysis of a T12 vertebra in two consecutive examinations to evaluate the progress of osteoporosis. AB - Osteoporosis is a metabolic disease that causes bones to become fragile and be more likely to break. As basic clinical examinations to detect osteoporosis, dual energy X-ray absorptiometry (DXA) and quantitative computer tomography (QCT) are used. In the framework of a typical clinical examination, QCT scans were obtained from the T12 vertebra of an elderly woman and osteoporosis was diagnosed. One year later, new QCT scans were obtained in order to evaluate her clinical condition. Using both sets as primary information, two patient-specific finite element (FE) models were created and analyzed under compressive load. Vertebral bone was treated as orthotropic material and its elastic modulus was set as an indirect function of Hounsfield Units (HU). Commercial software for medical image processing and FE analysis, along with in house codes, were used for the mechanical analysis of the FE models. Alterations in the geometry/shape of the vertebra as well as in the distributions of several mechanical quantities were detected between the two FE models. As far as the volume of the vertebra is concerned, it augmented by a percentage of 9.7% while the volume of the vertebral body alone increased by 5.6%. In all the maximum values of the mechanical quantities a measurable reduction was observed (axial compressive displacement: 37.9%, von Mises stress: 23.8%, von Mises strains: 15.1%) and all the investigated distributions in the second FE model became smoother. Finally, the percentage of volume with von Mises strains greater than 4500 microstrain dropped from 8.9%, in the first examination, to 4.9% in the second one. Clinically, the prescribed medication seems to have reinforced the structural stability of the vertebra as a whole and through external remodeling the shape of the vertebra changed in a way that the majority of its volume was relieved from stresses and strains of high magnitude. PMID- 19186095 TI - SNP identification in crop plants. AB - In many plants, single nucleotide polymorphism (SNP) markers are increasingly becoming the marker system of choice. However, for many crop plants there are surprisingly low numbers of validated SNP markers available although they are needed in large numbers for studies regarding genetic variation, linkage mapping, population structure analysis, association genetics, map-based gene isolation, and plant breeding. This review summarizes the current status of SNP marker development technologies for major crop plants. It will also provide an outlook into the future regarding possible SNP identification approaches in crop plants on the basis of current development in model systems such as Arabidopsis which will become available with the full sequencing of more plant genomes, genome resequencing, and in conjunction with the next-generation sequencing technologies. PMID- 19186096 TI - Genomics of growth traits in forest trees. AB - Growth traits in trees are fundamental components of adaptation in a forest ecosystem and of productivity in planted forests. A number of processes determine tree growth, which are controlled by genetic and epigenetic factors that respond dynamically to environmental signals throughout centuries. Advances in genomics have allowed an increased comprehension of the complex mechanisms of tree growth and adaptation. Yet, the application of genomics to improving forest productivity and sustainability still entails capturing a large proportion of the total genetic variation controlling the component traits. Nonetheless, genetics and genomics are unifying disciplines that will serve well to dissect the variables and mechanisms of tree growth and development. PMID- 19186097 TI - Virulence factor secretion and translocation by Bordetella species. AB - Here we review the Bordetella virulence secretome with an emphasis on factors that translocate into target cells. Recent advances in understanding the functions of adenylate cyclase toxin, a type 1 secretion system (T1SS) substrate, and pertussis toxin, a type IV secretion system (T4SS) substrate, are briefly described and a compilation of additional secretion systems and secreted factors is provided. Particular attention is devoted to the Bsc type III secretion system (T3SS) and controversies surrounding it. Efforts to identify effector proteins, characterize in vitro and in vivo phenotypes, and the potential role of type III secretion during human infections are discussed. PMID- 19186098 TI - Optical spectroscopy of europium 3,5-dinitrosalicylates-Intense red luminophores. AB - It was found, that alkali metal-europium dinitrosalicylates of composition M(3)Eu(3,5-NO(2)-Sal)(3).nH(2)O (M=Li, Na, K, Cs) are intense red luminophores with wide excitation band. Using methods of optical spectroscopy we studied the influence of nitrogroups and alkali metal counterions on Eu(3+) luminescence efficiency and on processes of excitation energy transfer to Eu(3+) ion in compounds synthesized. The Eu(3+) luminescence and Eu(3+) luminescence excitation spectra, as well as vibrational IR and Raman spectra were investigated. Details of the structure of compounds were discussed. The network of hydrogen bonds in lanthanide dinitrosalicylates is weakening at introduction of large alkali metal ions in compounds and at the increase of the temperature. As a consequence, the long-wavelength shift of the intraligand charge transfer (ILCT) band in Eu(3+) excitation spectra arises at inclusion of Cs(+) cations instead of Li(+) in the crystal lattice of europium dinitrosalicylates and at heating of these compounds. To obtain the energy of the lowest excited triplet state the phosphorescence spectra of alkali metal-gadolinium compounds M(3)Gd(3,5-NO(2)-Sal)(3).nH(2)O, of alkali metal dinitrosalicylate and salicylate salts were measured with time delay. Change of the energies of ligand electronic states and ligand-metal charge transfer state (LM CTS) can give a two-three orders of magnitude enhancement of the Eu(3+) luminescence efficiency in dinitrosalicylates in comparison with salicylates and ten-fold enhancement at the substitution of Li(+) and Na(+) for Cs(+) in dinitrosalicylates. PMID- 19186099 TI - Vibrational spectra of Zn(II) complexes of the amino acids with hydrophobic residues. AB - The infrared and Raman spectra of the bis-chelated Zn(II) complexes of the amino acids glycine, alanine, valine, leucine, isoleucine and phenylalanine were recorded and analyzed in relation to its structural peculiarities. Some comparisons between the recorded spectra are also presented and the characteristics of the carboxylate motions as well as those of the metal-to ligand vibrations are discussed in detail. PMID- 19186101 TI - Serological reports of human infections of H7 and H9 avian influenza viruses in northern China. AB - BACKGROUND: H7 and H9 subtype avian influenza viruses pose a similar threat to humans as H5 virus. OBJECTIVES: This study aims to identify the potential existence of H7 and H9 avian influenza infections in farmers and in poultry workers in northern China regions with highly pathogenic avian influenza (HPAI) H5N1 outbreaks. STUDY DESIGN: Sera were collected from farmers in Xinjiang Uygur autonomous region and Liaoning province and poultry workers in Shandong province. Sera from healthy residents in Shanxi province were used as the controls. H7 and H9 virus infections were examined by hemagglutination inhibition (HI) assay using horse erythrocytes. The titer equal to or greater than 1:160 was considered positive. RESULTS: A total of 583 sera collected from farmers in Xinjiang were tested, and 10 (1.7%) were positive for H9 virus infection. Out of 200 sera collected from Liaoning, two (1.0%) were infected by H9 virus. No H7 virus infection was detected in the above serum samples. Neither H7 nor H9 virus infection was identified in 277 poultry workers of Shandong and in 407 residents of Shanxi. CONCLUSIONS: Although H9 virus infection was limited in farmers from Xinjiang and Liaoning, a public health alert is needed as novel pandemic influenza strains may develop unnoticed given the presence of subclinical infections, and the possibility of re-assortment with prevailing H5N1 virus in these regions. PMID- 19186100 TI - A 10-year molecular survey of herpes simplex virus type 1 in Germany demonstrates a stable and high prevalence of genotypes A and B. AB - BACKGROUND: Recently three different herpes simplex virus type 1 (HSV-1) genotypes (A, B and C) were described on DNA sequence data of the glycoprotein E (gE), G (gG) and I (gI) genes. OBJECTIVE: To type the circulating HSV-1 wild-type strains in Germany and to monitor potential changes in the molecular epidemiology over the past 10 years. STUDY DESIGN: A total of 569 clinical HSV-1 isolates from a 10-year survey in Germany were genotyped by a PCR-based restriction fragment length polymorphism analysis of gG and gI. Recombination analysis of the gE gene sequences was performed to reveal intragenic recombinants. RESULTS: Genotypes A and B strains represented 76% of all strains analyzed and showed a stable distribution within all age groups investigated, independently from the gender. Intergenic gG/gI and intragenic gE recombinants were demonstrated to be less prevalent. Interestingly, for one HIV infected patient a gG/gI genotype switch from A/A to C/A was observed within 3 years. CONCLUSION: The first molecular survey of clinical HSV-1 isolates from Germany demonstrated a stable distribution of two different genotypes and recombinants within age groups and over a period of 10 years. Moreover, homologous recombination seems to be an important feature in the evolution of the HSV-1 genome. PMID- 19186102 TI - Prevalence of undiagnosed obstructive sleep apnea among adult surgical patients in an academic medical center. AB - BACKGROUND: Obstructive sleep apnea (OSA) affects approximately 20% of US adults, of whom about 90% are undiagnosed. While OSA may increase risk of perioperative complications, its prevalence among surgical patients is unknown. We tested the feasibility of screening surgical patients for OSA and determined the prevalence of undiagnosed OSA. METHODS: In a prospective, observational study adult surgical patients were screened for OSA in an academic hospital. Patients without an OSA diagnosis who screened high-risk were offered a home sleep study to determine if they had OSA. The results were compared with polysomnography (PSG) when available. Charts of high-risk patients were examined for postoperative complications. High-risk patients received targeted interventions as part of a hospital safety initiative. RESULTS: There were 2877 patients screened; 661 (23.7%) screened high-risk for OSA, of whom 534 (81%) did not have diagnosed OSA. The portable sleep study detected OSA in 170/207 (82%) high-risk patients without diagnosed OSA. Twenty-six PSGs confirmed OSA in 19 of these patients. Postoperatively there were no respiratory arrests, two unanticipated ICU admissions, and five documented respiratory complications. CONCLUSION: Undiagnosed OSA is prevalent in adult surgical patients. Implementing universal screening is feasible and can identify undiagnosed OSA in many surgical patients. Further investigation is needed into perioperative complications and their prevention for patients with undiagnosed OSA. PMID- 19186103 TI - Associations between sleep disturbance and mental health status: a longitudinal study of Japanese junior high school students. AB - BACKGROUND: A limited number of longitudinal studies have addressed the association between sleep disturbance and mental health status among adolescents. To examine whether each of these is a risk factor for the onset of the other, we conducted a prospective longitudinal study of Japanese adolescents. METHODS: In 2004, we performed a baseline study of students attending three private junior high schools in Tokyo, and in 2006, a follow-up study was performed on the same population. The mean age of the subjects was 13 years. The Pittsburgh Sleep Quality Index was used to evaluate sleep disturbance, and the 12-item General Health Questionnaire was used to evaluate mental health status. RESULTS: The subjects were 698 students, of whom 516 were suitable for analysis. The incidence of newly developed poor mental health status during the 2 years leading to the follow-up study was 35.1%. New onset of poor mental health status was significantly associated with new onset of sleep disturbance and lasting sleep disturbance. The incidence of sleep disturbance during the 2 years leading to the follow-up study was 33.3%. New onset of sleep disturbance was significantly associated with new onset of poor mental health status and lasting poor mental health status. CONCLUSIONS: Sleep disturbance and poor mental health status increase each other's onset risk. PMID- 19186104 TI - XDR tuberculosis and the Indian-subcontinent: effective prevention strategies needed. AB - The Indian-subcontinent is becoming, what could be, the next big battle ground for the control of Extensively Drug Resistant Tuberculosis (XDR-TB). The region caters to not only an ever-increasing incidence of new TB cases, but also a huge burden of already existing TB cases, thereby facing a dual threat for spread of XDR-TB. Most of the health-care facilities are devoid of essential laboratory based infra-structure which is essential for diagnosing XDR-TB. Coupled to this is the lack of knowledge regarding TB diagnosis and treatment amongst the General Practitioners of India and Pakistan, who still treat the bulk of the TB cases. The element of AIDS is also slowly creeping into the equation. Questions have already been raised about the effectiveness of the governmental responses to curb the TB threat. What is needed right away is an effective strategy, incorporating a broad-tiered approach between the government and social sector. This can facilitate greatly in preventing further spread of these resistant strains. PMID- 19186106 TI - Further data on autologous haemopoietic stem cell transplantation in multiple sclerosis. PMID- 19186105 TI - Autologous non-myeloablative haemopoietic stem cell transplantation in relapsing remitting multiple sclerosis: a phase I/II study. AB - BACKGROUND: Autologous non-myeloablative haemopoietic stem cell transplantation is a method to deliver intense immune suppression. We evaluated the safety and clinical outcome of autologous non-myeloablative haemopoietic stem cell transplantation in patients with relapsing-remitting multiple sclerosis (MS) who had not responded to treatment with interferon beta. METHODS: Eligible patients had relapsing-remitting MS, attended Northwestern Memorial Hospital, and despite treatment with interferon beta had had two corticosteroid-treated relapses within the previous 12 months, or one relapse and gadolinium-enhancing lesions seen on MRI and separate from the relapse. Peripheral blood haemopoietic stem cells were mobilised with 2 g per m2 cyclophosphamide and 10 microg per kg per day filgrastim. The conditioning regimen for the haemopoietic stem cells was 200 mg per kg cyclophosphamide and either 20 mg alemtuzumab or 6 mg per kg rabbit antithymocyte globulin. Primary outcomes were progression-free survival and reversal of neurological disability at 3 years post-transplantation. We also sought to investigate the safety and tolerability of autologous non-myeloablative haemopoietic stem cell transplantation. FINDINGS: Between January, 2003, and February, 2005, 21 patients were treated. Engraftment of white blood cells and platelets was on median day 9 (range day 8-11) and patients were discharged from hospital on mean day 11 (range day 8-13). One patient had diarrhoea due to Clostridium difficile and two patients had dermatomal zoster. Two of the 17 patients receiving alemtuzumab developed late immune thrombocytopenic purpura that remitted with standard therapy. 17 of 21 patients (81%) improved by at least 1 point on the Kurtzke expanded disability status scale (EDSS), and five patients (24%) relapsed but achieved remission after further immunosuppression. After a mean of 37 months (range 24-48 months), all patients were free from progression (no deterioration in EDSS score), and 16 were free of relapses. Significant improvements were noted in neurological disability, as determined by EDSS score (p<0.0001), neurological rating scale score (p=0.0001), paced auditory serial addition test (p=0.014), 25-foot walk (p<0.0001), and quality of life, as measured with the short form-36 (SF-36) questionnaire (p<0.0001). INTERPRETATION: Non-myeloablative autologous haemopoietic stem cell transplantation in patients with relapsing-remitting MS reverses neurological deficits, but these results need to be confirmed in a randomised trial. PMID- 19186107 TI - The effect of music intervention in relation to gender during coronary angiographic procedures: a randomized clinical trial. AB - Several studies have evaluated music interventions prior and after coronary angiography and percutaneous coronary intervention (PCI), but there is no clear evidence showing that music has an effect on patients during these procedures. The purpose was to investigate the effects of music on anxiety, angina, pain, relaxation, and comfort in patients during angiographic procedures and to evaluate gender differences. The study was a four-armed, prospective randomized controlled trial included 240 patients undergoing coronary angiography and/or PCI. Patients were allocated to receive relaxing music, MusiCure or standard care during the procedure. Outcome measures were; puncture pain and the discomfort related to it, angina and the discomfort related to it, anxiety, experience of the sound environment, discomfort of lying still, and the doses of anxiolytics and analgesics during the procedure. No differences were found between the music and control groups regarding any of the trial endpoints or gender-related differences. The overall rating of the sound environment and feeling of relaxation was high. In conclusion, music intervention in patients undergoing angiographic procedures was highly feasible, but not effective in this study though the delivery of music went smoothly and did not disturb the examination and patients and staff alike looked favorably on it. PMID- 19186109 TI - Obstructive complications of lichen sclerosus. AB - OBJECTIVE: Young males with lichen sclerosus (LS) commonly present with phimosis; complete urinary obstruction has also been described in this population, but is much rarer. We present the case report of a boy with acute renal failure secondary to urethral obstruction caused by LS, with a review of the literature regarding the occurrence of complete urinary obstruction and renal failure associated with this disease process. MATERIALS AND METHODS: A comprehensive review of the literature was conducted using MEDLINE and EMBASE to characterize the association of lichen sclerosus with significant obstructive complications of the urinary tract. RESULTS: The true incidence of LS is unknown. Reported rates have ranged from 10% to 95% of boys presenting with phimosis. Our review identified 14 cases of complicated urinary obstruction (seven in children) and two cases of renal failure secondary to lichen sclerosus (both in boys). CONCLUSIONS: Distinctive features make this inflammatory process identifiable to the astute physician, such that new onset of phimosis in a school-age boy should raise clinical suspicion of the diagnosis. Successful outcomes, with medical and/or surgical therapy, are commonplace in this cohort. If undiagnosed, however, progression of LS can lead to significant morbidity in the form of renal failure. PMID- 19186108 TI - A virtual screening study of the acetylcholine binding protein using a relaxed complex approach. AB - The nicotinic acetylcholine receptor (nAChR) is a member of the ligand-gated ion channel family and is implicated in many neurological events. Yet, the receptor is difficult to target without high-resolution structures. In contrast, the structure of the acetylcholine binding protein (AChBP) has been solved to high resolution, and it serves as a surrogate structure of the extra-cellular domain in nAChR. Here we conduct a virtual screening study of the AChBP using the relaxed-complex method, which involves a combination of molecular dynamics simulations (to achieve receptor structures) and ligand docking. The library screened through comes from the National Cancer Institute, and its ligands show great potential for binding AChBP in various manners. These ligands mimic the known binders of AChBP; a significant subset docks well against all species of the protein and some distinguish between the various structures. These novel ligands could serve as potential pharmaceuticals in the AChBP/nAChR systems. PMID- 19186110 TI - Is there an ideal age for hypospadias repair? A pilot study. AB - OBJECTIVE: To assess whether repairing hypospadias before or after 18 months affects psychological adjustment, health-related quality of life (HRQoL) and surgical outcome. PATIENTS AND METHODS: Seventy-seven boys aged 6-17 years were assigned to one of two groups, according to whether they had a hypospadias repair before or after the age of 18 months. The surgical outcome was assessed using the pediatric penile perception score by non-involved urologists. A psychologist interviewed the patients to assess HRQoL and gender-role behavior. The child's psychological adjustment was assessed with a questionnaire for parents. RESULTS: The surgical outcome and complication rate were not significantly different between groups. A covariance analysis of HRQoL, gender-role behavior and psychological adjustment as a function of age at the last operation with current age as covariant was performed, but differences did not reach significance. CONCLUSION: This study does not provide evidence to support recommendations concerning the ideal age for hypospadias repair. In the absence of evidence of a benefit of early surgery, anesthesia-related risk factors must be considered when operating in very early infancy. Large, prospective studies, measuring surgical and psychological outcome with similar instruments to those presented may reveal whether there is a true ideal age for hypospadias surgery. PMID- 19186111 TI - Immunodiagnosis of human neurocysticercosis using a synthetic peptide selected by phage-display. AB - The usefulness of a synthetic peptide in the serodiagnosis of Taenia solium human neurocysticercosis (NC) has been evaluated. Phage-displayed peptides were screened with human antibodies to scolex protein antigen from cysticercus cellulosae (SPACc). One clone was found to interact specifically with anti-SPACc IgGs. The corresponding synthetic peptide was found to be recognized in ELISA by NC patient's sera. The study was carried out with sera from 28 confirmed NC patients, 13 control sera and 73 sera from patients suffering from other infectious diseases. A 93% sensibility and a 94.3% specificity was achieved. Figures of 89% and 31.4% of sensibility and specificity were obtained in a SPACc based ELISA. Immunoblotting of SPACc with anti-peptide antibodies revealed a single band of approximately 45 kDa in 1D and four 45 kDa isoforms in 2D-gel electrophoresis. A strong and specific immunostaining in the fibers beneath the suckers, at the base of the rostellum, and in the tissue surrounding the scolex of cysticerci was observed by immunomicroscopy. Our results show that a peptide based immunodiagnostic of neurocisticercosis can be envisioned. PMID- 19186112 TI - 3-Mercaptopropylphosphonic acid modified gold electrode for electrochemical detection of dopamine. AB - It is reported for the first time that the phosphonic acid terminated self assembled layers assembled on a gold surface could be used as a functional interface to sensitively and selectively detect dopamine (DA) in the presence of highly concentrated ascorbic acid (AA). X-ray photoelectron spectroscopic (XPS) and reductive desorption measurements proved that the 3-mercaptopropylphosphonic acid [HS-(CH(2))(3)-PO(3)H(2), MPPA] was successfully immobilized on the gold surface via S-Au bond. Making use of the electrostatic interactions between the negatively charged phosphonic acid groups on the electrode surface and DA and AA, the MPPA modified electrode was successfully used to electrochemically discriminate between DA and AA. Cyclic voltammetry measurements shown that the MPPA modified electrodes almost completely inhibited the electrochemical reaction of AA. Therefore, DA can be selectively and sensitively determined by using the differential pulse voltammetry technique although high concentration of AA is present. PMID- 19186113 TI - Low concentration of extracellular hemoglobin affects shape of RBC in low ion strength sucrose solution. AB - When red blood cells (RBCs) are placed in isotonic sucrose solution with low chloride content dynamic shape response is observed. Generally, it consists of three phases: i) fast spherulation (phase 1), ii) restoration of shape close to discoid one (phase 2), followed by a second slower spherulation in stomatocyte direction (phase 3). This morphological transformation (morphological response (MR)) is rather a fast process, usually comprises 1-2 min interval with disc retransformation time within 20-30 s. We have found that low concentration of extracellular hemoglobin (Hb) in the range 14-44 nM derived from cell lysate significantly accelerated MR at any stage of process. Increasing NaCl concentration up to 10 mM completely eliminated any influence of Hb. Other inhibitors EDTA and chlorpromazine while retarding some phases of MR did not prevent its activation by Hb. It was established that at conditions of low ionic strength Hb strongly binds to native RBC membrane. The number of binding sites was found to be 1-1.6x10(6) molecules per cell and the equilibrium dissociation constant was less than 2x10(-8) M. After heat denaturation of Hb at 70-72 degrees C an ability of Hb to affect MR was lost. Our data provide evidence that extracellular Hb could be involved in shape regulation of red blood cells at least in low ionic strength solutions. PMID- 19186114 TI - Peri-operative complications of video-assisted thoracoscopic surgery (VATS). AB - Video-assisted thoracoscopic surgery (VATS) has multiple indications for diagnosis and treatment of many different thoracic diseases; the commonest are lung wedge resection, pleural and mediastinal biopsy, treatment of pneumothorax, and pleurectomy. Moreover, in recent years a few surgeons have performed routinely major lung anatomic resections by VATS approach, including segmentectomy, lobectomy and pneumonectomy. In our experience VATS constitutes about one-third of all thoracic surgical procedures. In the reviewed literature as in the most frequent complications after VATS procedures are: prolonged air leak, bleeding, infection, postoperative pain, port site recurrence and the need to convert the access in thoracotomy. The complication and mortality rates are generally very low and VATS procedures are considered safe and effective. It is recommended that all thoracic surgery departments audit their VATS procedures for peri-operative morbidity and mortality to compare results and outcomes. PMID- 19186115 TI - Inguinal hernia: state of the art. AB - A review of the history of inguinal hernia repair from the far surgical approach performed by Celso, trought the physiological reconstruction of inguinal canal by Bassini and the introduction of the concept of tensionfree repair, to the newest find in this specialist surgery. Nowadays in addition to the choice of approach (open vs laparoscopic, anterior vs preperitoneal), the plane where placing the mesh (in front of the trasversalis fascia vs preperitoneal space), and the fixation device (suture vs sutureless vs glue), surgeons can select among a wide range of prosthesis. Choosing the proper biomaterial can determine the success of an operation and prevent biomaterial-related complications. Indepth knowledge and understanding of the physical properties of the prosthesis, porosity, and pore size in particular are required. Modern advances in hernia repair are credited with reduced recurrence rate, so surgeons' attention is shifted from preventing recurrence to the new topic of chronic pain after surgery. PMID- 19186116 TI - Microwave tumors ablation: principles, clinical applications and review of preliminary experiences. AB - Local ablative techniques have been developed to enable local control of unresectable tumors. Ablation has been performed with several modalities including ethanol ablation, laser ablation, cryoablation, and radiofrequency ablation. Microwave technology is a new thermal ablation technique for different types of tumors, providing all the benefits of radiofrequency and substantial advantages. Microwave ablation has been applied to liver, lung, kidney and more rarely to bone, pancreas and adrenal glands. Preliminary works show that microwave ablation may be a viable alternative to other ablation techniques in selected patients. However further studies are necessary to confirm short- and long-term effectiveness of the methods and to compare it with other ablative techniques, especially RF. PMID- 19186117 TI - Not a hair out of place: a neat method of otoplasty preparation. PMID- 19186118 TI - Native topology of the designed protein Top7 is not conducive to cooperative folding. AB - Many single-domain proteins with <100 residues fold cooperatively; but the recently designed 92-residue Top7 protein exhibits clearly non-two-state behaviors. In apparent agreement with experiment, we found that coarse-grained, native-centric chain models, including potentials with and without elementary desolvation barriers, predicted that Top7 has a stable intermediate state in which the C-terminal fragment is folded while the rest of the chain remains disordered. We observed noncooperative folding in Top7 models that incorporated nonnative hydrophobic interactions as well. In contrast, free energy profiles deduced from models with desolvation barriers for a set of thirteen natural proteins with similar chain lengths and secondary structure elements suggested that they fold much more cooperatively than Top7. Buttressed by related studies on smaller natural proteins with chain lengths of approximately 40 residues, our findings argue that the de novo native topology of Top7 likely imposed a significant restriction on the cooperativity achievable by any design for this target structure. PMID- 19186119 TI - Teaching old coefficients new tricks: new insight into the meaning of the osmotic and diffusive permeation coefficients. PMID- 19186120 TI - Nucleosome shape dictates chromatin fiber structure. AB - In addition to being the gateway for all access to the eukaryotic genome, chromatin has in recent years been identified as carrying an epigenetic code regulating transcriptional activity. Though much is known about the biochemistry of this code, little is understood regarding the different fiber structures through which the regulation is mediated. Over the last three decades many fiber models have been suggested, but none are able to predict even the basic characteristics of the fiber. In this work, we characterize the set of all possible dense fibers, which includes, but is not limited to, all previously suggested structures. To guide future experimental efforts, we show which fiber characteristics depend on the underlying structure and, crucially, which do not. Addressing the predictive power of these models, we suggest a simple geometric criterion based on the nucleosome shape alone. This enables us to predict the observed characteristics of the condensed chromatin fiber, and how these change with varying nucleosome repeat length. Our approach sheds light on how the in vivo observed heterogeneity in linker lengths can be accommodated within the 30 nm fiber, and suggest an important role for nucleosome surface interactions in the regulation of chromatin structure and function. PMID- 19186121 TI - Interaction between amyloid-beta (1-42) peptide and phospholipid bilayers: a molecular dynamics study. AB - The amyloid-beta (Abeta) peptide is a key aggregate species in Alzheimer's disease. Although important aspects of Abeta peptide aggregation are understood, the initial stage of aggregation from monomer to oligomer is still not clear. One potential mediator of this early aggregation process is interactions of Abeta with anionic cell membranes. We used unconstrained and umbrella sampling molecular dynamics simulations to investigate interactions between the 42-amino acid Abeta peptide and model bilayers of zwitterionic dipalmitoylphosphatidylcholine (DPPC) lipids and anionic dioleoylphosphatidylserine (DOPS) lipids. Using these methods, we determined that Abeta is attracted to the surface of DPPC and DOPS bilayers over the small length scales used in these simulations. We also found supporting evidence that the charge on both the bilayer surface and the peptide affects the free energy of binding of the peptide to the bilayer surface and the distribution of the peptide on the bilayer surface. Our work demonstrates that interactions between the Abeta peptide and lipid bilayer promotes a peptide distribution on the bilayer surface that is prone to peptide-peptide interactions, which can influence the propensity of Abeta to aggregate into higher-order structures. PMID- 19186122 TI - Mechanisms of transition from normal to reentrant electrical activity in a model of rabbit atrial tissue: interaction of tissue heterogeneity and anisotropy. AB - Experimental evidence suggests that regional differences in action potential (AP) morphology can provide a substrate for initiation and maintenance of reentrant arrhythmias in the right atrium (RA), but the relationships between the complex electrophysiological and anatomical organization of the RA and the genesis of reentry are unclear. In this study, a biophysically detailed three-dimensional computer model of the right atrial tissue was constructed to study the role of tissue heterogeneity and anisotropy in arrhythmogenesis. The model of Lindblad et al. for a rabbit atrial cell was modified to incorporate experimental data on regional differences in several ionic currents (primarily, I(Na), I(CaL), I(K1), I(to), and I(sus)) between the crista terminalis and pectinate muscle cells. The modified model was validated by its ability to reproduce the AP properties measured experimentally. The anatomical model of the rabbit RA (including tissue geometry and fiber orientation) was based on a recent histological reconstruction. Simulations with the resultant electrophysiologically and anatomically detailed three-dimensional model show that complex organization of the RA tissue causes breakdown of regular AP conduction patterns at high pacing rates (>11.75 Hz): as the AP in the crista terminalis cells is longer, and electrotonic coupling transverse to fibers of the crista terminalis is weak, high frequency pacing at the border between the crista terminalis and pectinate muscles results in a unidirectional conduction block toward the crista terminalis and generation of reentry. Contributions of the tissue heterogeneity and anisotropy to reentry initiation mechanisms are quantified by measuring action potential duration (APD) gradients at the border between the crista terminalis and pectinate muscles: the APD gradients are high in areas where both heterogeneity and anisotropy are high, such that intrinsic APD differences are not diminished by electrotonic interactions. Thus, our detailed computer model reconstructs complex electrical activity in the RA, and provides new insights into the mechanisms of transition from focal atrial tachycardia into reentry. PMID- 19186123 TI - Diffusion in a fluid membrane with a flexible cortical cytoskeleton. AB - We calculate the influence of a flexible network of long-chain proteins, which is anchored to a fluid membrane, on protein diffusion in this membrane. This is a model for the cortical cytoskeleton and the lipid bilayer of the red blood cell, which we apply to predict the influence of the cytoskeleton on the diffusion coefficient of a mobile band 3 protein. Using the pressure field that the cytoskeleton exerts on the membrane, from the steric repulsion between the diffusing protein and the cytoskeletal filaments, we define a potential landscape for the diffusion within the bilayer. We study the changes to the diffusion coefficient on removal of one type of anchor proteins, e.g., in several hemolytic anemias, as well as for isotropic and anisotropic stretching of the cytoskeleton. We predict an overall increase of the diffusion for a smaller number of anchor proteins and increased diffusion for anisotropic stretching in the direction of the stretch, because of the decrease in the spatial frequency as well as in the height of the potential barriers. PMID- 19186124 TI - Sequence-specific size, structure, and stability of tight protein knots. AB - Approximately 1% of known protein structures display knotted configurations in their native fold, but the function of these configurations is not understood. It has been speculated that the entanglement may inhibit mechanical protein unfolding or transport, e.g., as in cellular threading or translocation processes through narrow biological pores. Protein knot manipulation, e.g., knot tightening and localization, has become possible in single-molecule experiments. Here, we investigate tight peptide knot (TPK) characteristics in detail by pulling selected 3(1) and 4(1)-knotted peptides using all-atom molecular dynamics computer simulations. We find that the 3(1)- and 4(1)-TPK lengths are typically Deltal approximately 47+/- 4 A and 69 +/- 4 A, respectively, for a wide range of tensions (0.1 nN less, similarF less, similar 1.5 nN). The 4(1)-knot length is in agreement with recent atomic force microscopy pulling experiments. Calculated TPK radii of gyration point to a pore diameter of approximately 20 A, below which a translocated knotted protein might get stuck. TPK characteristics, however, may be sequence-specific: we find a different size and structural behavior in polyglycines, and, strikingly, a strong hydrogen bonding and water trapping capability of hydrophobic TPKs. Water capture and release is found to be controllable by the tightening force in a few cases. These mechanisms result in a sequence-specific "locking" and metastability of TPKs, which might lead to a blocking of knotted peptide transport at designated sequence positions. We observe that macroscopic tight 4(1)-knot structures are reproduced microscopically ("figure of eight" versus the "pretzel") and can be tuned by sequence, in contrast to mathematical predictions. Our findings may explain a function of knots in native proteins, challenge previous studies on macromolecular knots, and prove useful in bio- and nanotechnology. PMID- 19186125 TI - Quantifying neurite growth mediated by interactions among secretory vesicles, microtubules, and actin networks. AB - Neurite growth is a fundamental process of neuronal development, which requires both membrane expansions by exocytosis and cytoskeletal dynamics. However, the specific contribution of these processes has not been yet assessed quantitatively. To study and quantify the growth process, we construct a biophysical model in which we relate the overall neurite outgrowth rate to the vesicle dynamics. By considering the complex motion of vesicles in the cell soma, we demonstrate from biophysical consideration that the main step of finding the neurite initiation site relies mainly on a two-dimensional diffusion/sequestration/fusion at the cell surface and we obtain a novel formula for the flux of vesicles at the neurite base. In the absence of microtubules, we show that a nascent neurite initiated by vesicular delivery can only reach a small length. By adding the microtubule dynamics to the secretory pathway and using stochastic analysis and simulations, we study the complex dynamics of neurite growth. Within this model, depending on the coupling parameter between the microtubules and the neurite, we find different regimes of growth, which describe dendritic and axonal growth. To validate one aspect of our model, we demonstrate that the experimental flux of TI-VAMP but not Synaptobrevin 2 vesicles contributes to the neurite growth. We conclude that although vesicles can be generated randomly in the cell body, the search for the neurite position using the microtubule network and diffusion is quite fast. Furthermore, when the TI-VAMP vesicular flow is large enough, the interactions between the microtubule bundle and the neurite control the growth process. In addition, all of these processes intimately cooperate to mediate the various modes of neurite growth: the model predicts three different growing modes including, in addition to the stable axonal growth and the stochastic dendritic growth, a fast oscillatory regime. Finally our study demonstrates that cytoskeletal dynamics is necessary to generate long protrusion, while vesicular delivery alone can only generate small neurite. PMID- 19186126 TI - Computational modeling of structurally conserved cancer mutations in the RET and MET kinases: the impact on protein structure, dynamics, and stability. AB - Structural and biochemical characterization of protein kinases that confer oncogene addiction and harbor a large number of disease-associated mutations, including RET and MET kinases, have provided insights into molecular mechanisms associated with the protein kinase activation in human cancer. In this article, structural modeling, molecular dynamics, and free energy simulations of a structurally conserved mutational hotspot, shared by M918T in RET and M1250T in MET kinases, are undertaken to quantify the molecular mechanism of activation and the functional role of cancer mutations in altering protein kinase structure, dynamics, and stability. The mechanistic basis of the activating RET and MET cancer mutations may be driven by an appreciable free energy destabilization of the inactive kinase state in the mutational forms. According to our results, the locally enhanced mobility of the cancer mutants and a higher conformational entropy are counterbalanced by a larger enthalpy loss and result in the decreased thermodynamic stability. The computed protein stability differences between the wild-type and cancer kinase mutants are consistent with circular dichroism spectroscopy and differential scanning calorimetry experiments. These results support the molecular mechanism of activation, which causes a detrimental imbalance in the dynamic equilibrium shifted toward the active form of the enzyme. Furthermore, computer simulations of the inhibitor binding with the oncogenic and drug-resistant RET mutations have also provided a plausible molecular rationale for the observed differences in the inhibition profiles, which is consistent with the experimental data. Finally, structural mapping of RET and MET cancer mutations and the computed protein stability changes suggest a similar mechanism of activation, whereby the cancer mutations which display the higher oncogenic activity tend to have the greatest destabilization effect on the inactive kinase structure. PMID- 19186127 TI - What determines the structure and stability of KFFE monomers, dimers, and protofibrils? AB - The self-assembly of the KFFE peptide was studied using replica exchange molecular dynamics simulations with a fully atomic description of the peptide and explicit solvent. The relative roles of the aromatic residues and oppositely charged end groups in stabilizing the earliest oligomers and the end-products of aggregation were investigated. beta and non-beta-peptide conformations compete in the monomeric state as a result of a balancing between the high beta-sheet propensity of the phenylalanine residues and charge-charge interactions that favor non-beta-conformations. Dimers are present in beta- and non-beta-sheet conformations and are stabilized primarily by direct and water-mediated charge charge interactions between oppositely charged side chains and between oppositely charged termini, with forces between aromatic residues playing a minor role. Dimerization to a beta-sheet, fibril-competent state, is seen to be a cooperative process, with the association process inducing beta-structure in otherwise non beta-monomers. We propose a model for the KFFE fibril, with mixed interface and antiparallel sheet and strand arrangements, which is consistent with experimental electron microscopy measurements. Both aromatic and charge-charge interactions contribute to the fibril stability, although the dominant contribution arises from electrostatic interactions. PMID- 19186128 TI - Comparison of deterministic and stochastic models of the lac operon genetic network. AB - The lac operon has been a paradigm for genetic regulation with positive feedback, and several modeling studies have described its dynamics at various levels of detail. However, it has not yet been analyzed how stochasticity can enrich the system's behavior, creating effects that are not observed in the deterministic case. To address this problem we use a comparative approach. We develop a reaction network for the dynamics of the lac operon genetic switch and derive corresponding deterministic and stochastic models that incorporate biological details. We then analyze the effects of key biomolecular mechanisms, such as promoter strength and binding affinities, on the behavior of the models. No assumptions or approximations are made when building the models other than those utilized in the reaction network. Thus, we are able to carry out a meaningful comparison between the predictions of the two models to demonstrate genuine effects of stochasticity. Such a comparison reveals that in the presence of stochasticity, certain biomolecular mechanisms can profoundly influence the region where the system exhibits bistability, a key characteristic of the lac operon dynamics. For these cases, the temporal asymptotic behavior of the deterministic model remains unchanged, indicating a role of stochasticity in modulating the behavior of the system. PMID- 19186129 TI - Curvature-driven pore growth in charged membranes during charge-pulse and voltage clamp experiments. AB - We find that curvature-driven growth of pores in electrically charged membranes correctly reproduces charge-pulse experiments. Our model, consisting of a Langevin equation for the time dependence of the pore radius coupled to an ordinary differential equation for the number of pores, captures the statistics of the pore population and its effect on the membrane conductance. The calculated pore radius is a linear, and not an exponential, function of time, as observed experimentally. Two other important features of charge-pulse experiments are recovered: pores reseal for low and high voltages but grow irreversibly for intermediate values of the voltage. Our set of coupled ordinary differential equations is equivalent to the partial differential equation used previously to study pore dynamics, but permits the study of longer timescales necessary for the simulations of voltage-clamp experiments. An effective phase diagram for such experiments is obtained. PMID- 19186130 TI - Blind test of physics-based prediction of protein structures. AB - We report here a multiprotein blind test of a computer method to predict native protein structures based solely on an all-atom physics-based force field. We use the AMBER 96 potential function with an implicit (GB/SA) model of solvation, combined with replica-exchange molecular-dynamics simulations. Coarse conformational sampling is performed using the zipping and assembly method (ZAM), an approach that is designed to mimic the putative physical routes of protein folding. ZAM was applied to the folding of six proteins, from 76 to 112 monomers in length, in CASP7, a community-wide blind test of protein structure prediction. Because these predictions have about the same level of accuracy as typical bioinformatics methods, and do not utilize information from databases of known native structures, this work opens up the possibility of predicting the structures of membrane proteins, synthetic peptides, or other foldable polymers, for which there is little prior knowledge of native structures. This approach may also be useful for predicting physical protein folding routes, non-native conformations, and other physical properties from amino acid sequences. PMID- 19186131 TI - Determinants of water permeability through nanoscopic hydrophilic channels. AB - Naturally occurring pores show a variety of polarities and sizes that are presumably directly linked to their biological function. Many biological channels are selective toward permeants similar or smaller in size than water molecules, and therefore their pores operate in the regime of single-file water pores. Intrinsic factors affecting water permeability through such pores include the channel-membrane match, the structural stability of the channel, the channel geometry and channel-water affinity. We present an extensive molecular dynamics study on the role of the channel geometry and polarity on the water osmotic and diffusive permeability coefficients. We show that the polarity of the naturally occurring peptidic channels is close to optimal for water permeation, and that the water mobility for a wide range of channel polarities is essentially length independent. By systematically varying the geometry and polarity of model hydrophilic pores, based on the fold of gramicidin A, the water density, occupancy, and permeability are studied. Our focus is on the characterization of the transition between different permeation regimes in terms of the structure of water in the pores, the average pore occupancy and the dynamics of the permeating water molecules. We show that a general relationship between osmotic and diffusive water permeability coefficients in the single-file regime accounts for the time averaged pore occupancy, and that the dynamics of the permeating water molecules through narrow non single file channels effectively behaves like independent single-file columns. PMID- 19186132 TI - Overexpression of rhodopsin alters the structure and photoresponse of rod photoreceptors. AB - Rhodopsins are densely packed in rod outer-segment membranes to maximize photon absorption, but this arrangement interferes with transducin activation by restricting the mobility of both proteins. We attempted to explore this phenomenon in transgenic mice that overexpressed rhodopsin in their rods. Photon capture was improved, and, for a given number of photoisomerizations, bright flash responses rose more gradually with a reduction in amplification--but not because rhodopsins were more tightly packed in the membrane. Instead, rods increased their outer-segment diameters, accommodating the extra rhodopsins without changing the rhodopsin packing density. Because the expression of other phototransduction proteins did not increase, transducin and its effector phosphodiesterase were distributed over a larger surface area. That feature, as well as an increase in cytosolic volume, was responsible for delaying the onset of the photoresponse and for attenuating its amplification. PMID- 19186134 TI - Constitutive dimerization of the G-protein coupled receptor, neurotensin receptor 1, reconstituted into phospholipid bilayers. AB - Neurotensin receptor 1 (NTS1), a Family A G-protein coupled receptor (GPCR), was expressed in Escherichia coli as a fusion with the fluorescent proteins eCFP or eYFP. A fluorophore-tagged receptor was used to study the multimerization of NTS1 in detergent solution and in brain polar lipid bilayers, using fluorescence resonance energy transfer (FRET). A detergent-solubilized receptor was unable to form FRET-competent complexes at concentrations of up to 200 nM, suggesting that the receptor is monomeric in this environment. When reconstituted into a model membrane system at low receptor density, the observed FRET was independent of agonist binding, suggesting constitutive multimer formation. In competition studies, decreased FRET in the presence of untagged NTS1 excludes the possibility of fluorescent protein-induced interactions. A simulation of the experimental data indicates that NTS1 exists predominantly as a homodimer, rather than as higher-order multimers. These observations suggest that, in common with several other Family A GPCRs, NTS1 forms a constitutive dimer in lipid bilayers, stabilized through receptor-receptor interactions in the absence of other cellular signaling components. Therefore, this work demonstrates that well characterized model membrane systems are useful tools for the study of GPCR multimerization, allowing fine control over system composition and complexity, provided that rigorous control experiments are performed. PMID- 19186133 TI - Predictions suggesting a participation of beta-sheet configuration in the M2 domain of the P2X(7) receptor: a novel conformation? AB - Scanning experiments have shown that the putative TM2 domain of the P2X(7) receptor (P2X(7)R) lines the ionic pore. However, none has identified an alpha helix structure, the paradigmatic secondary structure of ion channels in mammalian cells. In addition, some researchers have suggested a beta-sheet conformation in the TM2 domain of P2X(2). These data led us to investigate a new architecture within the P2X receptor family. P2X(7)R is considered an intriguing receptor because its activation induces nonselective large pore formation, in contrast to the majority of other ionic channel proteins in mammals. This receptor has two states: a low-conductance channel (approximately 10 pS) and a large pore (> 400 pS). To our knowledge, one fundamental question remains unanswered: Are the P2X(7)R channel and the pore itself the same entity or are they different structures? There are no structural data to help solve this question. Thus, we investigated the hydrophobic M2 domain with the aim of predicting the fitted position and the secondary structure of the TM2 segment from human P2X(7)R (hP2X(7)R). We provide evidence for a beta-sheet conformation, using bioinformatics algorithms and molecular-dynamics simulation in conjunction with circular dichroism in different environments and Fourier transform infrared spectroscopy. In summary, our study suggests the possibility that a segment composed of residues from part of the M2 domain and part of the putative TM2 segment of P2X(7)R is partially folded in a beta-sheet conformation, and may play an important role in channel/pore formation associated with P2X(7)R activation. It is important to note that most nonselective large pores have a transmembrane beta-sheet conformation. Thus, this study may lead to a paradigmatic change in the P2X(7)R field and/or raise new questions about this issue. PMID- 19186135 TI - Functional and structural characterization of factor Xa dimer in solution. AB - Previous studies showed that binding of water-soluble phosphatidylserine (C6PS) to bovine factor Xa (FXa) leads to Ca2+-dependent dimerization in solution. We report the effects of Ca2+, C6PS, and dimerization on the activity and structure of human and bovine FXa. Both human and bovine dimers are 10(6)- to 10(7)-fold less active toward prothrombin than the monomer, with the decrease being attributed mainly to a substantial decrease in k(cat). Dimerization appears not to block the active site, since amidolytic activity toward a synthetic substrate is largely unaffected. Circular dichroism reveals a substantial change in tertiary or quaternary structure with a concomitant decrease in alpha-helix upon dimerization. Mass spectrometry identifies a lysine (K(270)) in the catalytic domain that appears to be buried at the dimer interface and is part of a synthetic peptide sequence reported to interfere with factor Va (FVa) binding. C6PS binding exposes K(351) (part of a reported FVa binding region), K(242) (adjacent to the catalytic triad), and K(420) (part of a substrate exosite). We interpret our results to mean that C6PS-induced dimerization produces substantial conformational changes or domain rearrangements such that structural data on PS activated FXa is required to understand the structure of the FXa dimer or the FXa FVa complex. PMID- 19186136 TI - Fold-unfold transitions in the selectivity and mechanism of action of the N terminal fragment of the bactericidal/permeability-increasing protein (rBPI(21)). AB - Septic or endotoxic shock is a common cause of death in hospital intensive care units. In the last decade numerous antimicrobial peptides and proteins have been tested in the search for an efficient drug to treat this lethal disease. Now in phase III clinical trials, rBPI(21), a recombinant N-terminal fragment of the bactericidal/permeability-increasing protein (BPI), is a promising drug to reduce lesions caused by meningococcal sepsis. We correlated structural and stability data with functional information of rBPI(21) bound to both model systems of eukaryotic and bacterial membranes. On interaction with membranes, rBPI(21) loses its conformational stability, as studied by circular dichroism. This interaction of rBPI(21) at membrane level was higher in the presence of negatively charged phospholipid relatively to neutral ones, with higher partition coefficients (K(p)), suggesting a preference for bacterial membranes over mammalian membranes. rBPI(21) binding to membranes is reinforced when its disulfide bond is broken due to conformational changes of the protein. This interaction is followed by liposome aggregation due to unfolding, which ensures protein aggregation, and interfacial localization of rBPI(21) in membranes, as studied by extensive quenching by acrylamide and 5-deoxylstearic acid and not by 16-deoxylstearic acid. An uncommon model of the selectivity and mechanism of action is proposed, where membrane induces unfolding of the antimicrobial protein, rBPI(21). The unfolding ensures protein aggregation, established by protein-protein interaction at membrane surface or between adjacent membranes covered by the unfolded protein. This protein aggregation step may lead to membrane perturbation. PMID- 19186137 TI - Intramembrane water associated with TOAC spin-labeled alamethicin: electron spin echo envelope modulation by D2O. AB - Alamethicin is a 20-residue, hydrophobic, helical peptide, which forms voltage sensitive ion channels in lipid membranes. The helicogenic, nitroxyl amino acid TOAC was substituted isosterically for Aib at residue positions 1, 8, or 16 in a F50/5 alamethicin analog to enable EPR studies. Electron spin-echo envelope modulation (ESEEM) spectroscopy was used to investigate the water exposure of TOAC-alamethicin introduced into membranes of saturated or unsaturated diacyl phosphatidylcholines that were dispersed in D2O. Echo-detected EPR spectra were used to assess the degree of assembly of the peptide in the membrane, via the instantaneous diffusion from intermolecular spin-spin interactions. The profile of residue exposure to water differs between membranes of saturated and unsaturated lipids. In monounsaturated dioleoyl phosphatidylcholine, D2O-ESEEM intensities decrease from TOAC(1) to TOAC(8) and TOAC(16) but not uniformly. This is consistent with a transmembrane orientation for the protoassembled state, in which TOAC(16) is located in the bilayer leaflet opposite to that of TOAC(1) and TOAC(8). Relative to the monomer in fluid bilayers, assembled alamethicin is disposed asymmetrically about the bilayer midplane. In saturated dimyristoyl phosphatidylcholine, the D2O-ESEEM intensity is greatest for TOAC(8), indicating a more superficial location for alamethicin, which correlates with the difference in orientation between gel- and fluid-phase membranes found by conventional EPR of TOAC-alamethicin in aligned phosphatidylcholine bilayers. Increasing alamethicin/lipid ratio in saturated phosphatidylcholine shifts the profile of water exposure toward that with unsaturated lipid, consistent with proposals of a critical concentration for switching between the two different membrane associated states. PMID- 19186138 TI - Biophysical characterization of a new phospholipid analogue with a spin-labeled unsaturated fatty acyl chain. AB - Spin-labeled analogs of phospholipids have been used widely to characterize the biophysical properties of membranes. We describe synthesis and application of a new spin-labeled phospholipid analog, SL-POPC. The advantage of this molecule is that the EPR active doxyl group is linked to an unsaturated fatty acyl chain different to saturated phospholipid analogs used so far. The need for those analogs arises from the fact that biological membranes contain unsaturated phospholipids to a large extent. The biophysical properties of SL-POPC in membranes were characterized using EPR and NMR spectroscopy and compared with those of the saturated spin-labeled phospholipid, SL-PSPC. To this end, POPC membranes were labeled with either analog to assess whether the spin-labeled counterpart SL-POPC mimics the membrane properties better than the often used SL PSPC. The results show that SL-POPC and SL-PSPC explore different molecular environments of the bilayer, and that the type and degree of perturbation of bilayer caused by the label moiety also differs between both analogs. We found that SL-POPC is more appropriate to assess the versatile dynamics of POPC membranes than SL-PSPC. PMID- 19186139 TI - Adsorption of GST-PI3Kgamma at the air-buffer interface and at substrate and nonsubstrate phospholipid monolayers. AB - The recruitment of phosphoinositide 3-kinase gamma (PI3Kgamma) to the cell membrane is a crucial requirement for the initiation of inflammation cascades by second-messenger production. In addition to identifying other regulation pathways, it has been found that PI3Kgamma is able to bind phospholipids directly. In this study, the adsorption behavior of glutathione S-transferase (GST)-PI3Kgamma to nonsubstrate model phospholipids, as well as to commercially available substrate inositol phospholipids (phosphoinositides), was investigated by use of infrared reflection-absorption spectroscopy (IRRAS). The nonsubstrate phospholipid monolayers also yielded important information about structural requirements for protein adsorption. The enzyme did not interact with condensed zwitterionic or anionic monolayers; however, it could penetrate into uncompressed fluid monolayers. Compression to values above its equilibrium pressure led to a squeezing out and desorption of the protein. Protein affinity for the monolayer surface increased considerably when the lipid had an anionic headgroup and contained an arachidonoyl fatty acyl chain in sn-2 position. Similar results on a much higher level were observed with substrate phosphoinositides. No structural response of GST-PI3Kgamma to lipid interaction was detected by IRRAS. On the other hand, protein adsorption caused a condensing effect in phosphoinositide monolayers. In addition, the protein reduced the charge density at the interface probably by shifting the pK values of the phosphate groups attached to the inositol headgroups. Because of their strongly polar headgroups, an interaction of the inositides with the water molecules of the subphase can be expected. This interaction is disturbed by protein adsorption, causing the ionization state of the phosphates to change. PMID- 19186140 TI - Interaction of tea catechin (-)-epigallocatechin gallate with lipid bilayers. AB - A major component of green tea extracts, catechin (-)-Epigallocatechin gallate (EGCg), has been reported to be biologically active and interacting with membranes. A recent study reported drastic effects of EGCg on giant unilamellar vesicles (GUVs). In particular, EGCg above 30 microM caused GUVs to burst. Here we investigated the effect of EGCg on single GUVs at lower concentrations, believing that its molecular mechanism would be more clearly revealed. We used the micropipette aspiration method, by which the changes of surface area and volume of a GUV could be measured as a result of interaction with EGCg. We also used x-ray diffraction to measure the membrane thinning effect by EGCg. To understand the property of EGCg, we compared its effect with other membrane active molecules, including pore-forming peptide magainin, the turmeric (curry) extract curcumin, and detergent Triton X100. We found the effect of EGCg somewhat unique. Although EGCg readily binds to lipid bilayers, its membrane area expansion effect is one order of magnitude smaller than curcumin. EGCg also solubilizes lipid molecules from lipid bilayers without forming pores, but its effect is different from that of Triton X100. PMID- 19186141 TI - Mechanical distortion of single actin filaments induced by external force: detection by fluorescence imaging. AB - Actin is a major component of the cytoskeleton that transmits mechanical stress in both muscle and nonmuscle cells. As the first step toward developing a "bio nano strain gauge" that would be able to report the mechanical stress imposed on an actin filament, we quantitatively examined the fluorescence intensity of dyes attached to single actin filaments under various tensile forces (5-20 pN). Tensile force was applied via two optically trapped plastic beads covalently coated with chemically modified heavy meromyosin molecules that were attached to both end regions of an actin filament. As a result, we found that the fluorescence intensity of an actin filament, where 20% of monomers were labeled with tetramethylrhodamine (TMR)-5-maleimide at Cys(374) and the filamentous structure was stabilized with nonfluorescent phalloidin, decreased by approximately 6% per 10 pN of the applied force, whereas the fluorescence intensity of an actin filament labeled with either BODIPY TMR cadaverin iodoacetamide at Cys(374) or rhodamine-phalloidin showed only an approximately 2% decrease per 10 pN of the applied force. On the other hand, spectroscopic measurements of actin solutions showed that the fluorescence intensity of TMR actin increased 1.65-fold upon polymerization (G-F transformation), whereas that of BODIPY-actin increased only 1.06-fold. These results indicate that the external force distorts the filament structure, such that the microenvironment around Cys(374) approaches that in G-actin. We thus conclude that the fluorescent dye incorporated into an appropriate site of actin can report the mechanical distortion of the binding site, which is a necessary condition for the bio-nano strain gauge. PMID- 19186142 TI - Dynamics of thin-filament activation in rabbit skeletal muscle fibers examined by time-resolved x-ray diffraction. AB - By using skinned-rabbit skeletal muscle fibers, the time courses of changes of thin filament-based x-ray reflections were followed at a 3.4-ms time resolution during thin-filament activation. To discriminate between the effects of calcium binding and myosin binding on thin-filament activity, measurements were performed after caged-calcium photolysis in fibers with full-filament or no-filament overlap, or during force recovery after a quick release. All three reflections examined, i.e., the second actin layer line (second ALL, reporting the tropomyosin movement), the sixth ALL (reporting actin structural change), and the meridional troponin reflections, exhibited calcium-induced and myosin-induced components, but their rate constants and polarities were different. Generally, calcium-induced components exhibited fast rate constants (>100 s(-1)). The myosin induced components of the second ALL had a rate constant similar to that of the force (7-10 s(-1)), but that of the sixth ALL was apparently faster. The myosin induced component of troponin reflection was the only one with negative polarity, and was too slow to be analyzed with this protocol. The results suggest that the three regulation-related proteins change their structures with different rate constants, and the significance of these findings is discussed in the context of a cooperative thin-filament activation mechanism. PMID- 19186143 TI - Small anion with higher valency retards the compaction of DNA in the presence of multivalent cation. AB - It has been established that, upon the addition of multivalent cations, long DNA chains in an aqueous solution exhibit a remarkable discrete transition from a coil state to a compact state at the level of a single chain. In this study, we investigated the polyelectrolyte nature of DNA with the experimental methodology of single-DNA observation, and provide a theoretical interpretation. We examined the effects of co-ions with different valencies (Cl(-), SO4(2-), PO4(3-)) on DNA compaction. As a result, we found that co-ions with a greater valency induce the coil state rather than the compact state. Based on a simple model with mean-field approximation that considered ion pairing, we show how the increase in entropy of small ions contributes to the stability of the compact state, by overcoming entropic penalties such as elastic confinement of the chain and a decrease in the translational freedom of counterions accompanied by charge neutralization. PMID- 19186144 TI - The c2 domains of human synaptotagmin 1 have distinct mechanical properties. AB - Synaptotagmin 1 (Syt1) is the Ca(+2) receptor for fast, synchronous vesicle fusion in neurons. Because membrane fusion is an inherently mechanical, force driven event, Syt1 must be able to adapt to the energetics of the fusion apparatus. Syt1 contains two C2 domains (C2A and C2B) that are homologous in sequence and three-dimensional in structure; yet, a number of observations have suggested that they have distinct biochemical and biological properties. In this study, we analyzed the mechanical stability of the C2A and C2B domains of human Syt1 using single-molecule atomic force microscopy. We found that stretching the C2AB domains of Syt1 resulted in two distinct unfolding force peaks. The larger force peak of approximately 100 pN was identified as C2B and the second peak of approximately 50 pN as C2A. Furthermore, a significant fraction of C2A domains unfolded through a low force intermediate that was not observed in C2B. We conclude that these domains have different mechanical properties. We hypothesize that a relatively small stretching force may be sufficient to deform the effector binding regions of the C2A domain and modulate the affinity for soluble N ethylmaleimide-sensitive factor (NSF) attachment protein receptors (SNAREs), phospholipids, and Ca(+2). PMID- 19186145 TI - Structural differences between Abeta(1-40) intermediate oligomers and fibrils elucidated by proteolytic fragmentation and hydrogen/deuterium exchange. AB - The aggregation of amyloid-beta protein (Abeta) in vivo is a critical pathological event in Alzheimer's disease. Although more and more evidence shows that the intermediate oligomers are the primary neurotoxic species in Alzheimer's disease, the particular structural features responsible for the toxicity of these intermediates are poorly understood. We measured the peptide level solvent accessibility of multiple Abeta(1-40) aggregated states using hydrogen exchange detected by mass spectrometry. A gradual reduction in solvent accessibility, spreading from the C-terminal region to the N-terminal region was observed with ever more aggregated states of Abeta peptide. The observed hydrogen exchange protection begins with reporter peptides 20-34 and 35-40 in low molecular weight oligomers found in fresh samples and culminates with increasing solvent protection of reporter peptide 1-16 in long time aged fibrillar species. The more solvent exposed structure of intermediate oligomers in the N-termini relative to well-developed fibrils provides a novel explanation for the structure-dependent neurotoxicity of soluble oligomers reported previously. PMID- 19186146 TI - Tensile mechanics of alanine-based helical polypeptide: force spectroscopy versus computer simulations. AB - In nature, an alpha-helix is commonly used to build thermodynamically stable and mechanically rigid protein conformations. In view of growing interest in the mechanical rigidity of proteins, we measured the tensile profile of an alanine based alpha-helical polypeptide on an atomic-force microscope to investigate the basic mechanics of helix extension with minimal interference from side-chain interactions. The peptide was extended to its maximum contour length with much less force than in reported cases of poly-L-Glu or poly-L-Lys, indicating that chain stiffness strongly depended on the physicochemical properties of side chains, such as their bulkiness. The low tensile-force extension originated presumably in locally unfolded parts because of spontaneous structural fluctuations. In 50% trifluoroethanol, the well-known helix-promoting agent, the rigidity of the sample polypeptide was markedly increased. Computer simulations of the peptide-stretching process showed that a majority of constituent residues underwent a transition from an alpha-helical to an extended conformation by overcoming an energy barrier around psi approximately 0 degrees on the Ramachandran plot. The observed lability of an isolated helix signified the biological importance of the lateral bundling of helices to maintain a rigid protein structure. PMID- 19186147 TI - Interaction of IAPP and insulin with model interfaces studied using neutron reflectometry. AB - The islet amyloid polypeptide (IAPP) and insulin are coproduced by the beta-cells of the pancreatic islets of Langerhans. Both peptides can interact with negatively charged lipid membranes. The positively charged islet amyloid polypeptide partially inserts into these membranes and subsequently forms amyloid fibrils. The amyloid fibril formation of insulin is also accelerated by the presence of negatively charged lipids, although insulin has a negative net charge at neutral pH-values. We used water-polymer model interfaces to differentiate between the hydrophobic and electrostatic interactions that can drive these peptides to adsorb at an interface. By applying neutron reflectometry, the scattering-length density profiles of IAPP and insulin, as adsorbed at three different water-polymer interfaces, were determined. The islet amyloid polypeptide most strongly adsorbed at a hydrophobic poly-(styrene) surface, whereas at a hydrophilic, negatively charged poly-(styrene sulfonate) interface, the degree of adsorption was reduced by 50%. Almost no IAPP adsorption was evident at this negatively charged interface when we added 100 mM NaCl. On the other hand, negatively charged insulin was most strongly attracted to a hydrophilic, negatively charged interface. Our results suggest that IAPP is strongly attracted to a hydrophobic surface, whereas the few positive charges of IAPP cannot warrant a permanent immobilization of IAPP at a hydrophilic, negatively charged surface at an ionic strength of 100 mM. Furthermore, the interfacial accumulation of insulin at a hydrophilic, negatively charged surface may represent a favorable precondition for nucleus formation and fibril formation. PMID- 19186148 TI - Pulsed EPR determination of water accessibility to spin-labeled amino acid residues in LHCIIb. AB - Membrane proteins reside in a structured environment in which some of their residues are accessible to water, some are in contact with alkyl chains of lipid molecules, and some are buried in the protein. Water accessibility of residues may change during folding or function-related structural dynamics. Several techniques based on the combination of pulsed electron paramagnetic resonance (EPR) with site-directed spin labeling can be used to quantify such water accessibility. Accessibility parameters for different residues in major plant light-harvesting complex IIb are determined by electron spin echo envelope modulation spectroscopy in the presence of deuterated water, deuterium contrast in transversal relaxation rates, analysis of longitudinal relaxation rates, and line shape analysis of electron-spin-echo-detected EPR spectra as well as by the conventional techniques of measuring the maximum hyperfine splitting and progressive saturation in continuous-wave EPR. Systematic comparison of these parameters allows for a more detailed characterization of the environment of the spin-labeled residues. These techniques are applicable independently of protein size and require approximately 10-20 nmol of singly spin-labeled protein per sample. For a residue close to the N-terminus, in a domain unresolved in the existing x-ray structures of light-harvesting complex IIb, all methods indicate high water accessibility. PMID- 19186149 TI - Pressure-induced changes in the structure and function of the kinesin-microtubule complex. AB - Kinesin-1 is an ATP-driven molecular motor that "walks" along a microtubule by working two heads in a "hand-over-hand" fashion. The stepping motion is well coordinated by intermolecular interactions between the kinesin head and microtubule, and is sensitively changed by applied forces. We demonstrate that hydrostatic pressure works as an inhibitory action on kinesin motility. We developed a high-pressure microscope that enables the application of hydrostatic pressures of up to 200 MPa (2000 bar). Under high-pressure conditions, taxol stabilized microtubules were shortened from both ends at the same speed. The sliding velocity of kinesin motors was reversibly changed by pressure, and reached half-maximal value at approximately 100 MPa. The pressure-velocity relationship was very close to the force-velocity relationship of single kinesin molecules, suggesting a similar inhibitory mechanism on kinesin motility. Further analysis showed that the pressure mainly affects the stepping motion, but not the ATP binding reaction. The application of pressure is thought to enhance the structural fluctuation and/or association of water molecules with the exposed regions of the kinesin head and microtubule. These pressure-induced effects could prevent kinesin motors from completing the stepping motion. PMID- 19186150 TI - UVA generates pyrimidine dimers in DNA directly. AB - There is increasing evidence that UVA radiation, which makes up approximately 95% of the solar UV light reaching the Earth's surface and is also commonly used for cosmetic purposes, is genotoxic. However, in contrast to UVC and UVB, the mechanisms by which UVA produces various DNA lesions are still unclear. In addition, the relative amounts of various types of UVA lesions and their mutagenic significance are also a subject of debate. Here, we exploit atomic force microscopy (AFM) imaging of individual DNA molecules, alone and in complexes with a suite of DNA repair enzymes and antibodies, to directly quantify UVA damage and reexamine its basic mechanisms at a single-molecule level. By combining the activity of endonuclease IV and T4 endonuclease V on highly purified and UVA-irradiated pUC18 plasmids, we show by direct AFM imaging that UVA produces a significant amount of abasic sites and cyclobutane pyrimidine dimers (CPDs). However, we find that only approximately 60% of the T4 endonuclease V-sensitive sites, which are commonly counted as CPDs, are true CPDs; the other 40% are abasic sites. Most importantly, our results obtained by AFM imaging of highly purified native and synthetic DNA using T4 endonuclease V, photolyase, and anti-CPD antibodies strongly suggest that CPDs are produced by UVA directly. Thus, our observations contradict the predominant view that as-yet unidentified photosensitizers are required to transfer the energy of UVA to DNA to produce CPDs. Our results may help to resolve the long-standing controversy about the origin of UVA-produced CPDs in DNA. PMID- 19186152 TI - Dynamics of type IV pili is controlled by switching between multiple states. AB - Type IV pili are major bacterial virulence factors supporting adhesion, surface motility, and gene transfer. The polymeric pilus fiber is a highly dynamic molecular machine that switches between elongation and retraction. We used laser tweezers to investigate the dynamics of individual pili of Neisseria gonorrheae at clamped forces between 8 pN and 100 pN and at varying concentration of the retraction ATPase PilT. The elongation probability of individual pili increased with increasing mechanical force. Directional switching occurred on two distinct timescales, and regular stepping was absent on a scale > 3 nm. We found that the retraction velocity is bimodal and that the bimodality depends on force and on the concentration of PilT proteins. We conclude that the pilus motor is a multistate system with at least one polymerization mode and two depolymerization modes with the dynamics fine-tuned by force and PilT concentration. PMID- 19186151 TI - Distribution of frequencies of spontaneous oscillations in hair cells of the bullfrog sacculus. AB - Under in vitro conditions, free-standing hair bundles of the bullfrog (Rana catesbeiana) sacculus have exhibited spontaneous oscillations. We used a high speed complementary metal oxide semiconductor camera to track the active movements of multiple hair cells in a single field of view. Our techniques enabled us to probe for correlations between pairs of cells, and to acquire records on over 100 actively oscillating bundles per epithelium. We measured the statistical distribution of oscillation periods of cells from different areas within the sacculus, and on different epithelia. Spontaneous oscillations exhibited a peak period of 33 ms (+29 ms, -14 ms) and uniform spatial distribution across the sacculus. PMID- 19186153 TI - Kinetics of genetic switching into the state of bacterial competence. AB - Nonlinear amplification of gene expression of master regulators is essential for cellular differentiation. Here we investigated determinants that control the kinetics of the genetic switching process from the vegetative state (B-state) to the competent state (K-state) of Bacillus subtilis, explicitly including the switching window which controls the probability for competence initiation in a cell population. For individual cells, we found that after initiation of switching, the levels of the master regulator [ComK](t) increased with sigmoid shape and saturation occurred at two distinct levels of [ComK]. We analyzed the switching kinetics into the state with highest [ComK] and found saturation after a switching period of length 1.4 +/- 0.3 h. The duration of the switching period was robust against variations in the gene regulatory network of the master regulator, whereas the saturation levels showed large variations between individual isogenic cells. We developed a nonlinear dynamics model, taking into account low-number stochastic effects. The model quantitatively describes the probability and timescale of switching at the single cell level and explains why the ComK level in the K-state is highly sensitive to extrinsic parameter variations. Furthermore, the model predicts a transition from stochastic to deterministic switching at increased production rates of ComK in agreement with experimental data. PMID- 19186154 TI - Model of excitation-contraction coupling of rat neonatal ventricular myocytes. AB - The neonatal rat ventricular myocyte culture is one of the most popular experimental cardiac cell models. To our knowledge, the excitation-contraction coupling (ECC) of these cells, i.e., the process linking the electrical activity to the cytosolic Ca2+ transient and contraction, has not been previously analyzed, nor has it been presented as a complete system in detail. Neonatal cardiomyocytes are in the postnatal developmental stage, and therefore, the features of their ECC differ vastly from those of adult ventricular myocytes. We present the first complete analysis of ECC in these cells by characterizing experimentally the action potential and calcium signaling and developing the first mathematical model of ECC in neonatal cardiomyocytes that we know of. We show that in comparison to adult cardiomyocytes, neonatal cardiomyocytes have long action potentials, heterogeneous cytosolic Ca2+ signals, weaker sarcoplasmic reticulum Ca2+ handling, and stronger sarcolemmal Ca2+ handling, with a significant contribution by the Na+/Ca2+ exchanger. The developed model reproduces faithfully the ECC of rat neonatal cardiomyocytes with a novel description of spatial cytosolic [Ca2+] signals. Simulations also demonstrate how an increase in the cell size (hypertrophy) affects the ECC in neonatal cardiomyocytes. This model of ECC in developing cardiomyocytes provides a platform for developing future models of cardiomyocytes at different developmental stages. PMID- 19186155 TI - Mechanism of inhibition of the V-type molecular motor by tributyltin chloride. AB - Tributyltin chloride (TBT-Cl) is an endocrine disruptor found in many animal species, and it is also known to be an inhibitor for the V-ATPases that are emerging as potential targets in the treatment of diseases such as osteoporosis and cancer. We demonstrated by using biochemical and single-molecular imaging techniques that TBT-Cl arrests an elementary step for rotary catalysis of the V(1) motor domain. In the presence of TBT-Cl, the consecutive rotation of V(1) paused for a long duration ( approximately 0.5 s), even at saturated ATP concentrations, and the pausing positions were localized at 120 degrees intervals. Analysis of both the pausing time and moving time revealed that TBT-Cl has little effect on the binding affinity for ATP, but, rather, it arrests the catalytic event(s). This is the first report to demonstrate that an inhibitor arrests an elementary step for rotary catalysis of a V-type ATP-driven rotary motor. PMID- 19186156 TI - Versatile horizontal force probe for mechanical tests on pipette-held cells, particles, and membrane capsules. AB - We present a multipurpose nanomechanical force probe that combines a sideways mounted elastic cantilever and an optical-lever detection module with automated micropipette manipulation. It allows us to apply and measure compression, stretching, adhesion, and dissociation forces in the horizontal direction while providing a "side view" of ongoing experiments. The integrated micropipette setup facilitates the easy manipulation and mechanical interrogation of individual cells, functionalized particles, and synthetic membrane capsules. Pipette-held test objects are translated perpendicularly to and from the stationary cantilever, eliminating the need to attach them to a carrier surface and substantially reducing unwanted hydrodynamic coupling effects. Moreover, the test objects can be brought into contact with the cantilever anywhere along its length, which considerably enlarges the range of forces that can be applied with a single cantilever. Advantages of this instrument are demonstrated in example measurements of single-cell compression, membrane-tether extrusion, oligonucleotide stretching, and extraction of individual lipids from surfactant monolayer surfaces of microbubbles. PMID- 19186157 TI - Emotional pain without sensory pain--dream on? AB - The article by Danziger and colleagues in this issue of Neuron evaluates empathy in a unique population--individuals with congenital insensitivity to pain. As such, it provides insights into the brain's ability to evaluate others' feeling to observed pain without having a specific sensory experience of pain itself. PMID- 19186158 TI - Glia, adenosine, and sleep. AB - Sleep is regulated by a homeostatic process that has long been thought to involve adenosine (AD) originating from neurons. In this issue of Neuron, Halassa et al. present evidence that sleep homeostasis depends upon gliotransmission and associated accumulation of AD that dampens neuronal excitability. PMID- 19186159 TI - Silent synapses sit and wait for a better day. AB - Synaptic activity is thought to be critical for synaptic stabilization. In this issue of Neuron, Krishnaswamy and Cooper show that nicotinic synapses on autonomic neurons remain intact without synaptic activity. Postsynaptic responses are required, however, for presynaptic terminals to acquire the high-affinity choline transporter necessary for high-frequency transmission. PMID- 19186160 TI - MAP'ing CNS development and cognition: an ERKsome process. AB - The ERK MAP kinase signaling cascade plays critical roles in brain development, learning, memory, and cognition. It has recently been appreciated that mutation or deletion of elements within this signaling pathway leads to developmental syndromes in humans that are associated with impaired cognitive function and autism. Here, we review recent studies that provide insight into the biological roles of the ERKs in the brain that may underlie the cognitive deficits seen in these syndromes. PMID- 19186161 TI - The normalization model of attention. AB - Attention has been found to have a wide variety of effects on the responses of neurons in visual cortex. We describe a model of attention that exhibits each of these different forms of attentional modulation, depending on the stimulus conditions and the spread (or selectivity) of the attention field in the model. The model helps reconcile proposals that have been taken to represent alternative theories of attention. We argue that the variety and complexity of the results reported in the literature emerge from the variety of empirical protocols that were used, such that the results observed in any one experiment depended on the stimulus conditions and the subject's attentional strategy, a notion that we define precisely in terms of the attention field in the model, but that has not typically been completely under experimental control. PMID- 19186162 TI - Computational influence of adult neurogenesis on memory encoding. AB - Adult neurogenesis in the hippocampus leads to the incorporation of thousands of new granule cells into the dentate gyrus every month, but its function remains unclear. Here, we present computational evidence that indicates that adult neurogenesis may make three separate but related contributions to memory formation. First, immature neurons introduce a degree of similarity to memories learned at the same time, a process we refer to as pattern integration. Second, the extended maturation and change in excitability of these neurons make this added similarity a time-dependent effect, supporting the possibility that temporal information is included in new hippocampal memories. Finally, our model suggests that the experience-dependent addition of neurons results in a dentate gyrus network well suited for encoding new memories in familiar contexts while treating novel contexts differently. Taken together, these results indicate that new granule cells may affect hippocampal function in several unique and previously unpredicted ways. PMID- 19186163 TI - Can we share a pain we never felt? Neural correlates of empathy in patients with congenital insensitivity to pain. AB - Theories of empathy differ regarding the relative contributions of automatic resonance and perspective taking in understanding others' emotions. Patients with the rare syndrome of congenital insensitivity to pain cannot rely on "mirror matching" (i.e., resonance) mechanisms to understand the pain of others. Nevertheless, they showed normal fMRI responses to observed pain in anterior mid cingulate cortex and anterior insula, two key regions of the so-called "shared circuits" for self and other pain. In these patients (but not in healthy controls), empathy trait predicted ventromedial prefrontal responses to somatosensory representations of others' pain and posterior cingulate responses to emotional representations of others' pain. These findings underline the major role of midline structures in emotional perspective taking and understanding someone else's feeling despite the lack of any previous personal experience of it -an empathic challenge frequently raised during human social interactions. PMID- 19186164 TI - Astrocytic modulation of sleep homeostasis and cognitive consequences of sleep loss. AB - Astrocytes modulate neuronal activity by releasing chemical transmitters via a process termed gliotransmission. The role of this process in the control of behavior is unknown. Since one outcome of SNARE-dependent gliotransmission is the regulation of extracellular adenosine and because adenosine promotes sleep, we genetically inhibited the release of gliotransmitters and asked if astrocytes play an unsuspected role in sleep regulation. Inhibiting gliotransmission attenuated the accumulation of sleep pressure, assessed by measuring the slow wave activity of the EEG during NREM sleep, and prevented cognitive deficits associated with sleep loss. Since the sleep-suppressing effects of the A1 receptor antagonist CPT were prevented following inhibition of gliotransmission and because intracerebroventricular delivery of CPT to wild-type mice mimicked the transgenic phenotype, we conclude that astrocytes modulate the accumulation of sleep pressure and its cognitive consequences through a pathway involving A1 receptors. PMID- 19186165 TI - Mapping of class I and class II odorant receptors to glomerular domains by two distinct types of olfactory sensory neurons in the mouse. AB - The repertoire of approximately 1200 odorant receptors (ORs) is mapped onto the array of approximately 1800 glomeruli in the mouse olfactory bulb (OB). The spatial organization of this array is influenced by the ORs. Here we show that glomerular mapping to broad domains in the dorsal OB is determined by two types of olfactory sensory neurons (OSNs), which reside in the dorsal olfactory epithelium. The OSN types express either class I or class II OR genes. Axons from the two OSN types segregate already within the olfactory nerve and form distinct domains of glomeruli in the OB. These class-specific anatomical domains correlate with known functional odorant response domains. However, axonal segregation and domain formation are not determined by the class of the expressed OR protein. Thus, the two OSN types are determinants of axonal wiring, operate at a higher level than ORs, and contribute to the functional organization of the glomerular array. PMID- 19186166 TI - Pias3-dependent SUMOylation directs rod photoreceptor development. AB - Specification of retinal rod photoreceptors is determined by several different transcription factors that activate expression of rod-specific genes and repress expression of cone photoreceptor-specific genes. The mechanism by which this dual regulation occurs is unclear. We have found that Pias3, a transcriptional coregulator and E3 SUMO ligase that is selectively expressed in developing photoreceptors, promotes the differentiation of rod photoreceptors while preventing rods from adopting cone photoreceptor-like characteristics. Pias3 binds the photoreceptor-specific transcription factors Crx and Nr2e3 and is specifically targeted to the promoters of photoreceptor-specific genes. Pias3 SUMOylates Nr2e3, converting it into a potent repressor of cone-specific gene expression. Rod- and cone-specific promoters are bound by hyperSUMOylated proteins in rod photoreceptors, and blocking SUMOylation in photoreceptors results in cells with morphological and molecular features of cones and an absence of rod-specific markers. Our data thus identify Pias3-mediated SUMOylation of photoreceptor-specific transcription factors as a key mechanism of rod specification. PMID- 19186167 TI - Rapid functional maturation of nascent dendritic spines. AB - Spine growth and retraction with synapse formation and elimination plays an important role in shaping brain circuits during development and in the adult brain, yet the temporal relationship between spine morphogenesis and the formation of functional synapses remains poorly defined. We imaged hippocampal pyramidal neurons to identify spines of different ages. We then used two-photon glutamate uncaging, whole-cell recording, and Ca(2+) imaging to analyze the properties of nascent spines and their older neighbors. New spines expressed glutamate-sensitive currents that were indistinguishable from mature spines of comparable volumes. Some spines exhibited negligible AMPA receptor-mediated responses, but the occurrence of these "silent" spines was uncorrelated with spine age. In contrast, NMDA receptor-mediated Ca(2+) accumulations were significantly lower in new spines. New spines reconstructed using electron microscopy made synapses. Our data support a model in which outgrowth and enlargement of nascent spines is tightly coupled to formation and maturation of glutamatergic synapses. PMID- 19186169 TI - An activity-dependent retrograde signal induces the expression of the high affinity choline transporter in cholinergic neurons. AB - A well-accepted view of developing circuits is that synapses must be active to mature and persist, whereas inactive synapses remain immature and are eventually eliminated. We question this long-standing view by investigating nonfunctional cholinergic nicotinic synapses in the superior cervical ganglia (SCG) of mice with a disruption in the alpha3 nicotinic receptor (nAChR) subunit gene, a gene essential for fast synaptic transmission in sympathetic ganglia. Using imaging and electrophysiology, we show that synapses persist for at least 2-3 months without postsynaptic activity; however, the presynaptic terminals lack high affinity choline transporters (CHTs), and as a result, they are quickly depleted of transmitter. Moreover, we demonstrate with rescue experiments that CHT is induced by signals downstream of postsynaptic activity, converting immature terminals to mature terminals capable of sustaining transmitter release in response to high-frequency or continuous firing. Importantly, postsynaptic neurons must be continually active to maintain CHT in presynaptic terminals. PMID- 19186168 TI - Axon initial segment Ca2+ channels influence action potential generation and timing. AB - Although action potentials are typically generated in the axon initial segment (AIS), the timing and pattern of action potentials are thought to depend on inward current originating in somatodendritic compartments. Using two-photon imaging, we show that T- and R-type voltage-gated Ca(2+) channels are colocalized with Na(+) channels in the AIS of dorsal cochlear nucleus interneurons and that activation of these Ca(2+) channels is essential to the generation and timing of action potential bursts known as complex spikes. During complex spikes, where Na(+)-mediated spikelets fire atop slower depolarizing conductances, selective block of AIS Ca(2+) channels delays spike timing and raises spike threshold. Furthermore, AIS Ca(2+) channel block can decrease the number of spikelets within a complex spike and can even block single, simple spikes. Similar results were found in cortex and cerebellum. Thus, voltage-gated Ca(2+) channels at the site of spike initiation play a key role in generating and shaping spike bursts. PMID- 19186170 TI - Plasticity of burst firing induced by synergistic activation of metabotropic glutamate and acetylcholine receptors. AB - Subiculum, the primary efferent pathway of hippocampus, participates in memory for spatial tasks, relapse to drug abuse, and temporal lobe seizures. Subicular pyramidal neurons exhibit low-threshold burst firing driven by a spike afterdepolarization. Here we report that burst firing can be regulated by stimulation of afferent projections to subiculum. Unlike synaptic plasticity, burst plasticity did not require synaptic depolarization, activation of AMPA or NMDA receptors, or action potential firing. Rather, enhancement of burst firing required synergistic activation of group I, subtype 1 metabotropic glutamate receptors (mGluRs) and muscarinic acetylcholine receptors (mAChR). When either of these receptors was blocked, a suppression of bursting was revealed, which in turn was blocked by antagonists of group I, subtype 5 mGluRs. These results indicate that the output of subiculum can be strongly and bidirectionally regulated by activation of glutamatergic inputs within the hippocampus and cholinergic afferents from the medial septum. PMID- 19186171 TI - The excitatory neuronal network of the C2 barrel column in mouse primary somatosensory cortex. AB - Local microcircuits within neocortical columns form key determinants of sensory processing. Here, we investigate the excitatory synaptic neuronal network of an anatomically defined cortical column, the C2 barrel column of mouse primary somatosensory cortex. This cortical column is known to process tactile information related to the C2 whisker. Through multiple simultaneous whole-cell recordings, we quantify connectivity maps between individual excitatory neurons located across all cortical layers of the C2 barrel column. Synaptic connectivity depended strongly upon somatic laminar location of both presynaptic and postsynaptic neurons, providing definitive evidence for layer-specific signaling pathways. The strongest excitatory influence upon the cortical column was provided by presynaptic layer 4 neurons. In all layers we found rare large amplitude synaptic connections, which are likely to contribute strongly to reliable information processing. Our data set provides the first functional description of the excitatory synaptic wiring diagram of a physiologically relevant and anatomically well-defined cortical column at single-cell resolution. PMID- 19186172 TI - Temporal coherence in the perceptual organization and cortical representation of auditory scenes. AB - Just as the visual system parses complex scenes into identifiable objects, the auditory system must organize sound elements scattered in frequency and time into coherent "streams." Current neurocomputational theories of auditory streaming rely on tonotopic organization of the auditory system to explain the observation that sequential spectrally distant sound elements tend to form separate perceptual streams. Here, we show that spectral components that are well separated in frequency are no longer heard as separate streams if presented synchronously rather than consecutively. In contrast, responses from neurons in primary auditory cortex of ferrets show that both synchronous and asynchronous tone sequences produce comparably segregated responses along the tonotopic axis. The results argue against tonotopic separation per se as a neural correlate of stream segregation. Instead we propose a computational model of stream segregation that can account for the data by using temporal coherence as the primary criterion for predicting stream formation. PMID- 19186173 TI - Reversible coupling of individual phycobiliprotein isoforms during state transitions in the cyanobacterium Trichodesmium analysed by single-cell fluorescence kinetic measurements. AB - In the non-heterocyst, marine cyanobacterium Trichodesmium nitrogen fixation is confined to the photoperiod and occurs coevally with oxygenic photosynthesis although nitrogenase is irreversibly inactivated by oxygen. In previous studies it was found that regulation of photosynthesis for nitrogen fixation involves Mehler reaction and various activity states with reversible coupling of photosynthetic components. We now investigated these activity states in more detail. Spectrally resolved fluorescence kinetic measurements of single cells revealed that they were related to alternate uncoupling and coupling of phycobilisomes from and to the photosystems, changing the effective cross-section of PSII. Therefore, we isolated and purified the phycobiliproteins of Trichodesmium via ion exchange chromatography and recorded their UV/VIS absorption, fluorescence excitation and fluorescence emission spectra. After describing these spectra by mathematical equations via the Gauss-Peak-Spectra method, we used them to deconvolute the in vivo fluorescence spectra of Trichodesmium cells. This revealed that the contribution of different parts of the phycobilisome antenna to fluorescence quenching changed during the daily activity cycle, and that individual phycobiliproteins can be reversibly coupled to the photosystems, while the expression levels of these proteins did not change much during the daily activity cycle. Thus we propose that variable phycobilisome coupling plays a key role in the regulation of photosynthesis for nitrogen fixation in Trichodesmium. PMID- 19186174 TI - The experimental chemotherapeutic N6-furfuryladenosine (kinetin-riboside) induces rapid ATP depletion, genotoxic stress, and CDKN1A(p21) upregulation in human cancer cell lines. AB - Cytokinins and cytokinin nucleosides are purine derivatives with potential anticancer activity. N(6)-furfuryladenosine (FAdo, kinetin-riboside) displays anti-proliferative and apoptogenic activity against various human cancer cell lines, and FAdo has recently been shown to suppress tumor growth in murine xenograft models of human leukemia and melanoma. In this study, FAdo-induced genotoxicity, stress response gene expression, and cellular ATP depletion were examined as early molecular consequences of FAdo exposure in MiaPaCa-2 pancreas carcinoma, A375 melanoma, and other human cancer cell lines. FAdo, but not adenosine or N(6)-furfuryladenine (FA), displayed potent anti-proliferative activity that was also observed in human primary fibroblasts and keratinocytes. Remarkably, massive ATP depletion and induction of genotoxic stress as assessed by the alkaline comet assay occurred within 60-180min of exposure to low micromolar concentrations of FAdo. This was followed by rapid upregulation of CDKN1A and other DNA damage/stress response genes (HMOX1, DDIT3, and GADD45A) as revealed by expression array and Western analysis. Pharmacological and siRNA based genetic inhibition of adenosine kinase (ADK) suppressed FAdo cytotoxicity and also prevented ATP depletion and p21 upregulation suggesting the importance of bioconversion of FAdo into the nucleotide form required for drug action. Taken together our data suggest that early induction of genotoxicity and energy crisis are important causative factors involved in FAdo cytotoxicity. PMID- 19186175 TI - Evaluation of urinary nucleosides in breast cancer patients before and after tumor removal. AB - OBJECTIVE: Evaluation of altered urinary nucleosides before and after tumor removal of breast cancer (BCa). DESIGN AND METHODS: Targeted metabolite profiling of 14 urinary nucleosides was conducted with both pre- and post-operative female patients with BCa (n=150, age: 46.6+/-7.7 years), and female controls (n=150, age: 46.8+/-7.7 years) by liquid chromatography-tandem mass spectrometry coupled to on-line extraction. RESULTS: Levels of modified nucleosides (5-hydroxymethyl 2'-deoxyuridine, P<0.001; 8-hydroxy-2'-deoxyguanosine, P<0.001; 1 methyladenosine, P<0.02; N(2),N(2)-dimethylguanosine, P<0.001) were significantly higher in pre-operative patients than in both normal controls and post-operative patients. CONCLUSIONS: This approach could be used to further understand the pathogenesis of BCa as well as to evaluate the effects of medical treatment. PMID- 19186176 TI - Nanoparticle-based targeted drug delivery. AB - Nanotechnology could be defined as the technology that has allowed for the control, manipulation, study, and manufacture of structures and devices in the "nanometer" size range. These nano-sized objects, e.g., "nanoparticles", take on novel properties and functions that differ markedly from those seen from items made of identical materials. The small size, customized surface, improved solubility, and multi-functionality of nanoparticles will continue to open many doors and create new biomedical applications. Indeed, the novel properties of nanoparticles offer the ability to interact with complex cellular functions in new ways. This rapidly growing field requires cross-disciplinary research and provides opportunities to design and develop multifunctional devices that can target, diagnose, and treat devastating diseases such as cancer. This article presents an overview of nanotechnology for the biologist and discusses the attributes of our novel XPclad((c)) nanoparticle formulation that has shown efficacy in treating solid tumors, single dose vaccination, and oral delivery of therapeutic proteins. PMID- 19186179 TI - Rescue of the neuroblastoma mutant of the human nucleoside diphosphate kinase A/nm23-H1 by the natural osmolyte trimethylamine-N-oxide. AB - The point mutation S120G in human nucleoside diphosphate kinase A, identified in patients with neuroblastoma, causes a protein folding defect. The urea-unfolded protein cannot refold in vitro, and accumulates as a molten globule folding intermediate. We show here that the trimethylamine-N-oxide (TMAO) corrects the folding defect and stimulated subunit association. TMAO also substantially increased the stability to denaturation by urea of both wild-type and S120G mutant. A non-native folding intermediate accumulated in the presence of 4.5-7M urea and of 2M TMAO. It was inactive, monomeric, contained some secondary structure but no tertiary structure and displayed a remarkable stability to denaturation. PMID- 19186178 TI - Activation of PI3K/Akt signaling has a dominant negative effect on IL-12 production by macrophages infected with Leishmania amazonensis promastigotes. AB - Infection of macrophages with Leishmania parasites does not result in the production of IL-12. In addition, infection with Leishmania suppresses IL-12 production elicited by otherwise potent activators of IL-12. We provide evidence that engagement of phosphatidyl inositol-3 kinase (PI3K) signaling during Leishmania amazonensis infection leads to the prevention of IL-12 p70 production at the level of transcription of its p40 subunit in bone marrow derived macrophages (BMDMPhi). Inhibition of PI3K signaling with specific inhibitors of PI3K or the downstream kinase Akt, reverses the IL-12 blockade. Although the MAP kinase ERK (p44 and p42) was transiently activated by infection with L. amazonensis, inhibition of MEK, the kinase upstream of ERK, with PD98059, did not reverse the blockade of IL-12. Furthermore, inhibition of the other MAP kinases JNK and p38 as well as treatment of cells with pertussis toxin that blocks G protein mediated signaling, did not reverse the prevention of IL-12 production by Leishmania infection. Interestingly, activation of PI3K/Akt signaling had differential effects on ERK and p38 activation. Taken together we propose that infection of BMDMPhi with Leishmania promastigotes activates both positive and negative signaling pathways that control IL-12 production. PI3K signaling activated by the infection is the negative signaling pathway that prevents IL-12 production. PMID- 19186177 TI - Mechanisms of macrophage migration inhibitory factor (MIF)-dependent tumor microenvironmental adaptation. AB - Since its activity was first reported in the mid-1960s, macrophage migration inhibitory factor (MIF) has gone from a cytokine activity modulating monocyte motility to a pleiotropic regulator of a vast array of cellular and biological processes. Studies in recent years suggest that MIF contributes to malignant disease progression on several different levels. Both circulating and intracellular MIF protein levels are elevated in cancer patients and MIF expression reportedly correlates with stage, metastatic spread and disease-free survival. Additionally, MIF expression positively correlates with angiogenic growth factor expression, microvessel density and tumor-associated neovascularization. Not coincidentally, MIF has recently been shown to contribute to tumoral hypoxic adaptation by promoting hypoxia-induced HIF-1alpha stabilization. Intriguingly, hypoxia is a strong regulator of MIF expression and secretion, suggesting that hypoxia-induced MIF acts as an amplifying factor for both hypoxia and normoxia-associated angiogenic growth factor expression in human malignancies. Combined, these findings suggest that MIF overexpression contributes to tumoral hypoxic adaptation and, by extension, therapeutic responsiveness and disease prognosis. This review summarizes recent literature on the contributions of MIF to tumor-associated angiogenic growth factor expression, neovascularization and hypoxic adaptation. We also will review recent efforts aimed at identifying and employing small-molecule antagonists of MIF as a novel approach to cancer therapeutics. PMID- 19186180 TI - Genome-wide validation of Magnaporthe grisea gene structures based on transcription evidence. AB - Accurate cDNA data is useful to validate gene structures in a genome. We sequenced 35189 expressed sequence tags (ESTs) obtained from the highly destructive rice blast fungus, Magnaporthe grisea. Our custom-made computational programs mapped these ESTs on the M. grisea genome sequence, and reconstructed gene structures as well as protein-coding regions. As a result, we predicted 4480 protein-coding sequences, which were more accurate than ab initio predictions. Moreover, cross-species comparisons suggested that our predicted proteins were nearly complete. The cDNA clones obtained in this study will be important for further experimental studies. Our genome annotation is available at http://www.mg.dna.affrc.go.jp/. PMID- 19186182 TI - Targeting gene delivery to activated vascular endothelium using anti E/P-Selectin antibody linked to PAMAM dendrimers. AB - INTRODUCTION: The adhesion molecules P- and E-selectin are expressed on activated endothelial cells, and are good targets for gene therapy aimed at inflammatory disease. We have therefore investigated the potential of targeting PAMAM dendrimers, a non viral vector system, to cells expressing P/E-selectin using a monoclonal antibody that recognises these molecules. MATERIALS AND METHODS: We used biotin and avidin to cross-link anti E/P-Selectin monoclonal antibody to pre formed Superfect-DNA complexes that were then used to transfect reporter genes to CHO cells expressing E-Selectin, cytokine-activated primary Human Saphenous Vein Endothelial Cells (HSVEC) and whole vein segments. RESULTS: The use of the anti E/P-Selectin antibody increased the transfection efficiency in CHO-E cells, activated HSVEC and saphenous vein segments ex vivo. We also showed that the antibody improved the binding of the complexes onto cells as well as the internalisation kinetics. DISCUSSION: We demonstrate here that by attaching antibodies onto PAMAM dendrimers the efficiency of transfection can be significantly improved in cells or tissues expressing the receptor. This technology has potential in the treatment of cardiovascular disease by gene therapy but can also be used with different antibodies to target other diseased cells or tissues. PMID- 19186181 TI - Colorectal cancer expression of peroxisome proliferator-activated receptor gamma (PPARG, PPARgamma) is associated with good prognosis. AB - BACKGROUND & AIMS: The peroxisome proliferator-activated receptor gamma (PPARG, PPARgamma) is a nuclear receptor that regulates expression of mediators of lipid metabolism and the inflammatory response. There is controversy over the pro oncogenic or antioncogenic effects of PPARG, and little is known about its prognostic significance in colon cancer. METHODS: Among 470 patients with colorectal cancer (stages I-IV) identified in 2 independent prospective cohorts, PPARG expression was detected in 102 tumors (22%) by immunohistochemistry. Cox proportional hazards models were used to compute hazard ratios (HRs) of colorectal cancer-specific and overall mortalities, adjusted for patient characteristics and molecular features including cyclooxygenase 2, fatty acid synthase, KRAS, BRAF, PIK3CA, p53, p21, beta-catenin, LINE-1 hypomethylation, microsatellite instability (MSI), and the CpG island methylation phenotype (CIMP). RESULTS: Compared with patients with PPARG-negative tumors, patients with PPARG-positive tumors had significantly lower overall mortality, determined by Kaplan-Meier analysis (P=.0047), univariate Cox regression (HR, 0.55; 95% confidence interval [CI], 0.37-0.84; P=.0053), and multivariate analysis (adjusted HR, 0.43; 95% CI, 0.27-0.69; P=.0004). Patients with PPARG-positive tumors experienced lower colorectal cancer-specific mortality (adjusted HR, 0.44; 95% CI, 0.25-0.79; P=.0054). The relationship between PPARG and lower mortality did not appear to be significantly modified by MSI, CIMP, LINE-1, or the other clinical and molecular variables examined (all P(interaction)>.05). CONCLUSIONS: Tumor expression of PPARG is independently associated with longer survival of patients. PPARG expression appears to mark an indolent subset of colorectal cancers. PMID- 19186183 TI - A novel plasmid DNA electroporation method allows transfection of murine DC. AB - Under steady state conditions dendritic cells (DC) exert tolerogenic function, but acquire potent immunogenic function due to strong upregulation of costimulatory molecules and proinflammatory cytokines. In numerous studies the potential of modified DC to induce tolerance or immune reactions towards a distinct antigen has been demonstrated. However, DC are refractory to transfection with plasmid DNA by non-viral methods. In this study we have tested the suitability of a newly developed electroporation device to transfect immature murine bone-marrow derived DC (BM-DC). Transfected BM-DC expressed reporter molecules at considerable extent which renders this method suitable to perform all kinds of promoter studies. While electroporation did not alter the low allostimulatory capacity of immature BM-DC, it impaired the stimulation associated increase in allostimulatory potency of transfectants. However, stimulated transfected BM-DC pulsed with myelin oligodendrocyte protein (MOG) derived peptide induced proliferation of MOG-reactive CD4(+) T cells as potently as did non-transfected MOG peptide-pulsed BM-DC. BM-DC transfected with an expression construct encoding MOG efficiently stimulated MOG peptide-specific T cell proliferation. Transfection of BM-DC with an IL-10 encoding expression construct resulted in high IL-10 expression and strongly diminished allogeneic T cell proliferation. Therefore, this method also allows to study functional properties of genetically altered DC. PMID- 19186184 TI - Clock genes period and timeless are rhythmically expressed in brains of newly hatched, photosensitive larvae of the fly, Sarcophaga crassipalpis. AB - While roles of the clock genes period (per) and timeless (tim) are relatively well understood in relation to circadian clocks, their potential roles in insect photoperiodism remain enigmatic. In this study, the expression of per and tim genes under two contrasting photoperiods is described in the central nervous system of photoperiodically sensitive, newly hatched first instar larvae of the flesh fly, Sarcophaga crassipalpis. Using qPCR, diel oscillations were observed in the mRNA levels of both genes under long-day (15 h light:9h dark, promotes direct development) and short-day conditions (11 h light:13 h dark, induces pupal diapause). Peak per and tim mRNA oscillations were closely associated with the light/dark transition. The conspicuous difference between the two photoperiodic conditions was that the sharp increase in per and tim mRNA abundance occurred during the light phase under long days but during the dark phase under short days. The diel oscillations were, at least in part, driven by an endogenous component, as demonstrated by transferring larvae to continuous darkness. The cells displaying Tim- and Per-like immunoreactivities (Tim- and Per-LIRs) were localized using anti-Drosophila-Per and anti-Chymomyza-Tim antibodies. Per-LIR and Tim-LIR co-localized in three groups of cells in each brain hemisphere. Two other groups, one in the brain hemispheres and the other in the fused ventral nerve ganglion, expressed only the Per-LIR. PMID- 19186185 TI - Helicoverpa armigera (Lepidoptera: Noctuidae) larvae that survive sublethal doses of nucleopolyhedrovirus exhibit high metabolic rates. AB - To determine the effect of sublethal doses of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HearSNPV) on the metabolic rate of H. armigera, the respiration rates of third instar H. armigera larvae inoculated with sublethal doses of HearSNPV were evaluated. Respiration rates, measured as the rate of CO(2) production (VCO(2)), were recorded daily using closed-system respirometry. By 4 days post-inoculation (dpi), the metabolic rates of LD(25) or LD(75) survivors were significantly higher than that of uninoculated controls. When dose data were pooled, the VCO(2) values of larvae that survived inoculation (0.0288mlh(-1)), the uninoculated controls (0.0250mlh(-1)), and the larvae that did not survive inoculation (0.0199mlh(-1)) differed significantly from one another. At 4dpi, the VCO(2) of the uninoculated controls were significantly lower than the VCO(2) of inoculation survivors, but significantly higher than the VCO(2) of inoculation non-survivors. Inoculation survivors may have had high metabolic rates due to a combination of viral replication, organ damage, and an energy-intensive induced cellular immune response. The high 4dpi metabolic rate of inoculation survivors may reflect an effective immune response and may be seen as the metabolic signature of larvae that are in the process of surviving inoculation with HearSNPV. PMID- 19186186 TI - Nutrients in fruit increase fertility in wild-caught females of large and long lived Euphaedra species (Lepidoptera, Nymphalidae). AB - Fruit-feeding butterflies can experience a more nutrient rich adult diet than nectar-feeding species, and can be expected to use these nutrients for egg production. Here we compare life span, and reproduction parameters of wild-caught females of large and long-lived species on either a sucrose or a mashed banana diet. With small sample sizes per species, but rich longitudinal data for each individual, we examined the longitudinal reproduction pattern, egg size and hatchability of these butterflies in captivity. Diet significantly affected mortality in captivity in a time-dependent manner. On average, we found that butterflies fed mashed banana laid 1.855 times more eggs than those fed sugar. They laid significantly more eggs when they laid and conserved egg size with age while butterflies fed sucrose showed significantly declining egg sizes. Egg hatchability was not significantly affected by diet. Long pre-oviposition periods, significantly smaller first eggs, and absence of age at capture effects on intensity of reproduction indicate low reproduction rates in the field that are due to low food availability. With our small sample sizes, we did not detect significant differences between the species in their response to the diet treatments. PMID- 19186187 TI - Length variation in a specific region of the period gene correlates with differences in pupal diapause incidence in the flesh fly, Sarcophaga bullata. AB - We report differences in the length of a specific region of the circadian clock gene period (per) that correlate with different capacities for pupal diapause in the flesh fly, Sarcophaga bullata. The conspicuous difference is located in a region we refer to as the putative C-terminal photoperiodic (CP) region. The length of the CP region correlates inversely with the incidence of diapause. A deletion of 33 amino acids in this region correlates with a significant increase in the incidence of diapause, from 78.1% to 93.0%, and an insertion of 9 amino acids in the same area correlates with a drop in the diapause incidence to 4.0%. This correlation suggests a possible functional role for this region of per in photoperiodism. PMID- 19186188 TI - Mathematical simulations of ligand-gated and cell-type specific effects on the action potential of human atrium. AB - In the mammalian heart, myocytes and fibroblasts can communicate via gap junction, or connexin-mediated current flow. Some of the effects of this electrotonic coupling on the action potential waveform of the human ventricular myocyte have been analyzed in detail. The present study employs a recently developed mathematical model of the human atrial myocyte to investigate the consequences of this heterogeneous cell-cell interaction on the action potential of the human atrium. Two independent physiological processes which alter the physiology of the human atrium have been studied. i) The effects of the autonomic transmitter acetylcholine on the atrial action potential have been investigated by inclusion of a time-independent, acetylcholine-activated K(+) current in this mathematical model of the atrial myocyte. ii) A non-selective cation current which is activated by natriuretic peptides has been incorporated into a previously published mathematical model of the cardiac fibroblast. These results identify subtle effects of acetylcholine, which arise from the nonlinear interactions between ionic currents in the human atrial myocyte. They also illustrate marked alterations in the action potential waveform arising from fibroblast-myocyte source-sink principles when the natriuretic peptide-mediated cation conductance is activated. Additional calculations also illustrate the effects of simultaneous activation of both of these cell-type specific conductances within the atrial myocardium. This study provides a basis for beginning to assess the utility of mathematical modeling in understanding detailed cell-cell interactions within the complex paracrine environment of the human atrial myocardium. PMID- 19186189 TI - The drug monosodium luminol (GVT) preserves crypt-villus epithelial organization and allows survival of intestinal T cells in mice infected with the ts1 retrovirus. AB - Of the cytopathic retroviruses that affect mammals, including HIV-1, many selectively infect CD4+ T cells and cause immunosuppressive syndromes. These diseases destroy both the thymus and the small and large intestines, after infecting and killing T-lineage cells in both tissues. A mutant of the murine leukemia retrovirus MoMuLV-TB, called ts1, causes this syndrome in susceptible strains of mice. In FVB/N strain mice that are infected at birth, thymic atrophy, CD4+ T cell loss, intestinal collapse, body wasting, and death occur by approximately 30-40 days postinfection (dpi). Apoptosis of ts1-infected T-lineage cells, in the thymus, peripheral lymphoid system and intestines is caused by accumulation of the ts1 mutant viral envelope preprotein gPr80(env), which is inefficiently cleaved into the mature viral proteins gp70 and PrP15E. We show here that ts1 infection in the small intestine is followed by loss of intestinal epithelial cell (IEC) thyroid-stimulating hormone (TSH) and cell cycling gradients (along the crypt-villus axes), accumulation of gPr80(env) in intestinal cells, apoptosis of developing T cells in the lamina propria (LP), and intestinal collapse by approximately 30 dpi. In infected mice treated with the antioxidant drug monosodium luminol (GVT), however, normal intestinal epithelial cell gradients are still in place at 30 dpi, and IECs covering both the crypts and villi contain large amounts of the antioxidant transcription factor Nrf2. In addition, no apoptotic cells are present, and accumulated gpr80(env) is absent from the tissue at this time. We conclude that GVT treatment can make ts1 a noncytopathic virus for intestinal lymphoid cells, as it does for thymocytes [25]. As in the thymus, GVT may protect the intestine by reducing oxidant stress in infected intestinal T cells, perhaps by prevention of gPr80(env) accumulation via Nrf2 upregulation in the IECs. These results identify GVT as a potential therapy for intestinal diseases or inflammatory conditions, including HIV-AIDS, in which oxidative stress is a triggering or exacerbating factor. PMID- 19186190 TI - Comparative study of the replication of infectious bursal disease virus in DF-1 cell line and chicken embryo fibroblasts evaluated by a new real-time RT-PCR. AB - A real-time RT-PCR method was developed for the detection of infectious bursal disease virus (IBDV). The VP5 gene of IBDV was chosen as the target binding region for a specific TaqMan probe. The results showed that viral genomic copy number could be quantified accurately ranging from 10(8)copies/microL to 10(1)copies/microL. No positive signal was detected for other avian pathogens in the specificity test. This assay was highly sensitive and could detect as little as 30 copies of viral RNA. Both the coefficients of variation (CVs) of inter- and intra-assay reproducibility were less than 2%. Growth curves of the IBDV Gt strain in chicken embryo fibroblasts (CEF) and DF-1 cells were evaluated by the real-time RT-PCR. The data showed that the cytopathic effects of the virus in CEF and DF-1 cells were similar. However, higher viral titers were detected in the DF 1 cell line. This study indicated that the real-time RT-PCR approach provided a powerful diagnostic tool with high sensitivity and specificity for the identification and quantitation of IBDV. The DF-1 cell line may be a more suitable continuous cell line for the propagation of IBDV compared to CEF. PMID- 19186191 TI - Evaluation of different RT enzyme standards for quantitation of retroviruses using the single-tube fluorescent product-enhanced reverse transcriptase assay. AB - PCR-based reverse transcriptase (RT) assays are highly sensitive for broad detection of retroviruses. These assays are currently used for demonstrating the absence of retroviral contamination in vaccines and can also be applied to clinical and laboratory research to investigate low-virus replication. A single tube fluorescent product-enhanced reverse transcriptase assay (STF-PERT) has been published that was highly sensitive for retrovirus detection (<10 virions), with enhanced reproducibility and increased efficiency [Sears, J.F., Khan, A.S., 2003. Single-tube fluorescent product-enhanced reverse transcriptase assay with AmpliWax (STF-PERT) for retrovirus quantitation. J. Virol. Meth. 108, 139-142]. In this report, the step-by-step setup and performance of the STF-PERT assay is described and sensitivity, reproducibility and specificity of the assay reported using three different RTs as standards: avian myeloblastosis virus (AMV) RT, murine leukemia virus (MMLV) RT, and human immunodeficiency virus type 1 (HIV-1) RT. Evaluation of virus stocks showed about 1-2 logs difference in RT detection and retrovirus quantitation with the different RT enzyme standards; in general, virus determination using HIV-1 RT was comparable to using the relevant virus RT. PMID- 19186192 TI - Development of a loop-mediated isothermal amplification for rapid detection of orf virus. AB - A loop-mediated isothermal amplification (LAMP) assay using six primers targeting a highly conserved region of the B2L gene has been developed to diagnose orf virus. The assay produces a ladder-like pattern of products on an agarose gel that can be specifically digested with BsrGI enzyme. The sensitivity of the LAMP assay, which was determined to be a single copy of the standard plasmid, was 100 fold and 10 fold higher than PCR and nested PCR, respectively; furthermore, no cross-reactivity was founded with the other tested viruses. By staining the products directly in the tube with PicoGreen or ethidium bromide, the products can be visualized with a similar sensitivity as by gel electrophoresis. Clinical samples were tested using PCR, nested PCR and LAMP assay and the positive rates were 60%, 70% and 70%, respectively. The LAMP assay allows easy, rapid, accurate and sensitive detection of infection with orf virus and is especially applicable in a resource-limited situation. PMID- 19186193 TI - Rapid detection of human immunodeficiency virus type 1 group M by a reverse transcription-loop-mediated isothermal amplification assay. AB - A rapid one-step reverse transcription-loop-mediated isothermal amplification (RT LAMP) assay targeting the pol-integrase gene was developed to detect human immunodeficiency virus type 1 (HIV-1) group M. This HIV-1 RT-LAMP assay is simple and rapid, and amplification can be completed within 35min under isothermal conditions at 60 degrees C. The 100% detection limit of HIV-1 RT-LAMP was determined using a standard strain (WHO HIV-1 [97/656]) in octuplicate and found to be 120 copies/ml. The RT-LAMP assay was evaluated for use for clinical diagnosis using plasma samples collected from 57 HIV-1-infected and 40 uninfected individuals in Cameroon, where highly divergent HIV-1 strains are prevalent. Of the 57 samples from infected individuals, 56 harbored group-M HIV-1 strains, such as subtypes A, B, G, F2, and circulating recombinant forms (CRFs) _01, _02, _09, _11, _13; all were RT-LAMP positive. One sample harboring group-O HIV-1 and the 40 HIV-1-uninfected samples were RT-LAMP negative. These findings indicate that HIV-1 RT-LAMP can detect HIV-1 group-M RNA from plasma samples rapidly and with high sensitivity and specificity. These data also suggest that this RT-LAMP assay can be useful for confirming HIV diagnosis, particularly in resource-limited settings. PMID- 19186194 TI - CpG oligonucleotides for immunotherapeutic treatment of neuroblastoma. AB - Neuroblastoma is the most common extracranial solid tumor malignancy of childhood. Although it is generally responsive to treatment, high risk cases of neuroblastoma frequently recur. The prognosis for relapsed cases is extremely poor despite aggressive therapy. The frequency of relapse and subsequent failure of further treatment has spurred the need to develop non toxic and more effective treatments for targeting residual tumor cells during the phase of minimal residual disease. Traditional cancer therapies are non-specific, leading to the destruction of normal, healthy tissues. Failure to induce specific tumor immunity may be due to several immunosuppressive factors. Primary amongst these factors are: lack of co-stimulatory molecules on the surface of tumor cells, the ability of the tumor to modulate immunity in a suppressive manner and the presence of an immunosuppressive microenvironment at the location of the tumor. Unfortunately, tumor tolerance impedes the ability to establish immunity to tumor antigens and overcoming this tolerance is essential to developing effective tumor immunity. Vaccine strategies that target host immune effector cells with synthetic oligodeoxynucleotides (ODNs) that contain unmethylated CpG motifs (CpG-ODNs) represent a novel approach to overcoming tolerance in cancer therapy. This approach enables biasing of host immunity toward a proinflammatory Th1 and thus anti-tumor response. The addition of immunogenic tumor specific antigen to the CpG-ODN vaccine may allow for specific targeting and killing of established tumors. PMID- 19186195 TI - Response to "Analysis of adverse events of potential autoimmune aetiology in a large integrated safety database of AS04 adjuvanted vaccines" by Verstraeten et al. PMID- 19186196 TI - Rough mutants defective in core and O-polysaccharide synthesis and export induce antibodies reacting in an indirect ELISA with smooth lipopolysaccharide and are less effective than Rev 1 vaccine against Brucella melitensis infection of sheep. AB - Classical brucellosis vaccines induce antibodies to the O-polysaccharide section of the lipopolysaccharide that interfere in serodiagnosis. Brucella rough (R) mutants lack the O-polysaccharide but their usefulness as vaccines is controversial. Here, Brucella melitensis R mutants in all main lipopolysaccharide biosynthetic pathways were evaluated in sheep in comparison with the reference B. melitensis Rev 1 vaccine. In a first experiment, these mutants were tested for ability to induce anti-O-polysaccharide antibodies, persistence and spread through target organs, and innocuousness. Using the data obtained and those of genetic studies, three candidates were selected and tested for efficacy as vaccines against a challenge infecting 100% of unvaccinated ewes. Protection by R vaccines was 54% or less whereas Rev 1 afforded 100% protection. One-third of R mutant vaccinated ewes became positive in an enzyme-linked immunosorbent assay with smooth lipopolysaccharide due to the core epitopes remaining in the mutated lipopolysaccharide. We conclude that R vaccines interfere in lipopolysaccharide immunosorbent assays and are less effective than Rev 1 against B. melitensis infection of sheep. PMID- 19186197 TI - Susceptibility of the Aotus nancymaae owl monkey to eastern equine encephalitis. AB - Eastern equine encephalitis virus (EEEV) is an arthropod-borne virus associated with life-threatening encephalitis in humans, equines, birds and many other domestic animals. To investigate the suitability of the Aotus nancymaae New World owl monkey as a viable animal model for EEE candidate vaccine testing we used clinical presentation, serology, viral isolation and PCR to evaluate pathogenesis and immunity in infected animals. Monkeys were inoculated subcutaneously (SQ) or intranasally (IN) with 10(4)pfu of virulent EEEV and were initially followed for 45 days. While none of the animals displayed clinical signs of disease, all of the SC inoculated animals (n=6) manifested a viremia averaging 3.2 days (+/-0.8 days). Likewise, serologic responses (IgM, IgG and PRNT) were observed in all SC infected animals. Interestingly, none of the IN inoculated animals (n=6) became viremic or mounted an antibody response and no pathological abnormalities were observed in two animals that were necropsied on day 6 post-infection (p.i.) from each group. To determine if the antibodies produced by the SC inoculated animals were protective against homologous challenge, three animals from the SC group were serologically evaluated on day 253 p.i. and were administered an inoculum identical to initial challenge on day 270 p.i. A positive control group of four naive animals was also infected as before. All of the naive positive control animals manifested a similar viremia as observed initially, averaging 2.75 days (+/-0.5 days) while none of the previously challenged animals became viremic. On days 45 and 253 p.i. geometric mean PRNT titers in the SC group were 453 and 101, respectively. This study demonstrates that the Aotus nancymaae can be reproducibly infected with EEE virus and can serve as a suitable model for infection and immunogenicity for the evaluation of candidate vaccines against EEEV. PMID- 19186198 TI - Importance of subunit vaccine antigen of major Fli C antigenic site of Salmonella enteritidis II: a challenge trial. AB - Salmonella enterica subsp. enterica serovar Enteritidis (SE) infection in chickens shows a mild pathogenicity except for young ages, compared with other animals, and laying hens sometimes produce SE-contaminated eggs leading to public health concerns. To reduce the problem, SE bacterin in poultry farms has been applied. We previously demonstrated that a subunit antigen, g.m. part polypeptide in SE-Fli C (SEp 9), could be a candidate subunit antigen of SE vaccine which may show less side effects in chickens. In this study, we used SEp 9 along with an adjuvant to inoculate chickens, then the chickens were orally challenged with SE, and suppression of the SE count in the cecum was investigated. Chickens inoculated with a commercial SE vaccine were prepared as positive controls (vaccine group), and those with physiological saline (control group) for comparison of the bacterial count after challenge. Employing two types of antibody-detection ELISA coated with either de-flagellated SE or SEp 9, specific antibody levels in blood and the intestine were determined. The bacterial count was significantly lower 1 and 3 weeks after challenge in the SEp 9 than in the control group. Specific antibody only against SEp 9 in blood but not the intestine of these birds in the SEp 9 group was detected. This study confirmed that SEp 9 antigen is a major effective antigen in SE inactivated vaccine, and it is suggested that only the subunit vaccine antigen SEp 9 is needed to effectively suppress colonization in the chicken intestine, without the need for other SE component antigens. PMID- 19186199 TI - Exosomes are an effective vaccine against congenital toxoplasmosis in mice. AB - Toxoplasmosis is a serious disease in humans and may cause abortion or congenital infection if a woman is exposed to the disease for the first time during pregnancy. Infection before pregnancy normally results in immunity protecting the foetus, suggesting that it may be possible to block vertical transmission of the parasite by appropriate vaccination before pregnancy. We found that the vaccination of CBA/J mice, before pregnancy, with exosomes secreted by SRDCs pulsed in vitro with Toxoplasma gondii-derived antigens (TAg) induced a protective response in the pups. Indeed, vaccination resulted in the presence of significantly fewer cysts in pup brains. This protection was associated with strong humoral responses in the serum in vivo. We also observed cellular responses in vivo, with cell proliferation associated with the production of cytokines by the splenocytes. Thus, exosomes are nucleic acid-free vesicles able to induce immune responses correlated with protection against T. gondii infection in a congenital model. They are therefore a potentially useful tool for vaccination against infectious disease. PMID- 19186200 TI - Cost-effectiveness of childhood rotavirus vaccination in Taiwan. AB - OBJECTIVE: Rotavirus is the most common cause of severe diarrhea in children. Two rotavirus vaccines (RotaTeq and Rotarix) have been licensed in Taiwan. We have investigated whether routine infant immunization with either vaccine could be cost-effective in Taiwan. METHODS: We modeled specific disease outcomes including hospitalization, emergency department visits, hospital outpatient visits, physician office visits, and death. Cost-effectiveness was analyzed from the perspectives of the health care system and society. A decision tree was used to estimate the disease burden and costs based on data from published and unpublished sources. RESULTS: A routine rotavirus immunization program would prevent 146,470 (Rotarix) or 149,937 (RotaTeq) cases of rotavirus diarrhea per year, and would prevent 21,106 (Rotarix) and 23,057 (RotaTeq) serious cases (hospitalizations, emergency department visits, and death). At US$80 per dose for the Rotarix vaccine, the program would cost US$32.7 million, provided an increasing cost offset of US$19.8 million to the health care system with $135 per case averted. Threshold analysis identified a break-even price per dose of US$27 from the health care system perspective and US$41 from a societal perspective. At US$60.0 per dose of RotaTeq vaccine, the program would cost US$35.4 million and provide an increasing cost offset of US$22.5 million to the health care system, or US$150 per case averted. Threshold analysis identified a break-even price per dose of US$20.0 from the health care system perspective and $29 from the societal perspective. Greater costs of hospitalization and lower vaccine price could increase cost-effectiveness. CONCLUSIONS: Despite a higher burden of serious rotavirus disease than estimated previously, routine rotavirus vaccination would unlikely be cost-saving in Taiwan at present unless the price fell to US$41 (Rotarix) or US$29 (RotaTeq) per dose from societal perspective, respectively. Nonetheless, rotavirus immunization could reduce the substantial burden of short term morbidity due to rotavirus. PMID- 19186201 TI - Acute cerebellar ataxia in the Netherlands: a study on the association with vaccinations and varicella zoster infection. AB - AIM: Acute cerebellar ataxia (ACA, sudden onset of truncal ataxia and gait disturbances) usually follows a benign illness (25% varicella). It is also described after vaccination, like MMR and varicella zoster virus (VZV). We will establish incidence rates of (varicella related) ACA and assess the attributable risk of vaccination to ACA in the Netherlands. METHOD: Data on ACA in children, following infections, like varicella, and vaccinations, obtained from prospective, active pediatric surveillance and passive surveillance on adverse events following immunizations (AEFI) were compared with hospitalization data for ataxia. Capture-recapture (CRC) method was used to estimate the burden of ACA in the Netherlands. RESULTS: 45 children with ACA were included (44 and 1 reported by pediatric and AEFI surveillance respectively, 30 were hospitalized). Chickenpox preceded ACA in 15 cases, one case followed MMR. Of the hospitalization reports, 13 fulfilled the criteria for ACA. Using CRC the estimated number of hospitalized ACA cases was 42. For varicella related ACA, this estimate was 10, resulting in an incidence rate of 0.7:100,000 (95%CI 0.52 0.94, all cases) and 0.17:100,000 (95%CI 0.09-0.31, varicella related cases) for children under 15 years of age. CONCLUSION: The incidence rates were comparable with other studies. We found no association with MMR, but chickenpox was clearly related to ACA. According to age-specific seroprevalence data the incidence rate of ACA was 5:100,000 VZV infections for children up to 5 years, compared to an ACA-reporting rate of 0.15:100,000 doses VZV-vaccine. Therefore, uptake of VZV vaccine in the immunization programme will diminish the incidence rate of ACA. PMID- 19186202 TI - Comparative analysis of the immunogenicity of SARS-CoV nucleocapsid DNA vaccine administrated with different routes in mouse model. AB - The development of strategies to augment the immunogenicity of DNA vaccines is critical for improving their clinical utility. One such strategy involves using the different immune routes with DNA vaccines. In the present study, the immunogenicity of SARS-CoV nucleocapsid DNA vaccine, induced by using the current routine vaccination routes (intramuscularly, by electroporation, or orally using live-attenuated Salmonella typhimurium), was compared in mouse model. The comparison between the three vaccination routes indicated that immunization intramuscularly induced a moderate T cell response and antibody response. Mice administrated by electroporation induced the highest antibody response among the three immunization groups and a mid-level of cellular response. In contrast, the orally DNA vaccine evoked vigorous T cell response and a weak antibody production. These results indicated that the distinct types of immune responses were generated by the different routes of DNA immunization. In addition, our results also show that the delivery of DNA vaccines by electroporation and orally using live-attenuated Salmonella in vivo is an effective method to increase the immune responses. Further studies could be carried out using a combination strategy of both oral and electroporation immunizations to stimulate higher cellular and humoral immune responses. PMID- 19186203 TI - Long-term efficacy of plasma-derived and recombinant hepatitis B vaccines in a rural township of Central Taiwan. AB - AIMS: To assess the differences of long-term efficacy between plasma-derived and recombinant hepatitis B virus (HBV) vaccines and the effectiveness of catch-up vaccination in adolescents with undetectable anti-HBs. METHODS: Before 1992, infants born in Taiwan were immunized using plasma-derived HB vaccine, and thereafter, by using recombinant HB vaccine. From the only junior middle school of a rural township in central-southern Taiwan, 1788 (93.7%) students from five cross-sectional screenings, grouping into three birth cohorts (Group I: born during 1984-1986, II: 1986-1992 and III: 1992-1995), were enrolled for checking HBsAg, anti-HBs and anti-HBc. Students with undetectable HBsAg and anti-HBs underwent a booster dose (2.5ug) of recombinant HB vaccine (Engerix-B; GlaxoSmithKline, Rixensart, Belgium) and had anti-HBs re-checked 3 weeks later. Individuals who had remained undetectable for anti-HBs completed the other two doses of HB vaccines at 1 and 6 months later. RESULTS: The prevalence of HBsAg (11.4, 5.4 and 1.2%), anti-HBs (64.5, 44.1 and 36.0%) and anti-HBc (29.5, 12.5 and 4.4%) decreased from Group I to III (P<0.001 for trends). After a booster dose, the positive rates of anti-HBs increased up to 80.5% (16% increase) in Group I, 81.0% (36.9% increase) in Group II, and 94.4% (58.4% increase) in Group III. The percentages of anamnestic response increased with a trend (P<0.001). A total of 110 non-responders completed 3 doses of catch-up HB vaccination, but 3 cases (2.7%) of Group II, evoked primary vaccination response. CONCLUSION: Recombinant vaccine showed predominant disappearance rate (62.7%) of anti-HBs 12 15 years after vaccination, but provided better anamnestic response after a booster dose. It also showed high success rate (97.3%) in catch-up vaccination in adolescents. PMID- 19186204 TI - Appearance of skin lesions in cattle populations vaccinated against lumpy skin disease: statutory challenge. AB - The ultimate goal of a vaccine is to protect vaccinated animals against re exposure to the same pathogen and provide sterile immunity. However, a cutaneous clinical manifestation appeared, following re-exposure of cattle that had been vaccinated with the RM65 strain, to LSDV infection during an epidemic in 2006 2007. Four thousand six hundred and seven vaccinated cows entered the study after being re-exposed to LSDV infection. Of them, 513 (11%) presented lumps, and there was a marked difference between the proportions of dairy and feedlot animals that were affected: 146 out of 3517 and 367 out of 1090 (6.6 and 33.7%, respectively). This data suggests that the potency of the vaccine need to be re-assessed for beef cattle. PMID- 19186205 TI - Leadership in Immunization: the relevance to Japan of the U.S.A. experience of the Centers for Disease Control and Prevention (CDC) and the Advisory Committee on Immunization Practices (ACIP). AB - Immunization is one of the most effective public health interventions available. However, the difference of how recommendations are established influence the utilization of the vaccine and the number of patients with vaccine preventable diseases (VPDs). In this paper, we compared and highlighted the difference of the situation of the immunization program in Japan and the United States in terms of structure of policy setting system. Since the disease epidemiology clearly indicates that the US has better control of VPDs over Japan, we considered the advantage of development of a comparable vaccine policy setting system of the US in Japan. PMID- 19186206 TI - Rapid identification of herd effects with the introduction of serogroup C meningococcal conjugate vaccine in Ontario, Canada, 2000-2006. AB - In 2001, Canada's National Advisory Committee on Immunization endorsed a meningococcal serogroup C conjugate vaccine, which appears to provide durable serogroup-specific immunity while reducing nasopharyngeal carriage. With reference to direct and indirect effects on case occurrence, we sought to evaluate recent trends in the incidence of invasive meningococcal disease (IMD) in Ontario. Analyses included all IMD cases reported between 2000 and 2006 to the Ontario Central Public Health Laboratory. Poisson models incorporating terms for age, sex and seasonal oscillation identified a significant downward trend in disease occurrence, which was strongest in serogroup C cases and not evident when serogroup C strains were excluded from the analysis. Among age groups not targeted by the vaccine program serogroup C, IMD displayed a pattern of decreasing incidence that was not present in non-serogroup C disease. These apparent dramatic effects of conjugate C vaccine (both direct and indirect) may be important in the implementation and evaluation of vaccine policy in other jurisdictions. PMID- 19186207 TI - Utility of animal models for predicting human allergenicity. AB - The biochemical characterization of protein structures has led to a better understanding of allergens, their structure/function relationship, and can be very powerful in identifying protein sequences with significant structural similarity to known allergens. However, for scientists, regulators and food manufacturers there exists a need for acquiring additional data on potential allergenicity of proteins, particularly, biotechnology derived molecules in food products for which minimal or no prior human exposure information is available. Since human exposure testing, while direct, is unacceptable, understanding allergy in animals has been used to investigate the allergic response on a molecular level as well as test the potential in vivo allergenicity of food proteins. Rodents seem to be the most likely candidate for assessing allergenicity. For development of an animal test system for allergenicity characterization and testing, a number of criteria are required for qualification for a model of human allergy including acceptable immunization protocols, allergic response measurements, and for standardization and validation of materials and procedures. If an animal test system can minimally provide a basis for measuring the relative physiological response to known allergens, this should be enough to establish a model that produces a relative measure of potential allergenicity. Our article will consider development of an adequate animal model for allergenicity determination that can be validated as a tool in safety assessments. PMID- 19186208 TI - Increased susceptibility to kainic acid-induced seizures in Engrailed-2 knockout mice. AB - The En2 gene, coding for the homeobox-containing transcription factor Engrailed-2 (EN2), has been associated to autism spectrum disorder (ASD). Due to neuroanatomical and behavioral abnormalities, which partly resemble those observed in ASD patients, En2 knockout (En2(-/-)) mice have been proposed as a model for ASD. In the mouse embryo, En2 is involved in the specification of midbrain/hindbrain regions, being predominantly expressed in the developing cerebellum and ventral midbrain, and its expression is maintained in these structures until adulthood. Here we show that in the adult mouse brain, En2 mRNA is expressed also in the hippocampus and cerebral cortex. Hippocampal En2 mRNA content decreased after seizures induced by kainic acid (KA). This suggests that En2 might also influence the functioning of forebrain areas during adulthood and in response to seizures. Indeed, a reduced expression of parvalbumin and somatostatin was detected in the hippocampus of En2(-/-) mice as compared to wild type (WT) mice, indicating an altered GABAergic innervation of limbic circuits in En2(-/-) mice. In keeping with these results, En2(-/-) mice displayed an increased susceptibility to KA-induced seizures. KA (20 mg/kg) determined more severe and prolonged generalized seizures in En2(-/-) mice, when compared to WT animals. Seizures were accompanied by a widespread c-fos and c-jun mRNA induction in the brain of En2(-/-) but not WT mice. Long-term histopathological changes (CA1 cell loss, upregulation of neuropeptide Y) also occurred in the hippocampus of KA-treated En2(-/-) but not WT mice. These findings suggest that En2(-/-) mice might be used as a novel tool to study the link between epilepsy and ASD. PMID- 19186209 TI - The consolidation of neuroleptic therapy: Janssen, the discovery of haloperidol and its introduction into clinical practice. AB - The discovery of haloperidol at the end of the 1950s constitutes one of the greatest advances of 20th century psychiatry. This antipsychotic drug has their origin in the research process of central analgesic molecules derived from pethidine and methadone, carried out by the Belgian company Janssen Phamaceutica. After the synthesis of phenoperidine, numerous analogues of this compound were studied, and chemists at Janssen took the decision to substitute the propiophenone group for a butyrophenone group. One of these compounds went the R 1625, a stronger agent with specifically neuroleptic properties but lacking morphine-like activity. This substance was synthesized on the 11th February 1958 and received the generic name of haloperidol because of the two halogenated substitutes incorporated into the molecule. Clinical development of haloperidol was conducted, primarily, by psychiatric research team at the University of Liege that confirmed its efficacy in the treatment of various psychiatric disorders such as acute and chronic paranoid psychosis, mania, or chronic treatment resistant schizophrenia. Under the brand name Haldol((R)), haloperidol was licensed and marketed in Belgium in October 1959. The direct and differed consequences of its introduction into the psychiatric practice have been multiple, involving different areas of socio-sanitary reality. Moreover, haloperidol has contributed substantially to the development of biological psychiatry and currently neuroscience, because it made possible the development of new experimental models for predicting the effects of antipsychotics, and allowed the postulate of the firsts biological hypotheses about the schizophrenia etiology. Haloperidol has been included in the World Health Organisation's list of essential medicines. PMID- 19186210 TI - Synergistic interaction between the Ligusticum chuanxiong constituent butylidenephthalide and the nitric oxide donor sodium nitroprusside in relaxing rat isolated aorta. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Ligusticum chuanxiong Hort. (Umbelliferae), a traditional Chinese medicinal herb, is often prescribed together with nitric oxide donors for treating coronary heart diseases such as angina in China; however, studies concerning their pharmacological interaction are scarce. AIM OF THE STUDY: The objective of the present study was to examine the interaction between the Ligusticum chuanxiong major active constituent butylidenephthalide (BDPH) and the nitric oxide donor sodium nitroprusside (SNP) in vasorelaxation. MATERIALS AND METHODS: Vasorelaxation was examined in rat isolated aorta using an organ bath system. RESULTS: BDPH and SNP interacted synergistically under 9,11 dideoxy-9alpha,11alpha-methanoepoxyprostaglandin H2 (U-46619)-induced tone. This synergism became greater with increasing U-46619 concentrations where Ca2+ sensitization contributed more significantly, and less when U-46619 was replaced with phenylephrine where participation of Ca2+ sensitization was minimal. BDPH SNP synergism remained intact in the absence of external Ca2+, indicating that regulation of Ca2+ influx was not a requirement for the manifestation of this interaction. CONCLUSIONS: The present study demonstrated the synergistic relaxation between BDPH and SNP in rat isolated aorta. This interaction is related to an enhancement of the effectiveness of SNP in producing relaxation under tone induced mainly by Ca2+ sensitization. PMID- 19186212 TI - Post-training infusion of glutamate into the bed nucleus of the stria terminalis enhanced inhibitory avoidance memory: an effect involving norepinephrine. AB - This study examined an interaction between glutamate and norepinephrine in the bed nucleus of the stria terminalis (BNST) in modulating affective memory formation. Male Wistar rats with indwelling cannulae in the BNST were trained on a one-trial step-through inhibitory avoidance task and received pre- or post training intra-BNST infusion of glutamate, norepinephrine or their antagonists. Results of the 1-day test indicated that post-training intra-BNST infusion of DL 2-amino-5-phosphonovaleric acid (APV) impaired retention in a dose- and time dependent manner, while infusion of glutamate had an opposite effect. Co-infusion of 0.2microg glutamate and 0.02microg norepinephrine resulted in marked retention enhancement by summating non-apparent effects of the two drugs given at a sub enhancing dose. The amnesic effect of 5.0microg APV was ameliorated by 0.02microg norepinephrine, while the memory enhancing effect of 1.0microg glutamate was attenuated by 5.0microg propranolol. These findings suggest that training on an inhibitory avoidance task may alter glutamate neurotransmission, which by activating NMDA receptors releases norepinephrine to modulate memory formation via beta adrenoceptors in the BNST. PMID- 19186211 TI - Site-specific integration of retroviral DNA in human cells using fusion proteins consisting of human immunodeficiency virus type 1 integrase and the designed polydactyl zinc-finger protein E2C. AB - During the life cycle of retroviruses, establishment of a productive infection requires stable joining of a DNA copy of the viral RNA genome into host cell chromosomes. Retroviruses are thus promising vectors for the efficient and stable delivery of genes in therapeutic protocols. Integration of retroviral DNA is catalyzed by the viral enzyme integrase (IN), and one salient feature of retroviral DNA integration is its lack of specificity, as many chromosomal sites can serve as targets for integration. Despite the promise for success in the clinic, one major drawback of the retrovirus-based vector is that any unintended insertion events from the therapy can potentially lead to deleterious effects in patients, as demonstrated by the development of malignancies in both animal and human studies. One approach to directing integration into predetermined DNA sites is fusing IN to a sequence-specific DNA-binding protein, which results in a bias of integration near the recognition site of the fusion partner. Encouraging results have been generated in vitro and in vivo using fusion protein constructs of human immunodeficiency virus type 1 IN and E2C, a designed polydactyl zinc finger protein that specifically recognizes an 18-base pair DNA sequence. This review focuses on the method for preparing infectious virions containing the IN fusion proteins and on the quantitative PCR assays for determining integration site specificity. Efforts to engineer IN to recognize specific target DNA sequences within the genome may lead to development of effective retroviral vectors that can safely deliver gene-based therapeutics in a clinical setting. PMID- 19186213 TI - Environmental enrichment improves behavioral performance and auditory spatial representation of primary auditory cortical neurons in rat. AB - Environmental enrichment (EE) has an important role in brain plasticity. Early research has shown that EE increases the response strength of neurons in the auditory cortex, but it remains unknown whether EE can influence the process of spatial localization in the auditory system. In this study, we raised rats in enriched and standard conditions from postnatal day 10 to day 56. By testing behavioral tasks via auditory cues, we have shown that EE improved the number of correct scores, but decreased the reaction time and azimuth deviation in behavioral performance of sound-azimuth discrimination. By in vivo extracellular recording, we have shown that EE enhanced the directional sensitivity of neurons in the primary auditory cortex. For example, EE rats had a smaller spatial receptive field, sharper frequency tuning curve and directional selective curve of auditory neurons compared with normal rats. Our findings indicate that early exposure to EE increases directional sensitivity. These results provide an insight into developmental plasticity in the auditory system. PMID- 19186214 TI - Behavioral and neuronal attributes of short- and long-term habituation in the crab Chasmagnathus. AB - Investigations using invertebrate species have led to a considerable progress in our understanding of the mechanisms underlying learning and memory. In this review we describe the main behavioral and neuronal findings obtained by studying the habituation of the escape response to a visual danger stimulus in the crab Chasmagnathus granulatus. Massed training with brief intertrial intervals lead to a rapid reduction of the escape response that recovers after a short term. Conversely, few trials of spaced training renders a slower escape reduction that endures for many days. As predicted by Wagner's associative theory of habituation, long-term habituation in the crab proved to be determined by an association between the contextual environment of the training and the unconditioned stimulus. By performing intracellular recordings in the brain of the intact animal at the same time it was learning, we identified a group of neurons that remarkably reflects the short- and long-term behavioral changes. Thus, the visual memory abilities of crabs, their relatively simple and accessible nervous system, and the recording stability that can be achieved with their neurons provide an opportunity for uncovering neurophysiological and molecular events that occur in identifiable neurons during learning. PMID- 19186215 TI - Necrotic enteritis in chickens: a paradigm of enteric infection by Clostridium perfringens type A. AB - Withdrawal of antimicrobial growth promoters and ionophore coccidiostats has been accompanied by a resurgence in incidence of necrotic enteritis (NE), a severe Clostridium perfringens-induced disease which some consider the most clinically dramatic bacterial enteric disease of poultry. Lesions, in jejunum and ileum, are focal-to-confluent, often with a tightly adhered pseudomembrane, and hemorrhage is uncommon. The key risk factor for development of NE is an intestinal environment that favors growth of the organism. Birds on high energy, protein rich, wheat- or barley-based diets experience NE at a rate up to ten times greater than do birds on maize-based diets. Specific strains of type A cause NE, although only a few specific virulence attributes are known. The role of alpha toxin (CPA) has been called into question by the finding that an engineered CPA mutant retained full virulence in vivo, although the counterpoint to this is the finding that immunization with CPA toxoids provides substantial protection against NE. A recently described toxin, NetB, seems likely to be involved in pathogenesis of infection by most NE strains. Immunization with CPA, NetB, or other proteins, delivered by conventional means or vectored by recombinant attenuated Salmonella vectors may help the industry deal with NE. Future progress may be based in large part on genomic and proteomic analyses. PMID- 19186217 TI - Further comment on proton pump inhibitor failures. PMID- 19186216 TI - Do psychiatric comorbidities predict postoperative seizure outcome in temporal lobe epilepsy surgery? AB - Clinical and demographic presurgical variables may be associated with unfavorable postsurgical neurological outcome in patients with mesial temporal lobe epilepsy with hippocampal sclerosis (MTLE-HS). However, few reports include preoperative psychiatric disorders as a factor predictive of long-term postsurgical MTLE-HS neurological outcome. We used Engel's criteria to follow 186 postsurgical patients with MTLE-HS for an average of 6 years. DSM-IV criteria and psychiatric comorbidity criteria specific to epilepsy (interictal dysphoric disorder, postictal and interictal psychosis) were used to assess presurgical psychiatric disorders. Kaplan-Meier event-free survival and adjusted hazard ratios were estimated with unconditional logistic regression. Seventy-seven (41.4%) patients had a preoperative Axis I psychiatric diagnosis. Thirty-six patients had depression, 11 interictal dysphoric disorder, 14 interictal psychosis, 6 postictal psychosis, and 10 anxiety disorders. Twenty-three (12.4%) patients had Axis II personality disorders. Regarding seizure outcome, preoperative anxiety disorders (P=0.009) and personality disorders (P=0.003) were positively correlated with Engel class 1B (remaining auras) or higher. These findings emphasize the importance of presurgical psychiatric evaluation, counseling, and postsurgical follow-up of patients with epilepsy and psychiatric disorders. PMID- 19186219 TI - WITHDRAWN: Synthesis of magnetic nanoparticles for biological and water applications. PMID- 19186220 TI - Preparation and in vitro evaluation of mucoadhesion and permeation enhancement of thiolated chitosan-pHEMA core-shell nanoparticles. AB - The aim of the present work was to evaluate the in vitro mucoadhesion and permeation enhancement properties of thiolated chitosan (chitosan-glutathione) coated poly(hydroxyl ethyl methacrylate) nanoparticles. Core-shell nanoparticles were prepared by radical emulsion polymerization method initiated by cerium(IV) ammonium nitrate. Different molecular weights of chitosan were utilized for nanoparticles preparation. The physicochemical properties of nanoparticles were characterized by size, zeta potential, and thiol content. Incorporation of fluorescein isothiocyanate dextran (FD4, MW 4400 Da), which was used as the model macromolecule, was achieved by incubation method. The intestinal mucoadhesion and penetration enhancement properties of nanoparticles were investigated using excised rat jejunum. All nanoparticle systems showed mucoadhesion and improved apparent permeation coefficient (P(app)) of FD4. Nanoparticles prepared by thiolated chitosan with medium molecular weight revealed the most mucoadhesion and penetration enhancement properties. PMID- 19186221 TI - Polybacterial immunomodulator Respivax restores the inductive function of innate immunity in patients with recurrent respiratory infections. AB - Respivax (BulBio-NCIPD Ltd.) is an oral polybacterial immunomodulator intended for treatment and prevention of non-specific respiratory tract infections. We studied for the first time its effects on the inductive mechanisms of innate immunity, in the course of 3-month immunoprophylaxis of 25 patients with recurrent and chronic respiratory infections. The expression of pattern recognition receptors on peripheral blood (PB) monocytes and plymorphonuclear cells (PMNs), the antigen-presenting and co-stimulatory potential of peripheral blood monocytes and dendritic cells; and the stimulated Th1/Th2 cytokine production were determined by flow cytometry. As compared to healthy controls, patients were characterized with down-regulation of TLR2 and TLR4/CD14 complex on PB monocytes (p<0.01), decreased share of CD14+CD16+ DCs precursors (p<0.01), decreased CD86 expression on PB DCs (p<0.05) and a Th2 shift of cytokine profile. Respivax modulated differentially the surface expression of pattern-recognition receptors on PB monocytes, increasing TLR2 and CD14 without affecting TLR4 expression. Further on, Respivax enhanced the differentiation of mature CD86(high) dendritic cells (DCs). Importantly, Respivax promoted a Th1 shift of cytokine profile and restored the Th1/Th2 cytokine balance without pro inflammatory effects. Noteworthy, Th1/Th2 ratios in the patient's group correlated positively with the levels of TLR2 (R=0.5, p<0.001) and CD14 expression (R=0.4, p<0.05). We conclude that Respivax treatment restores the inductive function of innate immunity at three key levels: antigen recognition and presentation, co-stimulation of naive T cells, and Th1/Th2 balance. This results, at least in part, from a differential modulation effect on the expression of pathogen-recognition receptors. PMID- 19186222 TI - Pathogenesis of multi-organic failure in autoimmune diseases. AB - Multi-organic failure in the context of autoimmune diseases is a multi-factorial condition where different pathways concur to produce a global system breakdown. Some of these pathways include the coagulation, fibrinolysis, kinin and complement cascades which in normal conditions work together to provide a comprehensive response to injury. In pathologic conditions these regulatory mechanisms are replaced by positive feed-back loops. The common response pattern is the activation of the immune system via endothelium activation. Furthermore, these different plasma-driven mechanisms may induce standardised endothelial cell responses of which the most relevant are the activation of p38, JNK, NF-kbeta and IRF-3 pathways. In this paper we review the common points between these major pathways and how they become activated, contributing to a global clinical picture. We present two examples of apparently different clinical settings, caused by the same global dysfunction: the Macrophage Activation Syndrome and the iatrogenic "cytokine storm" triggered by the administration of anti-CD28 monoclonal antibody TGN1412 in a phase 1 trial. PMID- 19186223 TI - Critical analysis of rituximab-induced serological changes in connective tissue diseases. AB - A survey of PUBMED and EMBASE supplied 21 articles dealing with the effect of rituximab (RTX) on immunoglobulin (Ig) and autoantibodies (Abs) in nonorgan systemic autoimmune diseases, and another 12 articles were found by hand search. No statistics could be performed due to the lack of numerical data in the articles. RTX tended to diminish total Ig but still within the normal ranges, more for IgM than IgG and IgA. Rheumatoid factor level decreased by 30 to 60% 3-6 months after RTX, whereas anti-cyclic citrullinated peptide Ab titers declined modestly. In contrast, anti-DNA and anti-C1q Ab levels showed a marked decrease, whereas the other anti-extractable nuclear antigens Ab (anti-SSA, SSB, SM, RNP...) were stable. There are claims for an increase in the BAFF level by approximately 2.5 to 3-fold at 3-4 months, and a return to pre-treatment value at 8-12 months. RTX-induced changes in the serum bring about new insights into mechanisms of action. Therefore, more attention should be paid to such parameters in clinical trials. PMID- 19186224 TI - Human defensins as cancer biomarkers and antitumour molecules. AB - Human defensins, which are small cationic peptides produced by neutrophils and epithelial cells, form two genetically distinct alpha and beta subfamilies. They are involved in innate immunity through killing microbial pathogens or neutralizing bacterial toxins and in adaptive immunity by serving as chemoattractants and activators of immune cells. alpha-defensins are mainly packaged in neutrophil granules (HNP1, HNP2, HNP3) or secreted by intestinal Paneth cells (HD5, HD6), while beta-defensins are expressed in mucosa and epithelial cells. Using surface enhanced laser desorption/ionisation time-of flight (SELDI-TOF) mass spectrometry (MS), alpha-defensins were found to be expressed in a variety of human tumours, either in tumour cells or at their surface. HNP1-3 peptides are also secreted and their accumulation in biological fluids was proposed as a tumour biomarker. Conversely, beta-defensin-1 (HBD-1) is down-regulated in some tumour types in which it could behave as a tumour suppressor protein. Alpha-defensins promote tumour cell growth or, at higher concentration, provoke cell death. These peptides also inhibit angiogenesis, which, in addition to immunomodulation, indicates a complex role in tumour development. This review summarizes current knowledge of defensins to discuss their role in tumour growth, tumour monitoring and cancer treatment. PMID- 19186225 TI - Multidisciplinary approach to cancer care. Surgical clinics of North America. Foreword. PMID- 19186226 TI - Multidisciplinary approach to cancer care. Surgical clinics of North America. Preface. PMID- 19186227 TI - The molecular basis of cancer and the development of targeted therapy. AB - The sequencing of the human genome and the ability to rapidly identify genes and proteins, both normal and mutant, that are involved in tumorigenesis and malignant phenotypes, have changed the ability to understand malignant cells. Understanding and applying this information to the diagnosis and treatment of cancer are facilitated best with a multidisciplinary team. The cancer surgeon plays a pivotal role in this team. This article briefly summarizes: (1) the clinically relevant applications of molecular biology to the cancer surgeon, (2) the current understanding of the molecular basis for cancer, and (3) the current targeted agents and their clinical applications. PMID- 19186228 TI - Measuring outcomes in oncology treatment: the importance of patient-centered outcomes. AB - An important component of quality healthcare is that it be patient-centered with a focus on the patient, including his or her preferences, values, and beliefs. The goal of this article is to provide a broad overview of patient-centered outcomes in oncologic research. It starts with an introduction to the different types of patient-centered measures including patient satisfaction, decision regret, patient preference, and health-related quality of life. It then offers an overview of survey instrument design and selection. Finally, it provides examples of existing approaches to measurement and previously validated instruments for each type of patient-centered outcome. PMID- 19186229 TI - Surgical palliation: getting back to our roots. AB - This article provides an overview of the approach to patients who may benefit from palliative care. While the article's details lend themselves to the treatment of complications secondary to advanced malignancies, the data herein can also be extrapolated to other chronic, terminal diseases. Guidelines for patient selection are discussed, using currently available outcomes data as a platform for the critical decision making process. Suggestions for a multidisciplinary team approach are offered, using the palliative triangle as the ideal model of communication and cooperation. Finally, methods for measuring success are detailed, along with proposals for how to better equip the surgeons of tomorrow with the knowledge and experience needed to tackle these difficult and intimate problems. PMID- 19186230 TI - Multimodal treatment for head and neck cancer. AB - Head and neck cancers are relatively less common tumors, but with complex anatomic and physiologic relationships to the structures from which they arise. Multimodal management is required for advanced stage disease, while single modality treatment is usually sufficient for early lesions. Treatment paradigms have shifted toward more functional preservation of speech and swallowing, when possible. Increased use of radiation, systemic/targeted therapies and function preserving surgical approaches have allowed for organ preservation without compromising oncologic outcomes in properly selected patients. PMID- 19186231 TI - The role of minimally invasive treatments in surgical oncology. AB - This article reviews the use of minimally invasive surgical and endoscopic techniques in the field of surgical oncology. It reviews the indications and techniques of the use of minimally invasive surgery for several oncologic indications in general surgery. In particular, it reviews the currently published literature discussing the oncologic outcomes of these techniques. PMID- 19186232 TI - Multidisciplinary approach to esophageal and gastric cancer. AB - The incidence of esophageal and gastric malignancies has increased over the last decade. Historically, surgery has been considered the best treatment for these cancers. However, long-term survival after surgery is fair at best, because of the tendency of disease to recur locally and distantly. Presently, the management of these cancers involves surgery, chemotherapy, and radiation therapy. This article discusses various treatment strategies that employ these modalities either alone or in combination, in an attempt to improve survival rates for patients who have gastroesophageal malignancies. PMID- 19186233 TI - Liver-directed treatment modalities for primary and secondary hepatic tumors. AB - Colorectal cancer liver metastases and hepatocellular carcinoma remain significant health problems in the United States and worldwide. Although surgical resection is often the treatment of choice, patient comorbidities or disease extent may preclude this option. Alternative approaches to primary and secondary hepatic malignancies have been developed, and their impact on disease control has been the subject of much recent study. These therapies can be administered alone but can also be effective when used in combination, or with other chemotherapeutic regimens. This article reviews the different techniques of liver directed therapy and the available literature on short- and long-term outcomes. PMID- 19186234 TI - Multidisciplinary approach to tumors of the pancreas and biliary tree. AB - Tumors of the pancreas and biliary tree remain formidable challenges to patients and clinicians. These tumors elude early detection, rapidly spread locally and systemically, and frequently recur despite apparently complete resection. Cystic tumors of the pancreas, however, may represent a subset of patients who do not uniformly require aggressive resection, and a thoughtful, evidence-based approach to work-up allows for the rational application of surgical therapy. Increasing evidence supports treating patients who have pancreaticobiliary disease in a multidisciplinary setting. PMID- 19186235 TI - Multidisciplinary care for patients with breast cancer. AB - The care of patients with breast cancer has become increasingly complex with advancements in diagnostic modalities, surgical approaches, and adjuvant treatments. A multidisciplinary approach to breast cancer care is essential to the successful integration of available therapies. This article addresses the key components of multidisciplinary breast cancer care, with a special emphasis on new and emerging approaches over the past 10 years in the fields of diagnostics, surgery, radiation, medical oncology, and plastic surgery. PMID- 19186236 TI - The multidisciplinary management of rectal cancer. AB - Advancements have been made in multiple aspects of diagnostic and therapeutic approaches to rectal cancer. These advances include clinical staging such as endorectal ultrasound and pelvic MRI, surgical approaches such as transanal excision, and adjuvant treatments such as new chemotherapeutic agents and refined radiotherapy techniques. Optimal patient outcomes depend on multidisciplinary involvement for tailored therapy. The successful management of rectal cancer requires a multidisciplinary approach, with treatment decisions based on precise patient evaluations by a group of clinicians, including surgeons, gastroenterologists, medical and radiation oncologists, radiologists, and pathologists. The accurate identification of patients who are candidates for combined modality treatment is particularly essential to optimize outcomes. Technical and technologic advances have led to the availability of a wide range of surgical approaches for managing rectal cancer. Concomitantly, similar critical developments and refinements have also occurred in the administration of radiation and chemotherapeutic agents. This article provides an overview of the multimodal treatment of patients who have rectal cancer, with a focus on staging, surgical techniques, and the application of chemotherapy or radiation in the adjuvant and neoadjuvant settings. PMID- 19186238 TI - Soft tissue sarcomas: current management and future directions. AB - This article reviews the current state of diagnosis and treatment of soft tissue sarcomas. Etiology, staging, imaging, tissue sampling, and current treatment are all reviewed using updated references. Current standards for surgical treatment are emphasized and the future directions of treatment addressed. PMID- 19186239 TI - The surgical and systemic management of neuroendocrine tumors of the pancreas. AB - Neuroendocrine tumors of the pancreas comprise a class of rare tumors that can be associated with symptoms of hormone overproduction. Five distinct clinical endocrinopathies are associated with neuroendocrine tumors; however, most of these tumors remain asymptomatic and follow an indolent course. Complete surgical resection offers the only hope for cure, but understanding the basic biology of the tumors has advanced the medical management in metastatic disease. Surgical resection of hepatic metastases offers survival advantage and should be performed when feasible. Although hepatic artery embolization is currently the preferred mode of nonsurgical palliation for pain and hormonal symptoms, other modalities may play a role in metastatic disease. PMID- 19186237 TI - Multidisciplinary treatment of gastrointestinal stromal tumors. AB - Gastrointestinal stromal tumor (GIST) has been recognized as a unique tumor only in the last decade. Although rare as a clinical entity, there is much interest in the pathology and treatment because the KIT protooncogene mutation common to most GISTs can be inhibited by imatinib mesylate. Diagnosing and treating GIST requires a multidisciplinary approach, given the combination of pathologic and radiographic evaluation, surgical treatment, and oncologic care required to successfully treat patients with GIST. PMID- 19186240 TI - Multidisciplinary treatment of primary melanoma. AB - This article covers the multidisciplinary treatment of primary melanoma. Excision margins and the need for sentinel lymphadenectomy are mainly dictated by the Breslow thickness although exceptions to this dictum do exist. Interferon is the only FDA approved adjuvant therapy for high risk melanoma although its overall survival benefit is minimal. Trials examining different doses or duration of interferon therapy have not demonstrated any promising survival data so far. There have been several randomized vaccine trials for melanoma but none have shown an overall survival benefit. Research into T-cell regulation continues and will hopefully bring promise for the future of melanoma treatment. PMID- 19186241 TI - Preface. Mouse models of developmental genetic disease. PMID- 19186242 TI - Modeling neural tube defects in the mouse. AB - Neural tube defects (NTDs) are among the most common structural birth defects observed in humans. Mouse models provide an excellent experimental system to study the underlying causes of NTDs. These models not only allow for identification of the genes required for neurulation, they provide tractable systems for uncovering the developmental, pathological and molecular mechanisms underlying NTDs. In addition, mouse models are essential for elucidating the mechanisms of gene-environment and gene-gene interactions that contribute to the multifactorial inheritance of NTDs. In some cases these studies have led to development of approaches to prevent NTDs and provide an understanding of the underlying molecular mechanism of these therapies prevent NTDs. PMID- 19186243 TI - The etiopathogenesis of cleft lip and cleft palate: usefulness and caveats of mouse models. AB - Cleft lip and cleft palate are frequent human congenital malformations with a complex multifactorial etiology. These orofacial clefts can occur as part of a syndrome involving multiple organs or as isolated clefts without other detectable defects. Both forms of clefting constitute a heavy burden to the affected individuals and their next of kin. Human and mouse facial traits are utterly dissimilar. However, embryonic development of the lip and palate are strikingly similar in both species, making the mouse a model of choice to study their normal and abnormal development. Human epidemiological and genetic studies are clearly important for understanding the etiology of lip and palate clefting. However, our current knowledge about the etiopathogenesis of these malformations has mainly been gathered throughout the years from mouse models, including those with mutagen-, teratogen- and targeted mutation-induced clefts as well as from mice with spontaneous clefts. This review provides a comprehensive description of the numerous mouse models for cleft lip and/or cleft palate. Despite a few weak points, these models have revealed a high order of molecular complexity as well as the stringent spatiotemporal regulations and interactions between key factors which govern the development of these orofacial structures. PMID- 19186244 TI - Murine models of holoprosencephaly. AB - Holoprosencephaly (HPE), the most common developmental defect of the forebrain and midface, is caused by a failure to delineate the midline in these structures. Both genetic and environmental etiologies exist for HPE, and clinical presentation is highly variable. HPE occurs in sporadic and inherited forms, and even HPE in pedigrees is characterized by incomplete penetrance and variable expressivity. Heterozygous mutations in eight different genes have been identified in human HPE, and disruption of Sonic hedgehog expression and/or signaling in the rostroventral region of the embryo is a major common effect of these mutations. An understanding of the mechanisms whereby genetic defects and teratogenic exposures become manifest as developmental anomalies of varying severity requires experimental models that accurately reproduce the spectrum of defects seen in human HPE. The mouse has emerged as such a model, because of its ease of genetic manipulation and similarity to humans in development of the forebrain and face. HPE is generally observed in mice homozygous for mutations in orthologs of human HPE genes though, unlike humans, rarely in mice with heterozygous mutations. Moreover, reverse genetics in the mouse has provided a wealth of new candidate human HPE genes. Construction of hypomorphic alleles, interbreeding to produce double mutants, and analysis of these mutations on different genetic backgrounds has generated multiple models of HPE and begun to provide insight into the conundrum of the HPE spectrum. Here, we review forebrain development with an emphasis on the pathways known to be defective in HPE and describe the strengths and weaknesses of various murine models of HPE. PMID- 19186245 TI - Mouse models of congenital cardiovascular disease. AB - Congenital heart defects occur in nearly 1% of human live births and many are lethal if not surgically repaired. In addition, the genetic contribution to congenital or acquired cardiovascular diseases that are silent at birth, but progress to cause significant disease in later life is being increasingly appreciated. Heart development and structure are highly conserved between mouse and human. The discoveries that are being made in this model system are highly relevant to understanding the pathogenesis of human heart defects whether they occus in isolation, or in the context of a syndrome. Many of the genes required for cardiovascular development were discovered fortuitously when early lethality or structural defects were observed in mouse mutants generated for other purposes, and relevant genes continue to be defined in this manner. Candidate genes for this process are being identified by their roles other species, or by their expression in pertinent tissues in mice. In this review, I will briefly summarize heart development as currently understood in the mouse, and then discuss how complementary studies in mouse and human have identified genes and pathways that are critical for normal cardiovascular development, and for maintaining the structure and function of this organ system throughout life. PMID- 19186246 TI - Modeling ciliopathies: Primary cilia in development and disease. AB - Primary (nonmotile) cilia are currently enjoying a renaissance in light of novel ascribed functions ranging from mechanosensory to signal transduction. Their importance for key developmental pathways such as Sonic Hedgehog (Shh) and Wnt is beginning to emerge. The function of nodal cilia, for example, is vital for breaking early embryonic symmetry, Shh signaling is important for tissue morphogenesis and successful Wnt signaling for organ growth and differentiation. When ciliary function is perturbed, photoreceptors may die, kidney tubules develop cysts, limb digits multiply and brains form improperly. The etiology of several uncommon disorders has recently been associated with cilia dysfunction. The causative genes are often similar and their cognate proteins certainly share cellular locations and/or pathways. Animal models of ciliary gene ablation such as Ift88, Kif3a, and Bbs have been invaluable for understanding the broad function of the cilium. Herein, we describe the wealth of information derived from the study of the ciliopathies and their animal models. PMID- 19186248 TI - Fraying at the edge mouse models of diseases resulting from defects at the nuclear periphery. AB - Eukaryotic cells compartmentalize their genetic material within the nucleus. The boundary separating the genetic material from the cytoplasm is the nuclear envelope (NE) and lamina. Historically, the NE was perceived as functioning primarily as a barrier regulating the entry and exit of macromolecules between the nucleus and cytoplasm via the nuclear pore complexes (NPCs) that traverse the nuclear membranes. However, recent findings have caused a fundamental reassessment with regard to NE and lamina functions. Evidence now points to the NE and lamina functioning as a "hub" in regulating and perhaps integrating critical cellular functions that include chromatin organization, transcriptional regulation, mechanical integrity of the cell, signaling pathways, as well as acting as a key component of the cytoskeleton. Such an integral role for the nuclear boundary has emerged from increased interest into the functions of the NE/lamina, which has been largely stimulated by the discovery that some 24 different diseases and anomalies are caused by defects in proteins of the NE and lamina. PMID- 19186247 TI - Mouse models of polycystic kidney disease. AB - Polycystic kidney disease (PKD) is a diverse group of human monogenic lethal conditions inherited as autosomal dominant (AD) or recessive (AR) traits. Recent development of genetically engineered mouse models of ADPKD, ARPKD, and nephronophthisis/medullary cystic disease (NPHP) are providing additional insights into the molecular mechanisms governing of these disease processes as well as the developmental differentiation of the normal kidney. Genotypic and phenotypic mouse models are discussed and provide evidence for the fundamental involvement of cell-matrix, cell-cell, and primary cilia-lumen interactions, as well as epithelial proliferation, apoptosis, and polarization. Structure/function relationships between the PKD1, PKD2, PKHD1, and NPHP genes and proteins support the notion of a regulatory multiprotein cystic complex with a mechanosensory function that integrates signals from the extracellular environment. The plethora of intracellular signaling cascades that can impact renal cystic development suggest an exquisitely sensitive requirement for integrated downstream transduction and provide potential targets for therapeutic intervention. Appropriate genocopy models that faithfully recapitulate the phenotypic characteristics of the disease will be invaluable tools to analyze the effects of modifier genes and small molecule inhibitor therapies. PMID- 19186249 TI - Mouse models for human hereditary deafness. AB - Hearing impairment is a frequent condition in humans. Identification of the causative genes for the early onset forms of isolated deafness began 15 years ago and has been very fruitful. To date, approximately 50 causative genes have been identified. Yet, limited information regarding the underlying pathogenic mechanisms can be derived from hearing tests in deaf patients. This chapter describes the success of mouse models in the elucidation of some pathophysiological processes in the auditory sensory organ, the cochlea. These models have revealed a variety of defective structures and functions at the origin of deafness genetic forms. This is illustrated by three different examples: (1) the DFNB9 deafness form, a synaptopathy of the cochlear sensory cells where otoferlin is defective; (2) the Usher syndrome, in which deafness is related to abnormal development of the hair bundle, the mechanoreceptive structure of the sensory cells to sound; (3) the DFNB1 deafness form, which is the most common form of inherited deafness in Caucasian populations, mainly caused by connexin-26 defects that alter gap junction communication between nonsensory cochlear cells. PMID- 19186250 TI - The value of mammalian models for duchenne muscular dystrophy in developing therapeutic strategies. AB - Duchenne muscular dystrophy (DMD) is the most common form of muscular dystrophy. There is no effective treatment and patients typically die in approximately the third decade. DMD is an X-linked recessive disease caused by mutations in the dystrophin gene. There are three mammalian models of DMD that have been used to understand better the pathogenesis of disease and develop therapeutic strategies. The mdx mouse is the most widely used model of DMD that displays some features of muscle degeneration, but the pathogenesis of disease is comparatively mild. The severity of disease in mice lacking both dystrophin and utrophin is similar to DMD, but one has to account for the discrete functions of utrophin. Canine X linked muscular dystrophy (cxmd) is the best representation of DMD, but the phenotype of the most widely used golden retriever (GRMD) model is variable, making functional endpoints difficult to ascertain. Although each mammalian model has its limitations, together they have been essential for the development of several treatment strategies for DMD that target dystrophin replacement, disease progression, and muscle regeneration. PMID- 19186251 TI - Stability and design of alpha-helical peptides. PMID- 19186252 TI - Folding and wrapping soluble proteins exploring the molecular basis of cooperativity and aggregation. PMID- 19186253 TI - Rescuing proteins of low kinetic stability by chaperones and natural ligands phenylketonuria, a case study. PMID- 19186254 TI - The endoplasmic reticulum crossroads for newly synthesized polypeptide chains. PMID- 19186255 TI - All-atom protein folding with free-energy forcefields. PMID- 19186256 TI - Folding considerations for therapeutic protein formulations. PMID- 19186257 TI - Melamine and food safety in China. PMID- 19186258 TI - Over-the-counter medicines: in whose best interests? PMID- 19186259 TI - Appointment of PEPFAR head should be merit based. PMID- 19186260 TI - The global financial crisis: an acute threat to health. PMID- 19186261 TI - Dietary fibre: an agreed definition. PMID- 19186262 TI - Ian Gilmore: president of the Royal College of Physicians. PMID- 19186263 TI - N-3 polyunsaturated fatty acids and statins in heart failure. PMID- 19186264 TI - N-3 polyunsaturated fatty acids and statins in heart failure. PMID- 19186265 TI - N-3 polyunsaturated fatty acids and statins in heart failure. PMID- 19186266 TI - N-3 polyunsaturated fatty acids and statins in heart failure. PMID- 19186267 TI - N-3 polyunsaturated fatty acids and statins in heart failure. PMID- 19186269 TI - N-3 polyunsaturated fatty acids and statins in heart failure. PMID- 19186270 TI - Sure start in England. PMID- 19186271 TI - Sure start in England. PMID- 19186272 TI - Clinical use of exhaled nitric oxide measurements. PMID- 19186274 TI - Principles for allocation of scarce medical interventions. AB - Allocation of very scarce medical interventions such as organs and vaccines is a persistent ethical challenge. We evaluate eight simple allocation principles that can be classified into four categories: treating people equally, favouring the worst-off, maximising total benefits, and promoting and rewarding social usefulness. No single principle is sufficient to incorporate all morally relevant considerations and therefore individual principles must be combined into multiprinciple allocation systems. We evaluate three systems: the United Network for Organ Sharing points systems, quality-adjusted life-years, and disability adjusted life-years. We recommend an alternative system-the complete lives system which prioritises younger people who have not yet lived a complete life, and also incorporates prognosis, save the most lives, lottery, and instrumental value principles. PMID- 19186275 TI - A fisherman who could not row. PMID- 19186276 TI - Clinics in Chest Medicine. Preface. PMID- 19186277 TI - Approach to the patient with sepsis. AB - This article reviews the current understanding of sepsis, severe sepsis, and septic shock. The article details definitions and epidemiology pertinent to the sepsis syndrome. A brief discussion of mechanisms of disease is followed a description of organ-specific failures related to sepsis. A concise review of the latest treatment options for each organ dysfunction is provided. PMID- 19186278 TI - Adrenal insufficiency in septic shock. AB - During disease states, the endocrine axes exhibit different levels of activity according to the severity of illness. These various alterations have been widely investigated. Indeed, evidence indicates that the anterior pituitary is dysfunctional in these states, especially when multiple organ dysfunction syndrome is present, impacting both adrenal and thyroid glands, but also secretion of estrogen, growth hormone, insulin-like growth factor-1, and prolactin. In a majority of these cases, substitutive treatment is not obviously beneficial, inappropriate secretions being considered as adaptive responses to stressful events. The hypothalamic-pituitaryadrenal axis appears to play the most important role in the regulation of inflammation during septic shock. Many factors modulate this axis. Some are well known. Others, such as vasopressin and apelin, are newly ascribed. Therapeutic issues in critically ill patients still remain controversial and are ardently debated, especially with regard to the needs and practical use of corticosteroids in septic shock. This article focuses on actual knowledge, mechanisms, definitions, and therapeutic recommendations, as well as on areas of uncertainty relative to adrenal gland insufficiency in septic shock. PMID- 19186279 TI - Acute kidney injury in the intensive care unit. AB - Acute kidney injury (AKI) is common among critically ill patients and results in increased mortality in this population. This review focuses on the diagnosis and management of AKI. The authors first explore new aspects of diagnosis, including new criteria that take into account even modest changes in renal function, and the development of novel biomarkers to allow earlier identification and better differential diagnosis of AKI. The authors also explore the available data on choice of dialysis modality and dialysis dose for the treatment of AKI, as well as the breakthrough development of the bioartificial kidney. Last, the authors review co-interventions that may have relevance to prognosis of AKI, such as intensive insulin therapy and the use of erythropoietin. PMID- 19186280 TI - Intra-abdominal hypertension: evolving concepts. AB - This article focuses primarily on the recent literature on abdominal compartment syndrome (ACS) and the definitions and recommendations published by the World Society for the Abdominal Compartment Syndrome. The definitions regarding increased intra-abdominal pressure (IAP) are listed and are followed by an overview of the different mechanisms of organ dysfunction associated with intra abdominal hypertension (IAH). Measurement techniques for IAP are discussed, as are recommendations for organ function support and options for treatment in patients who have IAH. ACS was first described in surgical patients who had abdominal trauma, bleeding, or infection; but recently, ACS has been described in patients who have other pathologies. This article intends to provide critical care physicians with a clear insight into the current state of knowledge regarding IAH and ACS. PMID- 19186281 TI - ICU management of acute liver failure. AB - Survival of patients presenting with acute liver failure (ALF) has improved because of earlier disease recognition, better understanding of pathophysiology of various insults leading to ALF, and advances in supportive measures including a team approach, better ICU care, and liver transplantation. This article focuses on patient management and evaluation that takes place in the ICU for patients who have acute liver injury. An organized team approach to decision making about critical care delivered during this period of time is important for achieving a good patient outcome. PMID- 19186282 TI - Obstetric disorders in the ICU. AB - Obstetric disorders account for 55% to 80% of admissions to the intensive care unit in the obstetric population. Medical conditions are emerging as the leading cause of maternal mortality, partly because of marked improvement in surgical and obstetric care in the developed world. The rise in maternal mortality related to medical conditions can be explained by multiple factors: improved medical care, women with chronic illnesses reaching childbearing years, older age at time of first pregnancy, improved reproductive technologies, and severe medical conditions exacerbated by the physiologic changes of pregnancy. This article reviews obstetric disorders leading to intensive care unit admissions. PMID- 19186283 TI - Critical care management of subarachnoid hemorrhage and ischemic stroke. AB - This article addresses current critical care management strategies for patients with aneurysmal subarachnoid hemorrhage (SAH) and ischemic stroke. The chronology of typical SAH care is discussed, with an emphasis on the treatment of poor-grade SAH and on current monitoring techniques. Acute diagnosis and treatment of the life-threatening ischemic strokes is reviewed, emphasizing the care of patients with acute basilar artery occlusion and the malignant middle cerebral artery syndrome. Up-to-date estimates of the long-term outcomes of these syndromes are reviewed. PMID- 19186284 TI - Coagulation disorders in the ICU. AB - Disorders of hemostasis and thrombosis are frequently encountered in the ICU setting. Understanding the relevance of laboratory findings is essential in providing appropriate therapy. Various blood products and hemostatic agents are available to assist in the control of bleeding, and several different classes of anticoagulants are now available for use. Appropriate use of these agents maximizes therapeutic effect while minimizing complications. Use of fresh frozen plasma, cryoprecipitate, and other hemostatic agents should generally be reserved for those who have active bleeding, those undergoing invasive procedures, and those at high risk for bleeding because of their underlying diagnoses or because of associated hematologic derangements. PMID- 19186285 TI - Sedation and analgesia for the mechanically ventilated patient. AB - Mechanically ventilated patients in the intensive care unit routinely require sedative and analgesic medications to manage pain and anxiety. These medications may have unpredictable effects with long-term use. Strategies that may help to improve patient outcomes include thoughtful selection of medications, use of objective sedation and pain scales, and implementation of protocolized sedation. PMID- 19186286 TI - Short- and long-term cognitive outcomes in intensive care unit survivors. AB - Evidence increasingly suggests that cognitive impairment is common in intensive care unit survivors, although the nature, severity, and natural history remain unclear. Although the cognitive impairments improve over time in some individuals, they often fail to completely abate. While the functional correlates of these impairments are under-studied, cognitive impairments may adversely impact quality of life, ability to return to work or to work at previously established levels, and ability to function effectively in emotional and interpersonal domains. The potential etiologies of cognitive impairments in intensive care unit survivors are not fully understood and are likely heterogeneous and vary widely across patients. The contributions of these many factors may be particularly significant in patients with pre-existing vulnerabilities for the development of cognitive impairments, such as mild cognitive impairment, dementia, prior traumatic brain injury, or other comorbid disorders, as well as predisposing genetic factors. PMID- 19186287 TI - Critical care outcomes in the hematologic transplant recipient. AB - Critical illness is a dreaded complication in recipients of hematologic stem cell transplantation, with poor survival described in a number of early series. The perception of futility may impact the approach to therapy. Over the last 20 years, there have been significant advances in the application of intensive care, as well as changes in the management of patients before, during, and after transplantation. These advances and changes may have an impact on this traditionally poor survival. This article discusses the outcomes of critical illness in bone marrow and stem cell transplant recipients, and gives possible reasons for the apparent improvement in outcomes seen in more recent series. PMID- 19186288 TI - Measurement of quality and assurance of safety in the critically ill. AB - The global health care community has worked tirelessly for nearly a decade to make medical care safer for patients, but it still has limited ability to evaluate whether safety has improved. While there is a universal push to measure safety outcomes, the main barrier has been poor investment in the basic science of patient safety. This science would allow us to comprehend the causes of harm, design and pilot test interventions to reduce harm, and robustly evaluate their impact. This article describes several dilemmas in measuring patient safety, outlines a conceptual model and presents a framework for measuring patient safety, and offers future directions. Future research should seek to create a scientifically sound and feasible safety scorecard and improve performance. PMID- 19186289 TI - End-of-life decision making in the ICU. AB - A large proportion of deaths, particularly in the developed world, follows admission to an ICU. Therefore, end-of life decision making is an essential facet of critical care practice. For intensivists, managing death in the critically ill has become a key professional skill. They must be thoroughly familiar with the ethical framework that guides end-of-life decision making. Decisions should generally be made collaboratively by clinicians partnering with patients' families. Treatment choices should be crafted to meet specific, achievable goals. A rational, empathic approach to working with families should encourage appropriate, mutually satisfactory outcomes. PMID- 19186291 TI - [Relations between respiratory medicine and the pharmaceutical industry]. PMID- 19186293 TI - [Interleukin 8 concentrations in donor bronchoalveolar lavage: impact on primary graft failure in double lung transplant]. AB - BACKGROUND AND OBJECTIVE: The purpose of this study was to determine concentrations of interleukin 8 (IL-8) in the bronchoalveolar lavage (BAL) fluid from donor lungs and assess the role of IL-8 levels in the development of primary graft failure. PATIENTS AND METHOD: Twenty patients who received a double lung transplant were studied. A series of data, including BAL fluid concentrations of IL-8, were collected for the donors. Data collected for the recipients included arterial blood gases after 6, 24, and 48 hours, and intubation time. Patients with a ratio of PaO(2) to the fraction of inspired oxygen (FiO(2)) of less than 300 during the first 48 hours were diagnosed with primary graft failure. IL-8 levels were determined by enzyme-linked immunosorbent assay. Associations between the donor variables and IL-8 concentrations were evaluated using the Spearman rank correlation coefficient (rho) and the Mann-Whitney test for categorical and continuous variables, respectively. Logistic regression was used for multivariate analysis. RESULTS: Fifteen of the 20 donors were men. The cause of brain death was trauma in 9 donors, 7 were smokers, 13 required inotropic support, and pathogens were isolated in the BAL fluid of 18. The median age was 35 years (interquartile range [IQR], 23.5-51.25y), the median ventilation time was 1 day (IQR, 1-2d), the median PaO(2)/FiO(2) was 459.5 (IQR, 427-510.25), and the median IL-8 concentration in BAL fluid was 49.01ng/L (IQR, 7.86-94.05ng/mL). Ten of the recipients were men and the median age was 48.43 years (IQR, 25.4-56.81y). The median ischemic time was 210 minutes (IQR, 176.25-228.75 min) for the first lung and 300 minutes (IQR, 273.75-333.73 min) for the second lung. The median PaO(2)/FiO(2) ratio for the implant at 6, 14, and 48 hours was 329 (IQR, 190.25 435), 363.5 (IQR, 249-434.75), and 370.5 (IQR, 243.25-418.25), respectively. The median intubation time was 39.5 hours (IQR, 19.25-68.5h) and the correlation with IL-8 values was positive: higher IL-8 concentrations in BAL fluid correlated with longer ventilation times (Spearman rank correlation, P=.007; rho=0.583). Five patients developed primary graft failure; IL-8 concentrations were significantly higher in these patients than in those whose grafts did not fail (Mann-Whitney test, P=.003). CONCLUSION: High IL-8 concentrations in donor BAL fluid lead to longer ventilation time in the recipients and favor the development of primary graft failure after lung transplant. PMID- 19186292 TI - [Phenotypic characterization and course of chronic obstructive pulmonary disease in the PAC-COPD Study: design and methods]. AB - BACKGROUND AND OBJECTIVES: The Phenotype and Course of Chronic Obstructive Pulmonary Disease (PAC-COPD) study aims to improve our understanding of the phenotypic heterogeneity of this disease and the extent to which this heterogeneity is related to clinical course. The main objectives are a) to characterize the phenotypic variability in first-time hospitalizations for exacerbation of COPD and to propose a classification into subtypes and b) to ascertain the association between the defined subtypes and the clinical and functional course of COPD. PATIENTS AND METHODS: This is a cross-sectional and cohort study of 342 patients with COPD from 9 tertiary hospitals in 3 autonomous communities. The minimum follow-up period is 5 years. The main variables of interest are respiratory symptoms, smoking, alcohol use, physical activity, use of health care services, medical care, treatment received, activities of daily living, comorbid conditions, sleepiness, anxiety and depression, quality of life, forced spirometry and bronchodilation tests, lung volume and inspiratory capacity measured by body plethysmography, carbon monoxide diffusing capacity, baseline arterial blood gas values, respiratory and peripheral muscle function, electrocardiogram, body weight and composition measured by bioelectric impedance, chest radiograph, skin prick test, capacity for exercise measured in the 6-minute walk test and cardiopulmonary exercise test, induced sputum (for quantitative microbiological culture and determination of inflammatory markers), nighttime pulse oximetry, chest computed tomography scan, and echocardiography. Serum and plasma samples are also taken to measure levels of inflammatory markers and oxidative stress, for genetic analysis, and for other possible measurements that might be required in the future. The statistical analysis combines factor analysis and survival models such as Cox regression analysis. This project will enable us to reconsider the definition and classification of COPD and to better understand the factors associated with its natural history. PMID- 19186294 TI - [Program for the prevention of smoking in secondary school students]. AB - BACKGROUND AND OBJECTIVE: School smoking prevention programs have never yielded the expected results. The aim of this study was to analyze the efficacy of an intensive smoking prevention program created by the educational community in which it was to be applied. POPULATION AND METHOD: A 3-year smoking prevention program was carried out among the students of Fuentesauco Secondary School in Zamora, Spain. The Babilafuente Secondary School in Salamanca, Spain was the control group. The program included both prevention and treatment activities. The former were carried out in the school, in out-of-school situations, and in the community. The questionnaire of the European Smoking Prevention Framework Approach was used. RESULTS: A total of 417 students aged 12 to 17 years participated in the study. Of these, 54.4% belonged to the intervention group and 45.6% to the control group. Smokers represented 36.7% of the population. After the intervention smokers represented 40.1% of the Fuentesauco students compared with 46.1% of the Babilafuente students, though the difference was not statistically significant. With respect to the cognitive determinants of smoking behavior, after the intervention significant differences in favor of the intervention group were only observed in the subjects' perception of the behavior of their siblings, peers, and teachers. CONCLUSIONS: The use of smoking prevention programs in schools should be reconsidered, and their evaluation should be based on educational rather than clinical criteria. Proposed changes in the program include decreasing its intensity, starting with students of an earlier age and seeking greater involvement of parents. PMID- 19186295 TI - [Cultural adaptation into Spanish of the Airways Questionnaire 20, a short health related quality-of-life questionnaire for the clinical evaluation of asthma and chronic obstructive pulmonary disease]. AB - BACKGROUND AND OBJECTIVES: The routine use of health-related quality-of-life questionnaires in patients with chronic respiratory disease is limited due to the time required to complete them. The Airways Questionnaire 20 (AQ20) contains 20 easy-to-answer questions, making it ideal for use in routine practice. However, a Spanish version is not available. Our aim was to create a version of the AQ20 for use in Spain that would be equivalent to the original English questionnaire. METHOD: The original questionnaire was adapted using the translation backtranslation method. We evaluated the conceptual equivalence of the translation to the original questionnaire, classifying items as type A if they were fully equivalent, type B if they contained questionable wording, and type C if there were doubts about their equivalence. The items in categories B and C were re-examined by the researchers and translators, with input from the author of the original questionnaire and a group of patients. The final version was piloted among 30 patients. RESULTS: Low difficulty ratings were given to both the translation (3.45) and backtranslation (3.52). Sixty-five percent of the backtranslated items were considered to be fully equivalent to the original English items (type A), 15% were classified as type B, and 20% as type C. The final Spanish version, produced after discussion of items B and C, was administered to 15 patients with asthma (66% women; mean [SD] age, 53.13 [19.6] y) and 15 patients with chronic obstructive pulmonary disease (13% women; mean age, 67.8 [8.94] y). A Cronbach alpha of 0.92 was obtained. CONCLUSION: The adaptation process produced a Spanish questionnaire that is conceptually equivalent to the English original, easy to understand, and satisfactory in terms of internal consistency. PMID- 19186296 TI - [Modifications of diaphragmatic activity induced by midline laparotomy and changes in abdominal wall compliance]. AB - INTRODUCTION AND OBJECTIVE: Diaphragmatic activity varies with the initial length of the muscle. Our objective was to evaluate the influence of surgery and changes in abdominal wall compliance on diaphragmatic activity. METHODS: Both phrenic nerves in 7 mongrel dogs were stimulated electrically with single supramaximal pulses (twitch). The gastric (Pga) and transdiaphragmatic (Pdi) pressures generated and muscle shortening (sonomicrometry) were used to evaluate diaphragmatic activity, which was determined at baseline, after midline laparotomy, with an elastic abdominal bandage, and with a rigid circular cast. Abdominal pressure was then gradually increased in order to induce progressive lengthening of the diaphragm. RESULTS: After laparotomy, the pressures were somewhat lower (by 12%) than at baseline. The elastic bandage produced a slight increase in the pressure generated by the diaphragm (mean [SE] values: Pga, from 4.2 [0.3]cm H(2)O to 6.3 [0.9]cm H(2)O, P<.01; Pdi(tw), from 12.1 [2.0]cm H(2)O to 15.4 [1.8]cm H(2)O, P<.05]), and these values increased even further with the rigid cast (Pga, to 12.6 [1.5]cm H(2)O; Pdi, to 20.2 [2.3]cm H(2)O; P<.01 for both comparisons); this occurred despite smaller degrees of muscle shortening: by 57% [5%] of the initial length at functional residual capacity at baseline, by 49% [5%] with the bandage (P<.05), and by 39% [6%] with the cast (P<.01). With progressive lengthening of the muscle, its contractile efficacy increased up to a certain point (105% of the length at functional residual capacity), after which it began to decline. CONCLUSIONS: Abdominal wall compliance plays an important role in the diaphragmatic response to stimulation. This appears to be due mainly to changes in its length at rest. PMID- 19186297 TI - [Combination therapy for pulmonary arterial hypertension]. PMID- 19186298 TI - [The EPI-SCAN survey to assess the prevalence of chronic obstructive pulmonary disease in Spanish 40-to-80-year-olds: protocol summary]. AB - BACKGROUND AND OBJECTIVES: Chronic obstructive pulmonary disease (COPD) causes considerable morbidity and mortality in Spain. The 1997 IBERPOC study, applying the old criteria of the European Respiratory Society, reported a COPD prevalence of 9.1% in the adult population of Spain. The Epidemiologic Study of COPD in Spain (EPI-SCAN) aims to determine the current prevalence of COPD in residents of Spain aged 40-80 years and to estimate changes over the past 10 years. Secondary objectives are, among others, to describe the current prevalence of smoking and changes in COPD prevalence relative to previous studies; to describe treatments received by patients, quality of life, and the BODE index (body mass index, obstruction of airflow, dyspnea, and exercise tolerance); and to measure inflammatory markers in blood and exhaled-breath condensate. PATIENTS AND METHODS: EPI-SCAN is a population-based, cross-sectional epidemiologic study targeting the general population of Spain aged between 40 and 80 years. Participating centers were located in Barcelona, Burgos, Cordoba, Huesca, Madrid, Oviedo, Seville, Valencia, Vic, and Vigo. All subjects filled in an extensive questionnaire to collect social, demographic, and clinical information. Slow and forced spirometry tests before and after a bronchodilator test were also undertaken. Additionally, selected subjects performed a 6-minute walk test and answered generic and specific quality-of-life questionnaires, as well as an activities-of-daily-living questionnaire. Exhaled-breath condensate and blood samples were also collected from these subjects for measurement of inflammatory and other biomarkers. PMID- 19186299 TI - [Guidelines on asthma in extreme environmental conditions]. AB - Asthma is a highly prevalent chronic disease which, if not properly controlled, can limit the patient's activities and lifestyle. In recent decades, owing to the diffusion of educational materials, the application of clinical guidelines and, most importantly, the availability of effective pharmacological treatment, most patients with asthma are now able to lead normal lives. Significant social changes have also taken place during the same period, including more widespread pursuit of sporting activities and tourism. As a result of these changes, individuals with asthma can now participate in certain activities that were inconceivable for these patients only a few years ago, including winter sports, underwater activities, air flight, and travel to remote places with unusual environmental conditions (deserts, high mountain environments, and tropical regions). In spite of the publication of several studies on this subject, our understanding of the effects of these situations on patients with asthma is still limited. The Spanish Society of Pulmonology and Thoracic Surgery (SEPAR) has decided to publish these recommendations based on the available evidence and expert opinion in order to provide information on this topic to both doctors and patients and to avert potentially dangerous situations that could endanger the lives of these patients. PMID- 19186300 TI - [Diffuse interstitial lung disease related to peribronchiolar metaplasia]. AB - Peribronchiolar metaplasia is a histologic lesion characterized by fibrosis and bronchiolar epithelial proliferation, affecting peribronchiolar alveolar septa and terminal bronchioles. It has been considered a nonspecific tissue reaction secondary to the action of external factors, such as tobacco smoke and microbes, and is a common histologic finding in several diffuse interstitial lung diseases. Several such cases with clinical, radiologic, and lung function manifestations characteristic of idiopathic interstitial pneumonia have been described recently, all having peribronchiolar metaplasia as the only histologic abnormality. We report 2 cases of interstitial lung disease in which peribronchiolar metaplasia was the only finding of pathology. PMID- 19186301 TI - [Clown nose as an initial manifestation of squamous-cell lung carcinoma]. PMID- 19186302 TI - [Continous interscalene brachial plexus block: A rare cause of pleural effusion]. PMID- 19186303 TI - [Pleural effusion caused by pergolide]. PMID- 19186304 TI - Sinus inflammation. A draining problem. PMID- 19186305 TI - Low serotonin levels and seasonal affective disorder. PMID- 19186306 TI - Beyond the human genome. Meet the epigenome. PMID- 19186307 TI - My understanding is that calcium absorption relies on vitamin D and that vitamin D needs fat to be used by the body. If I drink low-fat milk with a low-fat meal, is the milk's supply of calcium wasted? PMID- 19186308 TI - I developed urinary stress incontinence after therapy for prostate cancer. I've tried regaining bladder control with conservative approaches. I also had a bulking agent injected into my urethra. It's been a year without much success. Are there any other options? PMID- 19186309 TI - Insulin therapy in very-low-birth-weight infants. PMID- 19186310 TI - Uric acid and cardiovascular risk. PMID- 19186311 TI - Uric acid and cardiovascular risk. PMID- 19186312 TI - Uric acid and cardiovascular risk. PMID- 19186313 TI - Uric acid and cardiovascular risk. PMID- 19186314 TI - Uric acid and cardiovascular risk. PMID- 19186315 TI - Self-esteem, confidence, and relationships in Brazilian men with erectile dysfunction receiving sildenafil citrate: a randomized, parallel-group, double blind, placebo-controlled study in Brazil. AB - INTRODUCTION: Psychosocial manifestations of erectile dysfunction (ED) differ across cultures. Understanding the treatment response to ED medications within cultural groups can aid in resource allocation and in developing treatment strategies. AIM: Evaluate the effect of sildenafil treatment on self-esteem, confidence, and sexual relationship satisfaction in Brazilian men with ED. MAIN OUTCOME MEASURE: The Self-Esteem and Relationship (SEAR) questionnaire, a validated, 14-question instrument developed to specifically address self-esteem and relationship issues within the context of ED. METHODS: Men aged 18 years or older with a clinical diagnosis of ED (< or = 21 on the Sexual Health Inventory for Men) and in a stable relationship with a partner during the study were eligible. The primary end point was a change from baseline in the self-esteem subscale of the SEAR questionnaire. Thirteen Brazilian sites participated in a randomized, double-blind, placebo-controlled trial of sildenafil treatment for ED. Patients were randomized to receive either 50 mg of sildenafil (adjustable to 25 mg or 100 mg based on patient response) or matching placebo approximately 1 hour before anticipated sexual activity but not more than once a day. RESULTS: At the end of double-blind treatment, 63 and 66 patients in the placebo and sildenafil groups, respectively, from 13 Brazilian sites were assessed for efficacy. Brazilian patients receiving sildenafil had significantly greater improvements in their scores on the SEAR self-esteem subscale (42.9 [95% confidence interval 35.7-50.0]) compared with placebo (21.1 [95% confidence interval 13.7-28.6]; P < 0.0001). Effect sizes ranged from 0.91 to 1.25 for individual SEAR components. CONCLUSIONS: The psychosocial parameters in Brazilian men with ED assessed by the SEAR questionnaire showed significant improvements in self-esteem, confidence, and relationships after treatment with sildenafil. PMID- 19186316 TI - [Evaluation of a novel seldinger-needle for computed tomography guided interventions: initial experiences]. AB - PURPOSE: To evaluate a new Seldinger puncture device for computed tomography guided interventions under difficult conditions, to analyze applicability, and to investigate assets and drawbacks. MATERIALS AND METHODS: From November 2007 to March 2008, we performed CT-guided interventions in 16 patients (7 women, 9 men; mean age 62 years old) using a new 20G-Seldinger needle (Sika-Med, Wiehl, Germany). This novel needle serves as a guide for many different interventional devices due to a guide wire welded on the proximal needle end. It allows continuous application of anesthesia via four tiny holes at the distal needle end until the region of interest is reached. Each intervention was subject to difficult interventional conditions. The indications for intervention were drainage (n = 7), Trucut biopsy of tumor (n = 8) and radiofrequency ablation (n = 1). Handling, success, advantages, drawbacks, complications and patient tolerance were noted after each procedure. A pain scale from 1 - 10 was used to grade the pain level during the intervention. RESULTS: All interventions were performed successfully and no severe complications were observed. Patient tolerance was very good resulting in a mean pain score of 2 +/- 1. Regions with dangerous and difficult access were successfully reached with the new Seldinger needle in 15 of 16 cases by dilatation of the puncture tract and continuous administration of local anesthesia via the system. Furthermore, different devices such as Trucut systems and a drainage catheter were able to be inserted without complication via the needle. With a proximal removable luer-lock connection, liquid material was able to be aspirated in six cases. CONCLUSION: Under difficult interventional conditions, the use of a Seldinger needle as a reliable technique for CT-guided interventions can provide a safe and successful procedure. PMID- 19186317 TI - [Endovascular therapy in a spontaneously appearing aneurysm in the left subclavian artery]. PMID- 19186318 TI - [Debate contribution, spinal surgery and economic conflict of interest]. PMID- 19186320 TI - [The Danish language and non-Danish physicians]. PMID- 19186319 TI - [Coronary artery abnormalities -- relevant or not?]. PMID- 19186321 TI - Retraction. Up-regulation of NOB1 in esophageal squamous cell carcinoma (p). PMID- 19186323 TI - Diverticulosis and diverticulitis. PMID- 19186324 TI - Subcutaneous abdominal adipose tissue subcompartments: potential role in rosiglitazone effects. AB - Abdominal visceral tissue (VAT) and subcutaneous adipose tissue (SAT), comprised of superficial-SAT (sSAT) and deep-SAT (dSAT), are metabolically distinct. The antidiabetic agents thiazolidinediones (TZDs), in addition to their insulin sensitizing effects, redistribute SAT suggesting that TZD action involves adipose tissue depot-specific regulation. We investigated the expression of proteins key to adipocyte metabolism on differentiated first passage (P1) preadipocytes treated with rosiglitazone, to establish a role for the diverse depots of abdominal adipose tissue in the insulin-sensitizing effects of TZDs. Adipocytes and preadipocytes were isolated from sSAT, dSAT, and VAT samples obtained from eight normal subjects. Preadipocytes (P1) left untreated (U) or treated with a classic differentiation cocktail (DI) including rosiglitazone (DIR) for 9 days were evaluated for strata-specific differences in differentiation including peroxisome proliferator-activated receptor-gamma (PPAR-gamma) and lipoprotein lipase (LPL) expression, insulin sensitivity via adiponectin and glucose transport-4 (GLUT4), glucocorticoid metabolism with 11 beta-hydroxysteroid dehydrogenase type-1 (11 beta HSD1), and alterations in the adipokine leptin. While depot-specific differences were absent with the classic differentiation cocktail, with rosiglitazone sSAT had the most potent response followed by dSAT, whereas VAT was resistant to differentiation. With rosiglitazone, universal strata effects were observed for PPAR-gamma, LPL, and leptin, with VAT in all cases expressing significantly lower basal expression levels. Clear dSAT-specific changes were observed with decreased intracellular GLUT4. Specific sSAT alterations included decreased 11 beta HSD1 whereas secreted adiponectin was potently upregulated in sSAT with respect to dSAT and VAT. Overall, the subcompartments of SAT, sSAT, and dSAT, appear to participate in the metabolic changes that arise with rosiglitazone administration. PMID- 19186325 TI - Trypanosoma cruzi infection of cultured adipocytes results in an inflammatory phenotype. AB - Infection with Trypanosoma cruzi, the etiologic agent of Chagas disease is accompanied by an intense inflammatory reaction. Our laboratory group has identified adipose tissue as one of the major sites of inflammation during disease progression. Because adipose tissue is composed of many cell types, we were interested in investigating whether the adipocyte per se was a source of inflammatory mediators in this infection. Cultured adipocytes were infected with the Tulahuen strain of T. cruzi for 48-96 h. Immunoblot and quantitative PCR (qPCR) analyses demonstrated an increase in the expression of proinflammatory cytokines and chemokines, including interleukin (IL)-1 beta, interferon-gamma, tumor necrosis factor-alpha, CCL2, CCL5, and CXCL10 as well as an increase in the expression of Toll-like receptors-2 and 9 and activation of the notch pathway. Interestingly, caveolin-1 expression was reduced while cyclin D1 and extracellular signal-regulated kinase (ERK) expression was increased. The expression of PI3kinase and the activation of AKT (phosphorylated AKT) were increased suggesting that infection may induce components of the insulin/IGF-1 receptor cascade. There was an infection-associated decrease in adiponectin and peroxisome proliferator-activated receptor-gamma (PPAR-gamma). These data provide a mechanism for the increase in the inflammatory phenotype that occurs in T. cruzi-infected adipocytes. Overall, these data implicate the adipocyte as an important target of T. cruzi, and one which contributes significantly to the inflammatory response observed in Chagas disease. PMID- 19186326 TI - Bingeing, self-restriction, and increased body weight in rats with limited access to a sweet-fat diet. AB - OBJECTIVE: Prior research has shown that fasting alternated with a diet of standard rodent chow and a 10% sucrose solution produces bingeing on the sucrose, but animals remain at normal body weight. The present study investigated whether restricted access to a highly palatable combination of sugar and fat, without food deprivation, would instigate binge eating and also increase body weight. METHODS AND PROCEDURES: Male rats were maintained for 25 days on one of four diets: (i) sweet-fat chow for 2 h/day followed by ad libitum standard chow, (ii) 2-h sweet-fat chow only 3 days/week and access to standard chow the rest of the time, (iii) ad libitum sweet-fat chow, or (iv) ad libitum standard chow. RESULTS: Both groups with 2-h access to the sweet-fat chow exhibited bingeing behavior, as defined by excessively large meals. The body weight of these animals increased due to large meals and then decreased between binges as a result of self restricted intake of standard chow following binges. However, despite these fluctuations in body weight, the group with 2-h access to sweet-fat chow every day gained significantly more weight than the control group with standard chow available ad libitum. DISCUSSION: These findings may have implications for the body weight fluctuations associated with binge-eating disorder, as well as the relationship between binge eating and the obesity epidemic. PMID- 19186328 TI - Baseline correlates of insulin resistance in inner city high-BMI African-American children. AB - To characterize the influence of diet-, physical activity-, and self-esteem related factors on insulin resistance in 8- 10-year-old African-American (AA) children with BMI greater than the 85th percentile who were screened to participate in a community-based type 2 diabetes mellitus (T2DM) prevention trial. In 165 subjects, fasting glucose- and insulin-derived values for homeostasis model assessment of insulin resistance (HOMA-IR) assessed insulin resistance. Body fatness was calculated following bioelectrical impedance analysis, and fitness was measured using laps from a 20-m shuttle run. Child questionnaires assessed physical activity, dietary habits, and self-esteem. Pubertal staging was assessed using serum levels of sex hormones. Parent questionnaires assessed family demographics, family health, and family food and physical activity habits. Girls had significantly higher percent body fat but similar anthropometric measures compared with boys, whereas boys spent more time in high-intensity activities than girls. Scores for self-perceived behavior were higher for girls than for boys; and girls desired a more slender body. Girls had significantly higher insulin resistance (HOMA-IR), compared with boys (P < 0.01). Adjusting for age, sex, pubertal stage, socioeconomic index (SE index), and family history of diabetes, multivariate regression analysis showed that children with higher waist circumference (WC) (P < 0.001) and lower Harter's scholastic competence (SC) scale (P = 0.044) had higher insulin resistance. WC and selected self-esteem parameters predicted insulin resistance in high-BMI AA children. The risk of T2DM may be reduced in these children by targeting these factors. PMID- 19186327 TI - Review and meta-analysis of pharmacotherapy for binge-eating disorder. AB - This study evaluated available controlled treatment studies to determine utility of pharmacotherapy for binge-eating disorder (BED). The authors identified randomized placebo-controlled trials testing pharmacotherapy-only treatments and controlled trials testing pharmacotherapy with psychotherapy treatments. Meta analysis was performed on placebo-controlled trials with data for attrition, remission, and weight loss. Qualitative review was performed on remaining controlled treatment literature. A total of 33 studies were considered of which 14 studies with a total of 1,279 patients were included in the meta-analysis of pharmacotherapy-only treatment and 8 studies with a total of 683 patients were included in the qualitative review of pharmacotherapy combined with psychotherapy interventions. No evidence suggested significant differences between medication and placebo for attrition. Evidence suggested that pharmacological treatments have a clinically significant advantage over placebo for achieving short-term remission from binge eating (48.7% vs. 28.5%) and for weight loss, although weight losses are not substantial. No data exist to allow evaluation of longer term effects of pharmacotherapy-only treatment for BED. Combining medications with psychotherapy interventions failed to significantly enhance binge outcomes, although specific medications (orlistat, topiramate) enhanced weight losses achieved with cognitive behavioral therapy and behavioral weight loss. In summary, BED patients can be advised that certain pharmacotherapies may enhance likelihood of stopping binge eating short term, but that longer-term effects are unknown. Although some weight loss may occur, it is unlikely to be substantial with available medications. Combining medications with cognitive or behavioral treatments is unlikely to enhance binge outcomes, but specific medications (orlistat, topiramate) may enhance weight losses, albeit modestly. PMID- 19186329 TI - Postprandial cytokine concentrations and meal composition in obese and lean women. AB - The aim of this study was to compare the acute effect of (i) meals rich in saturated fat, oleic acid, and alpha-linolenic acid and (ii) meals rich in starch and fiber on markers of inflammation and oxidative stress in obese and lean women. In a crossover study, 15 abdominally obese women (age, 54 +/- 9 years; BMI, 37.3 +/- 5.5 kg/m2) and 14 lean women (age, 53 +/- 10 years; BMI, 22.9 +/- 1.9 kg/m2) consumed meals rich in cream (CR), olive oil (OL), canola oil (CAN), potato (POT), and All-Bran (BRAN) in random order. Blood samples were collected before and up to 6 h after the meals and plasma interleukin-6 (IL-6), IL-8, tumor necrosis factor-alpha (TNF-alpha), lipid peroxides (LPOs), free-fatty acids (FFAs), insulin, glucose, and cortisol were measured. Plasma IL-6 decreased significantly 1 h after the meals then increased significantly above baseline at 4h and 6h in obese women and at 6h in lean women. The incremental area under the curve (iAUC) for IL-6 was significantly (P = 0.02) higher in obese compared with lean women and was significantly lower following the high fiber BRAN meal compared with a POT meal (P = 0.003). Waist circumference (R = 0.491, P = 0.007) and cortisol AUC (R = -0.415, P = 0.03) were significant determinants of the magnitude of 6h changes in plasma IL-6 after the meals. These findings suggest that the postprandial response of plasma IL-6 concentrations may be influenced by the type of carbohydrate in the meal, central adiposity, and circulating cortisol concentrations in women. PMID- 19186330 TI - Beta-aminoisobutyric acid prevents diet-induced obesity in mice with partial leptin deficiency. AB - Beta-Aminoisobutyric acid (BAIBA), a thymine catabolite, increases fatty acid oxidation (FAO) in liver and reduces the gain of body fat mass in Swiss (lean) mice fed a standard chow. We determined whether BAIBA could prevent obesity and related metabolic disorders in different murine models. To this end, BAIBA (100 or 500 mg/kg/day) was administered for 4 months in mice totally deficient in leptin (ob/ob). BAIBA (100 mg/kg/day) was also given for 4 months in wild-type (+/+) mice and mice partially deficient in leptin (ob/+) fed a high-calorie (HC) diet. BAIBA did not limit obesity and hepatic steatosis in ob/ob mice, but reduced liver cytolysis and inflammation. In ob/+ mice fed the HC diet, BAIBA fully prevented, or limited, the gain of body fat, steatosis and necroinflammation, glucose intolerance, and hypertriglyceridemia. Plasma beta hydroxybutyrate was increased, whereas expression of carnitine palmitoyltransferase-1 was augmented in liver and white adipose tissue. Acetyl CoA carboxylase was more phosphorylated, and de novo lipogenesis was less induced in liver. These favorable effects of BAIBA in ob/+ mice were associated with a restoration of plasma leptin levels. The reduction of body adiposity afforded by BAIBA was less marked in +/+ mice. Finally, BAIBA significantly stimulated the secretion of leptin in isolated ob/+ adipose cells, but not in +/+ cells. Thus, BAIBA could limit triglyceride accretion in tissues through a leptin-dependent stimulation of FAO. As partial leptin deficiency is not uncommon in the general population, supplementation with BAIBA may help to prevent diet-induced obesity and related metabolic disorders in low leptin secretors. PMID- 19186331 TI - Effects of pravastatin on obesity, diabetes, and adiponectin in diet-induced obese mice. AB - OBJECTIVE: The aim of this study was to investigate the in vivo effects of pravastatin on the development of obesity and diabetes in diet-induced obese (DIO) mice. METHODS AND PROCEDURES: We examined food intake, body-weight changes, visceral white adipose tissue (WAT) adiponectin and resistin levels, and energy metabolism. RESULTS: Treatment with 100 mg/kg/day pravastatin for 28 days decreased diet-induced weight gain and visceral adiposity. In addition, the weight of the WAT, the triglyceride (TG) contents of the liver and muscles, and the levels of serum insulin improved in the pravastatin-treated DIO mice. Furthermore, pravastatin treatment changed the WAT adiponectin and resistin mRNA expression and serum levels compared with the controls. Finally, pravastatin treatment increased oxygen consumption and decreased the respiratory quotient (RQ). DISCUSSION: Pravastatin treatment prevents the development of obesity and diabetes in DIO mice. The prevention of obesity may be mediated by increased oxygen consumption and a decrease in the RQ. These results provide novel insights into the use of pravastatin as a therapeutic tool for metabolic syndromes. PMID- 19186332 TI - A pilot of a video game (DDR) to promote physical activity and decrease sedentary screen time. AB - OBJECTIVE: We examined the feasibility of Dance Dance Revolution (DDR), a dance video game, in participants' homes, to increase physical activity (PA) and to decrease sedentary screen time (SST). METHODS AND PROCEDURES: Sixty children (7.5 +/- 0.5 years) were randomized in a 2:1 ratio to DDR or to wait-list control (10 week delay). DDR use was logged, PA was measured objectively by accelerometry. SST was self-reported at weeks 0 and 10. At week 28, after both groups had access to DDR, accelerometry and SST were repeated. RESULTS: Mean use of DDR was 89 +/- 82 (range 0-660 min) min per week (mpw). The DDR group showed increased vigorous PA and a reduction in light PA; the control group showed no increase in moderate and/or vigorous PA (MVPA) although they also had a reduction in light PA. Differences between the groups were not observed. The DDR group also reported a decrease in SST of -1.2 +/- 3.7 h per week (hpw) (P < 0.05), whereas the controls reported an increase of +3.0 +/- 7.7 hpw (nonsignificant). The difference in SST between the groups was significant, with less SST in the DDR group. Between weeks 10 and 28, numeric reductions in SST were reported in both groups. In the DDR group, SST at week 28 (8.8 +/- 6.0 hpw) was lower than baseline (10.5 +/- 5.5 hpw; P < 0.03). DISCUSSION: This pilot study suggests that DDR reduces SST and may facilitate slight increases in vigorous PA. Further study is needed to better characterize children and contexts in which DDR may promote a healthy lifestyle. PMID- 19186333 TI - Association of codon 16 and codon 27 beta 2-adrenergic receptor gene polymorphisms with obesity: a meta-analysis. AB - OBJECTIVE: To search for an association between the Glu27Gln (rs1042714; B27) and the Arg16Gly (rs1042713; B16) polymorphisms of the beta 2-adrenergic receptor (ADRB2) gene and obesity. METHODS: Meta-analysis of published studies, included if subjects were genotyped at either codon 27 ("B27") or codon 16 ("B16") of the ADRB2 gene and both obese and nonobese subjects were selected, based on a reported cutoff BMI limit. Initial selection included 14,444 subjects genotyped at B27 (rs1042714) and 6,825 genotyped at B16 (rs1042713). After testing each control group for Hardy-Weinberg equilibrium, the final selection included 10,404 subjects and 4,328 subjects, respectively. Studies were published before 18 August 2006. RESULTS: The frequency of Glu27 allele carriers, either homozygous or heterozygous, ranged from 6.71% in Aymara American Indians to 78.29% in a Dutch population. The frequency of Arg16 allele carriers varied from 51.4 to 64.6% in Europeans and from 71.1 to 85.6% in East Asians. The summary odds ratio (OR) from overall analyses showed no association between either rs1042714 or rs1042713 and obesity. In race groups with low Glu27 allele frequency (Asians, Pacific Islanders, and American Indians), ORs revealed a significant obesity risk associated with rs1042714. These results were not found in East Asians for rs1042713. DISCUSSION: The presence of the Glu27 allele in the ADRB2 gene appears to be a significant risk factor for obesity in Asians, Pacific Islanders, and American Indians, but not in Europeans. Obesity does not appear to be associated with the Arg16 allele. PMID- 19186334 TI - The incidence rate of seizures in relation to BMI in UK adults. AB - A retrospective cohort study using the data from The Health Improvement Network (THIN) database in the United Kingdom was conducted to examine the incidence rates of seizures across different BMI levels in the adult population aged > or = 18 years. Poisson regression was used to examine the relationship between BMI and seizures. The overall incidence rate of seizures was found to be 31.2 cases per 100,000 person-years. The incidence rate of seizures (cases per 100,000 person years) in obese patients (BMI > or = 30 kg/m2) was 34.8 (95% confidence interval (CI), 23.1, 46.4), comparable to that in patients with normal weight (BMI between 18.5 and 24.9 kg/m2) (35.8, 95% CI (26.6, 44.9)). In contrast, underweight patients (< 18.5 kg/m2) or extremely obese (> or = 40 kg/m2) patients tended to have higher incidence rates than those with normal weight. After adjustment for age, gender, and smoking status, compared to patients with normal weight, those who were underweight or extremely obese had a rate ratio (RR) for seizures of 1.6 (95% CI (0.7, 3.8)) and 1.7 (95% CI (0.7, 3.9)), respectively. To date, we have not found any study that examines the associations between BMI or obesity and seizures. In this study, the incidence rates of seizures in the extremely obese and underweight patients tended to be higher than that in the normal-weight patients. PMID- 19186335 TI - Energy expenditure and adiposity in Nigerian and African-American women. AB - OBJECTIVE: Obesity is a prevalent condition in industrialized societies and is increasing around the world. We sought to assess the relative importance of resting energy expenditure (REE) and activity EE (AEE) in two populations with different rates of obesity. METHODS AND PROCEDURES: Women of African descent between 18 and 59 years of age were recruited from rural Nigeria and from metropolitan Chicago. Total EE (TEE) was measured using the doubly labeled water (DLW) technique and REE by indirect calorimetry; AEE was calculated as the difference between TEE and the sum of REE plus a factor for the thermic effect of food. In the analyses all EE parameters were adjusted for body size using a regression method. Comparisons were made between the groups and associations between EE and adiposity examined. RESULTS: A total of 149 Nigerian and 172 African-American women completed the protocol. All body size measurements were lower in the Nigerian women. Adjusted TEE and REE were higher in the Nigerian cohort but adjusted AEE did not differ significantly. Adjustment for parity, seasonality, and recent illness did not modify mean AEE or adiposity. In neither cohort was there a meaningful association between measures of AEE and adiposity. DISCUSSION: In these cohorts of women from very different environments, AEE did not differ significantly nor was it associated cross-sectionally with adiposity. If generalizable, these findings suggest that reduction in AEE may have less of a role in the development of obesity than anticipated. The possibility remains that variation in type and duration of activity plays a role not captured by total AEE. PMID- 19186336 TI - The effect of obesity and cardiometabolic risk factors on expenditures and productivity in the United States. AB - OBJECTIVE: To examine the effect of obesity and cardiometabolic risk factors on medical expenditures and missed work days. METHODS AND PROCEDURES: The 2000 and 2002 Medical Expenditure Panel Survey (MEPS), a nationally representative survey of the US population, was used to estimate the marginal effect of obesity (BMI > or = 30) on annual per-person medical expenditures and missed work days for patients with diabetes, dyslipidemia, or hypertension using multivariate regression methods controlling for age, sex, race, ethnicity, education, income, insurance, and smoking status. Maximum Likelihood Heckman Selection with Smearing retransformation was used to assess medical expenditures, and Negative Binomial regression was used for missed work days. RESULTS: Normal weight individuals with diabetes, dyslipidemia, or hypertension had significantly greater medical expenditures than those without the respective condition ($6,006 (5,124-6,887), $4,760 (4,102-5,417), $3,911 (3,345-4,476)) and obesity significantly exacerbated this effect ($7,986 (7,397-8,574), $7,636 (7,072-8,200), $6,197 (5,745-6,649); $2007; all P < 0.05). In addition, diabetes, dyslipidemia, and hypertension resulted in greater missed work days (3.1 (0.94-6.21), 3.2 (0.42-7.91), 1.4 (0.0 3.52)) (all P < 0.05 except hypertension), which resulted in greater lost productivity ($433, $451, $199) and obesity significantly exacerbated the deleterious effect on work days (8.7 (4.44-15.2), 5.5 (2.18-10.5), 4.5 (2.92 6.34)) and lost productivity ($1,217, $763, $622) (all P < 0.05). In addition, medical expenditures increased for increasing weight category and increasing number of risk factors. DISCUSSION: Obesity significantly exacerbates the deleterious effect of diabetes, dyslipidemia, and hypertension on medical expenditures and productivity loss in the United States. Obesity is preventable and public health efforts need to be undertaken to prevent its alarming increase in order to reduce the incidence and effect of cardiometabolic risk factors. PMID- 19186338 TI - Reformulation of BMI and percent body fat to remove the height bias in 8-year olds. AB - BMI and percent body fat (%BF) are both related to height (Ht) in prepubertal children, so may misrepresent childhood adiposity, especially in tall or short children. We sought to construct replacement functions for BMI and %BF that are independent of Ht. Fat mass (FM) was measured using dual-energy X-ray absorptiometry, together with Ht and body mass (BM) in 746 healthy boys and girls aged 8 years (0.34 s.d.). Relationships between BM, FM, and Ht were measured and values of p and q derived such that the functions BM.Ht(-p) and FM.BM(-q) were unrelated to Ht. BM was not directly proportional to Ht2, BMI being significantly related to Ht in both boys and girls (P < 0.001). BM was proportional to Ht3, BM.Ht(-3) being independent of Ht. Similarly, FM was not directly proportional to BM and %BF was significantly related to Ht (P < 0.001). While FM was proportional to BM2, FM.BM(-1.5) was the function found to be independent of Ht. Using the 85th and 95th percentiles as the cutoffs for overweight and obesity respectively, 6.4% of the boys and 6.8% of the girls were classified differently by BMI and the Ht independent measure BM.Ht(-3). Similarly, 10.1% boys and 13.7% girls were classified differently by %BF and the Ht independent measure FM.BM(-1.5). We propose that improved diagnostic accuracy of body composition in 8-year-olds is provided by the BM function (BMF, BM.Ht(-3)) and FM function (FMF, FM.BM(-1.5)) replacing BMI and %BF, which both overestimate the adiposity of taller children and underestimate it in shorter children. PMID- 19186337 TI - Changes in body weight and waist circumference affect incident hypercholesterolemia during 7 years of follow-up. AB - OBJECTIVE: To assess whether changes in total and regional adiposity affect the odds for becoming hypercholesterolemic. METHODS AND PROCEDURES: Changes in BMI and waist circumference were compared to self-reported physician-diagnosed hypercholesterolemia in 24,397 men and 10,023 women followed prospectively in the National Runners' Health Study. RESULTS: Incident hypercholesterolemia were reported by 3,054 men and 519 women during (mean +/- s.d.) 7.8 +/- 1.8 and 7.5 +/ 2.0 years of follow-up, respectively. Despite being active, men's BMI increased by 1.15 +/- 1.71 kg/m2 and women's BMI increased by 0.96 +/- 1.89 kg/m2. The odds for developing hypercholesterolemia increased significantly in association with gains in BMI and waist circumferences in both sexes. A gain in BMI > or = 2.4 kg/m2 significantly (P < 0.0001) increased the odds for hypercholesterolemia by 94% in men and 129% in women compared to those whose BMI declined (40 and 76%, respectively, adjusted for average of the baseline and follow-up BMI, P < 0.0001). A gain of > or = 6 cm in waist circumference increased men's odds for hypercholesterolemia by 74% (P < 0.0001) and women's odds by 70% (P < 0.0001) relative to those whose circumference declined (odds increased 40% at P < 0.0001 and 49% at P < 0.01, respectively adjusted for average circumference). BMI and waist circumference at the end of follow-up were significantly associated (P < 0.0001) with the log odds for hypercholesterolemia in both men (e.g., coefficient +/- s.e.: 0.115 +/- 0.011 per kg/m2) and women (e.g., 0.119 +/- 0.019 per kg/m2) when adjusted for baseline values, whereas baseline BMI and circumferences were unrelated to the log odds when adjusted for follow-up values. DISCUSSION: These observations are consistent with the hypothesis that weight gain acutely increases the risk for hypercholesterolemia. PMID- 19186339 TI - Utilising and promoting public health and health services research in complementary and alternative medicine: the founding of NORPHCAM. PMID- 19186340 TI - Attitudes and beliefs towards complementary and alternative medicine (CAM): a cross-cultural approach comparing Singapore and the United Kingdom. AB - This study examined Eastern (Singapore) and Western (United Kingdom) attitudes and beliefs towards complementary and alternative medicine (CAM), and towards orthodox medicine (OM). In all 60 individuals from Singapore and 50 from the United Kingdom (total n = 110) filled in an 8-page questionnaire. UK participants were more willing to use various forms of CAM, and had a generally more optimistic view of the usefulness and efficacy of CAM. Singaporeans were more in favour of OM, prescription drugs, and general practitioners (GPs). Explanations for this observation include the differences in availability and accessibility of CAM in each country, as well as the medical models that are valued and supported in each culture. PMID- 19186341 TI - The prevalence and correlates of mind-body therapy practices in patients with acute coronary syndrome. AB - OBJECTIVES: While the benefits of mind-body therapy (MBT) for cardiac secondary prevention continues to be investigated, the prevalence of such practices by cardiac patients is not well known. The aim of this study was to quantitatively examine the prevalence of MBT practice and its sociodemographic, clinical, psychosocial and behavioral correlates among patients with acute coronary syndrome (ACS). METHODS: Six hundred and sixty-one ACS in-patients (75% response rate) recruited from three hospitals completed a demographic survey, and clinical data were extracted from charts. Four hundred and sixty five patients (81% retention rate; 110 (23.7%) female) responded to an 18-month post-discharge survey that queried about MBT use and its correlates. RESULTS: One hundred and sixty-three (35.1%) ACS patients practised MBT in their lifetime, and 118 (25.4%) were currently practising. MBT users were more often women (OR = 2.98), nonwhite (OR = 2.17), had higher levels of education (OR = 2.22), past smokers (OR = 3.33), reported poorer mental health (OR = 2.15), and engaged in more exercise (OR = 1.65). CONCLUSION: One-third of ACS patients practised some form of MBT. The greater MBT practice among female ACS patients is noteworthy, given their generally lower physical activity and lower receipt of evidence-based treatments including cardiac rehabilitation. In addition, there is some evidence that MBT can promote mental well-being, and thus such practice might reduce risk related to negative affect in cardiac patients. PMID- 19186342 TI - Topical application of Garcinia mangostana L. pericarp gel as an adjunct to periodontal treatment. AB - OBJECTIVE: To evaluate the effects of gel containing Garcinia mangostana L. pericarp extract (GM gel) applied topically as an adjunct to periodontal treatment. DESIGN: Subjects who had periodontal pockets on their single-rooted teeth were randomized into the test or control group. Subjects in the test group received periodontal treatment consisting of scaling, root planing and subgingival application of GM gel while those in the control group received scaling and root planing without GM gel application. SETTING: Mahidol University, Faculty of Dentistry, Thailand. MAIN OUTCOME MEASURES: Clinical parameters included probing pocket depth (PPD), clinical attachment level (CAL), bleeding on probing (BOP), Gingival Index (GI) and Plaque Index (PI). Microbiological parameter included subgingival microbial composition as examined by phase contrast microscopy. RESULTS: Clinical improvement compared to baseline was found in both groups (P < 0.05). The test group exhibited significantly higher reduction in mean PPD, GI and BOP than the control group at the 3rd month after treatment (P < 0.05). Subgingival microbial composition changed from diseased state to that compatible with health after treatment in both groups. However, significant differences between groups were found only in the mean percentage of cocci at the 1st and 3rd month after treatment (P < 0.05). CONCLUSIONS: GM gel could enhance the clinical effects of periodontal treatment. PMID- 19186343 TI - A comparison of effects on regional pressure pain threshold produced by deep needling of LI4 and LI11, individually and in combination. AB - OBJECTIVES: To compare the effects of unilateral and bilateral needling of the same acupoint, and the effects of individual and combined needling of two distinct acupoints on pressure pain threshold (PPT). DESIGN: 22 subjects completed the randomised, dual blind (subject and evaluator) repeated measures study. PPT was measured before and after each intervention at ten sites (acupoints and nonacupoints) across the body with an algometer. INTERVENTIONS: The same manual acupuncture techniques were applied to four interventions of large intestine 4 (LI4) unilaterally; LI4 bilaterally; large intestine 11 (LI 11) unilaterally; and LI4 in conjunction with LI11, both unilaterally. MAIN OUTCOME MEASURES: (1) Percentage change in PPT from preintervention baseline measured at the 10 regional sites following every intervention; (2) participants' perceptions of pain; needling sensations; tension during, and anxiety prior to, each intervention; and changes in practitioner behaviour. RESULTS: Following all four interventions, statistically significant increases in mean PPT were observed. These occurred at nine sites following the LI4 intervention either unilaterally or bilaterally; at six sites for LI11 intervention; and at five sites following the combined LI11 and LI4 intervention. These increases were significantly greater for the bilateral LI4 intervention than the unilateral LI4 intervention at only two sites (p < 0.02 and p < 0.0001). There were no statistically significant differences in the subjective perceptions among the four interventions. CONCLUSION: The enhanced effects on PPT by the bilateral compared with the unilateral intervention at LI4 although limited, do provide some support for the traditional Chinese medicine (TCM) assumption that bilateral needling of the same point enhances the treatment effect. There was no support for the assumption that combined needling of points from the same channel should enhance the treatment effect and failure to obtain better effects by combined needling of points from the same channel could result from the interaction occurring during the combined needling. PMID- 19186344 TI - Can individuals identify if needling was performed with an acupuncture needle or a non-penetrating sham needle? AB - A control treatment in acupuncture research must be credible, regardless if the needling is performed by one or by several therapists. OBJECTIVE: To investigate if individuals could identify whether needling had been given with an acupuncture needle or a sham needle and if the therapist influenced this ability. DESIGN: Eighty individuals were randomized to one single needling given by one of four physiotherapists using either an invasive needle or a non-penetrating telescopic sham needle. RESULTS: An equal proportion of individuals, 27 (68%), in the acupuncture group and the sham group answered incorrectly or was not sure at all regarding needling type but the proportion varied between the therapists from 55 to 80% (ns). Bang's blinding index was 0.20 (95% CI 0.03-0.36) in the acupuncture group and 0.10 (95% CI 0.09-0.29) in the sham group (interpretation: 20 and 10% identified needling type beyond statistical chance). Acupuncture was on a four grade scale rated as median "mildly painful" and sham as "not painful" (ns). Pain ratings varied from median "not" to "mildly painful" in the therapists (p = 0.01). CONCLUSIONS: Two thirds of individuals needled by acupuncture as well as sham could not identify needling type and only 10-20% of the individuals were unblinded beyond chance. The therapists, not the needling type, influenced how painful the needling was perceived. IMPLICATIONS: To achieve blinding success in acupuncture efficacy studies using the sham needle, the needling procedure must be strictly standardized in order to minimize differences between the therapists. PMID- 19186345 TI - Treatment regimens of acupuncture for low back pain--a systematic review. AB - OBJECTIVES: To investigate the difference in acupuncture treatment regimens for low back pain (LBP) among textbooks, clinical studies and clinical practice, and explore reasons for such differences. METHODS: A systematic review of textbooks, surveys, clinical studies (randomised controlled trials (RCTs) and case studies) published in English or Chinese, was conducted, plus Chinese expert opinions. Data on detailed acupuncture treatment regimens were extracted. RESULTS: 12 textbooks in English, 31 textbooks in Chinese, 3 surveys in English, no surveys in Chinese, 9 Chinese expert opinions, 25 RCTs in English, 29 RCTs in Chinese, and 38 case studies in English were included. For chronic non-specific LBP, the outstanding differences in regimens found were (irrespective of the approach of acupuncture): (i) clinical studies and surveys (9-11 points) reported the use of many more points per session than Chinese experts (5 points); (ii) two extreme frequencies of treatments were used across all sources (1-2 times/week vs. 5-6 times/week). For acute LBP, the notable differences between RCTs and case studies were: (i) a variety of special points were investigated; (ii) case studies reported the use of many more points per session (10 points) than RCTs in Chinese (2 points); (iii) RCTs in Chinese administrated more treatments (6.5 sessions) than RCTs in English (3.5 sessions) and case studies (3 sessions). CONCLUSION: For non-specific LBP, treatment regimens of acupuncture differ by the types of reference sources, in terms of treatment frequency, the points chosen, number of points needled per session, duration and sessions, and co-interventions. PMID- 19186346 TI - Bringing oral health care to school-aged children. PMID- 19186347 TI - Comparison of 5 flow cytometric immunophenotyping systems for absolute CD4+ T lymphocyte counts in HIV-1-infected patients living in resource-limited settings. AB - Enumeration of CD4+ T lymphocytes is important in management of HIV-infected patients. However, CD4 testing by current gold standard bead-based flow cytometer (FCM) system is expensive for developing countries. This study compared 2 affordable volumetric FCMs with the 3 predicate FCM systems. CD4+ T-lymphocyte counts on blood samples from 150 HIV-1-infected Thai patients were determined in parallel by 5 FCM systems: the 2 single-platform volumetric FCM systems, Guava and CyFlow(green); the 2 standard single-platform bead-based systems (2-color FACSCount and the TriTEST/TruCOUNT tube using a FACSCalibur FCM); and the dual platform TriTEST system. Correlation and agreement were analyzed using linear regression and Bland-Altman analysis. Results from these 2 volumetric systems gave similar results and excellent correlation: R2 > 0.93; mean biases ranged from +6.3 to +24.1 cells per microliter more for the Guava. In contrast, the CyFlow(green) showed the lowest values with R2 > 0.97; mean biases ranged from 9.8 to -27.6 cells per microliter. This indicates that the absolute CD4+ T lymphocyte counts determined by CyFlow(green) are < FACSCount < DP TriTEST < TriTEST/TruCOUNT < Guava. Although the use of these 2 volumetric FCMs could make CD4+ T-lymphocyte enumeration more affordable in resource-poor settings, variations among these systems should be considered if these are to be interchanged. PMID- 19186348 TI - Characterization of quantitative and functional innate immune parameters in HIV-1 infected Colombian children receiving stable highly active antiretroviral therapy. AB - BACKGROUND: The immunological benefits of highly active antiretroviral therapy (HAART) in HIV-1-infected children include reconstitution of CD4+ T-cell count and functional activity. The effect of HAART on innate immune cells has not been well established. AIM: To characterize innate immune responses in HAART-treated HIV-1-infected children. PATIENTS AND METHODS: 23 HIV-1-infected children on stable HAART and 23 uninfected children were evaluated. The frequency of innate immune cells in peripheral blood was determined by flow cytometry and functional activity was evaluated using Toll-like receptor agonists. RESULTS: Compared with uninfected children, HAART-treated HIV-1-infected children exhibited a significant decrease in the frequency of plasmacytoid dendritic cells and natural killer and T-cell receptor (TCR)-invariant CD1d-restricted T cells. This deficiency of innate immune cells was observed mainly in children with detectable viral load. We also compared the magnitude of the quantitative restoration of those cells comparing HIV-1-infected children with HIV-1-infected adults and found a partial effect of HAART on immune restoration that was independent of age. In both pediatric and adult subjects Toll-like receptor agonists induced expression of costimulatory molecules and production of proinflammatory cytokines by dendritic cells. Peripheral blood mononuclear cells of HIV-1-infected children produced significantly reduced amounts of interferon-alpha compared with uninfected children. CONCLUSIONS: HAART administration to HIV-1-infected children does not lead to a complete increase of circulating innate immune cells, particularly in patients with incomplete suppression of HIV. PMID- 19186349 TI - Ritonavir greatly impairs CYP3A activity in HIV infection with chronic viral hepatitis. AB - PURPOSE: Ritonavir is a powerful inhibitor of cytochrome P450 3A (CYP3A) that metabolizes many antiretrovirals. We examined the effect of ritonavir and of chronic viral hepatitis (CVH) status on CYP3A activity. METHODS: Twenty-six HIV positive men (13 with CVH, 16 on chronic ritonavir-based highly active antiretroviral therapy) received oral and intravenous midazolam, a probe for CYP3A phenotypic activity. RESULTS: CYP3A activity was expressed as oral clearance of the midazolam probe. In HIV-positive subjects not on ritonavir, CYP3A activity (mean +/- SD) did not differ between subjects by CVH (no CVH, controls: 28.5 +/- 9.0 vs. CVH+: 23.2 +/- 6.2 mL/min/kg, not significant). In those on ritonavir (R), CYP3A activity was 7% of controls (R: 2.1 +/- 0.8 vs. no R 28.5 +/- 9.0 mL/min/kg, P < 0.0004). CYP3A activity in subjects on ritonavir and with CVH was further reduced to 4% of controls (no CVH, R+ 2.1 +/- 0.8 vs. R+, CVH+ 1.0 +/- 0.4 mL/min/kg, P < 0.006). CONCLUSIONS: Ritonavir markedly decreases CYP3A activity. In the presence of CVH, ritonavir-based therapy further reduces CYP3A activity by half. Coinfection with CVH impairs CYP3A activity in the presence of the CYP3A inhibitor ritonavir. PMID- 19186350 TI - Antiretroviral therapy exposure and insulin resistance in the Women's Interagency HIV study. AB - BACKGROUND: Evidence suggesting an increased risk of cardiovascular disease in HIV-infected individuals has heightened the need to understand the relation of HIV infection, antiretroviral therapy use, and non-HIV-related factors with insulin resistance (IR). METHODS: Prospective study of 1614 HIV-infected and 604 HIV-uninfected participants from the Women's Interagency HIV Study between October 2000 and March 2007. Homeostasis model assessment (HOMA)-estimated IR at 11,019 semiannual visits. RESULTS: HIV-infected women reporting highly active antiretroviral therapy (HAART) had higher median HOMA than HIV-uninfected women {1.20 [95% confidence interval (CI): 1.11 to 1.30] times higher for those reporting protease inhibitor-containing HAART; 1.10 (95% CI: 1.01 to 1.20) times higher for those reporting non-protease inhibitor-containing HAART}. Among HIV infected, cumulative exposure to nucleoside reverse transcriptase inhibitors (NRTIs) of > 3 years was associated with HOMA 1.13 (95% CI: 1.02 to 1.25) times higher than the HOMA without any cumulative NRTI exposure. Cumulative exposure to the NRTI stavudine of > 1 year was associated with HOMA 1.15 (95% CI: 1.05 to 1.27) times higher than the HOMA without any cumulative stavudine use. Family history of diabetes, hepatitis C virus seropositivity, higher body mass index, or reporting menopause was associated with higher HOMA. CINCLUSIONS: Longer cumulative exposure to NRTI; in particular, stavudine is associated with greater IR in HIV-infected women. PMID- 19186351 TI - The International HIV Dementia Scale is a useful screening tool for HIV associated dementia/cognitive impairment in HIV-infected adults in Yaounde Cameroon. AB - OBJECTIVE: As a baseline for a series of studies on HIV-associated dementia (HAD), we sought to assess the usefulness of the recently developed International HIV Dementia Scale (IHDS) as a screening tool for HAD or HIV-associated cognitive impairment (HACI) in HIV-positive adults in Yaounde-Cameroon. DESIGN: The frequency of HAD/HACI is largely unknown in resource-limited countries. In Cameroon, few studies suggest that HAD may be frequent but no specific study had so far investigated the problem. We therefore used a case-control study design involving HIV-positive adults as cases and HIV-negative individuals as controls to determine the usefulness of the IHDS as a screening instrument. METHODS: HIV positive adults followed up in an HIV outpatient clinic were matched to HIV negative subjects for age and sex and screened using IHDS. RESULTS: Overall, 204 HIV-positive individuals and 204 HIV-negative subjects were screened. The HIV positive subjects had a significantly lower IHDS mean total score of 10.87 compared with the HIV-negative subjects with a score of 11.28 (P = 0.00). Abnormal scores (< or = 10) on the IHDS were found in 21.1% of the HIV-positive subjects and in 2.5% of the HIV-negative subjects (P = 5.0 x 10(-10)). CONCLUSIONS: These results suggest that the prevalence of possible HAD/HACI may be higher in Cameroon than the previous estimates and demonstrate that the IHDS can be used as a screening tool for HAD in Cameroon. We therefore suggest that all studies on HAD in Cameroon should strategically start with the IHDS as a screening tool. PMID- 19186352 TI - HIV seropositivity predicts longer duration of stay and rehospitalization among nonbacteremic febrile injection drug users with skin and soft tissue infections. AB - BACKGROUND: Skin/soft tissue infections (SSTIs) are the leading cause of hospital admissions among injection drug users (IDUs). METHODS: We performed a retrospective investigation to determine the epidemiology of SSTIs (ie, cellulitis and/or abscesses) in febrile IDUs, with a focus on bacteriology and potential predictors of increased health care utilization measured by longer length of stay and rehospitalization. Subjects were drawn from a cohort of febrile IDUs presenting to an inner-city emergency department from 1998 to 2004. RESULTS: Of the 295 febrile IDUs with SSTIs, specific discharge diagnoses were cellulitis only (n = 143, 48.5%), abscesses only (n = 113, 38.3%), and both (n = 39, 13.2%). Documented HIV infection rate was 28%. Of note, 10 subjects were newly diagnosed with HIV infection during their visits. Staphylococcus aureus was the leading pathogen, and increasing rates of methicillin-resistant S. aureus emerged over time (before 2001: 4%, 2001-2004: 56%, P < 0.01). HIV seropositivity predicted rehospitalization within 90 days [adjusted hazard ratios and 95% confidence intervals: 2.90 (1.20 to 7.02)]. HIV seropositivity also predicted increased length of stay in those who were nonbacteremic [adjusted hazard ratios and 95% confidence intervals: 1.49 (1.11 to 2.01)]. CONCLUSIONS: Among febrile IDUs with SSTIs, a strong association between HIV seropositivity and health care resource utilization was found. Accordingly, attention to HIV serostatus should be considered in clinical disposition decisions for this vulnerable high-risk population. PMID- 19186353 TI - Comparison of cervicovaginal lavage, cervicovaginal lavage enriched with cervical swab, and vaginal tampon for the detection of HIV-1 RNA and HSV-2 DNA in genital secretions. AB - METHODS: We compared the performance of 3 collection methods for cervicovaginal secretions [cervicovaginal lavage (CVL), CVL enriched with a cervical swab (eCVL), and vaginal tampon (VT)] to identify the most reliable method for detection of cervicovaginal HIV-1 and herpes simplex virus type 2 (HSV-2). HIV-1 RNA (Nuclisens EasyQ; BioMerieux, Marcy-l'Etoile, France), HSV-2 DNA (real-time polymerase chain reaction), and microscopic blood and semen traces were detected in samples from 19 HIV-1-HSV-2-coinfected women seen at 4 weekly visits. RESULTS: HIV-1 RNA was detected in 49 (79%) of 62 eCVLs, 41 (61%) of 67 CVLs, and 27 (57%) of 47 VTs. Detection of HIV-1 RNA was higher in eCVL compared with CVL [45/58 (78%) vs. 32/58 (55%); risk ratio 1.41, 95% confidence interval 1.05 to 1.88]. CONCLUSIONS: Although more eCVLs were contaminated with microscopic blood (29%) than CVLs (22%) or VTs (7%), detection of HIV-1 RNA remained higher using eCVL compared with CVL (risk ratio 1.43, 95% confidence interval 1.02 to 2.02) in uncontaminated samples. HSV-2 DNA was detected in less than 10% of samples by each method but in 7 (37%) of 19 women overall by 1 or more methods. PMID- 19186354 TI - Estimating the lost benefits of antiretroviral drug use in South Africa. AB - South Africa is one of the countries most severely affected by HIV/AIDS. At the peak of the epidemic, the government, going against consensus scientific opinion, argued that HIV was not the cause of AIDS and that antiretroviral (ARV) drugs were not useful for patients and declined to accept freely donated nevirapine and grants from the Global Fund. Using modeling, we compared the number of persons who received ARVs for treatment and prevention of mother-to-child HIV transmission between 2000 and 2005 with an alternative of what was reasonably feasible in the country during that period. More than 330,000 lives or approximately 2.2 million person-years were lost because a feasible and timely ARV treatment program was not implemented in South Africa. Thirty-five thousand babies were born with HIV resulting in 1.6 million person-years lost by not implementing a mother-to-child transmission prophylaxis program using nevirapine. The total lost benefits of ARVs are at least 3.8 million person-years for the period 2000-2005. PMID- 19186355 TI - Group sex events and HIV/STI risk in an urban network. AB - OBJECTIVES: To describe: (a) the prevalence and individual and network characteristics of group sex events (GSEs) and GSE attendees; and (b) HIV/sexually transmitted infection (STI) discordance among respondents who said they went to a GSE together. METHODS AND DESIGN: In a sociometric network study of risk partners (defined as sexual partners, persons with whom respondents attended a GSE, or drug injection partners) in Brooklyn, NY, we recruited a high risk sample of 465 adults. Respondents reported on GSE attendance, the characteristics of GSEs, and their own and others' behaviors at GSEs. Sera and urines were collected, and STI prevalence was assayed. RESULTS: Of the 465 participants, 36% had attended a GSE in the last year, 26% had sex during the most recent of these GSEs, and 13% had unprotected sex there. Certain subgroups (hard drug users, men who have sex with men, women who have sex with women, and sex workers) were more likely to attend and more likely to engage in risk behaviors at these events. Among 90 GSE dyads, in which at least 1 partner named the other as someone with whom they attended a GSE in the previous 3 months, STI/HIV discordance was common [herpes simplex virus (HSV-2): 45% of dyads, HIV: 12% of dyads, and chlamydia: 21% of dyads]. Many GSEs had 10 or more participants, and multiple partnerships at GSEs were common. High attendance rates at GSEs among members of large networks may increase community vulnerability to STI/HIV, particularly because network data show that almost all members ofa large sociometric risk network either had sex with a GSE attendee or had sex with someone who had sex with a GSE attended. CONCLUSIONS: Self-reported GSE attendance and participation were common among this high-risk sample. STI/HIV discordance among GSE attendees was high, highlighting the potential transmission risk associated with GSEs. Research on sexual behaviors should incorporate measures of GSE behaviors as standard research protocol. Interventions should be developed to reduce transmission at GSEs. PMID- 19186357 TI - Housing status and HIV risk behaviors among homeless and housed persons with HIV. AB - OBJECTIVES: To compare drug, alcohol, and sexual HIV transmission risk behaviors of homeless and housed people living with HIV/AIDS. METHODS: Data were from 8075 respondents in a cross-sectional, multisite behavioral survey of adults recently reported to have HIV infection. RESULTS: At interview, 310 respondents (4%) were homeless. Compared with homeless respondents, housed respondents were more likely to be sexually active (past 12 months). However, sexually active homeless respondents had more sex partners (lifetime and past 12 months), greater sex exchange for money or drugs (lifetime and past 12 months), and greater unprotected vaginal or anal sex with an unknown serostatus partner. Homeless respondents were more likely to have possible alcohol abuse (lifetime), used drugs (last 12 months), and injected drugs (lifetime and past 12 months). After controlling for potential confounding variables, housing status remained a significant predictor of number of sex partners (past 12 months), sex exchange (lifetime and past 12 months), unprotected sex with unknown status partner, and all drug and alcohol use variables. CONCLUSIONS: Homeless people living with HIV/AIDS are more likely to have ever or recently engaged in substance use and HIV transmission risk behaviors. Findings underscore the need to provide HIV prevention services to homeless persons and address their housing needs. PMID- 19186356 TI - Predictors of sharing injection equipment by HIV-seropositive injection drug users. AB - Among HIV-positive injection drug users (IDUs), we examined baseline predictors of lending needles and syringes and sharing cookers, cotton, and rinse water in the prior 3 months at follow-up. Participants were enrolled in Intervention for Seropositive Injectors-Research and Evaluation, a secondary prevention intervention for sexually active HIV-positive IDUs in 4 US cities during 2001 2005. The analyses involved 357 participants who reported injecting drugs in the prior 6 months at either the 6- or 12-month follow-up visit. About half (49%) reported at least 1 sharing episode. In adjusted analyses, peer norms supporting safer injection practices and having primary HIV medical care visits in the prior 6 months were associated with reporting no sharing of injection equipment. Higher levels of psychological distress were associated with a greater likelihood of reporting drug paraphernalia sharing. These findings suggest that intervention approaches for reducing HIV-seropositive IDUs' transmission of blood-borne infections should include peer-focused interventions to alter norms of drug paraphernalia sharing and promoting primary HIV care and mental health services. PMID- 19186358 TI - Tackling patient safety. PMID- 19186359 TI - Top-ups for NHS patients: the welcomed next stage. PMID- 19186360 TI - Shape and direction for the future. PMID- 19186361 TI - Evaluating education in palliative care with link nurses in nursing homes. AB - BACKGROUND: Palliative care is recognized as an important component of care for everyone with advanced illness. Historically, it has been provided in specialist settings, but it has been suggested that best practice in palliative care should be transferred to non-specialist settings, including care homes. Care homes require particular support for this and link nurses have been recruited to develop palliative care in these settings. AIM: To assess the palliative care education received and consequently cascaded by designated nursing home staff. METHOD: Questionnaire administered to private nursing home nurses who attended a palliative care training programme in one UK region. FINDINGS: Thirty questionnaires were returned (response rate 77%). There was a high satisfaction with course content, facilitation and benefits accrued from participation. Many respondents (83%) had not commenced cascading training within their nursing homes due to lack of time and competing mandatory demands. CONCLUSION: Extending palliative care practice to non-specialist settings with the help of link nurses is possible and welcomed by nursing home staff. However, more substantive and on going support is needed post-training from both nursing home management and training facilitator to enable and empower link nurses to undertake palliative care education with their peers. PMID- 19186362 TI - Valuing local diversity in palliative care: translating the concept. AB - The contemporary challenges associated with addressing diversity, ethnicity, equality and accessibility in today's healthcare economy, sometimes lead to a reactive response where service providers strive to apply these concepts in practice. This article describes establishing a group that could engage with the broadest spectrum of the local community in ways that would make a lasting and meaningful difference to the local population, including how individuals and groups engage with and access palliative care services. The Valuing Local Diversity in Palliative Care Group was formed in May 2006. The group, whose membership is composed of statutory and voluntary services and members of various community groups, has promoted some innovative and creative partnerships. PMID- 19186363 TI - Breast cancer. Part 3: advanced cancer and psychological implications. AB - This is the last article in this 3-part series on breast cancer. The previous two articles have outlined the principles behind breast awareness and breast health, detailing common benign breast diseases, types of breast cancer and staging, and treatment for breast cancer, including surgery, chemotherapy, radiotherapy and endocrine treatment. The series concludes by giving information on advanced disease, including when a patient presents late with a fungating breast lesion, or if the disease has metastasized from the breast to other organs. Lymphoedema is also described and discussed, and the latter half of this article discusses psychological implications of breast cancer, from diagnosis through the individual treatments. PMID- 19186364 TI - Travel health. Part 2: advising travellers visiting friends and relatives abroad. AB - International travel has become more accessible and affordable, and travel, particularly to tropical and malaria regions, has increased by up to 8% annually. This change in travel has surprisingly not resulted in an increase in imported diseases. Surveillance reports of hepatitis A and enteric fever have not increased and a significant and sustained fall in malaria over the decade has been described. Nurses in primary care are the predominant providers of pre travel health services and they have an important and influential role in preventing travel-associated illness. This is the second article in a 3-part series on the spectrum of health issues associated with travel. Part one discussed pre-travel health advice, including risk assessment and educating travellers. This article explores the highest risk group of traveller, those visiting friends and relatives (VFRs). The article highlights the specific disease risks for VFRs and how these may be influenced by their health beliefs. The article explores ways in which nurses can optimize the travel health consultation to ensure that the specific needs ofVFRs are met and that they receive accurate and achievable advice. PMID- 19186365 TI - Professional regulation. Part 7a: mechanisms of government control. AB - Previous articles in this series have sought to present an historical chronology of events that have produced the healthcare environment of today. The government has been examined by reference to direct political materials and to the literature produced by the Department of Health. This article and the next aim to concentrate on a review of the current picture; of the mechanisms created by government designed to direct and oversee the regulation of certain sections of the healthcare workforce. PMID- 19186366 TI - Management of diabetes mellitus in clinical practice. AB - Diabetes is a global health problem, with a challenging epidemiology. It is one of the major health problems affecting countries around the world, particularly the UK (World Health Organization [WHO], 2006; Department of Health [DH], 2006b). It is also a growing public health threat in the US (Mokdad et al, 2001). As a result, diabetes has become an important public health issue, prompting the WHO and International Diabetes Federation (IDF) to adopt the theme 'Diabetes for Everyone' for World Diabetes Day 2006. In 2007, on World Diabetes Day, the United Nations also launched its 'Living with Diabetes at School' campaign, in response to the growing diabetes 'epidemics' (Diabetes UK, 2007). This article provides an overview of diabetes mellitus and its acute- and long-term management, including definition, aetiology, pathophysiology, classification, signs, symptoms and complications. The role of the nurse in providing patient-centred care for people with diabetes is emphasized. PMID- 19186367 TI - Understanding key issues in gender-variant children and young people. AB - Children, parents and other family members are often concerned if the behaviour of the child is more like that of children of the opposite gender, or when they show signs that they are uncomfortable with their physical sex appearance. Gender identity disorders in children and young people (those up to the age of 17 years) are rare. According to the Department of Health, they are best understood as a natural, although unusual, variation in human development. Gender variance is complex and involves psychological, biological, family and social issues. A multidisciplinary approach to care and treatment is advocated. This article provides nurses with some understanding of gender variance in children and young people in order for them to begin to help children, young people and their families. PMID- 19186368 TI - Autonomous emergency ophthalmic nursing: a pilot educational module. AB - Enhanced and extended roles proliferate in nursing today. This is, in part, due to changes in doctors' working hours and nursing becoming increasingly specialized. Ophthalmology has always had enhanced roles for nurses and the boundaries continue to evolve and widen. Accredited education in emergency ophthalmic nursing practice is essential to enhance safe practice, peer recognition and professional development in keeping with the Nursing and Midwifery Council's Code of Professional Conduct. Finding no appropriate educational and practice programme available, locally or nationally, the authors decided to devise their own module in conjunction with their ophthalmic department, their hospital's continuing professional practice development department, and Queen Margaret University. This article describes the authors' experience and the progression from inception, planning and organization to completion, and, finally, piloting the module themselves. PMID- 19186369 TI - Clothes and character: who's who on hospital wards. PMID- 19186375 TI - Effect of progestogen and progestogen type on hemostasis measures in postmenopausal women: the Postmenopausal Estrogen/Progestin Intervention (PEPI) Study. PMID- 19186376 TI - Findings of scientific misconduct. PMID- 19186377 TI - Seal a Smile program become part of a larger initiative, Growing Up Fit. PMID- 19186378 TI - A reminder: dentistry's role in preventing child abuse and neglect. PMID- 19186380 TI - The "drill" on dental tourism. Although the dental tourism trend may sound appealing at first, it can threaten your dental health. PMID- 19186379 TI - Dentists on a mission. Dentists have taken oral health to a global level- traveling to countries like Honduras, Costa Rica, Haiti and Peru to provide much needed care. PMID- 19186381 TI - The scoop on whitening treatments. PMID- 19186382 TI - Dentistry 101. PMID- 19186383 TI - What your mouth says about your body. PMID- 19186384 TI - FDA addresses access-to-care issues. PMID- 19186385 TI - Case of the month. Mucoepidermoid carcinoma. PMID- 19186386 TI - What is FLADPAC and how does it benefit me? Every year new issues arise in the legislature that affect our profession and the FDA stands ready to protect you. PMID- 19186387 TI - Case of the month. Squamous cell carcinoma. PMID- 19186388 TI - CDHC advances: workforce model pilot project gains house support. PMID- 19186389 TI - FDA dentist of the year: Dr. Donald Cadle. PMID- 19186390 TI - Making your practice visible: practical dos and don'ts of practice marketing. PMID- 19186391 TI - Women's bread -- men's capital: the domestic economy of small textile entrepreneurs in rural Zurich in the 17th and 18th centuries. PMID- 19186392 TI - Continuity and change among the Rhemish proletariat: preindustrial textile work in family perspective. PMID- 19186393 TI - Adolescent migrants from Normandy in Paris at the end of the 18th century. PMID- 19186394 TI - Orphans, apprenticeships, and the world of work: Trinite and Saint-Exprit hospitals in Paris in the 17th century. PMID- 19186395 TI - International migration policies: 1950-2000. PMID- 19186396 TI - Conflict of interest and oncology guidelines. PMID- 19186397 TI - Use of the AGREE tool will improve guideline implementation. PMID- 19186399 TI - [A disease following general anesthesia]. PMID- 19186398 TI - [A 17-year-old woman with acute abdomen]. PMID- 19186400 TI - Ethics and compassionate cancer care: where do we draw the line? PMID- 19186401 TI - Making the most of your silver dressings. PMID- 19186402 TI - Clincal snapshot. Rostral crossbite. PMID- 19186403 TI - Minimizing complications of pain and dressing adherence in the treatment of venous leg ulcers. PMID- 19186404 TI - How one woman's passion is changing international wound care. Interview by Chimere G Holmes. PMID- 19186406 TI - Electrical impedance tomography. AB - PURPOSE OF REVIEW: Electrical impedance tomography (EIT) is a noninvasive, radiation-free monitoring tool that allows real-time imaging of ventilation. The purpose of this article is to discuss the fundamentals of EIT and to review the use of EIT in critical care patients. RECENT FINDINGS: In addition to its established role in describing the distribution of alveolar ventilation, EIT has been shown to be a useful tool to detect lung collapse and monitor lung recruitment, both regionally and on a global basis. EIT has also been used to diagnose with high sensitivity incident pneumothoraces during mechanical ventilation. Additionally, with injection of hypertonic saline as a contrast agent, it is possible to estimate ventilation/perfusion distributions. SUMMARY: EIT is cheap, noninvasive and allows continuous monitoring of ventilation. It is gaining acceptance as a valuable monitoring tool for the care of critical patients. PMID- 19186407 TI - Ventilator-associated pneumonia. AB - PURPOSE OF REVIEW: Ventilator-associated pneumonia (VAP) is the main nosocomial infection in patients receiving mechanical ventilation. Despite numerous advances in the understanding of this disorder, the incidence rate continues in an unacceptable range. In this review, we discuss some important findings of recently published studies on diagnosis, prevention and treatment. RECENT FINDINGS: The microbiological sampling of the lower airways may be performed by invasive or noninvasive methods. The use of blind techniques to sample the lower respiratory tree has gained wide acceptance within the critical care setting. The use of cytological parameters such as the percentage of infected cells (cells containing phagocytised bacteria) may add objectivity to the unspecific clinical suspicion of VAP. A lot of information on the subject of prevention of VAP has been published recently that evaluates several preventive measures including new antiseptic-coated endotracheal tubes, new cuff shape and meta-analysis of known techniques. However, the clinicians must choose a bundle of measures and implement them in their intensive care units. The effectiveness of the bundles must be documented. New studies emphasize the key role of an appropriate empirical treatment. The de-escalation strategy increases the reduction of antimicrobials without worsening the outcome of VAP patients. The efficacy of monotherapy in the treatment of this infectious disease has been evaluated in new studies with controversial results. The diagnostic approach and therapy of the VAP patients are clarified with these studies. SUMMARY: In the last year, numerous articles have been published on diagnosis, treatment and prevention of VAP. In this review, we have selected those articles that potentially could lead to changes in clinical practice: Use of noninvasive techniques for diagnosis, new methods and strategies for prevention, and, finally, the efficacy of monotherapy and de-escalation in the treatment of VAP. PMID- 19186405 TI - Diffusion-weighted magnetic resonance imaging as a cancer biomarker: consensus and recommendations. AB - On May 3, 2008, a National Cancer Institute (NCI)-sponsored open consensus conference was held in Toronto, Ontario, Canada, during the 2008 International Society for Magnetic Resonance in Medicine Meeting. Approximately 100 experts and stakeholders summarized the current understanding of diffusion-weighted magnetic resonance imaging (DW-MRI) and reached consensus on the use of DW-MRI as a cancer imaging biomarker. DW-MRI should be tested as an imaging biomarker in the context of well-defined clinical trials, by adding DW-MRI to existing NCI-sponsored trials, particularly those with tissue sampling or survival indicators. Where possible, DW-MRI measurements should be compared with histologic indices including cellularity and tissue response. There is a need for tissue equivalent diffusivity phantoms; meanwhile, simple fluid-filled phantoms should be used. Monoexponential assessments of apparent diffusion coefficient values should use two b values (>100 and between 500 and 1000 mm2/sec depending on the application). Free breathing with multiple acquisitions is superior to complex gating techniques. Baseline patient reproducibility studies should be part of study designs. Both region of interest and histogram analysis of apparent diffusion coefficient measurements should be obtained. Standards for measurement, analysis, and display are needed. Annotated data from validation studies (along with outcome measures) should be made publicly available. Magnetic resonance imaging vendors should be engaged in this process. The NCI should establish a task force of experts (physicists, radiologists, and oncologists) to plan, organize technical aspects, and conduct pilot trials. The American College of Radiology Imaging Network infrastructure may be suitable for these purposes. There is an extraordinary opportunity for DW-MRI to evolve into a clinically valuable imaging tool, potentially important for drug development. PMID- 19186408 TI - Migration movements from and into Poland in the light of east-west European migration. PMID- 19186409 TI - Mechanical ventilation: epidemiological insights into current practices. AB - PURPOSE OF REVIEW: To describe the trends in the results of epidemiological studies of mechanical ventilation. RECENT FINDINGS: Changes in population demographics have increased the incidence of mechanical ventilation. Higher age and comorbidity rates portend poorer outcomes of mechanical ventilation. The most common indication for initiation of mechanical ventilation is acute respiratory failure, including postoperative respiratory failure, pneumonia, sepsis, and acute respiratory distress syndrome. Patients with sepsis and acute respiratory distress syndrome have a much higher mortality risk than the rest of this population. Changes over time in the selection of modes of ventilation, tidal volumes, positive end-expiratory pressure levels, weaning strategies, and tracheostomy timing appear to accord with data from randomized controlled trials in the literature. However, despite these changes, observational studies have not detected a statistically significant change in adjusted mortality over time. SUMMARY: The burden of critical illness will likely continue to increase in the future. Evidence from randomized trials appears to have affected the management of mechanical ventilation, but adherence to evidence-based practices may not be ideal. PMID- 19186410 TI - Paralytics in critical care: not always the bad guy. AB - PURPOSE OF REVIEW: The use of neuromuscular blocking agents (NMBAs) in patients with acute respiratory distress syndrome (ARDS) and acute lung injury remains controversial, although frequent. This review analyzes the effects of NMBAs on thoracopulmonary mechanics, gas exchange, patient outcome and their potential adverse effects. RECENT FINDINGS: NMBAs are used in 25-45% of acute lung injury/ARDS patients for a mean period of 12 days, especially in severe ARDS. Hypoxemia and facilitation of mechanical ventilation are the main indications of NMBAs. Two randomized controlled trials showed that the systematic early use of NMBAs is associated with a sustained improvement in oxygenation in ARDS patients. The most recent suggests a beneficial effect on proinflammatory response associated with ARDS and mechanical ventilation. SUMMARY: The use of NMBAs in acute lung injury/ARDS patients is not marginal. Recent studies suggest a beneficial effect of early use of NMBAs on oxygenation and inflammation. The role of NMBAs in the occurrence of ICU-acquired neuromyopathies and lung atelectasis in ARDS patients remains largely questioned. The use of NMBAs in the early phase of ARDS could reinforce the beneficial effects of a lung-protective ventilation. In this context, the effect of NMBAs on the outcome of ARDS patients must be evaluated. PMID- 19186411 TI - Acute cor pulmonale. AB - PURPOSE OF REVIEW: Acute cor pulmonale is a form of acute right heart failure produced by a sudden increase in resistance to blood flow in the pulmonary circulation, which is now rapidly recognized by bedside echocardiography. RECENT FINDINGS: In the clinical setting, acute cor pulmonale is mainly observed as a complication of massive pulmonary embolism or acute respiratory distress syndrome. In acute respiratory distress syndrome, the worsening effect of mechanical ventilation has been recently emphasized. SUMMARY: As a general rule, the treatment consists in rapidly reducing resistance to blood flow in the pulmonary circulation, obtained by a specific strategy according to etiology. PMID- 19186412 TI - Female asylum-seekers in the Netherlands: an empirical study. PMID- 19186413 TI - Rural out-migration in the drought prone areas of Ethiopia: a multilevel analysis. PMID- 19186414 TI - The drug trade, the black economy, and society in western Amazonia. PMID- 19186415 TI - Drug trafficking and consumption in China: two case studies. PMID- 19186416 TI - Evidence in the era of globalization. Abstracts of the 16th Cochrane Colloquium. October 3-7, 2008. Freiburg, Germany. PMID- 19186417 TI - Preliminary investigation of the histological effects of laser endodontic treatment on the periradicular tissues in dogs. AB - This study was conducted to determine the biological effects on the periradicular tissues of Nd:YAG laser endodontic treatment in dogs. Twelve vital teeth (24 roots) of two mongrel dogs were randomly selected to receive conventional or Nd:YAG laser endodontic treatment. Endodontic therapy was performed on the second, third, or fourth, right or left, mandibular premolars. The teeth were evaluated radiographically and histologically at 1, 15, and 30 days after endodontic treatment. The remaining teeth served as untreated controls. The results of this study revealed laser treatment causes cell necrosis in the periodontal ligament 1 day after treatment. Conventionally prepared teeth displayed no evidence of cell necrosis in the periodontal ligament. At 15 days posttreatment, both the laser and conventionally treated teeth showed increased size and numbers of osteocytes and osteoclasts in the periradicular bone. Additionally, laser-treated teeth showed signs of initial bone resorption. At 30 days posttreatment, conventionally treated teeth began to return to normal morphology. The laser-treated teeth exhibited ankylosis, cemental lysis, and major bone remodeling. PMID- 19186418 TI - An evaluation of the ability of a dentin plug to control extrusion of thermoplasticized gutta-percha. AB - Overextension of gutta-percha when using any of the thermoplasticized obturation techniques has often been mentioned as a possible deleterious yet common sequela. The purpose of this study was to test a means of controlling the extrusion of material at the apical seat of the preparation when thermoplasticized gutta percha techniques are used. Eighty human maxillary central incisors were prepared via step-back technique, in vitro, with apical stops placed 1 mm short of the actual apical foramen and the foramen was checked for patency. Teeth were divided into 10 groups, embedded in special "tooth trays," and four groups had intentional dentin plugs of 1-mm thickness placed to enhance the apical stop. A tooth tray of patent teeth and one of dentin-plugged teeth were than obturated by (a) the Obtura technique, (b) the Ultrafil technique, (c) the Touch 'n Heat technique, and (d) the Thermafil technique. Teeth were removed from the trays, extruded material carefully excised and weighed, and the weights evaluated by statistical analysis to determine results. When foramen patency was maintained, the Thermafil technique demonstrated significantly greater problems with overextension of material than did other techniques. The dentin plug served as an effective means of preventing extrusion. PMID- 19186419 TI - Immunobiological activities of bacteria isolated from the root canals of postendodontic teeth with persistent periapical lesions. AB - Although intraradicular bacteria are widely considered to be the primary etiological agents of periapical lesions, the immunobiological properties of the species in question are not adequately known. The aim of this study, therefore, was to investigate the pathobiological properties of the 10 most frequently isolated endodontopathic bacterial species. Using cellular components of the pathogens obtained by sonic extraction, we have investigated their ability to induce monocyte migration, interleukin 1 production, mitogenic responses of lymphocytes, and the polyclonal activation of B lymphocytes. It was found that all of the tested species enhanced the migration of monocytes and induced mitogenesis in B lymphocytes. The polyclonal activation of B cells and the induction of interleukin 1 by monocytes were found to be stronger in Gram negative anaerobes. Furthermore, all of the tested species excluding Bacteroides oralis were poor T cell mitogens. These findings show that a wide range of pathobiological properties are attributable to the most frequently isolated endodontic pathogens. It thus seems plausible that a battery of complex immunological responses induced by such microbes lead to the formation of persistent periapical lesions. PMID- 19186420 TI - Histological examination of paraformaldehyde-exposed gutta-percha implanted in rats. AB - Adsorption of some paraformaldehyde was noted in a previous study evaluating its sterilizing effect on gutta-percha (GP). This study examined histologically the effect of this adsorption when formaldehyde-exposed GP was implanted into the subcutaneous connective tissue of rats. GP implants were prepared in cylinder shape using a template designed to standardize size. Fifty GP cylinders were exposed to paraformaldehyde for 7 days before being implanted, while 50 others were implanted without exposure. Fifty rats had two implant sites prepared, at dorsal-interscapular and dorsal-caudal regions. Sham operations were performed on 10 rats to examine the effect of the surgery itself. The animals were killed at 4, 7, 14, 28, and 56 days. There was a significant difference between the two categories of implants only at 7 days, with the GP specimens without paraformaldehyde exposure showing more inflammation than GP with paraformaldehyde specimens (p = 0.043). Although the GP-alone specimens showed greater initial inflammation, both groups recovered in the same time period. One of the GP specimens with paraformaldehyde still showed a moderate/severe response at 56 days, whereas all of the GP-alone specimens showed only none/mild responses. The GP examined appeared to cause more inflammation than was expected. PMID- 19186421 TI - A comparison between Thermafil and lateral condensation in highly curved canals. AB - The obturation of highly curved canals often poses a problem for the clinician using lateral condensation. Recently, a product that incorporates guttapercha with a carrier (Thermafil) has been marketed. This study asked, "Was there a difference in apical microleakage of highly curved canals obturated with the Thermafil versus lateral condensation?" Twenty-two extracted human molars with canal curvature greater than 30 degrees were instrumented using a step-back technique. Teeth were placed in two groups of 10 teeth. Two teeth were controls. Canals were either obturated using sealer and lateral condensation, or sealer and the Thermafil technique. The access was closed with Cavit. Teeth were coated with wax and placed into India ink for 48 h. The wax was removed and teeth were cleared. All teeth were evaluated for linear dye penetration using a x7 viewer. A t test resulted in t = 0.137 (p > 0.05). Mean leakage for teeth obturated with the Thermafil was 1.15 mm and was 1.09 mm for lateral condensation. The Thermafil technique resulted in similar dye penetration to lateral condensation in highly curved canals. PMID- 19186422 TI - Mechanistic analysis of fluid distribution across the pulpodentin complex. AB - Fluid shifts across pulpal vessels occur during the development of pulpal inflammation. Similarly, fluid shifts occur across dentin during the application of stimuli that cause hydrodynamic activation of mechanoreceptors in the pulp. Numerous dental procedures probably also induce fluid shifts across dentin. As dentin and the pulp are located in series and function in a low compliance environment, it is probable that fluid shifts across exposed dentin cause fluid shifts across pulpal vessels. This review attempts to identify the important variables that regulate fluid distribution across vascular beds in general and discusses the special cases that exist in the pulpodentin complex. PMID- 19186423 TI - Wound healing in the tissues of the periodontium following periradicular surgery. III. The osseous excisional wound. AB - Excisional wounds were made in the maxillas and mandibles of rhesus monkeys, and the osseous wound-healing responses at postsurgical intervals ranging from 1 to 28 days were evaluated by light microscopy. The excisional defects were initially filled with a coagulum which was subsequently replaced by granulation tissue emanating from the endosteal tissues. Cortical and trabecular bone forming the wound edges was devitalized, as evidenced by an absence of osteocytes in the peripheral lacunae. At 14 days postsurgery, woven bone trabeculae occupied most of the defect, with the more superficial trabeculae in direct contact with a thick band of dense fibrous connective tissue separating the osseous defect from overlying mucosal tissues. Within the defect, new bone was deposited on devitalized bone without evidence of preceding osteoclastic activity. At 28 days, the woven bone trabeculae were more mature and a functioning periosteum was now active in repair of the cortical plate. PMID- 19186424 TI - Darkfield microscopic spirochete count in the differentiation of endodontic and periodontal abscesses. AB - A previous experimental study indicated that a darkfield microscopic spirochete count of 0 to 10% was indicative of a lesion entirely of endodontic origin and a count between 30 and 58% suggested a purely periodontal lesion. Two clinical cases are described where the differentiation of an endodontic lesion from a periodontal lesion was difficult and the traditional diagnostic indicators either could not be used or gave conflicting results. In these cases the percentage of spirochetes in the exudates of the lesions was used as an aid to diagnosis with successful treatment results. PMID- 19186425 TI - Is endodontics losing its identity in our dental schools? AB - The program presented at the Section on Endodontics of the American Association of Dental Schools Annual Meeting in March 1990 in Cincinnati focused on the issue of identity loss of endodontics in dental schools. Program participants, comprising representatives from various institutions with different educational backgrounds and experiences, were posed a series of questions on the topic. Key issues were discussed and perspectives on the future of the specialty were presented relative to dental school reorganizations and restructuring. PMID- 19186426 TI - The American Board of Endodontics--meeting the challenge. PMID- 19186427 TI - Varnishes: the effect of a second coat on apical root leakage of retrofill amalgams. AB - The root canals of 60 single-rooted human teeth were obturated with gutta-percha. Each root was then resected and randomly received one of five retrofill procedures: copal varnish before the retrofill amalgam; copal varnish before and after the retrofill amalgam; Barrier (polyamide varnish) before the retrofill amalgam; Barrier before and after the retrofill amalgam; or retrofill amalgam with no varnish. All roots were placed in methylene blue dye for 11 days, and apical dye leakage was measured. This study showed that a single coat of either varnish prior to the retrofill improves the apical seal. No statistically significant differences were observed between the use of the two varnishes or between placement of a second coat of either varnish and the use of only one coat. PMID- 19186428 TI - Ex vivo evaluation of side-perforated needles by injection of anesthetic solutions into the mandibular bone and the periodontal ligament of pigs. AB - New 27-gauge side-perforated needles were compared with conventional needles during injection in dense tissues on the basis of (a) the force to be applied on the syringe plunger and (b) the frequency of complete needle clogging. A miniaturized force transducer was used to measure the forces applied to the syringe plunger. One hundred intraosseous injections and one hundred intraligamentary injections were performed in fresh pig mandibles, using both types of needles. Results showed that (a) less force was required when performing injections with the side-perforated needles and (b) side-perforated needles got clogged less frequently than conventional ones. This result is of special interest for routine dental practice. PMID- 19186429 TI - Numerical density of dentinal tubules at the pulpal wall of human permanent premolars and third molars. AB - This study provides base line data on the density of dentinal tubules at the pulpal wall of permanent human premolars and third molars. A total of 125 healthy teeth removed for orthodontic (premolars) or other reasons (third molars) were used. After fixation the teeth were divided in the desired planes so as to obtain mesial/distal, vestibular/oral, and occlusal segments. The soft tissue was removed using 5% sodium hypochlorite (NaOCI) solution and the specimens were processed for scanning electron microscopy. Micrographs of standard magnification were taken from nine specific sites from the premolars and five from the third molars. The number of tubules present in an area representing 4000 microm2 was counted from each of the 780 micrographs, based on which the tubule density per mm2 for each tooth site was calculated. For the premolars the coronal dentin showed significantly higher tubule density than the radicular dentin. From the cementoenamel junction to mid-root level the average density of tubules declined by about one third to one half. At the level of the cementoenamel junction, the opposing mesial/distal and vestibular/oral sides showed a similar number of tubules per mm2. However, the number of tubules present on the vestibular/oral walls was significantly higher than those on the mesial/distal walls. The regional differences were less pronounced at the mid-root level of radicular dentin. For both the maxillary and mandibular third molars no significant regional difference in tubule density could be observed among the five sites examined. The mesial, distal, and occlusal walls of the mandibular third molar showed significantly greater tubule density than that of the respective walls of the maxillary third. PMID- 19186430 TI - Root canal sealer cytotoxicity with human gingival Fibroblasts. III. Calcium hydroxide-based sealers. AB - The cytotoxicity of four different calcium hydroxide-based root canal sealers (Sealapex, Apexit, CRCS, and Endoflas FS) was tested using human gingiva fibroblasts. The fibroblasts were acquired from the attached gingiva of apparently healthy patients. The sealers were allowed to set for 24 and 48 h before being covered with a fibroblastic suspension. The estimation of the incorporation rate of L-[14C]leucine in the fibroblasts after being in contact with the sealers was utilized to designate the fibroblasts' protein synthesis potential. The research was conducted over a period of 21 days. Endoflas FS induced a dramatic reduction in the protein synthesis potential of the fibroblasts in the 24-h group. In the 48-h group, Endoflas FS showed a slightly better response. Endoflas FS, however, showed a significantly higher cytotoxicity with respect to other sealers in both trials (p < 0.05). Sealapex demonstrated a relatively low cytotoxicity after 3 days of culturing. Although CRCS showed a slightly higher cytotoxicity during the initial phase of the experiments, a declining level of toxicity could be measured after 3 days of culturing. Apexit had a relatively high cytotoxicity in the beginning phase, but an ascending incorporation rate of L-[14C]leucine in the fibroblasts could be distinguished after 3 days of culturing. PMID- 19186432 TI - Healing of horizontal root fractures. AB - The roots of six anterior maxillary and mandibular teeth were intentionally fractured in three monkeys. The teeth were then rigidly splinted for 12 wk. The healing of the fractured roots was examined clinically, radiographically, and histologically for a period of 6 months. Five of six fracture sites healed by either dense connective tissue or mineralized hard tissue which appeared to enter the fracture sites from the periodontal ligament. The canal spaces in these teeth were reduced substantially by calcific deposits. The remaining pulp tissue appeared vital histologically, but with areas of odontoblastic layers absent. The fractured root which failed to heal showed presence of necrotic pulp in both apical and coronal segments and the fracture site was infiltrated by inflammatory cells. PMID- 19186431 TI - Effect of different protective base materials on hydrogen peroxide leakage during intracoronal bleaching in vitro. AB - External root resorption may develop following intracoronal bleaching with hydrogen peroxide. The preventive effect of different base materials on the radicular penetration of H2O2 during intracoronal bleaching was assessed. Seventy two bovine teeth and 20 human teeth were bleached with 30% H2O2. The bovine teeth were divided into four groups and the root canals filled with either IRM, zinc oxide-eugenol, composite resin, or glass ionomer. The radicular H2O2 penetration of each group at different layer thickness was compared. The experiment with the human teeth was performed in three stages. In the first stage no protective base was used. In the second stage IRM was placed to the cementoenamel junction level. In the third stage the IRM layer was removed 0.5 mm below the cementoenamel junction. None of the materials tested in the bovine teeth showed H2O2 penetration with a base thickness of 2 mm. When the base thickness was reduced to 1 mm, several teeth showed H2O2 penetration; however, there was no significant difference among the materials tested. When the base thickness was reduced to 0.5 mm, the H2O2 penetration in each group increased. A statistical difference was found between the composite and the glass ionomer (p < 0.05). The results for the human teeth showed that IRM layer placed at the cementoenamel junction level significantly reduced the radicular H2O2 penetration as compared with teeth where the IRM was either placed 0.5 mm below the cementoenamel junction or not placed at all (p < 0.01). It is therefore recommended that a protective base be placed to the cementoenamel junction level before intracoronal bleaching to prevent possible H2O2 hazards. PMID- 19186433 TI - Cleaning and shaping the root canal with a Nd:YAG laser beam: a comparative study. AB - The effectiveness of an experimental Nd:YAG laser fiberoptic delivery system was compared with conventional methods for its ability to cleanse and shape the root canal space in vitro. Thirty-two teeth were divided into two groups. In the first group, the canals were instrumented with K files, in the second the initial preparation was done with a K file and completed with a laser beam. Scanning electron microscopic evaluations showed that preparation with a laser beam is possible and results in an improvement in the cleanliness of the canal walls when compared with conventional techniques. PMID- 19186434 TI - Effects of carbamide peroxide on dental luting agents. AB - Vital bleaching of natural dentition using a 10% carbamide peroxide in solution may adversely effect restorative materials such as the luting agents. The purpose of this study is to examine surface structure and alterations of atomic weight percentages of elements in (a) zinc phosphate, (b) glass-ionomer, and (c) resin cements after contact with the solution. Dry samples of cement were immersed in a 10% carbamide peroxide solution (pH 4.15) for 1, 6, and 24 h and compared with controls. Observations by light microscopy, energy-dispersive X-ray microanalysis, and scanning electron microscopy were recorded. Scanning electron microscopic analysis indicated an eroded matrix in the ZnPO4 and glass ionomer samples exposed to carbamide peroxide. Energy-dispersive X-ray microanalysis data indicate a definite time-dependent decrease in the aluminum content for both cements. Surface integrity and atomic weight percentage of elements in resin cement were relatively unchanged after treatment with the peroxide solution or water. PMID- 19186435 TI - Endodontic treatment of an unusual case of fusion. AB - This case report describes the endodontic and restorative treatment of two anterior permanent teeth with unusual external and internal anatomy due to lusion. A maxillary right central incisor had fused with a supernumerary and the maxillary left central incisor had fused with the left lateral incisor. The complexity of the root canal systems in each tooth required that different endodontic obturation techniques be used. The restorative treatment involved large complex chamber-retained composite buildups and direct veneers. PMID- 19186436 TI - Zebra X. Part 1. PMID- 19186437 TI - SMI's impetus revealed. Organization strives to better health care's supply chain. Interview by Erin Burke. PMID- 19186438 TI - Trends in health care. PMID- 19186439 TI - Networking can help solve issues, educate and reinforce values. PMID- 19186440 TI - UPDATE: the Joint Commission's hospital deeming authority application. PMID- 19186441 TI - Approved: remaining component to the accreditation decision methodology for 2009. PMID- 19186442 TI - Joint Commission resources launches 2009 manual E-ditions. PMID- 19186443 TI - Approved: laboratory accreditation decisions will not affect hospital decisions. PMID- 19186444 TI - Letter from Dr. Chassin: the use of standing orders in hospitals. PMID- 19186445 TI - Approved: elimination of one-year follow-up survey for certain organizations new to the accreditation/certification process. PMID- 19186446 TI - Are rapid response teams saving lives? A new study says no. PMID- 19186447 TI - Outlying benefits of RRTs, can we discount those? PMID- 19186448 TI - To collaborate or not? Does it make a real difference? PMID- 19186449 TI - Could TJC's antibiotic standard get you sued? PMID- 19186450 TI - Matching nurses, skill sets spells ED success. PMID- 19186451 TI - Parotid surgery in patients over seventy-five years old. AB - Malignant parotid tumours are generally rare but become more common in the last decades of life; this increased incidence concerns mainly secondary parotid space involvement from metastasis or direct invasion. During the past five years, we observed and operated upon 84 patients for parotid diseases, performing 86 parotidectomies (2 bilateral operations). The patients included 30 (35.7%) over 75 years old (2 subjects over 90), mean age 80.5 years, object of the present study. In these 30 patients 2 chronic infections were observed, 6 benign tumours and 22 malignant tumours with only 6 primary neoplastic lesions. Overall, 14 extended radical parotidectomies were performed, 13 conservative total parotidectomies, 2 superficial parotidectomies and 1 radical parotidectomy, a few associated with neck dissection and/or radiotherapy. Three patients died in the post-operative period from heart attack. Five patients died from disease (3 from melanomas--2 after 3 years and 1 after 2 years--and 2 from primary carcinomas--1 after 2 years and 1 after 1 year). Six patients died without disease from various causes (2 after 4 years, the others after 3, 2, 1 year and 8 months). Nine patients are alive, 8 NED and 1 with disease in the ethmoid after 2 years, the latter well under control with radiotherapy. This experience implies that surgical treatment of these secondary tumours, which are performed more frequently in old age, exposes the patient to the risk of serious complications (3 patients died in the immediate post-operative period) and stresses the importance of careful evaluation of general conditions. However, surgery seems to have been the treatment indicated also in the patients who subsequently died from the disease, but in whom at least two-year survival was achieved in almost all cases. Unfortunately, because of the advanced age of the patients, the final results are penalized by deaths from natural causes. It is, nevertheless, encouraging that over one third of the patients operated upon are free of disease. PMID- 19186452 TI - Paraclinical evaluation of side-effects of Taxanes on auditory system. AB - Ototoxicity is one of the major causes of hearing loss and balance system disorders. Taxanes are a new group of anti-neoplastic agents used for chemotherapy; examples include Paclitaxel and Docetaxel. In this study, ototoxicity of these drugs has been evaluated in order to provide a means of adjusting the doses to avoid these complications. A prospective analytical study was carried out on 103 known cases of breast and ovarian cancer, during 2004 to 2006 (20 months), in the Otolaryngology, Head and Neck Surgery Department of Ahwaz University of Medical Sciences of Tehran. All patients (mean age 45 +/- 2.3 years) were treated with Taxanes. The first evaluation of hearing (using pure tone audiometery) was performed before starting treatment, the second in the middle of the treatment period and the last at the end of treatment. Results showed that nausea and vomiting were the most common side-effects of the drugs used. No significant side-effects of Taxanes, on the audiovestibular system, were observed. In conclusion, little information concerning the ototoxic effect of Taxanes has been reported in other studies, and, in the present investigation, no significant effect on the auditory system was found. PMID- 19186453 TI - Role of the "rooming-in" on efficacy of universal neonatal hearing screening programmes. AB - Sensorineural hearing loss is one of the most common congenital abnormalities in infants and it affects approximately one to two neonates in every 1000 births. Early identification of hearing loss in the newborn is the first step for a successful rehabilitation programme. The introduction of Otoacoustic Emission responses as a useful tool in hearing screening programmes, allowed the identification of hearing loss in the well-baby nursery and in targeted populations of the neonatal intensive care unit. Recently, a new concept of breastfeeding during hospitalization after birth has been developed. Indeed, the so-called "rooming-in" allows a mother to stay with her child in the same room, located in the nursery. This new trend has been developed to avoid any psychological adverse consequences of birth on the child-mother relationship. To enhance how "rooming-in" could affect the Universal Neonatal Hearing Screening (UNHS) programmes, an analysis has been made of the data coming from two maternity wards using different breastfeeding protocols. Data obtained demonstrate a worse performance on obtaining essential benchmark in the UNHS in the maternity ward where rooming-in is adopted (60% of newborns tested). UNHS programme efficacy could be affected by the wider adoption of the "rooming-in" regimen in the maternity wards and early detection of hearing loss revealed by UNHS could be vanished by dispersion of patients. In fact, more data are necessary to evaluate the impact of rooming, even though our data show a worsening in the UNHS results. PMID- 19186454 TI - Psychological aspects and treatment of patients with nasal septal perforation due to cocaine inhalation. AB - Use of cocaine, by inhalation, is currently increasing in Western Countries and its use is superseding heroin in the rising generation. Young people of the third millennium use narcotics to avoid the negative conditions of daily life and to escape on "unreal" trips, as happened in the '60s and '70s for the heroin addicted. Today, on the contrary, people addicted to cocaine want to be more competitive and "winners" and believe that cocaine can help them to reach this goal. A series of 104 patients (75 male, 29 female), aged between 16 and 54 yrs, all habitual inhaling cocaine users (> or = 10 times per month) have been observed for 2 years. Among them, 11 (10.5%) had nasal septal perforation, which is frequently related to cocaine use. Of these 11 patients, 8 (72.7%) had nasal septal perforation of the quadrangular cartilage, while in the other 3 (27.3%) the perforation involved also the bony tract (vomer-perpendicular ethmoidal lamina). Psychological analysis of these 104 patients is reported: 62 patients (59.6%) answered that they inhaled cocaine to improve endurance and to feel stronger and less tired; 34 patients (32.7%) in order to enjoy themselves more during parties and to communicate more effectively with other people; 5 patients (4.8%) to gain confidence and to overcome their shyness, 2 patients (1.9%) to improve their sexual performance and 1 patient (1%) to drink more alcoholic drinks for a longer time without feeling sleepy. All the patients underwent psychotherapeutic treatment, but the lack of compliance and constantly missing the scheduled follow-up visits resulted in complete therapy being performed in only 16 patients (15.3%). All the patients with nasal septal perforation underwent rhino-endoscopy, at T0, with 0 degrees, 45 degrees endoscopes, computed tomography scan of nose and paranasal sinuses and biopsy. At the time of the observational period, none of the 11 patients who presented nasal septal perforation agreed to stop cocaine abuse; therefore, a temporary solution has been offered to all the patients (accepted by 3 of them), i.e., the positioning of a silicone button to close the perforation and, thus, improve the air flow in the nose and reduce progression of local necrosis. Together with the button, the positioning is described, under local anaesthesia, of two layers per septal side of hyaluronic acid, at different levels of esterification, kept in site by the button as a "sandwich" in order to obtain better re-growth of the mucosa and fewer scabs and bleeding. PMID- 19186456 TI - Salivary gland diseases a challenge for GPs and surgeons. PMID- 19186455 TI - Simplified technique without skin flap for the bone-anchored hearing aid (BAHA) implant. AB - Aim of this report is to present a new surgical technique for the BAHA system implant and to discuss the operational techniques and complications related to this type of surgery. The common technique for the positioning of the Bone Anchored Hearing Aid (BAHA, Cochlear Limited, Englewood, CO, USA) titanium implant into the temporal bone is based on the use of either a free or a pedunculated skin flap. Reported complications of this type of surgery include skin flap necrosis with healing by second intention, infection of the flap, skin growth over the abutment, failure of osseointegration and implant extrusion. In order to reduce the incidence of these problems, different types of surgery have already been presented over the years. Herewith, a new technique is proposed for implanting a BAHA screw in the temporal bone, that is simple, rapid to perform, and does not require the use of a flap. This technique appears to offer two main advantages: i) the speeding up of the procedure; ii) the low risk of complications, such as infection and necrosis, within the skin surrounding the implant. PMID- 19186457 TI - Simultaneous pleomorphic adenoma of the parapharyngeal space and contralateral submandibular gland. Case report. AB - Herein the case is reported of a synchronous parapharyngeal space pleomorphic adenoma arising from the pharyngeal prolongation of the parotid gland and the contralateral submandibular gland, diagnosed in a young Caucasian female. Case reports and recent literature are presented. Upon physical examination, asymmetry of the lower part of the right side of the face with overlying intact skin, and a submucosal firm swelling, filling the ipsilateral side of the oropharyngeal wall, involving the right tonsillar bed which was medially displaced was immediately recognizable. The patient did not complain of dysphagia. Palpating the left submandibular region, a painless, mobile, rounded mass, 10 mm in diameter, apparently located in the submandibular gland, was detected. Magnetic resonance imaging showed that both lesions were well-defined and encapsulated. The surgical approach is discussed. Cytological diagnosis was that of a typical pleomorphic adenoma. To the best of our knowledge this is the second case report in the English literature of a concomitant pleomorphic adenoma located both in the parapharyngeal space and the submandibular gland. PMID- 19186458 TI - Giant deep lobe parotid gland pleomorphic adenoma involving the parapharyngeal space. Report of three cases and review of the diagnostic and therapeutic approaches. AB - Aim of the present report is to discuss and underline the diagnostic algorithm and the surgical approach to giant parotid pleomorphic adenomas arising in the deep lobe and growing in the parapharyngeal space. Three cases are described and a review is made of the international literature concerning giant deep lobe parotid gland pleomorphic adenoma. Diagnosis was based on imaging, computed tomography scan and magnetic resonance imaging and upon cytology, by means of fine needle aspiration biopsy. The surgical approach varied according to the location of the tumour. All patients were discharged without complications and no cases of permanent facial nerve palsy were observed. An exhaustive pre-operative diagnostic algorithm is required before approaching this lesion. Fine needle aspiration biopsy is, in our opinion, mandatory to avoid histological surprises. The surgical approach should provide excellent visibility with wide surgical exposure to secure local neurovascular structures. PMID- 19186459 TI - Parotid metastasis from renal cell carcinoma: a case report and review of the literature. AB - Renal cell carcinoma metastasis to the parotid gland after tumour nephrectomy is extremely rare. Herewith a review of the literature on this topic is discussed and a case report is presented of a 69-year-old man affected by parotid localization of renal clear cell carcinoma with neck lymph node metastases and involvement of the masseter muscle 2 years after nephrectomy. When an otolaryngologist encounters a parotid mass, diverse differential diagnoses have to be considered. A high level of suspicion of metastatic disease from the specific primary site will help in achieving correct diagnosis and evaluation of the extension of the disease. Surgical resection, even enlarged parotidectomy with neck dissection, should be considered as a therapeutic option for exclusive location of the disease in the head and neck. PMID- 19186461 TI - Coming of age: the challenges of nursing an ageing population. PMID- 19186462 TI - Partnerships and innovative models for the recruitment and retention of nurses. PMID- 19186463 TI - Smoking cessation interventions and strategies. PMID- 19186464 TI - Getting to know nurses. PMID- 19186465 TI - Hospital medical records to come under more federal scrutiny than ever. RACs, Macs, CERTs, ZPICs all will perform audits. PMID- 19186466 TI - Documentation program helps avoid revenue loss. Physician education, standardized prompts key. PMID- 19186467 TI - Discharging patients with behavioral disorders. Post-hospital follow-up is an important part. PMID- 19186468 TI - Critical Path Network. EMR triggers, concurrent review help hospital scores. Automation, checks and balances are part of the process. PMID- 19186469 TI - Critical Path Network. Easing transition from hospital to LTAC. Education of families, physicians one key to success. PMID- 19186470 TI - IP oversight required: if NV leads, will nation follow? 'Half-dozen' clinic laws under discussion. PMID- 19186472 TI - Rising water, rising anxiety. A cedar rapids agency helps the community rebuild emotionally after a devastating flood. PMID- 19186471 TI - Apply a recovery approach to wellness. Engage clients in "irresistible" services that offer them a healthier life. PMID- 19186473 TI - Everybody working together. An agency integrates housing, employment, and "traditional" treatment to help consumers better. PMID- 19186474 TI - Planning a greener future. A mental health agency plans to develop Staten Island's first green building. PMID- 19186476 TI - Ready to upgrade? PMID- 19186475 TI - Sharing the evidence. The REACH institute teaches clinicians what's working in the treatment of children. PMID- 19186477 TI - Relying on the experts. The advantages of technology assisted contracted billing services. PMID- 19186478 TI - Chronic diseases make for a perfect storm. Mental health and substance use care providers have a role in combatting them. PMID- 19186479 TI - Saving and changing lives. The new president of the Johnson Institute shares his experiences with social causes, including addiction recovery. PMID- 19186480 TI - [In the afternoon the boy was still in top condition. In the evening high fever and severe headache]. PMID- 19186481 TI - [Personal security check at the airport: how risky is the naked scan? (interview by Maria Weiss)]. PMID- 19186482 TI - [Diastolic heart failure: AT1 blocker without effect]. PMID- 19186483 TI - [INR self monitoring: large study shows no prognostic value]. PMID- 19186484 TI - [Leg venous thrombosis after work accident: serious surprise in echocardiography]. PMID- 19186485 TI - [Family physician empathy to the end: dying patients also need support]. PMID- 19186486 TI - [Hyperlipidemia in children and adolescents]. PMID- 19186487 TI - [Management of osteoarthritis]. PMID- 19186488 TI - Experts say PSOs will improve patient safety in U.S. hospitals. AB - Data collected by PSOs will not be discoverable by attorneys. Shared information about errors and causes expected to aid prevention. Many organizations will build upon proven patient safety strategies. PMID- 19186489 TI - Study: C. difficile is very common in U.S. hospitals. AB - Most high-risk patients are over 60 years of age, with multiple admissions and recent antibiotic use. Pre-emptive isolation of all patients with diarrhea is recommended. Alcohol gel is ineffective against spore form of C. difficile; soap and water help, but only bleach will kill it. PMID- 19186490 TI - Constant analysis helps improvement team succeed. AB - "It takes a good three to four months for physicians to start following the new documenation requirements", says manager. Clinical documentation specialists are aware of the case mix index, and if it changes, they drill down to understand why. Clinical documentation specialists go through six-week training period. PMID- 19186491 TI - Home visits work for behavioral health patients. AB - Whenbehavioral healthpatientswhohave been hospitalized are cared for in their home, hospital readmissions drop. PyscHealth's Home Intervention Project won a gold award from URAC. Program's goal is to increase compliance with post-hospital outpatient follow-up therapy and reduce rehospitalizations. PMID- 19186492 TI - Pre-admission prediction tool improves process. PMID- 19186493 TI - [First of all, the epidemiologist's independence is an objective condition]. PMID- 19186494 TI - [Reply to "Doubts about estimates of the number of workers at risk of lung cancer"]. PMID- 19186495 TI - [AIE position paper on waste management and health: work in progress]. PMID- 19186496 TI - [Waste management and health. Position paper of the Italian Association of Epidemiology]. PMID- 19186497 TI - ["Waste menagement and health". Letter to the Italian Association of Epidemiology]. PMID- 19186498 TI - [A comment to the study "Waste management in Campania Region: impact on health"]. PMID- 19186499 TI - [The contents of the Report: "Waste management in Campania Region: impact on health"]. PMID- 19186500 TI - [Waste management: from evidence to decision-making]. PMID- 19186501 TI - [The legislative decree n. 81/2008 on health and safety in workplaces]. PMID- 19186502 TI - [Kidney transplant and cancer risk: an epidemiological study in Northern and Central Italy]. AB - OBJECTIVE: This investigation aimed at highlighting the cancer risk of recipients of kidney transplant in northern and central Italy. METHODS: Data on 2,120 kidney transplant recipients from Niguarda Ca' Granda Hospital Milan, or from Policlinico "A. Gemelli", Rome, were analyzed The period at risk of developing cancer (person-years, PY) was computed from 30 days after transplant to date of cancer diagnosis, or date of death, or date of re-entering dialysis, or date of last follow-up. Observed and expected numbers of cancer were compared through sex and age-standardized incidence ratios (SIRs) and 95% confidence intervals (CIs). The transplant attributable fraction (AF) of cancer cases and incidence rate ratios (IRR) were also computed. RESULTS: After 16.594 PY of follow-up (median flow-up: 6.8 years), 121 cancer cases were diagnosed (729.2 cases/10(5) PY). The SIR for all cancers was 1.9. Kaposi's sarcoma (KS) (27 cases observed, SIR = 82) and non-Hodgkins lymphoma (NHL) (18 cases observed a SIR = 6.4 were the most common cancers. Significantly increased SIRs were also noted for native kidney (11 cases observed SIR = 4.9), corpus uteri (6 cases observed SIR = 4.6), and liver (6 cases observed, SIR = 3.1). The transplant AF was 46.9%, largely due to KS (98.8%) and NHL (84.3%). Since SIRs decreased with increasing age, the transplant AF ranged from 73.2% below 45 years of age to 30.4% after 54. Among risk factors, area of birth strongly influenced the risk of KS, with a 3-fold higher risk in those born in the South of Italy as compared to those born in the northern part. CONCLUSIONS: Immune depression after kidney transplantation entails a two-fold increased overall risk of cancer, mainly related to cancers associated to a viral aetiology. Furthermore, our findings suggest the need to adopt a specific serological screening for the prevention of post-transplant KS in individuals born in southern Italy. PMID- 19186503 TI - Effects of toxic oil syndrome on the psychological conditions of the descendants of affected persons. AB - OBJECTIVE: In May 1981, the Toxic Oil Syndrome (TOS) affected over 20,000 people, in Spain, as a result of the ingestion of rapeseed oil that had been denatured with 2% aniline. Amongst many physical and organic problems, many patients in this cohort showed different degrees of anxiety and depression. It can be hypothesized that their children might well be susceptible to suffer from anxiety, depression and other psychological disturbances. METHODS: Children with a father and/or mother included in the official TOS census, who were born between 1st January 1983 and 31st December 1989 and resided in Madrid (n. 420, response rate 84%), were compared against high-school children of TOS-free parents of the same age and similar socioeconomic status (n. 327). DATA COLLECTION: Spanish version of Goldberg and Hillier's General Health Questionnaire (GHQ-60) and Cattell's High School Personality Questionnaire (HSPQ). RESULTS: The only statistically significant difference between the two groups was the sleep disturbance factor of the GHQ-60 questionnaire. Significant differences were not observed in any of the personality factors (such as anxiety, depression, excitability and introversion) analysed by the HSPQ questionnaire when the exposed group was considered as a whole. However, in the replies to the HSPQ questionnaire, some statistically significant differences between exposed and non exposed children were detected in analyses carried out separately in each sex. CONCLUSIONS: The results of this study tend to rule out any impairment of the mental health of children born from parents who had been TOS victims. PMID- 19186504 TI - [Mortality in the cohort of the asbestos cement workers in the Eternit plant in Casale Monferrato (Italy)]. AB - OBJECTIVE: The present report updates the mortality cohort study of "Eternit" workers in Casale Monferrato, one of the major plant for the production of corrugated and plain sheets, tubes and high-pressure pipes in asbestos-cement in Italy active between 1907-1986. DESIGN: Retrospective cohort study. SETTING: The cohort included 3,443 blue-collar workers (2,663 men and 780 women) active between 1950 and 1986. Nine subjects had been excluded because of incomplete data. The follow-up ended in 2003. Mortality in the cohort was compared to the cause-specific rates for age and period of the Piedmont population and the SMR values were calculated for the period 1965-2003. MAIN OUTCOME: Analysis of cause specific mortality. RESULTS: Follow-up was completed for 99.0% of the subjects. At the end of follow-up, 42% of the subjects were alive, 57% deceased and 1% lost to follow-up or moved abroad. In both sexes, a statistically significant increase of morality for all causes (men: SMR 135.1; 1,438 obs vs 1,064.1 exp; p < 0.01; women: RSM 149.5; 371 obs vs 248.2 exp; p < 0.01) was observed. The SMRs for lung, pleural and peritoneal cancer and for asbestosis increased according to duration of exposure and latency. For pleural cancer, increasing risks at shorter latencies were observed as exposure length increased. In men, the increase in the SMRs for lung and pleural cancer was reduced in the category of longest latency; still increased but declining SMRs were also observed at longer time since first exposure. Mortality from peritoneal cancer and asbestosis on the contrary increased with latency and with time since last exposure. Among women, a significant increase of mortality for uterine cancer (SMR 2569; 15 obs vs 5.8 exp; p < 0.01), ovarian cancer (SMR 227.3; 9 obs vs 4.0 exp; p < 0.05) and rectum cancer (SMR 318.6; 9 obs vs 2.8 exp; p < 0.01) was observed. CONCLUSION: a significant increase in mortality from the main asbestos-related diseases was confirmed by duration of exposure. In relation to latency The SMRs for lung and pleural cancer present a curvilinear trend with a decrease for longest latency periods (after 30 years from the cessation of exposure). The SMRs for peritoneal cancer and asbestosis showed a monotonic increase. PMID- 19186505 TI - [Mortality in areas with different socioeconomic characteristics in Palermo]. AB - OBJECTIVE: To assess whether mortality rates differ among areas with different values of a socioeconomic index. DESIGN: Ecological mortality study; mortality rates were compared among four areas joining census tracts categorized by quartiles of a socioeconomic status index. This index was calculated using individual 2001 census data in a factorial analysis. SETTING: The study was conducted in Palermo (Italy) a city with 668,996 inhabitants in 2006. MAIN OUTCOMES MEASURES: Sex, age group, cause specific standardized mortality rates and rate ratios, expected life years. RESULTS: The area with the lowest value of the socioeconomic index had mortality rates higher than the area with the uppermost value. This occurred in both sexes, all age groups and for most mortality causes (with few exceptions): not all rate ratios were always statistically significant. CONCLUSION: Despite limitations due to the use of an area-based index, results suggest that the welfare system is not successful in compensating health inequalities caused by socioeconomic inequalities. PMID- 19186506 TI - [Screening and vaccine: toward an integrated cervical cancer prevention programme]. AB - The identification of the human papilloma virus (HPV) as the necessary cause of cervical cancer introduced two new tools for prevention: HPV DNA test and vaccine. In this paper we analyse the plausible changes that will happens in the cervical cancer prevention in Italy and what we will need to have a governance of these changes. From a public health point of view, screening and vaccination are interventions directed to the whole community and their goal is to reach high coverage of the target population. The vaccinated women should be screened with different algorithms from the non-vaccinated ones, consequently we need individual information about the vaccination status of the women that we are going to invite for screening. Vaccine and screening interventions must be integrated in a unique public health program and screening should use less aggressive protocols, to be less expensive and more sustainable and efficient. The challenge for the Health Service is to integrate the primary and secondary prevention avoiding that one will diminish the benefits of the other and that the introduction of new technologies will be only the opportunity to double the costs with minimal health benefits. PMID- 19186507 TI - [Review on environmental impact of solid wastes produced by municipal urban waste incinerators]. AB - Incinerators reduce urban waste volumes and recover energy by combustion, but produce important quantities of solid wastes: bottom ashes and fly ashes. The presence of toxic heavy metals and persistent organic compounds, both in bottom and in fly ash, require attention for their disposal and for proposed use in cement production or as filling material in road construction. Recent ecotoxicological studies on leachates obtained from ashes produced by urban waste incinerators suggest an urgent proposal of common more stringent regulations in all Union European countries for ashes disposal and use. PMID- 19186508 TI - [Methodological issues and new developments in genetic association studies]. AB - Genetic factors are involved in the etiology of many common diseases, even if it is difficult to correctly identify the responsible genetic variations. The main goal of genetic association studies is to identify polymorphisms that vary systematically between people with different conditions of disease. Recent advances in time and cost for genotyping have rapidly moved from a candidate gene approach to genome-wide studies, but the methodological issues remain the same and furthermore increased. One of the major problem is that the genome is large and includes many polymorphic variants which could be found associated by chance. In addiction, complex diseases will require complex analyses in which many variants are assessed simultaneously. To help to distinguish causal associations from spurious ones, restrictive standard for statistical significance or considering polymorphisms which could be a-priori causally associated to disease are needed. PMID- 19186509 TI - [Chikungunya emergency in Emilia-Romagna: learning through experience]. AB - This paper summarizes the Emilia-Romagna strategy to face the 2007 emergency, caused by a Chikungunya epidemic outbreak with local virus transmission by Ae. albopictus. The paper describes the trend of epidemic and the interventions adopted to face toward the event. The first cases were in Ravenna and Cervia and then the outbreak spread toward other areas: the Provinces of Forli-Cesena, Rimini and Bologna. Last case was notified 2007 28th September; Health Ministry declared over the outbreak on 2007 20th November. Emilia-Romagna Region did not consider over the trouble and prepared a Plan for the fight against the asian tiger mosquito and the prevention of Chikungunya and Dengue fever for 2008, activating an health surveillance system and optimizing the entomological control of the territory. PMID- 19186510 TI - [Numbers. Cancer survival in Italy is within the European average]. PMID- 19186511 TI - [Biologically active compounds in the mechanism of formation of reflux nephropathy in children]. AB - A concept is proposed laying emphasis on the involvement of cytokines, growth factors, prostaglandins, and nitrogen oxides in the mechanism of formation of reflux-nephropathy in children. It is shown that enhanced production of mediators of inflammation and humoral growth factors in patients with vesico-ureter reflux depends on the degree of infiltrative and sclerotic changes in the kidneys. The accompanying dysbalance of pro- and anti-inflammatory cytokines due to increased endogeneous production of TNF-alpha and concomitant decrease of daily BL-10 excretion constitutes an unfavourable prognostic factor of reflux-nephropathy. There is evidence that inclusion of ACE inhibitors (Enap) in therapy of vesico ureter reflux during 6 months leads to a marked improvement of the structural and functional state of the kidneys. It suggests high renoprotective potential of these preparations and the possibility of their use for prophylaxis of nephrosclerosis in children. PMID- 19186512 TI - [Current problems of immunoprophylaxis of infectious and non-infectious diseases]. AB - Progress in immunology greatly contributed to the understanding of mechanisms of infectious immunity and vaccine action. Moreover, it facilitated the development of new vaccines and vaccination not only of healthy children but also of subjects with health problems and chronic diseases including the use of several vaccines. The number of contraindications could be decreased without a rise in the frequency of post-vaccination complications. Immunoprophylaxis allowed the incidence of controllable infections to be reduced; the level of epidemiologic well-being achieved in the course of this work provided a basis for eradication of some diseases. At the same time, successes of vaccination made mankind dependent on the use of vaccines. As a result, discontinuation of mass vaccination and even temporal decrease of its scale in the absence of certain infections or in the situation of their sporadic occurrence (i.e. the lack of natural immunization) lead to their reappearance. Specific features of modern immunoprophylaxis of infectious and non-infectious pathology and its timetable are described. PMID- 19186513 TI - [Bronchial asthma in children and adolescents]. AB - Modern concept of the development of bronchial asthma (BA) is discussed based on original and reported findings with reference to specific features of its etiology, pathogenesis, clinical course, and drug therapy. Special emphasis is laid on the methods of allergen-specific immunotherapy of atopic BA. Up-to-date staged treatment of different forms of BA in children by inhalation of glucocorticosteroids is described. PMID- 19186514 TI - [Modern technologies for the management of atopic dermatitis in children]. AB - Pathogenetic mechanisms of atopic dermatitis in children, its clinical features, medicamentous treatment, and modern technologies for external therapy are considered based on the results of original studies and literature data. The analysis is focused on the available experience with the use of modern dermocosmetic preparations. A strategy of stage-by-stage therapy of atopic dermatitis in children using these preparations is proposed. PMID- 19186516 TI - [Problems and prospects of care for newborn babies with an extremely low body weight]. AB - The paper is designed to discuss problems of morbidity, treatment, rehabilitation, and late follow-up development in children of extremely low birth weight (500-999 g). Despite the use of modern methods of rehabilitation and nursing, the overwhelming majority of such children develop severe combined somato-neurologic pathology leading to disability (polymorphic lesions in the nervous system, broncho-pulmonary dysplasia, retinopathy, etc.). Efficacious approach to the reduction of frequency and severity of polysystemic pathological conditions requires improvement and development of new methods of resuscitation and intensive therapy based on up-to-date neuroprotective technologies (including the use of anticytokines and stem cells) and their combination with non medicamentous therapeutic modalities. PMID- 19186515 TI - [Immediate and late results of combined treatment of dilatated cardiomyopathy in children]. AB - Effects of combined treatment of chronic heart failure on the clinical course and outcome of dilatated cardiomyopathy was evaluated in 151 children followed up for 6 months-14 years (4.0 +/- 0.5 yr). It was shown that the use of ACE inhibitors and beta-adrenoblockers in combined therapy of this pathology improved survival and quality of life of the children PMID- 19186517 TI - [General ways of improvement in dispensary dental care in the Armed Forces of the Russian Federation in the peace time]. AB - The article presents the discussions of todays' existing problems of delivery of ambulatory dental care in military-medical institutes of Ministry of Defense. The abovementioned problems are not the single ones in delivery of ambulatory dental care. However, fast solving even of pert of these problems contributes to improvement of delivery of ambulatory dental care in the Armed Forces of RF. PMID- 19186518 TI - [Mathematical analysis of the effectiveness of using bed capacity as a step in planning military hospital activities]. PMID- 19186519 TI - [Modern technologies for improvement of quality medical care administration in otorhinolaryngology]. PMID- 19186521 TI - [Physical therapy in a military medical service unit]. PMID- 19186520 TI - [Donorship--a state's affaire]. PMID- 19186522 TI - [Pathology of lungs by wounded in the armed conflicts in Northern Caucasus]. AB - According to 7000 patient histories, wounded in armed conflicts in Northern Caucasus, for the last years, is marked a big part of wounds of mild (45.3%) and medium (31.5%) severity. Heavy and extremely heavy wounds marked conformably in 20.8 and 2.4 percents. An acute respiratory distress-syndrome is marked by 32 wounded with heavy and extremely heavy wounds, including 10--with penetrating wound of brainpan, 9--penetrating wound of bosom and tense hemopneumothorax, 11- penetrating wound of abdomen (with deflection of hollow and parenchymal organs), 2--penetrating wound of lower limbs (with firing breakup of long bones and deflections of great vessel), attended by blood loss and shock. Pneumonia was found during first 3 days after the wound by 47%, on 4-7 day--31%, during second week--13%, latter--9%. In 93% of cases pneumonia was marked in combination with traumatic shock. Radiological examining of radiological plays a conclusive account in diagnostics, and according to meanings--computerized tomography. Estimation of function of exterior breathing is also important. PMID- 19186523 TI - [Comparative analyses of surgical treatment of brain aneurysms]. PMID- 19186524 TI - [Age aspects of clinical-morphological characteristics and treatment of duodenal ulcer]. PMID- 19186525 TI - [Contemporary views on non-specific ulcerative colitis and conservative methods of its treatment ]. PMID- 19186526 TI - [Modern principles of prevention of human transmitted infections]. AB - According to information about ecology of agents of parasitic diseases with lousy flea mechanism of contamination, mite natural diseases and experience of battle with them, we can make a conclusion: elimination of spongers in their natural environment, lousies and fleas by washing and disinfection is not effective. Alternative is implementation of insectoacaricides of new class, based on permethrins. These insectoacaricides are secure for environment and very effective towards arthopods. Method of non-termal preventing suction of arthopods is probed in military practice and other treatment-prophylaxis institutes. It is proposed to implement new insectoacaricides in form of spray cans as part of individual contrebiological drug-locker of every service man, acting in difficult epidemiological situation, caused by virus threat of lousy or flea infection, or by beforehanded provision by impregnated uniform or clothes. PMID- 19186527 TI - [Transient personality disorders in modern military psychiatry (review of literature)]. PMID- 19186528 TI - [The 50th anniversary of the hospital of a spaceport "Plesetsk"]. PMID- 19186529 TI - [Remodelling of peripheral vessels in cardiovascular diseases: pathogenesis and treatment with telmisartan]. AB - The authors present remodelling mechanisms of distal peripheral vessels in vibration disease, caused by endothelium functions disorder (lower NO, increased endothelin-1 and Villebrandt factor), stressed oxidative metabolism in neutrophils, intensified lipid peroxidation, depressed antioxidant defence, activated anti-inflammatory cytokines (TNF-alpha, IL-1beta) and hyperhomocysteinemia. Degree of the changes increases with combination of vibration disease and arterial hypertension. The data justify application of Telmisartan improving functional state of endothelium. PMID- 19186530 TI - [Risk factors and rehabilitation of workers with COPD in major industrial enterprise]. AB - The authors present epidemiologic characteristics of COLD in workers of major atomic machinery enterprise. The article covers high prevalence of this disease on the enterprise, evaluation of connection between occupational and non occupational risk factors and COLD development. Constant broncholytic therapy in the occupational health office caused significant improvement of clinical and functional parameters and decrease in transitory disablement duration. PMID- 19186531 TI - [Industrially mediated hemopathies and occupational diseases of blood]. AB - The article covers criteria and classification of industrially mediated hematologic changes and occupational diseases caused by occupational factors hazardous for blood, mechanisms and clinical and laboratory peculiarities of hemopathies and toxic hemopoietic disorders. The authors present diagnostic parameters for the most prevalent pathologic hemopoietic conditions. PMID- 19186532 TI - [Contemporary rehabilitation technologies for occupations at risk of vibration disease]. AB - The authors demonstrate efficiency of nitrogenous silicon baths in "Belokurikha" resort with precasted factors for neurohormonal state of patients with I degree vibration disease, for their life quality and psychoemotional state, through lower dysbalance of stimulating and peripheral hormons, lower thyroxin/triiodothyronine and estradiol/ testosteron ratios, aldosteron level, positive influence on prooxidant and antioxidant activities of blood serum, on platelet and vascular hemostasis. PMID- 19186533 TI - [On functional heart disorders in individuals with occupational diseases caused by physical and chemical factors]. AB - Heterometric mechanism of cardiac autoregulation is characteristic for patients with occupational vibration disease, as an example of physical factor in occupational disease pathogenesis. For chemical factor inducing occupational disease, as exemplified by chronic intoxication with organic solvents, homeometric mechanism of cardiac autoregulation is characteristic. PMID- 19186534 TI - [Pathogenesis, clinical and functional characteristics of chronic stress in military men]. AB - The authors demonstrate that arterial hypertension in military men serving in extreme conditions is characterized by systolo-diastolic and diastolic variants, with excessive decrease of blood pressure in night hours, prevalent albuminuria and higher values of cellular content of blood. PMID- 19186535 TI - [Medical service system for hazardous production workers in major industrial city]. AB - Inadequate training of specialists participating in periodic medical examinations remains one of the major problems of medical service for hazardous production workers, as shown by the authors. Suggestions are interactions between medical institution participating in periodic medical examinations and local medical institutions (licensed for periodic medical examinations) through setting an occupational therapist in the latter institutions for continuous observation over individuals having occupational exposure to hazards and for referral to Regional occupational pathology center, if necessary. This organizational pattern is a system of correlated structural elements, providing effective mechanism to save labour resources and more cost-effective, vs. the old pattern. PMID- 19186536 TI - [Importance of molecular genetic markers for evaluating individual risk of visceropathies in workers exposed to dust]. AB - The authors present results of evaluating molecular genetic markers to estimate individual risk and forecast visceropathies development in workers exposed to high concentrations of industrial dust aerosols. PMID- 19186537 TI - [Differentiantion of Listeria monocytogenes strains isolated in the Far East and European part of Russia on the basis of polymorphism of genes encoding invasion factors]. AB - Forty Listeria monocytogenes isolates obtained in European and Far East regions of Russia were differentiated on the basis of polymorphism of 5 markergenes. Total length of concatemers obtained after sequencing of internal fragments of genes inlA, inlB, inlC, inlE and prs was 3029 b.p. Comparative analysis of concatemers' sequences revealed 237 variable nucleotides. Totally, 25 sequence types were revealed, and isolates from European and Far East regions belonged to different types. On the dendrogram isolates split on 2 clusters, which correspond to early described phylogenetic lines of L. monocytogenes specie. Isolates obtained in European and Far East regions formed independent subclusters within main clusters. Fifteen clinical isolates of L. monocytogenes belonged to 7 different types. Analysis of epidemiologic data on time and place of isolates obtaining suggested that isolates of the same sequence type are epidemiologically related and might represent one strain; index of discrimination for proposed typing method was calculated as 0.982. PMID- 19186538 TI - [Sanitary and epidemiologic welfare of the environment and children's health]. PMID- 19186539 TI - [Value of enzyme-linked immunosorbent assay for detection of p24 antigen of human immunodeficiency virus in confirmation of HIV-infection]. AB - AIM: To evaluate diagnostic value of p24 antigen detection for algorithm of confirmatory diagnostics of HIV-infection. MATERIALS AND METHODS: Concurrently with Western blot assay (WB, "New Lav Blot1", Bio-Rad), tests for detection of p24 antigen of HIV (Genetic Systems HIV-1 Ag EIA", "VectoHIV-1 p24-antigen confirming test", and "DS-EIA-HIV-AG-SCREEN") were used for confirmation of first positive result of immuno-enzyme assay. RESULTS: p24 HIV antigen was detected in serum samples in 8.4% of patients with equivocal result of WB and in 4.2% of patients with negative and positive results of WB. Presence of p24 was correlated with high viral load, and, in patients with confirmed diagnosis, with low CD4 cells count (<500 cells/ml). p24 was detected in more than 30% of persons with confirmed seroconversion after primary testing. CONCLUSION: In groups of persons with negative and equivocal results of WB assay, detection of HIV p24 antigen points to the presence of infection and could be the reason for the final diagnosis. Detection of p24 in patients with positive result of WB assay allows to consider them as probable candidates for highly active antiretroviral therapy. PMID- 19186540 TI - [Diagnostic value of IgG, IgA, and IgM to Bordetella pertussis antigens in patients with pertussis]. AB - Comparative study of IgG, IgA, and IgM levels to complex of antigens (CA) of vaccine strain No. 475 and separate antigens of Bordetella pertussis: pertussis toxin (PT), filamentous hemagglutinin (FHA), lypopolysaccharide (LPS), agglutinogens 1 (Aggl.1) and 2 (Aggl.2) was performed by ELISA in 80 patients with pertussis and 80 healthy vaccinated children. Antibodies to mentioned antigens were detected both in ill and healthy children but their levels were remarkably higher in patients. The most reliable serologic marker of pertussis was IgA, which were detected in the majority of patients. Detection rates of this class of antibodies to CA, PT, FHA, LPS, Aggl.1, and Aggl.2 were 91%, 77.5%, 69%, 59%, 80%, and 12%, respectively. Elevated levels of specific IgA were registered in 5% of healthy children. Obtained results showed high information value of detection of the IgA and IgG antibodies to CA, PT, FHA, and Aggl.1 using ELISA for pertussis diagnostics. Simplicity and economy of the CA obtainment allow to recommend CA-based ELISA for serologic diagnostics of pertussis. PMID- 19186541 TI - [Immunization of adult population of Russia against diphtheria in 2006-2007]. AB - The vaccination coverage against diphtheria in the adult population of Russia in 2006-2007 was analyzed in different age, professional and social groups and reasons of absent immunization. Vaccination coverage in adults was 95.3%, of them 92.5% were boosted (i.e. completely immunized) and only 7.5% were in process of vaccination. Vaccination coverage was high in the majority of regions. Mean vaccination coverage in adults aged 25-49 ages was 96.2-96.5% whereas in adults > or =50 years it was lower (91.8-94.8%). Mean coverage in groups of high risk for severe diphtheria was 97.7% in professional high-risk groups and somewhat lower in social high-risk groups (94.4%). The lowest vaccination rate was registered in group of migrants (81.7%). In unvaccinated persons from professional groups 33.1% were not vaccinated due to medical contraindications, 22.8%--because of vaccination refusal, and 38.1%--due to non-medical reasons. In social groups these figures were 21.8%, 28.4%, and 49.8% respectively. Reported mean adult vaccination coverage in Russia is confirmed by results of serological studies. Protective titers were detected in 91.3% tested adults. However in the group aged > or =50 years rate of protected persons was lower (84-85%). PMID- 19186542 TI - [Influence of chitosan on immunophenotype and functional activity of murine mononuclear leukocytes after immunization with inactivated influenza vaccine]. AB - In the overwhelming majority of countries inactivated vaccines, which form mainly humoral immunity, are used for prevention of influenza. The objective of the study was to assess the combined effect of inactivated influenza vaccine and chitozan on cellular immunity in CBA line mice. Intramuscular administration of 2 doses (with 4 week interval) of inactivated influenza vaccine and chitozan resulted in increased cytotoxic activity of splenic NK cells against NK-sensitive cell line K562 as well as in increased proliferative activity of mononuclear leukocytes, and numbers of CD3 T-lymphocytes, NKT cells, B-lymphocytes in animals' spleens. Combination of inactivated influenza vaccine with chitozan modulated the number MHC II-expressing cells by eliminating the increased reactivity of immune system cells as well as increased the number of MHC I expressing cells. This point on the activation of cellular properties, which recognize intracellular pathogens, and thus on activation of both humoral and cellular factors of immune response. It can be proposed that inclusion of chitozan in the vaccine allows to modulate switching of the immune response from Th-2 to Th-1 type. PMID- 19186543 TI - [Inhibiting effect of complex of natural cytokines and cationic antimicrobial peptides on the growth of Mycobacterium tuberculosis H37Rv in vitro]. AB - AIM: To assess direct antimicrobial effect of complex of natural cytokines (CNC) and antimicrobial peptides (Syperlymph preparation; CNC) on Mycobacterium tuberculosis H37Rv (H37Rv) and effect mediated by macrophages (MP) treated with the preparation. MATERIALS AND METHODS: Direct effect of CNC was studied during cultivation of H37Rv in the presence of preparation, whereas indirect effects- during simultaneous cultivation of H37Rv and mice peritoneal MP C57B1/6. Assessment of growth was performed on the 7th day using PCR. RESULTS: It was shown that CNC directly inhibits growth of H37Rv in vitro. Cultivation of H37Rv in culture of MP resulted in inhibition of M. tuberculosis. The most evident inhibition was noted after extension of time of preliminary treatment of MP with Syperlymph and simultaneous increase of its concentration. CONCLUSION: Antimycobacterial effect of Syperlymph preparation related to complex effect of cytokines and antimicrobial peptides directed to M. tuberculosis and macrophages, which forms the conditions for killing of mycobacteria. PMID- 19186544 TI - [Experimental study of the phenomenon of antibody dependent tick-borne encephalitis virus infectivity enhancement in vitro]. AB - Phenomenon of antibody dependent tick-borne encephalitis virus (TBEV) infectivity enhancement in the human monocytic cell culture J-96 has been studied. Three specific commercial sera that are generally used for therapy and prevention of TBE in humans were used. Results showed that enhancement of TBEV infectivity was markedly apparent following use of 2 out of 3 studied sera. Data obtained in the study prove that phenomenon of antibody dependent TBEV infectivity enhancement in monocytes cell culture may exist, but conditions for its realization in the organism are very unlikely due to the narrow range of antibodies' concentration and quantity of viral particles. A the same time, the fact of possible aggravation of TBE course after use of specific antisera for treatment and prevention of TBE in humans should not be ignored. PMID- 19186545 TI - [Modulating effect of serotonin on the development of human leukocytes apoptosis induced by Yersinia]. AB - Effect of biogenic amine serotonin on the development of human blood leukocytes during interaction with species from Yersinia genus (Y. pestis EV, Y. pseudotuberculosis serovars I and IV, Y. enterocolitica serovars 09 and 03) was studied in model system in vitro using flow cytofluorometry. Serotonin in concentration 10(-5) M had differently marked effects on Yersinia spp.-induced apoptosis of leukocytes. Pattern of the observed changes depended from species and serovar of Yersinia. Serotonin inhibited development of early (after 6 hours) apoptosis of leukocytes induced by Y. pestis and Y. pseudotuberculosis serovar I. PMID- 19186546 TI - [Identification of heterologous antitoxin in sera of patients with diphtheria]. AB - Using immobilized diphtheria toxin and peroxidase conjugate of monoclonal antibodies to light chains of equine immunoglobulin a method of quantification of equine antibodies against diphtheria in sera of patients after serotherapy was developed. The sensitivity of indirect enzyme-linked immunosorbent assay was 0.0005 IU/ml, and coefficient of variation did not exceed 10%. It was shown that in patients with toxic diphtheria heterologous antitoxin is eliminated within 4-6 weeks. Level of anti-diphtheria immunoglobulin under the similar severity of disease and dosage of antitoxin can vary in wide ranges and depends from individual's characteristics. PMID- 19186547 TI - [Oral cavity microbiocenosis in healthy adolescents and patients with chronic gastroduodenitis]. AB - AIM: To determine characteristics of microbial associations and degree of oral cavity colonization by different biotopes in healthy adolescents and patients with chronic gastroduodenitis. MATERIALS AND METHODS: Saliva, dental deposit, content of tooth-gingival cavity and material from buccal and lingual mucous surfaces of 24 healthy adolescents and 29 patients of the same age with gastroduodenitis were used for bacteriological analysis. Inoculation were made on the following growth media: yolk-salt agar, Endo, Saburo, MRS as well as on Columbia and Saedler blood agars. RESULTS: It was shown that rate of isolation of peptostreptococci from oral cavity of patients was 90% lower compared with controls; rate of isolation of lactobacilli, Veillonella, and stomatococci was 30% lower, whereas hemolytic Porphyromonas, staphylococci and streptococci were isolated 20% more frequently. Concentration of peptostreptococci and streptococci was 7.0 lg CFU/ml, whereas concentration of Porphyromonas, Veillonella, peptococci and staphylococci was 6.0 lg CFU/ml, Neisseria--5.0 lg CFU/ml, stomatococci, bacilli and Candida fungi --4.5 lg CFU/ml. Decrease of concentration of lactobacilli to 4.0 lg CFU/ml was noted. CONCLUSION: Compared with healthy group of children, widened spectrum of microorganisms in all studied biotopes of oral cavity was observed in patients with chronic gastroduodenitis; species belonging to pathogenic microflora were detected in patients more frequently and in more quantities. PMID- 19186548 TI - [Biological properties of heat-labile lethal toxin of Yersinia pseudotuberculosis]. AB - The aim of the study was to assess the biological properties of heat-labile lethal protein toxin of Y. pseudotuberculosis. Toxin was extracted from Y. pseudotuberculosis strain 2517 type III serovar pYV-. The toxin killed mice 1-3 days after intraperitoneal administration (LD50=0.3 mcg of the protein). Heating at 56 degrees C during 30 min inactivated lethal activity of the toxin. It had a dose-dependent dermonecrotic effect during intracutaneous administration in rabbits. Hyperimmune rabbit serum to the toxin was obtained. Incubation of the toxin (LD100=1.2 mcg of the protein) with the serum at 37 degrees C during 30 min resulted in neutralization of lethal and dermonecrotic effects. The toxin did not have the cytotoxic effect on HeLa, Hep-2, and SPEV cells, but showed hemolytic activity to human and animal erythrocytes, and weak mitogenic activity to splenic cells of CBA line mice compared with control mitogen (concanavalin A). PMID- 19186549 TI - [Influence of thermolabile lethal toxin of Yersinia pseudotuberculosis on development of sea urchin embryos and biosynthesis of DNA and RNA in embryonic cells]. AB - Influence of thermolabile lethal toxin of Y. pseudotuberculosis on the development of embryos of sea urchin (Strongylocentrotus intermedius) and on biosynthesis of nucleic acids in embryonic cells was studied. Thermolabile lethal toxin affected metabolic processes of cells by inhibiting DNA and RNA synthesis. It had damaging action on developing embryos of sea urchin causing morphological changes and, as a consequence, death of embryos. PMID- 19186550 TI - [Development and use of a new growth medium for detection and identification of sanitary-indicative microorganisms]. AB - Threat of emergence and spreading of dangerous intestinal infections determines the necessity to control for water quality in its sources in respect of sanitary indicative microorganisms, thermo-tolerant coliforms and lactose-negative intestinal bacilli testifying fecal pollution. Standard techniques for isolation of enterobacteria are based on use of lactose-containing inhibitory and non inhibitory growth media such as lactase-peptone and Kessler media. Development of standard, effective, and reliable for use in laboratory conditions accumulation medium for detection of enterobacteria aimed to increase reliability of sanitary bacteriological monitoring of objects of aquatic environment. Two variants of Eikman medium with an added indicator were designed for differentiation of enterobacteria on the basis of lactose and glucose fermentation with application of a thermo-tolerance test at 44-46 degrees C. Maximal accumulation of coliforms on the developed media in static cultivation conditions and at sensitivity on the level of individual cells was observed after 16 +/- 2 hours, whereas in control inhibitory media--after 22 +/- 2 hours. Studies on sterilization of the developed media in disposable packages byy-irradiation were conducted to elucidate the possibility of their use in field conditions performing analysis in flasks for preservation of high growth properties. In order to reduce time for analysis on confirmation of detection of thermotolerant coliforms, addition of tryptophan in composition of the media was provided, which allows to perform an indole test and definitively confirm the detection of Escherichia coli. Pilot industrial samples of designed media were successively passed the approbation. PMID- 19186551 TI - [Comparative clinical and epidemiologic characteristic of HIV infection in women from discordant pairs]. AB - Forty HIV-positive women living with HIV-infected sexual partner as well as 40 HIV-positive women from discordant pairs (HIV-positive wife and HIV-negative husband) were surveyed. Less than half of HIV-infected patients (40%) specified the use of condoms for sexual contacts. In the group of women living with HIV infected partner more expressed immunosupression (CD4 count 427 cells/mcl versus 528 cells/mcl) was observed. Statistically significant increased viral load in the 1st group compared with the women from discordant pairs was revealed. PMID- 19186552 TI - [Microflora of pharyngeal mucosa in children with solid tumors]. AB - Microbiological study of pharyngeal mucosa in 43 children with solid tumors revealed that 77.2% of isolated microorganisms belonged to Gram-positive flora. It was shown that streptococci and Staphylococcus aureus were the main species. Species composition of streptococci included both pyogenic (S. pyogenes, S. agalactiae, S. dysgalactiae, S. equi) andviridans species (S. acidominimus, S. oralis and "S. milleri" group). Nocardioform actinomycetes, corynebacteria and other staphylococci were referred to additional microflora. Accidental microflora was represented by Neisseria spp., non-fermenting Gram-negative bacteria, enterobacteria and yeast-like fungi. Microbiologic study of pharyngeal mucosa biocenosis showed that monoculture was present only in 2.3% of cases; in other cases microorganisms formed both intra-genus and inter-species associations. 2-6 component associations were revealed with predominance of 3-4-component associations (37.2% and 32.6% respectively). Relationship of distribution of microorganisms belonging to main and additional microflora was revealed. PMID- 19186553 TI - [The role of Paenibacillus polymyxa lectin in the macrophages-mediated killing of bacteria]. AB - Influence of lectin LII of Paenibacillus polymyxa 1460, specific for galactosamine, glucuronic acid, fructose-1,6-diphosphate, and glucosamine, on mechanisms of oxygen-dependent and oxygen-independent killing of bacteria was reviewed. It was shown that the lectin modulates the activity of some microbicide factors in the cytoplasm of macrophages before phagocytosis. Increased activity of myeloperoxidase and acid phosphatase, enhanced formation of active forms of oxygen in alveolar macrophages, increased level of cationic proteins and decreased activity of alkaline phosphatase in peritoneal macrophages was observed. PMID- 19186554 TI - [Multiple antibiotic resistance of associative microflora during urogenital pathology]. AB - Susceptibility of associative microflora isolated from patients with inflammatory diseases of urogenital tract was investigated. Etiologic structure of the diseases and cross-resistance to antibiotics of Escherichia coli, Staphylococcusaureus, and Klebsiella pneumoniae strains isolated from women with endocervicitis and men with urethritiswas assessed. Ciprofloxacin and gentamycin had the highest activity, whereas beta-lactam antibiotics were mildly active. Isolated strainswere resistant to macrolides, penicillines and imipenem. Main types of multidrug resistance to antibiotics were presented in strains circulated in Dagestan. PMID- 19186555 TI - [Growth media for isolation, accumulation, and identification of Listeria in laboratory diagnostics of mix-infections in women with obstetric-gynecological pathology]. AB - Isolation of Lysteria monocytogenes from clinical specimens in obstetric gynecological pathology is often difficult owing to the presence of the mixed microbial flora. Results of study of new Russian growth media for isolation of Listeria for microbiological diagnostics of listeriosis in pregnancy are presented. They show diagnostic efficacy of the developed media for laboratory diagnostics of mix-infections in women with obstetric-gynecological pathology. PMID- 19186556 TI - [Characteristics of Coxsackie A enteroviruses circulating in children in Azerbaijan]. AB - Spectrum of serotypes of Coxsackie A virus and several others markers of virions, which cause enterovirus infection in children in Azerbaijan, was determined. Symptom of myotropism was used as a clinical sign. It was shown that enteroviruses were most frequently detected (41.1%) in children aged registered as enterovirus monoinfection. All isolated strains were characterized by such markers as cytopathic effect, delayed formation of plaques under the layer of agar with low content of sodium bicarbonate. Both attenuated and avirulent virions of Coxsackie A enteroviruses were detected. PMID- 19186557 TI - [Modern taxonomy, classification, and nomenclature of streptococci]. AB - Comparative analysis of main trends of taxonomy, classification, and nomenclature of prokaryotic organisms and streptococci was performed. Phylogenetic classification of streptococci was thoroughly considered. Original classification of streptococci based on pathogenicity and ecological-epidemiological criteria was proposed for medical purposes. Allocation of groups of pathogenic and opportunistic streptococci, which are recommended to place in III and IV groups of pathogenicity, was substantiated. It was shown that according to ecological epidemiological criterion streptococci can be referred to agents of anthroponoses, zoonoses, and sapronoses. PMID- 19186558 TI - [Experimental, clinical and immunologic assessment of acellular staphylococcal vaccine "Staphylovac"]. AB - Results of experimental, clinical and immunological effects of acellular dry staphylococcal vaccine "Staphylovac" developed in Mechnikov Research Institute of Vaccines and Sera are presented. Original mildly virulent strains of Staphylococcus aureus having high immunogenicity, and intra- and interspecies protective activity against different representatives of opportunistic microflora were used for construction of the preparation. Low-toxicity and weak anapylactogenicity of the vaccine were established. In experiments on mice, guinea pigs and rabbits significant protective, antigenic and immunomodulate activity of the preparation was revealed with low sensitization of animals. Clinical trials performed in different centers showed that inclusion of vaccinotherapy in complex treatment of chronic staphylococcal infections (chronic pyodermia, lung abscess etc.) resulted in prolonged pathologic locus, decrease of number and severity of exacerbations, prolongation of remission, and complete recovery in significant number of patients. Activation of innate and adaptive immunity was revealed in the same patients. It was shown on the large group of athletes that administration of the vaccine by aerosol route prevents disruption of immunologic adaptation occurring due to excess physical activity and stress situations during competitions. PMID- 19186559 TI - [Laws of the enteric infections general epidemiology]. PMID- 19186560 TI - Red cell transfusion: how to guide our clinical practice? PMID- 19186561 TI - The first European Antibiotic Awareness Day after a decade of improving outpatient antibiotic use in Belgium. PMID- 19186562 TI - Guidelines for the transfusion of red cells. AB - The following recommendations, which aim at standardising and rationalising clinical indications for the transfusion of red cells in Belgium, were drawn up by a working group of the Superior Health Council. To this end, the Superior Health Council organised an expert meeting devoted to "Guidelines for the transfusion of red cells" in collaboration with the Belgian Hematological Society. The experts discussed the indications for red cell transfusions, the ideal red cell concentrate, the practical issues of administering red cells, and red cell transfusions in patients in a critical condition. The recommendations formulated by the experts were validated by the working group with the purpose of harmonising red cell transfusion in Belgian hospitals. PMID- 19186563 TI - The investigation of a geographical cluster of bladder cancer. AB - Previous analyses of the Limburg Cancer registry (LIKAR) indicated the existence of a geographical cluster of bladder cancer incidence, particularly transitional cell carcinomas, amongst males in the surrounding area of the Belgian cities Hasselt and Alken. In subsequent ecologic analyses no risk factors were identified which could explain the existence of this cluster. Therefore, an epidemiological case-control study has been performed in the province of Limburg to explore the possible determinants of the cluster. The following cluster determinants were investigated: socio-demographic characteristics, life style factors, occupational and environmental exposures, and genetic predisposition. A weight was assigned to all the study subjects. Secondly, we used unconditional logistic regression to calculate crude odd ratios (ORs) and 95% confidence intervals (CIs) for each source of exposure and bladder cancer. Thirdly, we used a multivariate logistic regression analysis which included all the parameters found to be significant at the 0.1 significance level in the univariate analysis. Although some of the specific odds ratios decreased compared with the crude results, the overall excess did not change specifically. In conclusion, we were not able to identify a clear-cut explanation for the existence of the geographical cluster. We therefore recommend an increased surveillance of bladder cancer incidence for 3 to 5 years. If the "excess" of cancer cases has occurred due to random variation, the rate will return to the "expected" range in the next years. If the rate remains elevated, further studies may need to be performed using investigative tools that are not available currently. PMID- 19186564 TI - Hedgehog ringworm in humans and a dog. AB - We report 3 related cases of human dermatophytosis and 1 dog dermatophytosis likely caused by contact with a European hedgehog. Trichophyton erinacei was isolated from stratum corneum samples. This type of zoophilic dermatophytosis is rare in south-east Belgium and probably in the rest of the country as well. PMID- 19186565 TI - Osteosarcoma with extensive calcified pleural metastases at diagnosis. AB - We report the case of an 18 year old woman presenting with shortness of breath and pain in the left shoulder. Imaging of the lungs revealed pleural effusion and calcification of the left pleura. An osteosarcoma of the left humerus was the final diagnosis. A review of the literature reveals that calcified pleural metastatic disease in cases of osteosarcoma has been infrequently reported. Other causes of pleural calcification are briefly discussed. PMID- 19186566 TI - Human bocavirus, a newly discovered parvovirus of the respiratory tract. AB - Human Bocavirus is a newly discovered parvovirus. This virus is the fourth most frequently detected virus among symptomatic children with respiratory infection. Human Bocavirus is present worldwide and is a probable cause of symptomatic respiratory infection, although Koch's postulates are not all fulfilled. In this article, we propose an overview of the main clinical data about this virus, two years after its discovery. In addition, we discuss some hypotheses about its tropism for the lung in young children. PMID- 19186567 TI - Hepatitis C-seroconversion within three to six months after having contracted clinical syphilis and/or lymphogranuloma venereum rectitis in five homosexually active, HIV seropositive men. AB - Five Human Immunodeficiency Virus (HIV) seropositive homosexually active men experienced hepatitis C-seroconversion in the period between September 2004 and January 2007 at a single HIV Reference Center (University Hospital Ghent, Belgium). There was no history of intravenous drug use. All had unprotected anal sex with multiple other HIV seropositive men in the recent past. All of them had clinical syphilis and/or lymphogranuloma venereum rectitis within three to six months before the hepatitis C-seroconversion was detected. This confirms the observations in other case reports and studies originating from the Netherlands, France, the United Kingdom and Germany, illustrating sexual transmission of hepatitis C virus (HCV) infection in this high-risk group. Physicians should be aware of the persistent high-risk behaviour in a subgroup of HIV seropositive homosexually active men and perform intensive sexual counselling and screening for other sexually transmitted diseases, including HCV, during medical follow-up. PMID- 19186568 TI - Donepezil-related toxic hepatitis. AB - Alzheimer's disease (AD) is the leading cause of dementia. It is characterized by the presence of senile plaques and neurofibrillary tangles in the brain, and impairment of the central cholinergic system, which contribute to memory loss and cognitive dysfunction. Cholinesterase inhibitors prevent the hydrolysis of acetylcholine and are currently approved for the symptomatic treatment of Alzheimer's disease. Donepezil, a piperidine-based, reversible and specific inhibitor of acetylcholinesterase, has been demonstrated to be clinically effective in the treatment of patients with mild to moderate AD. To date, clinical trials have not reported an association between treatment with donepezil and hepatotoxicity. We describe a case of toxic hepatitis, documented by liver biopsy, in a patient treated with donepezil. PMID- 19186569 TI - A series of images in localized fibrinolytic treatment for a patient with acute superior mesenteric artery thromboembolism. PMID- 19186570 TI - Lymphangioleiomyomatosis-related spontaneous pneumothorax in a young woman. PMID- 19186571 TI - Breast prosthesis infected by Pasteurella multocida. PMID- 19186572 TI - Waist circumference is associated with asymmetric dimethylarginine levels in patients with metabolic syndrome. PMID- 19186573 TI - Courage and character, leaders and legends: an interview with Joel Cooper, MD, FACS, FRCPS. Interview by Linda Ohler. PMID- 19186574 TI - An opportunity to correct an error and any misimpressions. PMID- 19186575 TI - Skin cancer education in transplant recipients. AB - In the past 20 years, long-term survival for solid-organ transplant recipients has improved dramatically; about 223,000 patients are alive in the United States with organ transplants today. As survival rates improve, however, the morbidity and mortality associated with lifelong immunosuppressive therapy is increasing in significance. Skin cancer is common among recipients of all major organ transplants, including the kidney, liver, heart, lung, and pancreas. Although skin cancer is the most common cancer in transplant recipients, many cases can be prevented by sun protection, skin self-examinations, and physician examinations. Because transplant recipients visit the transplant clinic frequently, clinicians have ample opportunities to teach patients about the importance of prevention and detection of skin cancer. At a routine visit, the clinician should inquire about sun protection practices, especially for tanned, light-skinned, or freckled patients or patients who are planning a warm-weather vacation or time in the sun during the summer. Skin cancer education should be integrated into the care of transplant patients as part of their numerous visits to the transplant clinic. Although some transplant recipients may resist adopting new behaviors at first, use of the ample clinic opportunities for patient education can dramatically reduce their risk of skin cancer. PMID- 19186576 TI - Living kidney donation among Hispanics: a qualitative examination of barriers and opportunities. AB - CONTEXT: Despite their increasing need for kidneys and low nonliving donation rates, minimal research has been conducted to ascertain the perceptions of Hispanic Americans about living organ donation and the process of asking for such a donation. OBJECTIVE: To examine perceptions of Hispanics regarding barriers to and benefits of living donation as well as the process of asking someone to be a living donor. DESIGN: A qualitative study consisting of 10 focus groups conducted in 2 series. PARTICIPANTS: Adult Spanish-language-dominant Hispanic members of the general population of Tucson, Arizona. RESULTS: The main barriers to living organ donation were a lack of knowledge or information and fear of the donation process. Knowing that one has helped save or improve another's life was the central benefit. Most participants reported being willing to ask a relative to be a living donor if they were ever in need. Two main responses typified these individuals: no concern about asking because of a strong desire to fight for one's health and for one's family, or asking despite difficulties and concerns about the process. A significant minority of participants indicated they would not ask for a donation, because of either a desire to avoid harming others or the expectation that a relative would initiate an offer. PMID- 19186578 TI - Communication as a predictor of willingness to donate one's organs: an addition to the Theory of Reasoned Action. AB - Fishbein and Ajzen's theory of reasoned action has been used by many researchers, particularly in regard to health communication, to predict behavioral intentions and behavior. According to that theory, one's intention is the best predictor that one will engage in a behavior, and attitudes and social norms predict behavioral intentions. Other researchers have added different variables to the postulates of attitudes and social norms that Fishbein and Ajzen maintain are the best predictors of behavioral intention. Here we draw on data from a 2006 telephone survey (N = 420) gauging the awareness of an organ donation campaign in Northeast Ohio to examine the impact of communication on people's intentions. The current study supports the hypothesis that those who communicate with others are more likely to express a greater willingness to become an organ donor, but it expands the range of communication contexts. With demographics and attitudes toward organ donation controlled for, this study shows that communication with others about organ donation increases the willingness of individuals to have favorable attitudes about being an organ donor. PMID- 19186577 TI - Improving preemptive transplant education to increase living donation rates: reaching patients earlier in their disease adjustment process. AB - Patients who receive a preemptive kidney transplant before starting dialysis avoid the medical complications related to dialysis and have the highest graft success and lowest mortality rates. Because only 2.5% of incident patients receive kidney transplants preemptively, improved psychosocial education may assist more patients in accessing preemptive transplant. This article outlines (1) unique psychosocial issues affecting patients with chronic kidney disease stage 4 (glomerular filtration rates > 20 mL/min per 1.73 m2) and (2) how an educational program about preemptive living donor transplant should be designed and administered to increase access to this treatment option. Early referral patients may be overwhelmed in coping with and learning about their disease and, therefore, not ready to make a treatment decision, or they may be highly motivated to obtain a transplant to avoid dialysis and return to a normal life. An education program that defines the quality-of-life and health benefits possible with early transplant is outlined. The program is focused on minimizing the disruption of starting 2 treatment techniques and maximizing early transplant health, graft survival, employability, and retention of insurance coverage. Once the benefits of preemptive living donor transplant are outlined, educators can focus on demystifying the living donor evaluation process and assisting interested patients in planning how to find a living donor. To reach all patients, especially racial minorities, education about preemptive transplant should be available in primary-care physicians' and community nephrologists' offices, at dialysis centers, and through other kidney organizations. PMID- 19186579 TI - Short-term impact of an educational program promoting live donor kidney transplantation in dialysis centers. AB - CONTEXT: Given the shortage of kidneys available for transplantation, a community based intervention trial was implemented to assess the impact of an educational program on patients' access to live donor kidney transplantation (LDKT). OBJECTIVE: To compare the short-term impact of a basic intervention and an enhanced intervention on patients' readiness to pursue LDKT. DEGISN: Baseline data and data from 1 week after interventions were analyzed. PARTICIPANTS: 214 transplant-eligible hemodialysis patients attending 14 dialysis facilities in Maryland, Virginia, and Pennsylvania. INTERVENTIONS: In the basic intervention, 107 patients watched a 10-minute videotape on the experience of recipients and live donors of a kidney. In the enhanced intervention, 107 patients watched the same videotape and had a discussion with a health educator on the risks and benefits of LDKT, who could be a donor, and how to address the barriers they were encountering when seeking a live kidney donor. MAIN OUTCOME MEASURE: Patient reported whether they were considering LDKT, had talked with family or friends about LDKT, and had asked someone for a kidney. RESULTS: Over half of transplant eligible patients were not pursuing LDKT at baseline (64% in the basic intervention group, 61% in the enhanced intervention group). One week after the intervention, the odds of considering LDKT were higher among African Americans (odds ratio [OR], 2.28; confidence interval [CI], 1.22-4.25), younger patients (OR, 0.94; CI, 0.91-0.97), and patients who spent less time on dialysis (OR, 0.90; CI, 0.83-0.97). The odds of asking for a kidney were higher among African Americans (OR, 4.94; CI, 2.54-9.60) and patients who perceived they were in poor to fair health (OR, 3.30; CI, 1.12-9.67). CONCLUSIONS: Although both interventions helped patients consider LDKT and ask for a kidney, more time and expanded educational content might be needed to facilitate patients' discussion about LDKT with their loved ones. PMID- 19186580 TI - Design and implementation of an informed consent process before liver transplantation. AB - In the past, the content of patient education was designed at the discretion of each individual transplant center and could vary greatly from center to center. Since the 2007 implementation of the guidelines from the Department of Health and Human Services, Centers for Medicare and Medicaid Services, transplant centers have had to standardize the educational material they give to patients while generating an informed consent form to incorporate this required information. This article describes the development and use of an informed consent form and the educational process for potential liver transplant recipients at Columbia University Medical Center in adherence with the new regulations. This informed consent process can be used as a national model. PMID- 19186581 TI - A multidisciplinary program to educate and advocate for living donors. AB - Education is critical in decision making and the informed consent process in prospective living donors. Little has been written about how and what living donors should be taught. This article describes a multidisciplinary program for living donor education at the University of Virginia. The goals of the program are to impart information needed for prospective donors to make an informed decision and to independently evaluate donors' medical and psychosocial suitability. A partnership between the transplant department and an independent donor advocacy team establishes an environment conducive to education. By embracing independence, transparency, partnership, and advocacy, our program permits bidirectional education. This partnership facilitates unbiased understanding and appreciation of this education and considers each individual's unique circumstances when making informed decisions. Likewise, prospective donors educate the team about their circumstances, which helps the team safeguard the prospective donor and may enhance the safety of prospective donors and the perceived integrity of living organ donation. PMID- 19186582 TI - Designing an educational strategy for increasing organ donor registration among older adults. AB - CONTEXT: Older adults (50-70 years of age) have lower organ donor registration rates than younger adults. Despite recent increases in organ donation registration of adults 18 to 49 years of age, the increase in older adult registrations is less. OBJECTIVE: To use research results from surveys and focus groups to inform an educational strategy and design for the older adult population. DESIGN: Pooled survey data from 3 statewide random-digit dial telephone interview surveys of 6230 adults, including 2631 respondents age 50 or older, were used to identify barriers to organ donor registration and information needs of older adults compared with younger adults. Two focus groups with older adults were used to further identify barriers and to pilot test messages for an educational strategy for older adults. SETTING AND PARTICIPANTS: Surveys were conducted with adults in Ohio. Focus groups were conducted in 2 major cities in Ohio with older adults, 50 to 70 years of age. RESULTS: Survey results indicate that older adults and younger adults differ on several factors regarding their beliefs, attitudes, and behavior concerning organ donation. Although older adults express positive attitudes toward donation, they often "opt out" from registering to be organ donors for age and health reasons. Focus groups show similar barriers to organ donor registration and indicate that brochures are a preferred way to educate older adults about organ donation. CONCLUSION: Using research results to identify barriers and information needs of specific demographic groups is very important in designing a targeted educational strategy. An educational campaign addressing the concerns and information needs of this age group could increase organ donor registration. PMID- 19186583 TI - Adapting teaching styles to accommodate learning preferences for effective hospital development. AB - Hospital development professionals in organ procurement organizations engage in many roles in the hospital. The important role of educator requires making the most of each teaching opportunity by understanding the characteristics of the learning audience and applying proven principles of adult educational design, with a focus on collaborative learning and variety in presentation techniques. The goal is to provide effective education, enabling hospital staff to transfer this learning to the job setting, with the outcome of facilitating a supportive process that saves lives through organ donation. PMID- 19186584 TI - Effects of information, education, and communication training about organ donation for gatekeepers: clerks at the Department of Motor Vehicles and organ donor registries. AB - CONTEXT: Clerks at the Department of Motor Vehicles (DMV) serve as gatekeepers in the quest to fill organ donor registries. Most people who join organ donor registries do so through their local driver's license bureau. OBJECTIVE: To improve knowledge, attitudes, and behavior toward organ donation among DMV clerks, and to improve strategies for communicating with the public about organ donation, resulting in more people joining DMV-based organ donor registries. SETTING: DMV offices in 8 counties in a southern state. PARTICIPANTS: A general survey of 1504 participants in 8 counties was conducted to assess knowledge, attitudes, beliefs, and behaviors toward organ donation. All 76 DMV clerks in those counties participated in surveys before and after a training intervention. INTERVENTIONS: DMV clerks received a 1-hour training intervention focused on the nature of the new organ donor registry, information about organ donation, and communication strategies for interacting with the public. MAIN OUTCOME MEASURES: Knowledge, attitudes, beliefs, behaviors, and behavioral intentions toward organ donation were measured for the general public survey and for DMV clerks. RESULTS: Clerks were more knowledgeable about brain death and religious stances on organ donation than the general public, but otherwise both groups shared similar knowledge, attitudes, beliefs, behaviors, and behavioral intentions toward organ donation. Overall the general public and clerks had favorable attitudes toward organ donation but low to moderate knowledge, beliefs, and behaviors and behavioral intentions. Training significantly increased DMV clerks' knowledge, attitude, beliefs, and behavioral intention toward donation. Organ donor registration rates were a mean of 14% higher in counties where clerks were trained than in control counties and were 9% higher than statewide. CONCLUSIONS: Training DMV clerks is an effective way to increase knowledge, attitudes, and beliefs for gatekeepers of organ donor registries and may increase donor registration rates among the public. PMID- 19186585 TI - Organ tourism. PMID- 19186586 TI - If not now then when? PMID- 19186587 TI - Middlemen. PMID- 19186588 TI - No physician bailout. PMID- 19186589 TI - Maryland medicine. Introduction. PMID- 19186590 TI - Ending homelessness with Maryland's Health Care for the Homeless, Inc. PMID- 19186591 TI - A big step forward for Maryland on health care: affordable health care for all Marylanders. PMID- 19186592 TI - Housing First: a prescription for success. PMID- 19186593 TI - Homelessness and our most vulnerable patients. AB - The scope of homelessness among children is broad and growing, and its affect on physical and mental health is extensive. It may seem daunting for individual providers to make an impact on the challenges faced by these most vulnerable of patients. However, healthcare providers who care for homeless children can improve more than just their physical health by understanding barriers specific to this population, and addressing them through minor changes in standard practice; education of self, staff, and colleagues; and advocacy. By collaborating with parents and local agencies, clinicians can make tangible progress in improving the health of their homeless patients and can provide parents with the information and support they need to prioritize a child's health needs appropriately. Ultimately, providers should strive to make their practices a true medical home, as it may be the only home a child knows. PMID- 19186594 TI - Infectious disease in the homeless. AB - Homelessness increases a person's exposure to infectious and communicable diseases. The barriers to treatment as described above are many for the physicians and other providers of health care to this population. Health Care for the Homeless would greatly welcome the help of medical subspecialists in the areas of infectious disease, dermatology, gastroenterology, and pulmonology for some of our particularly challenging patients. We look forward to continuing our collaborations with the medical community in Baltimore City as we serve its most vulnerable citizens. PMID- 19186595 TI - Improving access to treatment for patients with addiction: a physician's reflections on addiction treatment. PMID- 19186596 TI - Mental illness in homeless people. PMID- 19186597 TI - The homeless veteran: call to action and response. PMID- 19186598 TI - Homelessness and early death. PMID- 19186599 TI - Health Care for the Homeless, Inc. Personal stories from former clients. PMID- 19186600 TI - No medicine is good medicine. PMID- 19186601 TI - The last word. PMID- 19186602 TI - Improving mortality from stroke disease: putting the evidence into practice. PMID- 19186603 TI - A breath of fresh air: a new UK guideline for emergency oxygen therapy. PMID- 19186604 TI - Stents and stentability: treatment for malignant bowel obstruction. AB - Colonic stents offer a palliative treatment for patients with malignant bowel obstruction otherwise requiring surgery and possible stoma, or as a bridge to surgery for potentially curative malignant disease. This article reviews the indications, risks and benefits of stent insertion. PMID- 19186605 TI - Recent advances in the management of rectal cancer. AB - Recently there have been significant advances in the management of patients with rectal cancer, attributed mainly to advances in surgical techniques and pre- and postoperative therapy. This review addresses some of these advances and their impact on the prognosis for rectal cancer patients. PMID- 19186606 TI - Acute mesenteric ischaemia: facts and perspectives. AB - Acute mesenteric ischaemia is a catastrophic abdominal emergency with an extremely high mortality rate. This article discusses the aetiology, diagnosis and treatment of acute mesenteric ischaemia with emphasis on avoidance of common errors that contribute to the poor outcome inherent to this condition. PMID- 19186607 TI - Cardiac device therapy 2: indications, techniques and complications. AB - With advancing technology and ever-expanding indications for implantable cardiac pacing and defibrillation devices, this article reviews modern day practice in this field. This article focuses on topics pertinent not only to cardiologists but also to general physicians, medical trainees and allied medical specialties. PMID- 19186608 TI - The survivors' guide to working in the medical admission unit. PMID- 19186609 TI - NHS jargon buster. PMID- 19186610 TI - Making the most of the doctors' mess: a guide for the foundation year doctor. PMID- 19186611 TI - A guide to assisting at surgical operations 1: general principles. PMID- 19186612 TI - Addison's disease: a diagnostic challenge. PMID- 19186613 TI - So you want to be...a paediatrician. PMID- 19186614 TI - Management of tuberculosis. PMID- 19186615 TI - Richard Bright (1789-1858): pioneer of nephrology. PMID- 19186616 TI - National on-line recruitment to foundation programmes in the UK: evolution and evaluation. PMID- 19186617 TI - Revascularization of the amputated limb in symptomatic arteriopaths: an underused treatment. PMID- 19186618 TI - Spontaneous lower limb subcutaneous emphysema: a diagnostic dilemma. PMID- 19186619 TI - Massive pneumoperitoneum in an infant: the football sign. PMID- 19186620 TI - An unusual case of jaundice. PMID- 19186621 TI - Cardiac resynchronization therapy in chronic heart failure with atrial fibrillation. PMID- 19186622 TI - Resolve to brush up your finances in 2009. PMID- 19186623 TI - Sugammadex: will it change anaesthetic practice? PMID- 19186625 TI - This year's challenge: maintaining quality care under a tight budget. Interview by Tamsin Snow. PMID- 19186626 TI - Nurses in Florence Nightingale's day worked 'until they dropped'. PMID- 19186628 TI - Care for the whole city. AB - A report criticising cancer care in the capital led to the development of Action for London, a project that involves nurses working out how to put their vision for a better service into practice. PMID- 19186627 TI - Meet the next generation. Interview by Alison Whyte. AB - Nursing Standard profiles five nursing students who embarked on their studies at the University of Liverpool in September 2008. Readers will be able to find out about their experiences half way through the course and again at the end. PMID- 19186629 TI - Identifying impetigo. Interview by Lynne Pearce. AB - Impetigo is a common condition, particularly among children. But it is often misdiagnosed. PMID- 19186630 TI - Recession reality bites. PMID- 19186631 TI - Avoiding plagiarism in academic writing. AB - Plagiarism means taking the work of another and presenting it as one's own, resulting in potential upset for the original author and disrepute for the professions involved. This article aims to explore the issue of plagiarism and some mechanisms for detection and avoidance. PMID- 19186632 TI - Interpretation of competence in student assessment. AB - The Nursing and Midwifery Council's (2008a) Standards to Support Learning and Assessment in Practice have been strengthened, with an expectation that mentors develop their knowledge and skills beyond registration to enable credible decisions about the competence of students. As a result, there are a number of conflicts for students and mentors to negotiate, not least in maintaining a fair and transparent assessment process. This review identifies different interpretations of competence and competence assessment. Competence assessment should be given greater credibility when student learning is set in a context of care involving patient experience and evaluated by reflexive and inclusive mentors. PMID- 19186633 TI - Assessment and management of patients with hypothyroidism. AB - This article explains how to assess, diagnose and treat patients with an underactive thyroid gland (hypothyroidism), including an outline of replacement therapy options. The need for regular monitoring and ongoing support is also highlighted. PMID- 19186634 TI - Psychological support. PMID- 19186635 TI - First interview success. PMID- 19186636 TI - Retirement can be a new beginning. Interview by Lynne Wallis. PMID- 19186637 TI - Collaboration, not combat. PMID- 19186638 TI - Note for tough times: don't rob St. Paul to pay St. Peter. PMID- 19186640 TI - Fostering an ethical culture: rules are not enough. PMID- 19186639 TI - California hospital, school district collaborate to cut obesity-related risks. PMID- 19186641 TI - Ownership defined differently in civil, canon law. PMID- 19186642 TI - Reform now? Reform how? PMID- 19186643 TI - Yes! The time is now. Health care reform can help the country's economic recovery process. PMID- 19186644 TI - 'A new day in Washington'. Catholic health care leader sees rare opportunity ahead for health care reform. PMID- 19186645 TI - Bound by fear: health care reform means letting go. The public's resistance to change clashes with responsibilities to justice and the common good. PMID- 19186646 TI - The European experience: basis for U.S. reform? Many countries offer models, but everywhere the key is controlling costs. PMID- 19186647 TI - Catholic values and the market: friends or foes? Can Catholic health care support market-based reforms? PMID- 19186648 TI - Doing better with what we have--key to successful reform. Waste is silver lining in black economic cloud. PMID- 19186649 TI - Achieving high performance preventive care. The Commonwealth Fund's five strategies aim to realize comprehensive performance improvement. PMID- 19186650 TI - Long-term care financing reform: the time is here. Needed: proposals that promote accessibility and affordability. PMID- 19186651 TI - Beyond reform: Catholic health care matters even when all can pay. Access is just one dimension of the ministry's distinctive mission. PMID- 19186652 TI - 'Do no harm': time to turn a phrase on its head. PMID- 19186653 TI - Mental health benefits get boost from federal law. Hospital leader discusses impact of new legislation. PMID- 19186654 TI - Maria Del Rio-Hoover, MD. A tireless advocate for children. PMID- 19186655 TI - Too much of a good thing but worth the work. PMID- 19186656 TI - Employee embezzlement: autonomy provides opportunity for an office manager to take advantage of her position. PMID- 19186657 TI - Dental occlusion and periodontal disease: what is the real relationship? AB - ABSTRACT The role of occlusion in periodontal disease has always been a challenging topic. A good understanding of the current status of the relationship of occlusion and periodontitis is of paramount importance in order for dental clinicians to provide adequate and comprehensive periodontal treatment in patients presenting with traumatic occlusion. This article reviews the literature regarding the relationship between occlusion and periodontitis and presents recommendations for clinical practice based on available evidence. Clinical cases illustrating the complexity of this relationship and their management are presented. PMID- 19186658 TI - Increasing dental care for very young children: what can training accomplish? AB - Too few dental providers feel comfortable or are keen on seeing young children in their practices, and training in oral health has generally ignored the dental component of early childhood. Evaluation of California's $7 million First Smiles showed increased knowledge and skills among 3,369 dental professionals trained. Positive practice changes included increasing willingness to see more 1-5 aged children, including special needs; seeing children for a first visit by age 1; and conferring with a pregnant patient's medical provider. PMID- 19186659 TI - Use of prophylactic antibiotic therapy in oral surgical procedures: a critical review. AB - Despite myriad evidence-based data on the efficacy of prophylactic antibiotics in oral surgical procedures, their inaccurate use and indiscriminate abuse still continues in all clinical settings. Although controversy remains, clear-cut guidelines relying on established scientific principles do exist and must be followed. This paper provides a critical and systematic review of each principle and encourages practitioners fit to what current evidence demands. PMID- 19186661 TI - Research: something to sink your teeth into. PMID- 19186660 TI - Effect of preflaring on working length determination: an in vivo study. AB - This study is designed as an in vivo study to evaluate the impression of the preflaring method on working length determination in curved canals. PMID- 19186662 TI - Silent PPOs spur doctor push-back. PMID- 19186663 TI - Generic drug price war begins to benefit insurers. PMID- 19186664 TI - Specialists warming to pay for performance. PMID- 19186665 TI - Disruptive innovations that will change your life in health care. PMID- 19186666 TI - Network-model health plans may be tomorrow's dinosaurs. Interview by John Marcille. PMID- 19186667 TI - Health plans: disrupt and prosper. PMID- 19186668 TI - Don't get trapped by PBMs' rebate labeling games. PMID- 19186669 TI - Multifaceted diabetes program pays off for healthpartners. PMID- 19186670 TI - E-prescribing system increases generic usage. PMID- 19186671 TI - Empowered physicians are key to diabetes program's success. PMID- 19186673 TI - 2009 costs will continue to outpace inflation. PMID- 19186672 TI - Orphan disease gains second treatment option. PMID- 19186674 TI - Help patients, families cope with end-of-life situations. PMID- 19186675 TI - Get in touch with your feelings about EOL issues. PMID- 19186676 TI - Throughput measures decrease LOS in ED. PMID- 19186677 TI - Multidisciplinary rounds at bedside involve patients, families. PMID- 19186678 TI - Program targets pneumonia quality measures. PMID- 19186679 TI - Make sure patients are appropriate for home care. PMID- 19186680 TI - [Asthma--between "control" and "severity"]. PMID- 19186681 TI - [Multidrug resistant tuberculosis in Romania in the last years (2004-2007)--an extremely important social phenomenon]. AB - The emergence of resistance to drugs used to treat tuberculosis (TB), and particularly multidrug resistant (MDR-TB) strains, has become a significant public health problem in a number of countries and an obstacle to effective global TB control. In many other countries, the extent of drug resistance is unknown and the management of the patients with MDR-TB is inadequate. In countries where drug resistance has been identified, specific measures need to be taken within TB control programmes to address the problem through appropriate management of patients and adoption of strategies to prevent the propagation and dissemination of drug resistant TB, including MDR-TB. In Romania there are two centers for the management of MDR-TB cases: one in Bucharest at The National Pneumology Institute "Marius Nasta" and the other at Bisericani (Neamt district). The comparative analysis of MDR-TB cases and under treatment evolution was done, in the two excellence centers Bucharest and Bisericani from 2004 to 2007. This is a retrospective study of MDR-TB patients enrolled in Romania between 2004-2007. All patients evaluated were managed under The National Tuberculosis Programme approved protocols and had the opportunity to receive more than 24 months of treatment. In addition, follow-up data on successfully treated patients were collected at the beginning of 2007. This study is based on an MDR register, and a software collecting information on MDR-TB cases. In order to be accepted in one of the two MDR centers, patients need to fulfill certain criteria to improve the treatment results. A total number of 305 MDR-TB patients were registered at the beginning of 2007 in Bucharest MDR center; this study used a number of 170 MDR-TB patients from Bucharest who fulfilled the study's criteria in oppose to the 294 MDR-TB patients from Bisericani. 184 patients from the first cohort of DOTS-Plus project were evaluated until the end of the study: 55 were completed treatment and 54 cured, which means 59,23% succes rate. The conversion rate of culture at 4 months (for the first cohort) in MDR excellence centers was: Bucharest--72,5% and Bisericani--64%. PMID- 19186682 TI - [Pre-neoplastic bronchial lesions: possibilities of diagnosis and chemo prevention]. AB - Early diagnosis of pulmonary cancer is crucial in the fight against this disease. Pre-neoplastic bronchial lesions are the first stage in the evolution towards invasive cancers. This paper describes histological patterns of these pre neoplastic bronchial lesions, as well as early, intermediate and late molecular disturbances associated with this condition. The ways to early identification of these lesions are: a) bronchoscopy with laser-induced auto-fluorescence guided through Optical Coherence Tomography, which is able to identify cellular and extracellular structures, from the cellular surface or beneath it; b) PET-CT which identifies lesions over 6 mm; c) bio-markers: the sensitivity of the test is 60-70%; the positive predictive value is higher when these markers are taken together. p53 and p16-INK4A genes seem to have the higher predictive value. Identification of these disturbances leads to improved possibilities of chemo prevention. PMID- 19186683 TI - [New data in staging and prognosis of bronchopulmonary carcinoma. Part II: volumetric analysis of pulmonary nodules]. AB - Pulmonary nodules discovered incidentally or in the context of the work-up for symptomatic conditions represent an area of major interest. By definition, a pulmonary nodule is defined as a well-circumscribed round or oval lesion, measuring less than 3 cm in maximal diameter. The impact of newer technologies on our capacity to detect pulmonary nodules has increased significantly in the last decade, from the utilization of high-performance computed tomographic (CT) scanners, to the development of more functional imaging techniques, such positron emission tomography (PET) scanners. The latter still has a suboptimal resolution for nodules less than one centimeter in size, hence the use of high-resolution, multi-detector CT scanners has become a more frequent clinical problem for these nodules. In this review we describe the latest developments in the CT technology, such as volumetric reconstruction and characterization of the pulmonary nodules and how this can impact the modern diagnostic and therapeutic modalities. PMID- 19186684 TI - [Prevalence of sleep apnoea syndrome--the experience of "Marius Nasta" Institute of Pneumology, Bucharest]. AB - The study is a retrospective one based on the analysis of the records of the patients who have been diagnosed with Sleep Apnoea Syndrome (SAS) in "Marius Nasta" Institute of Pneumology sleep laboratory between 1997-2007. The study demonstrated a SAS prevalence of 64,4% in a specific population. Snoring is the main symptom which brings the patients to the doctor (74,5%); regarding the severity of the disease 62,4% were moderate and severe cases. CONCLUSIONS: Sleep Apnoea Syndrome is a frequent disease, relatively easy to diagnose and to treat. If we want to have real epidemiological data regarding SAS prevalence in Romania we should have large epidemiological studies in all the regions of the country. PMID- 19186686 TI - [Pulmonary epithelioid hemangioendothelioma--a case report]. AB - Pulmonary epithelioid hemangioendothelioma (PEH) is a rare soft tissue tumor of endothelial origin that occurs among young women and typically presents as bilateral multiple nodules, readily mistaken for carcinoma or, as in this case, Wegener's granulomatosis. This is a rare disease, with approximately 50 cases described in the literature. In the present report, we describe a case of PEH in a 39-yr-old woman. Immunohistochemically, the tumor cells were positive for CD34. PMID- 19186685 TI - [Particularities of the association of systemic arterial hypertension in patients with sleep apnea syndrome]. AB - AIM: Analysis of the association between sleep apnea syndrome (SAS) and systemic hypertension (SHT) in consecutive patients with suspicion of SAS. METHOD: We included 427 consecutive patients evaluated for SAS at Babes University Sleep Lab, Timisoara (March 2001-March 2007). Medical history, anthropometric measurements, cardiovascular evaluation, inclusion/exclusion criteria, Epworth scale, polysomnography, statistical analysis with STATA 9.2. were done. RESULTS: Systemic hypertension (SHT) at 245 (58,05%) of patients, pre-SHT at 11,37%, stage I 11,37%, stage II 29,14%, stage III 6,16%, onset of disease 7,26 +/- 6,13 years. Differences between groups without and with SHT: apnea-hypopnea index (AHI) 27,71 +/- 27,94/h, CI 23,55-31,78 vs 38,6 +/- 26,16/h, CI 35,34-41,85, p < 0,001, body mass index (BMI) 31,43 +/- 6,26 kg/m2, CI 30,50-32,36 vs 34,21 +/- 6,11 kg/m2, CI 33,45- 34,97 p < 0,001, desaturation index 20,21 +/- 26,35/h, CI 16,28-24,13 vs 31,12 +/- 29,16 CI 27,49-34,75, p < 0,001, medium desaturation 92,93 +/- 5,03%, CI 92,18-93,68 vs. 90,6 +/- 8,42, CI 89,55-91,64, p < 0,001, medium age 47,66 +/- 12,57 years, CI 45,79-49,52 vs. 55,06 +/- 10,35 years, CI 53,77-56,35, p = 0,0047, abdominal circumference 117,77 +/- 15,44 cm, CI 115,48-120,06 vs. 121,68 +/- 14,66 cm, CI 119,85-123,51, p = 0,0082, neck circumference 42,98 +/- 4,84 cm, CI 42,26-43,70 vs. 44,24 +/- 4.59 cm, CI 43,67-44,81 p = 0,0068, Epworth score 9,09 +/- 4,81, CI 8,37-9,80 vs. 10,77 +/- 5,19, CI 10,12-11,42, p = 0,007. CONCLUSION: SHT represents the most frequent disease association in SAS patients. Differences between groups without and with SHT were statistical significant in apnea--hypopnea index, body mass index, desaturation index, medium desaturation, age, abdominal circumference, neck circumference, Epworth score. PMID- 19186688 TI - [Hyperthermic intrapleural chemotherapy--another modality for the treatment of malignant pleural neoplasia]. AB - The high frequency and the severity of malignant pleurisies, despite their etiology, make the problem to be of great importance. At national level, the problem of neoplastic pleurisies was and still remains a medico-surgical problem with socio-economic implications, especially in the situation when cancer represents a major health problem for the national health system. Treatments are mostly palliative aiming to relieve dyspnea and thoracic pain. The increasing incidence of malignant pleurisies enforce a better knowledge of pathogenic mechanisms that led to a new therapeutic approach with promising results- cytoreductive surgery combined with intraoperative hyperthermic intrathoracic chemotherapy. Recent studies about compared results of different types of treatments, function of survival period and side-effects, demonstrate the advantages of association between surgical treatment with the intrapleural administration of hyperthermic chemotherapy, which may have control on pleurisy and pleural involvement. It is considered that hyperthermia represents a hope in increasing the quality of patient's life with radiotherapy and chemotherapy, as well as surgical treatment, resistant neoplastic pleurisy. It is necessary to be made in accordance with classical oncological treatments. PMID- 19186689 TI - [Polonium: the radioactive killer from tobacco smoke]. AB - Among all carcinogenic substances contained in tobacco smoke, Polonium 210 (Po 210), with a half-life of 138 days, is one of the most dangerous, by exerting a devastating, chronic, slow and progressive carcinogenesis activity. The main source of Po-210 in tobacco is represented by fertilizers (polyphosphates) containing radium-226 (Ra-222) which decades to plumb 210 (Pb-210). Through the thricomes Pb-210 is concentrated in the tobacco leaves, where it turns to Po-210, which at the cigarette combustion temperature (800-900 degrees C) reaches the gaseous state and it is absorbed by the micro particles released into tobacco smoke. Thus, smoke becomes radioactive in both its gaseous and corpuscular components and reaches the airways, where, particularly at the branches level and together with other substances, it exerts its carcinogenic activity, especially in those subjects with impaired respiratory mucosal clearance. The carcinogenic risk/one year lifetime of a smoker of 20 cigarettes per day is equivalent to that of undertaking 300 chest x-rays. It is calculated that Po-210 may be independently responsible of 4 lung cancers every 10,000 smokers. During cigarette's combustion, tobacco smoke is also released in the air, contributing to serious health risks for those exposed to passive smoke. PMID- 19186690 TI - [Romanian Symposium of Pneumology, October 23-24, 2008, Satu Mare]. PMID- 19186691 TI - [Romanian Society of Pneumology included in GARD (Global Alliance Against Chronic Respiratory Diseases)]. PMID- 19186692 TI - [The Second National Conference on Physiopathology of Respiratory System, Brasov, Romania, October 31-November-1, 2008]. PMID- 19186694 TI - Spreading the health. PMID- 19186693 TI - [A small Congress can be helpful. The 10th International Congress of Research on Tobacco and Nicotine, Roma, September 23-26, 2008]. PMID- 19186695 TI - Detours by design. PMID- 19186697 TI - Quantum afterlife. PMID- 19186696 TI - Plague in the prairie. PMID- 19186698 TI - Shell game. PMID- 19186699 TI - Childhood recovered. PMID- 19186700 TI - Acid bath. PMID- 19186701 TI - A scientific stimulus for the U.S. PMID- 19186702 TI - Transforming the auto industry. PMID- 19186703 TI - Darwin misunderstood. PMID- 19186704 TI - Naked singularities. PMID- 19186705 TI - Nanomedicine targets cancer. PMID- 19186706 TI - The origin of the land under the sea. PMID- 19186707 TI - New dawn for electric rockets. PMID- 19186708 TI - Sculpting the brain. PMID- 19186709 TI - The greenhouse hamburger. PMID- 19186711 TI - Touch screens redefine the market. PMID- 19186710 TI - Chaos and the catch of the day. PMID- 19186712 TI - [Therapeutic effect of acupuncture on cisplatin-induced nausea and vomiting]. AB - OBJECTIVE: To observe therapeutic effect of acupuncture combined with antiemetic on cisplatin-induced nausea and vomiting. METHODS: By using paired, cross controlled trial design, 66 cases of chemotherapy were divided into group A and B, 33 cases in each group. For the group A, chemotherapy, tropisetron and acupuncture therapy were adopted in the first chemotherapy cycle and the same chemotherapy program, tropisetron and sham acupuncture were used in the next cycle. For the group B, chemotherapy, tropisetron and sham acupuncture were given in the first chemotherapy cycle and the same chemotherapy program, tropisetron and acupuncture therapy were applied in the next cycle. Zusanli (ST 36), Neiguan (PC 6) and Gongsun (SP 4) and auricular point Wei (stomach) were selected for acupuncture therapy, and the points at 3 cm lateral to Zusanli (ST 36) , Neiguan (PC 6) and Gongaun (SP 4) and auricular point corresponding to scapha level were selected for sham acupuncture. Acupuncture treatment or sham-acupuncture was given for 6 consecutive days, once each day and antiemetic tropisetron 5 mg was given to the two groups as basic antiemetic prophylaxis for 6 days, once daily. The therapeutic effects on nausea and vomiting in the 6 days were compared between the acupuncture group and the sham-acupuncture group in the two chemotherapeutic cycles. RESULTS: The effective rates for nausea in the 2nd day and the 4th day were 87.1% and 79.0% in acupuncture group, which were superior to 59.4% and 57.8% in the sham-acupuncture group, respectively (both P < 0.05); and the therapeutic effects on vomiting in the 3rd-6th day in the acupuncture group were better than those in the sham-acupuncture group (P < 0.05). CONCLUSION: Acupuncture combined with antiemetic can effectively decrease the incidence and degree of cisplatin-induced delayed nausea and vomiting. The effect of acupuncture is better than that of sham acupuncture. PMID- 19186713 TI - [Influence of different frequencies of acupuncture on therapeutic effect in patients with cerebral infarction at convalescence]. AB - OBJECTIVE: To observe the influence of different frequencies of acupuncture on the therapeutic effect in patients with cerebral infarction at convalescence. METHODS: Ninety-seven cases were randomly divided into an observation group I (n = 50) and an observation group II (n = 47). They were treated with same Chinese drugs and western medicine and electroacupuncture at Jiquan (HT 1), Quchi (LI 11), Hegu (LI 4), Huantiao (GB 30), etc. The observation group I was treated twice each day and the observation group II once each day. After treatment of 30 days, their therapeutic effects were observed. RESULTS: The total effective rate of 94.0% for improvement of limb activity in the observation group I was better than 78.7% in the observation group II (P < 0.05); the therapeutic effects for choking when taking water, dysphagia, vague mind and slurred speech were similar in the two groups (P > 0.05). CONCLUSION: The therapeutic effect of acupuncture twice each day on cerebral infarction at convalescence is superior to that of once daily. PMID- 19186714 TI - [Influence of garlic moxibustion on the therapeutic effect in re-treatment patients of tuberculosis]. AB - OBJECTIVE: To observe the therapeutic effect of moxibustion in re-treatments patients of tuberculosis. METHODS: Fifty-three cases were randomly divided into an observation group (n = 31) and a control group (n = 22). They were treated with routine chemotherapeutic program of western medicine with garlic moxibustion on main points Feishu (BL 13), Gaohuang (BL 43), Shenzhu (GV 12), etc. added in the observation group. The therapeutic effects were assessed by clinical symptoms and signs, X-ray, CT examination and laboratory indexes. RESULTS: The focus absorbing rate of 87.1% in the observation group was better than 63.6% in the control group (P < 0.05); the rate of bacteria-turned negativity in sputum was 90.5% in the observation group which was better than 56.3% in the control group (P < 0.05); the observation group in improvement of hypodynamia, night sweat and cough was superior to the control group (all P < 0.05). CONCLUSION: Moxibustion can increase the therapeutic effect for the re-treatment patient of tuberculosis. PMID- 19186715 TI - [Control study on acupuncture and medication for treatment of primary simple premature ejaculation]. AB - OBJECTIVE: To compare therapeutic effects of acupuncture and medication on primary simple premature ejaculation. METHODS: One hundred and eleven cases were randomly divided into an acupuncture group (n = 56) and a medication group (n = 55). The acupuncture group was treated by acupuncture with acupoint group I Xinshu (BL 15), Ganshu (BL 18), Pishu (BL 20), Shenshu (BL 23) and acupoint group II Guanyuan (CV 4), Zhongji (CV 3), Sanyinjiao (SP 6), Taixi (KI 3), Taichong (LR 3) alternately selected, one acupoint group daily. The medication group was treated with oral administration of Sailete tablets, 20 mg each tablet, one tablet each night. After they were treated for one course, their therapeutic effects were compared. RESULTS: The total effective rate was 82.1% in the acupuncture group and 63.6% in the medication group; the cumulative score after treatment was 12.56 +/- 3.84 in the acupuncture group and 11.50 +/- 3.77 in the medication group, with a significant difference between the two groups in the therapeutic effect (P < 0.05). CONCLUSION: Acupuncture has a better therapeutic effect on primary simple premature ejaculation. PMID- 19186716 TI - [Clinical study on acupuncture combined with moxibustion on temperature-sensitive points for treatment of Bell palsy in the acute stage]. AB - OBJECTIVE: To probe into a new therapy with less pain, good therapeutic effect and convenience for facial palsy in acute stage. METHODS: Sixty cases were randomly divided into 2 groups, an observation group and a control group, 30 cases in each group. The observation group was treated with routine acupuncture plus moxibustion on temperature-sensitive points, and the control group with the routine acupuncture. Their therapeutic effects were assessed by the criteria for assessment of facial nerve function stipulated by Japanese Institute for Researching the Facial Nerve in 1997. RESULTS: There were significant differences in cumulative scores of symptoms before and after treatment in the two groups (both P < 0.01) and there was a significant difference in the cumulative score of symptoms after treatment between the two groups (P < 0.01). The cured rate was 76.67% in the observation group and 50.00% in the control group with a significant difference between the two groups (P < 0.05). The total effective rates were respectively 93.33% and 76.67% in the two groups with no significant difference between the two groups (P > 0.05). CONCLUSION: Both acupuncture combined with moxibustion on temperature-sensitive points and simple acupuncture have good therapeutic effects on idiopathic facial palsy, but the observation group is better than the control group. PMID- 19186717 TI - [Clinical observation on acupuncture combined with medication for treatment of continuing unovulation infertility]. AB - OBJECTIVE: To explore the therapeutic effect of acupuncture combined with medication on continuing unovulation infertility. METHODS: Fifty cases of continuing unovulation infertility were randomly divided into an observation group and a control group, 25 cases in each group. The observation group was treated with Chinese herbal decoction plus acupuncture. Culuanpao decoction, Cupailuan decoction and Cuhuangti decoction were respectively given according to different menstrual cycles and acupuncture and moxibustion at Guanyuan (CV 4), Zhongji (CV 3), Luanchao (ovary), etc. The control group was treated with oral administration of Clomiphene and intramuscular injection of Chorionic Gonadotropin. They were treated for 6 cycles and the ovulation rate and the pregnancy rate were observed. RESULTS: There was no significant difference in the ovulation rate between the two groups (P > 0.05). The pregnancy rate of 44.0% in the observation group was significantly higher than 16.0% in the control group (P < 0.05). The observation group was superior to the control group in the score of mucus and the endometrial thickness. CONCLUSION: Both acupuncture combined with medication and oral administration of clomiphene plus intramuscular injection of chorionic gonadotropin have higher ovulation rate, and the former was higher than the latter in the pregnancy rate. PMID- 19186718 TI - [Analgesic effect of acupuncture compound anesthesia for patients of different pain thresholds]. AB - OBJECTIVE: To observe analgesic effect of acupuncture compound anesthesia for patients of different pain thresholds. METHODS: Seven hundred cases of infertility who will be taken ova under ultrasound guidance via the va gina and receive in vitro fertilization and embryo transfer (IVF-ET), were randomly divided into an acupuncture compound anesthesia group (n = 347) and a simple Dolantin group (n = 353). The pain thresholds in the acupunc ture compound anesthesia group was determined 30 min before acupuncture on one day before taking ova, and in the simple Dolantin group was determined before injection of Dolantin and 30 min before taking the ova. The patients in the two groups respectively received electroacupuncture at Shenshu (BL 23), Ciliao (BL 32), Baihui (GV 20), Sanyinjiao (SP 6), etc. combined with intramuscular injection of Dolantin, and simple intramuscular injection of Do lantin. RESULTS: For the patients with pain thresholds of 1 mA, 1.5 mA and 2 mA, the cumulative scores of pain in the acupuncture compound anesthesia group were significantly lower than those in the simple dolantin group (P < 0.01, P < 0.05). For the patients with pain thresholds of 0.5 mA, 2.5 mA and 3 mA, in cumulative scores of pain there were no significant differences between the two groups (P > 0.05). CONCLUSION: For the patients with moderate pain thresholds, acupuncture compound anesthesia has good analgesic effect. PMID- 19186719 TI - [Inflammatory reaction caused by acupuncture is one of the initial factors of acupuncture effect]. AB - The modern studies indicate that there is a close relationship between mast cells and the acupuncture effect, and acupuncture can activate mast cells to induce a series of vascular reaction and immunological effect. The authors hold that acupuncture is a kind of nociceptive stimulus, which can cause inflammatory reaction in the sites of acupuncture, and then further activate the nerve endocrine-immune network to cause the cascade amplification of the acupuncture effect. The inflammatory reaction induced by acupuncture is one of the initial factors of acupuncture effect. PMID- 19186720 TI - [Remarks on application of acupuncture instruments in acupuncture quantification and normalization studies]. AB - Acupuncture manipulation quantification is an important link of acupuncture normalization study. Because traditional acupuncture manipulation are complicated with difficulty of quantification, acupuncture instruments provide a new way for acupuncture manipulation quantification and acupuncture normalization. It is necessary to increase the understanding of the importance of developing acupuncture instruments, strengthen the study of related theory and development of acupuncture measurement instruments and acupuncture imitation instruments, enlarge serviceable range, verify efficacy, develop the theory of acupuncture manipulation, richen the study methods of acupuncture normalization, so as to promote internationalization of acupuncture. PMID- 19186721 TI - [Thinking about standard and standardization of acupuncture and moxibustion]. AB - Emphasis on standardization development of acupuncture-moxibustion science is very important. In ancient times, the theory of Chinese acupuncture and moxibustion was in agreement consistent. Illustrated Manual of Points for Acupuncture and Moxibustion on a Bronze Statue with Acupoints (A. D. 1026) and bronze statue with acupoints (A. D. 1029) were first standard on acupoints compiled by the government. At modern times, standardization of Chinese acupuncture-moxibustion is being developed rapidly, twelve national standards have been issued including Nomenclature and Locations of Acupuncture Points and eleven standards on manipulation of acupuncture and moxibustion, and another fourteen standards on manipulation are still being developed. The organizations of standardization are established by Chinese Association of Acupuncture and Moxibustion (CAAM), and the standardization system and developing plan of acupuncture are compiled. The acupuncture standardization has put forward new questions before experimental research and clinical study of acupuncture moxibustion, and set a still higher requirement on training qualified personnel for acupuncture standardization research. PMID- 19186722 TI - [Protecting action of acupuncture serum on calcium over-loaded neurons of the hippocampus in rats of ischemia-reperfusion]. AB - OBJECTIVE: To investigate the effect of acupuncture serum on Ca2+ content in the cultured nervous cells of hippocampus after ischemia-reperfusion, so as to probe into humoral factors in acupuncture treatment. METHODS: The neurons of the hippocampus from the new born rats were cultured for 9-11 days. Fluorescein molecular probe Fluo-3 AM was used for staining of intracellular Ca2+. Fluorescent levels in the nervous cells cultured with the serum of the normal rats or the rats given electroacupuncture at "Baihui" (GV 20), "Zusanli" (ST 36), "Quchi" (LI 11) and "Sanyinjiao" (SP 6) for 2 weeks were determined by using a laser confocal microscope. RESULTS: After the normal serum was added, the intracellular Ca2+ fluorescent levels increased to 697 +/- 113 from 461 +/- 96, while after acupuncture serum was added, the Ca2+ fluorescent levels decreased to 584 +/- 103 from 673 +/- 108, indicating that after addition of acupuncture serum, the increased intracellular Ca2+ content could be decreased. CONCLUSION: There are some active substances in acupuncture serum which can obviously decrease intracellular Ca2+ content after ischemia-reperfusion, so as to provide a direct evidence for role of humoral factor in acupuncture treatment. PMID- 19186723 TI - [Effects of warm needle moxibustion on nerve root local inflammatory factors (NOS and CGRP) in the lumbar nerve root compress model rats]. AB - OBJECTIVE: To explore the mechanism of acupuncture for treatment of lumbar nerve root compression injury. METHODS: Fifty healthy SD rats were randomly divided into 5 groups, a normal group, a model group treated by saline, a medication group treated with Caerulein, an acupuncture group treated with acupuncture at L5, L6 Jiaji (EX-B 2) and a warm needle group treated with acupuncture and moxibustion at L5, L6 Jiaji (EX-B 2). The lumbar nerve root compress injury model was made by placing microsilica gel tablet. After they were treated for 14 days, the compressed nerve root was taken and the ultra-microstructure changes of the injured nerve root were observed by electron microscope and changes of nitric oxide synthase (NOS) and calcitonin gene-related peptide (CGRP) expressions were investigated by ELISA assay. RESULTS: The changes of ultra-microstructure of the nerve root were the most obvious in the model group and the changes in the medication group, the acupuncture group and the warming needle group reduced in order; the NOS activity and CGRP content in the nerve root tissue of the compressed area in the warm needle group were significantly reduced as compared with the model group (P < 0.05), but with no significant difference as compared with those in the normal group (P > 0.05). CONCLUSION: Warm needle treatment can effectively maintain cellular form, and ultra-microstructures of nerve root dorsal root ganglia, and effectively inhibit the release of inflammatory factors NOS and CGRP. PMID- 19186724 TI - [Preliminary study on disease menu of acupuncture and moxibustion abroad]. AB - Sum up clinical diseases and symptoms treated by modern acupuncture and moxibustion and disease menu of acupuncture and moxibustion abroad. By literature research methods, the diseases and symptoms reported in clinically acupuncture and moxibustion randomized controlled paper from 1978-2007 in databank of Pubmed network, USA, were analyzed and summarized, and the number of paper (frequency times) and conclusion of therapeutic effect (effective and ineffective) of each disease reported were counted up. Five hundred and eighty-seven papers of clinical acupuncture and moxibustion abroad, including 130 diseases and symptoms of 16 systems, were found. In these papers, 110 diseases and symptoms were unanimously considered effective and 16 diseases and symptoms were considered effective by most papers, and one disease was showed by equal number of paper of positive or negative effect, and one disease was considered ineffective by most of the paper, and 2 diseases and symptoms were considered ineffective by all of the papers. It is indicated that acupuncture and moxibustion have wide indications and have been widely applied at home and abroad, and disease menu of acupuncture and moxibustion is enlarging, and acupuncture and moxibustion have a vast application prospects. PMID- 19186725 TI - [Approach to clinical trial design for acupuncture and moxibustion treatment of stroke]. AB - To understand the differences of the clinical research of acupuncture at home and abroad in methodological design and implementation, so as to provide reference for exploring acupuncture clinical study design program suitable to China. Reading clinical research literature of acupuncture and moxibustion for treatment of stroke at home and abroad and analyze and probe the methodological design and implementation. The results indicate that clinical research of acupuncture and moxibustion abroad has advantages in the randomized, controlled and double blind, etc. We should adopt the advantages of Chinese medicine, enrich and perfect research design program which can be approved by modern medicine and properly reflect the therapeutic effect of acupuncture and moxibustion on stroke. PMID- 19186726 TI - [Differentiation and analysis on "keeping vitality"]. AB - Precise meaning of "keeping vitality" of acupuncture in Lingshujing * Nine Needling and Twelve origins is explained from the word source of "vitality", holding that "vitality" here is recognition of the ancients for delicate changes of pulse, and essence of "keeping vitality" is the course of understanding changes of qi and blood by pulse-taking. PMID- 19186727 TI - [Systematic assessment of acupoint stimulating therapy for deglutition disorders after apoplexy]. AB - OBJECTIVE: To assess the therapeutic effect of acupoint stimulating therapy on deglutition disorders after apoplexy, so as to provide basis for clinical treatment and further study. METHODS: According to Cochrane systematic assessment method of evidence-based medicine, Meta analysis on randomized controlled trials or half randomized controlled trials of acupoint stimulating therapy for deglutition disorders after apoplexy were made. RESULTS: In 37 Chinese literatures, 3 697 patients in total were enrolled, and the results indicated that the effective rate of acupoint stimulating therapy on deglutition disorders after apoplexy was better than that in the control group [RR was 1.38, 95% CI (1.28, 1.49), Z = 8.38, P < 0.01]; and the cured rate also was better than that of the control group [RR was 2.56, 95% CI (2.15, 3.04), Z = 10.70, P < 0.01]. CONCLUSION: The therapeutic effect of acupoint stimulating therapy on deglutition disorders after apoplexy is better than that of the control group, but more randomized, double blind, controlled trials with good designs are needed to confirm this result. PMID- 19186728 TI - [Progresses of studies on disease factors influencing the therapeutic effect of acupuncture and moxibustion on bronchial asthma]. AB - The authors preliminarily analyze and compare influence of different stages of attack and remission, different syndrome types, and age of the patient, duration of illness, state of illness on the therapeutic effects of acupuncture and moxibustion for treatment of bronchial asthma by analysis and summarization of 381 clinical papers about acupuncture and moxibustion treatment of bronchial asthma. It is found that most studies hold that acupuncture has better therapeutic effect on asthma at acute stage, and moxibustion has a better effect at the remission stage. Acupuncture and moxibustion have a better effect on cold asthma than hot asthma, and the shorter the course of disease and the younger, the better the effect, conversely, the effect is worse. Mild asthma is the main indication of acupuncture and moxibustion, but considerable more clinical studies show that above factors have no obvious influence on the therapeutic effect, even show contrary to this result. It is indicated that in the studies from now on, only strictly design and fully consider the effects of various factors, can scientifically reveal various disease factors influencing therapeutic effect of acupuncture and moxibustion on bronchial asthma, and expound the indications of different acupuncture and moxibustion methods for treatment of bronchial asthma. PMID- 19186729 TI - [Clinically randomized controlled trials of moxibustion with salt in bamboo circle for treatment of periarthritis of shoulder]. AB - OBJECTIVE: To assess the clinical therapeutic effect of moxibustion with salt in bamboo circle on periarthritis of shoulder. METHODS Eighty-six cases were randomly divided into a salt-moxibustion group and an electroacupuncture group, 43 cases in each group. The salt-moxibustion group was treated by moxibustion with salt in bamboo circle on the part of obvious pain and hot compress moxibustion on its periphery. The electroacupuncture group was treated with electroacupuncture at Jianyu (LI 15), Jianliao (TE 14), Jianzhen (SI 9), etc. After treatment, the analgesic effect and improvement degrees of active function of the shoulder joint were observed and 3 months later they were followed-up. RESULTS: The two therapies had analgesic effect and could improve active function of shoulder joint, but the salt-moxibustion group in the transient analgesic effect and the improvement degree of active function of the shoulder joint was better than the electroacupuncture group (P< 0.01, P < 0.05). Follow-up survey showed good clinical therapeutic effects in the two groups. The effective rates of pain and active function of shoulder joint were 97.7% and 93.0% in the salt moxibustion group and 93.0% and 88.4% in the electroacupuncture group, respectively, with no significant differences between the two groups. CONCLUSION: Moxibustion with salt in bamboo circle has an obvious therapeutic effect on periarthritis of shoulder, and it has transient analgesic effect and improves active function of shoulder joint, with a stable and long-term therapeutic effect. PMID- 19186730 TI - [Literature quality evaluation criteria for Clinical Practice Guideline of Evidence-based Acupuncture and Moxibustion]. AB - In Clinical Practice Guideline of Evidence-based Acupuncture and Moxibustion, following principles and methods of evidence-based medicine, in combination with characteristics of acupuncture and moxibustion science, primary literature quality assessment criteria and corresponding scale were stipulated and were repeatedly seeked advice from experts and proved, finally, forming the assessment criteria: (1) Evaluation criteria of literature quality for RCT; (2) Evaluation criteria of literature quality for non-randomly controlled trials; (3) Evaluation criteria of literature quality for cases-study trials. PMID- 19186731 TI - [Effect of acupuncture on blood oxygen saturation in patients of obstructive sleep apnea-hypopnea syndrome]. AB - OBJECTIVE: To investigate the effect of acupuncture on blood oxygen saturation in the patient of obstructive sleep apnea-hypopnea syndrome (OSAHS) in sleeping and to evaluate the therapeutic effect of acupuncture on this disease. METHODS: Thirty cases with OSAHS were treated with acupuncture at Shanglianquan (Extra), Fengfu (GV 16), Yamen (GV 15), Fengchi (GB 20), etc. 3-5 sessions each week. After treatment of 30 sessions, apneahypopnea index (AHI), mean blood oxygen saturation (MSaO2), the lowest blood oxygen saturation (LSaO2), oxygen desaturation > or = 4% index (ODI4), the mean blood oxygen saturation of oxygen desaturation when SaO2 < 90%, the longest time of oxygen saturation > or = 4% were observed before and after treatment. RESULTS: The effective rate of acupuncture was 23.3% for OSAHS. After acupuncture, AHI and ODI4 significantly reduced (P < 0.01); LSaO2 significantly increased (P < 0.01); MSaO2 and the mean blood oxygen saturation of oxygen desaturation when SaO2 < 90% significantly enhanced (P < 0.05); the longest time of oxygen saturation > or = 4% did not significantly change. CONCLUSION: The acupuncture treatment has intervenient effect on OSAHS and alleviates anoxia, so acupuncture is one of therapies improving anoxia in patients of OSAHS. PMID- 19186732 TI - E-prescribing: worth the fuss? PMID- 19186733 TI - CPT 2009. Out with the old, in with the new. PMID- 19186734 TI - Building the case for the patient-centered medical home. PMID- 19186735 TI - New year, new Medicare preventive coverage. PMID- 19186736 TI - E-prescribing: why the fuss? PMID- 19186737 TI - Charting then and now. PMID- 19186738 TI - Environmental payoff. PMID- 19186740 TI - Big little problem. PMID- 19186739 TI - Virus in the brain. PMID- 19186741 TI - Space sticker shock. PMID- 19186742 TI - Neighborhood darkness. PMID- 19186743 TI - Decoding the mammoth. PMID- 19186744 TI - A theory for everyman. PMID- 19186745 TI - Blackouts and cascading failures. PMID- 19186746 TI - Telephone to the dead. PMID- 19186747 TI - Darwin's living legacy. PMID- 19186748 TI - Testing natural selection. PMID- 19186749 TI - From atoms to traits. PMID- 19186750 TI - The human pedigree. PMID- 19186751 TI - This old body. PMID- 19186752 TI - What will become of Homo sapiens? PMID- 19186753 TI - Four fallacies of pop evolutionary psychology. PMID- 19186754 TI - Evolution in the everyday world. PMID- 19186755 TI - The science of spore. PMID- 19186756 TI - The latest face of creationism. PMID- 19186757 TI - A theory of a deadly fusion. PMID- 19186758 TI - New designs going up. PMID- 19186759 TI - How does solar power work? PMID- 19186760 TI - Cancer in circumpolar populations. PMID- 19186761 TI - Cancer among the circumpolar Inuit, 1989-2003. I. Background and methods. AB - OBJECTIVES: This is first of 2 papers on the Circumpolar Inuit Cancer Review, an international collaborative effort involving researchers and health officials from Alaska, Canada and Greenland. It covers the period 1989-2003, updating the last review (1969-1988) and together provides an overview of the trends and patterns of cancer among the Inuit in 3 countries and over a 35-year period. METHODS: Inuit cancer cases by age-sex group and anatomic site were obtained from the regional cancer registries. The sources of the age-sex distribution of various Inuit populations include the population registry (Greenland), and annual estimates and periodic censuses (Alaska and Canada). Incidence rates were age standardization by the direct method to the standard world population of the International Agency for Research on Cancer. CONCLUSIONS: This project demonstrates the feasibility of international partnerships in cancer surveillance, and when these partnerships are extended to other diseases and health conditions, they can contribute to the development of a Circumpolar Health Observatory. PMID- 19186762 TI - Cancer among the circumpolar Inuit, 1989-2003. II. Patterns and trends. AB - OBJECTIVES: This is the second of 2 papers reporting on the result of the Circumpolar Inuit Cancer Review, an international collaborative effort involving researchers and health officials from Alaska, Canada and Greenland. METHODS: Inuit cancer cases by age-sex group and anatomic site were obtained from the regional cancer registries. RESULTS: Cancer in general is increasing among Inuit, in all regions, and among both men and women. Inuit continue to be at extreme high risk, relative to non-Inuit and to comparisons of global populations, for the historically recognized so-called traditional cancers (such as cancer of the nasopharynx and salivary glands). Among the so-called modern cancers prevalent in developed societies, lung cancer is rapidly increasing in incidence (especially in Canada), such that the rate in both Inuit men and women is the highest in the world; other cancers, such as colorectal cancer, are also on the rise (especially in Alaska), while breast and prostate cancer remain low relative to the non-Inuit population. The decline in cervical cancer is a positive development; in the 3 regions, the rate in Greenland is the highest. CONCLUSIONS: Data such as these can form the basis of interventions directed towards known risk factors such as smoking, diet, obesity, viral and bacterial infections, and low screening prevalence. Cancer surveillance is a basic task of the public health system; in the Arctic, it is particularly important as Inuit continue to undergo further changes in their life-styles and social environments. PMID- 19186763 TI - Cancer among the Sami--a review on the Norwegian, Swedish and Finnish Sami populations. AB - OBJECTIVES: The Sami are the Indigenous people of the northernmost parts of Sweden, Finland and Norway, and of the Kola Peninsula of Russia. The present review summarizes the main results from studies on cancer morbidity and mortality among the Sami and discusses these results in relation to exposure of known risk factors. STUDY DESIGN: Literature review. METHODS: A systematic search over the time period 1966-2008 for relevant articles was conducted on MEDLINE. Updates and recalculations of some of the results from the original data were also done. RESULTS: Nine articles whose main focus is on cancer incidence or mortality among the Sami were identified. In all studies, the overall incidence of cancer or cancer mortality was lower among the Sami in comparison with the national populations. The differences were less striking in relation to regional reference populations, but the rates were still significantly lower for all populations of Sami, except for Swedish Sami women. Beyond the general trend of a lower cancer incidence among the Sami, there were some notable differences between the various Sami subpopulations. CONCLUSIONS: The risk of developing and dying from cancer is low among the Sami. A life-style that includes cancer-protective factors, such as certain dietary components and physical activity, is the most likely explanation for the lower incidence of cancer among the Sami. PMID- 19186764 TI - Cancer incidence in the Nenetskij Avtonomnyj Okrug, Arctic Russia. AB - OBJECTIVES: To assess the overall and main site-specific cancer incidence in Nenetskij Avtonomnyj Okrug (NAO). STUDY DESIGN: A register-based analysis of incidence in the period 1993-2006. METHODS: NAO, a part of Arkhangelskaja Oblast in north-west Russia, has a population of 42,000 inhabitants. The central oncological hospital of the oblast registers all new cases of cancer. All new cases recorded in the study period among official residents of NAO were included in the study, except for secondary malignant neoplasm, cases revealed by autopsy and cancers diagnosed within 6 months of a previous cancer diagnosis. The census and annual sex and age-group-specific population figures for NAO were obtained from the regional statistics office. Crude and age-adjusted incidence rates (to the world standard population) were estimated. RESULTS: The average crude cancer incidence per year was 204/100,000 among men and 194/100,000 among women. Adjusted for age, the incidence was 322/100,000 and 182/100,000, respectively. The most frequent primary site of cancer was trachea, bronchus and lung, which constituted 17.3% of all cases (of which 87% were among men), followed by stomach cancer (12.5%). Breast cancer constituted 17.5% of all cases among women. CONCLUSIONS: The results are consistent with reports of a low cancer risk among women compared with men in Russia and compared with women in Western countries and with results that point out that public health measures are needed to curb the lung cancer epidemic among men in Russia. The high risks of pancreas, kidney and oesophagus cancers among men should be investigated further. PMID- 19186765 TI - Cancer incidence and mortality among Aboriginal people living on reserves and northern villages in Quebec, 1988-2004. AB - OBJECTIVES: Little information is available on the incidence and mortality of cancer among the Aboriginal population in the Province of Quebec, Canada. Cancer was likely rare in this population historically, but recent life-style changes suggest that this may no longer be the case. The purpose of this study was to estimate incidence and mortality rates among Aboriginal people living on reserves and in northern villages in Quebec during the period 1988-2004, and to compare these estimates with those of the general population. STUDY DESIGN: Incidence and mortality data were extracted respectively from the provincial tumour registry and death file. METHODS: Aboriginal people were identified based on geographic residence codes. Population data were taken from the Canadian census of 1991, 1996 and 2001. Incidence and mortality rates were calculated and age-standardized according to the World Standard Population. RESULTS: The Aboriginal incidence and mortality rates for cancer, all sites combined, was 321.8 per 100,000 (95% confidence interval 304.5-339.2) and 160.3 per 100,000 (95% CI 147.8-172.8), respectively. These rates are not significantly different from those of the general population of Quebec. However, there are differences according to cancer site and sex. Aboriginal men had a higher risk for liver, lung and kidney cancers and a lower risk for prostate, bladder, leukemia and non-Hodgkin's lymphoma cancers, whereas Aboriginal women had a higher risk for colorectal, lung, cervix and kidney cancers, and a lower risk for breast, uterus, bladder, brain, leukemia, stomach and pancreas cancers. CONCLUSIONS: Aboriginal people in Quebec now experience an overall cancer risk comparable to the general population. The observed differences in specific sites suggest exposure to unique environmental risk factors. Basic surveillance specific to this population is necessary for the planning and evaluation of cancer preventive and curative services. PMID- 19186766 TI - Hearing outcomes after mobile ear surgery for chronic otitis media in Greenland. AB - OBJECTIVES: Chronic otitis media (COM) with hearing loss (HL) occurs frequently in many populations with limited access to specialized treatment. This article reports on the hearing outcomes following mobile ear surgery in Greenland. STUDY DESIGN: The study was longitudinal and prospective with pre- and post-operative followup data compiled at 1- and 2-year intervals. METHODS: Selection criterion included individuals with COM with or without suppuration (CSOM) and with associated HL. Hearing tests were obtained by using a simple air-conduction (AC) audiometry. The median age of the 274 participants was 27 years (8-60 years), and 45% were males. Sixty-one percent had COM and 34% had CSOM. Myringoplasty or tympanoplasty type I was performed in 88% of the cases. RESULTS: The median pre operative AC pure-tone average was 38 dB. The follow-up rate was 75% and 61% at 1 and 2-years. Hearing gain was found in 78% at both follow-ups and was > or = 10 dB in 62% and 56%, respectively. The median hearing gain was 15 dB and 12 dB, respectively. Independent predictors of hearing gain were pre-operative tympanic membrane atrophy and closure of perforations. CONCLUSIONS: Long-term hearing gain can be achieved with mobile ear surgery and this, in addition to tympanic membrane closure, is important for hearing rehabilitation in populations with poor access to health care. PMID- 19186767 TI - Experiences of having a prematurely born infant from the perspective of mothers in northern Sweden. AB - OBJECTIVES: The aim of this study was to describe the mothers' experience of having a prematurely born infant, with the focus on the birth itself and during the time immediately following the birth. STUDY DESIGN: A qualitative design was chosen to achieve the aim of this study. METHODS: Six mothers, whose infants were born prematurely and thereby needed care on a neonatal intensive care unit (NICU), participated in the study. Narrative interviews were conducted and the mothers were encouraged to freely narrate their experiences of having an infant born preterm. A qualitative content analysis was used to analyse the interviews. RESULTS: The results show that mothers were not prepared for having prematurely born infants and that initially they had difficulties feeling like a mother. Having an infant born preterm was dominated by feelings of anxiety. Feeling closeness to the child was important, and separation from the child was a very stressful experience. Family life was affected, but mothers felt they were able to handle the situation if they received support from their partner and hospital staff, and if they were equipped with knowledge regarding the birth and care of a premature infant. Furthermore, it was important for the mothers to be involved in their infants' care. CONCLUSIONS: Support and knowledge made it possible for mothers to handle having a premature infant. Mothers' situations could be facilitated if nurses had increased knowledge and understanding about how mothers experience this situation. PMID- 19186768 TI - Expectations of post-partum care among pregnant women living in the north of Sweden. AB - OBJECTIVES: To describe expectations of post-partum care among pregnant women living in the north of Sweden and whether personality determines preference for care systems. The time for post-partum care on maternity wards has been reduced in Western countries. This, along with the reduction in special medical treatments offered and the closure of small hospitals has affected pregnant women and their families. STUDY DESIGN: Data was extracted from a questionnaire and a personality instrument (SSP) that were completed during November 2002. METHODS: In the northernmost county of Sweden, 140 pregnant women completed the questionnaire; of these, 120 completed the SSP instrument. RESULTS: Of the women who participated, 61.3% wanted to be discharged 72 hours after childbirth, irrespective of the distance between the hospital and home. To have access to maternity ward staff and the decision to be discharged were described as being the most important issues in maternity ward care. The infant's father was expected to be the most important person in the post-partum period. CONCLUSIONS: Women ranked the opportunity to decide for themselves when to be discharged from the maternity ward as important, which can be interpreted as a strong signal that the women want to be in control of the care they receive. Midwives have to focus more on the woman and her family's individual needs, and to include the father as a person who also needs support and to provide resources for him. PMID- 19186769 TI - Access redux. PMID- 19186770 TI - Apert syndrome: report of a case with emphasis on craniofacial and genetic features. AB - Apert syndrome is 1 of the 5 craniosynostosis syndromes that shore clinical features and are caused by allelic mutations in the fibroblast growth factor receptor 2 (FGFR2) gene. The purpose of this paper was to report a case of Apert syndrome, with particular emphasis on craniofacial and genetic features, in a 5 year-old female patient. The patient presented with several craniofacial deformities, including severe brachycephaly, midface hypoplasio, flat forehead, proptosis, hypertelorism, and short nose with a bulbous tip. Syndactylies of the hands and feet were also observed. Intraoral findings included arched palate with pseudocleft in the midline, upper lip with symmetric depression resembling pseudoclefts, severe malocclusion, and several decoyed teeth. DNA sequence and restriction enzyme analysis showed a G to C transversion, resulting in a serine to tryptophan amino acid substitution at position 252 (S252W). Identification of the clinical features associated with mutation analysis is important to correctly diagnose Apert syndrome and distinguish it from other clinically similar craniosynostosis syndromes. PMID- 19186771 TI - Failure rates and survival analysis of maxillary expansion appliances: a 24-year retrospective analysis. AB - PURPOSE: The purpose of this study was to describe the failure rates of maxillary expansion appliances (MEAs) and assess risk variables associated with failures and treatment time. METHODS: Retrospective chart reviews were performed on 436 primary or mixed dentition patients presenting with crossbite to a private practice between 1981-2005. Survival analysis was used to analyze and compare the types of MEAs with respect to the treatment time. The effect of demographic and clinical characteristics on appliance failure and treatment time was assessed using linear and logistic regression models. RESULTS: The average age at insertion of a MEA was 8 years, 4 months (+/-1.72 SD). Nineteen percent (n=84) of the MEAs failed, with a median treatment time for all appliances of 216 days (interquartile range=126 days). Cement loss (69%) was the most common type of failure. The likelihood of an appliance failing increased in children with a malocclusion other than Class I (adjusted odds ratio=1.91; 95% CI=1.16-3.14) and was nearly 4 times greater when a quad helix was used compare to the Haas appliance (adjusted odds ratio=3.60; 95% CI=1.92-6.75). The treatment time was significantly affected by the type of crossbite present and the occurrence of an appliance failure (P=.001). CONCLUSIONS: The use of a quad helix appliance and the presence of malocclusion other than Class I was significantly predictive of appliance failure. Treatment time was increased when MEAs failed and bilateral crossbite was present. PMID- 19186772 TI - Tobacco control in pediatric dental practices: a survey of practitioners. AB - PURPOSE: The study purpose was to determine tobacco-related knowledge, attitudes, and practice behaviors of American Academy of Pediatric Dentistry (AAPD) members. METHODS: A 26-item survey was distributed to a national, random sample of 1,700 AAPD members. Frequencies, odds ratios and 95% confidence intervals assessed factors related to tobacco control behaviors. RESULTS: Of 1,700 questionnaires, 1,292 (82%) were returned and usable. Over 75% of respondents agreed that it is a pediatric dentist's responsibility to help patients who wish to stop using tobacco; only 142 (11%) had prior tobacco prevention/cessation training. Of those untrained, 905 (70%) were willing to be trained. Not knowing where to send patients for counseling and feeling ineffective with helping patients to stop their tobacco use were significant barriers reported by nearly half the respondents. Two hundred forty-five (19%) reported always/often asking their adolescent patients about tobacco use; 491 (38%) reported always/often advising known tobacco users to quit; and 284 (22%) reported always/often assisting with stopping tobacco use. Feeling well prepared to ask about tobacco was significantly associated with assisting tobacco users (odds ratio=8.9; 95% confidence interval=6.6-12). CONCLUSION: Continuing education programs are needed to enhance the knowledge and skills of pediatric dentists to promote tobacco control behaviors. PMID- 19186773 TI - Caregiver's perception of child's oral health status among low-income African Americans. AB - PURPOSE: This study aimed to: (1) compare caregivers' perceptions of their children's oral health status with clinical findings; and (2) investigate the influence of caregivers' attitudes, beliefs, and knowledge concerning dental caries development and oral health on caregivers' perception levels of their children's oral health status. METHODS: A representative sample of low-income African American families (0- to 5-year-olds and their caregivers) in Detroit, Mich was interviewed. Caregivers were asked to assess their own and their children's oral health status. All children and their caregivers received dental examinations. This study included data from 885 caregivers with children aged 1 year or older. RESULTS: Approximately 79% of caregivers rated their children's oral health as good to excellent, and 21% rated it as fair to poor. Caregivers' perception of their children's oral health was significantly associated with their children's caries experience, as measured by the number of decayed, missing, and filled tooth surfaces. It was also associated with limitations of oral functions, such as chewing difficulty. Poorer perceptions of caregivers' oral health and fatalistic attitudes toward children's oral health were significantly associated with poorer perception of their children's oral health. CONCLUSION: Caregivers' perception of their children's oral health status is a significant indicator of the children's clinical caries experience. PMID- 19186774 TI - Nutritional counseling practices: how do North Carolina pediatric dentists weigh in? AB - It has been demonstrated that nutrition/healthy lifestyle counseling can help alleviate the childhood obesity crisis. Because pediatric dentists see children with regular frequency, they are uniquely positioned to help provide such counseling. PURPOSE: To document current nutrition/healthy lifestyle counseling practices of pediatric dentists in NC and examine factors associated with those practices. METHODS: A 65-item survey instrument was developed by an expert committee, pilot-tested and revised. The instrument contained questions in 6 domains: Academic Preparation, Knowledge, Confidence, Opinions, Practice Patterns and Barriers. It was sent to all clinically active pediatric dentists in NC. RESULTS: The response rate was 69% (70/102). Less than 25% provided nutrition counseling services. Academic preparation in residency training was statistically significant (P<.05) in determining those who practiced nutritional counseling. Logistic regression revealed that respondents with higher overall knowledge (OR=18.2), those comfortable discussing weight-related issues (OR=47.7), those in practice >10 years (OR=20.6), and females (OR=173) were more likely to provide nutrition/healthy lifestyle counseling. CONCLUSIONS: All respondents believed that childhood obesity is a major health concern and expressed a willingness to assist other health professionals in addressing this problem. These findings offer a rich source of data that can be used for educational initiatives and strategic planning. PMID- 19186775 TI - Evaluation of the Root ZX apex locator in primary teeth. AB - PURPOSE: The purpose of this study was to evaluate the accuracy of the Root ZX electronic apex locator in determining canal length in primary incisors. METHODS: Root ZX was utilized to measure the canal lengths of 150 primary incisors in vivo. After the teeth were extracted, a standard ruler was used to measure the canal lengths to the nearest 0.5 mm. A paired t test was used to compare the lengths obtained in vivo with Root ZX and the lengths measured in vitro. RESULTS: In 98 (65%) of the canals, the length measured by Root ZX was determined to be equal to the length measured in vitro. Root ZX measurements were found to be less than in vitro measurements by 0.5 mm in 24 canals (16%), and were greater than in vitro measurements by 0.5 mm in 21 canals (14%). Root ZX measurements exceeded in vitro measurements by 1.0 to 1.5 mm in 5 canals (3%) and were less than in vitro measurements by 1.0 mm in 2 canals (1%). Paired t test analysis indicated no significant difference between the 2 measurement methods (P=.67). CONCLUSIONS: Root ZX can accurately assess the length of canals in primary incisors. PMID- 19186776 TI - Evaluation of an early childhood caries prevention program at an urban pediatric clinic. AB - PURPOSE: The purpose of this study was to evaluate a risk-based dental caries prevention program at an urban pediatric primary care clinic serving low-income residents. METHODS: A demonstration caries prevention trial lasting 26 months was conducted with 219 6- to 27-month-old children. A "prevention" group of 6- to 15 month-old children (initial visit) were compared at the end of the trial with a "comparison" group at their initial visit. The latter children were patients of the same clinic who were 12 months older (18-27 months). Preventive services included: caries-risk assessments; mutans streptococci (MS) monitoring; fluoride varnish applications; dental health counseling; referral for dental treatment, if indicated; and periodic recalls. RESULTS: Prevention group children at the last recall experienced fewer mean carious dental surfaces (0.1 vs 1.29, P=.01) and over 8-fold less MS (P=.01) than comparison group subjects at their initial visit. The number of precavitated lesions was not significantly different CONCLUSIONS: Administration of prevention measures was shown to reduce dental caries experience in low socioeconomic status infants and toddlers, and oral MS levels were a reliable indicator of future caries status. Outcomes support establishment of caries prevention programs at urban primary care pediatric clinics. PMID- 19186777 TI - Association between early childhood caries and behavior as measured by the Child Behavior Checklist. AB - PURPOSE: This study's purpose was to use a standardized behavioral assessment instrument to compare the observed behavior of caries-free (CF) with caries active (CA) children requiring restoration under general anesthesia (GA). METHODS: Healthy 30- to 60-month-olds, either CF or in need of dental restoration under GA, were selected in 4 equal-size cohorts of 30 subjects from both private practice settings and a clinic. Trained assistants administered the child behavior checklist, a standardized, age-appropriate instrument to assess child behavior problems to parents at the time of appointment. RESULTS: The 60 CF and 60 CA subjects did not differ significantly (P<.05) between clinic and private practice for age, gender, and race and were collapsed into groups of 60 CF and 60 CA children (mean ages=41.2 and 45.2 months, respectively). Compared to CF children, CA children had significantly higher (more problems) scores for categories of: anxious/depressed; sleep problems; aggressive behavior; externalizing and total problems; and attention deficit/hyperactivity problems. CA children scores for withdrawn, sleep problems, and attention deficit/hyperactivity problems were also higher than normal, but neither group had scores outside normative ranges. CONCLUSIONS: No significant age, gender, behavior, or race differences were noted for site of care, but caries-active children had significantly more behavior problems than caries-free children. PMID- 19186778 TI - Neuropathic orofacial pain in children and adolescents. AB - The field of orofacial pain is broad and includes pain conditions affecting the head, face, neck, and intraoral structures. Much has been written about orofacial pain, specifically chronic pain conditions affecting adults. Little is known, however, about the presentation of orofacial pain in pediatric patients. The objective of this paper was to increase awareness and provide information regarding neuropathic orofacial pain in children and adolescents, as pediatric dentists may be asked to participate in the treatment of these patients. Moreover, an accurate history and examination can lead to on early diagnosis of the pain condition, thus avoiding unnecessary or inappropriate dental procedures. PMID- 19186779 TI - Articaine use in children among dental practitioners. AB - PURPOSE: The purpose of this study was to examine local anesthetic selection and dentists' use of articaine in children. METHODS: Using a cross-sectional survey design, a questionnaire regarding the use of local anesthetics in children was mailed to a random sample of dentists and all pediatric dentists from North Carolina and Virginia. The 16-item questionnaire included questions regarding the preferred local anesthetic used in children. The association between dental practitioner type and anesthetic use was tested using the chi-square or Fisher's exact test. RESULTS: There was a 30% response rate. Lidocaine with epinephrine was the local anesthetic preferred overall by all practitioners. Approximately 50% of dentists surveyed reported using articaine in children. There were no significant differences in the preference of articaine between provider types with the exception of older, 7- to 10-year-old patients, where general dentists preferred articaine significantly more than pediatric dentists (28% vs 16%). CONCLUSIONS: While lidocaine with epinephrine was still the preferred local anesthetic for use in children, the use of articaine in children was prevalent among both general and pediatric dentists. The use of articaine became more prevalent as the patient's age increased. PMID- 19186780 TI - Physiologic signs during dental treatment in overweight vs normal weight children. AB - PURPOSE: This study compared physiologic effects of position change on overweight (OW) and normal-weight (NW) children during a preventive appointment. METHODS: A cross-sectional, case-controlled design assigned a convenience sample to either overweight (>85th percentile) or normal-weight groups (25th to 85th percentile) by body mass index (BMI). A trained examiner measured blood pressure (BP), heart rate (HR), oxygen saturation (SpO2), and 5 pulmonary functions in preoperative, upright seated, supine, and postoperative upright seated positions. Anxiety was measured using the Corah Dental Anxiety Scale. RESULTS: NW and OW groups' anxiety scores did not differ (P=.69). Mean BMIs were 20.1 and 39.7 for NW and OW subjects, respectively. Overall, BP between groups was not different (P=.051), although systolic BP for OW subjects tended to be higher. Systolic and diastolic BP dropped for both groups with supine positioning (P< or =.001). During this change, HR did not change for NW, but dropped for OW subjects (P=.001). SpO2 was significantly different between groups following position change, but not within each group. Forced vital capacity and forced expiratory volume were higher in the OW group (P< or =.01), but 2 peak expiratory measures (PEF and PEF25-75 were not (P=.05). CONCLUSION: Heart rate and oxygen saturation respond differently to positional changes in overweight patients. PMID- 19186781 TI - A nasopalatine duct cyst in a 7-year-old child. AB - The nasopalatine duct cyst (NPDC) is a developmental cyst of the anterior palate's midline, usually presenting as an asymptomatic swelling located just behind the maxillary central incisors. It is the most common non-odontogenic cyst of the jaws but is seen rarely in children. The purpose of this paper was to report an unusual case of nasopalatine duct cyst in a 7-year-old boy who presented with a slow-growing, slight swelling of the anterior palate together with malpositioned permanent maxillary central incisors. Although rare in children, NPCD should be included in the differential diagnosis of anterior palate swelling, particularly if associated with malpositioned maxillary central incisors. PMID- 19186782 TI - Influencing health policy for the imminent health-care crisis: a task for informed citizens, proactive nurses, and committed researchers. PMID- 19186783 TI - Ethics in a time of contagion: a relational perspective. AB - In times of contagion, the key role of nurses brings fears, dangers, and unique demands. The ethics of such challenges need to be explored and understood. Using Callahan's framework for thinking ethically and Taylor's "worries" of modern life, the author elucidates some of the challenges and then argues that the current approach to pandemic ethics, with its reliance on moral reasoning, is insufficient to guide nurses' ethical actions. Relational ethics, which explicitly situates ethics within relationships and our commitment to one another, and which recognizes that context matters in ethical decision-making, is offered as a viable alternative for nurses in considering how to respond. PMID- 19186784 TI - Discovering the literature on Aboriginal diabetes in Canada: a focus on holistic methodologies. AB - Promoting culturally competent care for diabetes is critical to addressing the health inequities of indigenous peoples. Based on a review of studies published between 1995 and 2007, the author presents a descriptive analysis of Aboriginal diabetes knowledge guiding the involvement of advanced practice nurses in programs of care. While the literature review is not systematic, it is sufficiently comprehensive to provide a theoretical backdrop to Aboriginal diabetes programs of practice and research. In terms of a particular area of Aboriginal diabetes literature, it also provides a snapshot of community-based diabetes research focused on holistic methodologies historically used in the Canadian context. The Aboriginal diabetes literature is found to be a source of rich information. Analysis of holistic methodologies, however, reveals underdevelopment of research and limited use of key holistic guidelines for Aboriginal policy research. PMID- 19186785 TI - Actuarial assessment of violence risk in hospital-based partner assault clinics. AB - Hospital-based partner assault clinics are a relatively recent addition to the community response to partner violence. In this study, 66% of 111 women attending hospital clinics for partner assault were physically injured and 43% reported death threats. Few concurrently used other services (shelters or police) and most relied on female friends and relatives for help. Many participants who currently lived with the perpetrator were contemplating leaving but only a third had made plans to do so. Participants faced an unusually high risk of future assault, according to both victim interview using the ODARA actuarial risk assessment and their own perceptions. Findings imply an important role for partner assault clinics and the feasibility of the victim service sector's using the same actuarial risk assessments as the criminal justice system. PMID- 19186786 TI - Examination of negative affective responses to waiting for surgery. AB - Long surgical waiting times are associated with many negative consequences. Although depression and anxiety have been studied among patients waiting for surgery, fundamental anxieties, such as anxiety sensitivity and health anxiety, have not been examined. Furthermore, research into patients' perspectives on coping while waiting is limited. The purpose of this study was to examine the extent of anxiety sensitivity, health anxiety, depression, and anxiety and their influence on coping, concern about waiting, and anxiety about surgery in individuals waiting for surgery. Individuals waiting for surgery completed measures assessing these constructs. Results revealed a high degree of health anxiety and anxiety sensitivity. Regression analysis showed that health anxiety and depression are unique predictors of coping and together help to predict concern about waiting. Anxiety sensitivity and anxiety did not add to the prediction of these variables. Overall, the results suggest that further attention should be given to interventions that address psychological distress during the wait for surgery. PMID- 19186787 TI - Community resiliency as a measure of collective health status: perspectives from rural communities. AB - Community resiliency is a theoretical framework useful for describing the process used by communities to address adversity. A mixed-method 2-year case study was conducted to gather information about community resiliency in 2 rural communities. This article focuses on the themes generated from qualitative interviews with 55 members of these communities. The participants viewed community as a place of interdependence and interaction. The majority saw community resiliency as the ability to address challenges. Characteristics included physical and social infrastructure, population characteristics, conceptual characteristics, and problem-solving processes. Barriers included negative individual attitudes and lack of infrastructure in rural communities. Nurses could play a key role in enhancing the resiliency of rural communities by developing and implementing programs based on the Community Resiliency Model, which was supported in this study. PMID- 19186788 TI - The psychosocial work environment and evidence utilization by health professionals. AB - The purpose of this study was to investigate the relationships between dimensions of the psychosocial work environment and health professionals' use of evidence in their practice. A correlational descriptive design was developed. Health professionals working in mental health units at 2 hospitals were asked to complete a questionnaire about their perceptions of the psychosocial work environment and their use of evidence. Correlations and regression analyses were performed. Use of evidence was found to be correlated with social support and decision latitude. Results of multiple regression analyses found perceived social support (beta = .27, p < .01) and perceived decision latitude (beta = .25,p < .01) to be significant predictors of the use of evidence. The authors conclude that good social support and decision latitude among interprofessional groups may promote use of evidence by health professions in their practice. PMID- 19186789 TI - Interpretive hermeneutics and modifying the modern idea of method. AB - A challenge currently facing human sciences researchers is recognizing the importance of creating a shared horizon of meaning in their work. To move towards this goal, researchers require an awareness of how their chosen conceptual framework creates a stricture through which they know and understand the individual. The methodological approach of philosophical interpretive hermeneutics emphasizes that what is learned from experience extends beyond the strictures of formalized method, thus offering a middle way of thinking in the research encounter. This article explores how interpretive hermeneutics can broaden the notion of research from one of simply knowing to one of understanding. It does so by engaging with Gadamer's philosophical hermeneutics, where researchers strive to embrace a constellation of philosophical outlooks rather than a specific philosophical method. A key insight in research involving philosophical interpretive hermeneutics is its emphasis on human experience. PMID- 19186790 TI - Quirk-e: Queer seniors creating community. PMID- 19186791 TI - Taking stock: the first decade of the Nursing Research Fund. PMID- 19186792 TI - [Nitrate pollution in groundwater for drinking and its affecting factors in Hailun, northeast China]. AB - Nitrate pollution in groundwater has become a worldwide problem. It may affect the water quality for daily use and thus the health of people. The temporal and spatial characteristics of nitrate pollution in the groundwater were addressed by sample analysis of the drinkable water from 157 wells in Hailun, Heilongjiang, northeastern China. It was found that the mean value of nitrate concentration in all wells was 14.01 mg x L(-1). Of all the samples, the nitrate concentrations of 26.11% wells exceeded the standard of drinkable water (10.00 mg x L(-1)). A significant difference was found on the spatial distribution of nitrate pollution in the study area. The pollution degree in term of nitrate pollution was in the order: the central rolling hills and flooding plain > the northeastern mountain area > the southwest rolling hills and plain. Based on the results, the factors causing the pollution we analyzed from the well properties and pollution sources. As for well properties, the type of the pipe material plays a critical role in the groundwater nitrate pollution. It was found that the wells with seamless pipe have less pollution than those with multiple-sections pipe. The concentrations of seamless pipe wells and multiple ones were respectively 5.08 mg x L(-1) and 32.57 mg x L(-1), 12.26% and 82.35% of these two kinds wells exceeded 10.00 mg x L(-1), the state drinking water standard. In the whole Hailun, there is no statistically relationship between nitrate-N levels of wells and the well depth. However, a statistically lower nitrate-N was observed in the deep wells than that in the shallower ones. The mean values of nitrate concentration of the seamless-pipe deep wells, seamless-pipe shallow wells, multiple-section-pipe deep wells and multiple-section-pipe shallow wells were 1.84, 12.02, 25.14 and 45.61 mg x L(-1). Analysis of pollution source shows that the heavily polluted regions are usually associated with large use of nitrogen fertilizer and household livestock or poultry. This indicates a positive correlation between the nitrate-N pollution of groundwater and the nitrogen fertilizer, household livestock, poultry. PMID- 19186793 TI - [Canonical correspondence analysis between phytoplankton community and environmental factors in winter and summer in shallow lakes of plain river network areas, Suzhou]. AB - Field investigations on the phytoplankton community were carried out in December 2005 and June 2006 in shallow lakes of plainriver network areas, Suzhou City. Results show that there are 73 species, 62 genus, 8 phylum phytoplankton, which is mainly composed of the Chlorophyta, Bacillariophyta and Cyanophyta. In winter, the average phytoplankton abundance is 254.88 x 10(4) cells x L(-1), and the Bacillariophyta abundance accounts for 62.3% of the total. While in summer, the average phytoplankton abundance is 2704.28 x 10(4) cells x L(-1), and the Cyanophyta abundance accounts for 93.5%. The relationship between 62 genus of phytoplankton and 11 environmental factors from 57 sampling sites in research area was studied by Canonical Correspondence Analysis. It suggests that water temperature, permanganate indexes, nitrate nitrogen and total nitrogen are the main environmental factors correlated with the distribution of phytoplankton community. And in winter, pH, ammonia nitrogen and total phosphorus are also the main environmental factors. Furthermore, the Bacillariophyta has higher adaptability to the changeable environment. The Chlorophyta can tolerate higher concentration of permanganate indexes, phosphorus nutrients, nitrogen nutrients and total organic carbon, whereas the response of Cyanophyta to environmental factors needs further research. PMID- 19186794 TI - [Major ionic features and their sources in the Nam Co Basin over the Tibetan Plateau]. AB - A preliminary study of major ions in water of three rivers (Niyaqu, Qugaqie, Angqu) in the Nam Co Basin has been carried out in June-October 2006. An obvious difference is found among these rivers, whose ionic characteristics are mainly influenced by the length of rivers and their supply sources. Among the three rivers, the contents of total dissolve solids (TDS) is 79.48 mg/L, 23.44 mg/L, and 111 mg/L in the Niyaqu, Qugaqie, and Angqu, respectively. The hydro-chemical type is CCa for the Qugaqie, between CNa and CCa for the Angqu, and between the Angqu and Qugaqie for the Qugaqie. Changes in ionic concentrations are also related with discharges of the rivers. According to principal component analysis and correlation analysis, the main chemical contents derive from Ca2+ and HCO3(-) in carbonate for the Niyaqu, glacier meltwater and carbonate for the Qugaqie, and Na+, Mg2+ and Cl(-) in evaporates for the Angqu. PMID- 19186795 TI - [Temporal and spatial variation rule of methylmercury in water in the Second Songhua River]. AB - The concentrations of methylmercury (MeHg) in water were investigated in both level period and flood period in the Second Songhua River. The results show that the MeHg concentrations in water of Hongshi, Shihaoxian and Ganshuigang sites are relatively higher than other sampling sites. The MeHg concentrations in the level period are higher than the flood season in most sampling sites beside Laozhou and Sifangtai sites. By comparing the MeHg concentrations in different years, the results reveal that the MeHg concentrations in 1981 decreased with the distances increasing from the sites to the pollution source. The MeHg concentrations in most sites greatly fell in 1983, when the pollution source had been ceased for one year. The concentrations approached the natural background level in most sites, however, they were over 0.35 ng x L(-1) from Zhaoyuan site to Sifangtai site. The MeHg concentrations in corresponding sites in 2006 were higher than that in 1983, but were lower than that in 1981. Moreover, the pollution level in water of the Second Songhua River was also discussed. PMID- 19186796 TI - [Remote sensing inversion mode of suspended particles concentration in Hangzhou Bay based on in situ measurement spectrum]. AB - Suspended particles are one of major parameters of coastal water color remote sensing in China. The reflectances spectral of water were measured using an ASD field spectroscope, synchronously the suspended particles concentrations of surface water were acquired in Hangzhou Bay. Two remote sensing inversion models for suspended particles concentrations (SPC) were developed based on t hesimulated reflectance of MODIS & MERIS channels using artificial neural network (ANN) algorithm. Measurement results show that the total SPC of Hangzhou bay is comparatively high where the suspended sediments concentrations (SSC) are far more than chlorophyll concentrations, averagely 705 mg/L and 1.164 mg/m3, reseparately. The SPC in two measurement stations appears spatio-temporal variation, especially the short period change due to tidal cycle. There are two reflectance peaks in the measured spectral curves, one is between 650 nm to 750 nm, and the other is near 800 nm. The first order derivative curves of reflectance spectral indicate that the first reflectance peaks shift to long wavelength according to the increased SSC. The ANN models make full use of the spectral information in different channels which can simulate the pigment and non pigment particles concentrations at same time. The fitting effects are preferable with R2 more than 0.95 for each model. The ANN mode can be used for satellite remote sensing inversion, especially MERIS data, because of its comparatively high spatial resolution. PMID- 19186798 TI - [Distribution form of nutrients in roof runoff]. AB - 6 rainfall-runoff events were examined from a concrete roof by a traffic artery in Shanghai to characterize the particle-bound and dissolved nutrients in roof runoff and to get more knowledge about roof runoff pollutants and lay the theoretic foundation for the selection of roof runoff purification method. Results indicated that event mean concentration (EMC) of TN ranges between 4.208 mg/L and 8.427 mg/L compared to 0.078-0.185 mg/L for TP. DN and PP are respectively the primary forms of TN and TP. During the runoff, the number ratio of small particles is gradually increased. The dynamic behavior of TP is similar to TSS, but the TN-TSS relationship in the course of runoff is not obvious. The increase of both PN/TSS and the number ration of small particles showed that nitrogen (microgramme per gram particle weight) attached to small particles is more than large particles. Regression analysis between TSS and TP & PP of samples in the early 10 min of runoff results in a high R2, but the relationship between TSS and TP & PP of samples in the entire runoff is not as close as the early 10 min, which reflects that the phosphorus quantity attached to the particle changes from the perspective of the entire course of runoff. First flush of TP is stronger than TN because the load discharge of dissolved nutrients is more stable than particle nutrients. In addition, more nitrogen absorbed by the unit mass small particles to a certain degree weakens the first flush of TN. PMID- 19186797 TI - [Combining ultrafiltration, fluorescence spectroscopy and HPSEC to characterize dissolved organic matter in surface waters]. AB - The combination of ultrafiltration, three-dimensional excitation/emission matrix (3DEEM) fluorescence spectroscopy and high-performance size-exclusion chromatography (HPSEC) was used to characterize the molecular weight distribution properties of different fluorescence materials, further revealing the differences in the sources and components. Dissolved organic matter (DOM) from different lakes fractionated by ultrafiltration with a nominal 1,000 molecular weight cut off regenerated cellulose membrane, then 3DEEM spectrophotometry and HPSEC were applied to investigate the characteristics of high molecular weight materials in retentate and low molecular weight materials in permeate. The result indicated that the flulvic acid-like (Ex/Em approximately equal to 260 nm/450 nm of peak A and Ex/Em approximately equal to 320 nm/439 nm of peak C) and protein-like (Ex/Em approximately equal to 275 nm/312 nm of peak B and Ex/Em approximately equal to 220 nm/308 nm of peak D) fluorophores in permeate were detected after ultrafiltration which is covered by high absorbing peak of humic-like (Ex/Em approximately equal to 360 nm/462 nm of peak E) fluorescence fluorophores in the original Shennonajia bog water, Hubei province. They permeated the membrane because of their low molecular weight distribution properties. The content of autochthonous protein-like (Ex/Em approximately equal to 280 nm/334 nm of peak B and Ex/Em approximately equal to 225 nm/328 nm of peak D) component was too low to be measured by high-sensitivity fluorescence spectrophotometer in the original water of Lake Hongfeng, Guizhou province. But they can be concentrated attribute to their high molecular weight distribution properties. The 3DEEM fluorescence spectroscopy of retentate exhibited evident protein-like fluorophores. Moreover, there had obviously difference in molecular weight between different sources of fulvic acid-like and protein-like components. It has been shown that the molecular weight distribution of autochthonous protein-like matter in Hongfeng lake is larger than allochthonous protein-like matter in Aha lake, Guizhou province. One was retained by membrane and another was permeated. Therefore, these techniques together will offer direct and convenient qualitative information about DOM in lake waters. PMID- 19186799 TI - [Residence time distributions and spatial variation of N, P in the subsurface flow constructed wetlands for purification of eutrophic aquaculture water]. AB - Hydraulic residence time distributions (RTD) and spatial variations of N, P were studied in a small-scale horizontal subsurface-flow constructed wetlands (HSFCWs) planted with Cyperous alternifolius and Typha angustifolia respectively for purification of eutrophic aquaculture water. The results show that the residence time distribution curves of the investigated HSFCWs lie between plug-flow and completely mixed model with characteristic values (sigma2) of 0.3246 and 0.4108, respectively. Compared with Typha angustifolia, Cyperous alternifolius wetland shows fine flow pattern with characteristics of smoother RTD curve and weaker vertical mixed flow. Total nitrogen (TN) and ammonia nitrogen (NH4+-N) show stratified distributions in the two HSFCWs, especially in the front end of the wetland beds. TN in the lower layer is higher than that in the upper, while NH4+ N in the middle layer is the lowest in all the sampling layers. Total phosphorus (TP) and phosphate (PO4(3-)-P) increases with sampling depth. Differences in TP and PO4(3-)-P between the layers decrease gradually along distance. Cyperous alternifolius wetland shows better stratification distributions of N, P, as compared with Typha angustifolia, which is mainly contributed to the difference in flow patterns between the two HSFCWs. On average, concentrations of TN and TP in the rear end of the Cyperous alternifolius wetland are 19.3% and 12.5% lower, respectively, as compared to the Typha angustifolia wetland, suggesting that removal efficiencies of the Cyperous alternifolius wetland for purification of eutrophic aquaculture water is higher than those of the Typha angustifolia. PMID- 19186800 TI - [Removal technology of typical odorant in drinking water]. AB - To eliminate the odor in drinking water of one City in China, a study was performed on the typical odorant removal technology and removal processes. Its results showed that as typical odorants, ethanethiol was effectively removed by oxidation and geosmin and 2-MIB were removed by both oxidation and adsorption, but geosmin and 2-MIB's adsorption effect was better than oxidation. When thiol and thioether in raw water was less than 20 microg/L, furthermore, there was not any other odorant, potassium permanganate oxidation should be equipped with enhanced coagulations. When geosmin and 2-MIB in raw was less than 30 ng/L, activated carbon adsorption should be equipped with enhanced coagulations. When thiol and thioether in raw water was more than 20 microg/L, or geosmin and 2-MIB was more than 30 ng/L, ozone-activated carbon process should be added after the conventional water treatment process. When thiol and thioether in raw water was more than 150 microg/L, or geosmin and 2-MIB was more than 100 ng/L, preoxidation or adsorption of powder activated carbon at intake should be combined with ozone activated carbon according to the odorant composing. PMID- 19186801 TI - [Reaction of free chlorine transforms into chloramines in microorganism inactivation experiment]. AB - Due to organic N exiting in the pure bacterial cultures, when using free chlorine to inactivate microorganisms, freest chlorine transforms into organic N chloramines which are no use in disinfection process. This reaction happens in E. coli, Pseudomonas aeruginosa inactivation experiments. Added 2 mg x L(-1) free chlorine to inactivate E. coli of 10(8) CFU x mL(-1), free chlorine exhausts in 5 min while chloramines and dichloramines become 0.92 mg x L(-1) and 0.4 mg x L( 1). E. coli is used as test microorganism in following inactivation experiments. E. coli suspension is purified by centrifugal and membrane filtration, but the purification can't help to separate the organic N from E. coli suspension, which indicating that the source of organic N may be from bacterium cells. Dilution is worked when E. coli concentrations are diluted under 10(5) CFU/mL. This result should be regarded in related researches in future. PMID- 19186802 TI - [Sorption-desorption of phosphate in wastewater by hydrous iron oxide]. AB - FeCl3 was used t o prepare hydrous iron oxide (HIO) as a n absorbent for phosphate (P) sorption and desorption study. The results showed that as pH decreased, the sorption capacity of HIO increased, and the sorption kinetics followed the second-order model, and the sorption isotherm could be fitted by the Langmuir equation. A 50 g/L NaOH solution was used for desorption of P from HIO, and the desorption rate could be reached over 98% . No relation was found between desorption rate and adsorption capacity. Based on above results, HIO was applied to adsorption of P from supernatant of sludge thickener, and after desorption, more than 90% of P was recovered. According to the results obtained, an effective system for P removal and recovery from municipal wastewater was suggested, which includes the following processes: adsorption, desorption, regeneration of HIO, and of recovery of P from P-rich desorption solution. PMID- 19186803 TI - [Coagulation performance and mechanism of Alb species in polyaluminum chloride in removing humic acid]. AB - Alb species [Al12 AlO4 (OH)24(7+)] is multinuclear hydroxyl compound formed in PAC hydrolysis-polymerization process, with properties of small particle sizes, much positive charge, high aggregation degree and large molecular weight. Alb species was purified from PAC containing high concentration of Alb species using ethanol-acetone mixed precipitation method. The influence of dosage, pH and inorganic ions on humic acid removal from simulative water was studied by measuring zeta potential and UV254. The PDA output (FI curve) was analyzed and the relationship between the parameters (S1, S2, H1, H2) of FI curve and coagulation mechanism were studied at different Alb dosage and pH. When the dosage was 4.3 to 6.3 mg x L(-1) and pH was 3.0 to 6.0, the highest humic acid removal efficiency was obtained. The existence of NH4+, SiO3(2-) and H2PO(-) inhibits the coagulation performance. The results of coagulation kinetics and Zeta potential studies revealed that Alb acted mainly by charge neutralization as well as adsorption bridge building in water treatment. PMID- 19186804 TI - [Treatment of high algae-laden water by step-recycle counter current flotation]. AB - High algae-laden water was treated respectively by single-recycle and step recycle counter current flotation. The results indicate that step-recycle mode of dissolved air water can strengthen capturing of bubbles and flocs association suspension bed, and extend the collision time of smaller flee and bubble because of reducing the thickness of suspension bed and increasing that of transition bed. Given the condition that other parameters were same, the biggest hydraulic loading of single-recycle could reach 11 m/h, and that of step-recycle could reach 17 m/h which is increased by 50%. Efficiency of treatment was improved by a large margin. The recycle ratio of two releasers and position of the pensile releaser influenced removal effect remarkably. The distance between settled releaser and inlet of raw water is 180 cm, and the range of distance between pensile releaser and inlet of raw water is 60-90 cm. The flow of pensile releaser is 2-3 times as much as settled releaser. PMID- 19186805 TI - [Pilot-scale opposite folded plate hybrid anaerobic reactor (OFPHAR) in treatment of sewage]. AB - Based on the theories of mass-transfer and two-double integrated staged multi phase anaerobe (TSMPA), a pilot-scale opposite folded plate hybrid anaerobic reactor (OFPHAR) was designed to treat low concentration sewage. All the trial lasted 12 months and the results indicated that the optimal HRT was 6h. At this HRT, the COD, TP and TN removal rate were 78.58%, 35.15%, 39.17%, respectively, at 25 degrees C +/- 2 degrees C. The optimal rate of anaerobic section was 45% 65%. Controlled HRT = 6 h, the COD, TP and TN removal rate were 64.37%, 20.72%, 23.65%, respectively, and the specific methane production capacity were 1.85 mL/(g x h) when the temperature decreased to 7 degrees C. The results of trial indicated that apply this OFPHAR to treat low concentration sewage at low temperature in north China is feasible. PMID- 19186806 TI - [Experiment study on the metamorphic amylum production wastewater treatment by anaerobic baffled reactor]. AB - The present study reports the start-up of treating metamorphic amylum production wastewater by anaerobic baffled reactor (ABR) and the bio-chemical features of granular sludge. The optimal conditions in treatment of the metamorphic amylum production wastewater were proposed, and the running performance in treating salt containing wastewater was studied. Our results show that the common anaerobic activated sludge can be acclimated by increasing the organic loading and salt concentration at the same time, and the granular sludge tolerant to low salt can be achieved by the acclimation. When chloric ion was 8,500 mg/L and salinity was 1.6%, the anaerobic activated sludge could degrade organic materials normally in the wastewater after the acclimation, and the COD removal is over 85%. When the concentration of metamorphic amylum production wastewater in the experiment was 12,640 mg/L and the optimal hydraulic retention time was 48 h, the removal efficiency of COD was 85.9%. Effect of sharp decrease of chloric ion concentration on sludge microorganism is larger than that of the sharp increase in the system. The system can endure the change of chloric ion concentration by increasing from 8,500 mg/L to 12,500 mg/L or decreasing from 8,500 mg/L to 4,500 mg/L, and it is more tolerant to the sudden increase than that of the sudden decrease of chloric ion concentration. The ABR system can treat the wastewater with chloric ion below 15,000 mg/L and salinity of about 2.5%. PMID- 19186807 TI - [Micro-ecology mechanism on microaerobic treatment performance at various influent COD concentrations or HRTs]. AB - A lab-scale continuous experiment was designed to investigate the effect of influent COD concentrations (11,000, 15,000, 30,000 mg/L) or hydraulic retention time (HRT 42, 25 h) on the microareobic treatment performance of simulated high strength organic wastewater. The molecular microbiological technologies, including Fluorescent in-situ hybridization-Flow Cytometry (FISH-FCM), Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis (PCR-DGGE), and Biolog-FF assay method, were used to detect the variation of microbial community in the aerated column during the four pseudo-steady-state periods. The yeast contents remained > 99.9% throughout the overall experimental periods according to FISH FCM. Increasing influent COD concentration brought on a rising MISS (2.0-7.3 g/L) and a reduced specific COD removal rate [2.3-1.7 kg/(kg x d)], structural (PCR DGGE)/metabolic (Biolog FF) diversity index values of fungal community in the aerated column had an increase of (2.05-2.19)/(4.42-4.45). Shortening HRT brought on a reduced MISS (7.3-6.0 g/L) and a rising specific COD removal rate [1.7-2.8 kg/(kg x d)], structural (PCR-DGGE)/metabolic (Biolog FF) diversity index values of fungal community had a decrease of (2.19-0.79)/(4.45-4.36). Increasing influent COD concentration or shortening HRT has an absolutely adverse effect on the microaerobic treatment performance and micro-ecology in the aeration column although either of them can increase influent COD loading up to a higher level. PMID- 19186808 TI - [Relation analysis of phosphorus removal and BOD5 loading using PHB monitoring in A2/O process]. AB - In a pilot-scale anaerobic-anoxic-oxic experimental system for municipal wastewater treatment, PHB consumption in an oxic tank and PHB production in an anaerobic tank had been monitored overtime, and relations among PHB consumption/production, phosphorus removal/release and BOD5 loading had been analyzed. The results indicated that the consumption of PHB was positively related with phosphorus removal, and about 140 mg PHB consumption could result in 1 mg P removal. In the anaerobic tank, about 1.17 mg P release could result in 100 mg PHB production. The PHB production at the loading of 0.176 g/(g x d) was 4 mg/g MLSS less than that at 0.413 g/(g x d). No significant relevance was observed between PHB and BOD5 removal. The PHB of microorganism synthesizes was increased with the increase of temperature. Compared with the lowest temperature (17.1 degrees C), 20% of the total PHB content was increased at the maximum temperature (33.2 degrees C) in active sludge. PMID- 19186809 TI - [Experimental study on the effect of COD/P ratios and phosphate recovery on a BNR system]. AB - Based on a BNR system--BCFS, the effect of COD/P and phosphate recovery on the BCFS process was experimentally investigated. The results indicate that a higher P-removal efficiency (87%) and a good P-effluent quality (< or = 0.8 mg x L(-1)) could be maintained at COD/P ratios higher than 24. When COD/P was reduced below 24, the P-removal efficiency was suddenly decreased down to 62% and the P effluent went up to 5 mg x L(-1). Under the circumstance, chemical phosphate recovery in the side-stream of anaerobic supernatant was combined with bio-P removal. When the side-stream increased up to 30%, the P-effluent went back to < 1 mg x L(-1), with a corresponding phosphate recovery efficiency of 54%. PMID- 19186810 TI - [Experimental determination of bacterial decay characteristics in biological wastewater treatment system]. AB - The characteristics of cell decay in biological wastewater treatment systems were investigated under aerobic condition, by measuring the decay rate and by determining the death rate with LIVE/DEAD dyeing experiments. It was found that cell decay in biological wastewater treatment systems can be actually described as two parts: decay caused by cell death and decay derived from activity decrease. The experimental results revealed that 60% of cell decay in a nitrifying system was caused by activity decrease and 40% was caused by cell death. In a heterotrophic system, however, activity decrease was responsible for 80% of cell decay, and the other cell decay for 20% was caused by cell death. PMID- 19186811 TI - [In-situ determination of endogenous decay rate of heterotrophic biofilm by microelectrodes]. AB - A separated oxygen microelectrode was constructed, based on the improved structure, to determine the oxygen distribution in biofilm at an endogenous state. Based on the diffusion-reaction model, the endogenous decay rate was evaluated by curve-fitting with the measured oxygen profiles. The results showed that the fluctuation range of output signal of oxygen microelectrode was reduced from (1.64 +/- 0.25) nA to (1.53 +/- 0.06) nA, the signal became stable with the fluctuation rejection ratio of 19.41 dB after the structural optimization. The endogenous respiration rate and the endogenous decay rate of heterotrophic biofilm were evaluated to be 2.979 mg x h(-1) and 0.0059 h(-1). PMID- 19186812 TI - [Nucleation of flocculent sludge in anaerobic reactor]. AB - The nucleation of flocculent sludge as seed in a mesophilic Internal Circulation Anaerobic reactor was investigated with a quantitative method proposed by us in this paper. In the 85th days, the average sludge diameter increased from 47.8 microm to 96.1 microm, which indicated that the nucleation was completed. During this process, the average sludge diameter had a significant linear correlation with the time with coefficient of 0.9893, the increase rate of average sludge diameter was 0.58 microm/d. Nucleus ratio of sludge increased from 7.6% on the 1st day to 36.1% on the 85th day. The increase rate of the nucleus ratio fluctuated in a wide range, experienced 'fast-slow-fast' three stages. During the experiment, the concentration of extra cellular polymers (ECPs) of the sludge was significantly positively correlated with the increase rate of the nucleus ratio, which indicated that ECPs could affect nucleation rate. During nucleation, the bio-activity of the sludge increased, but the improvement of settlement ability was not significant. The above-mentioned quantitative method and results would be useful to understand the sludge granulation mechanism on the level of reactor. PMID- 19186813 TI - [Reactivation characteristics of stored aerobic nitrifying granules]. AB - A sequencing batch airlift reactor (SBAR) was conducted to study the performance of physical characteristics and respirometric activities during reactivation of aerobic granules after a 2-month storage time. Results showed that the color of the aerobic granules had completely become dark, the size and settleability did hardly change during the storage period. The color turned to be brownish yellowish when the reactor was restarted for 2 weeks. The MLSS, granule size, and settleability increased rapidly along with reactivation. The heterotrophic activity resumed 86% within 1 day and was fully reactivated after 5 days. Then the COD removal rate was stabilized above 80%. Higher aeration rate and longer cycle time facilitated the reactivation of nitrifiers. When aeration rate was increased from 0.05 m3 x h(-1) to 0.10 m3 x h(-1) at 41st day, activities of ammonia-oxidizing bacteria and nitrite-oxidizing bacteria were improved from 88% and 82% to 122% and 92%, respectively. The removal rate of ammonium-nitrogen promptly reached above 96%. Nitrite-oxidizing bacteria were fully reactivated after the cycle time was prolonged from 4 h to 6 h. PMID- 19186814 TI - [Biodegradability of extracellular polymeric substances (EPS) produced by aerobic granules]. AB - Aerobic granules were cultivated with synthetic wastewater and were added in the course of aerobic starvation. Results showed that EPS produced by aerobic granules was composed of 40% biodegradable and 60% nonbiodegradable EPS. Among these, only biodegradable EPS could be utilized by their producers, while nonbiodegradable EPS contributed to maintain the spatial structures of aerobic granules. EPS extracted from fresh aerobic granules was fed as the sole carbon source to their own producers and activated sludge and was utilized by them. The average biodegradation rate of activated sludge in terms of EPS was 1.5 times faster than that of aerobic granules. When EPS extracted from starved aerobic granules was fed as the sole carbon source to their own producers and activated sludge, it could not be utilized and could not be energy source. PMID- 19186815 TI - [Effect of temperature on performance of microbial fuel cell using beer wastewater]. AB - The effects of temperature on performance and biological community structure were investigated in air-cathode microbial fuel cells (MFCs) using beer wastewater amended with 50 mmol/L phosphate buffer solution (PBS). The maximum power density decreased from 483 mW/m2 to 435 mW/m2 when the temperature varied from 30 degrees C to 20 degrees C, meanwhile just a little decreasing on coulombic efficiency and the COD removal rate were observed. Decreasing of temperature resulted in effects both on cathode potential and anode potential, but cathode potential behaved much more sensitive to temperature. The half-saturation constants (Ks) obtained from the fit of Monod-type equation were 228 mg/L (30 degrees C) and 293 mg/L (20 degrees C) respectively. Denaturing gradient gel electrophoresis (DGGE) analysis indicated that operating temperature not only affected the predominant population of the anodic bacterial community, but also had a great impact on the diversity of the cathodic microbial population. PMID- 19186816 TI - [Application of biological aerated filter in treating biomass wastewater and its microbial population characteristics]. AB - Biomass wastewater contains plenty of cellulose, hemicellulose, starch, saccharide, organic acid and protein. It has high COD value and leads to environment pollution and resource waste. Biomass wastewater was treated in this investigate using a biological aerated filter (BAF) on laboratory scale, and the effect of treatment in different hydraulic retention time (HRT) had been studied. The results show that the optimum HRT is 10 h. The value of COD and TN in effluent is 74.90 mg/L and 1.21 mg/L respectively when run 10 h, which are under the highest concentration of the second category of pollutant discharge allowed in the state sewage discharge standard (GB 8978-1996). When the system is stabile, evaluating the population characteristics of dominating microorganism in BAF with biology analysis such as microbial screening, single bacteria evaluation and 16S rDNA sequences analysis. The dominant bacteria belonged to Pseudomonas sp., Acinetobacer sp., Stenotrophomonas sp., Rhodococcus sp.and Microbacterium sp.. PMID- 19186817 TI - [Radiolysis of aqueous thiocyanate and its complicated system]. AB - We studied the 60Co gamma-ray radiolysis of thiocyanate aqueous solutions in the presence of free radical scavengers, determination of cyanide of radiated thiocyanate solutions, degradation of cyanide and thiocyanate complicated system and cyanide, thiocyanate and copper (I) cyanide complicated system. The presence of NaHCO3 and n-butanol made the degradation efficiency of thiocyanate decreased, in the radiation procession of thiocyanate, cyanide was produced; and the possible reaction equation is SCN(-) + 8*OH-->SO4(2-) + CN(-) +4H2O, in the cyanide and thiocyanate system, the thiocyanate degradation rate can obtained 30%, free cyanide degradation was little effected by coexistence of thiocyanate; and due to strong complex ability of copper (I), the main form of copper (I) cyanide is Cu(CN)3(2-), the total cyanide of the system can't be degraded completely and degradation rate is about 80%. PMID- 19186818 TI - [Growth of Microcystis aeruginosa affected by allelochemicals of Arundo donax Linn. extracted with different solvents]. AB - To study the effects of allelochemicals of Arundo donax Linn. on the growth of Microcystis aeruginosa, the allelochemicals were extracted with three solvents (methanol, ethyl acetate and hexane), respectively. Based on the observation of algal morphology and the measurement of algal density and cell size, the results showed the allelochemicals extracted with all the three solvents had inhibition on M. aeruginosa. The appearance time of allelochemical inhibition increased as follows, allelochemicals extracted with methanol < ethyl acetate < hexane. As treatment time extended, M. aeruginosa regrew at low doses of allelochemicals extracted with methanol. Their inhibitory effects at high doses were lower than those extracted with ethyl acetate and hexane. After 2 d and 4 d of treatment, the inhibition ratios of allelochemicals extracted with ethyl acetate and hexane reached almost 100%, respectively. The 50% effective concentrations (EC50, 6 d) of allelochemicals extracted with methanol, ethyl acetate and hexane were 0.17 g x L(-1), 0.05 g x L(-1) and 0.08 g x L(-1), respectively. The allelochemicals extracted with methanol caused cavities in cells, those with ethyl acetate caused cells into cavities, pieces and conglomeration, and those with hexane caused goffers on cells initially with conglomeration later. The allelochemicals extracted with all the three solvents decreased the cell size of M. aeruginosa, among which those extracted with ethyl acetate had the strongest action. PMID- 19186819 TI - [Acute toxicity effects of three red tide algae on Brachionus plicatilis]. AB - Acute toxicity testing method was used to study effects of different density of Prorocentrum donghaiense, Heterosigma akashiwo and Alexandrium tamarense on mortality rates and population growth parameter of Brachionus plicatilis under controlled experimental conditions. Results showed that 24 h LC50 values of Prorocentrum donghaiense, Heterosigma akashiwo and Alexandrium tamarense treatment to mortality rate of Brachionus plicatilis were 3.56, 1.21 and 0.49 (x 10(4) cells/mL) respectively. Marked density effects were presented when three species of red tide microalga showed their toxicity to Brachionus plicatilis. There were significant inhibitory effects on Brachionus plicatilis when it was exposed to cells of Prorocentrum donghaiense at the concentration of 10(4) cells/mL, filtrate and cell contents of Heterosigma akashiwo at the concentration of 10(5) cells/mL, and cells, filtrate and cell contents of Alexandrium tamarense at the concentration of 10(3) cells/mL respectively. Inhibitory effects of three species of microalga on Brachionus plicatilis were enhanced with increasing of microalgal density. PMID- 19186820 TI - [Gene cloning, sequence analysis and tissue expression of estrogen-related receptor alpha (Erralpha) in Japanese medaka and its transcriptional responses after differential EDCs exposure]. AB - Endocrine disrupting chemicals (EDCs) can bind or block nuclear receptors in the body and subsequently affect growth, development and reproduction of fish. Estrogen-related receptors (ERRs), belonging to the nuclear receptor superfamily, have been implicated in diverse physiological processes in estrogen signal pathway in mammals, while little is known about them in fishes. Complete mRNA sequence of ERRalpha from medaka (Oryzias latipes) was cloned, and the sequence is similar to those of other vertebrates, especially that the DNA-binding domain (DBD) of ERRalpha is highly conserved among the vertebrates (97.4%-100% sequence identities) and the ligand-binding domain (LBD) of medaka ERRalpha is 66.4%-67.0% sequence identities with those of mammals. The DBD of medaka ERRalpha is of the same length and has high sequence identity with those of estrogen receptor (ERalpha and ERbeta) and androgen receptor (ARalpha and ARbeta) of medaka, but much difference was found between the LBD of medaka ERRalpha with those of ERalpha, ERbeta, ARalpha and ARbeta. ERRalpha gene is located in chromosome 14 and is consisted of 5 exons. The expressions of ERRalpha in different tissues and the transcriptional responses of ERRalpha in testis of medaka exposed differential EDCs were studied by quantitative real-time RT-PCR. ERRalpha is expressed at apparently high levels in gonad, brain, eye, spleen and intestine, though it was broadly expressed in tissues. Significant transcriptional difference was found between testis and ovary, implying ERRalpha would be involved in sex differentiation and gonad development in fish. After 3 weeks exposure of medaka to 200 ng/L ethynylestradiol (EE2), 200 ng/L estrone (E1), 200 ng/L diethylstilbestrol (DES), 100 microg/L atrazine (AT) and 200 ng/L 17beta estradiol (E2), transcripts of ERRalpha were significantly decreased to 0.54, 0.56, 0.61, 0.63 and 0.65 of control (p < 0.05) in the testes, respectively. And those in the 1 microg/L tributyltin (TBT) and 1 microg/L triphenyltin (TPT) exposure groups were up-regulated to 1.34 and 1.35 folds of control (p > 0.05), respectively. These results suggested that ERRalpha would take actions in the disruption of sex differentiation and gonad development in fish by EDCs. In addition, no multiple steroid hormone-response element half-sites was found in medaka, which were reported in the upstream of ERRalpha gene in mammals, indicating there would be different regulation patters of ERRalpha between teleost and mammal. PMID- 19186821 TI - [Construction of recombinate luminescence bacteria vector to evaluate the genetoxic of environment pollutant]. AB - Recombinate luminescence bacteria have the important role in evaluating water toxicity. Two recombinate luminescence bacteria vectors PUCD-uvrA and PUCD-alkA were constructed to investigate the impaired mechanism of pollutant genetic toxicity. The genes of uvrA and alkA were amplified by PCR from E. coli W3110, sequenced after ligated with pGEM-T easy vector. The PCR products and PUCD615 vector were all digested with BamH I, EcoR I, then be linked and imported into JM109 with electrotransformation. Several clones were selected and identificated by PCR and sequencing. The results reviewed that the length of the uvrA and the alkA fragments were 237 bp, 326 bp. When they were sequenced and blasted in GenBank, the homology of sequences reached 99% indicated the amplified results correct. The results of sequencing ligated with PUCD615 reviewed that the fragments of uvrA and alkA had been inserted into the multiple clone site correctly, the insert direction and reading frame were also exactly. Optimizing the condition of ligation and transformation, the large fragment of PUCD615 and the short inserted sequences can been ligated successfully. PMID- 19186822 TI - [Functional correlation of dibenzothiophene and benzothiophene desulfurization enzymes]. AB - Gordonia sp. C-6 can desulfurize benzothiophene (BT) as the pathway similar to the "4S" pathway of dibenzothiophene (DBT)-desulfurizing, but the strain can not grow with DBT as the sole sulfur source. At current, there were not related reports on BT-desulfurizing genes at home or abroad. The DBT-desulfurizing genes of Rhodocossus erythropolis DS-3, dszA, dszB, dszC and dszABC were introduced into Gordonia sp. C-6 respectively using a Rhodococcus-E. coli shuttle vector, to construct new recombinant strains Gordonia sp. CRA, Gordonia sp. CRB, Gordonia sp. CRC and Gordonia sp. CRABC, the enzyme activities of which was respectively 76.8 micromol x (g x h)(-1), 51.6 micromol x (g x h)(-1) and 62.4 micromol x (g x h)(-1), increasing by 1.5 times compared with 35.2 micromol x (g x h)(-1), 21.3 micromol x (g x h)(-1) and 25.5 micromol x (g x h)(-1) of wild strain Rhodocossus erythropolis DS-3. Of the recombinant strains, only recombinant strains Gordonia sp. CRC with DszC and Gordonia sp. CRABC with DszABC exhibite significant growth with DBT as the sole sulfur source, Gordonia sp. CRA with DszA and Gordonia sp. CRB with DszB could not live with DBT as the sole sulfur source. The results show that, DBT monoxygenase and BT monoxygenase, catalyzed the first two steps of DBT and BT oxidation, respectively, are the key enzymes responsible for substrate recognition, however, the enzymes catalyzed the last two steps have the similar substrate specificity. The active sites of the two monoxygenase could be predicted by comparing with the sequence differences of their amino acids. PMID- 19186823 TI - [Screening of two phenanthrene-utilizing and high-effective phosphorus accumulating bacteria and their effects on phosphorus-accumulating characteristics]. AB - In this study, two phenanthrene-utilizing and high-effective P-accumulating bacteria Y11 and Y4-2 were isolated from sludge samples of Taihu Lake using plate culture and blue-colored colonies methods. Strains Y11 and Y4-2 were identified as Acinetobacter sp. according to the results of morphology, physiology and the phylogenetical analyses of 16S rDNA sequences. The two strains showed high effective P-accumulating ability in plate and broth cultures. Y11 could grow well from 10 degrees C to 35 degrees C and pH from 6 to 9, Y4-2 could grow well from 10 degrees C to 35 degrees C and pH from 6 to 8. Phenanthrene could be used as sole carbon and energy sources. In our experiment, high phosphorus concentration in broth culture had no negative effect on the growth of strain Y11, however the growth of strain Y4-2 was slightly affected. Under cultivation condition of 30 degrees C, 170 r/min, 1% inoculation (D600 = 0.4), the accumulation of phosphorus was various by strains and cultivation concentration of phosphorus as well: In 2 mg/L phosphorus concentration medium, the highest phosphorus-accumulation ratio of strains Y11 and Y4-2 were 96.13% and 94.65%, the phosphorus concentration decreased from 2 mg/L to 0.08 mg/L and 0.11 mg/L, respectively. In 5 mg/L phosphorus concentration medium, the highest phosphorus-accumulation ratio of strains Y11 and Y4-2 were 95.94% and 71.19%, the phosphorus concentration decreased from 5 mg/L to 0.20 mg/L and 1.44 mg/L, respectively. In 8 mg/L phosphorus concentration medium, the highest phosphorus-accumulation ratio of strains Y11 and Y4-2 were 71.24% and 47.81%, the phosphorus concentration decreased from 8 mg/L to 2.30 mg/L and 4.18 mg/L, respectively. For the P removal of Yunnan Dianchi water sample containing 1.01 mg/L phosphorus, Y11 and Y4-2 were successful to decrease the phosphorus concentration from 1.01 mg/L to 0.06 mg/L in 6 h and 48 h, respectively. The results indicated that Acinetobacter sp.Y11 and Y4-2 possess high ability of phosphorus removal and suitability to various water environments. Y11 strain adapt to all kinds of eutrophic waterbody even polluted with phenanthrene, Y4-2 adapt to pH < or = 8.0 eutrophic waterbody with phenanthrene pollution. PMID- 19186824 TI - [Aerobic microbial degradation of polybrominated diphenyl ethers]. AB - The biodegradation of 4, 4'-dibromodipheny ether (BDE15) and decabromodiphenyl ether (BDE209) by white rot fungi under aerobic conditions was studied. Effects of non-ionic surfactant Tween 80 and beta-cyclodextrin as solubilizers on the apparent solubilities and biodegradation rates of BDE15 and BDE209 were also evaluated. The results showed that both BDE15 and BDE209 were efficiently degraded by white rot fungi. The degradation rates were 43.0% and 62.5% for BDE209 and BDE15, respectively, after 10 d incubation. The degradation of BDE209 was greatly enhanced by addition of Tween 80 (< or = 700 mg/L) and beta cyclodextrin, which may own to their solubilization effects on BDE209. However, Tween 80 at a high concentration (900 mg/L) would restrain the fungal growth, thereby decrease the degradation of BDE209. Addition of Tween 80 and beta cyclodextrin exhibited some negative effects on the degradation of BDE15, which may due to decreased concentration of free BDE15 in water solution resulted from inclusion function of Tween 80 micelles and beta-cyclodextrin cavity, although the apparent solubility of BDE15 was drastically increased by both of them. PMID- 19186825 TI - [Characteristics of 4-chlorophenol degradation by a soil bacterium Acinetobacter sp]. AB - The bacterium capable of degrading 4-chlorophenol (4-CP) was enriched and isolated from agricultural soil. It was gram-negative, rod-shaped bacterium, and identified as Acinetobacter sp. based on the analysis of the 16S rRNA gene fragment. It was able to utilize 4-CP as a sole carbon source. The degradation mechanism of 4-CP by this isolate was proposed as a modified ortho-cleavage pathway, the activity of chlorocatechol 1,2-dioxygenase was markedly induced. The bacterial isolate grew well and exhibited a high degradation efficiency when the initial concentration of 4-CP was between 2-8 mmol/L. It was able to survive in the presence of 4-CP at higher concentrations (up to 8 mmol/L). Not only 4-CP, but also 2-chlorophenol, 3-chlorophenol, phenol, and 2,4-dichlorophenol, were also growth substrates for the isolate. The results of co-substrate supplementation illustrated the suitable conditions of the isolate to improve growth rate and 4-chlorophenol biodegradation efficiency. The results suggested that the isolate had a potential use for bioremediation of the site contaminated with 4-chlorophenol. PMID- 19186826 TI - [Selection and kinetic mechanism of psychrotrophs in low temperature wastewater treatment]. AB - A study was carried out taking low temperature domestic wastewater as target in laboratory, the composition of bacteria in activated sludge was analyzed and the degradation kinetics of organics was investigated. Six preponderant psychrotrophs were selected out, belonging to Zoogloea, Aeromonas, Flavobacterium, Micrococcus, Bacillus and Pseudomonaus, respectively. Results show that TTC-dehydrogenase activity of the psychrotroph is 25.44 mg/(L x h) which is 20.5 times more than that of ordinary activated sludge at 4 degrees C. The efficiency of COD biodegradation by psychrotrophs at low and normal temperature is 80.9% and 73.4%, respectively. Mesophilic bacteria almost lost their activity at low temperature. Kinetic analysis shows that biodegradation of organics by psychrotrophs at low and mesothermal temperature as well as mesophilic bacteria at mesothermal temperature are in accordance with the model of first-order reaction. Psychrotrophs, which assure the removal efficiency of organic pollutants at low and normal temperature, could keep the uniform reaction velocity as the mesophilic bacteria and also adapt wide ecological amplitude of temperature. Efficient psychrotrophs were immobilized on soft polyurethane foams which acted as carriers in the experiment at 4 degrees C, as a result of that, the removal efficiency of COD was increased about 18% higher than that of mobilized ones at low temperature, the biodegradation kinetics of COD by immobilized psychrotrophs also followed the first-order reaction model. With glucose in water as the source of nutriment, the reaction velocity of immobilized bacteria is 2.35 times higher than that of the mobilized ones. By the immobilized psychrotrophs biodegradation of varied nutriment, the effluent could achieve the first-degree B of pollutants discharge standard for municipal wastewater treatment plant. PMID- 19186827 TI - [Stimulation of biodecolorization of direct yellow 11 by decolorization products of reactive black 5]. AB - This study focused on the effects of decolorization products of reactive black 5 (RB5) on anaerobic decolorization of direct yellow 11 (DY11) by Rhodopseudomonas palustris W1 and its reaction mechanisms. The results showed that the decolorization products of RB5 could obviously accelerate biological decolorization rate of DY11. For initial concentration of 200 mg/L of DY11, the addition of decolorization products of RB5 resulted in that the decolorization kinetic constant K was increased from 17 mg/(L x h) to 42.5 mg/(L x h), followed by an further increase to 48.8 mg/(L x h) after optimization of RB5 supplement. Cyclic voltammogram analysis demonstrated that the decolorization products of RB5 presented electrochemical activity due to the presence of redox electron transfer mediators, as indicated by an observation of two current peaks of reversible oxidation and reduction appeared at potential of ca. 83 mV and -200 mV, respectively. According to LC-MS and FT-IR analysis, the electrochemically activated species of the decolorization products were characterized to be 7-amino 8-hydroxy-1, 2-naphthaquinone-3, 6-disulphonate-1, 2-diimine (TAHNDSDP-1). TAHNDSDP-1 with quinine-like structure was able to transfer between its oxidative and reductive forms by transferring 2[H], which was supposed to be the redox mediator to shuttle electrons from electron donors to azo dyes, thus, improving the reduction of de materials. PMID- 19186828 TI - [Characteristics of Ag+ biosorption by the waste biomass of Saccharomyces cerevisiae]. AB - To understand the characteristics of Ag+ biosorption by the waste biomass of Saccharomyces cerevisiae, the effect of environmental factors on metal biosorption were studied, including sorption time, initial concentration of metal ions, temperature, initial pH value, ionic strength. The relevant kinetics, equilibrium and thermodynamics were discussed. The results showed that biosorption of Ag+ on the biomass were very rapid processes. Ag+ uptake quantity within 10 min arrived at 86% of that of the equilibrium uptake during 24 h uptake when initial concentration of Ag+ was 1 mmol x L(-1) and the biomass concentration was 2 g x L(-1) x Ag+ uptake process followed the pseudo-first and the pseudo-second order kinetics. The pseudo-second order kinetic gave better fitting results than the pseudo-first order kinetic. The equilibrium data could be fitted well with the Langmuir model, better than the Freundlich model. The maximum sorption capacity obtained from the Langmuir model was 0.385 mmol x g(-1) when initial Ag+ concentration was 0-8 mmol x L(-1) and the biomass concentration of 2 g x L(-1). The values of the thermodynamic constants of biosorption were calculated and demonstrated that the biosorption process of Ag+ was spontaneous and the entropy increased. The biosorption process by the waste yeast was favorable for the removal and recovery of Ag+ from wastewater. PMID- 19186829 TI - [Ecological risk assessment of sediment pollution based on triangular fuzzy number]. AB - Based on the characteristics of random and fuzziness, and the shortage and imprecision of datum information of water environmental system, environment background value of sediments and concentration of pollution is calculated by means of triangle fuzzy number and fuzzy risk assessment model of the potential ecological risk index is established. Using this method heavy metal pollution and ecological risk in the Yangtze Estuary and its adjacent waters were analyzed. The result shows that the environment of the foundation of the study area is subject to varying degrees of pollution. The pollution extents are correspondingly Cu, Hg, Zn, Pb, As, Cd. RI by that method and the Hakanson ecological risk method is in similar trend. RI of the estuary, turbidity maximum zone and Hangzhou bay is greater than that at outside of the estuary and sea area nearby Zhousan, and the potential ecological risk rate increases one. The assessment result is good in the validation based on the corresponding period macrobenthic community parameters. PMID- 19186830 TI - [Aging characteristics of copper and zinc added to typical soils of China]. AB - The aging processes of copper and zinc in single and combined metal-contaminated typical soils of China, red soil, paddy soil and cinnamon soil, were studied. The results showed that available copper and zinc (metals extracted by 0.01 mol x L( 1) CaCl2 ) decreased rapidly at initial stages, and then reduced slowly, where the turning point occurred at about 90 d. The difference in aging of copper and zinc was insignificant whether in single metal-contaminated soils or in combined metal-contaminated soils, suggested there were similar chemical behaviors between copper and zinc. The aging processes of copper and zinc in red soil, paddy soil and cinnamon soil fitted best a second-order equation (R2 = 0.9940-0.9999, p < 0.0001), whereas parabolic diffusion equation has less goodness of fit. It indicated that the transformation from availability to unavailability of metals, i.e. aging, was not completely controlled by diffusion, but controlled by the interactions from surface nucleation/precipitation, occlusion by organic matter, and diffusion, etc. The aging of copper and zinc in soils was significantly affected by pH. In soils with low pH such as red soil, the ratio of available metals was higher and the aging rate was slower [constant of rate, k2 4.36x10(-3) 7.05x10(-3) kg x (mg x d)(-1)]; whereas in soils with high pH, for example in cinnamon soil, the ratio of available metals was lower and the aging rate was faster [k2 1.095x10(-2)-1.377x10(-2) kg x (mg x d)(-1)]. That is, the aging rate of metals in soils increased obviously with increasing pH. PMID- 19186831 TI - [TiO2 photocatalyzed As (III) oxidation and adsorption in variable charge soil systems]. AB - Toxicity and mobility of As(III) in environments is greater than As(V). The transformation of As(III) to As(V) is in favor of the adsorption and fixation of arsenic on adsorbents. TiO2 photocatalyzed As(III) oxidation in the suspension systems of variable charge soils and the adsorption of oxidation products by the soils were investigated with a batch method. The photocatalysis experiments were conducted in a reaction box made by authors. The experiment results show that the photocatalyzed As(III) oxidation increased with the increase in the amount of TiO2 added and the time of irradiation, and 1.0 g x L(-1) of TiO2 and 90 min of irradiation were enough for photocatalyzed As(III) oxidation in the soil suspension systems. The TiO2 photocatalyzed As(III) oxidation in the variable charge soils and the adsorption of oxidation products by the soils increased the removal of arsenic from water and the similar processes were observed in TiO2O Fe/Al oxides' systems. The results also indicated that gamma-Al2O3 was able to induce the photocatalyzed oxidation of As(III) in suspension systems. PMID- 19186832 TI - [Effects of phosphorus application on translocation of lead, zinc and cadmium in the soil-plant system]. AB - A pot experiment was conducted to determine the influence of phosphorus (P) application on the translocation and accumulation of lead (Pb), zinc (Zn), and cadmium (Cd) in a soil solid-liquid-plant system using a plant Swamp Morningglory (Ipomoea aquatica Forak) as an indicator and Visual MINTEQ model. A contaminated soil containing 22,830 mg x kg(-1) Pb, 746 mg x kg(-1) Zn, and 10 mg x kg(-1) Cd was in the root was significantly decreased after P application, while the Zn concentration were increased, but no impact on Cd concentration after P application, compared to the control. Further more, concentrations of Pb, Zn, and Cd in plant tissue were sharply reduced after P application by 53%-92%, 35%-71%, and 59%-86%, respectively; Concentrations decreased exponentially with increasing amount of P; Relationship between Pb and Zn (r = 0.993, p < 0.01), between Pb and Cd (r = 0.986, p < 0.01) suggested that there was a positive relationship between the mechanisms controlling the phytoavailability of Pb, Zn, and Cd. It was suggested that Cd concentrations in the soil solution were reduced generally by P application as a result of the increased soil pH, and Zn concentrations were no change, but Pb concentrations were increased. Species distribution showed that PbHPO4, PbOH+ and PbH2PO4+ were the main complexation species of Pb in the soil equilibrium solution after P application. It was also suggested that Pb accumulation by root were negatively correlated to Pb in soil solution with coefficient of -0.872 (p < 0.05), and that water soluble Zn and Cd concentrations were reduced while the concentrations in root tissues were increased after P addition. These results indicate the difference between Zn, Cd and Pb and suggest that P as KH2PO4 successfully decreased the phytotoxicity of Pb, Zn, and Cd in the contaminated soil. PMID- 19186833 TI - [Influences of fertilization and seasonal variation on microbial community in a Chinese mollisol]. AB - Fertilization and seasonal variation play very important roles in affecting microbial structure and activity, as a result, leading to the significant evolution of soil fertility. The effect of manure (MCK) and combined application of chemical fertilizers (NPK) on soil microbial biomass and structure were studied by measuring soil microbial biomass carbon (nitrogen) and phospholipid fatty acid (PLFA) in different microbial communities, with the nil-fertilization (CK) and fallow as controls. Results show the manure application significantly improves the soil nutrient contents and the amounts of Cmic and PLFA of different microbial communities. The amounts of fungal PLFA (8.40 nmol x g(-1)) and Cmic (322.5 mg x kg(-1)) and Nmic (57.9 mg x kg(-1)) are significantly higher than those of CK (5.4 nmo x g(-1), 152.6 mg x kg(-1), 32.1 mg x kg(-1), respectively) or NPK (3.5 nmol x g(-1), 144.3 mg x kg(-1), 30.7 mg x kg(-1), respectively). And the contents of Cmic, Nmic and PLFA of different microbial groups in NPK are lower than those in CK. Correlation analyses show the soil nutrient contents are significantly positively correlated with Cmic, different microbial PLFA contents and G(-)/total bacteria ratios, while negatively correlated with C+/G(-) bacteria ratio (p < 0.05). The principle component analysis of PLFA shows the microbial structures in different treatments and sampling dates are significantly different. Seasonal changes are also found to cause great fluctuations in soil basic properties, and microbial community structure in arable soils and fallow respectively cluster strictly together by sampling dates. The amount of Cmic is highest on April 11 (295.6 mg x kg(-1)), while Nmic (49.3 mg x kg(-1)) and PLFA contents are highest in summer (July-August); the lowest amounts of Cmic (184.2 mg x kg(-1)), Nmic (30.63 mg x kg(-1)) and PLFA exist on May 31. Fertilization and seasonal variations significantly affect soil fertility, microbial structure and activity. PMID- 19186834 TI - [Characters of soil-vegetable transfer and accumulation of polycyclic aromatic hydrocarbons]. AB - The transfer and accumulation of polycyclic aromatic hydrocarbons (PAHs) from aged contaminated agricultural soil to vegetable was studied. The results show that the positive correlation between the PAHs concentrations in tested vegetables and those in the corresponding soils is present. The PAHs concentrations in contaminated soils are significantly (p < 0.05) higher than that in roots, stems and leaves of vegetables, and the PAHs concentrations in roots are also significantly (p < 0.05) higher than that in stems and leaves. The ratio of sum of low molecular weight PAHs to total PAHs concentrations (Sigma LMW PAHs/Sigma PAHs) in polluted vegetable roots is higher than that in corresponding soils (p < 0.05), and LMW-PAHs are more easily accumulated in vegetable roots than HMW-PAHs (high molecular weitht PAHs). The distinction of PAHs concentrations among the four selected leafy vegetables in the same tested soils does not achieve the significant level. PMID- 19186836 TI - [Effect of temperature on methanogenic pathway during household waste anaerobic digestion by stable carbon isotopic signature of CH4]. AB - The methanogenic pathway during anaerobic digestion of household waste was investigated by stable carbon isotopic signature analysis, and testified by the analysis of gas production, leachate characteristics and microbial fluorescence in situ hybridization (FISH) methods. Furthemore, the difference of methanogenic pathway between mesophilic and thermophilic anaerobic digestion was also discussed. Results showed that under mesophilic conditions, the isotopic stable carbon signature of CH4 (delta13 CH4) initially decreased to -69.5 per thousand, indicating that CH4 was produced from CO2 and H2 by hydrogenotrophic methanogens. When active CH4 production phase started, the delta13 CH4 values quickly increased to -23.8 per thousand, which indicated more and more CH4 were formed by aceticlastic methanogens, dominantly the family of Methanosarcinaceae, shown by the FISH results. The delta13 CH4 values decreased successively and ultimately remained at -55 per thousand, indicating that the fraction of aceticlastic methanogenesis finally decreased to a steady level comparative with CO2-derived methanogenesis at the steady slow methane production phase. Under thermophilic conditions, the delta13 CH4 values remained at a level about -70 per thousand, showing that methane were solely produced from CO2 reduction, and acetate syntrophic oxidation happened during the active methane production phase. PMID- 19186835 TI - [Evaluation of soil lead contamination by biomarkers in leaves of Vicia faba L. seedlings]. AB - Stress proteins (HSP70 and HSP60) and other related biological parameters were investigated in leaves of Vicia faba L. seedlings subjected to soil lead stress. The seedlings were cultivated outdoors in pots for five weeks. Results showed that total lead content, superoxide radical (O2*-) and lipid peroxidation productions in the leaves increased with the increase of extraneous lead in soils. Significant correlation was found between the O2*- and lipid peroxidation productions (r = 0.973, p < 0.01). Activities of superoxide dismutase (SOD), guaiacol peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX) were activated. Moreover, HSP70 and HSP60 increased more sensitively than the other parameters in response to lead stress. This indicates lead might cause lipid peroxidation in the leaves by mediation of O2*-. The activated antioxidative enzymes alleviated the stress and damage of reactive oxygen species. HSP70 and HSP60 are potential biomarkers for early warning of lead-contaminated soils. Additionally, concentrations of soil lead and related biological parameters should be combined. PMID- 19186837 TI - [Comparison of the treatment performance in fresh and mature landfill leachates by Fenton process]. AB - The experiments focused on comparing the removal efficiencies of organic matters in fresh and mature landfill leachate by Fenton treatment. The changes in dissolved organic matter before and after Fenton treatment were assessed using gross organic parameters such as organic compound relative molecular mass (Mr) distribution, chemical oxygen demand (COD), total organic carbon (TOC) and the ratio of COD to TOC. The results from this study demonstrate that high Mr fractions of mature landffill leachate are more easily converted into dissolved small Mr fractions (Mr < 10(3)) by Fenton reaction than those of fresh landfill leachate, since the average Mr of mature landfill leachate is higher than that of fresh landfill leachate and the Mr > 10(5) fraction of the mature leachate account for 64% of the total mass. Operating parameters such as pH, [Fe2+] and [H2O2]/[Fe2+] demonstrate that the COD removal efficiency of the fresh landfill leachate (from 61% to 84%) is higher than that of the mature landfill leachate (from 31% to 60%), while it is converse for TOC (from 85% and 52% to 91% and 80% for the mature and fresh landfill leachate, respectively). Oxidation seems to play a more active role in COD reduction than coagulation whatever for the mature or fresh landfill leachate. Oxidation efficiency apparently controls coagulation efficiency, so that high oxidation efficiency may cause relatively low coagulation. The mass balances indicate that TOC removal by oxidation (TOCoxid) is more than COD removal by oxidation (CODoxid) for the mature landfill leachate while TOCoxid is less significantly than CODoxid for the fresh landfill leachate. The COD/TOC ratios are 1.17 and 1.58 for the initial mature landfill leachate and fresh landfill leachate, respectively, but they are 1.96 and 0.68 after Fenton treatment by oxidation, respectively. This phenomenon suggests that complete oxidations occur in the mature landfill leachate while both partial and complete oxidations occur in the fresh landfill leachate. The changes of CODcoag and TOCcoag illustrate that the oxidation state of the organic matters removed by coagulation in the mature landfill leachate is significantly lower than in the fresh landfill leachate. PMID- 19186838 TI - [Contaminants natural attenuation of beitiantang landfills leachate polluted sites]. AB - Redox zones and the natural attenuation of pollutants in landfill leachate polluted site through analyzing data monitored around the Beitiantang Landfills of Beijing City. The results indicated that redox zones existed in Beitiantang Landfill leachate polluted sites, and 5redox zones were classified depended on distribution rules and characteristics of indicative substrates, namely methanogenic zone, sulfate reduction zone, iron reduction zone, nitrate reduction zone and oxygen reduction zone. Attenuation efficiencies of different organic contaminants in redox zones were different. Content of volatile hydroxybenzene and cyanide in sulfate reduction zone were relative high, and they were 0.005 and 0.019 microg/L, respectively. Heavy metal content, such as Cr, Pb, Ni, As, Cu, Cd, Zn and Mn were relative high in methanogenic zone, and they were 25.11, 33.82, 29.93, 3.18, 2.3, 0.1, 283.1, 1220 microg/L, respectively. Therefore, redox zones played an important role on the attenuation of pollutants. PMID- 19186839 TI - [Redox zones and its functional bacteria in a contaminated landfill site]. AB - A column filled with sandy soil was constructed to investigate biogeochemical process of leachate polluted zones. Experimental results demonstrated that four sequent redox zones appeared in pollution plume. The zones can be named sulfate reduction zone, iron reduction zone, nitrate reduction zone and oxygen reduction zone, ranges of them were 0-27 cm, 27-62 cm, 47-74 cm, 74-91 cm. In the redox zones bacterial community structure changed, and the preponderant bacteria were sulfate reduction bacteria (SRB), iron reduction bacteria (IRB), nitrate reduction bacteria (NRB) correspondingly, however there were other bacteria in the redox zones. Distribution of redox zones and functional bacteria means that there is not a significant boundary between redox zones, moreover one redox zone overlaps others. Evolvement of functional bacterial community brings the redox zones substitute. PMID- 19186840 TI - [Characteristics and sources of elements of atmospheric particles before and in heating period in Beijing]. AB - In order to study the characteristics and sources of atmospheric particles before and in heating period, samples of atmospheric particles were collected in November of 2006. Concentrations of elements in particles were determined with ICP-MS. The results shows that concentrations of As, Se, Mo, Cd in heating period are more than twice of those before heating period, and there is sharp increasing of concentrations of Zn, Pb, Tl, K, Se, As, Cu, Cd, Ag in fine particles in heating period. Furthermore, Zn, Na are associated with finer particles in heating period than those before heating period. Factor analysis on the chemical composition of particles shows that contribution of combustion and biomass burning goes up and contribution of crust goes down in heating period. PMID- 19186841 TI - [Catalytic purification of NO over active carbon fiber-loaded La2O3 catalysts]. AB - The active carbon fiber (ACF) loaded different contents of La2O3 catalysts were prepared, and their catalytic activities and durability for the purification of NO were investigated, and for comparison, those of ACF and HNO3/ACF for the title reaction were also investigated. The orthogonal experiment results showed that the best volume fraction of oxygen in the mixed gas was 5%, and the best air speed of mixed gas was 5000 m3 (m3 x h)(-1). The catalytic experiment results showed that the activity of HNO3/ACF was better than that of ACF when the temperature was below 414.08 degrees C, but the activity durability of HNO3/ACF was inferior than that of ACF. The activity and the activity durability of ACF could be markedly increased when it was loaded La2O3 catalyst, the activity center of the catalyst was La2O3, and the La2O3 catalyst best load amount was 20%, the activity of 20% La2O3/ACF would above 95% when the temperature was higher than 350 degrees C, and its activity durability was about 8 h, which was superior than most of the catalysts that had been reported. The results of FT-IR and TG experiments also showed that the NO purification principle of ACF and HNO3/ACF was different from that of La2O3/ACF, when NO was purified by La2O3/ACF, the competition reactions took place between CO and NO. PMID- 19186842 TI - [Development of the driving cycle-based mobile emission model (DCMEM)]. AB - Vehicle specific power (VSP) and engine stress (ES) were introduced to reflect the driving characteristics of vehicles in China. Emission rates of 60 BIN based on VSP and ES for light-duty vehicles were calculated on the basis of the on board emission test data in 5 cities of China. The driving cycle-based mobile emission model (DCMEM) was developed to simulate CO, NOx and HC emission factors of light-duty vehicles for different cities or driving cycles. The differences in CO and NOx emission factors between the DCMEM simulation results and the emissions testing results were less than 10%, and less than 18% for HC. PMID- 19186844 TI - [Z2 appliance: the pre-adjusted appliance based on Chinese normal occlusion]. AB - Z2 appliance is the pre-adjusted appliance designed for Chinese orthodontic patients. The prescription of the appliance is based on Chinese normal occlusion, which is much different from the West in the first and the second orders as well as the third one. The appliance routinely includes 20 brackets and 8 molar buccal tubers with 3 standard arch forms. Clinically, continued light force is used in whole treatment. The side-effects such as forward tipping of incisors, bite deepening and loss of molar anchorage are reduced further due to fewer tips built into the anterior brackets as well as lower friction elastometric modules used during aligning and leveling. In condition of arch are leveled completely, 0.48 mm x 0.64 mm stainless steel archwire with 1.47 N retraction force is the best combination for sliding mechanics, which is proved by 3D nonlinear finite element study. Self drilling micro-screw is used for maximum anchorage. In finishing stage 0.53 mm x 0.64 mm NT arch wire is added in order to get full torque expressing. The research of Chinese pre-adjusted appliance has been lasted for more than 10 years in the department and clinical studies on Z2 appliance show that with minimal wire bending, treatment is more efficient and result is high quality and more consistent for Chinese orthodontic patients. PMID- 19186843 TI - [Characteristic and prospects of heterotrophic nitrification]. AB - Nitrification is a key step in the global nitrogen cycle. Classically, nitrifying bacteria are chemoautotroph. Recently, bacteria that have the ability of heterotrophic nitrification have been intensively studied as potential microorganisms that may be used to overcomeproblems inherent in the conventional method. This review gives a broad overview of the current status of heterotrophic nitrification including the heterotrophic species which nitrify actively, optimal condition for nitrification, heterotrophic nitrification pathway, enzymatic system and isolation methods. Also, the promising prospects of heterotrophic nitrification are especially introduced. PMID- 19186845 TI - [Expression of mammalian target of rapamycin and p70 S6 kinase in rabbit periodontal tissues remodeling during orthodontic tooth movement]. AB - OBJECTIVE: To study the role of mammalian target of rapamycin (mTOR) and p70 S6 kinase (p70 S6K) in periodontal tissues remodeling during orthodontic tooth movement. METHODS: Twenty-four rabbits were chosen to establish rabbit models for the study. The right maxillary teeth of each animal treated by orthodontics were the test sides, and the untreated left teeth were the control sides. The animals were sacrificed at 3, 5, 7, 14 d, respectively. The prepared tissue specimens were processed for the study of the change in the histologic morphology with HE staining and the expression of p70 S6K in periodontal tissues by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot techniques. RESULTS: In the test side, the tranecula of bone became porous and cells arrayed disorderly, resorption of alveolar bone was observed in histological study. There were many holes in the alveolar bone, osteoblasts were occasionally observed. RT PCR showed that the expression of p70 S6K mRNA dramatically changed at 3 d. The expression of p70 S6K mRNA in treating periodontal tissues was higher than the control side, especially at 7 d, and then decreased. Compared with the control side, there was significant difference in statistical analysis (P<0.01). The expression of Western blot was same as the RT-PCR. CONCLUSION: Higher expression of mTOR and p70 S6K during orthodontic tooth movement especially plays an important role in the process of periodontium remodeling. PMID- 19186846 TI - [Finite element analysis of the effects of implant thread locations on stress distribution]. AB - OBJECTIVE: To investigate the effects of different implant thread locations on stress distribution in implant and bone under concentrated load, and discuss the best design of location of implant thread. METHODS: The stress in implant-bone interface of implants with different thread locations (model A, B, C, D), which were loaded in axial and oblique 450 to axle, were analyzed by Solidworks 2005 plus and Cosmos/works 7.0 software. RESULTS: The stresses in cervical cortical bone of implant with model C were lower than others. But under oblique load, the stresses of implant and cancellous bone in model C were higher than model A. The stresses distribution of model B was obviously concentrated, and under axial load, all kinds of stresses in this model were higher than others. The stress distribution in model A and D were more even, the stress concentrated more on implant neck, upper part of cortical bone and the lowest thread on implant. The stresses in interface under oblique load were much higher than those under axial load. CONCLUSION: The location of thread has effect on stress distribution in implant-bone interface. More attention should be paid to it when we design implants, and we should try to avoid oblique load in implant prosthodontics. PMID- 19186847 TI - [Effects of mechanical stress on the formation and expression of Cbfalpha1 in vitro]. AB - OBJECTIVE: To evaluate the effects of mechanical stress on the formation and expression of core binding factor alpha1 (Cbfalpha1) in MG-63 cells cultured on titanium in vitro. METHODS: MG-63 cells cultured on the titanium were subjected to a centrifugal force (2.205 N) 15 min per 4 hours and collected after 4, 8 and 12 hours. The formation and expression of Cbfalpha1 were examined by immunofluorescence staining and reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Both the cells with or without centrifugal force created the fluorescence in the nucleus and the immunofluorescence intensity of Cbfalpha1 in MG-63 cells with centrifugal force were higher than those without centrifugal force (P<0.05). Meanwhile, both the cells with or without centrifugal force expressed the mRNA of Cbfalpha1 and the relative mRNA level of Cbfalpha1 in MG-63 cells with centrifugal force were higher than those without centrifugal force, and the differences were great significant (P<0.05). CONCLUSION: Mechanical stress are beneficial to the formation and expression of Cbfalpha1. PMID- 19186848 TI - [Expression and significance of vascular cell adhesion molecule-1 in human oral squamous cell carcinoma]. AB - OBJECTIVE: To study the expression and the location of vascular cell adhesion molecule-1 (VCAM-1) gene and its clinical significance in human oral squamous cell carcinoma (OSCC). METHODS: In situ hybridization, PV-9000 polymer detection system for immunohistochemical staining was used to detect the expression and the location of VCAM-1 mRNA and VCAM-1 protein in 48 cases of OSCC and 10 cases of normal controls. Statistical analysis was performed using chi-square test in SPSS 13.0. RESULTS: VCAM-1 protein was mainly expressed in tumor cell cytoplasm and membrane, VCAM-1 mRNA was mainly expressed in tumor cell cytoplasm. The expression rate of VCAM-1 mRNA and VCAM-1 protein was significantly higher in OSCC than that in normal oral mucosa (P<0.01). The expression of VCAM-1 mRNA was positively correlated with that of VCAM-1 protein (P<0.01). In the clinicopathologic factors, lymph node metastasis and depth of infiltration were closely correlated with VCAM-1 expression (P<0.01). The expression of VCAM-1 was significantly higher in tumor with lymph node metastasis than in tumor without lymph node metastasis (P<0.01). CONCLUSION: Overexpression of VCAM-1 gene in OSCC may play a potential role in the development of OSCC. The overexpression of VCAM 1 gene in OSCC may be related to the tumor infiltration and metastasis. PMID- 19186849 TI - [Experimental study of Ti-24Nb-4Zr-7.9Sn alloy after treated with micro-arc oxidation in vivo]. AB - OBJECTIVE: To study the biocompatibility of Ti-24Nb-4Zr-7.9Sn (TNZS) ahoy treated with micro-arc oxidation (MAO). METHODS: The tibia bones of New Zealand rabbits were used to build the animal model. TNZS and MAO-TNZS samples were implanted into one side of tibia, pure titanium samples were implanted into the other side as control. After 4 and 26 weeks, radiographs and HE staining technique was used to observe the dynamic remodeling process of bone-implant interface. RESULTS: As the cure time increased, it was showed well biocompatibility of all implants. X ray indicated that there was no permeable area produced around the three different materials at each time point. The density of bone matrix and arrangement of bone trabecula was almost the same as in the host bone. It was revealed by histological examination that the MAO-TNZS greatly prompted the bonding ability between implant and surrounding hard tissues. Four weeks after implantation, fine attachment was found at the bone-implant interface of all the implants and the fibrous tissue at the interface was gradually remodeled to form new bone. Twenty-six weeks later, MAO-TNZS showed that a biological fixation was created between bone and oxidation layer, while a layer of fibers formed between non-coated TNZS and titanium implants surrounding bone. CONCLUSION: The Ti-24Nb 4Zr-7.9Sn after treated with micro-arc oxidation shows good biocompatibility and can stimulate the bone growth in the bone-implant region, which provides support for clinical usage tests of TNZS alloy as implant after treated with micro-arc oxidation. PMID- 19186850 TI - [Differentiation and functional expression of highly purified osteoclast-like cells in vitro]. AB - OBJECTIVE: To establish a culture method for a large amount of highly purified osteoclast-like cells in vitro. To investigate the gene expression of some osteoclast marker enzymes. To lay the foundation for the further study of the signal path on the differentiation and formation of osteoclast-like cells. METHODS: The bone marrow mononuclear cells of rat were treated with 30 ng/mL macrophagecolony-stimulating factor (M-CSF) and 50 ng/mL receptor activator of NF kappaB ligand (RANKL) and cultured for 6 days. After culturing, cells were evaluated by morphology observation, tartrate-resistant acid phosphatase (TRAP) staining, Giemsa staining, pit staining, and the gene expression of some osteoclast marker enzymes. RESULTS: The TRAP-positive mononuclear cells were more frequently observed than the multinucleated cells and pit staining could be seen on the dentine slice. The transcription expression of TRAP, matrix metalloproteinase-9 (MMP-9), membrane-type1-matrix metalloproteinase (MT1-MMP) and cathepsin K were detected by RT-PCR. CONCLUSION: The cooperation of M-CSF and RANKL could induce a large amount of highly purified osteoclast-like cells formation in rat bone marrow culture. The typical characteristics of osteoclast like cells were demonstrated and the enriched osteoclast-like cells expressed TRAP, MMP-9, MT1-MMP and cathepsin K. PMID- 19186851 TI - [The expression of Maspin protein in oral squamous cell carcinoma and its significance]. AB - OBJECTIVE: To investigate the role of Maspin protein in the development of oral squamous cell carcinoma (OSCC), with the effort to realize practical diagnosis use of them. METHODS: The expression status of Maspin protein in 45 cases of OSCC, 33 cases of paratumor tissue and 15 cases of normal tissue was examined by immunohistochemical staining. RESULTS: The positive rates of Maspin protein were 86.67% (13/15), 72.73% (24/33) and 37.78% (17/45) in normal tissue, paratumor tissue and OSCC. The expression of Maspin protein increased respectively in OSCC, paratumor tissue and normal tissue. The expression were different between OSCC and normal tissue, OSCC and paratumor tissue, paratumor tissue and normal tissue (P<0.05). The expression of Maspin protein was correlated with lymph node metastasis, histological grade (P<0.05), but not with TNM staging (P>0.05). CONCLUSION: Maspin protein may play an important role in the development of OSCC. The expression level of Maspin protein detected in early time might be a useful prognostic marker for the diagnosis of lymph node metastasis. PMID- 19186852 TI - [Detection and prevention of early caries after fluoride dentifrice application using quantitative light-induced fluorescence in vivo]. AB - OBJECTIVE: To evaluate the capability of quantitative light-induced fluorescence (QLF) for monitoring the remineralization of lesions lengthways and distinguishing the effect of different fluorides. METHODS: Following baseline early caries examination, 305 school students (age from 11 to 14 years old)were qualified from Chengdu area. The schools in which the subjects studied were randomized into three groups: NaF group, MFP group and nonfluoride group. The subjects of three groups brushed the teeth with 1450 mg/L sodium fluoride dentifrice, 1450 mg/L sodium monofluorophosphate dentifrice and non-fluoride dentifrice, respectively. QLF images of early lesions on smooth surfaces of the maxillary anterior teeth were taken at baseline, three and six months after the initiation of experiment. These images were analyzed by the trained examiner with the area of lesion (Area), fluorescence loss (deltaF) and gross fluorescence loss quantity (deltaQ). RESULTS: 296 school students completed the study. After three months, the lesions on smooth surfaces got better in all three groups. The change of Area, deltaF and lg deltaQ amongst three groups had no statistical significance (P>0.05). After six months, the lesions on smooth surfaces also got better in all three groups than at baseline. The change of Area, deltaF and lg deltaQ of the lesions in NaF group and MFP group exhibited significant decreases than that of no-fluoride dentifrice group (P<0.05). But no significantly difference was found between NaF group and MFP group (P>0.05). CONCLUSION: As a diagnostic method which could quantify the miner content, QLF system has the capability of monitoring the variations of lesions lengthways. PMID- 19186853 TI - [Application of multislice helical computed tomography in maxillofacial distraction osteogenesis]. AB - OBJECTIVE: To develop the method and investigate the value of the application of multislice helical computed tomography (CT) in the maxillofacial distraction osteogenesis. METHODS: Twelve cases of maxillofacial distraction osteogenesis (3 in zygoma, 2 in maxilla, and 7 in mandible) were scanned by the multislice helical CT, then the reconstruction of three dimensional and fault image and the relative measurement were carried out to study the effect of distraction and osteogenesis. RESULTS: The three dimensional image provided clear and high resolution stereoscopic view that the effect of distraction osteogenesis could be evaluated easily and exactly. The fault image could be set up in private computer by Mimics and the measurements, such as distance, thickness and density, could be performed as expected. The effect of 12 cases was satisfied, and the average distracted distance was 16.73 mm. CONCLUSION: Basing on multislice helical CT scanning, the high resolution image reconstruction and relative measurement data could be obtained which could meet the need of clear and direct observation. Its application in the maxillofacial distraction osteogenesis is expecting and promising. PMID- 19186854 TI - [Study on mandibular rotation in 13 female subjects aged from 9.5 to 15.5]. AB - OBJECTIVE: To evaluate the mandibular rotation in females aged from 9.5 to 15.5 years. METHODS: The sample was obtained from Mathews's implant study with longitudinal cephalometric records of 13 untreated Caucasian female subjects. The cephalograms taken annually from 9.5 to 15.5 years old were traced and analyzed with Dentofacial Software. Paired-t test was performed to identify the significant growth changes in different age groups. RESULTS: Significant change in lower anterior facial height, posterior facial height, ramus length and mandibular implant plane angle were noted in aged 9.5-11.5. Significant change in lower anterior facial height, posterior facial height and ramus length were noted in aged 11.5-13.5. Significant change in mandibular plane angle and mandibular implant plane angle and significant increase in posterior facial height were observed in aged 13.5-15.5. CONCLUSION: The results of this study verify that remodeling on the lower border of mandible influences total mandibular rotation. PMID- 19186855 TI - [Clinical research of craniofacial bone autograft using mandibular outer cortex]. AB - OBJECTIVE: To illustrate the volume changes of mandibular outer cortex after been grafted to different recipient sites of facies cranii. METHODS: Six cases underwent onlay bone graft to the angle and body part of mandible and malar surface simultaneously by using mandibular outer cortex. Three dimensional computed tomography (3D-CT) datum of immediate postoperative and 6 months postoperative of each case were collected systematically. By true-up and dissection techniques based on 3D-CT, volume changes of bone graft at different recipient sites were observed and analyzed 6 months postoperative. RESULTS: 6 months after onlay bone grafted, bone resorption occurred. To different recipient sites, bone resorption rate was unequal. At the mandible area, bone resorption rate was 20.8% +/- 7.2%, the main site of resorption was at the lower and posterior border of mandible. At the anterior part of maxilla, bone resorption rate was 11.2% +/- 2.3%. Statistics showed significant difference of resorption rate between the two sites (P<0.05). CONCLUSION: Bone resorption of mandibular outer cortex after onlay graft treatment is variant according to different craniofacial recipient sites. The difference of mechanical environment at variant recipient sites is considered to be an influencing factor. Quantization of bone resorption rate can guide a better clinical use. PMID- 19186856 TI - [A primary study of color tolerance of anterior tooth crown prosthesis]. AB - OBJECTIVE: To study the color tolerance of anterior ceramic crown and seek an effective approach for color-matching of oral prostheses. METHODS: 30 single maxillary incisor ceramic prostheses and the corresponding nature teeth were measured by a digital camera in a steady environment for image taking and color measurement, which based on the standard recommended by CIE. The color tolerance was analyzed in two groups based on satisfaction and dissatisfaction of prostheses color-matching. RESULTS: The deltaE of satisfaction group was 2.5503, while the deltaE of dissatisfaction group was 4.0772. There was significant difference between chromatic aberration of satisfaction group and dissatisfaction group (P<0.01). When the lightness and saturation of prosthesis were higher than nature teeth, there was no significant difference between deltaL*, deltab*, deltaC* of satisfaction group and dissatisfaction group (P>0.05). When the lightness and saturation of prosthesis were smaller than nature teeth, there was significant difference between deltaL*, deltab*, deltaC* of satisfaction group and dissatisfaction group (P<0.01). There was significant difference between deltaa*, deltaH of satisfaction group and dissatisfaction group (P<0.05, P<0.01). CONCLUSION: Control of color difference is the key for shade matching of anterior tooth prosthesis. Applying the color tolerance is beneficial to improving the prosthesis quality. PMID- 19186857 TI - [Clinical evaluation on the root canal therapy bypassing intracanal separated instruments]. AB - OBJECTIVE: To study and evaluate the clinical effect of root canal therapy bypassing intracanal separated instruments. METHODS: 32 teeth were selected as the test group, from which the intracanal separated instruments couldn't be taken out, adopted the method of root canal preparation and obturation bypassing broken instruments. Meanwhile, 13 teeth were selected as the control group, in which the fractured instruments couldn't be taken out from root canal, and not be cured bypassing broken instruments. 30 teeth were selected as the conventional therapy group, in which the instruments were not fractured, the routine root canal therapy were adopted. The clinical efficacy of three groups were observed and evaluated through periapical index and clinical examination two years later. RESULTS: After two years, 25 teeth were followed up in test group, the success rate of root canal therapy was 80.00%; 11 teeth were followed up in control group, the success rate of root canal therapy was 36.36%. All of the failed cases in two groups were the teeth with broken instruments in tip of root canal. In conventional therapy group, 22 teeth were followed up, and the success rate of root canal therapy was 90.91%. CONCLUSION: Root canal therapy bypassing intracanal separated instruments is a feasible and effective method in treating the teeth with broken instruments. PMID- 19186858 TI - [A comparison study on the effect of self-etching adhesive and phosphoric acid fissure sealant in children]. AB - OBJECTIVE: To compare the retention and caries prevention of fissure sealant on the primary molars etched with self-etching adhesive and phosphoric acid in children. METHODS: 43 children younger than 42 months with more than 4 decayed teeth were studied. The split-mouth design was used in this study. The second and/or first primary molars were randomly assigned into self-etching adhesive group in one side of mouth, and phosphoric acid group in the other side. Sealants were applied on 69 pair primary molars according to manufacturer' s instruction with dental cotton-roll isolation. The children were followed up every 3 months, and the sealants were observed and evaluated. In the case caries occurred, the teeth were restored. RESULTS: After 3 months, the retention rate was 93% in self etching adhesive group and 100% in phosphoric acid group. At 6, 12 and 18 months, the retention rate of self-etching adhesive group was statistically lower than that of phosphoric acid group (P<0.05) by Wilcoxon signed ranks test. Caries occurred in both groups and self-etching adhesive group had more caries than phosphoric acid group. Though its retention rate was low, self-etching adhesive was suitable for young children because it was timesaving and its bonding procedures were simple without discomfort rinsing. CONCLUSION: The retention rate of sealant with self-etching adhesive was lower than that with phosphoric acid. The phosphoric acid is suitable for pit and fissure sealant. If self-etching adhesive is used for sealing primary molars of young children for a limited period of time, it should be reviewed every 3 months. PMID- 19186859 TI - [Fracture resistance and failure modes of endodontically treated human teeth restored with four different post-core systems]. AB - OBJECTIVE: To compare the fracture resistance and failure modes of endodontically treated human upper premolars restored with different post-core systems. METHODS: 32 extracted human upper premolars were endodontically treated and crowns were sectioned at 2 mm above the labial cement enamel junction (CEJ). The teeth were randomly and equally divided into 4 groups: Group A, teeth restored with cast metal post and metal crown; group B, Tenax Fiber White fiber post and metal crown; group C, EverStick fiber post 1.5 mm in diameter and metal crown; group D, EverStick fiber post 1.5 mm in diameter and add another 1.2 mm diameter EverStick fiber post and metal crown. All the teeth were embedded in acrylic resin blocks, and were subjected to a compressive load at 1 mm/min crosshead speed which delivered at a 45 degrees to the long axis until the first sign of failure was noted. The fracture load and the mode of fracture were recorded. RESULTS: Fracture resistances of the four groups of restored teeth were not significantly different (P>0.05). However, fracture modes in fiber post groups were nearer to CEJ than cast metal post group. CONCLUSION: Fracture resistance of endodontically treated teeth restored with EverStick fiber post is enhanced that it can meet the clinical need, although the flexural resistance of EverStick fiber post itself is the weakest. Fracture modes of all fiber post groups are more favorable than cast metal post group. PMID- 19186860 TI - [The application of cone-beam computed tomography and nine partition method on alveolar bone for locating the unerupted teeth]. AB - OBJECTIVE: To investigate the clinical value of cone-beam computed tomography (CBCT) and nine partition method on alveolar bone for locating the relative orientation of the unerupted teeth. METHODS: 125 bone unerupted teeth of 99 cases were collected. The bone unerupted teeth zones were scaned by CBCT. The dental corona and root apex of these bone unerupted teeth were classified by the new nine partition method. RESULTS: In 125 bone unerupted teeth, 107 teeth located in maxilla, 18 teeth located in mandible. More frequent appearances (35.6%) in the alveolar bone were inner-middle and outer-superior. The representation of 125 bone unerupted teeth seen in surgery was the same with that of the CBCT before exodontia. The accurate rate of diagnosis was 100%. CONCLUSION: CBCT is the most accurate and effective methods to determine the authentic orientation of the bone unerupted teeth. The nine partition method of alveolar bone has clinical value of accurate authentic orientation of the bone unerupted teeth in oral and maxillofacial surgery and orthodontics. PMID- 19186861 TI - [The characteristics of masticatory muscle activity in bruxers]. AB - OBJECTIVE: To evaluate the effects of bruxism on masticatory muscle electromyographic (EMG) activity. METHODS: Twenty-four bruxers and sixteen asymptomatic control subjects were included through questionnaire and clinical examination. EMG activity was recorded by placing surface electrodes on bilateral anterior temporalis (TA), masseters (MM), anterior digastrics (DA) and sternocleidomastoid (SCM) muscles. EMG activities at rest, during maximal voluntary clenching in intercuspal position and swallowing were recorded by means of Bio PAK system. RESULTS: EMG activities of TA and MM at rest were significantly higher in bruxism group than in control group (P<0.05). When subjects clenched their teeth in intercuspal position, the activities of TA and MM were much lower in bruxism group than in control one (P<0.05). EMG activity during swallowing was no significant difference between the two groups. The asymmetry index of bilateral TA and MM in bruxism group was a little higher than the control group, but there was no significant difference between the two groups (P>0.05). CONCLUSION: Masticatory muscle dysfunction of bruxers is mainly represented as higher potential in postural position and lower potential during maximal voluntary clenching in intercuspal position of anterior temporalis and masseters. PMID- 19186862 TI - [Psychological impact on the adolescent patients at the beginning of the fixed orthodontic treatment]. AB - OBJECTIVE: To assess whether there is a short negative psychological influence on adolescent patients at the beginning of the fixed orthodontic treatment. METHODS: 150 patients (average 14.8 years old) were selected. All the patients accepted the fixed appliance treatment. They completed a questionnaire regarding anxiety and depression at the first day when they came to the hospital (T1) and 7 days after fixed appliance insertion (T2). 129 effective questionnaires were received. The scales of anxiety and depression of subjects were assessed according to the questionnaires. RESULTS: Comparing the scales of questionnaires before treatment (T1) and 7 days after placement of fixed appliance (T2), there was a significant increase in anxiety and depression scales in female patients, extraction cases and patients who were unwilling to see an orthodontist. CONCLUSION: There is a certain extent of negative psychological influence on adolescent patients during fixed orthodontic treatment. At the first week after the placement of fixed appliance, three kinds of subjects, female patients, extraction cases and patients who were unwilling to see an orthodontist would suffer from anxiety and depression in emotional reflection. PMID- 19186863 TI - [Relationship of porphyromonas gingivalis PG0717 gene with chronic periodontitis]. AB - OBJECTIVE: The aim of the present study was to investigate PG0717 gene of Porphyromonas gingivalis (P. gingivalis) in subgingival plaque of the chronic periodontitis patients and periodontal healthy subjects, and to find out the relationship of detection rate of PG0717 and periodontal clinical parameters. METHODS: A total of 540 subgingival plaque samples were collected from 180 subjects including chronic periodontitis (CP) patients (n=90) and periodontal healthy individuals (n=90). The periodontal clinical parameters including probing depth (PD), clinical attachment loss (CAL), and bleeding on probing (BOP) were estimated by Florida probe. The extracted DNA samples of P. gingivalis positive was amplified with the sequence specific primers designed to obtain the PG0717 gene. RESULTS: In subgingival plaque of P.gingivalis positive, the detection rate of PG0717 in CP group was significantly higher than that in periodontal healthy subjects (56.22% versus 41.27%, chi2=4.50, P<0.05). The detection rate of PG0717 in CP group showed the increasing tendency in accordance with the depth of PD and CAL. A higher detection rate of PG0717 was observed in the sites of BOP positive than that in BOP negative (57.73% versus 14.29%, chi2=42.01, P<0.01). CONCLUSION: The findings suggest that the PG0717 gene may influence the virulence of P. gingivalis. PMID- 19186864 TI - [Interleukin-8 regulations of oral epithelial cells by porphyromonas gingivalis with different fimA genotypes]. AB - OBJECTIVE: The expression of heterogenic virulence properties depends on its clonal diversity. The aim of the study was to investigate the mechanism of interleukin-8 (IL-8) regulations of oral epithelial cells by challenge of Porphyromonas gingivalis (P. gingivalis) with different fimA genotypes, discuss the relation between fimA genotype and its pathogenicity. METHODS: P. gingivalis ATCC 33277 (type I), W83 (type IV), 47A-1 (type IV) were assessed for their inductions of IL-8 expression in human oral epithelial cells (KB cell line, ATCC CCL-17). KB cells without stimulation of P. gingivalis were used as control group. IL-8 mRNA expression was de termined by reverse transcription polymerase chain reaction (RT-PCR) at different time intervals (1, 3, 6, 24 h) following continuous co culture of bacteria with KB cell line, and IL-8 protein levels in culture supernatant was determined by enzyme-linked immunosorbent assay. RESULTS: IL-8 mRNA levels were up-regulated and reached its high peak at 1 h following both genotypes infection, then decreased to base level till 24 h. Attenuation of IL-8 protein levels was down-regulated when KB cell co-cultured with both genotypes for 3 h till 24 h, and type IV was lower than type I. IL-8 and IL-6 mRNA expression were not consistent with their protein levels, which indicated post-transcriptional regulations. CONCLUSION: fimA genotypes of P. gingivalis are related with the effect of IL-8 inductions, which indicates fimA genotype is associated with pathogenesis of P. gingivalis. PMID- 19186865 TI - [Effects of mineralization liquid on rat's osteoblast proliferation and differentiation]. AB - OBJECTIVE: To compare the rat osteoblast proliferation and differentiation in the mineralization condition and non-mineralization condition, and to prove the feasible time of adding the mineralization liquid to the cell surroundings. METHODS: SD rat's calvarial osteoblast was primary cultured and passaged to the 4th generation. After proliferation stabilized, the 4th generation rat's osteoblast was cultured in the mineralization condition and non-mineralization condition. The cellular modality was observed by inverted microscopy analyzing system. Proliferation was described by MTT chromatometry and growth curve. Expression of alkaline phosphatase (ALP) was proved by alkaline phosphatase staining, mineralized nodus alizarin red staining and enzyme-linked immunosorbent assay (ELISA). The cell apoptosis and proliferation were examined by flow cytometry. RESULTS: The proliferation of osteoblast went to the flat stage about in the 8th day. After in the mineralization condition 3 weeks, a number of mineralized nodus formed. After the osteoblast proliferation almost stabilized, the mineralization group and the non-mineralization group had the similar proliferation index (PI), but in the mineralization group, the expression of alkaline phosphatase of the osteoblast was more and kept longer than the last group. CONCLUSION: After the osteoblast proliferation almost stabilized, adding the mineralization liquid would not influence the proliferation, but accelerate the osteoblast alkaline phosphatase expression and mineralized nodus amount. It proved that the feasible time to add the mineralization liquid was after the cell proliferation almost stabilized. PMID- 19186866 TI - [Experimental study of preparing curved root canals with different instrument]. AB - OBJECTIVE: To compare the shaping ability and the influence on apical foramen among hand ProTaper, stainless steel K-files and rotary ProTaper in preparing different curved root canal. METHODS: Forty simulated resin root canal blocks were randomly divided into four groups and prepared by hand ProTaper, stainless steel K-file and rotary ProTaper, respectively. Of them, 12 blocks in group A, B, C consist of six 200 curved root canals and six 30 degrees curved root canals each group. The curvature of the other 4 blocks in group D was less than 5 degrees. Taking photos of the models to the root canal orthotopically and apical foramen using digital camera before and after instrumentation. Finally, the transportation of root canal and the size of apical foramen were analyzed using special image software Auto-CAD. RESULTS: The transportation of center in group B was the highest than that in group A and group C (P<0.05). In some portions of root canal, the transportation of center in group C was higher than that in group A. The size of the apical foramen in group B was significantly bigger than the other groups and the size of the apical foramen in 30 degrees root canal was significantly bigger than that in 20 degrees root canal after instrumentation (P<0.05). There was no significantly different between group A and group C, though the size of apical foramen in group C was bigger than that in group A at the same curvature, and that in 30 degrees root canal was bigger than in 20 degrees root canal (P>0.05). CONCLUSION: Both of the two instruments engender root canal transportation, and curvature is the main reason of transportation. Comparing with stainless steel K-files, NiTi files can maintain the shape of the root canal and apical foramen well. PMID- 19186867 TI - [Effect of loop configuration on appliance stiffness of stainless steel round wire]. AB - OBJECTIVE: To investigate the effect of loop configuration on appliance stiffness of stainless steel round wire. METHODS: Some stainless steel round wires with various cross-sectional dimension (0.36, 0.41, 0.46 mm) were selected. Box loops, double horizontal loops, and double vertical loops were manufactured according to established parameters, and loading experiments were carried out with a micro orthodontic archwire testing apparatus. RESULTS: When the cross-sectional dimension and the span were identical, the appliance stiffness of the box loops, double horizontal loops, and double vertical loops increased successively. CONCLUSION: Configuration of loops exert a great influence on appliance stiffness, and the mechanical properties of box loop are optimal. The force generated by box loop was lighter, slowly decreasing, and easier to be controlled. The activation range of box loop (0.41 mm) was appropriate when the produced optimal moment was between 0.01-0.02 N.m. Box loop (0.41 mm) is preferred to rectify the individual tooth with mild or moderate mesiodistal displacement. PMID- 19186868 TI - [Differential analyses of mRNA expression of gtfs from Streptococcus mutans in different pH condition]. AB - OBJECTIVE: To determine the expression level of each gtf under different pH cultural conditions and to find the relationship between gtf expression levels with environmental pH in different strains of Streptococcus mutans (S.mutans). METHODS: S. mutans form clinical isolation with different extracellular polysaccharides (EPS) producibility and UA159 were selected. Their ability to produce EPS under pH5.5 and pH7 were tested. Then in two strains, the relative quantity of gtfA, gtfB, gtfC, gtfD's mRNA which were related to S. mutan's ability to produce EPC, were examined by real-time reverse transcription polymerase chain reaction (real-time RT-PCR) methods under different pH culture condition. RESULTS: At pH5.5, expression levels of gtfA, gtfB, gtfD were increased while that of gtfC were decreased in both strains, and that of gtfB, gtfC were higher in strain which produces more ECP. CONCLUSION: The expression levels of gtfs related closely to the cariogenicity of S. mutan. PMID- 19186869 TI - [Biologic analysis of recombinant human adenovirus-p53 injection in patients with oral leukoplakia ]. AB - OBJECTIVE: Advances in tumor biology and clinical trials indicate that p53 transfer is an alternative therapy for head and neck squamous cell carcinoma. The aim of this study is to evaluate the biologic activity of multiple intraepithelial injections of Ad-p53 in patients with dysplastic oral leukoplakia, which is the most common premalignant lesion of the oral squamous cell carcinoma. METHODS: From 2006 to 2007, 18 Chinese patients clinically and histopathologically diagnosed as dysplastic oral leukoplakia were recruited for this study. On a 15-day cycle, intraepithelial injections of Ad-p53 were administered once every three days at dose levels dependent upon lesion size/dose escalation sequence (1 x 10(8) vp). 24-48 h after the last injection, incisional biopsy were performed, and immunohistochemistry was used to examine the protein expression of P53 and P21(CIP/WAF). RESULTS: In the postreatment patients, P53 protein and P21(CIP/WAF) protein expression were significantly enhanced (100%, 89.9%, respectively) and statistical analysis revealed the expression of P53 protein had a positive correlation with that of P21(CIP/WAF) protein (r=0.598, P<0.01). CONCLUSION: Intraepithelial injections of gendicine is biologically active in patients with dysplastic oral leukoplakia. It may be a promising treatment for oral leukoplakia. PMID- 19186870 TI - [Establishment and identification of transiently expression system of bone marrow stromal cells modified by osteoprotegerin gene]. AB - OBJECTIVE: In oder to treat periodontitis by using tissue engineering and gene engineering technology, the article established an transient expression system of bone marrow stromal cells (BMSC) modified by osteoprotegerin (OPG) gene and detected its expression using eukaryotic secreted expression pSecTag2/B-OPG plasmid. METHODS: By solation and culture of BMSC in vitro, the identified recombined plasmid was transiently transfected into BMSC by Lipofectamine 2000 and OPG expression in BMSC was determined by RT-PCR and Western blot in 6 weeks. RESULTS: The fragments of the recombinant plasmid digested with Hind III, EcoR I and BamH I and examined by 10 g/L agarose electrophoresis, were consistent with predicted size. The sequence of OPG was identical to the sequence provided by GeneBank [gi:33878056]. OPG transcribing in BMSC was confirmed by RT-PCR and OPG sustainable expressing in BMSC was detected by Western blot in 39 days. CONCLUSION: The transiently expression system of BMSC modified by OPG gene was successfully established. PMID- 19186871 TI - [A case of repairing face slant diastema by rotating push of nasal bottom skin flap]. AB - We use Z type skin flap to repair face slant diastema by combining nasal bottom and infraorbital border. The therapeutic efficacy was satisfactory. The dislocated nostril, eversioned eyelid and lowered eyeball was resettled. PMID- 19186872 TI - [A case report of fixed restoration of hereditary opalescent dentin]. AB - Hereditary dentinogenesis imperfecta is a rare autosomal dominant hereditary disease. This article reported a case of hereditary opalescent dentin and discussed the clinical features and treatment. PMID- 19186873 TI - [A case of vertebral traumatic pseudoaneurysm]. AB - Pseudoaneurysm often occurred after trauma and infection. It was seen commonly on limb arteries but rarely on vertebral artery. The case was diagnosed two months after trauma. The pulsatile mass and vascular murmur found through preoperative examination confirmed from the vertebral artery. No complication was observed after surgery. PMID- 19186874 TI - [The human factor]. PMID- 19186875 TI - [The current hospital reform and the 21,000 position program for nursing in the hospital: it must be managed quickly]. PMID- 19186876 TI - [Motivations for personal and patient oriented health education for general nursing practice: health and body perception]. PMID- 19186877 TI - [Challenging behavior of patients with dementia: experiences and strategies of nurses]. PMID- 19186878 TI - [Procedures in a fire incident in a social facility: should the facility always be evacuated immediately?]. PMID- 19186879 TI - [Spanish influenza 1918/19 in Nurnberg: H1N1 and the consequences]. PMID- 19186880 TI - [Experiences with pastoral services in geriatric nursing: relationship nursing: more current than ever]. PMID- 19186881 TI - [Psychological observations on assisted death: motifs of health experts: patient will must be in first place]. PMID- 19186882 TI - [Aging in geriatric nursing: confronting personnel management challenges early]. PMID- 19186883 TI - [Forensic nursing in Germany? Nurses' perceptions of domestic violence]. AB - More than one of three women has been a victim of domestic violence at least once. Victims would like to have a well-informed contact person within the healthcare system who knows about support programs. In many countries that is the responsibility of the healthcare system and is called Forensic Nursing. Therefore, it is interesting to know how nurses in Germany perceive domestic violence and under what circumstances they could imagine taking on tasks in the fields of documentation and nursing. The data for this qualitative study was collected via four focus groups consisting of 38 nurses--3 men and 35 women--with work experience in a hospital. Nurses seem to have difficulties in recognising domestic violence. Whether the subject of domestic violence is addressed explicitly depends on the relationship built up between the patient and the nurses. Nurses do not necessarily take further steps. They could imagine providing help by listening actively, providing information about support programs and providing consulting services. Only occasionally nurses agree to document the case to be used as forensic evidence. Another open issue is appropriate remuneration. It is necessary to integrate that subject systematically into basic and advanced training on different levels of qualification. PMID- 19186884 TI - [Long-term preservation of competence: entitlement for vacation time]. PMID- 19186885 TI - [Prevention in nursing education--3: Prevention of aspiration: choking can happen quickly]. PMID- 19186886 TI - [From case to case: safety risk of medical products]. PMID- 19186887 TI - [Recognizing and understanding illness: pallor as the leading symptom]. PMID- 19186888 TI - [Implementing digital nursing care planning and documentation: increasing transparency and quality of the nursing process]. PMID- 19186889 TI - Effluent from bulk drug production is toxic to aquatic vertebrates. AB - Very high levels of a range of pharmaceuticals have been reported recently in the effluent from a wastewater treatment plant near Hyderabad, India. The plant serves approximately 90 manufacturers of bulk drugs that primarily are exported to the world market. Fluoroquinolone antibiotics were found at levels that are highly toxic to various microorganisms. Even though milligram-per-liter levels of drugs targeting human proteins also have been found, it is difficult to conclude whether these levels are sufficiently high to adversely affect fish or amphibians due to the lack of relevant chronic toxicity data for most human pharmaceuticals. To assess potential effects on aquatic vertebrates, tadpoles of Xenopus tropicalis were exposed to three dilutions of effluent (0.2, 0.6, and 2%) over 14 d, starting at developmental stage 51. Additionally, newly fertilized zebrafish (Danio rerio) were exposed to diluted effluent in 96-well plates for up to 144 h postfertilization (hpf). The tadpoles' body lengths, weights, and developmental stages were recorded, whereas a larger number of sublethal and lethal endpoints were studied in the zebrafish. A 40% reduced growth of the exposed tadpoles was demonstrated at the lowest tested effluent concentration (0.2%), indicating potent constituents in the effluent that can adversely affect aquatic vertebrates. The median lethal concentration (LC50) for zebrafish at 144 hpf was between 2.7 and 8.1% in different experiments. Reduced spontaneous movements, pigmentation, and heart rate were recorded within 48 hpf at 8 and 16% effluent concentrations. Treated effluent from a plant that serves as an important link in the global supply chain for bulk drugs is thus shown to cause adverse effects to aquatic vertebrates even at very high dilutions. PMID- 19186890 TI - The AMCP format for formulary submissions version 2.1. PMID- 19186891 TI - Editorial. AB - The Journal of Applied Psychology is the oldest and largest top-tier journal publishing theory and research relevant to industrial and organizational psychology, organizational behavior, and human resources management. The primary emphasis of this journal is the publication of original investigations that advance theoretical understanding and create new knowledge for applied psychology within the broad scope of the organizational sciences. We are primarily interested in publishing empirical research and conceptual articles that enhance understanding of psychological phenomena in human and organizational systems. This editorial also covers the expectations and review process that the Journal of Applied Psychology has for manuscripts submitted to the journal. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19186892 TI - Interaction patterns in crisis negotiations: persuasive arguments and cultural differences. AB - This research examines cultural differences in negotiators' responses to persuasive arguments in crisis (hostage) negotiations over time. Using a new method of examining cue-response patterns, the authors examined 25 crisis negotiations in which police negotiators interacted with perpetrators from low context (LC) or high-context (HC) cultures. Compared with HC perpetrators, LC perpetrators were found to use more persuasive arguments, to reciprocate persuasive arguments in the second half of negotiations, and to respond to persuasive arguments in a compromising way. Further analyses found that LC perpetrators were more likely to communicate threats, especially in the first half of the negotiations, but that HC perpetrators were more likely to reciprocate them. The implications of these findings for our understanding of intercultural interaction are discussed. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19186893 TI - The popularity contest at work: who wins, why, and what do they receive? AB - In 2 studies, the authors investigated the popularity of employees at work. They tested a model that positioned personality in the form of core self-evaluations and situational position in the form of communication network centrality as antecedents of popularity and interpersonal citizenship and counterproductive work behaviors received from coworkers as outcomes of popularity. Data from 116 employees and 383 coworkers in Study 1 and 139 employees, their significant others, and 808 coworkers in Study 2 generally supported the model. Core self evaluations and communication network centrality were positively related to popularity, and popular employees reported receiving more citizenship behaviors and fewer counterproductive work behaviors from their coworkers than less popular employees, even controlling for interpersonal liking. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19186894 TI - Sexual behavior at work: Fun or folly? AB - Sexual behavior at work (e.g., sexual jokes and propositions) has been largely portrayed as offensive and harmful. The current research represents the first studies to test whether this is typically the case. Study 1 surveyed manufacturing and social service workers (N = 238) about their psychological well being, work withdrawal, and exposure to sexual behavior at work. Respondents indicated how often they were exposed to different sexual behaviors and how much they enjoyed or were bothered by them. Study 2 surveyed university staff (N = 1,004) about their psychological well-being, drug use, feelings of being valued at work, and exposure to sexual behavior at work. Fifty-eight percent of employees in Study 1 were exposed to sexual behavior in the past 2 years; 40% of employees in Study 2 were exposed to sexual behavior in the past year. Some women and many men reported enjoying sexual behavior at work. Despite this, exposure to sexual behavior at work predicted negative employee work and psychological well being, even for employees who said they enjoyed the experience. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19186895 TI - Developing a theory of the strategic core of teams: a role composition model of team performance. AB - Although numerous models of team performance have been articulated over the past 20 years, these models have primarily focused on the individual attribute approach to team composition. The authors utilized a role composition approach, which investigates how the characteristics of a set of role holders impact team effectiveness, to develop a theory of the strategic core of teams. Their theory suggests that certain team roles are most important for team performance and that the characteristics of the role holders in the "core" of the team are more important for overall team performance. This theory was tested in 778 teams drawn from 29 years of major league baseball (1974'-2002). Results demonstrate that although high levels of experience and job-related skill are important predictors of team performance, the relationships between these constructs and team performance are significantly stronger when the characteristics are possessed by core role holders (as opposed to non-core role holders). Further, teams that invest more of their financial resources in these core roles are able to leverage such investments into significantly improved performance. These results have implications for team composition models, as they suggest a new method for considering individual contributions to a team's success that shifts the focus onto core roles. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19186896 TI - Do you see what we see? The complex effects of perceptual distance between leaders and teams. AB - Previous distance-related theories and concepts (e.g., social distance) have failed to address the sometimes wide disparity in perceptions between leaders and the teams they lead. Drawing from the extensive literature on teams, leadership, and cognitive models of social information processing, the authors develop the concept of leader-team perceptual distance, defined as differences between a leader and a team in perceptions of the same social stimulus. The authors investigate the effects of perceptual distance on team performance, operationalizing the construct with 3 distinct foci: goal accomplishment, constructive conflict, and decision-making autonomy. Analyzing leader, member, and customer survey responses for a large sample of teams, the authors demonstrate that perceptual distance between a leader and a team regarding goal accomplishment and constructive conflict have a nonlinear relationship with team performance. Greater perceptual differences are associated with decreases in team performance. Moreover, this effect is strongest when a team's perceptions are more positive than the leader's are (as opposed to the reverse). This pattern illustrates the pervasive effects that perceptions can have on team performance, highlighting the importance of developing awareness of perceptions in order to increase effectiveness. Implications for theory and practice are delineated. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19186897 TI - Managing diversity and enhancing team outcomes: the promise of transformational leadership. AB - In a sample of 62 research and development (R&D) teams, the authors examined transformational leadership as a moderator of the relationship of age, nationality, and educational background diversity with team outcomes. When levels of transformational leadership were high, nationality and educational diversity were positively related to team leaders' longitudinal ratings of team performance. These relationships were nonsignificant when transformational leadership was low. Age diversity was not related to team performance when transformational leadership was high, and it was negatively related to team performance when transformational leadership was low. Two mediated moderation effects help explain these findings. Transformational leadership moderated the relationship of the 3 examined diversity dimensions with the elaboration of task relevant information, which in turn was positively associated with team performance. Moreover, transformational leadership moderated the relationship of the 3 diversity types with collective team identification, which in turn was positively related to the elaboration of task-relevant information. The authors discuss the theoretical and practical implications of these results. Overall, this study suggests that transformational leadership can foster the utilization of the potential, but frequently untapped, benefits entailed by both demographic and informational/cognitive team diversity. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19186898 TI - Laying the foundation for successful team performance trajectories: The roles of team charters and performance strategies. AB - This study examined the influences of team charters and performance strategies on the performance trajectories of 32 teams of master's of business administration students competing in a business strategy simulation over time. The authors extended existing theory on team development by demonstrating that devoting time to laying a foundation for both teamwork (i.e., team charters) and taskwork (performance strategies) can pay dividends in terms of more effective team performance over time. Using random coefficients growth modeling techniques, they found that teams with high-quality performance strategies outperformed teams with poorer quality strategies. However, a significant interaction between quality of the charters of teams and their performance strategies was found, such that the highest sustained performances were exhibited by teams that were high on both features. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19186899 TI - Transfer of management training from alternative perspectives. AB - One hundred seven management training evaluations were meta-analyzed to compare effect sizes for the transfer of managerial training derived from different rating sources (self, superior, peer, and subordinate) and broken down by both study- and training-related variables. For studies as a whole, and interpersonal management skills training studies in particular, transfer effects based on trainees' self-ratings, and to a lesser extent ratings from their superiors, were largest and most varied across studies. In contrast, transfer effects based on peer ratings, and particularly subordinate ratings, were substantially smaller and more homogeneous. This pattern was consistent across different sources of studies, features of evaluation design, and within a subset of 14 studies that each included all 4 rating sources. Across most rating sources, transfer of training was greatest for studies conducted in nonmilitary settings, when raters were likely to have known whether the manager being rated had attended training, when criteria were targeted to training content, when training content was derived from an analysis of tasks and skill requirements, and when training included opportunities for practice. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19186900 TI - Individual- and organizational-level consequences of organizational citizenship behaviors: A meta-analysis. AB - Although one of the main reasons for the interest in organizational citizenship behaviors (OCBs) is the potential consequences of these behaviors, no study has been reported that summarizes the research regarding the relationships between OCBs and their outcomes. Therefore, the purpose of this study is to provide a meta-analytic examination of the relationships between OCBs and a variety of individual- and organizational-level outcomes. Results, based on 168 independent samples (N = 51,235 individuals), indicated that OCBs are related to a number of individual-level outcomes, including managerial ratings of employee performance, reward allocation decisions, and a variety of withdrawal-related criteria (e.g., employee turnover intentions, actual turnover, and absenteeism). In addition, OCBs were found to be related (k = 38; N = 3,611 units) to a number of organizational-level outcomes (e.g., productivity, efficiency, reduced costs, customer satisfaction, and unit-level turnover). Of interest, somewhat stronger relationships were observed between OCBs and unit-level performance measures in longitudinal studies than in cross-sectional studies, providing some evidence that OCBs are causally related to these criteria. The implications of these findings for both researchers and practitioners are discussed. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19186901 TI - When do fair procedures not matter? A test of the identity violation effect. AB - Considerable research has demonstrated that fair procedures help improve reactions to decisions, a phenomenon known as the fair process effect. However, in the present research, the authors identify when and why objectively fair procedures (i.e., receiving voice) may not always improve justice perceptions. Findings from 2 studies (Ns = 108 and 277) yield support for the proposed identity violation effect, which posits that when an outcome violates a central aspect of one's self (i.e., personal and/or social identity), objectively fair procedures do not improve procedural and distributive justice perceptions. Further, consistent with the motivated reasoning hypothesis, the Voice x Identity Violation interaction on justice perceptions was mediated by participants' tendency to find a procedural flaw--namely, to doubt that opinions were considered before making the decision. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19186902 TI - On the role of positive and negative affectivity in job performance: a meta analytic investigation. AB - Although interest regarding the role of dispositional affect in job behaviors has surged in recent years, the true magnitude of affectivity's influence remains unknown. To address this issue, the authors conducted a qualitative and quantitative review of the relationships between positive and negative affectivity (PA and NA, respectively) and various performance dimensions. A series of meta-analyses based on 57 primary studies indicated that PA and NA predicted task performance in the hypothesized directions and that the relationships were strongest for subjectively rated versus objectively rated performance. In addition, PA was related to organizational citizenship behaviors but not withdrawal behaviors, and NA was related to organizational citizenship behaviors, withdrawal behaviors, counterproductive work behaviors, and occupational injury. Mediational analyses revealed that affect operated through different mechanisms in influencing the various performance dimensions. Regression analyses documented that PA and NA uniquely predicted task performance but that extraversion and neuroticism did not, when the four were considered simultaneously. Discussion focuses on the theoretical and practical implications of these findings. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19186903 TI - The role of core self-evaluations in the coping process. AB - In 2 studies, the authors investigated whether core self-evaluations (CSE) serve as an integrative framework for understanding individual differences in coping processes. A meta-analytic review demonstrated that CSEs were associated with fewer perceived stressors, lower strain, less avoidance coping, more problem solving coping, and were not strongly related to emotion-focused coping. Consistent with the meta-analytic results, a daily diary study demonstrated that individuals with high CSE perceived fewer stressors, experienced less strain after controlling for stressors, and engaged in less avoidance coping. However, both studies demonstrated that emotional stability was uniquely related to the stress and coping process and that emotional stability moderated the relationship between stressors and strain. The discussion focuses on the distinction between depressive self-concept represented by CSE and the anxiety and worry represented by emotional stability. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19186904 TI - Effects of networking on career success: a longitudinal study. AB - Previous research has reported effects of networking, defined as building, maintaining, and using relationships, on career success. However, empirical studies have relied exclusively on concurrent or retrospective designs that rest upon strong assumptions about the causal direction of this relation and depict a static snapshot of the relation at a given point in time. This study provides a dynamic perspective on the effects of networking on career success and reports results of a longitudinal study. Networking was assessed with 6 subscales that resulted from combining measures of the facets of (a) internal versus external networking and (b) building versus maintaining versus using contacts. Objective (salary) and subjective (career satisfaction) measures of career success were obtained for 3 consecutive years. Multilevel analyses showed that networking is related to concurrent salary and that it is related to the growth rate of salary over time. Networking is also related to concurrent career satisfaction. As satisfaction remained stable over time, no effects of networking on the growth of career satisfaction were found. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19186905 TI - In the eyes of the beholder: A non-self-report measure of workplace deviance. AB - Because employees may be reluctant to admit to performing deviant acts, the authors of this study reexamined the commonly used self-report measure of workplace deviance developed by R. J. Bennett and S. L. Robinson (2000). Specifically, the self-report measure was modified into a non-self-report measure based on multiple other-reported assessments to address methodological concerns with self-reported information regarding deviant workplace behaviors. The authors assessed the psychometric properties of this new measure by first conducting an exploratory factor analysis, which indicated a 3-factor structure (production deviance, property deviance, and personal aggression). Subsequent confirmatory factor analysis on a different sample verified these findings. Taken together, the results suggest that the content and psychometric qualities of this non-self report measure of workplace deviance closely represent S. L. Robinson and R. J. Bennett's (1995) original typology of workplace deviance. The potential usefulness of this measure in organizational studies is discussed. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19186906 TI - Performance in intercultural interactions at work: cross-cultural differences in response to behavioral mirroring. AB - This article examines how performance in intercultural workplace interactions can be compromised even in the absence of overt prejudice. The authors show that individuals respond differently to nonverbal behavioral mirroring cues exhibited in workplace interactions, depending on their cultural group membership. In a field study with experienced managers, U.S. Anglos and U.S. Latinos interacted with a confederate who, unbeknownst to the participant, engaged (or not) in behavioral mirroring. Results show that the level of the confederate's mirroring differentially affected Latinos' state anxiety, but not Anglos' state anxiety, as well as actual performance in the interaction. Two additional laboratory experiments provide further evidence of the interactive relationship of behavioral mirroring and cultural group membership on evaluations of workplace interactions. Implications for intercultural interactions and research are discussed. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19186907 TI - Effects of customization on application decisions and applicant pool characteristics in a web-based recruitment context. AB - The authors examined 2 forms of customization in a Web-based recruitment context. Hypotheses were tested in a controlled study in which participants viewed multiple Web-based job postings that each included information about multiple fit categories. Results indicated that customization of information regarding person organization (PO), needs-supplies, and demands-abilities (DA) fit (fit information customization) and customization of the order in which these fit categories were presented (configural customization) had differential effects on outcomes. Specifically, (a) applicant pool PO and DA fit were greater when fit information customization was provided, (b) applicant pool fit in high- versus low-relevance fit categories was better differentiated when configural customization was provided, and (c) overall application rates were lower when either or both forms of customization were provided. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19186908 TI - Substitutes for procedural fairness: prototypical leaders are endorsed whether they are fair or not. AB - This article extends research on leader procedural fairness as well as the social identity model of leadership effectiveness (SIMOL) by demonstrating that leader prototypicality can act as a substitute for procedural fairness. Although procedural fairness in general and voice in particular have been found to have a robust positive influence on leader endorsement, the authors showed in an experimental scenario study and a correlational field study that the influence of voice on leader endorsement is substantially reduced when leaders are perceived as prototypical for the group that they lead and followers are highly identified with their group. Additionally, supportive of predictions of the SIMOL, leader prototypicality interacted with follower identification in predicting leader endorsement, such that prototypicality had a positive effect on leader endorsement, which was enhanced among high identifiers. Overall, these results suggest that leaders who are attuned to their followers' group identity can afford to go it alone, for the better or the worse. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19186909 TI - Innovation implementation in the public sector: an integration of institutional and collective dynamics. AB - The present study integrates institutional factors and employee-based collective processes as predictors of 2 key implementation outcomes: implementation effectiveness and innovation effectiveness (Klein, Conn, & Sorra, 2001). Specifically, the authors proposed that institutional factors shape employees' collective implementation efficacy and innovation acceptance. The authors further hypothesized that these employee-based collective processes mediate the effects of institutional factors on implementation outcomes. This integrative framework was examined in the context of 47 agencies and ministries of the Korean Government that were implementing a process innovation called E-Government. Three wave longitudinal data were collected from 60 external experts and 1,732 government employees. The results reveal the importance of management support for collective implementation efficacy, which affected employees' collective acceptance of the innovation. As hypothesized, these collective employee dynamics mediated the effects of institutional enablers on successful implementation as well as the amount of long-term benefit that accrued to the agencies and ministries. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19186910 TI - Work stressors, role-based performance, and the moderating influence of organizational support. AB - As a test of the 2-dimensional model of work stressors, the present study proposed differential relationships between challenge stressors and hindrance stressors and role-based performance, which were expected to be moderated by organizational support. In a sample of 215 employees across 61 offices of a state agency, the authors obtained a positive relationship between challenge stressors and role-based performance and a negative relationship between hindrance stressors and role-based performance. In addition, organizational support moderated the relationship between challenge stressors and role-based performance but did not moderate the relationship between hindrance stressors and role-based performance. This suggests that organizations would benefit from increasing challenges in the workplace as long as they are supportive of employees and removing hindrances. Further implications for organizational theory and practice are discussed. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19186911 TI - Human resources management and firm performance: The differential role of managerial affective and continuance commitment. AB - In this study, the authors developed a dual-concern (i.e., maintenance and performance) model of human resources (HR) management. The authors identified commonly examined HR practices that apply to the middle manager level and classified them into the maintenance- and performance-oriented HR subsystems. The authors found support for the 2-factor model on the basis of responses from 2,148 managers from 463 firms operating in China. Regression results indicate that the performance-oriented HR subsystems had a positive relationship with firm performance and that the relationship was mediated by middle managers' affective commitment to the firm. The maintenance-oriented HR subsystems had a positive relationship with middle managers' continuance commitment but not with their affective commitment and firm performance. This study contributes to the understanding of how HR practices relate to firm performance and offers an improved test of the argument that valuable and firm-specific HR provide a source of competitive advantage. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19186912 TI - What you feel is how you compare: how comparisons influence the social induction of affect. AB - Concordant and discordant affective reactions can occur after the mere perception of another person's affective expression. Most previous theorizing has been concerned with the explanation of affective concordance, typically referred to as emotional contagion, although discordant affect has received little attention. The authors propose an integrative account for the explanation of both outcomes based on a social comparison framework. Studies 1 and 2 suggest that two distinct types of comparison processes can trigger concordant or discordant affective reactions. Study 3 extends these findings by demonstrating that the influence of comparison processes on affect in an established mood contagion paradigm. The authors attempt to integrate previous research into the present account. PMID- 19186913 TI - Positive and negative emotionality: trajectories across six years and relations with social competence. AB - The goals of the present study were to examine (1) the mean-level stability and differential stability of children's positive emotional intensity, negative emotional intensity, expressivity, and social competence from early elementary school-aged to early adolescence, and (2) the associations between the trajectories of children's emotionality and social functioning. Using four waves of longitudinal data (with assessments 2 years apart), parents and teachers of children (199 kindergarten through third grade children at the first assessment) rated children's emotion-related responding and social competence. For all constructs, there was evidence of mean-level decline with age and stability in individual differences in rank ordering. Based on age-centered growth-to-growth curve analyses, the results indicated that children who had a higher initial status on positive emotional intensity, negative emotional intensity, and expressivity had a steeper decline in their social skills across time. These findings provide insight into the stability and association of emotion-related constructs to social competence across the elementary and middle school years. PMID- 19186914 TI - Visual search for facial expressions of emotions: a comparison of dynamic and static faces. AB - A number of past studies have used the visual search paradigm to examine whether certain aspects of emotional faces are processed preattentively and can thus be used to guide attention. All these studies presented static depictions of facial prototypes. Emotional expressions conveyed by the movement patterns of the face have never been examined for their preattentive effect. The present study presented for the first time dynamic facial expressions in a visual search paradigm. Experiment 1 revealed efficient search for a dynamic angry face among dynamic friendly faces, but inefficient search in a control condition with static faces. Experiments 2 to 4 suggested that this pattern of results is due to a stronger movement signal in the angry than in the friendly face: No (strong) advantage of dynamic over static faces is revealed when the degree of movement is controlled. These results show that dynamic information can be efficiently utilized in visual search for facial expressions. However, these results do not generally support the hypothesis that emotion-specific movement patterns are always preattentively discriminated. PMID- 19186915 TI - The look of fear and anger: facial maturity modulates recognition of fearful and angry expressions. AB - The current series of studies provide converging evidence that facial expressions of fear and anger may have co-evolved to mimic mature and babyish faces in order to enhance their communicative signal. In Studies 1 and 2, fearful and angry facial expressions were manipulated to have enhanced babyish features (larger eyes) or enhanced mature features (smaller eyes) and in the context of a speeded categorization task in Study 1 and a visual noise paradigm in Study 2, results indicated that larger eyes facilitated the recognition of fearful facial expressions, while smaller eyes facilitated the recognition of angry facial expressions. Study 3 manipulated facial roundness, a stable structure that does not vary systematically with expressions, and found that congruency between maturity and expression (narrow face-anger; round face-fear) facilitated expression recognition accuracy. Results are discussed as representing a broad co evolutionary relationship between facial maturity and fearful and angry facial expressions. PMID- 19186916 TI - When hunger finds no fault with moldy corn: food deprivation reduces food-related disgust. AB - The main purpose of this study was to examine if disgust toward unpalatable foods would be reduced among food-deprived subjects and if this attenuation would occur automatically even under moderate levels of food deprivation. Subjects were either satiated or food deprived for 15 hours and electromyographic activity was recorded at the levator muscle region while they were watching pictures of palatable versus unpalatable foods, and pictures of positive versus disgust related control pictures. For control purposes, subjects' activity of the zygomaticus and corrugator muscles was also recorded. As compared with satiated subjects, food-deprived subjects exhibited stronger activity in the zygomaticus muscle region when watching pictures of palatable foods (but not when watching positive control pictures). More important, hungry subjects exhibited weaker activity in the levator muscle region when watching pictures of unpalatable foods (but not when watching disgusting control pictures). Thus, this is the first study ever to show that specific emotions (disgust) are moderated by homeostatic dysregulation automatically. Results indicate that the modulation of facial expressions might play an important role in lowering the threshold for food intake. PMID- 19186918 TI - Behavioral facilitation: a cognitive model of individual differences in approach motivation. AB - Approach motivation consists of the active, engaged pursuit of one's goals. The purpose of the present three studies (N = 258) was to examine whether approach motivation could be cognitively modeled, thereby providing process-based insights into personality functioning. Behavioral facilitation was assessed in terms of faster (or facilitated) reaction time with practice. As hypothesized, such tendencies predicted higher levels of approach motivation, higher levels of positive affect, and lower levels of depressive symptoms and did so across cognitive, behavioral, self-reported, and peer-reported outcomes. Tendencies toward behavioral facilitation, on the other hand, did not correlate with self reported traits (Study 1) and did not predict avoidance motivation or negative affect (all studies). The results indicate a systematic relationship between behavioral facilitation in cognitive tasks and approach motivation in daily life. Results are discussed in terms of the benefits of modeling the cognitive processes hypothesized to underlie individual differences motivation, affect, and depression. PMID- 19186919 TI - Predicting the duration of emotional experience: two experience sampling studies. AB - The authors present 2 studies to explain the variability in the duration of emotional experience. Participants were asked to report the duration of their fear, anger, joy, gratitude, and sadness episodes on a daily basis. Information was further collected with regard to potential predictor variables at 3 levels: trait predictors, episode predictors, and moment predictors. Discrete-time survival analyses revealed that, for all 5 emotions under study, the higher the importance of the emotion-eliciting situation and the higher the intensity of the emotion at onset, the longer the emotional experience lasts. Moreover, a reappearance, either physically or merely mentally, of the eliciting stimulus during the emotional episode extended the duration of the emotional experience as well. These findings display interesting links with predictions within N. H. Frijda's theory of emotion, with the phenomenon of reinstatement (as studied within the domain of learning psychology), and with the literature on rumination. PMID- 19186917 TI - Individual differences in valence modulation of face-selective M170 response. AB - Magnetoencephalography was used to examine the effect of individual differences on the temporal dynamics of emotional face processing by grouping subjects based on their ability to detect masked valence-laden stimuli. Receiver operating characteristic curves and a nonparametric sensitivity measure were used to categorize subjects into those that could and could not reliably detect briefly presented fearful faces that were backward-masked by neutral faces. Results showed that, in a cluster of face-responsive sensors, the strength of the M170 response was modulated by valence only when subjects could reliably detect the masked fearful faces. Source localization of the M170 peak using synthetic aperture magnetometry identified sources in face processing areas such as right middle occipital gyrus and left fusiform gyrus that showed the valence effect for those target durations at which subjects were sensitive to the fearful stimulus. Subjects who were better able to detect fearful faces also showed higher trait anxiety levels. These results suggest that individual differences between subjects, such as trait anxiety levels and sensitivity to fearful stimuli, may be an important factor to consider when studying emotion processing. PMID- 19186920 TI - When frustration is repeated: behavioral and emotion responses during extinction over time. AB - Persistence of instrumental responding and negative facial expressions in response to repeated goal blockage was studied in 53 4-month-old infants. All participants experienced 2 sessions comprising baseline (no stimulation), contingency (stimulation resulting from infant action), and extinction (no stimulation) on consecutive days. Performance criteria identified 2 groups of infants, those who learned in Session 1 (Learning Group 1) and those who learned in Session 2 (Learning Group 2). Individual differences in instrumental responses and facial expression during extinction were compared as a function of learning group. Across sessions, the repetition of extinction for Learning Group 1 was associated with both a persistent instrumental response and anger expressions. The level of instrumental response and anger expression was equivalent to that observed for Learning Group 2 but only in Session 2, the day on which that group learned. Sadness and anger/sadness blended expressions were initially more common in Learning Group 2, but these expressions were attenuated given another exposure to the contingency in Session 2. Implications for the relations among infant emotion, cognition, and behavior are discussed. PMID- 19186921 TI - Willingness to express emotions to caregiving spouses. AB - This study examined the association between care-recipients' willingness to express emotions to spousal caregivers and caregiver's well-being and support behaviors. Using self-report measures in the context of a larger study, 262 care recipients with osteoarthritis reported on their willingness to express emotions to caregivers, and caregivers reported on their stress and insensitive responding to care-recipients. Results revealed that care-recipients' willingness to express happiness was associated with less insensitive caregiver responding, and willingness to express interpersonal emotions (e.g., compassion, guilt) was associated with less caregiving stress. There were also gender differences, such that caregiving wives, in particular, benefited from their husband's willingness to express vulnerable (e.g., anxiety, sadness) and interpersonal emotions. PMID- 19186922 TI - Children's extension of disgust to physical and moral events. AB - Adults use the terms revolting, gross, and disgusting to describe entities and actions, such as feces, rotten food, and sex with corpses, which elicit a certain visceral response. But adults also apply such expressions to certain sociomoral transgressions, such as cheating on one's spouse or stealing from the poor. Here, the authors explore whether young children associate disgust with physical and moral events by endorsing either verbal or facial expressions of disgust. Results indicate that children in Grades K, 2, and 4 (N = 167) label moral violations "disgusting" more often than nondisgusting physical acts or neutral negative acts but less often than physically disgusting acts. Likewise, children associate facial expressions of disgust with moral violations. These findings are discussed in the context of different theories about the relationship between physical disgust and moral disgust. PMID- 19186923 TI - Jealousy: novel methods and neural correlates. AB - Because of the difficulties surrounding the evocation of jealousy, past research has relied on reactions to hypothetical scenarios and recall of past experiences of jealousy. Both methodologies have limitations, however. The present research was designed to develop a method of evoking jealousy in the laboratory that would be well controlled, ethically permissible, and psychologically meaningful. Study 1 demonstrated that jealousy could be evoked in a modified version of K. D. Williams' (2007) Cyberball ostracism paradigm in which the rejecting person was computer-generated. Study 2, the first to examine neural activity during the active experience of jealousy, tested whether experienced jealousy was associated with greater relative left or right frontal cortical activation. The findings revealed that the experience of jealousy was correlated with greater relative left frontal cortical activation toward the "sexually" desired partner. This pattern of activation suggests that jealousy is associated with approach motivation. Taken together, the present studies developed a laboratory paradigm for the study of jealousy that should help foster research on one of the most social of emotions. PMID- 19186924 TI - When guilt evokes self-punishment: evidence for the existence of a Dobby Effect. AB - Feelings of guilt may be resolved in various ways. The scientific literature has mainly highlighted beneficial interpersonal consequences of guilt, showing repeatedly that guilt motivates compensatory pro-social behavior to repair social bonds. The authors reveal that when opportunities for compensation are not present, guilt may evoke self-punishment. Self-punishment was demonstrated through self-denied pleasure in a scenario study, and by self-enforced penalties in an experimental study. The authors call this tendency for self-punishment the Dobby Effect, and discuss it as an explanation for the widely held conviction that atonement absolves sins, its contribution to some types of psychopathology, as well as its possible functional relevance. PMID- 19186925 TI - The feeling of uncertainty intensifies affective reactions. AB - Uncertainty has been defined as a lack of information about an event and has been characterized as an aversive state that people are motivated to reduce. The authors propose an uncertainty intensification hypothesis, whereby uncertainty during an emotional event makes unpleasant events more unpleasant and pleasant events more pleasant. The authors hypothesized that this would happen even when uncertainty is limited to the feeling of "not knowing," separable from a lack of information. In 4 studies, the authors held information about positive and negative film clips constant while varying the feeling of not knowing by having people repeat phrases connoting certainty or uncertainty while watching the films. As predicted, the subjective feeling of uncertainty intensified people's affective reactions to the film clips. PMID- 19186926 TI - Shared perceptual basis of emotional expressions and trustworthiness impressions from faces. AB - Using a dynamic stimuli paradigm, in which faces expressed either happiness or anger, the authors tested the hypothesis that perceptions of trustworthiness are related to these expressions. Although the same emotional intensity was added to both trustworthy and untrustworthy faces, trustworthy faces who expressed happiness were perceived as happier than untrustworthy faces, and untrustworthy faces who expressed anger were perceived as angrier than trustworthy faces. The authors also manipulated changes in face trustworthiness simultaneously with the change in expression. Whereas transitions in face trustworthiness in the direction of the expressed emotion (e.g., high-to-low trustworthiness and anger) increased the perceived intensity of the emotion, transitions in the opposite direction decreased this intensity. For example, changes from high to low trustworthiness increased the intensity of perceived anger but decreased the intensity of perceived happiness. These findings support the hypothesis that changes along the trustworthiness dimension correspond to subtle changes resembling expressions signaling whether the person displaying the emotion should be avoided or approached. PMID- 19186927 TI - Conditioning-induced attentional bias for face stimuli measured with the emotional Stroop task. AB - People with anxiety disorder display attentional bias toward threat-related objects. Using classical fear conditioning, the authors investigated the possible source of such bias in normal participants. Following differential fear conditioning in which an angry face of either male or female (conditioned stimulus: CS+) was paired with mild electric fingershock (unconditioned stimulus: US) but the angry face of the other gender and all other facial expressions unpaired (CS-), an emotional Stroop task was administered. In the Stroop task, participants were required to identify the color of the facial stimuli (red, green, blue, or yellow). Response latency was significantly longer for the CS+ angry face than the other unpaired facial stimuli (CS-). Furthermore, this acquired attentional bias was positively correlated with the level of trait anxiety measured before the conditioning and the degree of self-reported aversiveness of the US. Our results demonstrated that attentional bias could be induced in normal individuals through a simple associative learning procedure, and the acquisition is modulated by the level of trait anxiety and the level of perceived fear of the aversive US. PMID- 19186928 TI - Attentional control of emotional distraction in rapid serial visual presentation. AB - Temperament research has highlighted the importance of attentional control in both emotion regulation and as a predictor of psychopathology. Enhanced susceptibility to emotional distraction is a key feature of mood disturbance. Whereas many studies have examined the influence of individual differences in anxiety on the disruptive effects of emotional distractors, individual differences in attentional control have been largely neglected. Here we examine, within healthy volunteers, the relative contributions of individual differences in self-reported anxiety and attentional control to distractibility caused by emotional or neutral faces distractors occurring prior to neutral face targets during rapid serial visual presentation. Participants with good attentional control were less affected by both neutral and emotional distractors than participants with poorer attentional control. More pronounced distraction deficits were seen for emotional relative to neutral distractors in individuals with poor attentional control. In contrast state anxiety was not associated with increased emotional distraction. Our findings suggest a protective role of attentional control mechanisms in minimizing the influence of emotional distraction. PMID- 19186929 TI - Alcohol expectancies and reactivity to alcohol-related and affective cues. AB - Recent conceptualizations of alcohol expectancies relate cognitive schemas to their neurobiological underpinnings; cue reactivity paradigms lend themselves well to testing this broadened conceptual framework. In the present study, we examined the relationship between self-reported alcohol expectancies and responses to alcohol-related and affective picture cues among fifty-five young adults. In addition to traditional subjective and psychophysiological indices of cue reactivity, the startle eyeblink reflex was obtained during picture cue presentations to address both attention-arousal (early probes) and affective motivational (late probes) aspects of cue processing. Analyses indicated that participants reporting greater positive, arousing, and social alcohol expectancies rated alcohol cues as more pleasant, arousing, and craving-inducing. In addition, participants displayed inhibited startle reactivity to late alcohol cue probes, indicative of an appetitive reaction. Finally, startle responding to early probes indicated that participants with greater alcohol expectancies displayed blunted attention to negative affect cues. Findings are discussed in terms of the utility of the startle reflex and cue reactivity paradigms for clarifying the relationship between alcohol expectancies and motivated attention to salient cues. PMID- 19186930 TI - Methamphetamine impairs sexual motivation but not sexual performance in male Japanese quail. AB - The present study investigated the effects of chronic pre-exposure to methamphetamine on sexual motivation and performance in male Japanese quail. Quail were pre-exposed to methamphetamine (1.0 or 3.0 mg/kg ip) or saline (ip) once daily for 10 days and locomotor activity was measured. After a 10 day washout period, sexual motivation was measured in a straight-arm runway with visual access to a female at one end. Three to 5 hr after sexual motivation tests, males were allowed to copulate with a receptive female quail and copulatory behavior was assessed. Tests were conducted once per day for 10 days. Results showed that males pre-exposed to methamphetamine had decreased locomotor activity compared to saline controls. Males pre-exposed to METH later ran slower toward a female in the runway and spent less time near her. In contrast, methamphetamine pre-exposed males showed similar copulatory behavior as saline pre-exposed males. The findings suggest that chronic pre-exposure to methamphetamine may impair sexual motivation but not sexual performance. The findings are discussed from a comparative perspective and with regard to their clinical relevance. PMID- 19186931 TI - Detection versus sustained attention to drug cues have dissociable roles in mediating drug seeking behavior. AB - It is commonly thought that attentional bias for drug cues plays an important role in motivating human drug-seeking behavior. To assess this claim, two groups of smokers were trained in a discrimination task in which a tobacco-seeking response was rewarded only in the presence of 1 particular stimulus (the S+). The key manipulation was that whereas 1 group could control the duration of S+ presentation, for the second group, this duration was fixed. The results showed that the fixed-duration group acquired a sustained attentional bias to the S+ over training, indexed by greater dwell time and fixation count, which emerged in parallel with the control exerted by the S+ over tobacco-seeking behavior. By contrast, the controllable-duration group acquired no sustained attentional bias for S+ and instead used efficient detection of the S+ to achieve a comparable level of control over tobacco seeking. These data suggest that detection and sustained attention to drug cues have dissociable roles in enabling drug cues to motivate drug-seeking behavior, which has implications for attentional retraining as a treatment for addiction. PMID- 19186932 TI - Relations between anxiety sensitivity, distress tolerance, and fear reactivity to bodily sensations to coping and conformity marijuana use motives among young adult marijuana users. AB - The present investigation examines anxiety sensitivity, distress tolerance, and fear reactivity to bodily sensations in relation to Coping and Conformity marijuana use motives among a sample of young adult marijuana users (n = 135; 46.7% women; Mage = 20.45, SD = 5.0). After controlling for current marijuana use frequency (past 30 days), daily cigarette smoking rate, average volume of alcohol used over the past year, negative affectivity, and other marijuana use motives, anxiety sensitivity was significantly and uniquely associated with Coping and Conformity motives for marijuana use. Distress tolerance evidenced significant and unique incremental relations to Coping motives, whereas fear reactivity to bodily sensations was unrelated to any marijuana use motive. These results provide novel information related to the role of emotional sensitivity and tolerance factors as they pertain to specific types of motives for marijuana use among young adults. PMID- 19186933 TI - Competing visual and olfactory imagery tasks suppress craving for coffee. AB - Using the elaborated intrusion theory of desire as a theoretical basis, the present experiments investigated whether mutual competition between modality specific tasks and desire-related imagery can suppress cravings for caffeine--in particular, coffee. Data from a pilot survey (Experiment 1) in 106 female undergraduate habitual coffee drinkers showed that coffee cravings are characterized predominantly by sensory images in the visual, olfactory, and gustatory modalities. Experiments 2 and 3 tested the prediction that a competing visual or olfactory imagery task would reduce such cravings, relative to an auditory imagery task. In each experiment, 90 female undergraduate habitual coffee drinkers were asked to forego their morning cup of coffee and were subsequently exposed to pictures of coffee (Experiment 2) or actual coffee (Experiment 3). After the craving induction protocol, participants completed either a visual, auditory, or olfactory imagery task with neutral (i.e., non coffee-related) content. As predicted, the visual and olfactory imagery tasks reduced participants' cravings for coffee, whereas the auditory task did not. Imagery techniques in the visual or olfactory domain could provide a useful adjunct to the behavioral regime of caffeine fading for consumers who are trying to quit caffeine use or reduce their intake. PMID- 19186934 TI - Disinhibitory psychopathology and delay discounting in alcohol dependence: personality and cognitive correlates. AB - Increased discounting of delayed rewards may reflect a decision bias that contributes to excessive use of alcohol and more generally, to an impulsive, disinhibitory predisposition that is characterized by a preference for immediate over long-term rewards. The current study examined the association between delay discounting of rewards and the covariation among several types of disinhibitory problems that are often comorbid with alcohol dependence (AD). Lifetime problems with alcohol, marijuana, other drugs, childhood conduct disorder, and adult antisocial behavior were assessed in a sample of 426 young adults, 257 of whom had a lifetime diagnosis of AD. Higher delay discounting rates were associated with the covariation among all domains of disinhibitory problems and were not uniquely associated with any one domain. Higher delay discounting rates also were associated with lower intelligence, lower working memory capacity, and higher trait impulsivity. The results suggest that increased delay discounting of rewards may reflect aspects of a general vulnerability to externalizing, disinhibitory disorders. PMID- 19186935 TI - Standoff spectroscopy of surface adsorbed chemicals. AB - Despite its immediate applications, selective detection of trace quantities of surface adsorbed chemicals, such as explosives, without physically collecting the sample molecules is a challenging task. Standoff spectroscopic techniques offer an ideal method of detecting chemicals without using a sample collection step. Though standoff spectroscopic techniques are capable of providing high selectivity, their demonstrated sensitivities are poor. Here we describe standoff detection of trace quantities of surface adsorbed chemicals using two quantum cascade lasers operated simultaneously, with tunable wavelength windows that match with absorption peaks of the analytes. This standoff method is a variation of photoacoustic spectroscopy, where scattered light from the sample surface is used for exciting acoustic resonance of the detector. We demonstrate a sensitivity of 100 ng/cm(2) and a standoff detection distance of 20 m for surface adsorbed analytes such as explosives and tributyl phosphate. PMID- 19186936 TI - Highly efficient enzyme reactors containing trypsin and endoproteinase LysC immobilized on porous polymer monolith coupled to MS suitable for analysis of antibodies. AB - Capillary enzymatic microreactors containing trypsin and endoproteinase LysC immobilized on a porous polymer monolith have been prepared and used for the characterization and identification of proteins such as cytochrome c, bovine serum albumin, and high-molecular weight human immunoglobulin G. The hydrophilicity of diol functionalities originating from the hydrolyzed poly(glycidyl methacrylate-co-ethylene dimethacrylate) monolith was not sufficient to avoid adsorption of hydrophobic albumin in a highly aqueous mobile phase. Therefore, this monolith was first hydrophilized via photografting of poly(ethylene glycol) methacrylate followed by photografting of a 4-vinyl-2,2 dimethylazlactone to provide the pore surface with reactive functionalities required for immobilization. This new approach reduced the undesired nonspecific adsorption of proteins and peptides and facilitated control of both the enzyme immobilization and protein digestion processes. The enzymatic reactors were coupled off-line with MALDI/TOF MS and/or on-line with ESI/TOF MS. Experimental conditions for digestion were optimized using cytochrome c and bovine serum albumin as model proteins. The optimized reactors were then integrated into a multidimensional system comprised of a monolithic capillary enzyme reactor, an in line nanoLC separation of peptides using a poly(lauryl methacrylate-co-ethylene dimethacrylate) monolithic column, and ESI/TOF MS. With the use of this system, immunoglobulin G was digested at room temperature in 6 min to an extent similar to that achieved with soluble enzyme at 37 degrees C after 24 h. PMID- 19186937 TI - Transglutaminase-mediated PEGylation of proteins: direct identification of the sites of protein modification by mass spectrometry using a novel monodisperse PEG. AB - Poly(ethylene glycol) (PEG) has been widely used to prolong the residence time of proteins in blood and to decrease their immunogenicity and antigenicity. A drawback of this polymer lies in its polydispersity that makes difficult the identification of the sites of protein modification. This is a mandatory requirement if a PEGylated protein should be approved as a drug. Here, a fast and reliable method is proposed to characterize proteins conjugated at the level of glutamine (Gln) residues using microbial transglutaminase (TGase). The novelty resides in the use of a monodisperse Boc-PEG-NH(2) for the derivatization that allows the direct identification of the sites of PEGylation by electrospray ionization mass spectrometry (ESI-MS). The procedure has been tested on three model proteins, namely, human granulocyte colony-stimulating factor, human growth hormone, and horse heart apomyoglobin. The Gln residues linked to the polymer chain were easily identified by ESI-MS and tandem MS analyses, demonstrating the advantage of using a monodisperse polymer in combination with mass spectrometry for an easy characterization of conjugated proteins. Interestingly, the PEGylation reaction led to the production only of mono- and bis-derivative products, indicating that the TGase-mediated PEGylation can be extremely selective and thus very useful for the derivatization of protein drugs. PMID- 19186938 TI - Cellulose/DNA hybrid nanomaterials. AB - Cellulose nanocrystals (CNXLs) have drawn attention from researchers for their remarkable reinforcing abilities and excellent mechanical properties. CNXLs typically have high aspect ratios of around 20-50 (length/width), low density of around 1.6 g/cc, high stiffness (135 to 155 GPa), and strength (estimated at 7500 MPa). Here we utilize CNXLs in a bottom-up hierarchical assembly to produce a macroscale material. Single-stranded oligonucleotides with an amino modifier were successfully grafted on CNXLs. The molecular recognition ability of the oligomeric base pairs was then utilized by duplexing complementary oligonucleotides grafted onto separate CNXL populations. The resulting hybrid nanomaterials were analyzed using dynamic light scattering, atomic force microscopy, and UV spectroscopy. PMID- 19186939 TI - Identification of microtubule-binding domains on microtubule-associated proteins by major coat phage display technique. AB - Microtubule is an important structural and functional component in cells. Microtubule-associated proteins (MAPs) are a class of proteins that can bind to microtubules and stabilize them to maintain their functions. However, not all the specific microtubule-binding domains on MAPs are clear. Here we report the study of microtubule-binding domains on MAPs from a new angle by biopanning a new type of phage-displayed random peptide library (called landscape phage library) against purified alpha- and beta-tubulins. In the landscape phage library, billions of fd-tet phage clones are present and a unique 9-mer peptide is fused to each of the approximately 3900 copies of major coat protein (pVIII) in each clone. The affinity-selected peptides derived from the biopanning were analyzed by the receptor ligand contacts (RELIC) suite of programs, which is a bioinformatics tool for combinatorial peptide analysis and identification of protein-ligand interaction sites. By using RELIC, the affinity-selected peptides were shown to have similarity with the sequences of two MAP families (MAP1 and MAP2/tau), thereby identifying putative microtubule-binding domains on these MAPs. The tubulin-binding affinity was also confirmed by using transmission electron microscopy (TEM) to characterize the interaction between affinity selected tubulin-binding phage and tubulins. Our results confirm some known microtubule-binding domains and identify some new microtubule-binding domains and thus shed light into the mechanism of microtubule-MAPs interactions. PMID- 19186940 TI - Coaxial MnO2/carbon nanotube array electrodes for high-performance lithium batteries. AB - Coaxial manganese oxide/carbon nanotube (CNT) arrays deposited inside porous alumina templates were used as cathodes in a lithium battery. Excellent cyclic stability and capacity of MnO2/CNT coaxial nanotube electrodes resulted from the hybrid nature of the electrodes with improved electronic conductivity and dual mechanism of lithium storage. The reversible capacity of the battery was increased by an order compared to template grown MnO2 nanotubes, making them suitable electrodes for advanced Li ion batteries. PMID- 19186944 TI - Ring contraction of 1,2,4,5,7,8-hexaoxa-3-silonanes by selective reduction of COOSi fragments. Synthesis of new silicon-containing rings, 1,3,5,6-tetraoxa-2 silepanes. AB - The reducing agents Ph(3)P, (C(8)H(17))(3)P, or NH(2)C(S)NH(2) promote the ring contraction of nine-membered triperoxides, viz., 1,2,4,5,7,8-hexaoxa-3-silonanes, giving rise to seven-membered rings belonging to the previously unknown class of monoperoxides, viz., 1,3,5,6-tetraoxa-2-silepanes, in yields from 67% to 91%. Therefore, the selective reduction of the SiOOC fragments to SiOC in molecules containing simultaneously the COOC fragment was performed for the first time. PMID- 19186946 TI - Malaria-infected mice are cured by a single oral dose of new dimeric trioxane sulfones which are also selectively and powerfully cytotoxic to cancer cells. AB - A new series of 6 dimeric trioxane sulfones has been prepared from the natural trioxane artemisinin in five or six chemical steps. One of these thermally and hydrolytically stable new chemical entities (4c) completely cured malaria infected mice via a single oral dose of 144 mg/kg. At a much lower single oral dose of only 54 mg/kg combined with 13 mg/kg of mefloquine hydrochloride, this trioxane dimer 4c as well as its parent trioxane dimer 4b also completely cured malaria-infected mice. Both dimers 4c and 4b were potently and selectively cytotoxic toward five cancer cell lines. PMID- 19186947 TI - Cysteine peptidase expression in Trichomonas vaginalis isolates displaying high- and low-virulence phenotypes. AB - In the present study, we identified and characterized the cysteine peptidase (CP) profiles of Trichomonas vaginalis isolates exhibiting high- and low-virulence phenotypes using a combination of two-dimensional SDS-PAGE (2DE), tandem mass spectrometry (MS/MS), and data mining. Seven of the eight CPs identified belong to Clan CA, family C1, cathepsin L-like CP, and one belongs to Clan CD, family C13, asparaginyl endopeptidase-like CP. Quantitative and qualitative differences in CP expression were detected between the isolates. BLAST analysis followed by CLUSTAL alignment of amino acid sequences of differentially expressed CPs showed identity or high homology to previously described CP cDNA clones CP1, CP3, CP4, and to a secreted CP fraction of 30 kDa involved in apoptosis of vaginal epithelial cells. One- and two-dimensional-substrate gel analyses revealed the differential CP profiles between the isolates, indicating that the combination of zymography with 2DE and MS/MS might be a powerful experimental approach to map and identify active peptidases in T. vaginalis. Toxicity exerted upon HeLa cells by high- and low-virulence isolates was 98.3% and 31%, respectively. Pretreatment of parasites with specific Clan CA papain-like CP inhibitor l-3-carboxy-2,3-trans epoxypropionyl-leucylamido(4-guanidino)butane (E-64) drastically reduced the cytotoxic effect to 21.7% and 0.8%, respectively, suggesting that T. vaginalis papain-like CPs are the main factors involved in the cellular damage. PMID- 19186948 TI - Identification of S100A8 and S100A9 as serological markers for colorectal cancer. AB - In search of novel serological protein biomarkers for human colorectal cancer (CRC), we analyzed CRC tissues using two-dimensional difference in-gel electrophoresis (2D-DIGE) on a narrow range IPG strip (pH 5.5-6.7). By comparing tumor tissues with matched normal tissues in a pairwise manner (n = 6), we identified 34 up-regulated and 17 down-regulated spots with intensity changes greater than 2-fold (Student's t-test, p < 0.05). Expression of both mRNA and protein levels of four proteins, adenosylhomocysteinase, Nm23-H1, S100A8 and S100A9, in CRC tissues was further evaluated by semiquantitative RT-PCR and Western blot analysis. The results revealed that all four proteins were elevated in the tumor tissues. We also confirmed, by immunohistochemistry, that adenosylhomocysteinase and Nm23-H1 were overexpressed in tumor cell cytoplasm and that S100A8 and S100A9 proteins were strongly expressed in tumor infiltrating immune cells. Western blot analysis with fractionated plasma samples showed that S100A8 and S100A9 were significantly increased in the plasma of CRC patients (n = 77) and colorectal adenoma patients (n = 11), compared to healthy controls (n = 21). The area under a receiver operating characteristic (ROC) curve was 0.91 for S100A8 and 0.89 for S100A9, which was superior to the established tumor marker carcinoembryonic antigen with 0.78 for the area under the ROC curve. Some patients with inflammatory diseases such as pancreatitis also showed elevated levels of the proteins. Importantly, in comparison to the control group, both proteins showed a remarkable change at the early stage of cancer. Therefore, we suggest S100A8 and S100A9 as candidates for serological biomarkers in combination with other serum markers that aid CRC diagnosis. PMID- 19186949 TI - Use of differential isotopic labeling and mass spectrometry to analyze capacitation-associated changes in the phosphorylation status of mouse sperm proteins. AB - Mammalian sperm need to reside in the female reproductive tract for a finite period of time before acquiring fertilizing competence. The biochemical changes associated with this process are collectively known as "capacitation". With the use of the mouse as an experimental model, we have previously demonstrated that capacitation is associated with a cAMP-dependent increase in protein tyrosine phosphorylation. However, little is known about the identity and function of the protein targets of this phosphorylation cascade. In the present work, we have used differential isotopic labeling coupled with immobilized metal affinity chromatography (IMAC)-based phosphopeptide enrichment and analysis on a hybrid linear ion trap/FT-ICR mass spectrometer to measure the changes in protein phosphorylation resulting from the capacitation process. As no kinase activators and/or phosphatase inhibitors were used in the preparation of the sperm samples, phosphorylated residues identified in this study represent in vivo sites of phosphorylation. Also, in contrast to other methods which rely on the incorporation of isotopically labeled amino acids at the protein level (e.g., SILAC), the present technique is based on the Fisher esterification of protein digests, allowing for the comparison of phosphorylation status in the absence of protein synthesis. This approach resulted in the identification of 55 unique, in vivo sites of phosphorylation and permitted the relative extent of phosphorylation, as a consequence of capacitation, to be calculated for 42 different phosphopeptides. This work represents the first effort to determine which specific protein phosphorylation sites change their phosphorylation status in vivo as a result of the mammalian capacitation process. PMID- 19186950 TI - Wet-spinning of recombinant silk-elastin-like protein polymer fibers with high tensile strength and high deformability. AB - A recombinant silk-elastin-like protein copolymer SELP-47K containing tandemly repeated amino acid sequence blocks from silk, GAGAGS, and elastin, GVGVP, was fabricated into microdiameter fibers using a wet-spinning technique. Raman spectral analysis revealed the formation of antiparallel beta-sheet crystals of the silk-like blocks. Dry SELP-47K fibers display the dependence of mechanical properties such as Young's modulus on fiber diameter, suggesting more oriented and crystallized molecular chains in small-diameter fibers. Additionally, a brittle fracture mode was identified for dry fibers by SEM analysis of fracture surfaces. Hydration dramatically influenced the mechanical behavior of SELP-47K fibers. In contrast to the high tensile strength and limited strains to failure of dry fibers, fully hydrated SELP-47K fibers possessed strains to failure as high as 700%. Furthermore, upon chemical cross-linking, a tensile mechanical strength up to 20 MPa was achieved in hydrated fibers without compromising their high deformability. By combing the silk- and elastin-derived sequences into a single SELP-47K protein polymer, we demonstrated that protein fibers with high tensile strength and high deformability can be fabricated. PMID- 19186951 TI - Structural characterization of bioengineered alpha-D-glucans produced by mutant glucansucrase GTF180 enzymes of Lactobacillus reuteri strain 180. AB - Mutagenesis of specific amino acid residues of the glucansucrase (GTF180) enzyme from Lactobacillus reuteri strain 180 yielded 12 mutant enzymes that produced modified exopolysaccharides (mEPSs) from sucrose. Ethanol-precipitated and purified mEPSs were subjected to linkage analysis, Smith degradation analysis, and 1D/2D (1)H NMR spectroscopy. Comparison of the results with structural data of the previously described wild type EPS180 and triple mutant mEPS-PNNS revealed a broad variation of structural elements between mEPS molecules. The amount of (alpha1-->3) linkages varied from 14-43%, the amount of (alpha1-->4) linkages (not present in the wild type) from 0-12%, and the amount of (alpha1-->6) linkages from 51-86%. The average molecular weight (M(w)) ranged from 9.4 to 32.3 MDa and the degree of branching varied from 8-20%. Using a previously established (1)H NMR structural-reporter-group concept, composite models, that include all identified structural features, were formulated for all mEPS molecules. Variations in the mEPS structures strongly affected the physical properties of the mEPSs. PMID- 19186952 TI - Vibrational energy dynamics of normal and deuterated liquid benzene. AB - Ultrafast Raman spectroscopy with infrared (IR) excitation is used to study vibrational energy dynamics of ambient temperature liquids benzene and benzene d(6). After IR pumping of a CH-stretch or CD-stretch parent excitation, the redistribution of vibrational energy is probed with anti-Stokes Raman. Ten benzene or 12 benzene-d(6) vibrations out of 30 total have large enough cross sections to be observed. The pathways, quantum yields, and lifetimes for energy transfer among these vibrations are quantified. Using a CCl(4) molecular thermometer, we demonstrate an ultrafast Raman calorimetry method which allows measurement of the rate that benzene vibrational energy is dissipated into the bath. On the basis of energy conservation, we then determine the time-dependent dissipation of aggregate vibrational energy from the unobserved, "invisible" vibrations. During the approximately 1 ps IR excitation process, vibrational energy is coherently redistributed to several vibrational modes ("coherently" means the rate is faster than (T(2))(-1) of the pumped transition). This energy is then further redistributed in an incoherent intramolecular vibrational relaxation process with a 6 ps T(1) time constant. The subsequent dynamics involve energy transfer processes accompanied by vibrational energy dissipation to the bath. This vibrational cooling process has a half-life of 30 ps in benzene and 20 ps in benzene-d(6), and thermalization is complete in approximately 100 ps. The observed strongly Raman-active vibrations have about the same amount of energy per mode as the invisible vibrations. The invisible vibrational energy in benzene decays somewhat faster than the observed energy. These two decay rates are about the same in benzene-d(6). PMID- 19186954 TI - Effects of ion exchange on the free volume and oxygen permeation in Nafion for fuel cells. AB - Variations of the free volume, O2 permeability, and structure of the Nafion membrane upon ion exchange of H+ with Na+ and K+ were studied. The free volume was quantified using the positron annihilation lifetime (PAL) technique, whereas the polymer structure was characterized by dynamic mechanical analysis (DMA), nanoindentation, and wide-angle X-ray diffraction (WAXD). It was found that the ion exchange significantly expands the free volume and at the same time decreases the O2 permeability. This is opposed to the simple free volume model in which the structure with less open volume is more amenable to lower permeability. Comparison of experimental data collected by different techniques revealed that not only the free volume but also the polymer stiffness plays an essential role in O2 permeation. PMID- 19186953 TI - Infrared spectra, integrated band intensities, and anharmonic force field of H2C=CHF. AB - The gas-phase infrared spectra of vinyl fluoride, H(2)C=CHF, have been examined at medium resolution in the range 400-8000 cm(-1). The assignment of the absorptions in terms of fundamental, overtone, and combination bands, assisted by quantum chemical calculations, is consistent all over the region investigated. Spectroscopic parameters, obtained from the analysis of partially resolved rotational structure of some bands, have been derived and compared with the corresponding calculated values. Accurate values of integrated band intensities have also been determined for the first time. High-level ab initio calculations with large basis sets have been performed. Correlated harmonic force fields have been obtained from coupled cluster CCSD(T) calculations with the cc-pVQZ basis set, while anharmonic force constants have been computed employing the less resource demanding cc-pVTZ basis set. A good agreement between the computed and the experimental data has been obtained including those for the integrated infrared band intensities. PMID- 19186955 TI - Supramolecular lone pair-pi/pi-pi/pi-anion assembly in a Mg(II)-malonate-2 aminopyridine-nitrate ternary system. AB - The solid-state structure of an alkaline-earth metal complex reveals the formation of a remarkable supramolecular framework based on concurrent lone pair pi, pi-pi, and pi-anion interactions whose stability has been investigated by density functional theory. PMID- 19186956 TI - Re: primary health care assessment and intervention in the dental office. Lambster IB, Wolf DL. (J Periodontol 2008;79:1825-1832). Primary care provided by periodontists/oral physicians. PMID- 19186957 TI - The role of bone decortication in enhancing the results of guided bone regeneration: a literature review. AB - BACKGROUND: Bone decortication is often performed as part of a guided bone regeneration (GBR) procedure. The biologic rationale for decortication of bone is to allow progenitor cells easy access to a GBR-treated site and to facilitate prompt angiogenesis. It also may enhance the physical connection between a bone graft and a recipient site. However, the concept of decortication prior to a GBR procedure is controversial because there are no human clinical trials to support its effectiveness, and there are opposing points of view derived from animal studies regarding its usefulness. METHODS: The literature was assessed to determine whether there are enough data to validate the rationale for using decortication of bone as an integral part of GBR procedures. Eight searches were performed seeking controlled clinical trials that addressed the ability of decortication to enhance GBR. RESULTS: Three controlled animal clinical trials were found that supported the use of decortication prior to performing GBR. Two controlled animal clinical trials were located that indicated decortication did not improve GBR procedures. No human controlled clinical trial was identified that addressed the ability of decortication to alter GBR procedures. The literature addressing the capacity of decortication to affect onlay grafting or wound healing also provided mixed results. CONCLUSION: There is conflicting information and not enough clinical trials to make a definitive determination as to the merits of bone decortication prior to GBR procedures. PMID- 19186958 TI - Results from the Periodontitis and Vascular Events (PAVE) Study: a pilot multicentered, randomized, controlled trial to study effects of periodontal therapy in a secondary prevention model of cardiovascular disease. AB - BACKGROUND: In the Periodontitis and Vascular Events (PAVE) pilot study, periodontal therapy was provided as an intervention in a secondary cardiac event prevention model through five coordinated cardiac-dental centers. METHODS: Subjects were randomized to either community care or protocol provided scaling and root planing to evaluate effects on periodontal status and systemic levels of high-sensitivity C-reactive protein (hs-CRP). RESULTS: After 6 months, there was a significant reduction in mean probing depth and extent of 4- or 5-mm pockets. However, there were no significant differences in attachment levels, bleeding upon probing, or extent of subgingival calculus comparing subjects assigned to protocol therapy (n = 151) to those assigned to community care (n = 152). Using intent-to-treat analyses, there was no significant effect on serum hs-CRP levels at 6 months. However, 48% of the subjects randomized to community care received preventive or periodontal treatments. Secondary analyses demonstrated that consideration of any preventive or periodontal care (i.e., any treatment) compared to no treatment showed a significant reduction in the percentage of people with elevated hs-CRP (values >3 mg/l) at 6 months. However, obesity nullified the periodontal treatment effects on hs-CRP reduction. The adjusted odds ratio for hs-CRP levels >3 mg/l at 6 months for any treatment versus no treatment among non-obese individuals was 0.26 (95% confidence interval: 0.09 to 0.72), adjusting for smoking, marital status, and gender. CONCLUSION: This pilot study demonstrated the critical role of considering obesity as well as rigorous preventive and periodontal care in trials designed to reduce cardiovascular risk. PMID- 19186959 TI - Periodontitis severity plus risk as a tooth loss predictor. AB - BACKGROUND: Tooth loss can be a consequence of the natural history of periodontitis. Stratification of periodontitis severity, risk, and tooth loss exists within the United States adult population, and tooth loss correlates to severity and risk. We evaluated the loss of teeth for a periodontitis-affected population categorized by the combination of severity and risk in which the subjects predominantly did not receive periodontal treatment. METHODS: The clinical records of 523 subjects enrolled in the Veterans Affairs Dental Longitudinal Study, covering a period of 15 years, were used. Disease severity, risk level, and the number of teeth lost for each subject were determined. RESULTS: A stepwise regression analysis showed that disease and risk scores predicted mean tooth loss rate. The P value for the disease score was <0.0005, and the P value for the risk score was 0.001. The ordinal logistic regression model showed that disease (P = 0.002) and risk scores (P = 0.000) were significantly associated with the probability of subjects losing a specific number of teeth. CONCLUSIONS: Tooth loss is more precisely and accurately predicted by the combination of risk score and periodontal disease score than by either score alone. The combined scores may be a surrogate variable for periodontal status. Because the scores are derived from routine clinical measurements, they may be useful for population surveillance and dynamics, practice management, patient care decisions, practice-based research, and the determination of treatment effectiveness and the factors required for successful treatment, resulting in improved oral health and higher clinician productivity and income. PMID- 19186960 TI - Magnesium-enriched hydroxyapatite compared to calcium sulfate in the healing of human extraction sockets: radiographic and histomorphometric evaluation at 3 months. AB - BACKGROUND: Reduction of alveolar height and width after tooth extraction may present problems for implant placement, especially in the anterior maxilla where bone volume is important for biologic and esthetic reasons. Different graft materials have been proposed to minimize the reduction in ridge volume. The aim of this study was to compare radiographic and histomorphometric results of magnesium-enriched hydroxyapatite (MHA) and calcium sulfate (CS) grafts in fresh sockets after tooth extractions. METHODS: Forty-five fresh extraction sockets with three bone walls were selected in 15 patients. A split-mouth design was used: 15 sockets on the right side of the jaw received MHA, 15 sockets on the left side received CS, and 15 random unfilled sockets were considered the control (C) group. Intraoral digital radiographs were taken at baseline and at 3 months after graft material placement. At 3 months, cylinder bone samples were obtained for histology and histomorphometry analysis. RESULTS: The difference in mean radiographic vertical bone level from baseline to 3 months was -2.48 +/- 0.65 mm in the CS group, -0.48 +/- 0.21 mm in the MHA group, and -3.75 +/- 0.63 mm in the unfilled C group. Statistically significant differences (P <0.05) were found between CS and MHA groups and between MHA and C groups. Histologic examination revealed bone formation in all treated sites; trabecular bone assessment did not differ among apical, mesial, and coronal portions of the specimens. Mean vital bone measurements for CS, MHA, and C groups were 45.0% +/- 6.5%, 40.0% +/- 2.7%, and 32.8% +/- 5.8%, respectively. Statistically significant differences (P <0.05) were found among all groups. Connective tissue percentages averaged 41.5% +/- 6.7% for the CS group, 41.3% +/- 1.3% for the MHA group, and 64.6% +/- 6.8% for the C group. Statistically significant differences (P <0.05) were found between CS and C groups and between MHA and C groups. The CS-grafted sockets showed 13.9% +/- 3.4% residual implant material, whereas the MHA-treated sockets showed 20.2% +/- 3.2% residual material. The difference between the groups was statistically significant (P <0.05). CONCLUSIONS: Radiographs revealed a greater reduction of alveolar ridge in the CS group than in the MHA group. Histologic examination showed more bone formation and faster resorption in the CS group and more residual implant material in the MHA group. PMID- 19186961 TI - Enamel matrix derivative versus bioactive ceramic filler in the treatment of intrabony defects: 12-month results. AB - BACKGROUND: This study examined the clinical efficacy of enamel matrix derivative (EMD) and bioactive ceramic filler (BCF) in the treatment of intrabony periodontal defects and evaluated factors influencing the treatment outcome. METHODS: Thirteen chronic periodontitis patients, 41 to 74 years of age, who had two proximal intrabony defects in different jaw quadrants with > or =3 mm vertical radiographic bone loss were selected for this study. After initial therapy, the sites in each patient were randomly assigned to EMD or BCF treatment. Clinical attachment level (CAL), probing depth (PD), tooth mobility (TM), gingival recession (GR), bleeding on probing, and dental plaque were recorded at baseline and at 6 and 12 months. At surgery, the intrabony component was characterized by recording the number of bony walls, distance in millimeters from the buccal crest (BC) to the most apical point of the defect, distance from the cemento-enamel junction (CEJ) to the BC, and the mesio-distal width of the defect at the level of the bony crest. RESULTS: BCF treatment resulted in a significant gain in proximal CAL (P = 0.005) and a reduction in proximal PD at 6 months (P <0.001), but there was no further improvement from 6 to 12 months. Paired comparisons by time for the EMD group revealed a significant reduction in proximal PD at 12 months (P = 0.001), whereas the gain in proximal CAL approached significance (P = 0.056). Mean GR increased significantly from baseline to 6 months in both groups (P = 0.001). Regression analysis revealed that within the EMD group, smoking and TM negatively influenced the gain of attachment, whereas within the BCF group, gingival recession increased with age, increasing CEJ to BC distance, and increasing mesial-distal width of the defect. CONCLUSIONS: The gain in proximal attachment after treating intrabony defects by flap surgery with BCF was significant (P = 0.004) and twice that following treatment with EMD (P = 0.056). Patient and site variables affected the clinical outcome differently. PMID- 19186962 TI - Stability of treated angular and horizontal bony defects: a retrospective radiographic evaluation in a private periodontal practice. AB - BACKGROUND: This retrospective radiographic evaluation provided a comparative assessment of long-term alterations of alveolar bone levels at sites with angular and horizontal bony defects that underwent open-flap debridement and did not undergo resective or regenerative periodontal therapy. METHODS: The investigation included 50 patients (27 met inclusion criteria) who had periodontal surgery and periodontal maintenance every 3 to 6 months and who presented with sites demonstrating radiographic angular defects and sites on adjacent or contralateral teeth that demonstrated horizontal bone loss. Subsequent to surgical therapy, alterations in bone levels at the alveolar crest of horizontal defects and at the base of angular defects were evaluated by comparing radiographs from the initial examination to those obtained after a period of periodontal maintenance (6 to 16 years; mean: 8.83 years). RESULTS: After periodontal therapy, sites with angular and horizontal alveolar bone loss had additional bone loss of 5.56% and 3.88%, respectively. CONCLUSION: Based on a defining criterion of 10%, the angular and horizontal sites were clinically equivalent with respect to the magnitude of osseous resorption that occurred after periodontal surgery and maintenance. PMID- 19186963 TI - Effect of anti-infective mechanical therapy on clinical parameters and cytokine levels in human peri-implant diseases. AB - BACKGROUND: The objectives of this study were to clinically and immunologically assess the effects of mechanical anti-infective therapies for mucositis and peri implantitis and to compare the levels of cytokines in untreated and treated peri implant diseased sites to healthy ones. METHODS: Titanium dental implants were assigned to one of the following groups: healthy (n = 10) = control; mucositis (n = 10) = mechanical debridement using abrasive sodium carbonate air-powder and resin curets; and peri-implantitis (n = 20) = open surgical debridement using abrasive sodium carbonate air-powder and resin curets. Visible plaque accumulation, marginal bleeding, bleeding on probing, suppuration, and probing depth were assessed at baseline for all groups and at 3 months after therapies for diseased groups. At these times, the total amounts of interleukin (IL)-4, 10, and -12, tumor necrosis factor-alpha (TNF-alpha), receptor activator of nuclear factor-kappa B ligand (RANKL), and osteoprotegerin (OPG) in the peri implant crevicular fluid (PICF) were measured by enzyme-linked immunosorbent assay. RESULTS: At 3 months, the anti-infective treatments resulted in a significant improvement in all clinical parameters for mucositis and peri implantitis (P <0.05). Moreover, the total amounts of TNF-alpha in PICF were significantly higher in untreated diseased implants compared to healthy ones, and the OPG/RANKL ratio was higher for healthy implants than for untreated peri implantitis (P <0.05). TNF-alpha levels were significantly reduced for both diseased groups (P <0.05), achieving the same level as the healthy group at 3 months after therapies (P >0.05). CONCLUSION: The proposed anti-infective therapies may locally modulate the levels of TNF-alpha and the OPG/RANKL ratio and improve clinical parameters around peri-implant tissues. PMID- 19186964 TI - Clinical evaluation of a modified coronally advanced flap alone or in combination with a platelet-rich fibrin membrane for the treatment of adjacent multiple gingival recessions: a 6-month study. AB - BACKGROUND: The aim of this study was to determine whether the addition of an autologous platelet-rich fibrin clot (PRF) to a modified coronally advanced flap (MCAF) (test group) would improve the clinical outcome compared to an MCAF alone (control group) for the treatment of multiple gingival recessions. METHODS: Twenty subjects, presenting three adjacent Miller Class I or II multiple gingival recessions of similar extent on both sides of the mouth, were enrolled in the study. The mean recession value at baseline was 2.9 +/- 1.1 mm for test sites and 2.5 +/- 0.9 mm for control sites. Each patient was treated on both sides by an MCAF technique; the combination treatment (with a PRF membrane) was applied on the test side. Probing depth (PD), recession width, clinical attachment level (CAL), keratinized gingival width, and gingival/mucosal thickness (GTH) were measured at baseline and at 6 months post-surgery. Gingival recession was measured at baseline and at 1, 3, and 6 months post-surgery. RESULTS: Mean root coverage after 1, 3, and 6 months was 81.0% +/- 16.6%, 76.1% +/- 17.7%, and 80.7% +/- 14.7%, respectively, at the test sites and 86.7% +/- 16.6%, 88.2% +/- 16.9%, and 91.5% +/- 11.4%, respectively, at the control sites. Differences between the two groups were statistically significant at 3 and 6 months. At 6 months, complete root coverage was obtained at 74.6% of the sites treated with the control procedure but at only 52.2% of the experimental sites. At 6 months, the increase in GTH was statistically significant when comparing the test sites (from 1.1 +/- 0.3 mm at baseline to 1.4 +/- 0.5 mm at 6 months) to the control sites (from 1.1 +/- 0.3 mm at baseline to 1.1 +/- 0.3 mm at 6 months). In the case of PD, there was no significant difference between the two groups at 6 months, but a significant CAL gain in favor of the control group was observed at that time. CONCLUSIONS: MCAF is a predictable treatment for multiple adjacent Miller Class I or II recession-type defects. The addition of a PRF membrane positioned under the MCAF provided inferior root coverage but an additional gain in GTH at 6 months compared to conventional therapy. PMID- 19186965 TI - Acellular dermal matrix graft for gingival augmentation: a preliminary clinical, histologic, and ultrastructural evaluation. AB - BACKGROUND: The aim of this study was to evaluate clinically, histologically, and ultrastructurally the integration process of the acellular dermal matrix used to increase the band of keratinized tissue while achieving gingival inflammation control. METHODS: Ten patients exhibiting a mucogingival problem with bands of keratinized tissue 5 mm. CONCLUSIONS: Stress and depression may be associated with periodontal destruction through behavioral and physiologic mechanisms. Addressing psychologic factors, such as depression, may be an important part of periodontal preventive maintenance. PMID- 19186967 TI - Periodontal status in subjects with hyperlipidemia. AB - BACKGROUND: Hyperlipidemia is a major risk factor for cardiovascular disease. Recent reports showed a possible association between periodontal disease and an increased risk for cardiovascular disease. The aim of this study was to evaluate whether hyperlipidemia has any influence on periodontal status. METHODS: Fifty one subjects with hyperlipidemia and 47 normolipidemic subjects participated in this study. Biochemical parameters, including plasma triglyceride, total cholesterol, low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) levels, and periodontal parameters, including plaque index (PI), probing depth (PD), clinical attachment level (CAL), and percentage of sites with bleeding on probing (BOP [%]), were evaluated. RESULTS: The mean values of PI, PD, CAL, and BOP (%) for the hyperlipidemic group were significantly higher than those for the control group. Plasma triglyceride, total cholesterol, and LDL-C levels were significantly and positively associated with PI, PD, BOP (%), and CAL. HDL-C was significantly, but negatively, associated with CAL. Plasma triglyceride level was significantly associated with PD and BOP (%) after further analyses. CONCLUSIONS: The results of our study showed that patients with mild or moderate hyperlipidemia manifested higher values of periodontal parameters compared to normolipidemic individuals. Further studies are needed to determine the effect of hyperlipidemia on periodontal disease. PMID- 19186968 TI - Proliferative verrucous leukoplakia: a potentially malignant disorder involving periodontal sites. AB - BACKGROUND: Proliferative verrucous leukoplakia (PVL) is a rare oral mucosa disorder, frequently involving periodontal sites, with a high rate of progression to oral squamous cell carcinoma (SCC) and verrucous carcinoma (VC). This article describes the clinical features and follow-up of a group of patients with PVL, with attention to the involvement of the mucosa covering the alveolar crest and its malignant transformation. METHODS: Patients were retrospectively evaluated for demographic data, risk habits (smoking and drinking), locations of PVL lesions, incidence, and locations of malignant transformation. Patients with malignant transformation were compared to a control group (patients affected by oral carcinoma without PVL). RESULTS: Forty-seven patients were enrolled; PVL lesions were most frequently observed on the alveolar crest (41/47 [87.2%]), with gingival involvement in 22 of 47 (46.8%) cases. Nineteen patients (40.4%) developed 41 malignant lesions; the alveolar crest was the most affected site (12/41 [29.3%]). Compared to controls, patients with PVL were more likely to develop VC (odds ratio [OR] = 6.61; 95% confidence interval [CI]: 1.23 to 65.52) than SCC (OR = 0.15; 95% CI: 0.02 to 0.82), and they showed a higher incidence of cancer on masticatory mucosa (OR = 6.49; 95% CI: 1.78 to 29.12), particularly gingiva (P = 0.007) and the hard palate (P = 0.017). CONCLUSIONS: The importance of PVL awareness for periodontists is underscored by the frequency of gingival involvement and the high prevalence of malignant transformation on masticatory mucosa, which usually can be suspected because of the onset of warning signs, such as rapid growth of verrucosity, area of erosion or ulceration, acquisition of red areas, induration, and positive response to toluidine blue staining. PMID- 19186969 TI - Aggressive periodontitis and chronic arthritis: blood mononuclear cell gene expression and plasma protein levels of cytokines and cytokine inhibitors. AB - BACKGROUND: Cytokines and cytokine inhibitors have been associated with many immunoinflammatory diseases. In the present study, we examined whether peripheral blood mononuclear cell (PBMC) gene expression mirrors the corresponding plasma levels of clinically important pro- and anti-inflammatory cytokines and cytokine receptors in patients with periodontitis and patients with arthritis representing two examples of chronic inflammatory diseases, such as periodontitis and arthritis. To identify possible disease-specific characteristics of subjects with periodontitis relative to subjects with chronic inflammation in general, patients with arthritis (juvenile idiopathic arthritis [JIA] and rheumatoid arthritis [RA]) were included. METHODS: The study population consisted of white adults <35 years of age diagnosed with localized aggressive periodontitis (LAgP; n = 18), generalized aggressive periodontitis (GAgP; n = 27), JIA (n = 10), and RA (n = 23) and healthy controls (n = 25). PBMC transcripts of interleukin (IL) 1 alpha (IL1A), IL 1 beta (IL1B), IL 1 receptor antagonist (IL1RN), IL6, IL10, tumor necrosis factor alpha (TNFA), TNF alpha receptor I (TNFRI), and TNFRII were measured using real-time reverse transcription-polymerase chain reaction and compared to the corresponding plasma protein levels measured by enzyme-linked immunosorbent assay and a multiplex antibody bead assay. RESULTS: Compared to controls, soluble (s) TNF-RII levels were significantly elevated in patients with GAgP (P = 0.001) or JIA (P = 0.002), and PBMC TNFA transcript levels were lower in patients with JIA (P = 0.001). A negative correlation was found between IL6 expression and IL-6 plasma levels in patients with JIA versus controls, and a positive correlation/association was found between TNFRI expression and sTNF-RI plasma levels in patients with LAgP and RA. CONCLUSIONS: The study demonstrated only a few changes in the PBMC expression of various cytokine and cytokine inhibitor genes in aggressive periodontitis and chronic arthritis compared to controls. There were a few similarities among disease groups, and no direct and simple correlation between cellular transcripts and proteins was found. PMID- 19186970 TI - Levels of interferon-gamma and transcription factor T-bet in progressive periodontal lesions in patients with chronic periodontitis. AB - BACKGROUND: Periodontitis is an infection with an episodic pattern of tissue support destruction. During the generation of a primary CD4(+) T helper 1 (Th1) response, interferon-gamma (IFN-gamma) acts as a positive regulator by selectively inducing Th1 differentiation through increased transcription of T bet. The aims of this work were to determine IFN-gamma levels in samples of gingival crevicular fluid (GCF) and to determine IFN-gamma and transcription factor T-bet expression in gingival tissue from patients undergoing the progression of chronic periodontitis. METHODS: One hundred six patients with moderate or advanced chronic periodontitis were selected. Periodontitis was characterized by at least six sites with probing depth >or=5 mm, clinical attachment loss >or=3 mm, and radiographic bone loss. Periodontitis progression was determined by the tolerance method. GCF was collected using a paper strip, and enzyme-linked immunosorbent assay was performed to determine the total amount of IFN-gamma. Gingival biopsies were obtained from patients for real-time reverse transcription-polymerase chain reaction to determine IFN-gamma and T-bet expression. Statistical analysis was performed using statistical software. Data were expressed as subject means +/- SD. The chi(2) and Student t tests were used. RESULTS: The total amount and concentration of cytokine IFN-gamma were significantly higher in active sites than in inactive sites (99.90 versus 68.90 pg; P = 0.03; 106.62 pg/mg versus 75.64 pg/mg, P = 0.04, respectively). Active sites showed a significantly lower Delta cycle threshold (Ct) of IFN-gamma than inactive sites (P = 0.04), whereas the expression of transcription factor T-bet was increased 1.42-fold in active sites compared to inactive sites. CONCLUSION: The total amount and concentration of cytokine IFN-gamma in GCF samples and transcription factor T-bet expression were increased in progressive periodontal lesions in patients with chronic periodontitis. PMID- 19186971 TI - Changes in inflammatory mediators in peri-implant fluid after implant insertion. AB - BACKGROUND: Endosseous dental titanium implants have revolutionized restorative dentistry and have made a significant impact on improved patient care. The aim of this study was to compare and evaluate the influence of the placement technique on periodontal health. METHODS: A baseline examination was performed in patients with submerged and non-submerged titanium implants, including an evaluation of plaque index (PI), gingival index (GI), periodontal probing depth (PD), clinical attachment level (CAL), and bone level, as well as histamine and arachidonic acid metabolite concentrations, in the peri-implant crevicular fluid. Examinations were repeated after 12, 24, and 36 months. RESULTS: Bone loss was significantly higher in the submerged group relative to the non-submerged group at 3 years (P <0.01), with a slight increase at 24 months. All clinical parameters were significantly higher in the submerged group relative to the non-submerged group at 24 and 36 months (P <0.05 for PI; P <0.01 for GI, PD, and CAL). The mean levels of histamine and other inflammatory mediators were significantly higher, whereas 15(S)-hydroxy-5,8,11,13-eicosatetraenoic acid concentrations were significantly reduced in the submerged group, with a high correlation with periodontal indices at 24 and 36 months (P <0.001). CONCLUSION: This longitudinal study suggested that submerged implants present a number of risks for periodontal complications compared to non-submerged implants, which can be evidenced by inflammatory mediator variations in the peri-implant crevicular fluid. PMID- 19186972 TI - Levels of serum interleukin (IL)-6 and gingival crevicular fluid of IL-1beta and prostaglandin E(2) among non-smoking subjects with gingivitis and type 2 diabetes. AB - BACKGROUND: The goal of this study was to assess whether non-smoking patients with type 2 diabetes present with increased levels of local and systemic proinflammatory mediators and, if so, whether such an increase is associated with enhanced clinical gingival inflammation compared to non-smoking patients without diabetes. METHODS: We used a cross-sectional database consisting of 725 self reported lifelong non-smokers aged 53 to 74 years. Gingival crevicular fluid (GCF) levels of interleukin (IL)-1beta and prostaglandin E(2) (PGE(2)) and serum levels of IL-6 were measured using enzyme-linked immunosorbent assay. No participant had probing depth >3 mm. Participants with bleeding on probing (BOP) in <10% of sites were classified as healthy, whereas those with BOP in >or=10% of sites were defined as having biofilm-gingival interface (BGI) gingivitis. RESULTS: Approximately 53% (n = 385) and 11% (n = 80) of the sample had BGI gingivitis and type 2 diabetes, respectively. The mean age-adjusted level of GCF IL-1beta was significantly elevated in the diabetic group compared to the non diabetic group (P = 0.048), but serum IL-6 (P = 0.14) and GCF PGE(2) were not (P = 0.98). The mean GCF IL-1beta and PGE(2) levels were significantly elevated in subjects with BGI gingivitis (136.2 +/- 112.9 ng/ml and 277.2 +/- 187.2 ng/ml, respectively) compared to subjects with gingival health (95.9 +/- 82.9 ng/ml and 205.7 +/- 149.6 ng/ml, respectively), regardless of diabetic status (P <0.001 for both). However, serum IL-6 was elevated in subjects with BGI gingivitis compared to subjects with gingival health only among subjects with diabetes (2.9 +/- 3.2 pg/ml versus 1.5 +/- 1.4 pg/ml; P = 0.008). With the exception of serum IL-6 in subjects without diabetes, an increase in the levels of proinflammatory mediators was associated with increased odds of having BGI gingivitis. The associations were stronger in the diabetic group. CONCLUSIONS: Type 2 diabetes may increase the host inflammatory response to oral biofilm, which, in turn, may exacerbate preconditions associated with gingivitis in susceptible individuals. Furthermore, systemic inflammation, as demonstrated by the increased level of serum IL-6, is associated with BGI gingivitis among non-smoking patients with diabetes. PMID- 19186974 TI - Placement of two-part implants in sites with different buccal and lingual bone heights. AB - BACKGROUND: Recipient sites for implants often exhibit a vertical discrepancy between the buccal and lingual bone crest. The potential to preserve the lingual bone when placing the implant within the dimension determined by the buccal bone in such sites remains to be evaluated. METHODS: Five dogs were included in this study. The premolars were extracted on one side of the mandible, and a buccal defect was prepared by the resection of a 2-mm-high portion of the buccal bone wall. Three months later, two test (surface modification on the shoulder part of the implant) and two control implants were placed within the area of the buccal bone defect. Following placement, the implant margin at the buccal side coincided with the buccal bone crest, whereas the implant margin at the lingual side was in a subcrestal position. Regular abutments with a turned surface were connected to the control implants, whereas experimental abutments with a modified surface were connected to the test implants. A plaque-control program was initiated. Four months later, biopsies were obtained and prepared for histologic analysis. RESULTS: The marginal bone level was located at a more coronal position at the lingual aspect than at the buccal aspect of the test implants, whereas the buccal and lingual bone levels were similar at control implants. CONCLUSION: It is suggested that different marginal bone levels are obtainable at the lingual and buccal aspects when two-part implants with suitable surface characteristics are placed in sites with different buccal and lingual bone heights. PMID- 19186973 TI - Effects of ascorbic acid on gingival melanin pigmentation in vitro and in vivo. AB - BACKGROUND: Gingival melanin pigmentation may cause esthetic concerns, even if no serious medical problem is present. As an inhibitor of melanin formation, ascorbic acid is often used to treat skin melanin pigmentation. Thus, the present study investigated the effects of ascorbic acid on gingival melanin pigmentation in vitro and in vivo. METHODS: The effects of ascorbic acid on melanin formation were evaluated in vitro in B16 mouse melanoma cells and three-dimensional human skin models. In addition, a clinical trial was performed to investigate the inhibitory effects of a gel containing ascorbic acid 2-glucoside (AS-G gel) on gingival melanin pigmentation. This study used a double-masked, split-mouth design on 73 subjects with symmetric gingival melanin pigmentation. AS-G gel was applied to one side of the gingiva for 12 weeks, whereas placebo gel was applied to the other side as a control. Luminance (L*)-value, which describes the lightness of gingiva, was determined by spectrophotometry to obtain an objective measure of melanin pigmentation every 4 weeks. RESULTS: Ascorbic acid significantly inhibited tyrosinase activity and melanin formation in B16 mouse melanoma cells (P <0.01 and P <0.05, respectively). The inhibitory effects of ascorbic acid on melanin formation were also significant in three-dimensional human skin models (P <0.01). Moreover, in the clinical trial, a significant relative change in pigmentation was seen after 4 weeks with the application of AS G gel compared to placebo (L*-value ratio). CONCLUSION: Ascorbic acid (AS-G) has potential for the treatment of gingival melanin pigmentation. PMID- 19186975 TI - Metronidazole-loaded bioabsorbable films as local antibacterial treatment of infected periodontal pockets. AB - BACKGROUND: Periodontal disease is infectious in nature and leads to an inflammatory response. It arises from the accumulation of subgingival bacterial plaque and leads to the loss of attachment, increased probing depth, and bone loss. It is one of the world's most prevalent chronic diseases. In this study we developed and studied metronidazole-loaded 50/50 poly(DL-lactide-co-glycolide) (PDLGA), 75/25 PDLGA, and poly(DL-lactic acid) (PDLLA) films. These films are designed to be inserted into the periodontal pocket and treat infections with controlled-release metronidazole for >or=1 month. METHODS: The structured films were prepared using the solution-casting technique. Concentrated solutions and high solvent-evaporation rates were used to get most of the drug located in the bulk, i.e., in whole film's volume. The effects of copolymer composition and drug content on the release profile, cell growth, and bacterial inhibition were investigated. RESULTS: The PDLLA and 75/25 PDLGA films generally exhibited a low- or medium-burst release followed by a moderate release at an approximately constant rate, whereas the 50/50 PDLGA films exhibited a biphasic release profile. The drug released from films loaded with 10% weight/weight metronidazole resulted in a significant decrease in bacterial viability within several days. When exposed to human gingival fibroblasts in cell culture conditions, these films maintained their normal fibroblastic features. CONCLUSIONS: This study enabled the understanding of metronidazole-release kinetics from bioabsorbable polymeric films. The developed systems demonstrated good biocompatibility and the ability to inhibit Bacteroides fragilis growth; therefore, they may be useful in the treatment of periodontal diseases. PMID- 19186976 TI - Identification of N-methyl-D-aspartate receptor subunit in human periodontal ligament fibroblasts: potential role in regulating differentiation. AB - BACKGROUND: Periodontal ligament fibroblasts (PDLFs), which can be differentiated into osteoblasts, are crucial cells for the regeneration of the periodontal tissue. Although N-methyl-D-aspartate (NMDA) receptors were reported to be involved in bone formation by affecting osteoblasts, the existence and function of NMDA receptors in PDLFs have not been confirmed. The purpose of this study was to examine the expression of NMDA receptors and their role in human PDLFs. METHODS: Human PDLFs were cultured and evaluated to identify the subunits of NMDA receptors (NR) by reverse transcription-polymerase chain reaction, Western blot analysis, and immunocytochemistry. Then, the cells were assigned to four different groups: a control media group, a control media with NMDA receptor antagonist group, a differentiation media group, and a differentiation media with NMDA receptor antagonist group. Cell proliferation assay, alkaline phosphatase (ALP) activity analysis, and mineralization assay were performed to determine whether NMDA receptors affected the function of PDLFs. RESULTS: NR1, NR2B, and NR2D were detected in human PDLFs. There was no statistically significant difference in proliferation among the groups. However, the NMDA receptor antagonist-treated group showed a significant reduction in ALP activity (P <0.05). Moreover, the NMDA receptor antagonist-supplemented group presented no mineralization. CONCLUSIONS: This study revealed the existence of NMDA receptors in human PDLFs and specified their subunits. Moreover, NMDA receptors had a significant influence on the differentiation and mineralization of human PDLFs but did not affect their proliferation. These results suggest that NMDA receptors may play an important role in the differentiation and mineral tissue formation of human PDLFs. PMID- 19186977 TI - Minimally invasive flapless implant placement: follow-up results from a multicenter study. AB - BACKGROUND: The placement of implants using a minimally invasive flapless approach has the potential to reduce operative bleeding and postoperative discomfort and minimize crestal bone loss. This article presents follow-up data on a prospective clinical study of implants placed using a flapless procedure. METHODS: The original study reported on 57 patients (33 female patients with an age range of 24 to 86 years; 24 male patients with an age range of 27 to 81 years) recruited from three clinical centers (Tucson, Arizona; Gothenburg, Sweden; and Tel Aviv, Israel) who received 79 implants. After an average of 3 years and 8 months, the patients were contacted and invited to return to their respective clinics for reexamination. Thirty-seven patients with 52 implants returned for a follow-up examination; the remaining 20 patients (27 implants) were not available for reexamination and were considered study drop-outs. RESULTS: The cumulative survival rate at the 3- to 4-year follow-up examination remains at 98.7%, reflecting the loss of one implant. The mean probing depth at abutment connection was 2.2 mm, as reported in the initial study (examination 2 at approximately 2 years postplacement); it was 2.4 mm at the 3- to 4-year second follow-up examination. This change was not clinically or statistically significant. Bleeding score changes also were not significant between the two intervals. The average crestal bone level was -0.7 mm at examination 2 and -0.8 mm at examination 3, a change that approached significance (P <0.06). CONCLUSIONS: Minimally invasive flapless surgery offers patients the possibility of high implant predictability with clinically insignificant crestal bone loss for up to 4 years. Proper diagnosis and treatment planning are key factors in achieving predictable outcomes. PMID- 19186978 TI - Single-flap approach with buccal access in periodontal reconstructive procedures. AB - BACKGROUND: New surgical techniques have been developed to optimize primary closure as well as to minimize the surgical trauma in the reconstructive procedures of periodontal intraosseous defects. Recently, we proposed a minimally invasive procedure, the single-flap approach (SFA), specifically indicated when the defect extension is prevalent on the buccal or oral side. The basic principle of the SFA is the elevation of a flap to access the defect only on one side (buccal or oral), leaving the opposite side intact. The present case series reports preliminary data on the clinical effectiveness of SFA with buccal access in conjunction with a collagen membrane and a hydroxyapatite (HA)-based biomaterial in the reconstructive treatment of deep periodontal intraosseous defects. METHODS: Ten intraosseous defects in 10 patients were accessed with a buccal SFA and treated with a collagen membrane and an HA-based graft biomaterial. The follow-up period following the regenerative procedure ranged from 6 to 14 months (mean: 10.0 +/- 3.0 months). RESULTS: Clinical attachment level (CAL) decreased from 11.2 +/- 2.6 mm presurgery to 6.4 +/- 1.9 mm post surgery. Probing depth was 9.0 +/- 2.8 mm before surgery and 3.8 +/- 1.5 mm post surgery. Gingival recession increased from 2.2 +/- 1.9 mm presurgery to 2.6 +/- 1.3 mm post-surgery. CONCLUSIONS: Challenging intraosseous defects, surgically accessed with a buccal SFA and treated with a combined graft/guided tissue regeneration technique, may heal with a substantial CAL gain. Limited postsurgical recession indicates that SFA may represent a suitable option to surgically treat defects in areas with high esthetic demands. PMID- 19186979 TI - Identification of abundant alkyl ether glycerophospholipids in the human lens by tandem mass spectrometry techniques. AB - Previous studies have shown that the human lens contains glycerophospholipids with ether linkages. These lipids differ from conventional glycerophospholipids in that the sn-1 substituent is attached to the glycerol backbone via an 1-O alkyl or an 1-O-alk-1'-enyl ether rather than an ester bond. The present investigation employed a combination of collision-induced dissociation (CID) and ozone-induced dissociation (OzID) to unambiguously distinguish such 1-O-alkyl and 1-O-alk-1'-enyl ethers. Using these methodologies the human lens was found to contain several abundant 1-O-alkyl glycerophosphoethanolamines, including GPEtn(16:0e/9Z-18:1), GPEtn(11Z-18:1e/9Z-18:1), and GPEtn(18:0e/9Z-18:1), as well as a related series of unusual 1-O-alkyl glycerophosphoserines, including GPSer(16:0e/9Z-18:1), GPSer(11Z-18:1e/9Z-18:1), GPSer(18:0e/9Z-18:1) that to our knowledge have not previously been observed in human tissue. Isomeric 1-O-alk-1' enyl ethers were absent or in low abundance. Examination of the double bond position within the phospholipids using OzID revealed that several positional isomers were present, including sites of unsaturation at the n-9, n-7, and even n 5 positions. Tandem CID/OzID experiments revealed a preference for double bonds in the n-7 position of 1-O-ether linked chains, while n-9 double bonds predominated in the ester-linked fatty acids [e.g., GPEtn(11Z-18:1e/9Z-18:1) and GPSer(11Z-18:1e/9Z-18:1)]. Different combinations of these double bond positional isomers within chains at the sn-1 and sn-2 positions point to a remarkable molecular diversity of ether-lipids within the human lens. PMID- 19186980 TI - Integrated electrokinetic sample focusing and surface plasmon resonance imaging system for measuring biomolecular interactions. AB - Label-free biomolecular binding measurement methods, such as surface plasmon resonance (SPR), are becoming increasingly more important for the estimation of real-time binding kinetics. Recent advances in surface plasmon resonance imaging (iSPR) are emerging for label-free microarray-based assay applications, where multiple biomolecular interactions can be measured simultaneously. However, conventional iSPR microarray systems rely on protein printing techniques for ligand immobilization to the gold imaging surface and external pumps for analyte transport. In this article, we present an integrated microfluidics and iSPR platform that uses only electrokinetic transport and guiding of ligands and analytes and, therefore, requires only electrical inputs for sample transport. An important advantage of this new approach, compared to conventional systems, is the ability to direct a single analyte to a specific ligand location in the microarray, which can facilitate analysis parallelization. Additionally, this simple approach does not require complicated microfluidic channel arrangements, external pumps, or valves. As a demonstration, kinetics and affinity have been extracted from measured binding responses of human IgG and goat antihuman IgG using a simple 1:1 model and compared to responses measured with conventional pressure driven analyte transport. The measured results indicate similar binding kinetics and affinity between the electrokinetic and pressure-driven sample manipulation methods and no cross contamination to adjacent measurement locations has been observed. PMID- 19186981 TI - Shear stress, reactive oxygen species, and arterial structure and function. AB - Shear stress is well known to be a key factor in the regulation of small-artery tone and structure. Although nitric oxide is a major endothelium-derived factor involved in short- and long-term regulation of vascular caliber, it is clear that other mechanisms also can be involved. This review discusses the evidence for endothelium-derived reactive oxygen species (ROS) as mediators for shear dependent arterial tone and remodeling. The work focuses on resistance vessels, because their caliber determines local perfusion. However, work on large vessels is included where needed. Attention is given to the shear-stress levels and profiles that exist in the arterial system and the differential effects of steady and oscillating shear on NO and ROS production. We furthermore address the relation between microvascular tone and remodeling and the effect of ROS and inflammation on the activity of remodeling enzymes such as matrix metalloproteinases and transglutaminases. We conclude that future work should address the role of H(2)O(2) as an endothelium-derived factor mediating tone and influencing structure of small arteries over the long term. PMID- 19186982 TI - Reprogramming of human somatic cells using human and animal oocytes. AB - There is renewed interest in using animal oocytes to reprogram human somatic cells. Here we compare the reprogramming of human somatic nuclei using oocytes obtained from animal and human sources. Comparative analysis of gene expression in morula-stage embryos was carried out using single-embryo transcriptome amplification and global gene expression analyses. Genomic DNA fingerprinting and PCR analysis confirmed that the nuclear genome of the cloned embryos originated from the donor somatic cell. Although the human-human, human-bovine, and human rabbit clones appeared morphologically similar and continued development to the morula stage at approximately the same rate (39, 36, and 36%, respectively), the pattern of reprogramming of the donor genome was dramatically different. In contrast to the interspecies clones, gene expression profiles of the human-human embryos showed that there was extensive reprogramming of the donor nuclei through extensive upregulation, and that the expression pattern was similar in key upregulation in normal control embryos. To account for maternal gene expression, enucleated oocyte transcriptome profiles were subtracted from the corresponding morula-stage embryo profiles. t-Test comparisons (median-normalized data @ fc>4; p<0.005) between human in vitro fertilization (IVF) embryos and human-bovine or human-rabbit interspecies somatic cell transfer (iSCNT) embryos found between 2400 and 2950 genes that were differentially expressed, the majority (60-70%) of which were downregulated, whereas the same comparison between the bovine and rabbit oocyte profiles found no differences at all. In contrast to the iSCNT embryos, expression profiles of human-human clones compared to the age-matched IVF embryos showed that nearly all of the differentially expressed genes were upregulated in the clones. Importantly, the human oocytes significantly upregulated Oct-4, Sox-2, and nanog (22-fold, 6-fold, and 12-fold, respectively), whereas the bovine and rabbit oocytes either showed no difference or a downregulation of these critical pluripotency-associated genes, effectively silencing them. Without appropriate reprogramming, these data call into question the potential use of these discordant animal oocyte sources to generate patient specific stem cells. PMID- 19186983 TI - Site-specific characterization of HIV-1 nucleocapsid protein binding to oligonucleotides with two binding sites. AB - The nucleocapsid protein (NC) of HIV-1 is a highly conserved protein essential for the virus life cycle that constitutes an attractive target for new antiviral agents. Most NC functions rely on its binding to the HIV-1 genomic RNA and its DNA copies that contain multiple and possibly interdependent binding sites. Therefore, a detailed understanding of NC binding requires a site-specific experimental approach. We have recently shown that 2-aminopurine (2Ap), a fluorescent adenine analogue, can site-selectively probe the binding of NC. Here, we introduced 2Ap at various positions of model single-stranded dodecanucleotides containing two TG motifs which constitute putative specific binding sites. Steady state and time-resolved fluorescence experiments indicated that NC binding strongly increased the fluorescence quantum yield of 2AP by reducing the dynamic quenching of 2Ap by its close neighbors and slowing the picosecond to nanosecond conformational fluctuations of the oligonucleotides. The dodecanucleotides were found to bind two NC molecules at physiological salt concentrations, confirming the preferential binding of NC to TG motifs and an occluded binding site size for NC of five to six bases. Using the NC-induced changes in 2Ap fluorescence, we determined the microscopic affinity constants of the individual binding sites and showed that affinities can significantly differ from one site to another within the same dodecanucleotide, depending on the position of the TG dinucleotide and the nature of its close neighbors. Moreover, our data suggest that binding of NC even to close binding sites shows no strong cooperativity. PMID- 19186984 TI - Enthalpy-driven RNA folding: single-molecule thermodynamics of tetraloop-receptor tertiary interaction. AB - RNA folding thermodynamics are crucial for structure prediction, which requires characterization of both enthalpic and entropic contributions of tertiary motifs to conformational stability. We explore the temperature dependence of RNA folding due to the ubiquitous GAAA tetraloop-receptor docking interaction, exploiting immobilized and freely diffusing single-molecule fluorescence resonance energy transfer (smFRET) methods. The equilibrium constant for intramolecular docking is obtained as a function of temperature (T = 21-47 degrees C), from which a van't Hoff analysis yields the enthalpy (DeltaH degrees) and entropy (DeltaS degrees) of docking. Tetraloop-receptor docking is significantly exothermic and entropically unfavorable in 1 mM MgCl(2) and 100 mM NaCl, with excellent agreement between immobilized (DeltaH degrees = -17.4 +/- 1.6 kcal/mol, and DeltaS degrees = -56.2 +/- 5.4 cal mol(-1) K(-1)) and freely diffusing (DeltaH degrees = -17.2 +/- 1.6 kcal/mol, and DeltaS degrees = -55.9 +/- 5.2 cal mol(-1) K(-1)) species. Kinetic heterogeneity in the tetraloop-receptor construct is unaffected over the temperature range investigated, indicating a large energy barrier for interconversion between the actively docking and nondocking subpopulations. Formation of the tetraloop-receptor interaction can account for approximately 60% of the DeltaH degrees and DeltaS degrees of P4-P6 domain folding in the Tetrahymena ribozyme, suggesting that it may act as a thermodynamic clamp for the domain. Comparison of the isolated tetraloop-receptor and other tertiary folding thermodynamics supports a theme that enthalpy- versus entropy-driven folding is determined by the number of hydrogen bonding and base stacking interactions. PMID- 19186987 TI - Chiral ligand exchange potentiometric aspartic acid sensors with polysiloxane films containing a chiral ligand N-carbobenzoxy-aspartic acid. AB - An enantioselective molecular sensor was fabricated by inserting a chiral ligand, N-carbobenzoxy-L-aspartic acid (N-CBZ-L-Asp) or N-CBZ-D-Asp, into an octadecylsiloxane (ODS) monolayer by polysiloxane film immobilization (PFI). The resulting system can recognize one enantiomer of aspartic acids (Asps) due to the chiral ligand exchange reaction at the N-CBZ-L-/D-Asp modified indium-tin oxide (ITO)-coated electrode. The enantioselective formation of diastereoisomeric complexes of Cu(II) with target enantiomers, in here L-/D-Asps, and N-CBZ-L-/D Asp immobilized by PFI on the ITO electrode. Those diastereoisomeric complexes have different thermodynamic stabilities and Nernst factors and thus enable the sensors to convert the enantioselective recognition event into potential changes by detecting Asp enantiomers in a concentration range of (4.0 x 10(-8))-(8.9 x 10(-5)) M without any pre- or postseparation process. The enantiomeric selectivity coefficients of the sensors for the counterisomers were in the range of (4.0 x 10(-5))-(5.0 x 10(-5)). PMID- 19186986 TI - Cyclic stretch, reactive oxygen species, and vascular remodeling. AB - Blood vessels respond to changes in mechanical load from circulating blood in the form of shear stress and mechanical strain as the result of heart propulsions by changes in intracellular signaling leading to changes in vascular tone, production of vasoactive molecules, and changes in vascular permeability, gene regulation, and vascular remodeling. In addition to hemodynamic forces, microvasculature in the lung is also exposed to stretch resulting from respiratory cycles during autonomous breathing or mechanical ventilation. Among various cell signaling pathways induced by mechanical forces and reported to date, a role of reactive oxygen species (ROS) produced by vascular cells receives increasing attention. ROS play an essential role in signal transduction and physiologic regulation of vascular function. However, in the settings of chronic hypertension, inflammation, or acute injury, ROS may trigger signaling events that further exacerbate smooth muscle hypercontractility and vascular remodeling associated with hypertension and endothelial barrier dysfunction associated with acute lung injury and pulmonary edema. These conditions are also characterized by altered patterns of mechanical stimulation experienced by vasculature. This review will discuss signaling pathways regulated by ROS and mechanical stretch in the pulmonary and systemic vasculature and will summarize functional interactions between cyclic stretch- and ROS-induced signaling in mechanochemical regulation of vascular structure and function. PMID- 19186989 TI - Structural basis of the broad specificity of a general odorant-binding protein from honeybee. AB - General odorant-binding proteins (GOBPs) are believed to transport a wide range of volatile hydrophobic molecules across the aqueous sensillum lymph toward olfactory receptors in insects. GOBPs are involved in the first step of odorant recognition, which has a great impact in agriculture and in insect-mediated human disease control. We report here the first structural study of a GOBP, the honeybee ASP2, in complex with a small hydrophilic ligand. The overall fold of the NMR structure of ASP2 consists of the packing of six alpha-helices creating an internal cavity and closely resembles that of the related pheromone-binding proteins (PBPs). The predominantly hydrophobic internal cavity of ASP2 provides additional possible interactions (pi-stacking, electrostatic contact) for ligand binding. We also show that the internal cavity of ASP2 has the ability to bind ligands of different structures and properties, including a hydrophobic component of the floral scent [2-isobutyl-3-methoxypyrazine (IBMP)] and a small hydrophilic ligand. We further demonstrate that IBMP binds ASP2 with two stable alternative conformations inside the ASP2 binding pocket. The (15)N NMR relaxation study suggests that significant backbone mobility occurs at the ligand entry site at the millisecond rate, which likely plays a role in the recognition and the uptake release mechanism of ligands by ASP2. We propose that the broad ligand specificity of GOBPs compared with PBPs is conferred by the cumulative effects of weak nonspecific protein-ligand interactions and of enhanced protein internal dynamics at the ligand entry site. PMID- 19186990 TI - Quantification of the post-translational addition of amino acids to proteins by MALDI-TOF mass spectrometry. AB - Aminoacyl-tRNA protein transferases catalyze the post-translational addition of amino acids to proteins. The eubacterial leucyl/phenylalanyl-tRNA-protein transferase (L/F transferase) catalyzes the transfer of leucine or phenylalanine from their respective aminoacylated tRNAs to the N-termini of substrate proteins possessing an N-terminal lysine or arginine amino acid. Conventional assays to quantify L/F transferase activity involve measuring radioactive amino acid incorporation into substrate proteins. We have developed a quantitative matrix assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry procedure to measure the enzymatic activity of L/F transferase. The procedure utilizes stable isotope labeled substrate and internal standard peptides. The method is used to determine the kinetic parameters of k(cat) and K(m) for the enzymatic transfer of phenylalanine and three unnatural amino acid derivatives from an aminoacyl-tRNA to a peptide substrate. PMID- 19186992 TI - New treatment of periodontal diseases by using NF-kappaB decoy oligodeoxynucleotides via prevention of bone resorption and promotion of wound healing. AB - Nuclear factor-kappa B (NF-kappaB) is involved in osteoclast differentiation and activation. Thus, the blockade of the NF-kappaB pathway might be a novel therapeutic strategy for treating bone metabolic diseases. Periodontitis is subgingival inflammation caused by bacterial infection; this disease also is thought to be a chronic focal point responsible for systemic diseases. In this study, NF-kappaB decoy oligodeoxynucleotides (ODNs) were topically applied for experimental periodontitis in a debris-accumulation model and wound healing in a bone-defect model of beagle dogs to investigate the effect of decoy ODN on bone metabolism. Application of NF-kappaB decoy ODN significantly reduced interleukin 6 activity in crevicular fluid and improved alveolar bone loss in the analysis of dental radiographs and DEXA. Direct measurement of exposed root that lost alveolar bone support revealed that NF-kappaB decoy treatment dramatically protected bone from loss. In a bone-defect model, NF-kappaB decoy ODN promoted the healing process as compared with control scrambled decoy in micro-CT analysis. Overall, inhibition of NF-kappaB by decoy strategy prevented the progression of bone loss in periodontitis and promoted the wound healing in bone defects through the inhibition of osteoclastic bone resorption. Targeting of NF kappaB might be a potential therapy in various bone metabolic diseases. PMID- 19186995 TI - The paracrine role of Tie-2-expressing monocytes in tumor angiogenesis. AB - Literature data have demonstrated that tumor neovascularization is regulated in part by myelomonocytic cells. Luigi Naldini's group has reported the identification in human peripheral blood of a novel subset of Tie-2-expressing monocytes (TEMs) that promote angiogenesis in paracrine manner. Although recruited to tumors in lower numbers than tumor-associated macrophages (TAMs), TEMs are a more potent source of proangiogenic signals, suggesting that they significantly contribute to tumor angiogenesis. Moreover, TEMs, while stimulating angiogenesis, do not actively incorporate into blood vessels and this subpopulation of Tie-2+ cells, rather than bone marrow-derived endothelial progenitor cells (EPCs), which are incorporated in new-forming blood vessels, promote tumor neovascularization through the release of proangiogenic factors. PMID- 19186993 TI - Thioredoxin suppresses the contact hypersensitivity response by inhibiting leukocyte recruitment during the elicitation phase. AB - Thioredoxin, a redox-regulating protein that scavenges reactive oxygen species, appears to show an excellent antiinflammatory effect in treating animal models of various human inflammatory diseases. The aim of this study was to clarify whether thioredoxin is useful for treating inflammatory skin diseases, such as contact dermatitis, caused by epicutaneous exposure to environmental and occupational antigens. The allergic contact hypersensitivity response was suppressed in thioredoxin-transgenic mice. This suppressive effect of thioredoxin appeared to be via the inhibition of the efferent limb of contact hypersensitivity because administration of recombinant thioredoxin suppressed the inflammatory response in the elicitation phase but not in the induction phase. Adoptive-transfer studies revealed that the host environment, but not donor leukocytes, is critical in this suppressive effect. In thioredoxin-transgenic mice, the infiltration of neutrophils in the elicitation site was diminished, whereas the migratory function of cutaneous dendritic cells and hapten-specific cell proliferation were not disturbed. Thioredoxin-transgenic mice had also an attenuated inflammatory response to croton oil. These findings suggest that thioredoxin prevents skin inflammatory responses and could be a suitable candidate for the treatment of contact dermatitis. PMID- 19186997 TI - Biological redox switches. AB - Research over the last decade has substantially advanced our understanding of cellular redox chemistry and introduced new terms to redox signaling and biological redox switches. It emerges that redox switching affects a plethora of biological processes and seems to be necessary to counterbalance oxidative stress and guarantee cellular survival in oxidative conditions. Despite intensive studies, the mechanisms of redox switching and its consequences are poorly understood, which points towards an extremely complex nature of these phenomena. Indeed, in contrast to classical signalling cascades, cellular redox signaling seems to affect the whole cellular redox environment and a large number of different redox switches. A majority of biological redox switches rely on the oxidation of thiol group(s) of cysteine residue(s); however, thiolates also bind transition metal ions like Zn(II), Cu(I), and iron, and these metal-thiolate motifs also function as redox switches. It follows that a deeper understanding of redox signalling and redox switches could be achieved by a multidisciplinary approach combining advances in the redox chemistry of sulphur, the chemistry of reactive oxygen and nitrogen species, as well as the bioinorganic chemistry of metal complexes. Many of these aspects are reviewed in the current forum issue on biological redox switches with the aim to promote the understanding of cellular redox phenomena at system biology level. PMID- 19186998 TI - SUMO proteases: redox regulation and biological consequences. AB - Small-ubiquitin modifier (SUMO) has emerged as a novel modification system that governs the activities of a wide spectrum of protein substrates. SUMO-specific proteases (SENP) are of particular interest, as they are responsible for both the maturation of SUMO precursors and for their deconjugation. The interruption of SENPs has been implicated in embryonic defects and carcinoma cells, indicating that a proper balance of SUMO conjugation and deconjugation is crucial. Recent advances in molecular and cellular biology have highlighted the distinct subcellular localization, and endopeptidase and isopeptidase activities of SENPs, suggesting that they are nonredundant. A better understanding of the molecular basis of SUMO recognition and hydrolytic cleavage has been obtained from the crystal structures of SENP-substrate complexes. While a number of proteomic studies have shown an upregulation of sumoylation, attention is now increasingly being directed towards the regulatory mechanism of sumoylation, in particular the oxidative effect. Findings on the oxidation-induced intermolecular disulfide of E1-E2 ligases and SENP1/2 have improved our understanding of the mechanism by which modification is switched up or down. More intriguingly, a growing body of evidence suggests that sumoylation cross-talks with other modifications, and that the upstream and downstream signaling pathway is co-regulated by more than one modifier. PMID- 19186999 TI - Alpha-tocopherol is ineffective in preventing the decomposition of preformed lipid peroxides and may promote the accumulation of toxic aldehydes: a potential explanation for the failure of antioxidants to affect human atherosclerosis. AB - The decomposition of peroxidized lipids of low-density lipoprotein (LDL) has been suggested to be involved in atherosclerosis. In this study, an in vitro system with 13-hydroperoxylinoleic acid (13-HPODE) was used to determine the effects of antioxidants on its decomposition. Decomposition of 13-HPODE was not affected by alpha-tocopherol, several other antioxidants, or antioxidant enzymes. Moreover, the inclusion of alpha-tocopherol during the decomposition of 13-HPODE resulted in an accumulation of aldehydes. Further oxidation of aldehydes to carboxylic acids by a number of oxidases was prevented by alpha-tocopherol. Conversely, the formation of carboxylic acids may be conducive to plaque stabilization via immunomodulation, rapid degradation, and by calcium sequestration. Thus, the inhibition of formation of carboxylic acids could be a serious deleterious effect of antioxidant treatment. In contrast, alpha-keto acids, like pyruvic acid, promoted the conversion of 13-HPODE to 13-hydroxylinoleic acid (13-HODE) by readily undergoing decarboxylation into acetate. These observations suggest that agents that promote the reduction of lipid peroxides into lipid hydroxides could be far more effective in treating cardiovascular diseases as opposed alpha tocopherol-like antioxidants that could affect additional steps in the oxidation cascade. PMID- 19187000 TI - Biphasic, bidirectional regulation of NF-kappaB by endoplasmic reticulum stress. AB - Endoplasmic reticulum (ER) stress induces an adaptive program called the unfolded protein response (UPR), which affects activity of an array of kinases and transcription factors. Previous reports provided evidence for activation of nuclear factor-kappaB (NF-kappaB), the major transcription factor regulating inflammatory processes, by ER stress. However, recent investigation also suggested that preceding ER stress suppresses activation of NF-kappaB by subsequent exposure to inflammatory stimuli. Although ER stress induces activation of NF-kappaB in the early phase, consequent UPR may inhibit NF-kappaB dependent cellular activation in the later phase. This article summarizes current knowledge on potential mechanisms underlying the biphasic, bidirectional regulation of NF-kappaB by ER stress. PMID- 19187002 TI - Thioredoxin-1 and endothelial cell aging: role in cardiovascular diseases. AB - The thioredoxin-1 (Trx-1) system consists of two oxidoreductases, thioredoxin reductase and Trx-1. Trx-1 is a ubiquitously expressed oxidoreductase. The cellular functions of Trx-1 are wide range. They include protein disulfide reduction, DNA synthesis, protection from apoptosis, redox regulation of a variety of proteins, transcription factors and reduction of H(2)O(2), respectively. This review will first focus on the essential role for Trx-1 in different cardiovascular cells, namely smooth muscle cells, endothelial cells, and cardiomyocytes. Thereby, the review will demonstrate the predominant role of Trx-1 to limit oxidative stress directly due to reactive oxygen species scavenging and by protein-protein interaction with key signaling molecules. Second, this review will highlight the role of Trx-1 in cardiovascular aging, focusing on its importance on shear stress and the profound changes with age. Finally, the review will focus on important in vivo studies showing a protective role of Trx-1 in different cardiovascular diseases. Thus, the Trx system and Trx 1 could be important future targets to develop clinical therapies for cardiovascular disorders. PMID- 19187001 TI - Redox modifier genes and pathways in amyotrophic lateral sclerosis. AB - Enhanced redox-stress caused by neuroinflammation, mitochondria, and NADPH oxidases has been hypothesized to play critical roles in disease progression of amyotrophic lateral sclerosis (ALS). However, distinguishing whether the redox stress observed in ALS is due to a primary defect in cellular reactive oxygen species metabolism/catabolism, or is a secondary consequence of neuroinflammation, has been difficult and the issue remains a matter of debate. Emerging evidence suggests that defects in genes that regulate NADPH oxidases may account for at least some forms of ALS. NADPH oxidases are key signaling complexes that influence cellular responses to growth factors and cytokines. In this context, NADPH oxidase-derived reactive oxygen species exert spatial control over the redox-dependent activation of certain pro-inflammatory receptors. Understanding the biology of how NADPH oxidases control cell signaling may help to clarify how genetic determinants of ALS lead to dysregulated pro-inflammatory signaling. This review provides a framework for understanding endosomal signaling through NADPH oxidases and potential mechanisms whereby gene defects in various forms of ALS may influence this cellular process and lead to motor neuron degeneration. Lastly, this review discusses past and current efforts to treat ALS using antioxidant therapies, as well as the limitations and advantages of each of these approaches. PMID- 19187003 TI - Therapeutic angiogenesis in diabetes and hypercholesterolemia: influence of oxidative stress. AB - Despite significant improvements in the medical, percutaneous, and surgical management, numerous patients are first seen with non-revascularizable coronary artery disease (CAD). The growth of new blood vessels to improve myocardial perfusion (i.e., therapeutic angiogenesis) is an attractive treatment option for these patients. However, the successes of angiogenic therapy, observed in preclinical studies, have not been realized in clinical trials. Increasing evidence suggests that this discrepancy between animal and human studies may be due to the nature of the substrate, or the molecular and cellular environment within which the angiogenic agent acts. Antiangiogenic influences, including endothelial dysfunction, hypercholesterolemia, and diabetes, are present in virtually all patients with advanced CAD. Recent studies have better characterized the abnormalities associated with these disease states, providing novel targets for intervention. These substrate-modifying interventions can potentially enhance the response to protein-, gene-, or cell-based angiogenic therapy. In this review, we discuss key aspects of the angiogenic process and the pathophysiologic and molecular mechanisms that contribute to an impaired angiogenic response in the setting of endothelial dysfunction, hypercholesterolemia, and diabetes, with a focus on the role of oxidative stress. Last, we briefly explore substrate modifying agents that have been evaluated in preclinical and clinical studies to improve the angiogenic response. PMID- 19187006 TI - Different redox states in malignant and nonmalignant esophageal epithelial cells and differential cytotoxic responses to bile acid and honokiol. AB - Esophageal adenocarcinoma (EAC) is a highly lethal cancer in western countries. EAC cells are believed to develop from esophageal epithelial cells through complex transformation processes involving inflammation and oxidative stress. The purpose of this study was to compare the redox status of malignant and nonmalignant esophageal epithelial cells and to test their responses to bile acid induced oxidative stress and to treatment with honokiol (HNK), a natural product with anticancer activity. We demonstrated that esophageal adenocarcinoma cells express significantly higher levels of antioxidant molecules and were resistant to reactive oxygen species (ROS) stress induced by bile acid, but were sensitive to the cytotoxic action of HNK. Mechanistic study showed that HNK caused cancer cell death by disruption of mitochondrial transmembrane potential and was correlated with cyclophilin D (CypD) expression. Inhibition of CypD by cyclosporin A or abrogation of its expression by siRNA significantly suppressed the cytotoxicity of HNK, suggesting that CypD may be a key molecule that mediates the cytotoxicity. Our study suggests that the high antioxidant capacity in EAC cells confers on them the ability to survive the oxidative microenvironment in the reflux esophagus, and that HNK is a promising compound to kill the transformed cells preferentially. PMID- 19187005 TI - Proinflammatory stem cell signaling in cardiac ischemia. AB - Cardiovascular disease remains a leading cause of mortality in developed nations, despite continued advancement in modern therapy. Progenitor and stem cell-based therapy is a novel treatment for cardiovascular disease, and modest benefits in cardiac recovery have been achieved in small clinical trials. This therapeutic modality remains challenged by limitations of low donor-cell survival rates, transient recovery of cardiac function, and the technical difficulty of applying directed cell therapy. Understanding the signaling mechanisms involved in the stem cell response to ischemia has revealed opportunities to modify directly aspects of these pathways to improve their cardioprotective abilities. This review highlights general considerations of stem cell therapy for cardiac disease, reviews the major proinflammatory signaling pathways of mesenchymal stem cells, and reviews ex vivo modifications of stem cells based on these pathways. PMID- 19187009 TI - Characterization and optimization of a red-shifted fluorescence polarization ADP detection assay. AB - ATP depletion and ADP formation are generic detection methods used for the identification of kinase and other ATP-utilizing enzyme inhibitors in high throughput screening campaigns. However, the most widely used nucleotide detection approaches require high ATP consumption rates or involve the use of coupling enzymes, which can complicate the selection of lead compounds. As an alternative, we have developed the Transcreener (BellBrook Labs, Madison, WI) platform, which relies on the direct immunodetection of nucleotides. Here we describe the development of antibodies with >100-fold selectivity for ADP versus ATP, which enable robust detection of initial velocity rates (Z' > 0.7 at 10% substrate consumption) at ATP concentrations ranging from 0.1 microM to 1,000 microM in a competitive fluorescence polarization (FP) immunoassay. Competitive binding experiments indicate similar affinities for other nucleotide diphosphates, including 2' -deoxy ADP, GDP, and UDP. The antibody-tracer complex and the red-shifted, ratiometric FP signal are stable for at least 24 h at room temperature, providing suitable conditions for high-throughput screening. A method for calculating a kinase ATP Km with this FP immunoassay is also presented. The Transcreener ADP assay provides a simple, generic assay platform for inhibitor screening and selectivity profiling that can be used for any ADP generating enzyme. PMID- 19187010 TI - WebFlow: a software package for high-throughput analysis of flow cytometry data. AB - Flow cytometry has emerged as a powerful tool for quantitative, single-cell analysis of both surface markers and intracellular antigens, including phosphoproteins and kinase signaling cascades, with the flexibility to process hundreds of samples in multiwell plate format. Quantitative flow cytometric analysis is being applied in many areas of biology, from the study of immunology in animal models or human patients to high-content drug screening of pharmacologically active compounds. However, these experiments generate thousands of data points per sample, each with multiple measured parameters, leading to data management and analysis challenges. We developed WebFlow (http://webflow.stanford.edu), a web server-based software package to manage, analyze, and visualize data from flow cytometry experiments. WebFlow is accessible via standard web browsers and does not require users to install software on their personal computers. The software enables plate-based annotation of large data sets, which provides the basis for exploratory data analysis tools and rapid visualization of multiple different parameters. These tools include custom user-defined statistics to normalize data to other wells or other channels, as well as interactive, user-selectable heat maps for viewing the underlying single-cell data. The web-based approach of WebFlow allows for sharing of data with collaborators or the general public. WebFlow provides a novel platform for quantitative analysis of flow cytometric data from high-throughput drug screening or disease profiling experiments. PMID- 19187011 TI - Laparoscopic cystoprostatectomy for radiation-induced hemorrhagic cystitis. AB - BACKGROUND AND PURPOSE: Hemorrhagic cystitis after radiotherapy is a relatively rare event, usually occurring several years after radiation treatment. It can be relentlessly progressive, and treatment options are usually suboptimal. No management strategy is fully successful, and a stepwise progression in treatment intensity is often necessary with cystectomy and urinary diversion being the final management option. PATIENTS AND METHODS: Laparoscopic cystoprostatectomy with minilaparotomy ileal conduit urinary diversion was successfully performed in three patients with severe refractory hemorrhagic cystitis secondary to radiation treatment for prostate cancer. More conservative management had failed in all three patients with continued severe and life-threatening bleeding. RESULTS: No intraoperative complications were encountered in spite of the obliteration of tissue planes after radiation treatment. Mean operative time was 4 hours and 45 minutes, including the urinary diversion. Blood loss ranged from 50 to 200 mL. Postoperative recovery was uneventful with the exception of prolonged ileus in one patient, which resolved conservatively. CONCLUSION: Radiation treatment for prostate cancer is not an absolute contraindication for laparoscopic cystoprostatectomy, which can still be safely performed in some of these patients. Extension of this experience to patients with malignant disease remains to be evaluated. PMID- 19187004 TI - Mitochondrial reactive oxygen species production in excitable cells: modulators of mitochondrial and cell function. AB - The mitochondrion is a major source of reactive oxygen species (ROS). Superoxide (O(2)(*-)) is generated under specific bioenergetic conditions at several sites within the electron-transport system; most is converted to H(2)O(2) inside and outside the mitochondrial matrix by superoxide dismutases. H(2)O(2) is a major chemical messenger that, in low amounts and with its products, physiologically modulates cell function. The redox state and ROS scavengers largely control the emission (generation scavenging) of O(2)(*-). Cell ischemia, hypoxia, or toxins can result in excess O(2)(*-) production when the redox state is altered and the ROS scavenger systems are overwhelmed. Too much H(2)O(2) can combine with Fe(2+) complexes to form reactive ferryl species (e.g., Fe(IV) = O(*)). In the presence of nitric oxide (NO(*)), O(2)(*-) forms the reactant peroxynitrite (ONOO(-)), and ONOOH-induced nitrosylation of proteins, DNA, and lipids can modify their structure and function. An initial increase in ROS can cause an even greater increase in ROS and allow excess mitochondrial Ca(2+) entry, both of which are factors that induce cell apoptosis and necrosis. Approaches to reduce excess O(2)(*-) emission include selectively boosting the antioxidant capacity, uncoupling of oxidative phosphorylation to reduce generation of O(2)(*-) by inducing proton leak, and reversibly inhibiting electron transport. Mitochondrial cation channels and exchangers function to maintain matrix homeostasis and likely play a role in modulating mitochondrial function, in part by regulating O(2)(*-) generation. Cell-signaling pathways induced physiologically by ROS include effects on thiol groups and disulfide linkages to modify posttranslationally protein structure to activate/inactivate specific kinase/phosphatase pathways. Hypoxia-inducible factors that stimulate a cascade of gene transcription may be mediated physiologically by ROS. Our knowledge of the role played by ROS and their scavenging systems in modulation of cell function and cell death has grown exponentially over the past few years, but we are still limited in how to apply this knowledge to develop its full therapeutic potential. PMID- 19187012 TI - Case report: loss of hernia mesh after simultaneous laparoscopic extraperitoneal lymphadenectomy, radical prostatectomy, and hernioplasty. AB - We present a case of prosthetic mesh hernia repair of a unilateral inguinal hernia following laparoscopic extraperitoneal pelvic lymph node dissection and radical prostatectomy. After an uneventful intraoperative and early postoperative period, the patient developed a lymphocele. This resulted in the detachment of the mesh from the abdominal wall, which necessitated its removal. PMID- 19187013 TI - Pediatric extracorporeal shockwave lithotripsy: its efficiency at various locations in the upper tract. AB - BACKGROUND: Several reports demonstrated the effectiveness of extracorporeal shockwave lithotripsy (SWL) in the management of pediatric upper tract urolithiasis. The majority of these studies used the stone-free rate as the main indicator for treatment success. Efficacy Quotient (EQ) is an important outcome measure because it not only takes into consideration the stone-free rate, but also includes other undesirable outcomes that increase patient morbidity and utilizes hospital resources. Nevertheless, very few studies reported on the EQ of pediatric SWL at various locations in the upper tract. METHODS: We retrospectively reviewed the records of 84 children (103 urinary calculi) who were treated with SWL in our institution between 1995 and 2005. RESULTS: There were 84 renal (pelvis, n = 27; upper calyx, n = 13; middle calyx, n = 8; lower calyx, n = 36) and 19 ureteric (upper third, n = 10; middle third, n = 5; lower third n = 4) stones. EQ for renal pelvic stones was 67% compared to 48% for caliceal stones (upper calyx: 52%, middle calyx: 46% and lower calyx: 47%). Overall, the EQ for renal stone was 54% and it was 59% for ureteric stones. This efficiency in the ureter was achieved at the expense of a higher number of shock waves delivered (3606 vs. 2592, P < 0.01). Overall, the stone-free rate was 82% (renal, 81%; ureteric calculi 84%). The only factor that affected the stone-free rate was the size of the stone (33% in stones >2 cm and 87% in stones <2 cm (P < 0.01). The overall retreatment rate was 41% (renal, 40% and ureteric 42%). However, the number of total treatments ranged from 1 to 5 sessions in renal stones and 1 to 2 sessions in ureteric stones. Auxiliary procedures were performed in 9 patients (stenting, n = 3; ureteroscopy, n = 4 and percutaneous nephrolithotomy, n = 2). CONCLUSIONS: Pediatric SWL appears to be more efficient in the renal pelvic stones compared to caliceal stones. The efficiency of SWL in the ureter is achieved with a higher number of shock waves delivered. A discussion on the possible future modifications of the definition of EQ, especially in children, is also provided. PMID- 19187014 TI - Determination of the most efficient temperature for radiofrequency ablation of renal cells: a prospective study in dogs. AB - BACKGROUND AND PURPOSE: Radiofrequency (RF) ablation of renal tumors is a major technique for tumor cell destruction while preserving healthy renal parenchyma. There is no consensus in the literature regarding the optimal temperature, impedance, and time for RF application for effective cell destruction. This study investigated two variables while keeping time unchanged: Temperature for RF cell destruction and tissue impedance in dog kidneys. MATERIALS AND METHODS: Sixteen dogs had renal punctures through videolaparoscopy for RF interstitial tissue ablation. A RF generator was applied for 10 minutes to the dog's kidney at different target temperatures: 80 degrees C, 90 degrees C, and 100 degrees C. On postoperative day 14, the animals were sacrificed and nephrectomized. All lesions were macroscopically and microscopically examined. The bioelectrical impedance was evaluated at three different temperatures. RESULTS: Renal injuries were wider and deeper at 90 degrees C (P < 0.001), and they were similar at 80 degrees C and 100 degrees C. The bioelectrical impedance was lower at 90 degrees C than at the temperatures of 80 degrees C and 100 degrees C (P < 0.001). Viable cells in the RF ablation tissue area were not found in the microscopic examination. CONCLUSION: The most effective cell destruction in terms of width and depth was achieved at 90 degrees C, which was also the optimal temperature for tissue impedance. RF ablation of renal cells eliminated all viable cells. PMID- 19187015 TI - Combined retrograde flexible ureteroscopic lithotripsy with holmium YAG laser for renal calculi associated with ipsilateral ureteral stones. AB - PURPOSE: The purpose of this study was to evaluate the effectiveness of combined ureteroscopic holmium YAG lithotripsy for renal calculi associated with ipsilateral ureteral stones. MATERIALS AND METHODS: Between August 2002 and March 2007, retrograde flexible ureteroscopic stone treatment was attempted in 351 cases. Indication for treatment was concurrent symptomatic ureteral stones in 63 patients (group I). Additional operative time and perioperative complication rates were compared to a group of 39 patients submitted to ureteroscopic treatment for ureteral calculi exclusively (group II). RESULTS: Mean ureteral stone size was 8.0 +/- 2.6 mm and 8.1 +/- 3.4 mm for groups I and II, respectively. Mean operative time for group I was 67.9 +/- 29.5 minutes and for group 2 was 49.3 +/- 13.2 minutes (p < 0.001). Flexible ureteroscopic therapy for renal calculi increased 18 minutes in the mean operative time. The overall complication rate was 3.1% and 2.5% for groups I and II, respectively (p = 0.87). Mean renal stone size was 10.7 +/- 6.4 mm, overall stone free rate in group I was 81%. However, considering only patients with renal stones smaller than 15 mm, the stone free rate was 88%. Successful treatment occurred in 81% of patients presenting lower pole stones, but only 76% of patients with multiple renal stones became stone free. As expected, stone free rate showed a significant negative correlation with renal stone size (p = 0.03; r = -0.36). Logistic regression model indicated an independent association of renal stones smaller than 15 mm and stone free rate (OR = 13.5; p = 0.01). CONCLUSION: Combined ureteroscopic treatment for ureteral and ipsilateral renal calculi is a safe and attractive option for patients presenting for symptomatic ureteral stone and ipsilateral renal calculi smaller than 15 mm. PMID- 19187016 TI - Effects of laser on the synovial fluid in the inflammatory process of the knee joint of the rabbit. AB - OBJECTIVE: The purpose of this study was to evaluate the effects of low-level laser (LLL) energy on the clinical signs of inflammation and the cellular composition of synovial fluid (SF) in the inflamed knee of the rabbit. BACKGROUND DATA: There are few findings related to the effects of LLL on SF in inflammatory processes and there is little knowledge about the optimal parameters for reducing joint inflammation. MATERIALS AND METHODS: Inflammation in the right knee of 36 rabbits was induced by intracapsular injection (0.2 mL) of Terebinthina commun (Tc). The animals were randomly assigned to three groups: acute experimental group (AEG), chronic experimental group (CEG), and control group (CG), which only received Tc. Each group was divided in two subgroups of six animals each. The AEG and CEG groups began to receive laser treatment 2 and 5 d after the induction of inflammation, respectively. Laser irradiation at a wavelength of 830 nm, power output of 77 mW, and power density of 27.5 W/cm(2) was applied daily for 7 d for either 0.12 sec or 0.32 sec, resulting in doses of 3.4 J/cm(2) and 8 J/cm(2), respectively. Body mass, joint perimeter, joint temperature, and the morphology of the SF were analyzed. RESULTS: There was no statistically significant differences between groups in the body mass, joint perimeter, and SF morphology. CONCLUSION: Laser irradiation with the selected parameters produced only a few subtle differences in the inflammatory signs and the SF. The lack of effects may have been due to the short irradiation time. PMID- 19187017 TI - Characterization of Canadian cigarettes using multi-stable isotope analysis by gas chromatography-isotope ratio mass spectrometry. AB - A reliable method for measuring (15)N/(14)N and (2)H/(1)H ratios of nicotine in cigarette tobacco has been developed. It involves a simple procedure for extracting nicotine from tobacco using methanol. The extract is directly analyzed on a gas chromatography isotope ratio mass spectrometer (GC-IRMS). The method is reproducible with 4% RSD for delta(2)H and delta(15)N. Brand name cigarettes manufactured in Canada (N = 47) and in China (N = 23) have been analyzed. The results show that nicotine from Canadian cigarettes has a higher (2)H/(1)H ratio and a lower (15)N/(14)N ratio than the Chinese cigarettes. The delta(2)H values for Canadian cigarettes range from -232.7 per thousand to -203.4 per thousand with an average of -222.1 per thousand; the delta(2)H values for Chinese cigarettes range from -262.6 per thousand to -219.9 per thousand with an average of -243.8 per thousand. The delta(15)N values for Canadian cigarettes range from 7.7 per thousand to -6.3 per thousand with an average of -7.1 per thousand; the delta(15)N values for Chinese cigarettes range from -7.6 per thousand to -5.7 per thousand with an average of -6.3 per thousand. The combined measurements of (2)H/(1)H and (15)N/(14)N have been shown to be useful in identifying counterfeits of Canadian cigarettes analyzed in this study. PMID- 19187018 TI - Cellular lipid metabolism is influenced by the coordination environment of copper. AB - Copper ions are vital to human health, and mis-trafficking of them can result in many diseases including Wilson's, Menkes', and Alzheimer's diseases. Coherent anti-Stokes Raman scattering (CARS) microscopy can be used to observe changes in lipid phenotype in a noninvasive manner and is employed here to show that copper accumulation in hepatic cells results in rapid changes in lipid storage and lipid droplet density. The increase in lipid storage is dependent on the coordination environment of the copper to which the cells are exposed and changes in toxicity, lipid phenotype, and rate of copper accumulation upon treatment vary using different Cu species. PMID- 19187019 TI - Carbohydrate dependent targeting of cancer cells by bleomycin-microbubble conjugates. AB - Biotinylated bleomycin A(5) was attached to streptavidin-derivatized microbubbles, and a solution containing the conjugate was passed over a monolayer of cultured MCF-7 cells. The bleomycin-derivatized microbubbles adhered to the MCF-7 cells, and the association could be monitored by the use of a microscope. Three other cancer cell lines gave similar results. The bleomycin-microbubble conjugate did not bind to a normal breast cell line (MCF-10A) or to the matched noncancer cell lines corresponding to the other cancer cell lines targeted by bleomycin. No binding to any tested cell line was observed when the microbubbles lacked conjugated bleomycin A(5) or when the microbubble contained a bleomycin A(5) analogue lacking the carbohydrate moiety. PMID- 19187020 TI - Various methods for determination of the degree of N-acetylation of chitin and chitosan: a review. AB - Chitin, chitosan, and their derivatives have been identified as versatile biopolymers for a broad range of agriculture and food applications. Up to now, several methods have been developed to determine degree of N-acetylation, DA, for chitin and chitosan. In this article, an effort has been made to review the available literature information on the DA determination. These methods are classified into three categories: (1) spectroscopy (IR, (1)H NMR, (13)C NMR, (15)N NMR, and UV); (2) conventional (various types of titration, conductometry, potentiometry, ninhydrin assay, adsorption of free amino groups of chitosan by pictric acid); (3) destructive (elemental analysis, acid or enzymatic hydrolysis of chitin/chitosan and followed by the DA measurement by colorimetry or high performance liquid chromatography, pyrolysis-gas chromatography, and thermal analysis using differential scanning calorimetry) methods. These methods have been compared for their performances and limitations as well as their advantages and disadvantages. The use of IR and NMR spectroscopy methods provides a number of advantages. They do not need long-term procedures to prepare samples, and they provide information on the chemical structure. (1)H NMR and UV techniques are more sensitive than IR, (13)C NMR, and (15)N NMR spectroscopy. The IR technique is mostly used for a qualitative evaluation and comparison studies. Conventional methods are not applicable for highly acetylated chitin. The results of the latter methods are affected by ionic strength of the solvent, pH, and temperature of solution. In destructive methods, longer times are needed for the measurements compared to spectroscopy and conventional methods, but they are applicable for the entire range of the DA. PMID- 19187021 TI - Biochemical characterization of MODY2 glucokinase variants V62M and G72R reveals reduced enzymatic activities relative to wild type. AB - The glucokinase V62M and G72R mutations are naturally occurring and known to associate with hyperglycemia in humans. Structurally, V62 and G72 residues are located in close proximity to the allosteric site where hypoglycemia-linked activating mutations are clustered. To address the mechanism by which these variants alter the physiological phenotype, we characterized the biochemical and biophysical properties of the enzymes. Recombinant proteins were purified without affinity tags, and their steady-state kinetics and glucose binding affinities were determined. Both enzymes showed reduced rates of turnover (k(cat)) and reduced glucose affinity (i.e., increased K(0.5) and K(D) values). Their thermal stability did not largely differ from that of wild-type glucokinase. However, V62M and G72R lost the stabilizing protein interactions with glucokinase regulatory protein, which may contribute to lower activity in vivo. Both mutants were subject to activation by small molecule activators. In conclusion, the decreased enzyme activities of V62M and G72R observed in this study are consistent with the hyperglycemic phenotype. PMID- 19187022 TI - Coffee and green tea as a large source of antioxidant polyphenols in the Japanese population. AB - Food and beverages rich in polyphenols with antioxidant activity are highlighted as a potential factor for risk reduction of lifestyle related diseases. This study was conducted to elucidate total polyphenol consumption from beverages in Japanese people. Total polyphenol (TP) contents in beverages were measured using a modified Folin-Ciocalteu method removing the interference of reduced sugars by using reverse-phase column chromatography. A beverage consumption survey was conducted in the Tokyo and Osaka areas in 2004. Randomly selected male and female subjects (10-59 years old, n = 8768) recorded the amounts and types of all nonalcoholic beverages consumed in a week. Concentration of TP in coffee, green tea, black tea, Oolong tea, barley tea, fruit juice, tomato/vegetable juice, and cocoa drinks were at 200, 115, 96, 39, 9, 34, 69, and 62 mg/100 mL, respectively. Total consumption of beverages in a Japanese population was 1.11 +/- 0.51 L/day, and TP contents from beverages was 853 +/- 512 mg/day. Coffee and green tea shared 50% and 34% of TP consumption in beverages, respectively, and contribution of each of the other beverages was less than 10%. TP contents in 20 major vegetables and 5 fruits were 0-49 mg and 2-55 mg/100 g, respectively. Antioxidant activities, Cu reducing power, and scavenging activities for DPPH and superoxide, of those samples correlated to the TP contents (p < 0.001). Beverages, especially coffee, contributed to a large share of the consumption of polyphenols, as antioxidants, in the Japanese diet. PMID- 19187023 TI - Nanoporous crystals of calixarene/porphyrin supramolecular complex functionalized by diffusion and coordination of metal ions. AB - A highly nanoporous material has been obtained by self-assembly of calixarene and porphyrin building blocks. This supramolecular zeolite-like structure was successively functionalized by diffusion and coordination of metal ions to form a new bifunctionalized nanoporous material containing a porphyrinic pigment together with a metal center. PMID- 19187024 TI - Effects of long-term frozen storage on electrophoretic patterns, immunoreactivity, and pepsin in vitro digestibility of soybean (Glycine max L.) proteins. AB - Soybean flours stored for 20 years at -20 degrees C retained protein polypeptide profile integrity. Proteins in stored soybean flours retained their immunoreactivity. Long-term frozen storage of seed flours at -20 degrees C did not adversely affect seed protein in vitro pepsin digestibility. PMID- 19187026 TI - Population-based study of invasive disease due to beta-hemolytic streptococci of groups other than A and B. AB - BACKGROUND: Beta-hemolytic streptococci of groups other than A and B (NABS) are increasingly recognized as causes of clinically significant disease, but precise information about this heterogeneous group is lacking. We report the incidence of NABS infection and describe the epidemiologic and clinical characteristics. METHODS: Active, population-based surveillance for invasive NABS was performed over a 2-year period in the 8-county metropolitan Atlanta, Georgia, area and the 3-county San Francisco Bay, California, area. Clinical records were reviewed, and available isolates were sent to the Centers for Disease Control and Prevention (Atlanta) for additional microbiologic characterization. Incidences were calculated using year-appropriate US Census Bureau data. RESULTS: A total of 489 cases of invasive NABS infection were identified (3.2 cases per 100,000 population). The median age of patients was 55 years; 64% of patients were males, and 87% had underlying diseases. The incidence was higher among black persons than white persons (4.0 vs. 2.5 cases per 100,000 population; P < .01) and increased with age among all races. Infections were community acquired in 416 cases (85%). Among the 450 patients (94%) with NABS infection who were hospitalized, 55 (12%) died. Of 266 isolates (54%) speciated at the Centers for Disease Control and Prevention, 212 (80%) were Streptococcus dysgalactiae subspecies equisimilis; 46 (17%) were members of the Streptococcus anginosus group. S. dysgalactiae subspecies equisimilis primarily presented as skin and soft-tissue infection in older patients, whereas individuals with invasive S. anginosus group infections were more likely to be younger patients with intra abdominal infections. CONCLUSIONS: NABS comprise multiple distinct species that cause a significant number of community-acquired invasive infections. Clinical manifestations differ by species. Thus, speciation of invasive NABS may be warranted in clinical settings. PMID- 19187027 TI - Clinical features and molecular epidemiology of CMY-type beta-lactamase-producing Escherichia coli. AB - BACKGROUND: Knowledge of the clinical features of infections caused by Escherichia coli strains that produce plasmid-mediated AmpC beta-lactamase is limited. Of the several groups of plasmid-mediated AmpC beta-lactamases, CMY-type beta-lactamase is the most common in the United States. METHODS: We prospectively identified patients infected or colonized with E. coli strains that produce CMY type beta-lactamase, and we collected clinical data over a 7-month period. A retrospective cohort study was performed to identify features associated with these cases. Patients with extended-spectrum beta-lactamase-producing E. coli were used as a control group. Pulsed-field gel electrophoresis, plasmid analysis, and phylogenetic typing were performed. RESULTS: Twenty-two patients with infection or colonization due to CMY-type beta-lactamase-producing E. coli and 25 patients with infection or colonization due to extended-spectrum beta-lactamase producing E. coli were identified. The demographic characteristics of the patients were similar in both cohorts. Patients with CMY-type beta-lactamase producing E. coli were significantly more likely to have symptomatic infection than were patients with extended-spectrum beta-lactamase-producing E. coli (P = .028). The CMY-type beta-lactamase was identified as CMY-2 or its variants. Ninety-four percent of the CMY-type beta-lactamase-producing isolates belonged to E. coli phylogenetic groups B2 and D, which are associated with virulence. Many of the isolates shared similar plasmid profiles, whereas the pulsed-field gel electrophoresis profiles were diverse. Co-resistance to non-beta-lactam antimicrobials was common. CONCLUSION: In Pittsburgh, Pennsylvania, CMY-type beta lactamase-producing E. coli strains are almost as common as extended-spectrum beta-lactamase-producing E. coli strains, and they cause symptomatic infection in the majority of cases. PMID- 19187029 TI - Association of single-nucleotide polymorphism 3 and c.553G>T of APOA5 with hypertriglyceridemia after treatment with highly active antiretroviral therapy containing protease inhibitors in hiv-infected individuals in Taiwan. AB - We investigated the relationship between hypertriglyceridemia and the single nucleotide polymorphisms (SNPs) on APOA5 in human immunodeficiency virus (HIV) infected patients receiving highly active antiretroviral therapy (HAART) in Taiwan. Receipt of protease inhibitor-based HAART, high baseline triglyceride levels, and carriage of APOA5 SNP3 or c.553G>T variants or APOA5 SNP1T/SNP2G/SNP3C/c.553T haplotype were statistically significantly associated with development of extreme hypertriglyceridemia (triglyceride level, >500 mg/dL). PMID- 19187028 TI - Lumbar puncture in HIV-infected patients with syphilis and no neurologic symptoms. AB - BACKGROUND: The decision to perform lumbar puncture in patients with asymptomatic human immunodeficiency virus (HIV) infection and syphilis is controversial. The Centers for Disease Control and Prevention recommend certain criteria that warrant lumbar puncture. Here, we assess the performance of these criteria for detecting asymptomatic neurosyphilis (ANS). METHODS: Eligible subjects consisted of all patients with concurrent HIV infection and syphilis in a prospective clinical cohort who had no neurologic symptoms at the time of lumbar puncture. We retrospectively applied different stratification criteria to calculate the performance of lumbar puncture in detecting ANS: (1) lumbar puncture in patients with late latent syphilis or syphilis of an unknown duration, regardless of the CD4 cell count or rapid plasma reagin titer; (2) lumbar puncture if the CD4 cell count was 350 cells/mL and/or the rapid plasma reagin titer was 1:32, regardless of the syphilis stage; and (3) lumbar puncture in the context of serologic nonresponse to syphilis therapy. RESULTS: Two hundred two of 231 patients with syphilis did not have neurologic symptoms. Immediate lumbar puncture was performed for 46 patients, and 10 cases (22%) of ANS were detected. With use of the first criterion, 2 (14%) of 10 cases of ANS in patients with early-stage syphilis would have been missed (sensitivity, 80% [95% confidence interval [CI], 44%-97%]; specificity, 23% (95% confidence interval [CI], 11.4%-40.2%) [corrected] Criterion 2 would not have missed any cases of ANS (sensitivity, 100% [95% CI, 70%-100%]; specificity, 13.2% (95% CI, 4.4%-29.1%) [corrected] but would have required that a lumbar puncture be performed for 88% of patients. Performance of lumbar puncture performed in 13 cases based on serologic nonresponse to syphilis therapy yielded 4 cases (31%) of ANS. CONCLUSIONS: In patients with concurrent HIV infection and syphilis, the use of criteria based on rapid plasma reagin titer and CD4 cell count, instead of stage-based criteria, improved the ability to identify ANS. PMID- 19187030 TI - Active water in protein-protein communication within the membrane: the case of SRII-HtrII signal relay. AB - We detect internal water molecules in a membrane-embedded receptor-transducer complex and demonstrate water structure changes during formation of the signaling state. Time-resolved FTIR spectroscopy reveals stimulus-induced repositioning of one or more structurally active water molecules to a significantly more hydrophobic environment in the signaling state of the sensory rhodopsin II (SRII) transducer (HtrII) complex. These waters, distinct from bound water molecules within the SRII receptor, appear to be in the middle of the transmembrane interface region near the Tyr199(SRII)-Asn74(HtrII) hydrogen bond. We conclude that water potentially plays an important role in the SRII --> HtrII signal transfer mechanism in the membrane's hydrophobic core. PMID- 19187031 TI - The archetype gamma-class carbonic anhydrase (Cam) contains iron when synthesized in vivo. AB - A recombinant protein overproduction system was developed in Methanosarcina acetivorans to facilitate biochemical characterization of oxygen-sensitive metalloenzymes from strictly anaerobic species in the Archaea domain. The system was used to overproduce the archetype of the independently evolved gamma-class carbonic anhydrase. The overproduced enzyme was oxygen sensitive and had full incorporation of iron instead of zinc observed when overproduced in Escherichia coli. This, the first report of in vivo iron incorporation for any carbonic anhydrase, supports the need to reevaluate the role of iron in all classes of carbonic anhydrases derived from anaerobic environments. PMID- 19187033 TI - Paramagnetic 13C and 15N NMR analyses of cyanide- (13C15N-) ligated ferric peroxidases: the push effect, not pull effect, modulates the compound I formation rate. AB - Paramagnetic (13)C and (15)N NMR spectroscopy of heme-bound cyanide ((13)C(15)N) was utilized to quantitatively distinguish the electron donor effect (the push effect) from the proximal histidine and hydrogen-bonding effect (the pull effect) from the distal amino acid residues in cytochrome c peroxidase (CcP), ascorbate peroxidase (APX), lignin peroxidase (LiP), and manganese peroxidase (MnP). Paramagnetic (13)C NMR signals of heme-bound (13)C(15)N of these peroxidases were observed in a wide range, -3501 ppm (CcP), -3563 ppm (APX), -3823 ppm (MnP), and 3826 ppm (LiP), while paramagnetic (15)N NMR signals of those were detected in a narrow range, 574 ppm (ARP), 605 ppm (CcP), 626 ppm (LiP), and 654 ppm (MnP). Detailed analysis, combined with the previous results for horseradish peroxidase and Arthromyces ramosus peroxidase, indicated that the push effect is quite different among these peroxidases while the pull effect is similar. More importantly, a strong correlation between the (13)C NMR shift (the push effect) and the compound I formation rate was observed, indicating that the push effect causes a variation in the compound I formation rate. Comparison of the (13)C and (15)N NMR results of these peroxidases with their crystal structures suggests that the orientation of the proximal imidazole plane to the heme N-Fe-N axis controls the push effect and the compound I formation rate of peroxidase. PMID- 19187032 TI - Accommodation of two diatomic molecules in cytochrome bo: insights into NO reductase activity in terminal oxidases. AB - Bacterial heme-copper terminal oxidases react quickly with NO to form a heme nitrosyl complex, which, in some of these enzymes, can further react with a second NO molecule to produce N(2)O. Previously, we characterized the heme a(3) NO complex formed in cytochrome ba(3) from Thermus thermophilus and the product of its low-temperature illumination. We showed that the photolyzed NO group binds to Cu(B)(I) to form an end-on NO-Cu(B) or a side-on copper-nitrosyl complex, which is likely to represent the binding characteristics of the second NO molecule at the heme-copper active site. Here we present a comparative study with cytochrome bo(3) from Escherichia coli. Both terminal oxidases are shown to catalyze the same two-electron reduction of NO to N(2)O. The EPR and resonance Raman signatures of the heme o(3)-NO complex are comparable to those of the a(3) NO complex. However, low-temperature FTIR experiments reveal that photolysis of the heme o(3)-NO complex does not produce a Cu(B)-nitrosyl complex, but that instead, the NO remains unbound in the active-site cavity. Additional FTIR photolysis experiments on the heme-nitrosyl complexes of these terminal oxidases, in the presence of CO, demonstrate that an [o(3)-NO.OC-Cu(B)] tertiary complex can form in bo(3) but not in ba(3). We assign these differences to a greater iron copper distance in the reduced form of bo(3) compared to that of ba(3). Because this difference in metal-metal distance does not appear to affect the NO reductase activity, our results suggest that the coordination of the second NO to Cu(B) is not an essential step of the reaction mechanism. PMID- 19187035 TI - Characterization of dehaloperoxidase compound ES and its reactivity with trihalophenols. AB - Dehaloperoxidase (DHP), the oxygen transport hemoglobin from the terebellid polychaete Amphitrite ornata, is the first globin identified to possess a biologically relevant peroxidase activity. DHP has been shown to oxidize trihalophenols to dihaloquinones in a dehalogenation reaction that uses hydrogen peroxide as a substrate. Herein, we demonstrate that the first detectable intermediate following the addition of hydrogen peroxide to ferric DHP contains both a ferryl heme and a tyrosyl radical, analogous to Compound ES of cytochrome c peroxidase. Furthermore, we provide a detailed kinetic description for the reaction of preformed DHP Compound ES with the substrate 2,4,6-trichlorophenol and demonstrate the catalytic competency of this intermediate in generating the product 2,4-dichloroquinone. Using rapid-freeze-quench electron paramagnetic resonance spectroscopy, we detected a g approximately 2.0058 signal confirming the presence of a protein radical in DHP Compound ES. In the absence of substrate, DHP Compound ES evolves to a new species, Compound RH, which is functionally unique to dehaloperoxidase. We propose that this intermediate plays a protective role against heme bleaching. While unreactive toward further oxidation, Compound RH can be reduced and subsequently bind dioxygen, generating oxyferrous DHP, which may represent the catalytic link between peroxidase and oxygen transport activities in this bifunctional protein. PMID- 19187034 TI - Characterization of the peroxidase mechanism upon reaction of prostacyclin synthase with peracetic acid. Identification of a tyrosyl radical intermediate. AB - Prostacyclin synthase (PGIS) is a membrane-bound class III cytochrome P450 that catalyzes an isomerization of prostaglandin H(2), an endoperoxide, to prostacyclin. We report here the characterization of the PGIS intermediates in reactions with other peroxides, peracetic acid (PA), and iodosylbenzene. Rapid scan stopped-flow experiments revealed an intermediate with an absorption spectrum similar to that of compound ES (Cpd ES), which is an oxo-ferryl (Fe(IV)O) plus a protein-derived radical. Cpd ES, formed upon reaction with PA, has an X-band (9 GHz) EPR signal of g = 2.0047 and a half-saturation power, P(1/2), of 0.73 mW. High-field (130 GHz) EPR reveals the presence of two species of tyrosyl radicals in Cpd ES with their g-tensor components (g(x), g(y), g(z)) of 2.00970, 2.00433, 2.00211 and 2.00700, 2.00433, 2.00211 at a 1:2 ratio, indicating that one is involved in hydrogen bonding and the other is not. The line width of the g = 2 signal becomes narrower, while its P(1/2) value becomes smaller as the reaction proceeds, indicating migration of the unpaired electron to an alternative site. The rate of electron migration ( approximately 0.2 s(-1)) is similar to that of heme bleaching, suggesting the migration is associated with the enzymatic inactivation. Moreover, a g = 6 signal that is presumably a high spin ferric species emerges after the appearance of the amino acid radical and subsequently decays at a rate comparable to that of enzymatic inactivation. This loss of the g = 6 species thus likely indicates another pathway leading to enzymatic inactivation. The inactivation, however, was prevented by the exogenous reductant guaiacol. The studies of PGIS with PA described herein provide a mechanistic model of a peroxidase reaction catalyzed by the class III cytochromes P450. PMID- 19187036 TI - Circular dichroism spectra and electrophoretic mobility shift assays show that human replication protein A binds and melts intramolecular G-quadruplex structures. AB - Noncanonical DNA structures such as G-quadruplexes might obstruct the binding of hRPA, compromising the accuracy of replication, and be a source of genomic instability. In this study, circular dichroism (CD) and electrophoretic mobility shift assay (EMSA) experiments were used to show that hRPA can bind and melt nontelomeric, intramolecular DNA G-quadruplexes under physiologically germane conditions. EMSA results show that hRPA binds to a 58-mer that includes an embedded quadruplex with an affinity equal to or greater than to nonquadruplex forming 58-mers. Moreover, hRPA binds to a 26-mer purine-rich quadruplex-forming sequence with an affinity indistinguishable from that for binding to the complementary pyrimidine-rich sequence. Under the same conditions, hRPA does not have significant affinity for binding to the duplex formed from the two sequences. Thus, DNA secondary structures can significantly modulate the binding affinity of hRPA over and above its known preference for pyrimidine-rich single stranded sequences, so that at least some intramolecular G-quadruplex structures may not inhibit hRPA binding during DNA replication. CD spectral changes in combination with EMSA titrations suggest that one hRPA heterotrimer is sufficient to form a stable complex with an unfolded 26-mer G-quadruplex prior to the binding of a second hRPA molecule. PMID- 19187038 TI - Preferential glutathione conjugation of a reverse diol epoxide compared to a bay region diol epoxide of phenanthrene in human hepatocytes: relevance to molecular epidemiology studies of glutathione-s-transferase polymorphisms and cancer. AB - Bay region diol epoxides are recognized ultimate carcinogens of polycyclic aromatic hydrocarbons (PAH), and in vitro studies have demonstrated that they can be detoxified by conjugation with glutathione, leading to the widely investigated hypothesis that individuals with low activity forms of glutathione-S-transferases are at higher risk of PAH induced cancer, a hypothesis that has found at most weak support in molecular epidemiology studies. A weakness in this hypothesis was that the mercapturic acids resulting from the conjugation of PAH bay region diol epoxides had never been identified in human urine. We recently analyzed smokers' urine for mercapturic acids derived from phenanthrene, the simplest PAH with a bay region. The only phenanthrene diol epoxide-derived mercapturic acid in smokers' urine was produced from the reverse diol epoxide, anti-phenanthrene-3,4 diol-1,2-epoxide (11), not the bay region diol epoxide, anti-phenanthrene-1,2 diol-3,4-epoxide (10), which does not support the hypothesis noted above. In this study, we extended these results by examining the conjugation of phenanthrene metabolites with glutathione in human hepatocytes. We identified the mercapturic acid N-acetyl-S-(r-4,t-2,3-trihydroxy-1,2,3,4-tetrahydro-c-1-phenanthryl)-L cysteine (14a), (0.33-35.9 pmol/mL at 10 microM 8, 24 h incubation, N = 10) in all incubations with phenanthrene-3,4-diol (8) and the corresponding diol epoxide 11, but no mercapturic acids were detected in incubations with phenanthrene-1,2 diol (7), and only trace amounts were observed in incubations with the corresponding bay region diol epoxide 10. Taken together with our previous results, these studies clearly demonstrate that glutathione conjugation of a reverse diol epoxide of phenanthrene is favored over conjugation of a bay region diol epoxide. Since reverse diol epoxides of PAH are generally weakly or nonmutagenic/carcinogenic, these results, if generalizable to other PAH, do not support the widely held assumption that glutathione-S-transferases are important in the detoxification of PAH in humans. PMID- 19187040 TI - A novel ATM-dependent X-ray-inducible gene is essential for both plant meiosis and gametogenesis. AB - DNA damage in Arabidopsis thaliana seedlings results in upregulation of hundreds of genes. One of the earliest and highest levels of induction is displayed by a previously uncharacterized gene that we have termed X-ray induced 1 (XRI1). Analysis of plants carrying a null xri1 allele revealed two distinct requirements for this gene in plant fertility. XRI1 was important for the post-meiotic stages of pollen development, leading to inviability of xri(-) pollen and abnormal segregation of the mutant allele in heterozygous xri1(+/-) plants. In addition, XRI1 was essential for male and female meiosis, as indicated by the complete sterility of homozygous xri1 mutants due to extensive chromosome fragmentation visible in meiocytes. Abolition of programmed DNA double-strand breaks in a spo11 1 mutant background failed to rescue the DNA fragmentation of xri1 mutants, suggesting that XRI1 functions at an earlier stage than SPO11-1 does. Yeast two hybrid studies identified an interaction between XRI1 and a novel component of the Arabidopsis MND1/AHP2 complex, indicating possible requirements for XRI1 in meiotic DNA repair. PMID- 19187041 TI - Identification of an active LTR retrotransposon in rice. AB - Transposable elements are ubiquitous components of plant genomes. When active, these mobile elements can induce changes in the genome at both the structural and functional levels. Availability of the complete genome sequence for several model plant species provides the opportunity to study TEs in plants at an unprecedented scale. In the case of rice, annotation of the genomic sequence of the variety Nipponbare has revealed that TE-related sequences form more than 25% of its genome. However, most of the elements found are inactive, either because of structural alterations or because they are the target of various silencing pathways. In this paper, we propose a new post-genomic strategy aimed at identifying active TEs. Our approach relies on transcript profiling of TE-related sequences using a tiling microarray. We applied it to a particular class of TEs, the LTR retrotransposons. A transcript profiling assay of rice calli led to identification of a new transpositionally active family, named Lullaby. We provide a complete structural description of this element. We also show that it has recently been active in planta in rice, and discuss its phylogenetic relationships with Tos17, the only other active LTR retrotransposon described so far in the species. PMID- 19187042 TI - CIPK6, a CBL-interacting protein kinase is required for development and salt tolerance in plants. AB - Calcineurin B-like proteins (CBL) and CBL-interacting protein kinases (CIPK) mediate plant responses to a variety of external stresses. Here we report that Arabidopsis CIPK6 is also required for the growth and development of plants. Phenotype of tobacco plants ectopically expressing a homologous gene (CaCIPK6) from the leguminous plant chickpea (Cicer arietinum) indicated its functional conservation. A lesion inAtCIPK6 significantly reduced shoot-to-root and root basipetal auxin transport, and the plants exhibited developmental defects such as fused cotyledons, swollen hypocotyls and compromised lateral root formation, in conjunction with reduced expression of a number of genes involved in auxin transport and abiotic stress response. The Arabidopsis mutant was more sensitive to salt stress compared to wild-type, while overexpression of a constitutively active mutant of CaCIPK6 promoted salt tolerance in transgenic tobacco. Furthermore, tobacco seedlings expressing the constitutively active mutant of CaCIPK6 showed a developed root system, increased basipetal auxin transport and hypersensitivity to auxin. Our results provide evidence for involvement of a CIPK in auxin transport and consequently in root development, as well as in the salt stress response, by regulating the expression of genes. PMID- 19187043 TI - A complementary role for ELF3 and TFL1 in the regulation of flowering time by ambient temperature. AB - Plants regulate their time to flowering by gathering information from the environment. Photoperiod and temperature are among the most important environmental variables. Sub-optimal, but not near-freezing, temperatures regulate flowering through the thermosensory pathway, which overlaps with the autonomous pathway. Here we show that ambient temperature regulates flowering by two genetically distinguishable pathways, one requiring TFL1 and another requiring ELF3. The delay in flowering time observed at lower temperatures was partially suppressed in single elf3 and tfl1 mutants, whereas double elf3 tfl1 mutants were insensitive to temperature. tfl1 mutations abolished the temperature response in cryptochrome mutants that are deficient in photoperiod perception, but not in phyB mutants, which have a constitutive photoperiodic response. In contrast to tfl1, elf3 mutations were able to suppress the temperature response in phyB mutants, but not in cryptochrome mutants. Gene expression profiles revealed that the tfl1 and elf3 effects are due to the activation of different sets of genes, and identified CCA1 and SOC1/AGL20 as being important cross-talk points. Finally, genome-wide gene expression analysis strongly suggests a general and complementary role for ELF3 and TFL1 in temperature signalling. PMID- 19187044 TI - Anesthetic considerations for major burn injury in pediatric patients. AB - Major burn injury remains a significant cause of morbidity and mortality in pediatric patients. With advances in burn care and with the development of experienced multi-disciplinary teams at regionalized burn centers, many children are surviving severe burn injury. As members of the multi-disciplinary care team, anesthesia providers are called upon to care for these critically ill children. These children provide several anesthetic challenges, such as difficult airways, difficult vascular access, fluid and electrolyte imbalances, altered temperature regulation, sepsis, cardiovascular instability, and increased requirements of muscle relaxants and opioids. The anesthesia provider must understand the physiologic derangements that occur with severe burn injury as well as the subsequent anesthetic implications. PMID- 19187045 TI - Healthcare provider-child-parent communication in the preoperative surgical setting. AB - OBJECTIVES: Although preoperative preparation programs were once common, most children currently undergoing outpatient surgery are first exposed to the hospital on the day of the procedure. It is advocated that these outpatient children undergo the preparation just prior to surgery. AIM: To assess the amount of time that healthcare providers spend with children and families on the day of surgery in the preoperative area. MATERIALS AND METHODS: The study used video infrastructure in the preoperative holding area of Yale New Haven Children's Hospital to record all interactions between children, families, and healthcare providers. Videotapes were coded to characterize and quantify behaviors of healthcare professionals. RESULTS: On the day of surgery, healthcare providers spent medians of 2.75-4.81 min interacting with children and parents in the preoperative area. Families spent a median of 46.5 min in the preoperative area. Healthcare professionals spent the most time in medical talk (averages of 42.5 48.2% of time spent with family) and little time was spent in nonmedical talk (range of 6.2-6.9% of time spent with family). Anesthesiologists and surgeons spent 28% and 18% of the interview in talk to children; admitting nurses spent more of the interview talking to children (43%). CONCLUSIONS: Families interact with healthcare providers for only a small proportion of the time they spent in the preoperative area. This is likely to be a result of increased production pressure in the perioperative settings and has implications for providing preparation for surgery on the morning of the procedure. PMID- 19187046 TI - Survey of pediatric continuing professional development undertaken by consultant anesthetists. AB - BACKGROUND: Whilst not all anesthetists have a regular pediatric commitment there is a need for out of hours cover of pediatric anesthesia. We have attempted to determine who covers pediatric anesthetic services in the District General Hospital setting. METHODS: A postal survey of 170 consultant anesthetists in nine District General Hospitals was conducted looking at who is responsible for pediatric anesthetics and emergencies out of hours as well as pediatric anesthetic experience, resuscitation training and continuing professional development (CPD). RESULTS: There was a 62% response rate with 98% of the consultants with on call duties also covering pediatric anesthetic emergencies. Fifty percent of consultants who responded were within 4 years of a pediatric specific resuscitation course, of which 93% had found it useful. However, 40% had never completed a pediatric resuscitation course or it could be considered out of date. Sixty-three percent of consultants had had some sort of pediatric anesthetic update in the last 4 years. CONCLUSIONS: General anesthetists are responsible for elective and emergency anesthetics as well as the care of critically ill children outside of specialist centers. This is despite a proportion of these consultants not having regular pediatric experience, not having completed a recent pediatric resuscitation course and without pediatric anesthetic CPD. PMID- 19187047 TI - Embracing the information age. PMID- 19187049 TI - Cultural diversity: the intention of nursing. AB - Nursing in the United States has expressed its intention of being a professional discipline that is culturally diverse. However, after examining the progress in this area, it is evident that nursing's movement toward cultural diversity has been slow and episodic. This article addresses cultural diversity progress in nursing and explores behaviors and actions that could enhance the cultural diversity of nursing. PMID- 19187048 TI - Concept analysis: malpractice and modern-day nursing practice. AB - TOPIC: The concept of malpractice can mean different things depending upon the context in which the term is used. This can lead to confusion about the standard of care required for nurses engaged in modern-day nursing practice. PURPOSE: This paper examines the attributes and characteristics of the concept of malpractice using Walker and Avant's (2005) eight-step methodology. SOURCES OF INFORMATION: CINAHL, PubMed, and PsychINFO. CONCLUSIONS: Exposure to malpractice liability is an unfortunate consequence of modern-day nursing practice. An understanding of malpractice will assist nurses in identifying situations that may expose them to legal liability and hopefully lead to improved patient care. PMID- 19187050 TI - The impact of healthcare issues on the future of the nursing profession: the resulting increased influence of community-based and public health nursing. AB - Several key issues, such as the necessity for cost containment, role conflicts between healthcare professions, nursing shortages, and organizational difficulties of healthcare organizations, significantly influence current healthcare delivery. These circumstances, which constitute a compelling need for responsive and effective change, are examined in terms of their impact upon the nursing profession. A review of the referenced journals and textbooks reveals that national nursing efforts will shift from acute care hospital-oriented provisions to community-based public health orientations. This evolution will result in improved health outcomes, less need for tertiary treatment, and savings for hospitals and insurance companies. PMID- 19187051 TI - Concept analysis of adherence in the context of cardiovascular risk reduction. AB - TOPIC: Cardiovascular disease is the number one cause of death in the United States. Examination of the concept of adherence is essential to provide informed patient-centered care to prevent the development and progression of this largely preventable disease. OBJECTIVE: The purpose of this concept analysis is to explore and clarify the concept of adherence. This analysis will provide a framework for those seeking a better understanding of patient decision-making, reduction of relapse to unhealthy behaviors, and increased long-term adherence to cardiovascular health recommendations. Reviews of major theories related to health behavior are reviewed in the second part of the article and are linked to the concept of adherence. METHOD: Following a review of the literature, Morse's guidelines were utilized to present a concept analysis of adherence and the related terms concordance and compliance from the disciplines of nursing, health psychology, ethics, pharmacy, and medicine. RESULTS: Adherence is dependent on the collaborative relationship between patient and healthcare provider. Adherence is influenced by the meaning of health, heart disease, and sense of personal risk as well as socioeconomic status, decision support, motivation, and desire for change, self-efficacy, and sources of credible health information. Attributes of successful adherence include alignment of patient behavior and health recommendations, mastery of new health knowledge and behavior, continued collaborative relationships between the patient and healthcare provider, and ability to meet outcome targets. CONCLUSION: Adherence is defined as persistence in the practice and maintenance of desired health behaviors and is the result of active participation and agreement. Adherence is dependent on the development of a concordant relationship and its measurement should be specific utilizing the correct tools. Special attention should be paid to shared decision-making between patient and healthcare provider. PMID- 19187052 TI - Violence against female student nurses in the workplace. AB - Violence, harassment, and bullying in the workplace are not new phenomena. However, the growing epidemic of violence in the health sector workplace is raising great concern among workers, employers, and governmental agencies across Australia, Canada, the United Kingdom, and the United States. National and international literature reveals that the prevalence of violence experienced by graduate and undergraduate female nursing students in the college and workplace settings is largely unknown. Moreover, the prevalence of violence is now recognized as a major health priority by the World Health Organization, the International Council of Nurses, and Public Services International. Even so, the number of nursing personnel affected by this problem continues to rise. A modified self-report online survey was used to ascertain the level of violence experienced by nursing students in their clinical placements. One hundred percent of those surveyed had experienced some type of workplace violence and the perpetrators were most often other staff members followed closely by patients. The American Association of Colleges of Nursing Position Statement recommends that all faculty prepare nurses to recognize and prevent all forms of violence in the workplace. This research seeks to develop practical approaches to better understand and prevent this global public health issue. PMID- 19187053 TI - International reflections on knowledge and use of the mirror in nursing practice. AB - TOPIC: There are no guidelines on the use of the mirror with patients in nursing practice. Nurses have used their experiences, personal preferences, and best guess practice models when to offer a mirror. PURPOSE: This international study provides insight into the current knowledge and uses of the mirror in nursing practice and, as such, provides a small foothold toward understanding the mirror as a multidimensional nursing tool. SOURCE OF INFORMATION: Ten healthcare providers from eight countries (South Africa, Egypt, Japan, the Netherlands, Panama, Russia, Singapore, and the United Kingdom) took part in semistructured questionnaire conversations via email. Thematic analysis was performed on the data. CONCLUSIONS: The results suggest the use of the mirror has a place in shaping nursing practice and is currently being used in nursing practice. Mirrors appear to be lacking in hospital environments. There is no known research to guide the nurse in the appropriate type of mirror to have in a hospital or any other healthcare environment. PMID- 19187054 TI - Stories appreciating both sides of the generation gap: baby boomer and Generation X nurses working together. AB - With nurses from the baby boomer and Generation X providing the majority of bedside nursing care, multigenerational differences are present in the workplace. The key to improved job satisfaction is the development of understanding and talking through differences between nurses of these age groups. From the perspective of a Generation X nurse, this paper addresses the differences in work ethic and values between these age groups and shows how such differences affect satisfaction with professional nursing. Improved job fulfillment can increase nursing retention and lessen the effects of the nursing shortage. PMID- 19187055 TI - Said another way. Nurses' role in cancer control. AB - TOPIC: Nurses' role in cancer control. PURPOSE: Many cancers are preventable and curable, if detected early. As the largest group of healthcare professionals, nurses can play a significant role in the control of cancer. Therefore, the purpose of this discussion is to highlight ways nurses can contribute to the prevention and early detection of cancer. It is meant to stimulate nurses' own ideas about the specific role they can play in cancer control, regardless of practice setting. INFORMATION SOURCES: Existing literature, professional practice. CONCLUSION: Nurses have the ability to serve as change agents in strengthening cancer control within the systems they practice, regardless of the type of setting. Nurses can engage in a number of cancer control activities such as providing patient education on cancer risk, cancer prevention, and early detection, and in the development of policy in support of cancer control activities in practice. PMID- 19187056 TI - Said another way. The real cost of higher education. PMID- 19187058 TI - The human premotor oculomotor brainstem system - can it help to understand oculomotor symptoms in Huntington's disease? AB - Recent progress in oculomotor research has enabled new insights into the functional neuroanatomy of the human premotor oculomotor brainstem network. In the present review, we provide an overview of its functional neuroanatomy and summarize the broad range of oculomotor dysfunctions that may occur in Huntington's disease (HD) patients. Although some of these oculomotor symptoms point to an involvement of the premotor oculomotor brainstem network in HD, no systematic analysis of this functional system has yet been performed in brains of HD patients. Therefore, its exact contribution to oculomotor symptoms in HD remains unclear. A possible strategy to clarify this issue is the use of unconventional 100-microm-thick serial tissue sections stained for Nissl substance and lipofuscin pigment (Nissl-pigment stain according to Braak). This technique makes it possible to identify the known nuclei of the premotor oculomotor brainstem network and to study their possible involvement in the neurodegenerative process. Studies applying this morphological approach and using the current knowledge regarding the functional neuroanatomy of this human premotor oculomotor brainstem network will help to elucidate the anatomical basis of the large spectrum of oculomotor dysfunctions that are observed in HD patients. This knowledge may aid clinicians in the diagnosis and monitoring of the disease. PMID- 19187059 TI - A quantitative study of the pathological changes in the cerebellum in 15 cases of variant Creutzfeldt-Jakob disease (vCJD). AB - AIMS: To determine in the cerebellum in variant Creutzfeldt-Jakob disease (vCJD): (i) whether the pathology affected all laminae; (ii) the spatial topography of the pathology along the folia; (iii) spatial correlations between the pathological changes; and (iv) whether the pathology was similar to that of the common methionine/methionine Type 1 subtype of sporadic CJD. METHODS: Sequential cerebellar sections of 15 cases of vCJD were stained with haematoxylin and eosin, or immunolabelled with monoclonal antibody 12F10 against prion protein (PrP) and studied using spatial pattern analysis. RESULTS: Loss of Purkinje cells was evident compared with control cases. Densities of the vacuolation and the protease-resistant form of prion protein (PrP(Sc)) (diffuse and florid plaques) were greater in the granule cell layer (GL) than the molecular layer (ML). In the ML, vacuoles and PrP(Sc) plaques occurred in clusters regularly distributed along the folia with larger clusters of vacuoles and diffuse plaques in the GL. There was a negative spatial correlation between the vacuoles and the surviving Purkinje cells in the ML. There was a positive spatial correlation between the vacuoles and diffuse PrP(Sc) plaques in the ML and GL. CONCLUSIONS: (i) all laminae were affected by the pathology, the GL more severely than the ML; (ii) the pathology was topographically distributed along the folia especially in the Purkinje cell layer and ML; (iii) pathological spread may occur in relation to the loop of anatomical connections involving the cerebellum, thalamus, cerebral cortex and pons; and (iv) there were pathological differences compared with methionine/methionine Type 1 sporadic CJD. PMID- 19187060 TI - The microglial/macrophagic response at the tumour-brain border of invasive meningiomas. AB - AIMS: Little is known about the immune response of the brain to invasive meningiomas. The present study was based upon the hypothesis that the microglial/macrophagic response towards brain-invasive meningiomas is dependent on the intactness of the pial-glial basement membrane. METHODS: We immunostained sections from 40 brain-invasive meningiomas that were graded according to World Health Organization (WHO) 2007 criteria. Thirty-three tumours were histologically WHO grade II (18, 'otherwise benign', and 15, 'otherwise atypical'), and seven, grade III. Microglial/macrophagic cells were labelled with antibodies directed against major histocompatibility complex class II, CD68, CD14 and CD163. Anti collagen IV was used to visualize basement membranes. RESULTS: Twenty-five per cent (10/40) meningiomas (1/18 WHO grade II 'otherwise benign', 3/15 grade II 'otherwise atypical' and 6/7 WHO grade III) contained microglial/macrophagic cells at the tumour-brain border. The presence of these cells correlated with the absence of the pial-glial basement membrane (BM) and with WHO grade III. The monocytic response was of two kinds: one consisted of a dense layer of mononuclear cells at the tumour-brain border in nine cases, the other of an elevated number of microglial cells expressing CD14 or CD163 (two cases). CONCLUSIONS: The immune response at the tumour-brain interface correlates with the absence of the pial-glial BM and with malignancy grade. It remains to be established whether the mononuclear cells at the tumour-brain border are native microglia or blood-derived macrophages. PMID- 19187061 TI - Early transient presence of implanted bone marrow stem cells reduces lesion size after cerebral ischaemia in adult rats. AB - AIMS: Previous studies on the therapeutic time window for intravascular administration of bone marrow stem cells (BMSCs) after stroke have shown that early intervention (from 3 h after onset) in the middle cerebral artery occlusion (MCAO) rat model is the most effective approach to reduce ischaemic lesion size. We have confirmed these observations but noticed that 2 weeks after transplantation, almost none of the grafted BMSCs could be detected in or around the lesion. The present experiments aimed to assess the fate and kinetics of intravascularly injected BMSCs shortly after administration in correlation to the development of the ischaemic lesion after MCAO. METHODS: We administered a syngeneic suspension of complete (haematopoietic and mesenchymal) BMSCs via the carotid artery to rats at 2 h after MCAO onset. We examined the distribution and tissue location of BMSCs within the first 24 h after arterial administration by perfusion-fixating rats and performing immunohistochemical analysis at different time points. RESULTS: The vast majority (>95%) of BMSCs appeared to become trapped in the spleen shortly after injection. Six hours after implantation, together with the appearance of activated microglia, the first BMSCs could be detected in and around the lesion; their number gradually increased during the first 12 h after implantation but started to decrease at 24 h. The implanted BMSCs were surrounded by activated and phagocytotic microglia. CONCLUSION: Our results show that ischaemic lesion size reduction can already be achieved by the early transient presence at the lesion site of intravascularly implanted BMSCs, possibly mediated via activated microglia. PMID- 19187062 TI - Muscle histopathology in myasthenia gravis with antibodies against MuSK and AChR. AB - AIMS: We compared myopathological features in myasthenia gravis (MG) patients with antibodies against AChR (seropositive) and muscle-specific tyrosin-kinase (MuSK). While the immunopathogenesis of seropositive MG is well known, there is a lack of pathological studies in anti-MuSK antibody-positive (MuSK+) MG. METHODS: We analysed skeletal muscle biopsy features of 13 MG patients: 6 MuSK+ (all women) and 7 anti-AchR antibody-positive (AChR+) (2 women and 5 men). In our histopathological examination, we quantified the atrophy factor of both fibre types, and the extent of minicores, myofibrillar disarray, cytochrome c oxidase (COX)-negative fibres, mitochondrial aggregates and fibre type grouping. RESULTS: Mean muscle fibre atrophy factor was higher in AChR+ MG than MuSK+ MG, both in type I fibres (494 vs. 210) and particularly in type II fibres (1023 vs. 300). Fibre type grouping was observed in AChR+ MG whereas COX-negative fibres were common in MuSK+ MG. Bulbar muscles were more severely affected in MuSK+ MG and the disease was more severe: the onset was usually earlier (39 years) with Myasthenia Gravis Foundation of America score III in MuSK+ MG, and score II was found in AChR+ MG (62 years). CONCLUSIONS: Muscle biopsies of MuSK+ MG show myopathic signs with prominent mitochondrial abnormalities, whereas neurogenic features and atrophy are more frequently found in AChR+ MG. The mitochondrial impairment could explain the oculo-bulbar involvement in MuSK+ MG. PMID- 19187063 TI - Familial prion disease with a novel serine to isoleucine mutation at codon 132 of prion protein gene (PRNP). PMID- 19187065 TI - Alpha-synuclein pathology and Parkinsonism associated with POLG1 mutations and multiple mitochondrial DNA deletions. PMID- 19187064 TI - Cerebral amyloid angiopathy: micro-haemorrhages demonstrated by magnetic resonance susceptibility-weighted imaging. PMID- 19187066 TI - Evidence against a reliable staging system of alpha-synuclein pathology in Parkinson's disease. PMID- 19187068 TI - Metabolic syndrome, non-alcoholic fatty liver disease and hepatitis C virus: impact on disease progression and treatment response. AB - Non-alcoholic fatty liver disease (NAFLD), a spectrum of liver disease ranging from simple steatosis to non-alcoholic steatohepatitis, is increasingly recognized as the hepatic manifestation of metabolic syndrome and is an important cause of liver-related morbidity and mortality. It is among the most common forms of liver disease. NAFLD reflects abnormal partitioning of fat, such that fat deposition is increased in the liver, and provides a link between NAFLD and the metabolic syndrome, a constellation of metabolic disorders that can also be associated with visceral fat or central adiposity. Together, the features of the metabolic syndrome presage overt diabetes and increase cardiovascular risk. Hepatitis C virus (HCV) appears to exacerbate the metabolic syndrome by eliciting increased insulin resistance (IR) and promoting truncal obesity. Moreover, the concomitant presence of HCV and NAFLD is associated with an increased likelihood of diabetes, hypertension and/or hypertriglyceridaemia. Metabolic abnormalities have been shown to influence response to treatment such that the presence of IR or obesity reduces the likelihood of a sustained virological response (SVR); conversely, SVR has been demonstrated to ameliorate IR and improve beta-cell function. Clinically, these data suggest that attention must be paid not only to optimizing antiviral response but also to screening for and treatment of the various components of the metabolic syndrome. PMID- 19187069 TI - Hepatitis C, insulin resistance and diabetes: clinical and pathogenic data. AB - Epidemiological data indicate a strong risk for development of insulin resistance (IR), and, ultimately, overt diabetes mellitus (DM) in patients with chronic hepatitis C virus (HCV) infection. Steatosis, or fatty liver, is closely linked with IR in persons without HCV, such as those with metabolic syndrome, primarily due to increased visceral fat leading to altered adipokine production and increased free fatty acid (FFA) release. Moreover, there is evidence that liver fat can have an impact on the development of hepatic IR independently of changes in adipose tissue. Multiple mechanisms can account for the development of IR in patients with chronic HCV. In particular, there is evidence for a triangular interaction between steatosis, inflammatory processes and IR. In patients infected by the genotype 1 virus, steatosis is strongly related to IR, leading to a metabolic steatosis, while, in genotype 3 patients, steatosis is related to viral load in the context of a viral steatosis. Chronic inflammatory processes in the liver may be mediated by persistently activated macrophages and other immune cells, with concomitant overproduction of pro-inflammatory cytokines such as tumour necrosis factor-alpha. Activation of inflammatory pathways, together with increased levels of FFAs, can disrupt hepatocyte intracellular pathways and inhibit insulin signalling, leading to IR. Molecular studies have also shown that the HCV core protein can directly inhibit the insulin signalling pathway and increase reactive oxygen species production, both of which can further exacerbate IR. The available data provide an understanding of chronic HCV whereby chronic inflammatory processes, steatosis and IR contribute to each other, leading to an increased risk of DM, and its associated poor outcomes, in persons with chronic HCV. PMID- 19187070 TI - Hepatitis C virus, steatosis and lipid abnormalities: clinical and pathogenic data. AB - Abnormal accumulation of fat in the liver (steatosis) is commonly observed in hepatitis C virus (HCV) infection, and the severity of steatosis has been well correlated with the degree of hepatic fibrosis. In patients with chronic HCV infection, steatosis may occur in conjunction with other metabolic risk factors such as insulin resistance and the metabolic syndrome. This was observed primarily in patients infected with non-genotype 3 virus. Otherwise, in HCV infected patients, especially those infected with genotype 3a, reductions in total cholesterol as well as high-density lipoprotein and low-density lipoprotein cholesterol are observed compared with matched controls, and the normalization of these parameters appears to be an important correlate of the response to antiviral therapy. In that setting, the pathogenic mechanisms involved in HCV induced steatosis are mediated in large part by the HCV core protein, whose expression is associated with lipid droplet accumulation, changes in lipogenic gene expression and/or the activity of lipogenic proteins, and effects on mitochondrial oxidative function. The importance of genes such as peroxisome proliferator-activated receptor-alpha and the proteasome activator PA28-gamma in HCV-mediated steatosis has been elucidated from studies in genetically altered mice, and the manipulation of these and other pathways may provide an avenue for therapeutic intervention. PMID- 19187072 TI - Metabolic abnormalities in chronic hepatitis C. Introduction. PMID- 19187071 TI - Hepatitis C, human immunodeficiency virus and metabolic syndrome: interactions. AB - Significant concerns have been raised about the metabolic effects of antiretroviral medication, including the classic triad of dyslipidaemia, insulin resistance (IR) and characteristic alterations in fat distribution (lipoatrophy and lipohypertrophy). Co-infection with hepatitis C appears to exacerbate IR, reduce serum lipids and induce prothrombotic changes in the treated human immunodeficiency virus patient. The effects of co-infection are complex. While combination antiretroviral therapy has been shown to be associated with an increased risk of cardiovascular events through promotion of dyslipidaemia, IR and fat redistribution, co-infection exacerbates IR while reducing serum lipids. Co-infection also promotes a prothrombotic state characterized by endothelial dysfunction and platelet activation, which may enhance risk for cardiovascular disease. Consideration must be given to selection of appropriate treatment regimens and timing of therapy in co-infected patients to minimize metabolic derangements and, ultimately, reduce cardiovascular risk. PMID- 19187073 TI - von Willebrand factor is a major determinant of ADAMTS-13 decrease during mouse sepsis induced by cecum ligation and puncture. AB - SUMMARY BACKGROUND: During sepsis, von Willebrand factor (VWF) is abundantly secreted; the main mechanism regulating its size involves specific proteolysis by the metalloprotease ADAMTS-13. OBJECTIVES: To determine whether ADAMTS-13 consumption due to its binding to, and/or cleavage, of VWF contributes to its decrease during sepsis and whether abrogating or enhancing ADAMTS-13 activity influences sepsis outcome. METHODS: ADAMTS-13 activity was evaluated in a model of sepsis induced by cecum ligature and puncture (CLP) in wild-type and Vwf(-/-) mice. Sepsis outcome was studied in those mice and in Adamts-13(-/-) mice. Finally, survival was studied in wild-type mice injected hydrodynamically with the human ADAMTS-13 gene. RESULTS: In wild-type mice, CLP-induced sepsis elicited a significant ADAMTS-13 decrease, and a strong negative correlation existed between VWF and ADAMTS-13. In Vwf(-/-) mice, CLP also induced severe sepsis, but ADAMTS-13 was not significantly diminished. Notably, Vwf(-/-) mice lived significantly longer than wild-type mice. In contrast, Adamts-13(-/-) mice and wild-type mice were comparable with regard to thrombocytopenia, VWF concentrations, absence of thrombi, and survival. Hydrodynamic hADAMTS-13 gene transfer with the pLIVE expression vector resulted in high and stable ADAMTS13 activity in CLP mice; however, no impact on survival was observed. CONCLUSIONS: VWF secretion is a major determinant of ADAMTS-13 decrease in the CLP model, and plays an important role in sepsis-induced mortality, but the complete absence of its regulating protease, ADAMTS-13, had no detectable impact in this sepsis model. Furthermore, increasing ADAMTS-13 activity had no impact on survival. PMID- 19187074 TI - JAK2V617F mutation in patients with arterial thrombosis in the absence of overt myeloproliferative disease. PMID- 19187076 TI - AVE5026, a new hemisynthetic ultra-low-molecular-weight heparin for the prevention of venous thromboembolism in patients after total knee replacement surgery--TREK: a dose-ranging study. AB - BACKGROUND: AVE5026 is a new hemisynthetic ultra-low-molecular-weight heparin, with a novel anti-thrombotic profile resulting from high anti-factor (F)Xa activity and residual anti-FIIa activity. AVE5026 is in clinical development for venous thromboembolism (VTE) prevention, a frequent complication after total knee replacement (TKR) surgery. OBJECTIVES: This study evaluated the dose-response of AVE5026 for the prevention of VTE in patients undergoing TKR surgery. PATIENTS/METHODS: In this parallel-group, double-blind, double-dummy study, 690 patients were randomized, and 678 treated with once-daily doses of AVE5026 (5, 10, 20, 40, or 60 mg) or enoxaparin 40 mg in the calibrator arm. The primary efficacy end point was VTE until post-operative day 11, defined as deep vein thrombosis (DVT) detected by bilateral venography, symptomatic DVT, non-fatal pulmonary embolism (PE) and VTE-related death. The primary safety outcome was the incidence of major bleeding. RESULTS: The primary efficacy outcome was assessed in 464 patients. There was a significant dose-response across the five AVE5026 groups for VTE prevention (P<0.0001), with the incidence of VTE ranging from 5.3% to 44.1% compared with 35.8% in the enoxaparin group and for proximal DVT (P=0.0002). Also, a significant dose-response for AVE5026 was seen for major bleeding (P=0.0231) and any bleeding (P=0.0003). Six patients in the AVE5026 groups, four in the 60 mg group, experienced major bleeding; none did in the enoxaparin group. CONCLUSIONS: The safety and efficacy results of this study suggest that a AVE5026 dose of between 20 and 40 mg presents an adequate benefit to-risk ratio. PMID- 19187075 TI - Lipid biomarkers, hormone therapy and the risk of venous thromboembolism in women. AB - BACKGROUND: Published reports of a relationship between lipids and incident venous thromboembolism (VTE) are conflicting. OBJECTIVES: To clarify the relationship between lipids and VTE risk in healthy women, including potential effect modification by hormone therapy (HT). PATIENTS/METHODS: Among 27 081 initially healthy women followed prospectively for incident VTE, we measured a full panel of lipid biomarkers, including total cholesterol, low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), triglycerides and apolipoproteins A-I (apo A-I) and B(100). RESULTS: During a median follow-up of 11.4 years, VTE occurred in 355 women. We observed no relationship between any of the lipids and VTE risk. However, when unprovoked VTE was considered separately (n=161), both HDL-C and apo A-I were positively associated with risk. Fully adjusted hazard ratios (HR) and 95% confidence intervals (CI) for extreme tertiles of HDL-C and apo A-I were 1.75 (1.13-2.73) and 1.70 (1.10-2.62), respectively. After stratifying by HT use, this relationship was present only among HT users; the HRs for unprovoked VTE for extreme tertiles of HDL-C and apo A-I were 3.58 (1.69-7.58) and 2.88 (1.29-6.42) among users, but only 0.79 (0.39-1.62) and 0.89 (0.50-1.57) among non-users. The interactions were statistically significant (each Pinteraction<0.05). CONCLUSIONS: We observed little evidence that lipid levels predict risk of incident VTE among non-users of HT. High levels of HDL-C and apo A-I associate with unprovoked VTE risk among HT users. This observation likely reflects prothrombotic effects of HT that are concomitant with HDL-C and apo A-I levels, rather than direct effects of those lipids. PMID- 19187077 TI - Chemotherapeutic agents doxorubicin and epirubicin induce a procoagulant phenotype on endothelial cells and blood monocytes. AB - BACKGROUND: Although chemotherapy is associated with an increased risk of thrombosis, the pathogenic mechanisms by which chemotherapeutic agents exert prothrombotic effects are unclear. OBJECTIVES: In this study we explored the possibility that chemotherapeutic agents doxorubicin, epirubicin, 5-fluorouracil and methotrexate induce a procoagulant phenotype on vascular endothelial cells and/or on blood monocytes. METHODS: Thrombin generation was measured in defibrinated plasma exposed to chemotherapy-treated human umbilical vein endothelial cells (HUVECs). Tissue factor activity assays were performed on chemotherapy-treated HUVECs and blood monocytes. The effects of chemotherapy drugs on phosphatidylserine exposure and the protein C pathway were also measured. RESULTS: Exposure of defibrinated plasma to either doxorubicin- or epirubicin-treated HUVECs resulted in an increase in plasma thrombin generation. The procoagulant activity of doxorubicin- and epirubicin-treated HUVECs reflects an increase in tissue factor activity and phosphatidylserine exposure. Doxorubicin-mediated increase in tissue factor activity is related to increased levels of phosphatidylserine rather than to protein disulfide isomerase activity, and is likely to involve reactive oxygen species generation. Unlike doxorubicin, epirubicin does not have an impact on the protein C anticoagulant pathway. Interestingly, neither methotrextate nor 5-fluorouracil altered endothelial or monocyte hemostatic properties. CONCLUSIONS: These studies suggest that doxorubicin and epirubicin have the greatest 'prothrombotic potential' by virtue of their ability to alter endothelial and monocyte hemostatic properties. PMID- 19187078 TI - Non-classical anti-factor VIII C2 domain antibodies are pathogenic in a murine in vivo bleeding model. AB - OBJECTIVE: The pathogenicity of anti-human factor (F) VIII monoclonal antibodies (MAbs) was tested in a murine bleeding model. METHODS: MAbs were injected into the tail veins of hemophilia A mice to a peak plasma concentration of 60 nm, followed by injection of human B domain-deleted FVIII at 180 U kg(-1), producing peak plasma concentrations of approximately 2 nm. At 2 h, blood loss following a 4-mm tail snip was measured. The following MAbs were tested: (i) 4A4, a type I anti-A2 FVIII inhibitor, (ii) I54 and 1B5, classical type I anti-C2 inhibitors, (iii) 2-77 and B45, non-classical type II anti-C2 inhibitors, and (iv) 2-117, a non-classical anti-C2 MAb with inhibitory activity less than 0.4 Bethesda Units per mg IgG. RESULTS: All MAbs except 2-117 produced similar amounts of blood loss that were significantly greater than control mice injected with FVIII alone. Increasing the dose of FVIII to 360 U kg(-1) overcame the bleeding diathesis produced by the type II MAbs 2-77 and B45, but not the type I antibodies, 4A4, I54, and 1B5. These results were consistent with the in vitro Bethesda assay in which 4A4 completely inhibited both 1 U mL(-1) and 3 U mL(-1) FVIII, while there was 40% residual activity at saturating concentrations of 2-77 at either concentration of FVIII. CONCLUSIONS: For patients with an inhibitor response dominated by non-classical anti-C2 antibodies both the in vivo and in vitro results suggest that treatment with high-dose FVIII rather than bypassing agents may be warranted. PMID- 19187079 TI - The pharmacokinetics of idraparinux, a long-acting indirect factor Xa inhibitor: population pharmacokinetic analysis from Phase III clinical trials. AB - BACKGROUND: Idraparinux, a long-acting synthetic pentasaccharide, is a specific antithrombin-dependent inhibitor of activated factor X that has been investigated in the treatment and prevention of thromboembolic events. OBJECTIVES: To characterize the population pharmacokinetic profile of idraparinux in patients enrolled in van Gogh and Amadeus Phase III clinical trials. PATIENTS AND METHODS: Idraparinux was administered once-weekly subcutaneously at a dose of 2.5 mg, or 2.5 mg (first dose) and then 1.5 mg for patients with severe renal insufficiency (creatinine clearance<30 mL min(-1)). A population pharmacokinetic model was developed using data from 704 patients with acute deep-vein thrombosis or pulmonary embolism, 1310 patients suffering from atrial fibrillation, and 40 healthy subjects. Potential covariates analyzed included demographics (age, sex, weight and ethnicity), and serum creatinine and creatinine clearance determinations. RESULTS: A three-compartment model best described idraparinux pharmacokinetics, with interindividual variability on clearance, central volume of distribution, and absorption rate constant; residual variability was low. Typical clearance, central volume of distribution, absorption rate constant and volume of distribution at steady-state were 0.0255 L h(-1), 3.36 L, 1.37 h and 30.8 L, respectively. Peak concentration was reached at 2.5 h. The terminal half life was 66.3 days and time to steady-state was 35 weeks. At steady-state, exposures were similar for patients without and with severe renal impairment receiving adjusted-dose. Creatinine clearance was the most important covariate affecting idraparinux clearance. The particular characteristics of idraparinux- rapid onset of action and long-acting anticoagulant effect--offer interesting clinical perspectives currently under investigation with idrabiotaparinux, the reversible biotinylated form of idraparinux. PMID- 19187080 TI - Atherosclerosis in thrombotic primary antiphospholipid syndrome. AB - BACKGROUND: Primary antiphospholipid syndrome (PAPS) is characterized by arterial and venous thrombosis, pregnancy loss, often recurrent, in the presence and persistence on antiphospholipid antibodies (aPL). The issue of early atherosclerosis, as evaluated by measuring carotid intima media thickness (IMT), associated with aPL, has been limitedly explored in PAPS. METHODS: In an age- and sex-matched case-double-control study, intima media thickeness of carotid arteries was measured using high-resolution B-mode ultrasound in 49 thrombotic PAPS patients (18 M, 31 F, mean age 37+/-11), in 49 patients who suffered thrombosis for inherited thrombophilia and 49 healthy subjects. RESULTS: Average carotid IMT was always greater in PAPS than control patients (common carotid P=0.004, bifurcation P=0.013, internal carotid P=0.001). By dividing participants into age tertiles most of the difference was explained by greater IMT of PAPS patients in the second (common carotid P=0.003, bifurcation P=0.023, internal carotid P=0.003) and third tertiles (common carotid P=0.03, bifurcation P=0.004, internal carotid P=0.007). CONCLUSIONS: Premature atherosclerosis is a clinical feature of our thrombotic PAPS patients. PMID- 19187081 TI - Racial differences in endotoxin-induced tissue factor-triggered coagulation. AB - BACKGROUND: Racial differences in coagulation are poorly understood. While some studies suggest a 'prothrombotic' coagulation profile in blacks compared with whites, others report an increased bleeding risk for blacks in various clinical settings. Moreover, preclinical data suggest a link between the Duffy antigen (=DARC, Duffy antigen receptor of chemokines) and coagulation. OBJECTIVES: Based on our previous research in Duffy antigen negative Africans, we hypothesized that Africans have an attenuated procoagulant response compared with Caucasians in a model of lipopolysaccharide (LPS)-induced, tissue factor (TF)-triggered coagulation activation. PATIENTS/METHODS: Healthy male volunteers (16 Duffy negative Africans, 16 Duffy-positive Caucasians) received 2 ng kg(-1) LPS, and outcome parameters were measured using enzyme immunoassays and real-time polymerase chain reaction (RT-PCR, Taqman). RESULTS: LPS increased microparticle (MP)-associated TF procoagulant activity (PCA) less in Africans than Caucasians. Africans had reduced in vivo thrombin formation compared with Caucasians: they generated less thrombin-antithrombin (TAT) complexes (10.4 pg mL(-1) vs. 23.0 pg mL(-1), P<0.0001) and less prothrombin fragments (F1+2) (337 pmol mL(-1) vs. 819 pmol mL(-1), P<0.0001). Consistently, Africans also had decreased fibrin formation (D-dimer: 0.3 pg mL(-1) vs. 0.5 pg mL(-1), P=0.02). CONCLUSION: Duffy negative subjects of African descent have a markedly reduced procoagulant response in a model of LPS-induced, TF-triggered coagulation activation compared with Duffy-positive healthy Caucasians. PMID- 19187082 TI - School injury among Ottawa-area children: a population-based study. AB - BACKGROUND: Injuries are the leading cause of death among Canadian children and are responsible for a substantial proportion of hospitalizations and emergency department visits. This investigation sought to identify the factors associated with the likelihood of sustaining an injury at school among Ottawa-area children. METHODS: Children presenting to Ottawa-area hospitals and urgent care clinics from January to December 2002 (n = 24,074) were included for analysis. The frequency of school injuries by sex, age group, type of injury, and hospitalization was analyzed. Multivariate logistic regression was used to assess the factors associated with sustaining an injury at school. The school activities most associated with injury and the most frequent types of school injuries were assessed. RESULTS: A total of 4287 Ottawa-area children were injured at school in 2002, representing 18% of all injuries. Children aged 5-9 years and 10-14 years were more likely to have school injuries than older children (aged 15-19 years) (OR = 3.07, 95% CI = 2.77-3.40 and OR = 3.10, 95% CI = 2.83-3.37, respectively). The most frequently encountered school injuries were fractures (n = 1132) and musculoskeletal injury (n = 907). The most frequent mechanisms of school injuries were "playing" (n = 1004) and "informal sports" (n = 1503). CONCLUSIONS: Many children get hurt at school, particularly during informal recreation activities. Environmental modification and increased supervision are strategies that may reduce school injuries. PMID- 19187083 TI - Measuring children's self-efficacy and proxy efficacy related to fruit and vegetable consumption. AB - BACKGROUND: Social cognitive theory describes self-efficacy and proxy efficacy as influences on fruit and vegetable consumption (FVC). Proxy efficacy was defined as a child's confidence in his or her skills and abilities to get others to act in one's interests to provide fruit and vegetable (FV) opportunities. The purpose of this study was to develop a scale assessing children's self-efficacy and proxy efficacy for FVC at after-school programs and at home. METHODS: Elementary-aged children (n = 184) attending 7 after-school programs completed a self-efficacy questionnaire relevant to FVC. Questionnaire validity was investigated with exploratory factor analysis and mixed-model analysis of covariance. Internal consistency reliability and readability were also assessed. RESULTS: The questionnaire assessed 4 constructs: self-efficacy expectations for fruit consumption, self-efficacy expectations for vegetable consumption, proxy efficacy to influence parents to make FV available, and proxy efficacy to influence after school staff to make FV available. Children perceiving FV opportunities in after school had greater self-efficacy expectations for FVC and greater proxy efficacy to influence after-school staff compared to students who did not perceive FV opportunities. Children attending schools of higher socioeconomic status (SES) and less diversity were more confident they could influence their parents to make FV available than students attending lower SES and less diverse schools. Adequate internal consistency and test-retest reliabilities were established. CONCLUSIONS: Self-efficacy is a multicomponent construct that can be assessed in children using the reliable and valid instrument evaluated by the current study. PMID- 19187084 TI - Alcohol use among rural middle school students: adolescents, parents, teachers, and community leaders' perceptions. AB - BACKGROUND: Although rural adolescents use of alcohol is at some of the highest rates nationally, rural adolescent alcohol use has not been studied extensively. This study examines how community attitudes and behaviors are related to adolescent drinking in rural environments. METHODS: Data were gathered in 22 rural communities in the Upper Midwest (North Dakota, South Dakota, Wisconsin, and Wyoming). Surveys were collected from 1424 rural sixth- to eighth-grade adolescents and 790 adults, including parents, teachers, and community leaders. Census data were also collected. RESULTS: Drinkers differed from nondrinkers by the following factors: higher perceptions of peer, parental, and overall community drinking, as well as lower levels of parental closeness and religiosity. Factors distinguishing binge and nonbinge drinkers were increased drinking to reduce stress, drinking to fit in, perceptions of peer drinking, and perceived lack of alternatives to drinking. Parents were significantly less likely to perceive adolescent alcohol use as a problem than other community adults; school officials were most likely to perceive it as a problem. Parental perceptions were also the least correlated to actual adolescent use, while adolescent perceptions were the most highly correlated. CONCLUSIONS: Community fac tors such as overall prevalence of drinking, community support, and controls against drinking are important predictors of reported use in early adolescence. School officials were more likely to view adolescent alcohol use as a problem than were parents. School officials' perceptions of adolescent use were also more related to actual adolescent use than were parental perceptions of adolescent use. PMID- 19187085 TI - A study of smoking disparity and factors associated with children smoking behavior in the mountain and city schools in Taiwan. AB - BACKGROUND: While higher smoking prevalences have been better described for adults and adolescents in the mountainous areas than in the plain area in Taiwan, no studies have previously examined whether this disparity begins with children in elementary schools. The purpose of this study was thus designed to explore clustering in smoking behavior among elementary school children attending mountain schools compared to those attending city schools. METHODS: This study analyzed data obtained by a survey on smoking behavior collected during the School Smoking Survey Project performed in 13 elementary schools of Taoyuan County, Taiwan. Overall, 1585 third and fourth graders (mean age 8.9 years) participated in the study. A multilevel logistic regression analysis was performed to explore the effects of school location on individual smoking behavior among elementary school children while controlling for individual-level characteristics. RESULTS: Overall, 34.9% of the elementary school students in the mountain schools reported having tried cigarette smoking compared to only 9.6% of students from city schools. Students attending mountain schools had a greater likelihood of reporting smoking than students attending city schools after controlling for individual-level characteristics (OR = 2.57, 95% CI = 1.10-5.99). CONCLUSIONS: A significant individual clustering in smoking behavior was found among third- and fourth-grade children attending mountain schools. The new findings suggest that the adult geographic smoking disparity begins in elementary school. Interventions aimed at reducing smoking disparity in adults need to target elementary schools in high-risk locations. PMID- 19187086 TI - Racial, ethnic, and sex differences in the associations between violence and self reported health among US high school students. AB - BACKGROUND: Involvement in interpersonal violence or suicidal behaviors can have a significant impact on an adolescent's physical health. Similarly, previous research has suggested that lived experiences, more than the presence or absence of physical ailments, can significantly influence self-assessed health status among adolescents. The purpose of this study was to examine the cross-sectional associations between involvement in violence and poor or fair self-reported health among US high school students. METHODS: Data were obtained from the 2005 national Youth Risk Behavior Survey (n = 13,953). Logistic regression analyses were conducted to determine the associations between violence-related measures and self-reported health while controlling for demographic characteristics and potential confounders. Analyses are presented for students overall and stratified by sex and race/ethnicity. RESULTS: Overall, 7.2% of students reported fair or poor self-rated health. Having been in a physical fight, having been injured in a physical fight, having attempted suicide, and having not gone to school because of safety concerns were significantly associated with fair or poor self-rated health after controlling for demographic characteristics and other potential confounders. Differences associated with race/ethnicity and sex are identified. CONCLUSIONS: Four of the 5 violence-related measures included in these analyses were significantly associated with fair or poor self-rated health. Future studies should consider the impact of involvement in violent behaviors and perceptions of both physical and mental well-being. PMID- 19187087 TI - Emotional/Behavioral difficulties and mental health service contacts of students in special education for non-mental health problems. AB - BACKGROUND: Emotional/behavioral difficulties and mental health (MH) service contacts of 3 groups of youth were compared: students in special education for non-MH problems, students in special education for MH problems, and youth not in special education. METHODS: Parents reported the characteristics, special education placement, emotional/behavioral difficulties, and MH service contacts of 25,122 youth aged 6-17 years in the National Health Interview Survey. RESULTS: Two thirds of students in special education received special education services for non-MH problems. Among students in special education for non-MH problems, 17% had serious emotional/behavioral difficulties compared with 51% of students in special education for MH problems and 4% of youth not in special education. MH service contacts were examined only for youth whose difficulties significantly interfered with their ability to function in or out of school. Among youth with serious difficulties, the percentage of youth without a recent MH service contact was greater for students in special education for non-MH problems (40%) and youth not in special education (47%) compared with students in special education for MH problems (13%). CONCLUSIONS: Compared with youth not in special education, students in special education for non-MH problems were 4 times more likely to have serious emotional/behavioral difficulties but were just as likely as youth not in special education to lack a recent MH service contact. Study findings provide a national context for considering the MH screening/evaluation needs of students receiving special education for non-MH problems. PMID- 19187090 TI - Communication between neurons and astrocytes: relevance to the modulation of synaptic and network activity. AB - Neuromodulation is a fundamental process in the brain that regulates synaptic transmission, neuronal network activity and behavior. Emerging evidence demonstrates that astrocytes, a major population of glial cells in the brain, play previously unrecognized functions in neuronal modulation. Astrocytes can detect the level of neuronal activity and release chemical transmitters to influence neuronal function. For example, recent findings show that astrocytes play crucial roles in the control of Hebbian plasticity, the regulation of neuronal excitability and the induction of homeostatic plasticity. This review discusses the importance of astrocyte-to-neuron signaling in different aspects of neuronal function from the activity of single synapses to that of neuronal networks. PMID- 19187091 TI - D2 receptor-mediated inhibition of dopamine release in the rat striatum in vitro is modulated by CB1 receptors: studies using fast cyclic voltammetry. AB - Cannabinoid CB(1) receptors are highly expressed in the striatum where they are known to be co-localized with dopamine D(2) receptors. There is now strong evidence that cannabinoids modulate dopamine release in the brain. Using fast cyclic voltammetry, single pulse stimulation (0.1 ms; 10 V) was applied every 5 min and peak dopamine release was measured with a carbon fibre microelectrode. Application of the D(2) receptor agonist, quinpirole, inhibited single pulse dopamine overflow in a concentration-dependent manner (IC(50): 3.25 x 10(-8) M). The CB(1) receptor agonist WIN55212-2 (WIN; 1 microM) had no effect on single pulse dopamine release (93.9 +/- 6.6% at 60 min, n = 5) but attenuated the inhibitory effect of quinpirole (30 nM; quinpirole 39.0 +/- 4.2% vs. quinpirole + WIN, 48.2 +/- 3.7%, n = 5, p < 0.05). This affect was antagonized by the CB(1) receptor antagonist [N-(Piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4 methyl-1H-pyrazole-3-carboxamide] (AM-251, 1 microM). Dopamine release evoked by four pulses delivered at 1 Hz (4P1Hz) and 10 pulses delivered at 5 Hz (10P5Hz) was significantly inhibited by WIN [72.3 +/- 7.9% control (peak 4 to 1 ratio measurement) and 66.9 +/- 3.8% control (area under the curve measurement), respectively, p < 0.05; n = 6 for both]. Prior perfusion of WIN significantly attenuated the effects of quinpirole on multiple pulse-evoked dopamine release (4P1Hz: quinpirole, 28.4 +/- 4.8% vs. WIN + quinpirole, 52.3 +/- 1.2%; 10P5Hz: quinpirole, 29.5 +/- 1.3% vs. WIN + quinpirole, 59.4 +/-7.1%; p < 0.05 for both; n = 6). These effects were also antagonized by AM-251 (1 microM). This is the first report demonstrating a functional, antagonistic interaction between CB(1) receptors and D(2) autoreceptors in regulating rat striatal dopamine release. PMID- 19187092 TI - L-DOPA activates ERK signaling and phosphorylates histone H3 in the striatonigral medium spiny neurons of hemiparkinsonian mice. AB - In the dopamine-depleted striatum, extracellular signal-regulated kinase (ERK) signaling is implicated in the development of L-DOPA-induced dyskinesia. To gain insights on its role in this disorder, we examined the effects of L-DOPA on the state of phosphorylation of ERK and downstream target proteins in striatopallidal and striatonigral medium spiny neurons (MSNs). For this purpose, we employed mice expressing enhanced green fluorescent protein (EGFP) under the control of the promoters for the dopamine D(2) receptor (Drd2-EGFP mice) or the dopamine D(1) receptor (Drd1a-EGFP mice), which are expressed in striatopallidal and striatonigral MSNs, respectively. In 6-hydroxydopamine-lesioned Drd2-EGFP mice, L DOPA increased the phosphorylation of ERK, mitogen- and stress-activated kinase 1 and histone H3, selectively in EGFP-negative MSNs. Conversely, a complete co localization between EGFP and these phosphoproteins was observed in Drd1a-EGFP mice. The effect of L-DOPA was prevented by blockade of dopamine D(1) receptors. The same pattern of activation of ERK signaling was observed in dyskinetic mice, after repeated administration of L-DOPA. Our results demonstrate that in the dopamine-depleted striatum, L-DOPA activates ERK signaling specifically in striatonigral MSNs. This regulation may result in ERK-dependent changes in striatal plasticity leading to dyskinesia. PMID- 19187093 TI - The proteome of the presynaptic active zone: from docked synaptic vesicles to adhesion molecules and maxi-channels. AB - The presynaptic proteome controls neurotransmitter release and the short and long term structural and functional dynamics of the nerve terminal. Using a monoclonal antibody against synaptic vesicle protein 2 we immunopurified a presynaptic compartment containing the active zone with synaptic vesicles docked to the presynaptic plasma membrane as well as elements of the presynaptic cytomatrix. Individual protein bands separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis were subjected to nanoscale-liquid chromatography electrospray ionization-tandem mass spectrometry. Combining this method with 2-dimensional benzyldimethyl-n-hexadecylammonium chloride/sodium dodecyl sulfate-polyacrylamide gel electrophoresis and matrix-assisted laser desorption ionization time of flight and immunodetection we identified 240 proteins comprising synaptic vesicle proteins, components of the presynaptic fusion and retrieval machinery, proteins involved in intracellular signal transduction, a large variety of adhesion molecules and proteins potentially involved in regulating the functional and structural dynamics of the pre-synapse. Four maxi-channels, three isoforms of voltage-dependent anion channels and the tweety homolog 1 were co-isolated with the docked synaptic vesicles. As revealed by in situ hybridization, tweety homolog 1 reveals a distinct expression pattern in the rodent brain. Our results add novel information to the proteome of the presynaptic active zone and suggest that in particular proteins potentially involved in the short and long term structural modulation of the mature presynaptic compartment deserve further detailed analysis. PMID- 19187094 TI - Phosphodiesterase 5 inhibitors prevent 3,4-methylenedioxymethamphetamine-induced 5-HT deficits in the rat. AB - Phosphodiesterase 5 (PDE5) inhibitors are often used in combination with club drugs such as 3,4-methylenedioxymethamphetamine (MDMA or ecstasy). We investigated the consequences of such combination in the serotonergic system of the rat. Oral administration of sildenafil citrate (1.5 or 8 mg/kg) increased brain cGMP levels and protected in a dose-dependent manner against 5 hydroxytryptamine depletions caused by MDMA (3 x 5 mg/kg, i.p., every 2 h) in the striatum, frontal cortex and hippocampus without altering the acute hyperthermic response to MDMA. Intrastriatal administration of the protein kinase G (PKG) inhibitor, KT5823 [(9S, 10R, 12R)-2,3,9,10,11,12-Hexahydro-10-methoxy-2,9 dimethyl-1-oxo-9,12-epoxy-1H-diindolo[1,2,3-fg:3',2',1'-kl]pyrrolo[3,4 i][1,6]benzodiazocine-10-carboxylic acid, methyl ester)], suppressed sildenafil mediated protection. By contrast, the cell permeable cGMP analogue, 8 bromoguanosine cyclic 3',5'-monophosphate, mimicked sildenafil effects further suggesting the involvement of the PKG pathway in mediating sildenafil protection. Because mitochondrial ATP-sensitive K(+) channels are a target for PKG, we next administered the specific mitochondrial ATP-sensitive K(+) channel blocker, 5 hydroxydecanoic acid, 30 min before sildenafil. 5-hydroxydecanoic acid completely reversed the protection afforded by sildenafil, thereby implicating the involvement of mitochondrial ATP-sensitive K(+) channels. Sildenafil also increased Akt phosphorylation, and so the possible involvement of the Akt/endothelial nitric oxide synthase (eNOS)/sGC signalling pathway was analysed. Neither the phosphatidylinositol 3-kinase inhibitor, wortmannin, nor the selective eNOS inhibitor, L-N5-(1-iminoethyl)-L-ornithine dihydrochloride, reversed the protection afforded by sildenafil, suggesting that Akt/eNOS/sGC cascade does not participate in the protective mechanisms. Our data also show that the protective effect of sildenafil can be extended to vardenafil, another PDE5 inhibitor. In conclusion, sildenafil protects against MDMA-induced long-term reduction of indoles by a mechanism involving increased production of cGMP and subsequent activation of PKG and mitochondrial ATP-sensitive K(+) channel opening. PMID- 19187095 TI - Interleukin-6 is required for the early induction of glial fibrillary acidic protein in Schwann cells during Wallerian degeneration. AB - Signal transducer and activator of transcription 3 (STAT3) regulates gene transcription in response to cytokines and growth factors. In the central nervous system, STAT3 plays a role in neuroprotection and reactive gliosis after lesions. During peripheral nerve regeneration, a nerve injury-induced up-regulation of cytokines and growth factors accompanies STAT3 activation in sensory neurons and Schwann cells (SCs) even though its molecular details and functions are unknown. We then analyzed the ligands and functions of STAT3 activation in RT4 schwannoma cells and adult SCs in vitro and in vivo. We have identified that interleukin-6 (IL-6), but not ciliary neurotrophic factor, leukemia inhibitory factor, or ligands for receptor tyrosine kinases, activates STAT3 in SCs. The IL-6/STAT3 signaling in primary SCs and RT4 cells induced the gene expression of glial fibrillary acidic protein (GFAP), which is known to be required for the proper regeneration of the injured nerves. Finally, the GFAP induction in the sciatic nerves after injury was significantly delayed in IL-6-deficient mice. These findings indicate that IL-6 plays an important role in STAT3-dependent GFAP induction in SCs during peripheral nerve regeneration. PMID- 19187096 TI - Induction of chemokines, MCP-1, and KC in the mutant huntingtin expressing neuronal cells because of proteasomal dysfunction. AB - Huntington's disease is a hereditary neurodegenerative disorder caused by an aberrant polyglutamine expansion in the amino terminus of the huntingtin protein. The resultant mutant huntingtin form aggregates in neurons and causes neuronal dysfunction and degeneration in many ways including transcriptional dysregulation. Here, we report that the expression of mutant huntingtin in the mouse neuroblastoma cell results in massive transcriptional induction of several chemokines including monocyte chemoattractant protein-1 (MCP-1) and murine chemokine (KC). The mutant huntingtin expressing cells also exhibit proteasomal dysfunction and down-regulation of NF-kappaB activity in a time-dependent manner and both these phenomena regulate the expression of MCP-1 and KC. The expression of MCP-1 and KC are increased in the mutant huntingtin expressing cells in response to mild proteasome inhibition. However, the expression of MCP-1 and KC and proteasome activity are not altered and inflammation is rarely observed in the brain of 12-week-old Huntington's disease transgenic mice in comparison with their age-matched controls. Our result suggests that the mutant huntingtin induced proteasomal dysfunction can up-regulate the expression of MCP-1 and KC in the neuronal cells and therefore might trigger the inflammation process. PMID- 19187097 TI - PKG activity causes photoreceptor cell death in two retinitis pigmentosa models. AB - Photoreceptor degeneration in retinitis pigmentosa is one of the leading causes of hereditary blindness in the developed world. Although causative genetic mutations have been elucidated in many cases, the underlying neuronal degeneration mechanisms are still unknown. Here, we show that activation of cGMP dependent protein kinase (PKG) hallmarks photoreceptor degeneration in rd1 and rd2 human homologous mouse models. When induced in wild-type retinae, PKG activity was both necessary and sufficient to trigger cGMP-mediated photoreceptor cell death. Target-specific, pharmacological inhibition of PKG activity in both rd1 and rd2 retinae strongly reduced photoreceptor cell death in organotypic retinal explants. Likewise, inhibition of PKG in vivo, using three different application paradigms, resulted in robust photoreceptor protection in the rd1 retina. These findings suggest a pivotal role for PKG activity in cGMP-mediated photoreceptor degeneration mechanisms and highlight the importance of PKG as a novel target for the pharmacological intervention in RP. PMID- 19187099 TI - Acetylcholine-induced neuronal differentiation: muscarinic receptor activation regulates EGR-1 and REST expression in neuroblastoma cells. AB - Neurotransmitters are considered part of the signaling system active in nervous system development and we have previously reported that acetylcholine (ACh) is capable of enhancing neuronal differentiation in cultures of sensory neurons and N18TG2 neuroblastoma cells. To study the mechanism of ACh action, in this study, we demonstrate the ability of choline acetyltransferase-transfected N18TG2 clones (e.g. 2/4 clone) to release ACh. Analysis of muscarinic receptors showed the presence of M1-M4 subtypes and the activation of both IP(3) and cAMP signal transduction pathways. Muscarinic receptor activation increases early growth response factor-1 (EGR-1) levels and treatments with agonists, antagonists, and signal transduction enzyme inhibitors suggest a role for M3 subtype in EGR-1 induction. The role of EGR-1 in the enhancement of differentiation was investigated transfecting in N18TG2 cells a construct for EGR-1. EGR-1 clones show increased neurite extension and a decrease in Repressor Element-1 silencing transcription factor (REST) expression: both these features have also been observed for the 2/4 clone. Transfection of this latter with EGR zinc-finger domain, a dominant negative inhibitor of EGR-1 action, increases REST expression, and decreases fiber outgrowth. The data reported suggest that progression of the clone 2/4 in the developmental program is dependent on ACh release and the ensuing activation of muscarinic receptors, which in turn modulate the level of EGR-1 and REST transcription factors. PMID- 19187098 TI - Normobaric hyperoxia attenuates early blood-brain barrier disruption by inhibiting MMP-9-mediated occludin degradation in focal cerebral ischemia. AB - Early blood-brain barrier (BBB) disruption resulting from excessive neurovascular proteolysis by matrix metalloproteinases (MMPs) is closely associated with hemorrhagic transformation events in ischemic stroke. We have shown that normobaric hyperoxia (NBO) treatment reduces MMP-9 increase in the ischemic brain. The aim of this study was to determine whether NBO could attenuate MMP-9 mediated early BBB disruption following ischemic stroke. Rats were exposed to NBO (95% O(2)) or normoxia (30% O(2)) during 90-min middle cerebral artery occlusion, followed by 3-hour reperfusion. NBO-treated rats showed a significant reduction in Evan's blue extravasation in the ischemic hemisphere compared with normoxic rats. Topographically, Evan's blue leakage was mainly seen in the subcortical regions including the striatum, which was accompanied by increased gelatinolytic activity and reduced immunostaining for tight-junction protein, occludin. Increased gelatinolytic activities and occludin protein loss were also observed in isolated ischemic microvessels. Gel gelatin zymography identified that MMP-9 was the main enzymatic source in the cerebral microvessels. Incubation of brain slices or isolated microvessels with purified MMP-9 revealed specific degradation of occludin. Inhibition of MMP-9 by NBO or MMP-inhibitor, BB1101, significantly reduced occludin protein loss in ischemic microvessels. These results suggest that NBO attenuates early BBB disruption, and inhibition of MMP-9-mediated occludin degradation is an important mechanism for this protection. PMID- 19187100 TI - Circulating biomarkers of angiogenesis as indicators of left ventricular systolic dysfunction amongst patients with coronary artery disease. AB - BACKGROUND: Patients with coronary artery disease (CAD) and left ventricular systolic dysfunction (LVSD) are often asymptomatic. Angiogenesis is implicated in the physiology of vascular repair and cardiac remodelling, and is one of many pathophysiological processes implicated in heart failure. We hypothesized that plasma indices associated with angiogenesis [angiogenin, vascular endothelial growth factor (VEGF), and angiopoietin (Ang)-1 and Ang-2] would be abnormal in CAD patients with LVSD, being correlated with EF and wall motion abnormalities (wall motion score) independently of underlying CAD (coronary atheroma score). We also evaluated the specificity of angiogenic 'biomarkers' in their detection of LVSD [ejection fraction (EF) <40%] amongst CAD patients. METHODS: Using a cross sectional approach, we measured angiogenin, VEGF, Ang-1 and Ang-2 by ELISA in 194 CAD patients (aged 34-81 years) undergoing elective coronary angiography. RESULTS: Levels of angiogenin were inversely related with EF (r = -0.17, P = 0.02) and positively with coronary atheroma scores (r = 0.15, P = 0.04, but not independently of EF). Other angiogenic markers were unrelated to objective measures of LVSD but VEGF (P = 0.008) and Ang-2 (P = 0.015) were lower amongst those patients with heart failure. Angiogenin levels were related to wall motion scores (r = 0.16, P = 0.024). CONCLUSION: Heart failure has a modest impact on biomarkers of angiogenesis, in patients with CAD. Further research is warranted into the diagnostic and prognostic utility of biomarkers of angiogenesis, in this common cardiac condition. PMID- 19187101 TI - Feasibility and reliability of body composition measurements in adults with severe intellectual and sensory disabilities. AB - BACKGROUND: Anthropometric measurements are widely used to reliably quantify body composition and to estimate risks of overweight in healthy subjects and in patients. However, information about the reliability of anthropometric measurements in subjects with severe intellectual and sensory disabilities is lacking. OBJECTIVE: The purpose of this study was to determine the feasibility and the test-retest reliability of body composition measures in subjects with severe intellectual and sensory disabilities. METHOD: The study population consisted of 45 subjects with severe intellectual and sensory disabilities. Body mass index, waist circumference, skin folds and tibia length were measured. Reliability was assessed by Wilcoxon signed rank test, limits of agreement (LOA) and intraclass correlation coefficients. The outcomes were compared with values provided by the World Health Organization. RESULTS: There were no significant differences between test and retest (P < 0.05). For the skinfold measurements, however, the LOA was insufficient. Intraclass correlation coefficients for all variables, except skinfold measurements, were 0.90 or above. CONCLUSION: Test retest reliability and feasibility for all measurements are acceptable in subjects with severe intellectual and sensory disabilities. Skinfold measurements, however, could not be reliably performed in these subjects. Measuring tibia length and using the determined formula to calculate body height from tibia length is a reliable alternative for measuring body height. Although measuring the body height of subjects with severe disabilities was feasible, measuring tibia length was more feasible. PMID- 19187102 TI - Behavioural phenotype in Borjeson-Forssman-Lehmann syndrome. AB - BACKGROUND: Borjeson-Forssman-Lehmann syndrome (BFLs) is an X-linked inherited disorder characterised by unusual facial features, abnormal fat distribution and intellectual disability. As many genetically determined disorders are characterised not only by physical features but also by specific behaviour, we studied whether a specific behavioural phenotype exists in BFLs. METHODS: We studied in detail the behaviour of four molecularly proven BFLs patients, and reviewed available literature on BFLs specifically for behavioural characteristics. RESULTS: Behaviour in persons with BFLs is in general friendly, but can be challenging with externalising and thrill-seeking features. Social skills are good. However, variation among patients is wide. Three patients from a single family showed expressed hypersexual behaviour. This was not present in other patients. CONCLUSION: In BFLs a specific behavioural phenotype exists and in behaviour general is challenging besides a friendly habit. Within single families more problematic behaviour may occur. Further behavioural and molecular analysis of a larger group of patients is warranted to determine whether a genotype-behavioural phenotype correlation exists. PMID- 19187103 TI - Impact of coercive tactics on the decision-making of adolescents with intellectual disabilities. AB - BACKGROUND: High rates of victimization have raised concerns about the ability of adolescents with intellectual disabilities (ID) to avoid and escape from harmful situations and to make decisions in their own best interest. The present study was designed to assess the impact of specific coercive tactics on the decision making of adolescents with ID. METHOD: Forty-eight adolescents with ID participated in the study. They were asked to respond to a series of brief vignettes depicting equal numbers of situations involving coercion with a lure, coercion with a threat, and no specific coercive tactic. Performance was assessed in terms of independent, prevention-focused decisions, reporting decisions and responses to fact and inference comprehension questions. RESULTS: Overall, participants suggested independent, prevention-focused decisions only about half the time. They were more likely to suggest independent, prevention-focused decisions in situations with no specific coercive tactic or coercion with a lure than in situations involving a threat. However, reporting decisions were more likely in situations involving coercion with a threat than in the other two conditions and both fact and inference comprehension were best in situations involving coercion with a threat. CONCLUSIONS: Results indicated that adolescents with ID are not well-prepared to handle situations on their own that involve coercion, especially coercion with a threat. Because comprehension did not appear to be a key source of the decision-making difficulty in this study, further research is needed to examine all aspects of the decision-making process as a basis for the design of effective interventions. PMID- 19187104 TI - An infiltrative variant of non-neural granular cell tumor: a case report. AB - Dermal non-neural granular cell tumors are rare tumors of indeterminate lineage that typically present as well-circumscribed tumors with nuclear pleomorphism and mitotic activity. We describe a dermal non-neural granular cell tumor with a distinctive growth pattern with granular cells interspersed between collagen bundles. This asymptomatic papule arose on the scapula of a 46-year-old woman and consisted of a mixture of epithelioid and spindled granular cells. The immunohistochemical characteristics were similar to those of previously reported dermal non-neural granular cell tumors. Despite mild nuclear pleomorphism and dispersion of lesional cells among collagen bundles, mitoses were not present and Ki-67 staining indicated a low proliferative rate. In addition to being S-100 protein negative and NKI/C3 positive, our case was positive for PGP9.5 and weakly positive for neuron-specific enolase, a staining pattern similar to what has been observed for cellular neurothekeomas. Our case could represent a dermal non neural granular cell tumor with unique architecture, a granular cellular neurothekeoma or a granular cell dermatofibroma. As both dermal non-neural granular cell tumor and cellular neurothekeoma are of indeterminate lineage, our case with features characteristic of both entities may suggest a common precursor or lineage for dermal non-neural granular cell tumor and cellular neurothekeoma. PMID- 19187105 TI - Localized eosinophilic (Langerhans' cell) granuloma of the lower lip. A lesion that may cause diagnostic error. AB - Eosinophilic (Langerhans' cell) granuloma is most common in children and is unusual in the elderly. A cutaneous lesion as the sole manifestation of adult Langerhans' cell histiocytosis is rare. We report an 81-year-old man who presented with a noduloulcerative lesion on the lower lip. A punch biopsy suggested a lymphoepithelioma-like carcinoma. In the absence of immunohistochemistry, the large pale cells with atypical lobular nuclei and patent nucleoli, with an overlying ulceration, were interpreted as carcinoma cells. Definitive diagnosis was established after complete resection of the lesion aided with the use of immunohistochemistry. The proliferating cells were S100, CD1a and CD207 (langerin) positive and cytokeratin, epithelial membrane antigen, CD15, CD30, melan A and carcinoembryonic antigen negative. To our knowledge, eosinophilic (Langerhans' cell) granuloma of the lip has not been described. The case constituted a diagnostic challenge because of its unusual clinical presentation and its histopathological similarity to certain other benign and malignant entities. Because there was no other organ involved, the patient was treated with complete surgical excision without recurrence or other organ involvement in the 4-month follow-up period. Labial eosinophilic (Langerhans' cell) granuloma should be considered in the histological differential diagnosis in cases with atypical polymorphous infiltrates including large pale cells. PMID- 19187106 TI - Janeway lesions with inconspicuous histological features. AB - Septic-embolic diseases are life threatening; survival critically depends on rapid diagnosis. Clinically, cutaneous manifestations like Janeway lesions are infrequently occurring but highly characteristic. In contrast, histological features are not clearly defined. We report the case of a 59-year-old male patient in deteriorating general condition and fever of unknown origin with lesions suggestive for Janeway lesions. Histology showed dilated blood vessels and extravasated erythrocytes throughout the dermis. Only serial sections revealed single, deep dermal vessels occluded by fibrin thrombi. Computed tomography scan identified multiple pulmonary round masses. Blood cultures showed growth of Staphylococcus aureus and Enterococcus faecalis. The knowledge of the described subtle histological features and a clinical-pathological correlation is essential for the diagnosis of certain cases of septic-embolic conditions. PMID- 19187107 TI - Caveats in BerEP4 staining to differentiate basal and squamous cell carcinoma. AB - BACKGROUND: Superficial skin biopsies of basal cell carcinoma (BCC) represent some of the most common dermatopathology specimens. Superficial shave biopsies containing partial samples of lesions with squamatization present difficulties in distinguishing BCC from squamous cell carcinoma (SCC). BerEP4 has been employed as a dependable marker in differentiating between BCCs and SCCs. METHODS: We collected 12 cases of superficial biopsies of BCC with centrally located cords and strands suggesting squamous differentiation at the Yale Dermatopathology Laboratory over a 3-month period and stained them with BerEP4. RESULTS: We found that all cases (12 out of 12) showed membranous and cytoplasmic staining with BerEP4 in the obvious areas of basaloid differentiation at the periphery of the tumors, while cords and strands of enlarged cells with squamoid features in the center and surface of the biopsy failed to label with BerEP4. CONCLUSIONS: BerEP4 labeling is not reliable in superficial biopsies of BCC with squamoid features. It is important to be aware of this caveat in interpreting BerEP4 staining for BCC and SCC. PMID- 19187108 TI - Angiokeratoma-like changes in extragenital and genital lichen sclerosus. AB - Hemorrhagic blisters have rarely been described developing in the background of either genital or extragenital lichen sclerosus and have invariably been designated clinically as telangiectatic, hemorrhagic or bullous lichen sclerosus. We describe three patients with extragenital and genital lichen sclerosus, who presented clinically with hemorrhagic plaques and/or papules. In addition to the classical histology of lichen sclerosus, dilated, congested and focally thrombosed vascular channels lined by flat endothelium were seen within the sclerotic dermal collagen. They were in close proximity to and even in contact with the overlying epidermis and thus mimicked an angiokeratoma. Angiokeratoma like changes in lichen sclerosus represent secondary features because of damage to the dermis by lichen sclerosus and are characterized histologically by ectatic thin-walled vascular spaces in the papillary dermis intimately associated with the epidermis. Increased venous pressure, local trauma, degenerative changes in the elastic tissue of the vessel wall and/or surrounding supportive tissue, as well as abnormalities in the extracellular protein network, appear to be implicated in their pathogenesis. PMID- 19187110 TI - IgG4-positive plasma cells in cutaneous Rosai-Dorfman disease: an additional immunohistochemical feature and possible relationship to IgG4-related sclerosing disease. AB - BACKGROUND: Cutaneous Rosai-Dorfman disease (CRDD) shares the histopathological features of abundant plasma cells and stromal fibrosis with IgG4-related sclerosing disease. The possible role of IgG4+ plasma cells in CRDD was investigated. METHODS: Twelve cases of CRDD were reviewed, and their lesions were immunostained with anti-IgG4 and anti-IgG antibodies. The number of IgG4+ and IgG+ plasma cells and their ratios were estimated. Serum IgG4 and IgG concentrations were measured in two recent cases. RESULTS: Many IgG4+ and IgG+ plasma cells were found in all 12 cases. IgG4+ plasma cells ranged from 21 to 204 per high-power field (HPF) (mean 117/HPF), and IgG+ plasma cells ranged from 114 to 759/HPF (mean 349/HPF). All cases had more than 30 IgG4+ cells/HPF, except one case. The IgG4/IgG ratio ranged from 16% to 51% (mean 34%). Serum IgG4 concentration and serum IgG4/IgG ratio were increased in one recent case. Various degrees of stromal fibrosis were present in all cases. CONCLUSIONS: The presence of many IgG4+ plasma cells and stromal fibrosis suggests that CRDD may be related to IgG4-related sclerosing disease. Many IgG4+ plasma cells is another feature of CRDD, and serum IgG4 may be elevated. PMID- 19187111 TI - Melanocytic nevi with features of Spitz nevi and Clark's/dysplastic nevi ("Spark's" nevi). AB - BACKGROUND: Nevi with cytologic characteristics of Spitz nevus and architectural features of Clark's/dysplastic nevus are not well recognized in the literature. METHODS: Twenty-seven nevi with characteristics of Spitz nevus and Clark's/dysplastic nevus are reviewed. RESULTS: The patients' mean age was 33 years, and 17/27(63%) patients were female. Lesions were most frequent on the trunk and lower extremities. Histopathologically, these nevi were composed of large, monomorphous spindled and/or epithelioid melanocytes. Spindle cells were often oriented parallel to the epidermis, with fused rete and lamellar fibroplasias. Lateral extension of the junctional component was a feature of compound lesions. An average of 10 years of clinical follow up in 12 patients revealed no recurrence or metastasis. CONCLUSIONS: Recognition of this type of nevus is important to avoid confusion with malignant melanoma. PMID- 19187112 TI - Analysis of cell cycle phases and proliferative capacity in mice bearing melanoma maintained on different dietary proteins. AB - BACKGROUND: Diet seems to represent, directly or indirectly, 35% of all cancer reports. In this study, the influence of dietary protein on the growth of melanoma B16F10 was evaluated through analyses of cell cycle phases and proliferative capacity. METHODS: Flow cytometry and argyrophilic nucleolar organizer regions (AgNORs) technique were applied in mice bearing B16F10 melanoma cells fed on different dietary proteins. All data were submitted to statistical analyses. RESULTS: The G0/G1 phase increased for the animal groups fed bovine collagen hydrolysate (BCH) or BCH-P1 + whey protein isolate (WPI), compared with mice receiving only WPI, for all dietary groups treated and nontreated with paclitaxel. Mice that received BCH + WPI treated with paclitaxel showed the highest percentage of apoptosis compared with WPI group. AgNORs, total nucleolar organizer regions (NORs)/cells and dot number/cell for all dietary protein groups nontreated with paclitaxel were higher than for the WPI. The only two dietary protein groups treated with paclitaxel that presented higher total NORs and dot number/cell than the WPI group were BCH + WPI and BCH-P1 + WPI. CONCLUSIONS: A significantly lower proliferative capacity and larger number of cells in the G0/G1 phase were observed for the dietary protein groups combining the two collagen hydrolysates, BCH or BCH-P1 with WPI, treated with paclitaxel. PMID- 19187113 TI - Infiltrating intramuscular spindle cell lipoma of the face. AB - Spindle cell lipoma is a benign lipomatous tumor, which usually arises on the back of the neck, shoulder or upper back of males in the third to seventh decade of life. We report herein an unusual infiltrating intramuscular spindle cell lipoma arising in the nose of a 53-year-old man. The patient presented with a 0.5 cm 'cyst' of the nose, just above the right alar crease, which was removed. Four years later, the lesion recurred and was re-excised. Histologically, a proliferation of mature adipocytes, ropey collagen fibers and spindle cells within a myxoid stroma was present in the subcutaneous tissue and infiltrated between skeletal muscle fibers. Nine cases of intramuscular spindle cell lipoma with histological examination have previously been reported and have involved the oral cavity and muscles of the extremities. To our knowledge, this is the first report of an infiltrating intramuscular spindle cell lipoma arising on the face. PMID- 19187114 TI - Multiple cutaneous sclerosing perineuriomas: an extensive presentation with involvement of the bilateral upper extremities. AB - A perineurioma is a benign peripheral nerve sheath neoplasm thought to be derived from cells of the perineurium, the cellular sheathing of peripheral nerve fascicles. The histological, immunohistochemical (positive for epithelial membrane antigen, collagen type IV, laminin and vimentin but not S-100 protein) and ultrastructural features of a perineurioma form the basis for diagnosis. Two types of perineuriomas are recognized, intraneural and soft tissue (extraneural) forms. Sclerosing perineurioma is considered a variant of the soft tissue perineurioma. In all but one of the reported cases, this subtype presented as a solitary clinical lesion with a predilection for the fingers and palms of young adults. In 2002, Huang and Sung described a 16-year-old boy with one sclerosing perineurioma on each hand. The unusual case presented herein is a 21-year-old woman manifesting more than 30 cutaneous sclerosing perineuriomas on the hands and arms. This case is the most extensive presentation of sclerosing perineuriomas documented to date and extends the clinical presentation spectrum of these lesions. PMID- 19187115 TI - Solitary superficial angiomyxoma: an infrequent but distinct soft tissue tumor. AB - Superficial angiomyxoma (SA) is a distinct soft tissue tumor characterized by a circumscribed collection of spindled and stellate fibroblasts that are admixed with thin-walled blood vessels and embedded in a mucinous stroma. Because of its relative infrequent occurrence, the purpose of this article was to present a classical example of an isolated superficial angiomyxoma and discuss the differential diagnosis. PMID- 19187116 TI - Expressions of p53, cyclinD1 and histopathological features in basal cell carcinomas. AB - BACKGROUND: We planned this study to analyze probable associations between p53, cyclinD1, Ki67 and histopathological features in basal cell carcinomas (BCC). METHODS: Histological differentiation types, histological growth patterns and tissue responses were analyzed in 50 cases of BCC. In immunohistochemical analysis, p53, cyclinD1 and Ki67 antibodies were investigated. P53 expression was evaluated based on a cut-off value of 25% positivity. CyclinD1 expression was graded from 0 to 3+ according to the percentage of positive nuclear staining. The percentage of positively staining cells for Ki67 was recorded. RESULTS: The following significant correlations were detected. Solid infiltrative type differentiation was related to the infiltrative histological growth pattern. The rates of p53 positivity and severe fibrosis in the groups of mixed and infiltrative growth patterns were higher than others. Besides, p53-positive cases showed more severe fibrosis and had a higher mean value for Ki67 index. Epidermal p53 and cyclinD1 clones in normal epidermal areas adjacent to tumors were noticed in 42% and 52% of the cases, respectively. CONCLUSIONS: P53 expression seems to be related to Ki67 index and some histopathological features of BCC, such as infiltrative histological growth pattern and probably fibrosis. PMID- 19187117 TI - Skin biopsy for inflammatory and common neoplastic skin diseases: optimum time, best location and preferred techniques. A critical review. AB - The diagnosis of skin diseases, particularly inflammatory dermatoses, is based primarily on clinical information. Pathologic examination of the biopsied specimen often serves as a complementary or confirmative part of the diagnosis. However, the clinical diagnosis of skin diseases may be challenging, as the clinical information and appearance of skin lesions invariably overlap. Evidence for a correct diagnosis may be lacking without histopathologic examination of skin biopsies. It is well known that the histologic diagnosis of inflammatory and other skin diseases requires clinicopathologic correlation, and there is evolution of skin lesions into different stages as the diseases progress. Other factors important for accurate dermatopathologic diagnosis are optimum time, best location and preferred techniques of skin biopsy. In searching for available information concerning when, where and how to take skin biopsies, it is noted that there are only limited practical guidelines currently available. We present this review article in hopes that our collective dermatopathologic and dermatologic experience can provide a quick reference for accurate diagnosis and proper management of skin diseases. PMID- 19187127 TI - Presence of thyrotropin receptor in hepatocytes: not a case of illegitimate transcription. AB - The function of thyrotropin (TSH) in the thyroid gland is mediated by thyrotropin receptor (TSHR). In addition to the thyroid, TSHR expression has been described in some non-thyroidal tissues, although it is uncertain whether TSHR is present in hepatocytes. One study has reported hepatic expression of TSHR mRNA, but this was considered to be because of illegitimate transcription, and there has not been a study investigating its protein expression and function in hepatocytes. Here, we examined the expression of TSHR in human and rat liver tissues, as well as human normal hepatocyte cell line L-02. Our results demonstrated that hepatic TSHR mRNA could be detected and had the same sequence as that of thyroid-derived mRNA. TSHR protein was also expressed and mainly located in the hepatocyte cell membrane. Moreover, bovine TSH and immunoglobulin from sera of patients with Graves' disease stimulated cAMP production in these cells. Taken together, these data show that TSHR is present and functional in hepatocytes, and this expression is not a case of illegitimate transcription. Given the pivotal role of the liver in body metabolism and many human diseases, our findings provide important implications for a potentially novel physiopathological role of TSH via acting on the TSHR in hepatocytes besides its classical role in regulating the thyroid function. PMID- 19187128 TI - Functional expression of human HMG-CoA reductase in Saccharomyces cerevisiae: a system to analyse normal and mutated versions of the enzyme in the context of statin treatment. AB - AIMS: Statins - inhibitors of the 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase - are known to reduce blood cholesterol levels. In this paper, we present a Saccharomyces cerevisiae expression system, which enables quick evaluation of the sensitivity of the wild-type and/or mutant forms of human HMG CoA reductase towards statins or other drugs. METHODS AND RESULTS: We analysed the sequence of the HMG-CoA reductase gene in DNA extracted from blood samples of 16 patients with cardiovascular disorders. We applied the yeast system to examine the sensitivity of the wild-type and mutated versions of the hHMG-CoA reductase to different types of statins. CONCLUSION: The yeast and mammalian HMG-CoA reductases demonstrate structural and functional conservation, and expression of human HMG-CoA reductase in yeast complements the lethal phenotype of strains lacking the HMG1 and HMG2 genes. SIGNIFICANCE AND IMPACT OF THE STUDY: These data indicate that a yeast expression system can serve to study the influence of selected mutations in human HMG-CoA reductase on the sensitivity of the enzyme to commonly prescribed statins. Our results suggest that this model system is suitable for the development and selection of lipid-lowering drugs as well as for the examination of DNA sequence variations in the context of statin therapy. PMID- 19187129 TI - Sphagnan--a pectin-like polymer isolated from Sphagnum moss can inhibit the growth of some typical food spoilage and food poisoning bacteria by lowering the pH. AB - AIMS: Investigate if the antibacterial effect of sphagnan, a pectin-like carbohydrate polymer extracted from Sphagnum moss, can be accounted for by its ability to lower the pH. METHODS AND RESULTS: Antibacterial activity of sphagnan was assessed and compared to that of three other acids. Sphagnan in its acid form was able to inhibit growth of various food poisoning and spoilage bacteria on low buffering solid growth medium, whereas sphagnan in its sodium form at neutral pH had no antibacterial activity. At similar acidic pH, sphagnan had comparable antibacterial activity to that of hydrochloric acid and a control rhamnogalacturonan pectin in its acid form. CONCLUSIONS: Sphagnan in its acid form is a weak macromolecular acid that can inhibit bacterial growth by lowering the pH of environments with a low buffering capacity. SIGNIFICANCE AND IMPACT OF THE STUDY: It has previously been suggested that sphagnan is an antimicrobial polysaccharide in the leaves of Sphagnum moss with a broad range of potential practical applications. Our results now show that sphagnan in its acid form can indeed inhibit bacterial growth, but only of acid-sensitive species. These findings represent increased knowledge towards our understanding on how sphagnan or Sphagnum moss might be used in practical applications. PMID- 19187130 TI - The efficacy of biocides and other chemical additives in cooling water systems in the control of amoebae. AB - AIMS: In vitro experiments were undertaken to evaluate biocide formulations commonly used in cooling water systems against protozoa previously isolated from cooling towers. The investigations evaluated the efficacy of these formulations against amoebic cysts and trophozoites. METHODS AND RESULTS: Laboratory challenges against protozoa isolated from cooling towers using chlorine, bromine and isothiazolinone biocides showed that all were effective after 4 h. The presence of molybdate and organic phosphates resulted in longer kill times for bromine and isothiazolinones. All treatments resulted in no detectable viable protozoa after 4 h of exposure. CONCLUSIONS: The chemical disinfection of planktonic protozoa in cooling water systems is strongly influenced by the residence time of the formulation and less so by its active constituent. Bromine and isothiazolinone formulations may require higher dosage of concentrations than currently practiced if used in conjunction with molybdate- and phosphate-based scale/corrosion inhibitors. SIGNIFICANCE AND IMPACT OF THE STUDY: Cooling water systems are complex microbial ecosystems in which predator-prey relationships play a key role in the dissemination of Legionella. This study demonstrated that at recommended dosing concentrations, biocides had species-specific effects on environmental isolates of amoebae that may act as reservoirs for Legionella multiplication in cooling water systems. PMID- 19187131 TI - Indigestible carbohydrates alter the intestinal microbiota but do not influence the performance of broiler chickens. AB - AIMS: Prebiotics are a potential alternative to in-feed antimicrobials to improve performance of chickens. We investigated the effects of mannanoligosaccharide (MOS) and fructooligosaccharide (FOS) on growth, performance and the intestinal microbiota. METHODS AND RESULTS: Cobb 500 birds were fed either: Control, starter diet without antimicrobials; ZnB, Control + 50 ppm zinc bacitracin; MOS, Control + 5 g kg(-1) MOS; or FOS, Control + 5 g kg(-1) FOS. An energy metabolism study was conducted and intestinal microbial communities assessed by T-RFLP and Lac PCR DGGE. Diet did not influence performance. Ileal microbial communities were significantly different in ZnB-fed birds compared to all diets, and FOS-fed chickens compared to Control. MOS-fed chickens had a different caecal profile to ZnB and FOS-fed birds. Consensus Lac PCR-DGGE profiles indicated Lactobacillus communities clustered according to diet with Lactobacillus johnsonii characteristic of ZnB diet. Control and MOS-fed chickens displayed significantly different jejunal Lactobacillus profiles to each other whilst ileal profiles were different between MOS and FOS-fed birds. CONCLUSION: Prebiotics influenced the intestinal microbiota, but did not affect performance. SIGNIFICANCE AND IMPACT OF THE STUDY: In light of pressure for in-feed antimicrobial withdrawal, the impact of alternative compounds on the intestinal microbiota and bird performance is critical to the poultry industry. PMID- 19187132 TI - Distribution and diversity of the enterococcal surface protein (esp) gene in animal hosts and the Pacific coast environment. AB - AIMS: This study sought to evaluate the distribution of the enterococcal surface protein (esp) gene in Enterococcus faecium in the Pacific coast environment as well as the distribution and diversity of the gene in Northern California animal hosts. METHODS AND RESULTS: Over 150 environmental samples from the Pacific coast environment (sand, surf zone, fresh/estuarine, groundwater, and storm drain) were screened for the esp gene marker in E. faecium, and the marker was found in 37% of the environmental samples. We examined the host specificity of the gene by screening various avian and mammalian faecal samples, and found the esp gene to be widespread in nonhuman animal faeces. DNA sequence analysis performed on esp polymerase chain reaction amplicons revealed that esp gene sequences were not divergent between hosts. CONCLUSIONS: Our data confirm recent findings that the E. faecium variant of the esp gene is not human-specific. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results suggest that the use of the esp gene for microbial source tracking applications may not be appropriate at all recreational beaches. PMID- 19187133 TI - The potential use of chilling to control the growth of Enterobacteriaceae on porcine carcasses and the incidence of E. coli O157:H7 in pigs. AB - AIMS: To (i) monitor the presence of Enterobacteriaceae as indicators of faecal contamination on pig carcasses, (ii) examine the potential use of chilling as a critical control point (CCP) and establish its influence on pig carcass categorization by Decision 471/EC and (iii) determine the incidence of E. coli O157:H7 in pigs. METHODS AND RESULTS: Porcine faecal samples and carcass swabs were collected before and after chilling at four Irish pig abattoirs and examined for Enterobacteriaceae and E. coli O157:H7. Chilling generally reduced Enterobacteriaceae counts on carcasses, but increases were also observed, particularly in one abattoir. E. coli O157:H7 was absent from carcasses before chilling, present on 0.21% after chilling and was recovered from 0.63% of faecal samples. All of the isolates were found to contain virulence genes associated with clinical illness in humans. CONCLUSIONS: The data show that overall chilling had the capacity to reduce the numbers of carcasses positive for the presence of Enterobacteriaceae. SIGNIFICANCE AND IMPACT OF STUDY: The influence of chilling on the categorization of pig carcasses suggests that it has the potential to improve the numbers of acceptable carcasses and the process could be used as a CCP within a HACCP plan. PMID- 19187134 TI - Diversity and spatial distribution of amoA-encoding archaea in the deep-sea sediments of the tropical West Pacific Continental Margin. AB - AIMS: The ecological characteristics of the deep-sea amoA-encoding archaea (AEA) are largely unsolved. Our aim was to study the diversity, structure and distribution of the AEA community in the sediments of the tropical West Pacific Continental Margin, to develop a general view of the AEA biogeography in the deep sea extreme environment. METHODS AND RESULTS: Archaeal amoA clone libraries were constructed. Diverse and novel amoA sequences were identified, with the Bohol Sea, Bashi Strait and Sibuyan Sea harbouring the highest and the Bicol Shelf the lowest AEA diversity. Phylogenetic and statistical analyses illustrate a heterogeneous distribution of the AEA community, probably caused by the differential distribution of the terrestrial or estuarine AEA in the various sampling sites. CONCLUSIONS: The deep-sea sedimentary environments potentially harbour diverse and novel AEA in the tropical West Pacific Continental Margin. The stations in the Philippine inland seas (including station 3043) may represent AEA assemblages with various terrestrial influences and the stations connected directly to the open Philippine Sea may represent marine environment-dominant AEA assemblages. SIGNIFICANCE AND IMPACT OF STUDY: Our study indicates the potential importance of geological and climatic events in the transport of terrestrial micro-organisms to the deep-sea sedimentary environments, almost totally neglected previously. PMID- 19187135 TI - Virulence of luminescent and non-luminescent isogenic vibrios towards gnotobiotic Artemia franciscana larvae and specific pathogen-free Litopenaeus vannamei shrimp. AB - AIMS: This study was conducted to test the virulence of luminescent (L) and non luminescent (NL) isogenic strains of Vibrio campbellii LMG21363, Vibrio harveyi BB120 (wild type) and quorum-sensing mutant strains derived from the wild type such as Vibrio harveyi BB152, BB170, MM30 and BB886. METHODS AND RESULTS: The NL strains could be obtained by culturing rifampicin-resistant luminescent strains in the dark under static condition. The virulence of the L and NL strains was tested in gnotobiotic Artemia franciscana larvae challenged with 10(4) CFU ml(-1) of bacteria. All luminescent isogenic tested strains showed higher virulence compared to the NL strains. The virulence of L and NL V. campbellii and V. harveyi BB120 was also tested in specific pathogen-free juvenile shrimp upon intramuscular injection with 10(6) CFU of bacteria. In contrast with Artemia, there was no significant difference in mortality between the groups challenged with L and NL strains (P > 0.05). The non-luminescent strains were not able to revert back to the luminescent state and quorum sensing did not influence this phenotypic shift. CONCLUSIONS: Luminescent Vibrio strains can switch to a non luminescent state by culturing them in static conditions. The NL strains become less virulent as verified in Artemia. SIGNIFICANCE AND IMPACT OF THE STUDY: The luminescent state of Vibrio cells in a culture needs to be verified in order to assure maintenance of virulence. PMID- 19187137 TI - Comparative analysis of different TaqMan real-time RT-PCR assays for the detection of swine Hepatitis E virus and integration of Feline calicivirus as internal control. AB - AIMS: The aim of this study was to compare the performance of four TaqMan RT-PCR assays with a commonly used nested RT-PCR and to include the Feline calicivirus (FCV) as an internal control. METHODS AND RESULTS: RNA extracted from 87 swine faecal samples and 103 swine blood samples was subjected to different detection systems. Faecal samples naturally contaminated with Hepatitis E virus (HEV) and negative samples were artificially inoculated with 3.2 x 10(3) PFU of FCV. Detection results obtained on faecal and plasma samples were 35.6% and 4.9% with the nested RT-PCR assay, 8.0% and 0%, 0% and 0%, 13.8% and 0% and 36.8% and 3.9% with TaqMan systems A, B, C and D respectively. The Ct means obtained with the multiplex TaqMan assay were 30.11 and 30.43 for the detection of FCV with HEV contaminated samples and negative samples. CONCLUSIONS: The TaqMan system D was more suitable for the detection of swine HEV strains than the three others and FCV was integrated successfully as an internal control. SIGNIFICANCE AND IMPACT OF THE STUDY: FCV was demonstrated as an efficient control to monitor the RNA extraction process and HEV amplification procedure in a multiplex HEV/FCV TaqMan assay. This control would be helpful in limiting false negative results. PMID- 19187138 TI - The mechanism of action of a citrus oil blend against Enterococcus faecium and Enterococcus faecalis. AB - AIMS: The aim was to explore the mechanisms by which a blend of orange (Citrus sinensis) : bergamot (Citrus bergamia) (1 : 1 v/v) EO (essential oil) (2% v/v) and its vapour (15 mg l(-1) air) brings about its antimicrobial effect against Enterococcus faecium and Enterococcus faecalis. METHODS AND RESULTS: Cells were exposed to the blend in oil or vapour form in a sealed unit. Membrane permeability was measured using an NPN assay and intra and extracellular ATP concentrations were assessed using luminescence. Assays using 3,3 dipropylthiacarbocyanine and carboxyfluorescein diacetate succinimidyl ester measured membrane potential and intracellular pH changes. TEM images of treated cells indicate morphological differences and show the possible uptake of the EO into the cell. After cells were exposed to EO or vapour, cell permeability increased by x2 and x40 respectively. A decrease of 1.5 in intracellular pH, 20 a.u. in membrane potential and 18 pmol mg(-1) protein of intracellular ATP occurred. CONCLUSIONS: The EO blend affects the cell membrane and cell homeostasis resulting in inhibition of growth or cell death. SIGNIFICANCE AND IMPACT OF THE STUDY: Understanding the mechanisms by which EOs bring about their antibacterial effect could lead to an alternative to chemical-based bactericides for use against Enterococcus sp. PMID- 19187136 TI - Bacterial diversity associated with the Brazilian endemic reef coral Mussismilia braziliensis. AB - AIMS: We performed the first characterization of the microbiota associated with the reef coral Mussismilia braziliensis by means of a culture-independent approach. METHODS AND RESULTS: The main groups were Proteobacteria, Cyanobacteria and unclassified bacteria according to the 16S rDNA libraries. Most of the sequences of the mucus of healthy and diseased M. braziliensis did not find close matches in GenBank (i.e. >97% 16S rDNA similarity). Most of the sequences of seawater and mucus of healthy coral fell into tight clusters (17 and 15 clusters respectively). In contrast, most of the sequences of mucus of diseased coral did not form clusters. The rarefaction curves indicate saturation in the recovery of higher taxa (approximately 40 phyla). However, the number of species in the coral mucus (n = 130-170) and seawater (n = 170) did not reach a plateau. CONCLUSIONS: The coral microbiota encompasses several potentially novel species and higher taxa. The microbiota of M. braziliensis appears to be species-specific. Diseased coral may have provided a suitable place for colonization by opportunistic bacteria, resulting in a greater bacterial diversity. SIGNIFICANCE AND IMPACT OF THE STUDY: The first study on the diversity of the microbiota of the endemic and endangered of extinction coral M. braziliensis. PMID- 19187139 TI - Sphingobacterium sp. strain PM2-P1-29 harbours a functional tet(X) gene encoding for the degradation of tetracycline. AB - AIMS: The tet(X) gene has previously been found in obligate anaerobic Bacteroides spp., which is curious because tet(X) encodes for a NADP-dependent monooxygenase that requires oxygen to degrade tetracycline. In this study, we characterized a tetracycline resistant, aerobic, Gram-negative Sphingobacterium sp. strain PM2-P1 29 that harbours a tet(X) gene. METHODS AND RESULTS: Sphingobacterium sp. PM2-P1 29 demonstrated the ability to transform tetracycline compared with killed controls. The presence of the tet(X) gene was verified by PCR and nucleotide sequence analysis. Additional nucleotide sequence analysis of regions flanking the tet(X) gene revealed a mobilizable transposon-like element (Tn6031) that shared organizational features and genes with the previously described Bacteroides conjugative transposon CTnDOT. A circular transposition intermediate of the tet(X) region, characteristic of mobilizable transposons, was detected. However, we could not demonstrate the conjugal transfer of the tet(X) gene using three different recipient strains and numerous experimental conditions. CONCLUSIONS: This study suggests that Sphingobacterium sp. PM2-P1-29 or a related bacterium may be an ancestral source of the tet(X) gene. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates the importance of environmental bacteria and lateral gene transfer in the dissemination and proliferation of antibiotic resistance among bacteria. PMID- 19187140 TI - Biofilm problems in dental unit water systems and its practical control. AB - Dental chair units (DCUs) contain integrated systems that provide the instruments and services for a wide range of dental procedures. DCUs use water to cool and irrigate DCU-supplied instruments and tooth surfaces during dental treatment. Water is supplied to these instruments by a network of interconnected narrow-bore (2-3 mm) plastic tubes called dental unit waterlines (DUWLs). Many studies over the last 40 years demonstrated that DUWL output water is often contaminated with high densities of micro-organisms, predominantly Gram-negative aerobic heterotropic environmental bacteria, including Legionella and Pseudomonas species. Untreated DUWLs host biofilms that permit micro-organisms to multiply and disperse through the water network and which are aerosolized by DCU instrument use, thus exposing patients and staff to these micro-organisms, to fragments of biofilm and bacterial endotoxins. This review concentrates on how practical developments and innovations in specific areas can contribute to effective DUWL biofilm control. These include the use of effective DUWL treatment agents, improvements to DCU supply water quality, DCU design changes, development of automated DUWL treatment procedures that are effective at controlling biofilm in the long-term and require minimal human intervention, are safe for patients and staff, and which do not cause deterioration of DCU components following prolonged use. PMID- 19187141 TI - Influence of phosphorus on biofilm formation in model drinking water distribution systems. AB - AIMS: This study investigated the effects of phosphorus on biofilm formation via annular reactor systems in terms of biofilm cell growth, exopolysaccharide (EPS) production, biofilm structure and cell metabolic potential. METHODS AND RESULTS: Drinking water biofilms were developed in annular reactors with supplement of carbon and different levels of phosphorus. The biofilm formation was monitored over a period of 30 days. Biofilm related parameters were examined by various methods, which included heterotrophic plate count, total carbohydrate content, confocal laser scanning microscopy and GN2 microplate assay. Our results showed that phosphorus addition can promote the biofilm cell growth (cell count increased about 1 log with addition of 30 and 300 microg l(-1) of phosphorus). However, the addition of 30 and 300 microg l(-1) of phosphorus caused 81% and 77% decrease in EPS production, respectively. The results of biofilm structure analysis showed that the addition of 30 and 300 microg l(-1) of phosphorus can induce thicker and less homogeneous biofilms with more biomass. Furthermore, the addition of 30 and 300 microg l(-1) of phosphorus dramatically increased the biofilm cell metabolic potential. The addition of 3 microg l(-1) of phosphorus was found to have minor effects on the parameters examined. CONCLUSIONS: The results indicate phosphorus addition to drinking water distribution system (DWDS) has a complicated effect on the biofilm formation. SIGNIFICANCE AND IMPACT OF THE STUDY: As the addition of phosphorus at certain levels can affect the biofilm growth in DWDS, care should be taken when phosphate-based corrosion inhibitors are used in the DWDS. PMID- 19187142 TI - Optimization of a reconstituted skim milk based medium for enhanced CLA production by bifidobacteria. AB - AIMS: To determine the effect of a range of supplements on the bioconversion of linoleic acid to conjugated linoleic acid (CLA) by Bifidobacterium breve NCIMB 702258 in reconstituted skim milk (RSM). RESULTS: Seven supplements (yeast extract, casein hydrolysate, tryptone, l-cysteine hydrochloride, sodium acetate, sodium butyrate and sodium propionate) were identified as increasing the bioconversion of linoleic acid to c9, t11 CLA. Using these supplements, the percentage bioconversion of linoleic acid (0.35 mg ml(-l)) to the c9, t11 CLA isomer was elevated from 15.5 +/- 1.1% in 20% RSM (w/v) to 48.1 +/- 2.2% in the supplemented RSM. Through additional supplementation of 20 mg m1(-1) inulin and optimization of inoculum and linoleic acid concentration, the percentage bioconversion to c9, t11 CLA was increased to 55.0 + 3.2%. CONCLUSIONS: Through supplementation, the concentration of CLA produced by bifidobacteria in RSM can be increased to levels comparable to those observed in the synthetic medium cys MRS. SIGNIFICANCE AND IMPACT OF THE STUDY: The impact of 22 supplements on the production of the c9, t11 CLA isomer by the strain B. breve NCIMB 702258 in milk has been determined. The results provide an understanding of the factors, which influence CLA production by bifidobacteria in RSM. PMID- 19187143 TI - Biological sulfide oxidation using autotrophic Thiobacillus sp.: evaluation of different immobilization methods and bioreactors. AB - AIMS: Evaluation of various immobilization methods and bioreactors for sulfide oxidation using Thiobacillus sp. was studied. METHODS AND RESULTS: Ca-alginate, K carrageenan and agar gel matrices (entrapment) and polyurethane foam and granular activated carbon (adsorption) efficacy was tested for the sulfide oxidation and biomass leakage using immobilized Thiobacillus sp. Maximum sulfide oxidation of 96% was achieved with alginate matrix followed by K-carrageenan (88%). Different parameters viz. alginate concentration (1%, 2%, 3%, 4% and 5%), CaCl(2) concentration (1%, 2%, 3%, 4% and 5%), bead diameter (1, 2, 3, 4 and 5 mm), and curing time (1, 3, 6, 12 and 18 h) were studied for optimal immobilization conditions. Repeated batch experiments were carried out to test reusability of Ca alginate immobilized beads for sulfide oxidation in stirred tank reactor and fluidized bed reactor (FBR) at different sulfide concentrations. CONCLUSIONS: The results proved to be promising for sulfide oxidation using Ca-alginate gel matrix immobilized Thiobacillus sp. for better sulfide oxidation with less biomass leakage. SIGNIFICANCE AND IMPACT OF THE STUDY: Biological sulfide oxidation is gaining more importance because of its simple operation. Present investigations will help in successful design and operation of pilot and industrial level FBR for sulfide oxidation. PMID- 19187144 TI - Molecular source tracking of predominant lactic acid bacteria in traditional Belgian sourdoughs and their production environments. AB - AIMS: To investigate the circulation of predominant sourdough lactic acid bacteria (LAB) species in the production environment of two Belgian artisan sourdough bakeries. METHODS AND RESULTS: Isolates were collected from sourdoughs, flour, hands of the baker and air in the bakery setting and taxonomically characterized using repetitive element sequence-based PCR fingerprinting, pheS and/or 16S rRNA gene sequencing and amplified fragment length polymorphism (AFLP) analysis. In parallel, PCR-DGGE (denaturing gradient gel electrophoresis) analysis of V3-16S rDNA amplicons was applied to visualize the predominant bacterial population in the sourdoughs and the corresponding bakery environment (flour, hands of the baker, air and bakery equipment). Both approaches revealed that sourdoughs produced at D01 and D10 were mainly dominated by Lactobacillus spicheri and L. plantarum and by L. sanfranciscensis, respectively, and that these LAB species also circulated in the corresponding bakery environment. Furthermore, AFLP fingerprinting demonstrated that sourdough and bakery environment isolates of these species were genetically indistinguishable. For more sensitive source-tracking, SYBR Green-based real-time PCR assays were developed using species-specific primers targeting the pheS gene of L. plantarum and L. sanfranciscensis, detected in air samples from D01 and D10, respectively. CONCLUSIONS: The results obtained in this study indicate that specific strains of LAB persist in artisan doughs over years and circulate in the bakery environment. Furthermore, the importance of air as a potential carrier of LAB in artisan bakery environments was demonstrated. SIGNIFICANCE AND IMPACT OF THE STUDY: PheS based real-time PCR can be used to detect, quantify and/or monitor specific LAB species (e.g. starter cultures) in sourdough and bakery environment samples. PMID- 19187145 TI - Methods for the recovery of a model virus from healthcare personal protective equipment. AB - AIMS: To develop methods for recovering a model virus (bacteriophage MS2) from healthcare personal protective equipment (PPE). METHODS AND RESULTS: Nine eluents were evaluated for recovery of infectious MS2 from PPE: 1.5% beef extract (BE) pH 7.5 with and without 0.1% Tween 80, 1.5% BE pH 9.0 with and without 0.1% Tween 80, 3% BE pH 7.5 with and without 0.1% Tween 80, 3% BE pH 9.0 with and without 0.1% Tween 80 and PBS with 0.1% Tween 80. Methods were applied to experimentally contaminated PPE. Elution followed by two-step enrichment assay could recover virus inputs as low as 1.5 log(10), and could recover >90% of inoculated virus from used items of experimentally contaminated PPE worn by human volunteers. CONCLUSIONS: BE was effective for recovering infectious viruses from a range of PPE materials. SIGNIFICANCE AND IMPACT OF THE STUDY: PPE plays a crucial role in interrupting transmission of infectious agents from patients to healthcare workers (HCWs). The fate of micro-organisms when PPE is removed and disposed of has important consequences for infection control. Methods described here can be used to conduct rigorous studies of viral survival and transfer on PPE for risk assessments in infection control and HCW protection. PMID- 19187146 TI - Adhesion of food-borne bacteria to stainless steel is reduced by food conditioning films. AB - AIMS: Preconditioning of stainless steel with aqueous cod muscle extract significantly impedes subsequent bacterial adhesion most likely due to repelling effects of fish tropomyosin. The purpose of this study was to determine if other food conditioning films decrease or enhance bacterial adhesion to stainless steel. METHODS AND RESULTS: Attachment of Pseudomonas fluorescens AH2 to stainless steel coated with water-soluble coatings of animal origin was significantly reduced as compared with noncoated stainless steel or stainless steel coated with laboratory substrate or extracts of plant origin. Coating with animal extracts also decreases adhesion of other food-relevant bacteria. The manipulation of adhesion was not attributable to growth inhibitory effects. Chemical analysis revealed that the stainless steels were covered by homogenous layers of adsorbed proteins. The presence of tropomyocin was indicated by appearance of proteins with similar molecular weight based in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, in several extracts that reduced adhesion but also extracts not containing this protein reduced bacterial adhesion, indicating that several molecular species may be involved in the phenomenon. CONCLUSIONS: It is a common perception that food materials facilitate bacterial adhesion to surfaces; however, this study demonstrates that aqueous coatings of food origin may actually reduce bacterial adhesion. SIGNIFICANCE AND IMPACT OF THE STUDY: Compounds from food extracts may potentially be used as nontoxic coatings to reduce bacterial attachment to inert surfaces. PMID- 19187147 TI - Effects of concentrated supernatants recovered from Lactobacillus plantarum on Escherichia coli growth and on the viability of a human promyelocytic cell line. AB - AIMS: The ability of concentrated supernatants from Lactobacillus plantarum to produce a disruption of plasma membrane in eukaryotic and prokaryotic cells has been examined. METHODS AND RESULTS: A strain of Lact. plantarum (tolerant to acid and bile salts and resistant to several antibiotics) was used. It inhibited the growth of pathogenic Escherichia coli and L. monocytogenes. Supernatants from Lact. plantarum were concentrated by centrifugation. Either E. coli or HL-60 cells (a human promyelocytic cell line) were treated in the presence of the concentrated supernatants. The effect of concentrated supernatants from Lact. plantarum on E. coli growth demonstrated a bacteriostatic activity and a loss of cell viability measured by sytox green staining. Concentrated supernatants were capable of disturbing plasma membrane in E. coli and of promoting a cytotoxic and lyctic action on HL-60 cells and on human erythrocytes, respectively. CONCLUSIONS: These results suggest that Lact. plantarum release an effective compound responsible for an important effect in the disruption of E. coli plasma membrane and for a cytototoxic activity on promyelocytic leukaemia cells. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first in vitro study about the antimicrobial and biological activities of concentrated supernatants from Lact. plantarum. PMID- 19187148 TI - Rapid and simple detection of Clostridium botulinum types A and B by loop mediated isothermal amplification. AB - AIMS: To develop a convenient and rapid detection method for toxigenic Clostridium botulinum types A and B using a loop-mediated isothermal amplification (LAMP) method. METHODS AND RESULTS: The LAMP primer sets for the type A or B botulinum neurotoxin gene, BoNT/A or BoNT/B, were designed. To determine the specificity of the LAMP assay, a total of 14 C. botulinum strains and 17 other Clostridium strains were tested. The assays for the BoNT/A or BoNT/B gene detected only type A or B C. botulinum strains, respectively, but not other types of C. botulinum or strains of other Clostridium species. Using purified chromosomal DNA, the sensitivity of LAMP for the BoNT/A or BoNT/B gene was 1 pg or 10 pg of DNA per assay, respectively. The assay times needed to detect 1 ng of DNA were only 23 and 22 min for types A and B, respectively. In food samples, the detection limit per reaction was one cell for type A and 10 cells for type B. CONCLUSIONS: The LAMP is a sensitive, specific and rapid detection method for C. botulinum types A and B. SIGNIFICANCE AND IMPACT OF THE STUDY: The LAMP assay would be useful for detection of C. botulinum in environmental samples. PMID- 19187149 TI - Assessment of anti-oxidant activity of plant extracts using microbial test systems. AB - AIMS: To evaluate the anti-oxidant properties of extracts from 20 medicinal herbs growing in western Siberia using microbial test systems and different in vitro methods. METHODS AND RESULTS: In vivo anti-oxidant activity of extracts was evaluated for their capacity to protect bacteria, Escherichia coli, against bacteriostatic and bactericidal effects of H(2)O(2) and menadione, and action on anti-oxidant gene expression. In vitro anti-oxidant activity has been examined by a number of methods including: the 1,1-diphenyl-2-picrylhydrazyl radical (DPPH(*))-scavenging assay, chelating activity and capacity to protect plasmid DNA against oxidative damage. In addition, total polyphenol content was determined. The extracts of Fragaria vesca, Rosa majalis, Pentaphylloides fruticosa, Alchemilla vulgaris and Pulmonaria mollis possessed the highest levels of anti-oxidant activity in vivo and in vitro. The protective properties were more closely related to the DPPH(*) radical-scavenging activity, tannin content and action on anti-oxidant gene expression than to other parameters. CONCLUSION: The extracts of medicinal plants may have anti-oxidant effects on bacteria simultaneously through several different pathways, including direct inhibition of reactive oxygen species, iron chelation and anti-oxidant genes induction. SIGNIFICANCE AND IMPACT OF THE STUDY: Using microbial test systems, we revealed herbs that may be used as potential sources of natural anti-oxidants. PMID- 19187150 TI - Effects of temperature on metal tolerance and the accumulation of Zn and Pb by metal-tolerant fungi isolated from urban runoff treatment wetlands. AB - AIMS: To investigate the ability of two fungi to accumulate Zn and Pb, the effect of temperature on their metal tolerance and possible mechanisms involved in metal accumulation. METHODS AND RESULTS: Beauveria bassiana and Rhodotorula mucilaginosa isolated from constructed wetlands receiving urban runoff were grown in modified glycerol asparagine medium containing elevated levels of Zn and Pb at 30 degrees C. Beauveria bassiana accumulated up to 0.64% of available Zn and 8.44% of Pb. The corresponding values for R. mucilaginosa were up to 2.05% for Zn and 16.55% for Pb. Radial growth of colonies grown at 4 degrees and 30 degrees C on agar containing Zn or Pb indicated that metal tolerance was not seriously affected by a decrease in temperature. Transmission electron microscopy and emission dispersion x-ray spectrophotometry suggested that the mechanism of resistance in B. bassiana may be associated with the precipitation of Pb (possibly in the form of oxalates). CONCLUSION: The processes of biosorption could potentially occur throughout the year with both living and dead cells able to accumulate metals. SIGNIFICANCE AND IMPACT OF THE STUDY: Identified precipitation processes could be an important mechanism in metal removal in wetland substrates serving as long-term storage sinks. PMID- 19187152 TI - Brevibacillus laterosporus MTCC 2298: a potential azo dye degrader. AB - AIMS: To evaluate the potential of Brevibacillus laterosporus MTCC 2298 for the decolourization of different textile azo dyes including methyl red, mechanism of biotransformation and the toxicity of products. METHODS AND RESULTS: Brevibacillus laterosporus showed decolourization of thirteen different azo dyes including methyl red. Decolourization of methyl red was faster (93% within 12 h) under static condition at the concentration 0.2 g l(-1). Induction in the activities of lignin peroxidase, laccase, aminopyrine N-demethylase, NADH-DCIP reductase and malachite green reductase was observed in the cells obtained after decolourization. Fourier transform infra-red spectral analysis of products indicated conversion of methyl red into secondary aryl amines and nitrosamines, which further transformed into the aromatic nitro compounds. Gas chromatography mass spectroscopy analysis suggested conversion of methyl red into high molecular weight complex derivatives. The heterocyclic substituted aryl amine (m/z 281), p (N,N di formyl)-substituted para-di amino benzene derivative (m/z 355) and p-di amino benzene derivative (m/z 282) are the mainly elected biotransformation products. Microbial and phytotoxicity studies suggested nontoxic nature of the biotransformation products. CONCLUSIONS: Brevibacillus laterosporus has potential for the decolourization of different textile azo dyes. SIGNIFICANCE AND IMPACT OF THE STUDY: Brevibacillus laterosporus decolourized different azo dyes including methyl red and can be utilized for textile dye decolourization. PMID- 19187151 TI - A strict anaerobic extreme thermophilic hydrogen-producing culture enriched from digested household waste. AB - AIMS: The aim of this study was to enrich, characterize and identify strict anaerobic extreme thermophilic hydrogen (H(2)) producers from digested household solid wastes. METHODS AND RESULTS: A strict anaerobic extreme thermophilic H(2) producing bacterial culture was enriched from a lab-scale digester treating household wastes at 70 degrees C. The enriched mixed culture consisted of two rod shaped bacterial members growing at an optimal temperature of 80 degrees C and an optimal pH 8.1. The culture was able to utilize glucose, galactose, mannose, xylose, arabinose, maltose, sucrose, pyruvate and glycerol as carbon sources. Growth on glucose produced acetate, H(2) and carbon dioxide. Maximal H(2) production rate on glucose was 1.1 mmol l(-1) h(-1) with a maximum H(2) yield of 1.9 mole H(2) per mole glucose. 16S ribosomal DNA clone library analyses showed that the culture members were phylogenetically affiliated to the genera Bacillus and Clostridium. Relative abundance of the culture members, assessed by fluorescence in situ hybridization, were 87 +/- 5% and 13 +/- 5% for Bacillus and Clostridium, respectively. CONCLUSIONS: An extreme thermophilic, strict anaerobic, mixed microbial culture with H(2)-producing potential was enriched from digested household wastes. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provided a culture with a potential to be applied in reactor systems for extreme thermophilic H(2) production from complex organic wastes. PMID- 19187153 TI - Cell wall component and mycotoxin moieties involved in the binding of fumonisin B1 and B2 by lactic acid bacteria. AB - AIMS: The ability of lactic acid bacteria (LAB) to bind fumonisins B1 and B2 (FB1, FB2) in fermented foods and feeds and in the gastrointestinal tract could contribute to decrease their bioavailability and toxic effects on farm animals and humans. The aim of this work was to identify the bacterial cell wall component(s) and the functional group(s) of FB involved in the LAB-FB interaction. METHODS AND RESULTS: The effect of physicochemical, enzymatic and genetic treatments of bacteria and the removal/inactivation of the functional groups of FB on toxin binding were evaluated. Treatments affecting the bacterial wall polysaccharides, lipids and proteins increased binding, while those degrading peptidoglycan (PG) partially decreased it. In addition, purified PG from Gram-positive bacteria bound FB in a manner analogue to that of intact LAB. For FB, tricarballylic acid (TCA) chains play a significant role in binding as hydrolysed FB had less affinity for LAB. CONCLUSIONS: Peptidoglycan and TCA are important components of LAB and FB, respectively, involved in the binding interaction. SIGNIFICANCE AND IMPACT OF THE STUDY: Lactic acid bacteria binding efficiency seems related to the peptide moiety structure of the PG. This information can be used to select probiotics with increased FB binding efficiency. PMID- 19187154 TI - The structure of the bacterial and archaeal community in a biogas digester as revealed by denaturing gradient gel electrophoresis and 16S rDNA sequencing analysis. AB - AIMS: To identify the bacterial and archaeal composition in a mesophilic biogas digester treating pig manure and to compare the consistency of two 16S rDNA-based methods to investigate the microbial structure. METHODS AND RESULTS: Sixty-nine bacterial operational taxonomic units (OTU) and 25 archaeal OTU were identified by sequencing two 16S rDNA clone libraries. Most bacterial OTU were identified as phyla of Firmicutes (47.2% of total clones), Bacteroides (35.4%) and Spirochaetes (13.2%). Methanoculleus bourgensis (29.0%), Methanosarcina barkeri (27.4%) and Methanospirillum hungatei (10.8%) were the dominant methanogens. Only 9% of bacterial and 20% of archaeal OTU matched cultured isolates at a similarity index of >or=97%. About 78% of the dominant bacterial (with abundance >3%) and 83% of archaeal OTU were recovered from the denaturing gradient gel electrophoresis (DGGE) bands of V3 regions in 16S rDNAs. CONCLUSIONS: In the digester, most bacterial and archaeal species were uncultured; bacteria belonging to Firmicutes, Bacteroides and Spirochaetes seem to take charge of cellulolysis, proteolysis, acidogenesis, sulfur-reducing and homoacetogenesis; the most methanogens were typical hydrogenotrophic or hydrogenotrophic/aceticlastic; DGGE profiles reflected the dominant microbiota. SIGNIFICANCE AND IMPACT OF THE STUDY: This study gave a first insight of the overall microbial structure in a rural biogas digester and also indicated DGGE was useful in displaying its dominant microbiota. PMID- 19187155 TI - Survival of Mycobacterium avium attached to polyethylene terephtalate (PET) water bottles. AB - AIMS: The main objective of our study was to assess the persistence of Mycobacterium avium in an oligotrophic environment such as bottled groundwater. METHODS AND RESULTS: Filtered groundwater samples were spiked with washed Myco. avium suspension and stored in dark and under static conditions, at 20 degrees C, for 3 months in 500 ml PET bottles. The loss of Myco. avium cultivability was slow in water. On the contrary, after a 3-month storage at 20 degrees C, growth of attached cells was observed and cell adhesiveness to the PET wall increased with time. It could probably be because of the presence of an extracellular matrix. CONCLUSIONS: This study has shown the great stability of Myco. avium in bulk water as well as their adhesiveness and their growth on a PET bottle wall in an oligotrophic environment. SIGNIFICANCE AND IMPACT OF THE STUDY: Slowly growing mycobacteria are well adapted to oligotrophic environments such as groundwater. As they stick very well to surfaces, they could be used for determining the efficiency of the cleaning of contaminated surfaces. PMID- 19187156 TI - Analysis of dye binding by and membrane potential in spores of Bacillus species. AB - AIMS: To determine roles of coats in staining Bacillus subtilis spores, and whether spores have membrane potential. METHODS AND RESULTS: Staining by four dyes and autofluorescence of B. subtilis spores that lack some (cotE, gerE) or most (cotE gerE) coat protein was measured. Wild-type, cotE and gerE spores autofluorescenced and bound dyes, but cotE gerE spores did not autofluorescence and were stained only by two dyes. A membrane potential-sensitive dye DiOC6(3) bound to dormant Bacillus megaterium and B. subtilis spores. While this binding was abolished by the protonophore FCCP, DiOC6(3) bound to heat-killed spores, but not to dormant B. subtilis cotE gerE spores. However, DiOC6(3) bound well to all germinated spores. CONCLUSIONS: The autofluorescence of dormant B. subtilis spores and the binding of some dyes are due to the coat. There is no membrane potential in dormant Bacillus spores, although membrane potential is generated when spores germinate. SIGNIFICANCE AND IMPACT OF THE STUDY: The elimination of the autofluorescence of B. subtilis spores may allow assessment of the location of low abundance spore proteins using fluorescent reporter technology. The dormant spore's lack of membrane potential may allow tests of spore viability by assessing membrane potential in germinating spores. PMID- 19187158 TI - Diversity of hydrocarbon-degrading Klebsiella strains isolated from hydrocarbon contaminated estuaries. AB - AIMS: To investigate the diversity and the catabolic capacity of oil-degrading Klebsiella strains isolated from hydrocarbon-contaminated sediments in Santos-Sao Vicente estuary systems in Brazil. METHODS AND RESULTS: Klebsiella strains obtained from the estuary were characterized using 16S rRNA gene sequencing and BOX-PCR patterns, testing their catabolic capacity to degrade toluene, xylene, naphthalene and nonane, and identifying the catabolic genes present in the oil degrading strains. Results show that Klebsiella strains were widespread in the estuary. Twenty-one isolates from the Klebsiella genus were obtained; 14 had unique BOX patterns and were further investigated. Among four distinct catabolic genes tested (todC1, ndoB, xylE and alkB1), only the todC1 gene could be amplified in two Klebsiella strains. The biodegradation assay showed that most of the strains had the ability to degrade all of the tested hydrocarbons; however, the strains displayed different efficiencies. CONCLUSIONS: The oil-degrading Klebsiella isolates obtained from the estuary were closely related to Klebsiella pneumoniae and Klebsiella ornithinolytica. The isolates demonstrated a substantial degree of catabolic plasticity for hydrocarbon degradation. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this study show that several strains from the Klebsiella genus are able to degrade diverse hydrocarbon compounds. These findings indicate that Klebsiella spp. can be an important part of the oil-degrading microbial community in estuarine areas exposed to sewage. PMID- 19187159 TI - Real-time PCR detection and quantification of fish probiotic Phaeobacter strain 27-4 and fish pathogenic Vibrio in microalgae, rotifer, Artemia and first feeding turbot (Psetta maxima) larvae. AB - AIMS: To develop a SYBR Green quantitative real-time PCR protocol enabling detection and quantification of a fish probiotic and two turbot pathogenic Vibrio spp. in microcosms. METHODS AND RESULTS: Phaeobacter 27-4, Vibrio anguillarum 90 11-287 and Vibrio splendidus DMC-1 were quantified as pure and mixed cultures and in presence of microalgae (Isochrysis galbana), rotifers (Brachionus plicatilis), Artemia nauplii or turbot (Psetta maxima) larvae by real-time PCR based on primers directed at genetic loci coding for antagonistic and virulence-related functions respectively. The optimized protocol was used to study bioencapsulation and maintenance of the probiont and pathogens in rotifers and for the detection and quantification of Phaeobacter and V. anguillarum in turbot larvae fed rotifers loaded with the different bacteria in a challenge trial. CONCLUSIONS: Our real-time PCR protocol is reproducible and specific. The method requires separate standard curve for each host organism and can be used to detect and quantify probiotic Phaeobacter and pathogenic Vibrio bioencapsulated in rotifers and in turbot larvae. SIGNIFICANCE AND IMPACT OF THE STUDY: Our method allows monitoring and quantification of a turbot larvae probiotic bacteria and turbot pathogenic vibrios in in vivo trials and will be useful tools for detecting the bacteria in industrial rearing units. PMID- 19187160 TI - Elevation of Francisella philomiragia subsp. noatunensis Mikalsen et al. (2007) to Francisella noatunensis comb. nov. [syn. Francisella piscicida Ottem et al. (2008) syn. nov.] and characterization of Francisella noatunensis subsp. orientalis subsp. nov., two important fish pathogens. AB - AIMS: This study was conducted to clarify the taxonomic status of Francisella sp. strain Ehime-1, a fish pathogen, in relation to the fish pathogens F. piscicida and F. philomiragia subsp. noatunensis and to F. philomiragia subsp. philomiragia. METHODS AND RESULTS: Francisella sp. Ehime-1 was compared to F. piscicida, F. philomiragia subsp. noatunensis and several F. philomiragia subsp. philomiragia isolates through sequencing of the 16S rRNA-gene and several house keeping genes and determination of biochemical and phenotypic properties. Results show that F. piscicida is indistinguishable from F. philomiragia subsp. noatunensis by sequence and phenotypic traits. Francisella sp. Ehime-1 and F. philomiragia subsp. noatunensis are clearly separated from F. philomiragia. Francisella sp. Ehime-1 is biochemically, phenotypically and genetically different from F. philomiragia subsp. noatunensis (=F. piscicida), but DNA-DNA hybridization does not clearly support establishment as a separate species (level of relatedness 64% and 73.4%, mean 68.7%). CONCLUSIONS: We propose to elevate F. philomiragia subsp. noatunensis to species rank as F. noatunensis comb. nov., while F. piscicida is considered a heterotypic synonym of F. noatunensis comb. nov. Evidence suggests that Francisella sp. Ehime-1 represents a novel subspecies of F. noatunensis, for which the name F. noatunensis subsp. orientalis subsp. nov. is proposed (=DSM21254(T), = LMG24544(T)). SIGNIFICANCE AND IMPACT OF THE STUDY: This study contributes to the taxonomy and characteristics of fish pathogenic Francisella spp. PMID- 19187161 TI - Localization of norovirus and poliovirus in Pacific oysters. AB - AIMS: To examine the uptake and tissue distribution of norovirus (NoV) and poliovirus (PV) experimentally bioaccumulated in feeding Pacific oysters (Crassostrea gigas). METHODS AND RESULTS: Pacific oysters were allowed to bioaccumulated either PV or NoV under tidally synchronized feeding conditions in laboratory tanks. Oysters were then either fixed and paraffin wax embedded prior to localizing virus within tissues by immunohistochemistry (IHC), or they were dissected into digestive tract (stomach, intestine and digestive diverticula), gill and labial palp tissues, and the viral load determined by quantitative RT PCR. Both PV and NoV immunoreactivities were predominantly found in the lumen and within cells of the digestive tract tissues; however, PV was also found within cells of nondigestive tract tissues, and in the gills and labial palp. Quantitative RT-PCR of tissue extracts corroborate the immunohistochemical data in that the major site for virus localization is the gut, but significant amounts of viral RNA were identified in the gills and labial palp. CONCLUSIONS: The human enteric viruses, PV and NoV, are readily bioaccumulated by feeding Pacific oysters and that some of the virus is internalized within cells of both digestive and nondigestive tissues. SIGNIFICANCE AND IMPACT OF THE STUDY: Oysters that have been virally contaminated even after depuration (cleaning) in uncontaminated seawater could pose a human health risk if consumed. PMID- 19187162 TI - Seasonal variation of Shiga toxin-encoding genes (stx) and detection of E. coli O157 in dairy cattle from Argentina. AB - AIMS: To study the seasonal variation of Shiga toxin-encoding genes (stx) and to investigate the presence of Shiga toxin-producing Escherichia coli (STEC) O157 in cattle belonging to five dairy farms from Argentina. METHODS AND RESULTS: Rectal swab samples were collected from 360 dairy cows in each season and 115 and 137 calves in autumn and in spring, respectively. The stx were investigated by multiplex PCR and it was used as the indicator for STEC. Samples positives for stx were tested by PCR for eae-gamma1 of E. coli O157 and then subjected to IMS (immunomagnetic separation). In positive animals significant differences in the prevalence of stx between warm and cold seasons were detected. In warm seasons, stx1 + stx2 increased and stx1 decreased, independently of the animal category. The prevalence of STEC O157 in cows and calves were 0.2% and 0.8%, respectively. CONCLUSIONS: This work provides new data about the occurrence of stx and STEC O157 in dairy herds from Argentina and suggests a relationship between the type of stx and season of year. SIGNIFICANCE AND IMPACT OF STUDY: The detection of STEC O157 and the seasonality of stx and its types provide an opportunity to improve control strategies designed to prevent contamination of food products and transmission animal-person. PMID- 19187163 TI - Problems faced by relatives caring for cancer patients at home. AB - The care of patients suffering from advanced cancer is not limited in the hospital setting. It continues at home where the burden of care is borne by specific individuals. The aim of the present study was to survey and record the various problems faced by those who care for cancer patients at home. The study was conducted in our hospital during the summer of 2007. All participants completed, during a personal interview, a questionnaire which covered pathologic, social, psychological, spiritual/religious and financial problems. Seventy-six carers returned fully completed questionnaires. The most frequent problems reported were: anxiety regarding the patient's future (61.8%), troublesome symptoms such as pain (54%), increased economic burden-financial difficulty (51.3%), problems with patient's feeding (50%), unhappiness or depression (48,7%), emotional upset (47.4%), worsening of the patient's behaviour and personality (38.2%), difficulty of establishing a positive attitude regarding their current status (34.2%), transport to hospital (32.9%), assistance from the wider family circle (25%). Taking care of cancer patients at home creates several problems among carers. Many of them remain undetected. The acknowledgement and recognition of these problems by health-care professionals might contribute to finding solutions in order to assist the difficult task of these individuals. PMID- 19187164 TI - Husserl and Heidegger: exploring the disparity. AB - Introduced as an alternative to empirical science, phenomenology offers nursing an insightful means for understanding nursing phenomena specifically in relation to lived experiences. However, not all phenomenologies were created equal, a point which has left many a nursing researcher not only confused. Furthermore, this confusion might result in the choosing of a philosophical framework that is neither cognizant with the research question nor the epistemological lens through which the researcher operates. Drawing on common nursing examples to illustrate concepts, the authors closely examine and debate the disparities between Husserl's transcendental phenomenology and Heidegger's hermeneutic approach to phenomenology. The aim of the article is to demystify the dense language used and present the fundamental beliefs of each philosopher in a format that is accessible to novice phenomenologists. PMID- 19187165 TI - Systematic review of the effectiveness of primary care nursing. AB - This paper reports on a systematic review that sought to answer the research question: What is the impact of the primary and community care nurse on patient health outcomes compared with usual doctor-led care in primary care settings? A range of pertinent text-words with medical subject headings were combined and electronic databases were searched. Because of the volume of published articles, the search was restricted to studies with high-level evidence. Overall, 31 relevant studies were identified and included in the review. We found modest international evidence that nurses in primary care settings can provide effective care and achieve positive health outcomes for patients similar to that provided by doctors. Nurses are effective in care management and achieve good patient compliance. Nurses are also effective in a more diverse range of roles including chronic disease management, illness prevention and health promotion. Nevertheless, there is insufficient evidence about primary care nurses' roles and impact on patient health outcomes. PMID- 19187166 TI - Medication non-adherence--exploring the conceptual mire. AB - It is well established that pharmacotherapy is fundamental to disease and symptom management. Adherence to medication regimens is therefore essential for improved patient health outcomes. In reality, however, many patients experience difficulty with medication taking resulting in suboptimal adherence. Given the consequences of this pervasive problem, non-adherence is increasingly recognized as one of the leading challenges that professionals face in contemporary health care. As health care professionals, nurses have an important role in combating this problem. This paper therefore examines the literature surrounding medication adherence for the purposes of enhancing professional knowledge and practice in this area. Specifically, the impact of poor adherence from the patient's, health-care professional and health-care system perspective is detailed to highlight the significance of this issue. The underlying philosophical attributes of adherence and the interchangeable concepts of compliance and concordance are explored, with the implications of the meanings attributed to these concepts for nursing practice and research considered. PMID- 19187167 TI - Breastfeeding practices among Jordanian women. AB - The purpose of the study was to explore Jordanian women's breastfeeding beliefs and practices including exclusive breastfeeding. A descriptive cross-sectional design with a convenience sample of 200 Jordanian mothers was used. The majority of mothers were muliparous and were recruited from primary health-care centres within 6 weeks of a normal vaginal birth or an instrumental delivery. Eligible women, who met the inclusion criteria, were invited to participate in the study. A sociodemographic data form and a 14-item questionnaire concerning different aspects of breastfeeding beliefs and practices were developed for self administration. This study indicated high early initiation of breastfeeding. Most mothers gave supplements other than breastfeeding, including water without knowing that this supplementation could affect exclusive breastfeeding or the continuation of breastfeeding. Finding of this study shed some light on the current breastfeeding practices including exclusive breastfeeding among Jordanian women. Women need to be better educated about breastfeeding. Therefore, more efforts and resources should be put into providing opportunities for education to discuss breastfeeding during antenatal care. This Jordanian study could be relevant to Arabic women in the West, because cultural beliefs and practices are likely to be part of immigrant woman's perceptions about breastfeeding practices. PMID- 19187168 TI - Challenges for professional care of advanced dementia. AB - Qualitative methodology based on action research identified challenges when caring for persons with advanced dementia, as perceived by key professional providers. Data collection was via five focus groups (total n = 24) and 20 follow up individual interviews. Participants included palliative care, aged care and dementia specialist nurses, medical specialists from an area health service, residential aged care staff and general medical practitioners. Responses emphasized the need for improved knowledge and skills, and clearer policy. Concerns included accurate assessment, especially of pain, owing to the inability of people with advanced dementia to communicate their symptoms. Assessment, managing physical and behavioural symptoms, and communicating with family presented as further challenges. Conclusions are that the need for a palliative approach to care in advanced dementia should be recognized. Aged care staff can deliver palliative care to people with advanced dementia only if the staff receive relevant education and training beyond their generalist competencies. PMID- 19187169 TI - 'Knowing what makes them tick': motivating cognitively impaired older adults to participate in restorative care. AB - Nursing home residents with dementia represent a majority of the most functionally impaired individuals residing in nursing homes. Although many perceive this population as having little restorative potential, maintaining resident functional abilities for as long as possible helps to optimize quality of life and decrease caregiver burden. This study used a qualitative design with a focus group methodology to explore facilitators and barriers to engaging cognitively impaired residents in functional activities and exercise. A purposive sample of seven geriatric nursing assistants who were experts in dementia care participated in the study. Twenty-seven codes were reduced to three themes: (i) knowing what makes them tick and move; (ii) teamwork and utilizing resources; and (iii) barriers to restorative care. The study findings were used to revise the Restorative Care for the Cognitively Impaired Intervention and could direct future implementation of programmes in nursing home settings. PMID- 19187170 TI - Initiating insulin therapy with, or switching existing insulin therapy to, biphasic insulin aspart 30/70 (NovoMix 30) in routine care: safety and effectiveness in patients with type 2 diabetes in the IMPROVE observational study. AB - AIMS: The IMPROVE observational study evaluated the safety profile and effectiveness of biphasic insulin aspart 30/70 (BIAsp 30) in patients with type 2 diabetes in routine practice in 11 countries. METHODS: Patients who initiated insulin therapy with, or switched existing insulin therapy to, BIAsp 30 in routine care were eligible for this 26-week, non-interventional observational study. Data on adverse events, hypoglycaemia and glycaemic parameters were obtained from patients' diaries and medical notes. Questionnaire-based patient treatment satisfaction was also measured. We report global results and, uniquely for a diabetes observational study, country-specific data. RESULTS: A total of 52,419 patients were enrolled from three prestudy treatment groups: no pharmaceutical therapy (n = 8966, diabetes duration 2.0 years, baseline HbA1c 9.9%), oral antidiabetic drugs (OADs) only (n = 33,797, diabetes duration 7.4 years, baseline HbA1c 9.2%) and insulin +/- OADs (n = 9568, diabetes duration 10.4 years, baseline HbA1c 9.3%). At final visit, HbA1c, fasting and postprandial blood glucose were significantly reduced from baseline in all subgroups (no pharmaceutical therapy: -3.1%, -5.9 and -9.0 mmol/l, respectively; OADs-only: 2.1%, -4.1 and -6.1 mmol/l; insulin +/- OADs: -2.0%, -3.3 and -5.1 mmol/l). Major hypoglycaemia rates decreased in all subgroups; minor hypoglycaemia increased in the insulin-naive groups. There was no mean weight gain across subgroups. Across all countries, glycaemic parameters and major hypoglycaemia were reduced; weight increases were seen in some countries. Treatment satisfaction increased in all subgroups and countries following BIAsp 30 therapy. CONCLUSIONS: Initiating insulin with, or switching insulin therapy to, BIAsp 30 in routine care resulted in improved glycaemic control, reduced major hypoglycaemia and greater treatment satisfaction. PMID- 19187171 TI - Objective amount of limb fat in HIV-infected subjects with subjective diagnosis of lipoatrophy. AB - BACKGROUND: The relationship between the subjective diagnosis of lipoatrophy and the objective amount of limb fat loss in HIV-infected adults is unclear. METHODS: Using Medline, we identified published articles reporting the amount of arm, leg or limb fat measured by dual X-ray absorptiometry in HIV-infected patients with moderate-severe lipoatrophy and in healthy non-HIV-infected adults. We calculated the relative content of fat in the limbs, arms and legs of lipoatrophic patients with regard to the weighted arithmetic means of those fat values in healthy controls. RESULTS: We found 799 patients from 10 articles, and 73 healthy controls from two articles. Limb fat ranged from 2.6 to 4.4 kg in patients, and from 7.1 to 7.2 kg in controls. Both patients and controls were almost exclusively men, of white race, and in their forties. Weighted arithmetic means of arm, leg and limb fat in HIV-infected patients with clinically evident lipoatrophy were 1.0, 2.1 and 3.1 kg, respectively (48, 41 and 43% relative to healthy non-HIV-infected males, respectively). CONCLUSIONS: The diagnosis of lipoatrophy was highly correlated with the amount of limb fat, irrespective of the investigators. HIV-infected men with clinically evident lipoatrophy had a limb fat loss of >50% compared with non-HIV-infected healthy males. PMID- 19187172 TI - Neurocognitive screening tools in HIV/AIDS: comparative performance among patients exposed to antiretroviral therapy. AB - OBJECTIVE: The aim of the study was to compare the performance of several bedside neuropsychological tools for detection of HIV-associated neurocognitive disorder (HAND) in antiretroviral drug-exposed persons. METHODS: We analysed the relative performance of the HIV Dementia Scale (HDS), International HIV Dementia Scale (IHDS) and the Mini-Mental Status Exam (MMSE) together with neuropsychological tests (Symbol-Digit, Grooved Pegboard and Trail Making) in HIV-1-seronegative subjects (HIV-; n=13) and in HIV-1-seropositive subjects with HAND (HIV+HAND; n=13) and other neurological disorders (HIV+OND; n=20). RESULTS: Established neuropsychological tests consistently showed significantly poorer performance by HIV+HAND subjects compared with the other two groups. Similarly, the mean HDS and IHDS scores were lower in the HIV+HAND group compared with the other two groups (P<0.005) while the mean MMSE score did not show significant differences between the HIV+HAND and HIV+OND groups. Receiver operator characteristics curves generated from these data using predefined cut-off scores revealed that the HDS, IHDS and MMSE displayed corresponding area under the curve values of 0.82, 0.74 and 0.48, respectively (P<0.006). CONCLUSIONS: The present findings indicate that the MMSE is a weak tool for diagnosing HAND in this group of patients but the HDS and IHDS demonstrate better efficiencies, although cut-off values for the HDS require reassessment in the era of effective antiretroviral therapy. PMID- 19187173 TI - Vertically acquired HIV diagnosed in adolescence and early adulthood in the United Kingdom and Ireland: findings from national surveillance. AB - OBJECTIVE: The aim of the study was to describe the characteristics of young people with vertically acquired HIV diagnosed aged > or =13 years. METHODS: A retrospective review of HIV diagnoses reported to well-established national paediatric and adult HIV surveillance systems in the United Kingdom/Ireland was conducted. RESULTS: Forty-two young people with vertically acquired HIV diagnosed aged > or =13 years were identified; 23 (55%) were female, 40 (95%) were black African and 36 (86%) were born in sub-Saharan Africa. The median age at HIV diagnosis was 14 years (range, 13-20 years). Half of the patients presented with symptoms; the remainder were screened for HIV following diagnosis of a relative. The median CD4 count at diagnosis was 210 cells/microL (range, 0-689 cells/microL), 12 patients (29%) were diagnosed with AIDS at HIV diagnosis or subsequently, and 34 (81%) started combination antiretroviral therapy (ART), most (31 of 34) within a year of diagnosis. CONCLUSION: A small number of young people with vertically acquired HIV survive childhood without ART and are diagnosed at age > or =13 years in the United Kingdom/Ireland. Half of the patients were asymptomatic, highlighting the importance of considering HIV testing for all offspring of HIV-infected women, regardless of age or symptoms. Increased awareness among clinicians and parents is required to reduce delayed presentation with advanced disease and to avoid onward transmission as these young people become sexually active. PMID- 19187174 TI - Comparison of glomerular filtration rate estimates vs. 24-h creatinine clearance in HIV-positive patients. AB - BACKGROUND: Guidelines for kidney function monitoring and antiretroviral drug dosing are available and respectively refer to glomerular filtration rate and creatinine clearance (CrCl). OBJECTIVE: The aim of the study was to compare kidney function estimates vs. measured 24-h CrCl in HIV-infected subjects. METHODS: A cross-sectional design was used, with comparison of Cockcroft-Gault (CG), original and simplified modification of diet in renal disease (MDRD) equations vs. measured 24-h CrCl. Subjects were HIV-infected, 18-70 years old, without pre-existing kidney disease. RESULTS: Results are presented as mean (+/ standard deviation), unless otherwise stated. The study population consisted of 90 patients, of whom 71% were male, with a mean age of 45 years (+/-6.5 years). At the time of evaluation, the mean body mass index was 23 (+/-3.3); mean serum creatinine was 0.91 mg/dL (+/-0.2 mg/dL); and mean blood urea nitrogen (BUN) was 34.7 mg/dL (+/-10.6 mg/dL). Differences between paired methods were all significant (P<0.00001), except between CG and simplified MDRD (P=0.21; Pearson r=0.81). In univariate analysis, male gender, CD4 nadir, hepatitis B virus coinfection, BUN and current CD4 cell count showed a significant positive correlation (P<0.2) with the difference between measured 24-h CrCl and either CG or simplified MDRD estimates. In multivariate analysis, only BUN showed a significant positive correlation (P<0.05). CONCLUSIONS: Estimates were lower than the measurements of 24-h CrCl. Original MDRD estimates were lower than those with other equations. CG and simplified MDRD estimates showed a satisfactory correlation. PMID- 19187175 TI - Whither thoracic pathology? PMID- 19187176 TI - Reporting lung cancer pathology specimens. Impact of the anticipated 7th Edition TNM classification based on recommendations of the IASLC Staging Committee. AB - Led by the International Association for the Study of Lung Cancer (IASLC), there are currently several major international collaborative projects underway that will have a significant impact on the future reporting of lung cancer pathology. In particular, the IASLC Staging Committee has just completed an analysis of >100,000 lung cancer cases, providing the basis for proposed revisions of the current TNM staging classification. The purpose of this review is not to provide a comprehensive document on recommendations for specimen processing, but rather to discuss how the anticipated changes in the 7th edition TNM will impact on specimen processing, specifically looking at tumour size, how to deal with multiple tumours and visceral pleural invasion. TNM staging of carcinoid tumours and small cell carcinoma is also discussed. PMID- 19187177 TI - Pulmonary adenocarcinomas: classification and reporting. AB - Pulmonary adenocarcinoma is the most common, and the most diverse form of primary lung carcinoma. The histological complexity of these tumours poses problems for pathologists. The current WHO classification of pulmonary adenocarcinoma does not adequately address a number of clinically relevant, biological factors. The accurate diagnosis of adenocarcinoma on small biopsy specimens, accounting for most diagnoses of this disease, is challenged by the absence of tumour architecture in most samples. Tumours showing a pure bronchioloalveolar (BAC) pattern are now best regarded as adenocarcinoma-in-situ; yet invasive adenocarcinomas may also show elements with the BAC pattern, dictating a better prognosis but biologically not necessarily in-situ disease. Multifocal BAC pattern adenocarcinoma still poses considerable conceptual and diagnostic problems. In small tumours the papillary pattern, especially when micropapillary, confers a poor prognosis but this is not reflected in larger tumours. In early tumours of predominantly BAC (in-situ) pattern, the identification of invasion is particularly difficult, yet minor degrees of infiltration seem not to degrade prognosis. It may therefore be possible to define a minimally invasive category of adenocarcinoma. Consequently, there are a number of issues to consider when reporting this tumour type, depending on the nature of the diagnostic specimen. The rapid emergence of chemotherapeutic agents with histology-specific efficacy will increase the need for more accurate and specific diagnosis of adenocarcinoma on small samples. Immunohistochemistry may help suggest this diagnosis when the features are non-specific but immunohistochemical findings are not diagnostic of this form of lung cancer. The emerging clinical and prognostic relevance of a number of histological features in these complex tumours strengthens the argument in favour of including quantitative detail of pattern sub-types in reports on resected tumours. PMID- 19187178 TI - The challenge of classifying poorly differentiated tumours in the lung. AB - The lung is a common site for primary carcinomas as well as a wide range of metastatic carcinomas and other rarer primary and secondary tumours that clinically and radiologically may mimic lung cancer. The World Health Organization classification of lung tumours is based on the morphological appearances in resected specimens. This provides a widely applicable system for classification, which, although complex, can be applied universally. The vast majority of patients with lung cancer, however, do not have their tumours resected, and the application of the same classification system to small biopsy specimens is problematic due in large part to sampling. Recognition of this has led to the widespread use of the term 'non-small cell carcinoma'. Future developments in our understanding of lung cancer and the development of novel targeted therapies may mean that in the future classification will become based more on the molecular features of tumours that predict response to therapies and prognosis rather than simple morphology. The predictive validity of such an approach using small biopsy or cytology specimens, however, still requires to be established. The problem of tumour heterogeneity, which is problematic for morphological classification, may also pose similar challenges for molecular approaches. PMID- 19187179 TI - Pulmonary preneoplasia--sequential molecular carcinogenetic events. AB - Bronchial and bronchioloalveolar carcinogenesis is a multicentric and multistep process, leading to a sequential accumulation of molecular and genetic abnormalities, mainly due to exposure to tobacco carcinogens. Concomitantly, a series of morphological alterations of normal bronchial or bronchioloalveolar epithelium occur, resulting in preneoplastic and then neoplastic lesions. The three pulmonary preneoplastic changes recognized to date in the lung include bronchial squamous dysplasia and in situ carcinoma, preceding invasive squamous cell carcinoma and basaloid carcinoma, atypical adenomatous hyperplasia, a preneoplastic condition of bronchioloalveolar carcinoma, and diffuse idiopathic pulmonary neuroendocrine cell hyperplasia, a proposed precursor for carcinoid tumours. Although the gradual accumulation of molecular alterations has been widely investigated in bronchial carcinogenesis, with the aim of determining new biomarkers for early lung cancer detection in high-risk patients and targeted chemoprevention, lung adenocarcinoma pathogenesis has been only recently highlighted, with the recent discovery of epidermal growth factor receptor mutation pathway in non-smokers. This review focuses on the current status of molecular pathology in lung cancer and pulmonary preneoplastic conditions. PMID- 19187180 TI - Advances in the understanding and classification of pulmonary hypertension. AB - Pulmonary hypertension has a complex pathobiology and the term pulmonary arterial hypertension (PAH) includes a variety of pulmonary hypertensive diseases that have different aetiologies, but which may have a similar clinical presentation and, in some cases, the same response to medical treatment. More recent classifications have been based on aetiology, functional data and clinical presentation, but morphometric studies have shown differences in the distribution and prevalence of vascular changes amongst cases of PAH of different aetiologies. Recent advances in the understanding of some aspects of the genetic and molecular mechanisms, together with detailed descriptions of the histopathological findings in PAH cases of different aetiologies, should contribute to a better understanding of the relationships of the various subsets of the disease, the prognostic implications and possible treatment options. PMID- 19187181 TI - The pathologist's approach to small airways disease. AB - The small airways constitute a significant compartment of the lung, and diseases therein are not uncommon, occurring in a wide variety of diseases, either as a primary bronchiolar disorder or as a component of local or systemic disease affecting multiple anatomical compartments. Unlike the idiopathic interstitial pneumonias, there is as yet no consensus classification system for bronchiolar disorders, and a multidisciplinary approach to their classification is discussed. Whereas some conditions show relatively characteristic histological features, others show non-specific or subtle changes that rely on a combination of clinical, radiological and pathological information for accurate final diagnosis. Furthermore, certain diseases of the small airways have been better characterized recently, and several provisional new entities have also been described, some of these being reviewed here. PMID- 19187182 TI - An approach to industrial post mortems. AB - Industrial-related deaths represent a specialized aspect of autopsy practice. The purpose of this review is to assist the pathologist in the handling of such deaths. The diseases associated with the three most significant mineral dusts (asbestos, coal and silica) are described, together with a selection of less well known mineral dust diseases. This review addresses the complex issues of ascribing disease to industrial exposures and the role of mineral analysis. The authors discuss the common medical legal issues that are encountered at post mortem and at inquest deposition. PMID- 19187183 TI - Impact of pharmacy benefit design on prescription drug utilization: a fixed effects analysis of plan sponsor data. AB - OBJECTIVE: To study the impact of various elements of pharmacy benefit design on both the absolute and relative utilization of generics, brands, retail pharmacy, and mail service. DATA SOURCE: Panel data on 1,074 plan sponsors covering 21.6 million individuals over 12 calendar quarters (2005-2007). STUDY DESIGN: A retrospective analysis of pharmacy claims. STATISTICAL METHODS: To control for potential endogeneity, linear fixed effects models were estimated for each of six dependent variables: the generic utilization rate, the brand utilization rate, the generic dispensing rate (GDR), the retail pharmacy utilization rate, the mail service utilization rate, and the mail distribution rate. PRINCIPAL FINDINGS: Most member cost-share variables were nonlinearly associated with changes in prescription drug utilization. Marginal effects were generally greater in magnitude for brand out-of-pocket costs than for generic out-of-pocket costs. Time dummies, as well as other pharmacy benefit design elements, also yielded significant results. CONCLUSIONS: Prior estimates of the effect of member cost sharing on prescription drug utilization may be biased if complex benefit designs, mail service fulfillment, and unmeasured factors such as pharmaceutical pipelines are not accounted for. Commonly cited relative utilization metrics, such as GDR, may be misleading if not examined alongside absolute prescription drug utilization. PMID- 19187184 TI - A systematic review of health care efficiency measures. AB - OBJECTIVE: To review and characterize existing health care efficiency measures in order to facilitate a common understanding about the adequacy of these methods. DATA SOURCES: Review of the MedLine and EconLit databases for articles published from 1990 to 2008, as well as search of the "gray" literature for additional measures developed by private organizations. STUDY DESIGN: We performed a systematic review for existing efficiency measures. We classified the efficiency measures by perspective, outputs, inputs, methods used, and reporting of scientific soundness. PRINCIPAL FINDINGS: We identified 265 measures in the peer reviewed literature and eight measures in the gray literature, with little overlap between the two sets of measures. Almost all of the measures did not explicitly consider the quality of care. Thus, if quality varies substantially across groups, which is likely in some cases, the measures reflect only the costs of care, not efficiency. Evidence on the measures' scientific soundness was mostly lacking: evidence on reliability or validity was reported for six measures (2.3 percent) and sensitivity analyses were reported for 67 measures (25.3 percent). CONCLUSIONS: Efficiency measures have been subjected to few rigorous evaluations of reliability and validity, and methods of accounting for quality of care in efficiency measurement are not well developed at this time. Use of these measures without greater understanding of these issues is likely to engender resistance from providers and could lead to unintended consequences. PMID- 19187185 TI - An examination of the Medicaid undercount in the current population survey: preliminary results from record linking. AB - OBJECTIVE: To assess reasons why survey estimates of Medicaid enrollment are 43 percent lower than raw Medicaid program enrollment counts (i.e., "Medicaid undercount"). DATA SOURCES: Linked 2000-2002 Medicaid Statistical Information System (MSIS) and the 2001-2002 Current Population Survey (CPS). DATA COLLECTION METHODS: Centers for Medicare and Medicaid Services provided the Census Bureau with its MSIS file. The Census Bureau linked the MSIS to the CPS data within its secure data analysis facilities. STUDY DESIGN: We analyzed how often Medicaid enrollees incorrectly answer the CPS health insurance item and imperfect concept alignment (e.g., inclusion in the MSIS of people who are not included in the CPS sample frame and people who were enrolled in Medicaid in more than one state during the year). PRINCIPAL FINDINGS: The extent to which the Medicaid enrollee data were adjusted for imperfect concept alignment reduces the raw Medicaid undercount considerably (by 12 percentage points). However, survey response errors play an even larger role with 43 percent of Medicaid enrollees answering the CPS as though they were not enrolled and 17 percent reported being uninsured. CONCLUSIONS: The CPS is widely used for health policy analysis but is a poor measure of Medicaid enrollment at any time during the year because many people who are enrolled in Medicaid fail to report it and may be incorrectly coded as being uninsured. This discrepancy should be considered when using the CPS for policy research. PMID- 19187186 TI - Use of novel selenomethionine-resistant yeast to produce selenomethionyl protein suitable for structural analysis. AB - Yeast is widely used to determine the tertiary structure of eukaryotic proteins, because of its ability to undergo post-translational modifications such as glycosylation. A mutant lacking S-adenosylmethionine synthesis has been reported as a suitable host for producing selenomethionine derivatives, which can help solve phase problems in protein crystallography. However, the mutant required external addition of S-adenosylmethionine for cell proliferation. Here, a selenomethionine-resistant Pichia pastoris mutant that showed S adenosylmethionine autotrophy was isolated. Human lysozyme expressed by the mutant under the control of constitutive promoter contained selenomethionine at 65% occupancy, sufficient for use as a selenomethionine derivative for single wavelength anomalous dispersion phasing. PMID- 19187187 TI - A novel informative dinucleotide microsatellite marker located on human factor VIII intron 25. PMID- 19187188 TI - Forum on: the role of recombinant factor VIII in children with severe haemophilia A. AB - The development of recombinant FVIII (rFVIII) products, fuelled by the need for improved safety of treatment arising from the dramatic widespread blood-borne virus transmission in the 1970-1980s revolutionized the care of children with haemophilia A over the last two decades. The larger availability of perceived safer replacement therapy associated with the introduction of rFVIII products reassured the haemophilia community and there was a strong push in some Western countries to treat haemophilic children only with rFVIII. Moreover, this significantly contributed in the 1990s to the diffusion outside Northern Europe of prophylactic regimens implemented at an early age to prevent bleeding and the resultant joint damage (i.e. primary prophylaxis), together with the possibility of home treatment. These changes led to a substantial improvement of the quality of life of haemophilic children and of their families. The general agreement that primary prophylaxis represents the first-choice treatment for haemophilic children has been recently supported by two randomized controlled trials carried out with rFVIII products, providing evidence on the efficacy of early prophylaxis over on-demand treatment in preserving joint health in haemophilic children. However, the intensity and optimal modalities of implementation of prophylaxis in children, in particular with respect to the issue of the venous access, are still debated. A number of studies also supports the role of secondary prophylaxis in children, frequently used in countries in which primary prophylaxis was introduced more recently. With viral safety now less than an issue and with the more widespread use of prophylaxis able to prevent arthropathy, the most challenging complication of replacement therapy for children with haemophilia remains the risk of inhibitor development. Despite conflicting data, there is no evidence that the type of FVIII concentrate significantly influences the complex multifactorial process leading to anti-FVIII alloantibodies, whereas other treatment-related factors are likely to increase (early intensive treatments due to surgery or severe bleeds) or reduce (prophylaxis) the risk. Although the optimal regimen is still uncertain, eradication of anti-FVIII antibodies by immune tolerance induction (ITI), usually with the same product administered at inhibitor detection, should be the first-choice treatment for all patients with recent onset inhibitors. This issue applies particularly to children, as most patients undergo ITI at an early age, when inhibitors usually appear. The availability of a stable and long-lasting venous access represents a leading problem also in this setting. These and other topics concerning rFVIII treatment of haemophilic children were discussed in a meeting held in Rome on 27 February 2008 and are summarized in this report. PMID- 19187189 TI - HCV treatment with pegylated interferon and ribavirin in patients with haemophilia and HIV/HCV co-infection. AB - Individuals with haemophilia are frequently infected with both human immunodeficiency virus (HIV) and hepatitis C virus (HCV); however, limited evidence is currently available regarding the efficacy of HCV treatment with pegylated interferon and ribavirin in this patient population. The aim of this study was to review HCV treatment outcomes in a cohort of patients with haemophilia and HIV/HCV co-infection. A retrospective, single centre review of 13 consecutive patients treated with pegylated interferon and ribavirin was performed. All patients were male with haemophilia A and a median age of 43 (range 27-62) at initiation of HCV therapy. Nine of 13 (69%) patients had genotype (gt1) 1 HCV (3 x gt3, 1 x gt4). Twelve of 13 (92%) were receiving ART, with a mean CD4+ count of 428 cells microL(-1) (range 175-928 cells microL(-1)) at initiation of HCV therapy. Six of 11 (55%) patients achieved EVR (3 x gt1, 2 x gt3, 1 x gt4) at 12 weeks, 4/13 (31%) had EOTR (2 x gt1, 2 x gt3) and 1/13 (8%) achieved sustained virological response (1 x gt1). Seven of 11 (64%) patients normalized ALT during therapy wherein mean ALT fell from 101 to 76 U L(-1). Only 1/13 (8%) patients discontinued therapy prematurely due to side effects. CD4+ cell counts and HIV viral load remained stable during HCV treatment, with a mean 437 cells microL(-1) and <50 copies mL(-1) at 48 weeks respectively. Patients in our cohort with haemophilia and HCV/HIV co-infection responded poorly to HCV treatment. Alternative HCV treatment strategies need to be considered in patients with haemophilia and HIV/HCV co-infection. PMID- 19187190 TI - Depression, anxiety and suicidal behaviour in children and adolescents with Haemophilia. AB - It was aimed to study the prevalence of major depressive disorder, separation anxiety disorder and suicidal behaviour in the children and adolescents with haemophilia. A total of 83 children and adolescents with haemophilia A or B aged 5-19 years old were randomly selected using systematic random sampling method. They were interviewed using the Farsi (Persian) version of Kiddie Schedule for Affective Disorders and Schizophrenia by face-to-face interview with the subjects and their primary caregivers, according to the DSM-IV diagnostic criteria. Major depressive disorder, separation anxiety disorder and suicidal behaviour were surveyed. The rate of major depressive disorder was 6.0%. Approximately 2.4% of the patients were suffering from at least five symptoms of major depression, and 4.8% were suffering from more than five symptoms of major depressive disorder. One-third of the children and adolescents were suffering from irritability. Approximately 36% of the subjects wished to die at least once during the last 6 months. Also, 6% of the subjects had thought about suicidal attempt. Approximately 4.8% of them met the criteria of separation anxiety disorder. The incidences of major depressive disorder and separation anxiety disorder were comparable to that prevalent in the thalassaemic patients. The proportion of patients with death wish was very high. The proportion of patients with suicidal thought and attempt was much lower than that in the children and adolescents with thalassaemia. Therefore, psychiatric consultation needs to be included in the management of these children. PMID- 19187191 TI - A systematic review of the cost-effectiveness of rFVIIa and APCC in the treatment of minor/moderate bleeding episodes for haemophilia patients with inhibitors. AB - The clinical, humanistic and economic consequences associated with haemophilia and inhibitors are considerable. Primary treatment for mild-to-moderate bleeding disorders in such patients is recombinant factor VIIa (rFVIIa) or activated prothrombin complex concentrate (APCC). The aims of this study were to identify, review and evaluate the quality of the published literature on the relative cost effectiveness of rFVIIa and APCC in treating haemophilia patients with inhibitors. The review concentrates on model type, design and assumptions, and results. The results of this study suggest that rFVIIa may be the cost-effective alternative to treatment with APCC. In seven out of the nine studies, rFVIIa had the lower average treatment cost. The difference in average treatment cost to resolve a bleed, between rFVIIa and APCC in these seven studies, ranged from $3000 to $17 000. The adapted modelling framework is similar in all the economic models reviewed, suggesting clinical acceptability of the approach used. The estimates of efficacy varied between the models, especially for APCC. The efficacy for APCC derived from retrospective studies was lower than reported in the literature. Sensitivity analysis was undertaken in the majority of the economic analyses and the results were found to be robust to realistic parameter variations. Only one of the studies was a cost-utility study, showing the lack of measuring health status within this area. This systematic review showed that models based on different sources of data produced fairly similar robust results despite differences in the estimates of efficacy, average dosage required, and unit costs. However, ideally there should be a systematic approach to identifying the relevant data. PMID- 19187192 TI - Contribution of genetical analysis for diagnosis of von Willebrand's disease type 2B. PMID- 19187193 TI - Prevalence and risk factors associated with decreased bone mineral density in patients with haemophilia. AB - Osteoporosis in adult males is an under-recognized problem. Patients with haemophilia have several predisposing factors for developing decreased bone mineral density (BMD) including prolonged periods of immobility, reduced weight bearing and co-morbidities associated with bone loss. To establish prevalence and risk factors associated with decreased BMD in patients with haemophilia. Adults with moderate or severe haemophilia A or B underwent dual-energy X-ray absorptiometry (DXA). BMD was correlated to laboratory values, joint mobility measurements and physical activity questionnaires. Thirty patients completed evaluations. The median age was 41.5 years (range 18-61). Median lowest T-score by DXA was -1.7 (range: -5.8 to +0.6), with the femoral neck being the site of the lowest T-scores. Based on World Health Organization criteria, 70% of patients had decreased BMD. Twenty-seven per cent of the participants (n = 8) had osteoporosis and 43% (n = 13) had osteopenia. Variables associated with increased bone loss included lower serum 25-hydroxyvitamin D levels (P = 0.03), lower body mass index (P = 0.047), lower activity scores (P = 0.02), decreased joint range of motion (P = 0.046), HIV (P = 0.03), HCV (P = 0.02), history of inhibitor (P = 0.01) and age (P = 0.03). Adults with haemophilia are at increased risk for developing osteoporosis. A history of HCV and HIV infections, decreased joint range-of-motion, decreased activity levels, history of an inhibitor and low body weight predict bone loss and suggest a population to target for screening. A high prevalence of vitamin D insufficiency was observed. Future studies should investigate interventions, including vitamin D supplementation, to prevent bone loss and fractures for this at-risk population. PMID- 19187194 TI - Consensus protocol for the use of recombinant activated factor VII [eptacog alfa (activated); NovoSeven] in elective orthopaedic surgery in haemophilic patients with inhibitors. AB - Patients with haemophilia complicated by inhibitors have a significant burden of joint disease, which is associated with a negative impact on their quality of life. Successful elective orthopaedic surgery can result in decreased bleed frequency into a new joint, less time spent in hospital, increased mobility and improved well being. This paper describes a new protocol for use of recombinant activated factor VII (rFVIIa) in elective orthopaedic surgery, based on a review of published data as well as the personal experience of a group of expert physicians. The protocol offers guidance on the planning of the surgery and preoperative testing as well as the bolus schedule for rFVIIa and advice on the concomitant use of antifibrinolytic agents and fibrin sealants. A total of 10 operations involving 13 procedures in eight patients in five comprehensive care centres have been undertaken until now using the protocol, which employs an initial bolus dose of rFVIIa in the range of 120-180 microg kg(-1) to cover surgery. The clinical experience reported here encompasses all cases of elective orthopaedic surgery using rFVIIa as initial treatment carried out in the UK and Republic of Ireland over the last 2 years. In all cases, there was good control of haemostasis during surgery and the final outcome was rated as 'excellent' or 'extremely satisfactory' by the reporting clinicians. Although the initial cost of product to cover surgery such as arthroplasty is high, it needs to be borne in mind that this may be offset in subsequent years by savings resulting from avoidance of bleeding episodes in the affected joint. PMID- 19187195 TI - Cumulative analysis of inhibitor formation in patients with haemophilia A treated with sucrose-formulated recombinant factor VIII. PMID- 19187196 TI - Von Willebrand disease and recurrent hematoperitoneum due to the rupture of haemorrhagic ovarian cysts. PMID- 19187197 TI - Ankle arthropathy in the haemophilic patient: a description of a novel ankle arthrodesis technique. AB - The ankle joint is well known to show early involvement in severe haemophilia. We describe a novel operative technique developed by the senior author. This combines a medial approach to the ankle, medial malleolar osteotomy, bone graft and compression with staples. All patients had excellent pain relief and improvement in function with 100% achieving bony union. There was a significant improvement in Mazur ankle scores following ankle fusion (P < 0.01). This surgical technique gives good results which are reproducible in this patient population. PMID- 19187198 TI - Molecular detection of Borrelia bissettii DNA in serum samples from patients in the Czech Republic with suspected borreliosis. AB - Until recently, three spirochete genospecies were considered to be the causative agents of Lyme borreliosis (LB) in Europe: Borrelia burgdorferi sensu stricto, Borrelia afzelii and Borrelia garinii. However, the DNA of Borrelia valaisiana, Borrelia lusitaniae, Borrelia spielmanii and Borrelia bissettii has already been detected in samples of human origin, or the spirochetes were isolated from the patients with symptoms of LB. Molecular analysis of 12 selected serum samples collected in the regional hospital confirmed the presence of B. bissettii DNA in cases of single and multiple infection in patients with symptomatic borreliosis or chronic borrelial infection. The presence of B. bissettii as a single strain in patients provides strong support of the fact that B. bissettii might be a causative agent of the disease. After the first isolation of B. bissettii from the samples of human origin in Slovenia, following the detection of this species in cardiac valve tissue of the patient with endocarditis and aortic valve stenosis in the Czech Republic, here we present additional molecular data supporting the involvement of B. bissettii in LB in Europe. PMID- 19187199 TI - Identification of a mosaic transposable element of Paracoccus marcusii composed of insertion sequence ISPmar4 (ISAs1 family) and an IS1247a-driven transposable module (TMo). AB - A sacB-based trap plasmid was used to clone a mosaic transposable element (TE) in the carotenoid producer Paracoccus marcusii OS22. This element is composed of an insertion sequence (IS) ISPmar4 (ISAs1 family), which contains an inserted functional transposable module (TMo) generated by a copy of IS1247a (of the IS1380 family). Besides IS1247a sequences, the TMo also contains the 3'-end region of a putative alpha/beta hydrolase gene, whose expression might be activated from the P(2) promoter of IS1247a. The identification of this novel TE provides evidence that transposition of TMos may change an IS into a more complex element resembling noncomposite transposons. PMID- 19187201 TI - PG27 is a novel membrane protein essential for a Porphyromonas gingivalis protease secretion system. AB - Porphyromonas gingivalis secretes endopeptidase gingipains, which are important virulence factors of this bacterium. Gingipains are transported across the inner membrane via the Sec system, followed by transport across the outer membrane via an unidentified pathway. The latter transport step is suggested to be mediated via a novel protein secretion pathway. In the present study, we report a novel candidate as an essential factor for the latter transport step. The PG0027 gene of P. gingivalis W83 encodes novel protein PG27. In a PG0027 deletion mutant (83K10), the activities of Arg-gingipain and Lys-gingipain were severely reduced, while the activities of secreted exopeptidases DPPIV, DPP-7, and PTP-A were unaffected. Protein localization was investigated by cell-surface biotinylation, subcellular fractionation, and immunoblot analysis. In the wild-type W83, Arg gingipains in membrane fraction were detected as cell surface proteins. In contrast, in 83K10, Arg-gingipains were trapped in the periplasm and hardly secreted into an extracellular milieu. PG27 was suggested to be exposed to the cell surface by a cell surface biotinylation experiment; however, PG27 was detected in both inner and outer membrane fractions by subcellular fractionation experiments. Taken together, we suggest that PG27 is a unique membrane protein essential for a novel secretion pathway. PMID- 19187200 TI - Productive Chlamydia trachomatis lymphogranuloma venereum 434 infection in cells with augmented or inactivated autophagic activities. AB - Autophagy, a eukaryotic cellular activity leading to the degradation of cellular components, serves as a defense mechanism against facultative intracellular bacteria as well as a growth niche for the obligate intracellular bacterium Coxiella burnetii. We here demonstrate that the obligate intracellular bacterial pathogen Chlamydia trachomatis lymphogranuloma venereum strongly induced autophagy in the middle of the chlamydial developmental cycle (24 h after infection), a time point with maximal level of chlamydial replication, but not during the early stages with low overall chlamydial metabolism (before 8 h). No autophagy induction was evident in cells exposed to heat- and UV-inactivated elementary bodies (EBs, the infectious form of Chlamydia) or to inocula from which EBs had been removed before inoculation. Blocking chlamydial development with chloramphenicol also prevented autophagy induction in cells infected with infectious EBs. It appears that autophagy is activated primarily in response to the metabolic stress consequent to chlamydial replication. However, autophagy defective ATG5(-/-) cells supported chlamydial development as efficiently as autophagy-proficient ATG5(+/+) cells. PMID- 19187202 TI - Enzyme-substrate interaction and characterization of a 2,3-dihydroxybiphenyl 1,2 dioxygenase from Dyella ginsengisoli LA-4. AB - A bphC gene (915 bp) encoding 2,3-dihydroxybiphenyl 1,2-dioxygenase (BphC) was amplified by PCR from Dyella ginsengisoli LA-4, which was heterologously expressed in Escherichia coli. The purified His-Tag BphC was able to catalyze the meta-cleavage reaction of the dihydroxylated aromatic rings. According to the specificity constant (K(cat)/K(m)) of BphC_LA-4, the specificity of BphC_LA-4 was determined in the following order: 2,3-dihydroxybiphenyl>3 methylcatechol>catechol>4-chlorocatechol>4-methylcatechol. The experimental data were consistent with the prediction of enzyme-substrate complexes. The highest specific activity of BphC_LA-4 was 118.3 U mg(-1) for 2,3-dihydroxybiphenyl. PMID- 19187203 TI - Characterization of a Rab11 homologue, EoRab11a, in Euplotes octocarinatus. AB - Rab GTPases are crucial in the regulation of intracellular vesicular trafficking. A novel Rab GTPase gene, EoRab11a (GenBank accession no. EF061065), was isolated and identified from Euplotes octocarinatus cells in this study. It contains an ORF of 696-bp nucleotides, encoding 231 amino acids with a calculated molecular weight of 26.8 kDa. Alignment of EoRab11a with other Rab11 proteins from other eukaryotes demonstrated that these proteins shared 53-61% identity at the amino acid level. The recombinant EoRab11a was expressed in Escherichia coli and purified by immobilized metal chelate affinity chromatography and iron chromatography. The GTPase activity of EoRab11a was 0.0024 min(-1) detected by HPLC at 30 degrees C. Three mutations were generated at amino acids Ser21 and Gly22 positions in the G1 domain of EoRab11a. All three mutants, S21P, S21G and G22R, increased the GTPase activity in vitro. Immunofluorescence microscopy results indicated that EoRab11a was localized on the phagosomal membrane during phagocytosis of E. octocarinatus. These data show that EoRab11a possesses GTP hydrolysis activity and may participate in vesicle transport events during phagocytosis of E. octocarinatus. PMID- 19187204 TI - Consequences of relative cellular positioning on quorum sensing and bacterial cell-to-cell communication. AB - Cell-to-cell bacterial communication via diffusible signals is addressed and the conceptual framework in which quorum sensing is usually described is evaluated. By applying equations ruling the physical diffusion of the autoinducer molecules, one can calculate the gradient profiles that would occur either around a single cell or at the center of volumes of increasing size and increasing cell densities. Water-based matrices at 25 degrees C and viscous biofilms at colder temperatures are compared. Some basic consequences relevant for the field of microbial signalling arise. As regards induction, gradient-mixing dynamics between as little as two cells lying at a short distance appears to be sufficient for the buildup of a concentration reaching the known thresholds for quorum sensing. A straight line in which the highest concentrations occur is also created as a consequence of the gradient overlap geometry, providing an additional signal information potentially useful for chemotactic responses. In terms of whole population signalling, it is shown how the concentration perceived by a cell in the center is critically dependent not only on the cell density but also on the size of the biofilm itself. Tables and formulas for the practical prediction of N-acyl homoserine lactones concentrations at desired distances in different cell density biofilms are provided. PMID- 19187205 TI - Characterization of temperature-sensitive and lipopolysaccharide overproducing transposon mutants of Pseudomonas putida CA-3 affected in PHA accumulation. AB - A library of 20 000 transposon (Tn5) mutants of the gram-negative bacterium Pseudomonas putida CA-3 was generated and screened for adverse affects in polyhydroxyalkanoates (PHA) accumulation. Two mutants of interest were characterized phenotypically. CA-3-126, a mutant disrupted in a stress-related protein Clp protease subunit ClpA, demonstrated greater decreases in PHA accumulation compared with the wild type at reduced and elevated temperatures under PHA-accumulating growth conditions. CA-3-M, which is affected in the aminotransferase class I enzyme, accumulated reduced levels of PHA relative to the wild type and had lower growth yields on all carbon sources tested. Mutant CA 3-M produced up to 10-fold higher levels of lipopolysaccharide relative to the wild type and exhibited 1.2-fold lower aminotransferase activity with phenylalanine as a substrate compared with the wild-type strain. The composition of the lipopolysaccharide produced by the mutant differed from that produced by the wild-type strain. Growth and PHA accumulation by CA-3-M was the same as the wild type when the nitrogen concentration in the medium was increased to 265 mg N L(-1). PMID- 19187206 TI - The cysteine 354 and 277 residues of Salmonella enterica serovar Typhi EnvZ are determinants of autophosphorylation and OmpR phosphorylation. AB - An initial biochemical characterization of the Salmonella enterica serovar Typhi (S. Typhi) EnvZ sensor protein and several mutant derivatives was performed. Autophosphorylation levels were higher for Escherichia coli EnvZ, intermediate for S. enterica serovar Typhimurium EnvZ and very low for S. Typhi EnvZ, in spite of their high amino acid sequence identity. Consequently, OmpR phosphorylation was related to EnvZ autophosphorylation. Among the mutant derivatives, a C354G mutation in S. Typhi EnvZ resulted in a substantial increase in autophosphorylation, while mutation of its other cysteine residue at position 277 to L or S decreased the EnvZ autophosphorylation level. Upon heterodimerization, the S. Typhi C354G mutant complemented the wild type in vitro, increasing the EnvZ-P yield of both monomers, in accordance with the model where EnvZ autophosphorylation occurs in trans, indicating that dimer formation is a dynamic process. Hence, the C354 and the C277 residues are fundamental in determining the particular intrinsic biochemical characteristics of EnvZ. PMID- 19187207 TI - Constitutive mineralization of low concentrations of the herbicide linuron by a Variovorax sp. strain. AB - The mineralization of the herbicide linuron at concentrations of microg and mg L( 1) was studied in liquid batch experiments with Variovorax sp. strain SRS16. The strain was highly efficient at mineralizing a range of linuron concentrations (0.002-10 mg L(-1)) with 20-60% of the added (14)C-ring-labeled linuron metabolized to (14)CO(2) within hours to days depending on the initial linuron concentration and incubation period. At mg L(-1) linuron concentrations the mineralization activity by SRS16 was inducible and a shift to constitutive mineralization activity was apparent with a reduction in the linuron concentration to microg L(-1) levels. This study revealed that strain SRS16 is a promising candidate for bioaugmentation of water or soil resources contaminated with low linuron concentrations. PMID- 19187208 TI - Manganese in biogenic magnetite crystals from magnetotactic bacteria. AB - Magnetotactic bacteria produce either magnetite (Fe(3)O(4)) or greigite (Fe(3)S(4)) crystals in cytoplasmic organelles called magnetosomes. Whereas greigite magnetosomes can contain up to 10 atom% copper, magnetite produced by magnetotactic bacteria was considered chemically pure for a long time and this characteristic was used to distinguish between biogenic and abiogenic crystals. Recently, it was shown that magnetosomes containing cobalt could be produced by three strains of Magnetospirillum. Here we show that magnetite crystals produced by uncultured magnetotactic bacteria can incorporate manganese up to 2.8 atom% of the total metal content (Fe+Mn) when manganese chloride is added to microcosms. Thus, chemical purity can no longer be taken as a strict prerequisite to consider magnetite crystals to be of biogenic origin. PMID- 19187209 TI - Novel marine carbazole-degrading bacteria. AB - Eleven carbazole (CAR)-degrading bacterial strains were isolated from seawater collected off the coast of Japan using two different media. Seven isolates were shown to be most closely related to the genera Erythrobacter, Hyphomonas, Sphingosinicella, Caulobacter, and Lysobacter. Meanwhile, strains OC3, OC6S, OC9, and OC11S showed low similarity to known bacteria, the closest relative being Kordiimonas gwangyangensis GW14-5 (90% similarity). Southern hybridization analysis revealed that only five isolates carried car genes similar to those reported in Pseudomonas resinovorans CA10 (car(CA10)) or Sphingomonas sp. strain KA1 (car(KA1)). The isolates were subjected to GC-MS and the results indicated that these strains degrade CAR to anthranilic acid. PMID- 19187210 TI - A novel type-III staphylococcal cassette chromosome mec (SCCmec) variant among Indian isolates of methicillin-resistant Staphylococcus aureus. AB - We identified a novel type-III staphylococcal cassette chromosome mec (SCCmec) element carried by eight methicillin-resistant Staphylococcus aureus (MRSA) strains from different wards and patients in an Indian hospital. Although the pulsed-field gel electrophoresis pattern and spa types of eight strains were identical and clonally related to other nosocomial Indian isolates that belonged to sequence type (ST) 239 and spa type t037, the minimum inhibitory concentration (MIC) of these eight variants was noticeably low compared with the typical type III isolates from the same hospital, and we were unable to identify ccrC and hsdR by multiplex PCR, although mer operon and transposases A, B, and C of Tn554 were amplified. By amplifying the entire SCCmec region by long-range PCR and determining parts of the nucleotide sequences of one isolate (V14), we found that the strain carried a novel SCCmec element containing a 422 bp sequence, which is highly homologous to that identified in strain CCR1-9583, mer operon and plasmid pT181 integrated in tandem via IS431 in the J3 region. It also carried a cassette chromosome, previously reported to be an SCC-like element, downstream of type-III SCCmec. Because PCR amplification of representative genes showed that these eight strains carried the same genetic elements, they belong to a novel MRSA clone that differs from most nosocomial clones carrying type-III SCCmec and SCCmercury, despite belonging to the ST239 genotype. PMID- 19187212 TI - Characterization of Salicola sp. IC10, a lipase- and protease-producing extreme halophile. AB - In order to explore the diversity of extreme halophiles able to produce different hydrolytic enzymes (amylase, protease, lipase and DNAse) in hypersaline habitats of South Spain, a screening program was performed. A total of 43 extreme halophiles showing hydrolytic activities have been isolated and characterized. The isolated strains were able to grow optimally in media with 15-20% (w/v) total salts and in most cases, growth was detected up to 30% (w/v) total salts. Most hydrolase producers were assigned to the family Halobacteriaceae, belonging to the genera Halorubrum (22 strains), Haloarcula (nine strains) and Halobacterium (nine strains), and three isolates were characterized as extremely halophilic bacteria (genera Salicola, Salinibacter and Pseudomonas). An extremely halophilic isolate, strain IC10, showing lipase and protease activities and identified as a Salicola strain of potential biotechnological interest, was further studied. The optimum growth conditions for this strain were 15-20% (w/v) NaCl, pH 8.0, and 37 degrees C. Zymographic analysis of strain IC10 detected the lipolytic activity in the intracellular fraction, showing the highest activity against p-nitrophenyl butyrate as a substrate in a colorimetric assay, whereas the proteolytic activity was detected in the extracellular fraction. This protease degraded casein, gelatin, bovine serum albumin and egg albumin. PMID- 19187211 TI - Molecular characterization and in situ quantification of anoxic arsenite oxidizing denitrifying enrichment cultures. AB - To explore the bacteria involved in the oxidation of arsenite (As(III)) under denitrifying conditions, three enrichment cultures (ECs) and one mixed culture (MC) were characterized that originated from anaerobic environmental samples. The oxidation of As(III) (0.5 mM) was dependent on NO(3) (-) addition and N(2) formation was dependent on As(III) addition. The ratio of N(2)-N formed to As(III) fed approximated the expected stoichiometry of 2.5. A 16S rRNA gene clone library analysis revealed three predominant phylotypes. The first, related to the genus Azoarcus from the division Betaproteobacteria, was found in the three ECs. The other two predominant phylotypes were closely related to the genera Acidovorax and Diaphorobacter within the Comamonadaceae family of Betaproteobacteria, and one of these was present in all of the cultures examined. FISH confirmed that Azoarcus accounted for a large fraction of bacteria present in the ECs. The Azoarcus clones had 96% sequence homology with Azoarcus sp. strain DAO1, an isolate previously reported to oxidize As(III) with nitrate. FISH analysis also confirmed that Comamonadaceae were present in all cultures. Pure cultures of Azoarcus and Diaphorobacter were isolated and shown to be responsible for nitrate-dependent As(III) oxidation. These results, taken as a whole, suggest that bacteria within the genus Azoarcus and the family Comamonadaceae are involved in the observed anoxic oxidation of As(III). PMID- 19187213 TI - Minerals in soil select distinct bacterial communities in their microhabitats. AB - We tested the hypothesis that different minerals in soil select distinct bacterial communities in their microhabitats. Mica (M), basalt (B) and rock phosphate (RP) were incubated separately in soil planted with Trifolium subterraneum, Lolium rigidum or left unplanted. After 70 days, the mineral and soil fractions were separated by sieving. Automated ribosomal intergenic spacer analysis was used to determine whether the bacterial community structure was affected by the mineral, fraction and plant treatments. Principal coordinate plots showed clustering of bacterial communities from different fraction and mineral treatments, but not from different plant treatments. Permutational multivariate anova (permanova) showed that the microhabitats of M, B and RP selected bacterial communities different from each other in unplanted and L. rigidum, and in T. subterraneum, bacterial communities from M and B differed (P<0.046). permanova also showed that each mineral fraction selected bacterial communities different from the surrounding soil fraction (P<0.05). This study shows that the structure of bacterial communities in soil is influenced by the mineral substrates in their microhabitat and that minerals in soil play a greater role in bacterial ecology than simply providing an inert matrix for bacterial growth. This study suggests that mineral heterogeneity in soil contributes to the spatial variation in bacterial communities. PMID- 19187214 TI - Isolation and molecular characterization of genetically diverse antagonistic, diazotrophic red-pigmented vibrios from different mangrove rhizospheres. AB - Genetic diversity of red-pigmented vibrios from different mangrove rhizospheres (Porteresia coarctata, Avicennia marina and Rhizophora mucronata) collected from Pichavaram mangrove, India was investigated. Twenty red-pink pigmented strains were isolated, 16S rRNA gene analyses indicted that these isolates belong to the genus Vibrio and were phylogenetically closely related to the type strains of Vibrio rhizosphaerae and Vibrio ruber. The rep-PCR analysis using GTG(5) and BOX primers had similar groupings, and segregated these pigmented Vibrio isolates including two type strains into seven unique genotypic groups (REP groups A1-A7). The rhizosphere of P. coarctata harbors highly genetically diverse groups of red pigmented vibrios compared to other plants. Multilocus sequence analysis using four genetic loci (pyrH, recA, rpoA, 16S rRNA) clearly supported the hypothesis that strains MSSRF38 (REP group A5) and MSSRF39 (REP group A6) could represent new Vibrio species. Biological functions of these vibrios were also determined and it was found that all these isolates have antagonistic activity against phytopathogens, and isolates belonging to REP groups A5 and A6 were positive for nifH gene by PCR. In conclusion, this study for the first time demonstrates the occurrence of genetically diverse groups of antagonistic, diazotrophic red pigmented vibrios from different mangrove plants and suggests a new ecological role for vibrios as heterotrophic plant associated rhizobacteria. PMID- 19187215 TI - Anaerobic degradation of the aromatic hydrocarbon biphenyl by a sulfate-reducing enrichment culture. AB - The aromatic hydrocarbon biphenyl is a widely distributed environmental pollutant. Whereas the aerobic degradation of biphenyl has been extensively studied, knowledge of the anaerobic biphenyl-oxidizing bacteria and their biochemical degradation pathway is scarce. Here, we report on an enrichment culture that oxidized biphenyl completely to carbon dioxide under sulfate reducing conditions. The biphenyl-degrading culture was dominated by two distinct bacterial species distantly affiliated with the Gram-positive genus Desulfotomaculum. Moreover, the enrichment culture has the ability to grow with benzene and a mixture of anthracene and phenanthrene as the sole source of carbon, but here the microbial community composition differed substantially from the biphenyl-grown culture. Biphenyl-4-carboxylic acid was identified as an intermediate in the biphenyl-degrading culture. Moreover, 4-fluorobiphenyl was converted cometabolically with biphenyl because in addition to the biphenyl-4 carboxylic acid, a compound identified as its fluorinated analog was observed. These findings are consistent with the general pattern in the anaerobic catabolism of many aromatic hydrocarbons where carboxylic acids are found to be central metabolites. PMID- 19187216 TI - Long term repeated prescribed burning increases evenness in the basidiomycete laccase gene pool in forest soils. AB - Repeated prescribed burning alters the biologically labile fraction of nutrients and carbon of soil organic matter (SOM). Using a long-term (30 years) repeated burning experiment where burning has been carried out at a 2- or 4-year frequency, we analysed the effect of prescribed burning on gross potential C turnover rates and phenol oxidase activity in relation to shifts in SOM composition as observed using Fourier-transform infrared spectroscopy. In tandem, we assessed the genetic diversity of basidiomycete laccases. While the overall effect of burning was a decline in phenol oxidase activity, Shannon diversity and evenness of laccases was significantly higher in burned sites. Co-correspondence analysis of SOM composition and laccase operational taxonomic unit frequency data also suggested a strong correlation. While this correlation could indicate that the observed increase in laccase genetic diversity due to burning is due to increased resource diversity, a temporal replacement of the most abundant members of the assembly by an otherwise dormant pool of fungi cannot be excluded. As such, our results fit the intermediate disturbance hypothesis. Effects were stronger in plots burned in 2-year rotations, suggesting that the 4-year burn frequency may be a more sustainable practice to ensure the long-term stability of C cycling in such ecosystems. PMID- 19187217 TI - Occurrence and patterns of antibiotic resistance in vertebrates off the Northeastern United States coast. AB - The prevalence of antibiotic-resistant bacteria in the marine environment is a growing concern, but the degree to which marine mammals, seabirds and fish harbor these organisms is not well documented. This project sought to identify the occurrence and patterns of antibiotic resistance in bacteria isolated from vertebrates of coastal waters in the northeastern United States. Four hundred and seventy-two isolates of clinical interest were tested for resistance to a suite of 16 antibiotics. Fifty-eight percent were resistant to at least one antibiotic, while 43% were resistant to multiple antibiotics. A multiple antibiotic resistance index value >or=0.2 was observed in 38% of the resistant pathogens, suggesting exposure of the animals to bacteria from significantly contaminated sites. Groups of antibiotics were identified for which bacterial resistance commonly co-occurred. Antibiotic resistance was more widespread in bacteria isolated from seabirds than marine mammals, and was more widespread in stranded or bycaught marine mammals than live marine mammals. Structuring of resistance patterns based on sample type (live/stranded/bycaught) but not animal group (mammal/bird/fish) was observed. These data indicate that antibiotic resistance is widespread in marine vertebrates, and they may be important reservoirs of antibiotic-resistant bacteria in the marine environment. PMID- 19187218 TI - Pseudomonas aeruginosa quorum-sensing signal molecules interfere with dendritic cell-induced T-cell proliferation. AB - Pseudomonas aeruginosa releases a wide array of toxins and tissue-degrading enzymes. Production of these malicious virulence factors is controlled by interbacterial communication in a process known as quorum sensing. An increasing body of evidence reveals that the bacterial signal molecule N-(3-oxododecanoyl)-L homoserine lactone (OdDHL) exhibits both quorum-sensing signalling and immune modulating properties. Recently, yet another quorum-sensing signal molecule, the Pseudomonas quinolone signal (PQS), has been shown to affect cytokine release by mitogen-stimulated human T cells. In the present article we demonstrate that both OdDHL and PQS decrease the production of interleukin-12 (IL-12) by Escherichia coli lipopolysaccharide-stimulated bone marrow-derived dendritic cells (BM-DCs) without altering their IL-10 release. Moreover, BM-DCs exposed to PQS and OdDHL during antigen stimulation exhibit a decreased ability to induce T-cell proliferation in vitro. Collectively, this suggests that OdDHL and PQS change the maturation pattern of stimulated DCs away from a proinflammatory T-helper type I directing response, thereby decreasing the antibacterial activity of the adaptive immune defence. OdDHL and PQS thus seem to possess dual activities in the infection process: as inducers of virulence factors as well as immune-modulators facilitating the infective properties of this pathogen. PMID- 19187220 TI - Highlights from the sixth meeting of the European society for Chlamydia research. PMID- 19187219 TI - A TonB-dependent outer membrane protein as a Bacteroides fragilis fibronectin binding molecule. AB - The binding of Bacteroides fragilis to plasmatic fibronectin was investigated using strains isolated from healthy subjects and from patients with bacteremia. They were cultivated in a synthetic media in which variations in cysteine concentrations determined alterations in the oxidation-reduction potential (Eh). All the strains assayed were capable of adhering to plasmatic fibronectin when cultivated under oxidizing and reducing conditions. Bacteroides fragilis 1405 showed the greatest difference when the results under these conditions were compared and it was selected for further investigations. Chemical treatments suggested the involvement of a protein in the interaction between B. fragilis and plasmatic fibronectin. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis of outer membrane proteins (OMPs) revealed differences between the extracts obtained from cultures grown under the two conditions. Protein bands of c. 102, 100, 77, 73, 50 and 40 kDa were more highly expressed under oxidizing than reducing conditions. Dot blot analysis showed a stronger recognition of plasmatic fibronectin by OMPs obtained from cultures grown under higher Eh, and Western blot assays confirmed a band of c. 102 kDa as fibronectin-binding protein. This protein was sequenced and revealed to be a putative TonB-dependent OMPs. PCR analysis confirmed the presence of this gene in all the studied strains. PMID- 19187221 TI - Analysis of proteins in Chlamydia trachomatis L2 outer membrane complex, COMC. AB - The protein composition and N-terminal sequences of proteins in the outer membrane of Chlamydia trachomatis L2 were analysed following isolation of N terminal peptides using combined fractional diagonal chromatography and identification by liquid chromatography tandem MS. Acetylation of primary amino groups of in vivo generated proteolytic cleavage sites facilitated identification of such sites in known outer membrane proteins (MOMPs). Our results further support a proposed prediction of the topology of the MOMPs. Furthermore, a previously unknown MOMP, CTL0626 (Ct372), was assigned as an MOMP with a carbohydrate-selective porin (OprB) family motif, and the presence of CTL0626 was confirmed using antibodies raised against the protein. PMID- 19187222 TI - Nucling interacts with nuclear factor-kappaB, regulating its cellular distribution. AB - Nucling is an Apaf1-binding proapoptotic protein involved in apoptosome-mediated apoptosis. Luciferase assays have revealed that the activation of nuclear factor kappaB induced by tumor necrosis factor-alpha, interleukin-1beta and lipopolysaccharide is downregulated by the overexpression of Nucling in HEK293 cells. Moreover, the expression of endogenous cyclooxygenase 2, tumor necrosis factor-alpha and galectin-3, the end-point molecules in the pathway for the activation of nuclear factor-kappaB, as well as nuclear factor-kappaB (p65) itself, is upregulated in Nucling gene-deficient mouse embryonic fibroblasts, suggesting that nuclear factor-kappaB is a target of Nucling. Subsequent study has revealed that Nucling physically interacts with nuclear factor-kappaB (p65 and p50) and that the binding domain of Nucling is its amino-terminal region (amino acids 1-466) containing ankyrin repeats. Overexpression of Nucling prevents the translocation of nuclear factor-kappaB into the nucleus. In addition, the cytoplasmic retention of endogenous nuclear factor-kappaB in resting cells is not observed in Nucling gene-deficient mouse embryonic fibroblasts. These results reveal a novel function of Nucling as a suppressor of nuclear factor-kappaB, mediated by its cytoplasmic retention through physical interaction. PMID- 19187223 TI - Fully active QAE isoform confers thermal hysteresis activity on a defective SP isoform of type III antifreeze protein. AB - Type III antifreeze protein is naturally expressed as a mixture of sulfopropyl Sephadex (SP) and quaternary aminoethyl-Sephadex (QAE)-binding isoforms, whose sequence identity is approximately 55%. We studied the ice-binding properties of a SP isoform (nfeAFP6) and the differences from those of a QAE isoform (nfeAFP8); both of these isoforms have been identified from the Japanese fish Zoarces elongatus Kner. The two isoforms possessed ice-shaping ability, such as the creation of an ice bipyramid, but nfeAFP6 was unable to halt crystal growth and exhibited no thermal hysteresis activity. For example, the ice growth rate for nfeAFP6 was 1000-fold higher than that for nfeAFP8 when measured for 0.1 mm protein solution at 0.25 degrees C below the melting point. Nevertheless, nfeAFP6 exhibited full thermal hysteresis activity in the presence of only 1% nfeAFP8 (i.e. [nfeAFP8]/[nfeAFP6] = 0.01), the effectiveness of which was indistinguishable from that of nfeAFP8 alone. We also observed a burst of ice crystal growth from the tip of the ice bipyramid for both isoforms on lowering the temperature. These results suggest that the ice growth inhibitory activity of an antifreeze protein isoform lacking the active component is restored by the addition of a minute amount of the active isoform. PMID- 19187224 TI - Transcriptome profiling analysis reveals multiple modulatory effects of Ginkgo biloba extract in the liver of rats on a high-fat diet. AB - Leaf extract of Ginkgo biloba (GBE) is increasingly used as a herbal medicine for the treatment of neurodegenerative, cardiovascular and cerebrovascular diseases. Several studies have demonstrated many protective effects of GBE in neurons, the endothelium and liver. In this study, we investigated the molecular mechanisms underlying the effects of GBE in disorders induced by long-term exposure to a high-fat diet (HFD). Rats were fed an HFD with or without the GBE product GBE50 for 19 weeks. We found that GBE50 reduced the development of fatty liver induced by an HFD and inhibited the commonly observed elevation of serum cholesterol and lactate dehydrogenase levels. Transcriptome profiling analysis showed that several genes were modulated by GBE50 in liver, including those involved in lipid metabolism, carbohydrate metabolism, vascular constriction, ion transportation, neuronal systems and drug metabolism. Notably, a number of genes coding for proteins involved in cholesterol metabolism were repressed, and some were upregulated. Fatty acid biosynthesis appeared to be repressed, whereas fatty acid metabolism appeared to be enhanced. In conclusion, using transcriptome profiling analysis, we demonstrated the molecular basis for the pleiotropic effects of GBE50, particularly those involved in lipid metabolism. This study provided new clues for further pharmacological study of GBEs. PMID- 19187225 TI - E2A participates in a fine control of pre-mature B-cell apoptosis mediated by B cell receptor signaling via transcriptional regulation of survivin, IAP2 and caspase-8 genes. AB - Antigen binding to the B-cell receptor (BCR) of pre-mature B lymphocytes induces their apoptotic cell death, but binding to the BCR of mature B lymphocytes triggers activation and proliferation. Binding to pre-mature B lymphocytes is thought not only to function as a mechanism to exclude B-cell clones that possess the ability to react with self-antigen, but also to act as a defense mechanism in auto-immune diseases. Cross-linking of BCR of pre-mature B-cell lines, including the chicken DT40 cell line, with anti-immunoglobulin IgG induces apoptotic cell death. Treatment with phorbol 12-myristate 13-acetate/ionomycin, which mimics BCR stimulation, is used to study intracellular signal transduction of B lymphocytes. Here, by analyzing the E2A-deficient DT40 cell line, E2A(-/-), we show that E2A deficiency prevents certain levels of apoptotic cell death mediated by BCR signaling. In addition, E2A deficiency-linked BCR signaling controls the mimicked pre-mature B-cell apoptosis by PMA/ionomycin through elevated survivin plus inhibitor of apoptosis 2 levels, and reduced caspase-3 and caspase-8 activities, resulting in increased amounts of ICAD (inhibitor of caspase-activated DNase), compared with those in the presence of E2A, followed by reduction of DNA fragmentation. These findings will contribute to the resolution of molecular mechanisms of negative selection of B cells and also auto-immune diseases. PMID- 19187226 TI - Hypoxia induces erythroid-specific 5-aminolevulinate synthase expression in human erythroid cells through transforming growth factor-beta signaling. AB - Hypoxia induces expansion of erythroid precursor cells through erythropoietin production. However, it has also been suggested that hypoxia could enhance hemoglobin production in erythroid cells directly. To identify the molecules that are involved in hemoglobin production under hypoxia, we examined the expression profile of mRNAs in YN-1 human erythroleukemia cells under hypoxia. DNA array analysis revealed that the expression of transforming growth factor (TGF)-beta1 and mitoferrin, which is a mitochondrial iron transporter, was induced after 6 h under hypoxia in YN-1 cells, whereas the increased expression of erythroid specific 5-aminolevulinate synthase (ALAS2) and gamma-globin mRNAs was observed after 48 h. Further analysis revealed that hypoxia enhanced the accumulation of TGF-beta1 in the culture medium of cells of the YN-1-0-A line, which was a clonal variant of YN-1 and could be maintained in serum-free medium. Moreover, exogenous TGF-beta1 induced hemoglobinization and the expression of ALAS2 mRNA in YN-1-0-A cells, but not of gamma-globin and mitoferrin mRNAs. Importantly, a specific inhibitor of intracellular TGF-beta signaling markedly reduced the degree of the hypoxia-mediated increase in the expression of ALAS2 mRNA in YN-1-0-A cells. On the other hand, nonhypoxic inducer of hypoxia-inducible factor 1 increased the expression of mitoferrin mRNA but not of TGF-beta1 mRNA in YN-1 cells under normoxia, suggesting that mitoferrin mRNA expression may be regulated by hypoxia inducible factor 1. Thus, our data suggest that hypoxia induces the expression of TGF-beta1 and mitoferrin mRNAs through separate mechanisms in erythroid cells. TGF-beta1 subsequently induces ALAS2 expression, which may contribute to terminal differentiation of erythroid cells. PMID- 19187227 TI - Structure of the atrial natriuretic peptide receptor extracellular domain in the unbound and hormone-bound states by single-particle electron microscopy. AB - Atrial natriuretic peptide (ANP) plays a major role in blood pressure and volume regulation. ANP activities are mediated by a cell surface, single-span transmembrane receptor linked to its intrinsic guanylate cyclase activity. The crystal structures of the dimerized ANP receptor extracellular domain (ECD) with and without ANP have revealed a novel hormone-induced rotation mechanism occurring in the juxtamembrane region that appears to mediate signal transduction [Ogawa H, Qiu Y, Ogata CM & Misono KS (2004) J Biol Chem 279, 28625-28631]. However, the ECD crystal packing contains two major intermolecular contacts that suggest two possible dimer pairs: 'head-to-head' (hh) and 'tail-to-tail' (tt) dimers associated via the membrane-distal and membrane-proximal subdomains, respectively. The existence of these two potential dimer forms challenges the proposed signaling mechanism. In this study, we performed single-particle electron microscopy (EM) to determine the ECD dimer structures occurring in the absence of crystal contacts. EM reconstruction yielded the dimer structures with and without ANP in only the hh dimer forms. We further performed steady-state fluorescence spectroscopy of Trp residues, one of which (Trp74) occurs in the hh dimer interface and none of which occurs in the tt dimer interface. ANP binding caused a time-dependent decrease in Trp emission at 350 nm that was attributable to partially buried Trp74 in the unbound hh dimer interface becoming exposed to solvent water upon ANP binding. Thus, the results of single-particle EM and Trp fluorescence studies have provided direct evidence for hh dimer structures for unbound and ANP-bound receptor. The results also support the proposed rotation mechanism for transmembrane signaling by the ANP receptor. PMID- 19187228 TI - Novel modified version of nonphosphorylated sugar metabolism--an alternative L rhamnose pathway of Sphingomonas sp. AB - Several bacteria, including Azotobacter vinelandii, possess an alternative pathway of L-rhamnose metabolism, which is different from the known bacterial pathway. In a previous article, a gene cluster related to this pathway was identified, consisting of the genes encoding the four metabolic enzymes L rhamnose-1-dehydrogenase (LRA1), L-rhamnono-gamma-lactonase (LRA2), L-rhamnonate dehydratase (LRA3) and L-2-keto-3-deoxyrhamnonate (L-KDR) aldolase (LRA4), by which L-rhamnose is converted into pyruvate and L-lactaldehyde, through analogous reaction steps to the well-known Entner-Doudoroff (ED) pathway. In this study, bioinformatic analysis revealed that Sphingomonas sp. possesses a gene cluster consisting of LRA1-3 and two genes of unknown function, LRA5 and LRA6. LRA5 catalyzed the NAD(+)-dependent dehydrogenation of several L-2-keto-3-deoxyacid sugars, including L-KDR. Furthermore, the reaction product was converted to pyruvate and L-lactate by LRA6; this is different from the pathway of Azotobacter vinelandii. Therefore, LRA5 and LRA6 were assigned as the novel enzymes L-KDR 4 dehydrogenase and L-2,4-diketo-3-deoxyrhamnonate hydrolase, respectively. Interestingly, both enzymes were phylogenetically similar to L-rhamnose-1 dehydrogenase and D-2-keto-3-deoxyarabinonate dehydratase, respectively, and the latter was involved in the archeal nonphosphorylative d-arabinose pathway, which is partially analogous to the ED pathway. The introduction of LRA1-4 or LRA1-3, LRA5 and LAR6 compensated for the L-rhamnose-defective phenotype of an Escherichia coli mutant. Metabolic evolution and promiscuity between the alternative l-rhamnose pathway and other sugar pathways analogous to the ED pathway are discussed. PMID- 19187229 TI - Hypoxia downregulates farnesoid X receptor via a hypoxia-inducible factor independent but p38 mitogen-activated protein kinase-dependent pathway. AB - Farnesoid X receptor (FXR), a member of the nuclear receptor superfamily, has been shown to play pivotal roles in bile acid homeostasis by regulating the biosynthesis, conjugation, secretion and absorption of bile acids. Accumulating data suggest that FXR signaling is involved in the pathogenesis of liver and metabolic disorders. Here we show that FXR expression is significantly suppressed in HepG2 cells exposed to hypoxia. Concomitantly, the expression of the bile salt export pump, known as an FXR target gene product and responsible for the excretion of bile acids from the liver, is also decreased under hypoxia. Overexpression of hypoxia-inducible factor (HIF)-1alpha does not mimic the suppressive effect of hypoxia on FXR expression. Furthermore, simultaneous knockdown of HIF-1alpha, HIF-2alpha and HIF-3alpha fails to restore the FXR expression level under hypoxia, indicating that HIF is not involved in hypoxia evoked FXR downregulation. Instead, we demonstrate that p38 mitogen-activated protein kinase is an indispensable factor for FXR downregulation under hypoxia. Thus, we propose a novel liver disorder model in which two signaling molecules, p38 mitogen-activated protein kinase and FXR, may contribute to the linkage of two pathogenic conditions, i.e. ischemia, a condition accompanying hypoxia, and cholestasis, a condition with intrahepatic accumulation of cytotoxic bile acids. PMID- 19187230 TI - Amino acid discrimination by arginyl-tRNA synthetases as revealed by an examination of natural specificity variants. AB - L-canavanine occurs as a toxic non-protein amino acid in more than 1500 leguminous plants. One mechanism of its toxicity is its incorporation into proteins, replacing L-arginine and giving rise to functionally aberrant polypeptides. A comparison between the recombinant arginyl-tRNA synthetases from a canavanine producer (jack bean) and from a related non-producer (soybean) provided an opportunity to study the mechanism that has evolved to discriminate successfully between the proteinogenic amino acid and its non-protein analogue. In contrast to the enzyme from jack bean, the soybean enzyme effectively produced canavanyl-tRNA(Arg) when using RNA transcribed from the jack bean tRNA(ACG) gene. The corresponding k(cat)/K(M) values gave a discrimination factor of 485 for the jack bean enzyme. The arginyl-tRNA synthetase does not possess hydrolytic post transfer editing activity. In a heterologous system containing either native Escherichia coli tRNA(Arg) or the modification-lacking E. coli transcript RNA, efficient discrimination between L-arginine and L-canavanine by both plant enzymes (but not by the E. coli arginyl-tRNA synthetase) occurred. Thus, interaction of structural features of the tRNA with the enzyme plays a significant role in determining the accuracy of tRNA arginylation. Of the potential amino acid substrates tested, apart from L-canavanine, only L thioarginine was active in aminoacylation. As it is an equally good substrate for the arginyl-tRNA synthetase from both plants, it is concluded that the higher discriminatory power of the jack bean enzyme towards L-canavanine does not necessarily provide increased protection against analogues in general, but appears to have evolved specifically to avoid auto-toxicity. PMID- 19187231 TI - Autophagy inhibits reactive oxygen species-mediated apoptosis via activating p38 nuclear factor-kappa B survival pathways in oridonin-treated murine fibrosarcoma L929 cells. AB - Autophagy and apoptosis have been known to be interconnected positively or negatively; however, the molecular mechanisms mediating these two cellular processes are not fully understood. In the present study, we demonstrated that the exposure of L929 cells to oridonin led to intracellular reactive oxygen species generation, followed by lipid peroxidation, as well as decreases in superoxide dismutase and glutathione activities. The reactive oxygen species scavenger N-acetyl-cysteine resulted in the complete inhibition of oridonin induced apoptosis and mitochondrial membrane potential collapse. We showed that reactive oxygen species triggered apoptosis by Bax translocation, cytochrome c release and extracellular signal-regulated kinase activation. Further data confirmed that oridonin also induced L929 cell autophagy, as demonstrated by extensive autophagic vacuolization and the punctuate distribution of monodansylcadaverine staining and GFP-LC3, as well as the LC3-II/LC3-I proportion and Beclin 1 activation. Subsequently, we found that inhibition of autophagy by 3 methyladenine or small interfering RNA against LC3 and Beclin 1 promoted oridonin induced cell apoptosis. The effects of p38 and nuclear factor-kappa B in oridonin induced apoptosis and autophagy were further examined. Interruption of p38 and nuclear factor-kappa B activation by specific inhibitors or small interfering RNAs promoted apoptosis and reactive oxygen species generation, but decreased autophagy. Moreover, we showed that inhibition of autophagy reduced oridonin induced activation of p38. Additionally, nuclear factor-kappa B activation was inhibited by blocking the p38 pathway. Consequently, these findings indicate that oridonin-induced L929 cell apoptosis is regulated by reactive oxygen species mediated signaling pathways, and that oridonin-induced autophagy may block apoptosis by up-regulating p38 and nuclear factor-kappa B activation. PMID- 19187232 TI - Crystal structures of two novel sulfonylurea herbicides in complex with Arabidopsis thaliana acetohydroxyacid synthase. AB - Acetohydroxyacid synthase (AHAS; EC 2.2.1.6) is the first enzyme in the biosynthetic pathway of the branched-chain amino acids. It catalyzes the conversion of two molecules of pyruvate into 2-acetolactate or one molecule of pyruvate and one molecule of 2-ketobutyrate into 2-aceto-2-hydroxybutyrate. AHAS requires the cofactors thiamine diphosphate (ThDP), Mg(2+) and FAD for activity. The herbicides that target this enzyme are effective in protecting a broad range of crops from weed species. However, resistance in the field is now a serious problem worldwide. To address this, two new sulfonylureas, monosulfuron and monosulfuron ester, have been developed as commercial herbicides in China. These molecules differ from the traditional sulfonylureas in that the heterocyclic ring attached to the nitrogen atom of the sulfonylurea bridge is monosubstituted rather than disubstituted. The structures of these compounds in complex with the catalytic subunit of Arabidopsis thaliana AHAS have been determined to 3.0 and 2.8 A, respectively. In both complexes, these molecules are bound in the tunnel leading to the active site, such that the sole substituent of the heterocyclic ring is buried deepest and oriented towards the ThDP. Unlike the structures of Arabidopsis thaliana AHAS in complex with the classic disubstituted sulfonylureas, where ThDP is broken, this cofactor is intact and present most likely as the hydroxylethyl intermediate. PMID- 19187233 TI - Protein transport in organelles: Dual targeting of proteins to mitochondria and chloroplasts. AB - As many as fifty proteins have now been experimentally demonstrated to be targeted to both mitochondria and plastids, a phenomenon referred to as dual targeting. Although the first reported case of dual targeting of a protein was reported in 1995, there is still little understanding of the mechanism of dual targeting and any similarities or differences with respect to the targeting of location-specific proteins. This minireview summarizes dual targeting in terms of signals, passenger proteins, receptors, regulation, why proteins may need to be dual targeted and the future challenges that remain in this area. PMID- 19187234 TI - Protein transport in organelles: Protein transport into and across the thylakoid membrane. AB - The chloroplast thylakoid is the most abundant membrane system in nature, and is responsible for the critical processes of light capture, electron transport and photophosphorylation. Most of the resident proteins are imported from the cytosol and then transported into or across the thylakoid membrane. This minireview describes the multitude of pathways used for these proteins. We discuss the huge differences in the mechanisms involved in the secretory and twin-arginine translocase pathways used for the transport of proteins into the lumen, with an emphasis on the differing substrate conformations and energy requirements. We also discuss the rationale for the use of two different systems for membrane protein insertion: the signal recognition particle pathway and the so-called spontaneous pathway. The recent crystallization of a key chloroplast signal recognition particle component provides new insights into this rather unique form of signal recognition particle. PMID- 19187235 TI - Protein transport in organelles: The composition, function and regulation of the Tic complex in chloroplast protein import. AB - It is widely accepted that chloroplasts derived from an endosymbiotic event in which an early eukaryotic cell engulfed an ancient cyanobacterial prokaryote. During subsequent evolution, this new organelle lost its autonomy by transferring most of its genetic information to the host cell nucleus and therefore became dependent on protein import from the cytoplasm. The so-called 'general import pathway' makes use of two multisubunit protein translocases located in the two envelope membranes: the Toc and Tic complexes (translocon at the outer/inner envelope membrane of chloroplasts). The main function of both complexes, which are thought to work in parallel, is to provide a protein-selective channel through the envelope membrane and to exert the necessary driving force for the translocation. To achieve high efficiency of protein import, additional regulatory subunits have been developed that sense, and quickly react to, signals giving information about the status and demand of the organelle. These include calcium-mediated signals, most likely through a potential plastidic calmodulin, as well as redox sensing (e.g. via the stromal NADP(+)/NADPH pool). In this minireview, we briefly summarize the present knowledge of how the Tic complex adapted to the tasks outlined above, focusing more on the recent advances in the field, which have brought substantial progress concerning the motor function as well as the regulatory potential of this protein translocation system. PMID- 19187236 TI - Protein transport in organelles: The Toc complex way of preprotein import. AB - Most of the estimated 1000 or so chloroplast proteins are synthesized as cytosolic preproteins with N-terminal cleavable targeting sequences (transit peptide). Translocon complexes at the outer (Toc) and inner chloroplast envelope membrane (Tic) concertedly facilitate post-translational import of preproteins into the chloroplast. Three components, the Toc34 and Toc159 GTPases together with the Toc75 channel, form the core of the Toc complex. The two GTPases act as GTP-dependent receptors at the chloroplast surface and promote insertion of the preprotein across the Toc75 channel. Additional factors guide preproteins to the Toc complex or support their stable ATP-dependent binding to the chloroplast. This minireview describes the components of the Toc complex and their function during the initial steps of preprotein translocation across the chloroplast envelope. PMID- 19187237 TI - Protein transport in organelles. PMID- 19187238 TI - Solution structure of the bb' domains of human protein disulfide isomerase. AB - Protein disulfide isomerase is the most abundant and best studied of the disulfide isomerases that catalyze disulfide bond formation in the endoplasmic reticulum, yet the specifics of how it binds substrate have been elusive. Protein disulfide isomerase is composed of four thioredoxin-like domains (abb'a'). Cross linking studies with radiolabeled peptides and unfolded proteins have shown that it binds incompletely folded proteins primarily via its third domain, b'. Here, we determined the solution structure of the second and third domains of human protein disulfide isomerase (b and b', respectively) by triple-resonance NMR spectroscopy and molecular modeling. NMR titrations identified a large hydrophobic surface within the b' domain that binds unfolded ribonuclease A and the peptides mastoparan and somatostatin. Protein disulfide isomerase-catalyzed refolding of reduced ribonuclease A in vitro was inhibited by these peptides at concentrations equal to their affinity to the bb' fragment. Our findings provide a structural basis for previous kinetic and cross-linking studies which have shown that protein disulfide isomerase exhibits a saturable, substrate-binding site. PMID- 19187239 TI - Hansenula polymorpha pex11 cells are affected in peroxisome retention. AB - We have cloned and characterized the Hansenula polymorpha PEX11 gene. Our morphological data are consistent with previous observations that peroxisome proliferation can be regulated by modulating Pex11p levels. Surprisingly, pex11 cells also showed a defect in peroxisome retention in mother cells during vegetative cell reproduction. Until now, Saccharomyces cerevisiae Inp1p has been the only peroxisomal protein that has been shown to play a role in the organelle retention process. H. polymorpha inp1 cells are also affected in peroxisome retention, like pex11 cells. We show by time-lapse imaging that Inp1-green fluorescent protein localization varies during the cell cycle and that the protein is normally recruited to peroxisomes in pex11 cells. Taken together, our data show that H. polymorpha Pex11p is not only important for peroxisome proliferation but is also required for proper peroxisome segregation during cell division. PMID- 19187240 TI - Kinetic and mechanistic characterization of Mycobacterium tuberculosis glutamyl tRNA synthetase and determination of its oligomeric structure in solution. AB - Mycobacterium tuberculosis glutamyl-tRNA synthetase (Mt-GluRS), encoded by Rv2992c, was overproduced in Escherichia coli cells, and purified to homogeneity. It was found to be similar to the other well-characterized GluRS, especially the E. coli enzyme, with respect to the requirement for bound tRNA(Glu) to produce the glutamyl-AMP intermediate, and the steady-state kinetic parameters k(cat) (130 min(-1)) and K(M) for tRNA (0.7 microm) and ATP (78 microm), but to differ by a one order of magnitude higher K(M) value for L-Glu (2.7 mm). At variance with the E. coli enzyme, among the several compounds tested as inhibitors, only pyrophosphate and the glutamyl-AMP analog glutamol-AMP were effective, with K(i) values in the mum range. The observed inhibition patterns are consistent with a random binding of ATP and L-Glu to the enzyme-tRNA complex. Mt-GluRS, which is predicted by genome analysis to be of the non-discriminating type, was not toxic when overproduced in E. coli cells indicating that it does not catalyse the mischarging of E. coli tRNA(Gln) with L-Glu and that GluRS/tRNA(Gln) recognition is species specific. Mt-GluRS was significantly more sensitive than the E. coli form to tryptic and chymotryptic limited proteolysis. For both enzymes chymotrypsin-sensitive sites were found in the predicted tRNA stem contact domain next to the ATP binding site. Mt-GluRS, but not Ec-GluRS, was fully protected from proteolysis by ATP and glutamol-AMP. Small-angle X-ray scattering showed that, at variance with the E. coli enzyme that is strictly monomeric, the Mt GluRS monomer is present in solution in equilibrium with the homodimer. The monomer prevails at low protein concentrations and is stabilized by ATP but not by glutamol-AMP. Inspection of small-angle X-ray scattering-based models of Mt GluRS reveals that both the monomer and the dimer are catalytically active. By using affinity chromatography and His(6)-tagged forms of either GluRS or glutamyl tRNA reductase as the bait it was shown that the M. tuberculosis proteins can form a complex, which may control the flux of Glu-tRNA(Glu) toward protein or tetrapyrrole biosynthesis. PMID- 19187241 TI - Tamavidins--novel avidin-like biotin-binding proteins from the Tamogitake mushroom. AB - Novel biotin-binding proteins, referred to herein as tamavidin 1 and tamavidin 2, were found in a basidiomycete fungus, Pleurotus cornucopiae, known as the Tamogitake mushroom. These are the first avidin-like proteins to be discovered in organisms other than birds and bacteria. Tamavidin 1 and tamavidin 2 have amino acid sequences with 31% and 36% identity, respectively, to avidin, and 47% and 48% identity, respectively, to streptavidin. Unlike any other biotin-binding proteins, tamavidin 1 and tamavidin 2 are expressed as soluble proteins at a high level in Escherichia coli. Recombinant tamavidin 2 was purified as a tetrameric protein in a single step by 2-iminobiotin affinity chromatography, with a yield of 5 mg per 100 mL culture of E. coli. The kinetic parameters measured by a BIAcore biosensor indicated that recombinant tamavidin 2 binds biotin with high affinity, in a similar manner to binding by avidin and streptavidin. The overall crystal structure of recombinant tamavidin 2 is similar to that of avidin and streptavidin. However, recombinant tamavidin 2 is immunologically distinct from avidin and streptavidin. Tamavidin 2 and streptavidin are very similar in terms of the arrangement of the residues interacting with biotin, but different with regard to the number of hydrogen bonds to biotin carboxylate. Recombinant tamavidin 2 is more stable than avidin and streptavidin at high temperature, and nonspecific binding to DNA and human serum by recombinant tamavidin 2 is lower than that for avidin. These findings highlight tamavidin 2 as a probable powerful tool, in addition to avidin and streptavidin, in numerous applications of biotin binding proteins. PMID- 19187242 TI - Molecular characterization and gene disruption of mouse lysosomal putative serine carboxypeptidase 1. AB - The retinoid-inducible serine carboxypeptidase 1 (Scpep1; formerly RISC) is a lysosomal matrix protein that was initially identified in a screen for genes induced by retinoic acid. Recently, it has been spotlighted by several proteome analyses of the lysosomal compartment, but its cellular function and properties remain unknown to date. In this study, Scpep1 from mice was analysed with regard to its intracellular processing into a mature dimer consisting of a 35 kDa N terminal fragment and a so far unknown 18 kDa C-terminal fragment and the glycosylation status of the mature Scpep1 fragment. Although Scpep1 shares notable homology and a number of structural hallmarks with the well-described lysosomal carboxypeptidase protective protein/cathepsin A, the purified recombinant 55 kDa precursor and the homogenates of Scpep1-overexpressing cells do not show proteolytic activity or increased serine carboxypeptidase activity towards artificial serine carboxypeptidase substrates. Hence, we disrupted the Scpep1 gene in mice by a gene trap cassette, resulting in a Scpep1/beta galactosidase/neomycin phosphotransferase fusion protein. The fusion protein is devoid of the C-terminal half of Scpep1, including two amino acids of the assumed catalytic triad which is indispensable for its predicted serine carboxypeptidase activity. However, Scpep1-deficient mice were viable and fertile, and did not exhibit either lysosomal storage or reduced lysosomal SC activity under any tested condition. PMID- 19187243 TI - Interactions between environmental stress and male mating success may enhance evolutionary divergence of stress-resistant Drosophila populations. AB - Adaptation of natural and laboratory-selected populations of Drosophila to desiccation stress results in enhanced water conservation abilities, and thus increased stress resistance. In this study, we tested whether laboratory selection for desiccation resistance is also reflected in increased mating success of adapted D. melanogaster males under desiccating conditions. Adapted flies perform better under stressful conditions, and as expected males from desiccation-selected populations exhibited significantly higher relative mating success in comparison with controls after 5-6 h of desiccation. However, we show evidence for a trade-off between survival under stressful conditions and mating success in nonstressful and even mildly stressful environments (2.5-3 h of desiccation), where males from selected populations were involved in only approximately 40% of observed copulations. This suggests that mutations favored by natural selection, associated with survival when resources are limited, may only be favored by sexual selection above a minimal "threshold" stress level. At milder stress levels increased resistance comes at a cost of lower relative mating success, and thus reduced fitness. This interaction between stress and relative male mating success of adapted and nonadapted males could interrupt gene flow, thus facilitating divergence of resistant populations from the ancestral population. PMID- 19187244 TI - Costs and limits of phenotypic plasticity in island populations of the common frog Rana temporaria under divergent selection pressures. AB - Costs and limits are assumed to be the major constraints on the evolution of phenotypic plasticity. However, despite their expected importance, they have been surprisingly hard to find in natural populations. It has therefore been argued that natural selection might have removed high-cost genotypes in all populations. However, if costs of plasticity are linked to the degree of plasticity expressed, then high costs of plasticity would only be present in populations where increased plasticity is under selection. We tested this hypothesis by investigating costs and limits of adaptive phenotypic plasticity in development time in a common garden study of island populations of the common frog Rana temporaria, which have varying levels of development time and phenotypic plasticity. Costs of plasticity were only found in populations with high-plastic genotypes, whereas the populations with the most canalized genotypes instead had a cost of canalization. Moreover, individuals displaying the most extreme phenotypes also were the most plastic ones, which mean we found no limits of plasticity. This suggests that costs of plasticity increase with increased level of plasticity in the populations, and therefore costs of plasticity might be more commonly found in high-plastic populations. PMID- 19187245 TI - Experimental evidence for frequency dependent self-fertilization in the gynodioecious plant, Silene vulgaris. AB - After over a half century of empirical and theoretical research regarding the evolution and maintenance of gynodioecy in plants, unexplored factors influencing the relative fitnesses of females and hermaphrodites remain. Theoretical studies suggest that hermaphrodite self-fertilization (selfing) rate influences the maintenance of gynodioecy and we hypothesized that population sex ratio may influence hermaphrodite selfing rate. An experimental test for frequency dependent self-fertilization was conducted using replicated populations constructed with different sex ratios of the gynodioecious plant Silene vulgaris. We found that hermaphrodite selfing increased with decreased hermaphrodite frequency, whereas evidence for increased inbreeding depression was equivocal. We argue that incorporation of context dependent inbreeding into future models of the evolution of gynodioecy is likely to yield novel insights into sex ratio evolution. PMID- 19187246 TI - Easy and flexible Bayesian inference of quantitative genetic parameters. AB - There has been a tremendous advancement of Bayesian methodology in quantitative genetics and evolutionary biology. Still, there are relatively few publications that apply this methodology, probably because the availability of multipurpose and user-friendly software is somewhat limited. It is here described how only a few rows of code of the well-developed and very flexible Bayesian software WinBUGS (Lunn et al. 2000) can be used for inference of the additive polygenic variance and heritabilty in pedigrees of general design. The presented code is illustrated by application to an earlier published dataset of Scots pine. PMID- 19187247 TI - Migration-selection balance and local adaptation of mitochondrial haplotypes in rufous-collared sparrows (Zonotrichia capensis) along an elevational gradient. AB - Variable selection pressures across heterogeneous landscapes can lead to local adaptation of populations. The extent of local adaptation depends on the interplay between natural selection and gene flow, but the nature of this relationship is complex. Gene flow can constrain local adaptation by eroding differentiation driven by natural selection, or local adaptation can itself constrain gene flow through selection against maladapted immigrants. Here we test for evidence that natural selection constrains gene flow among populations of a widespread passerine bird (Zonotrichia capensis) that are distributed along an elevational gradient in the Peruvian Andes. Using multilocus sequences and microsatellites screened in 142 individuals collected along a series of replicate transects, we found that mitochondrial gene flow was significantly reduced along elevational transects relative to latitudinal control transects. Nuclear gene flow, however, was not similarly reduced. Clines in mitochondrial haplotype frequency were strongly associated with transitions in environmental variables along the elevational transects, but this association was not observed for the nuclear markers. These results suggest that natural selection constrains mitochondrial gene flow along elevational gradients and that the mitonuclear discrepancy may be due to local adaptation of mitochondrial haplotypes. PMID- 19187248 TI - Does variation in selection imposed by bears drive divergence among populations in the size and shape of sockeye salmon? AB - Few studies have determined whether formal estimates of selection explain patterns of trait divergence among populations, yet this is one approach for evaluating whether the populations are in equilibria. If adaptive divergence is complete, directional selection should be absent and stabilizing selection should prevail. We estimated natural selection, due to bear predation, acting on the body size and shape of male salmon in three breeding populations that experience differing predation regimes. Our approach was to (1) estimate selection acting within each population on each trait based on an empirical estimate of reproductive activity, (2) test for trait divergence among populations, and (3) test whether selection coefficients were correlated with trait divergence among populations. Stabilizing selection was never significant, indicating that these populations have yet to attain equilibria. Directional selection varied among populations in a manner consistent with trait divergence, indicating ongoing population differentiation. Specifically, the rank order of the creeks in terms of patterns of selection paralleled the rank order in terms of size and shape. The shortest and least deep-bodied males had the highest reproductive activity in the creek with the most intense predation and longer and deeper-bodied males were favored in the creeks with lower predation risk. PMID- 19187249 TI - Host specialization differentiates cryptic species of feather-feeding lice. AB - Parasite species with differentiated host-specific populations provide a natural opportunity to explore factors involved in parasite diversification. Columbicola macrourae is a species of ectoparasitic feather louse currently recognized from 15 species of New World pigeons and doves. Mitochondrial sequences reveal five divergent haplotype clusters within C. macrourae, suggesting cryptic species. Each cluster is relatively host specific, with only one or a few hosts. We conducted a reciprocal transfer experiment with two of these lineages to test whether host use has an adaptive component. Our results demonstrate that the fitness of each lineage is considerably higher on its native host than on the novel host suggesting that one or more selective agents favor host specialization by the different lineages. In addition, we were able to morphologically separate individual lice from the two experimental lineages using discriminant function analysis. Furthermore, differences in the size of these louse lineages match differences in the size of their respective hosts, paralleling the strong correlation between parasite and host body size across the genus Columbicola. Together, these results suggest that selection in this cryptic species complex reflects selection across the whole genus, and that this selection, in part, contributes to the maintenance of host specialization. PMID- 19187250 TI - Genetic correlations in a wild rodent: grass-eaters and fast-growers evolve high basal metabolic rates. AB - Basal metabolic rate (BMR), commonly used as a measure of the cost of living, is highly variable among species, and sources of the variation are subject to an enduring debate among comparative biologists. One of the hypotheses links the variation in BMR with diversity of food habits and life-history traits. We test this hypothesis by asking how BMR of a particular species, the bank vole Myodes (=Clethrionomys) glareolus, would change under selection for high growth rate (measured as a postweaning body mass change; MD(PW)) and the ability to cope with a low-quality herbivorous diet (measured as body mass change during a four-day test; MD(LQD)). We show that both of the traits are heritable in the narrow sense (MD(PW): h(2)= 0.30; MD(LQD): h(2)= 0.19), and are genetically correlated with mass-independent BMR (additive genetic correlation, r(A)= 0.28 for MD(PW) and 0.37 for MD(LQD)). Thus, both of the traits could change in response to a selection, and the selection would also result in a correlated evolution of the level of metabolism. The results are consistent with the hypothesis that a part of the interspecific variation in BMR evolved in response to selection for life history and ecological traits such as food habits. PMID- 19187251 TI - Molecular clocks provide new insights into the evolutionary history of Galeichthyine sea catfishes. AB - Intercontinental distributions in the southern hemisphere can either be the result of Gondwanan vicariance or more recent transoceanic dispersal. Transoceanic dispersal has come into vogue for explaining many intercontinental distributions; however, it has been used mainly for organisms that can float or raft between the continents. Despite their name, the Sea Catfishes (Ariidae) have limited dispersal ability, and there are no examples of nearshore ariid genera with a transoceanic distribution except for Galeichthys where three species occur in southern Africa and one in the Peruvian coast. A previous study suggested that the group originated in Gondwana, and that the species arrived at their current range after the breakup of the supercontinent in the Early Cretaceous. To test this hypothesis, we infer molecular phylogenies (mitochondrial cytochrome b, ATP synthase 8/6, 12S, and 16S; nuclear rag2; total approximately 4 kb) and estimate intercontinental divergence via molecular clocks (penalized-likelihood, Bayesian relaxed clock, and universal clock rates in fishes). Age ranges for cladogenesis of African and South American lineages are 15.4-2.5 my, far more recent than would be suggested by Gondwanan vicariance; thus, the distribution of galeichthyines must be explained by dispersal or more recent vicariant events. The nested position of the Peruvian species (Galeichthys peruvianus) within the African taxa is robust, suggesting that the direction of the dispersal was from Africa to South America. The progenitor of the Peruvian species likely arrived at its current distribution with the aid of ocean currents, and several scenarios are discussed. PMID- 19187252 TI - Evolution of mammal tooth patterns: new insights from a developmental prediction model. AB - The study of mammalian evolution is often based on insights into the evolution of teeth. Developmental studies may attempt to address the mechanisms that guide evolutionary changes. One example is the new developmental model proposed by Kavanagh et al. (2007), which provides a high-level testable model to predict mammalian tooth evolution. It is constructed on an inhibitory cascade model based on a dynamic balance of activators and inhibitors, regulating differences in molar size along the lower dental row. Nevertheless, molar sizes in some mammals differ from this inhibitory cascade model, in particular in voles. The aim of this study is to point out arvicoline and murine differences within this model and to suggest an alternative model. Here we demonstrate that the inhibitory cascade is not followed, due to the arvicoline's greatly elongated first lower molar. We broaden the scope of the macroevolutionary model by projecting a time scale onto the developmental model. We demonstrate that arvicoline evolution is rather characterized by a large gap from the oldest vole to more recent genera, with the rapid acquisition of a large first lower molar contemporaneous to their radiation. Our study provides alternative evolutionary hypotheses for mammals with different trajectories of development. PMID- 19187253 TI - Relative contribution of additive, dominance, and imprinting effects to phenotypic variation in body size and growth between divergent selection lines of mice. AB - Epigenetic effects attributed to genomic imprinting are increasingly recognized as an important source of variation in quantitative traits. However, little is known about their relative contribution to phenotypic variation compared to those of additive and dominance effects, and almost nothing about their role in phenotypic evolution. Here we address these questions by investigating the relative contribution of additive, dominance, and imprinting effects of quantitative trait loci (QTL) to variation in "early" and "late" body weight in an intercross of mice selected for divergent adult body weight. We identified 18 loci on 13 chromosomes; additive effects accounted for most of the phenotypic variation throughout development, and imprinting effects were always small. Genetic effects on early weight showed more dominance, less additive, and, surprisingly, less imprinting variation than that of late weight. The predominance of additivity of QTL effects on body weight follows the expectation that additive effects account for the evolutionary divergence between selection lines. We hypothesize that the appearance of more imprinting effects on late body weight may be a consequence of divergent selection on adult body weight, which may have indirectly selected for alleles showing partial imprinting effects due to their associated additive effects, highlighting a potential role of genomic imprinting in the response to selection. PMID- 19187254 TI - Group selection on population size affects life-history patterns in the entomopathogenic nematode Steinernema carpocapsae. AB - Selection is recognized to operate on multiple levels. In disease organisms, selection among hosts is thought to provide an important counterbalance to selection for faster growth within hosts. We performed three experiments, each selecting for a divergence in group size in the entomopathogenic nematode, Steinernema carpocapsae. These nematodes infect and kill insect larvae, reproduce inside the host carcass, and emerge as infective juveniles. We imposed selection on group size by selecting among hosts for either high or low numbers of emerging nematodes. Our goal was to determine whether this trait could respond to selection at the group level, and if so, to examine what other traits would evolve as correlated responses. One of the three experiments showed a significant response to group selection. In that experiment, the high-selected treatment consistently produced more emerging nematodes per host than the low-selected treatment. In addition, nematodes were larger and they emerged later from hosts in the low-selected lines. Despite small effective population sizes, the effects of inbreeding were small in this experiment. Thus, selection among hosts can be effective, leading to both a direct evolutionary response at the population level, as well as to correlated responses in populational and individual traits. PMID- 19187255 TI - Relative role of genetic determination and plastic response during ontogeny for shell-shape traits subjected to diversifying selection. AB - We studied the relative role of genetic determination versus plastic response for traits involved in ecological adaptation of two ecotypes of Littorina saxatilis living at different shore levels. To investigate the magnitude of the plastic response across ontogeny, we compared morphological data from individuals grown in the laboratory and taken from the wild at three developmental stages: shelled embryos, juveniles, and adults. The results indicate that most shell shape variation (72-99%) in adaptive traits (globosity and aperture of the shell) is explained by the ecotype irrespective of the growth environment, suggesting that direct genetic determination is the main factor responsible for the process of adaptation in the wild. There was a tendency for the contribution of plasticity to increase over ontogeny but, in general, the direction of the plastic response did not suggest that this was adaptive. PMID- 19187256 TI - Tail shedding in island lizards [Lacertidae, Reptilia]: decline of antipredator defenses in relaxed predation environments. AB - The ability of an animal to shed its tail is a widespread antipredator strategy among lizards. The degree of expression of this defense is expected to be shaped by prevailing environmental conditions including local predation pressure. We test these hypotheses by comparing several aspects of caudal autotomy in 15 Mediterranean lizard taxa existing across a swath of mainland and island localities that differ in the number and identity of predator species present. Autotomic ease varied substantially among the study populations, in a pattern that is best explained by the presence of vipers. Neither insularity nor the presence of other types of predators explain the observed autotomy rates. Final concentration of accumulated tail muscle lactate and duration of movement of a shed tail, two traits that were previously thought to relate to predation pressure, are in general not shaped by either predator diversity or insularity. Under conditions of relaxed predation selection, an uncoupling of different aspects of caudal autotomy exists, with some elements (ease of autotomy) declining faster than others (duration of movement, lactate concentration). We compared rates of shed tails in the field against rates of laboratory autotomies conducted under standardized conditions and found very high correlation values (r > 0.96). This suggests that field autotomy rates, rather than being a metric of predatory attacks, merely reflect the innate predisposition of a taxon to shed its tail. PMID- 19187257 TI - Influence of climatic factors in the incidence of multiple sclerosis relapses in a Portuguese population. AB - BACKGROUND AND PURPOSE: Environmental factors are thought to be important in multiple sclerosis (MS) pathophysiology. We aimed to evaluate if there was an association between MS relapses and some climatic factors in a Portuguese population. METHODS: Four year retrospective study analyzing 414 MS relapses in 249 consecutive relapsing-remitting patients. Non-parametric statistics were used to compare the distribution of relapses across months and seasons. Spearman's coefficient was determined to evaluate the correlation between relapses frequency and maximum and minimum atmospheric temperatures, humidity and atmospheric pressure. RESULTS: The mean number of relapses was not significantly different between months or seasons. No correlation was found between relapse frequency and any climatic factor. CONCLUSION: Our series is one of the largest addressing the influence of specific climatic factors on MS relapses. The number of clinical MS relapses seems to be unrelated to climatic factors. PMID- 19187258 TI - Valacyclovir neurotoxicity: clinical experience and review of the literature. AB - Valacyclovir (VACV) is used increasingly to treat herpes zoster, although neuropsychiatric symptoms [VACV neurotoxicity (VAN) or acyclovir neurotoxicity], may accompany use of this drug. To promote awareness of this rare condition, we describe here two clinical cases of VAN we previously reported and review 20 cases from the literature. In all cases, chronic or acute renal failure preceded VAN. The symptoms of VAN varied, but disturbances of consciousness and hallucination occurred most commonly. When acute renal failure was due to the drug, recovery from both the disturbance of consciousness and renal failure followed within several days after discontinuation of VACV. Early recognition and diagnosis will ensure effective treatment of VAN. PMID- 19187259 TI - External shock waves therapy in dystonia: preliminary results. AB - BACKGROUND AND PURPOSE: Extracorporeal shock wave therapy (ESWT) has been shown to reduce hypertonia in patients with upper motor neuron syndrome without any side effect. The aim of the present study is to investigate whether ESWT could be useful also in patients with dystonia. METHODS: We evaluated three patients with secondary dystonia and three patients with idiopathic writer's cramp. Placebo treatment was performed in each patient. ESWT was administered during four sessions (once weekly) to the target muscles of hand and forearm using an electromagnetic lithotripter (Modulith SLK--Storz Medical). Clinical evaluation was performed using the Unified Dystonia Rating Scale in patients with secondary dystonia and the Arm Dystonia Disability Scale in patients with writer's cramp. RESULTS: After treatment, the three patients with secondary dystonia showed a marked improvement which lasted at least until 1 month after the last session. In the patients with writer's cramp, the improvement after ESWT was less consistent being effective only in two subjects. There were no associated adverse effects. CONCLUSIONS: Extracorporeal shock wave therapy is probably an effective and safe treatment for upper limb dystonia, particularly for the secondary forms. Larger randomized studies are needed to confirm these preliminary results. PMID- 19187260 TI - The 'hot cross bun' sign in the patients with spinocerebellar ataxia. AB - BACKGROUND AND PURPOSE: The 'hot cross bun' sign (HCBS), typically seen in the patients with multiple system atrophy, refers to a cruciform hyperintensity in the pons on T2-weighted MRI. Little is known about its pathological basis and prevalence in other degenerative cerebellar diseases and healthy population. We investigate the frequency of HCBS in the patients with spinocerebellar ataxia (SCA) and healthy controls. METHODS: The presence of HCBS on T2-weighted axial MRIs from 138 SCA patients (three SCA1, 35 SCA2, 76 SCA3, 18 SCA6, one SCA7, three SCA8, and two SCA17) and 102 healthy controls was evaluated retrospectively. RESULTS: The overall prevalence of HCBS in the SCA patients is 8.7%, but the frequency varies in different subtypes: 25.7% in SCA2, 1.3% in SCA3, and none in SCA6 or healthy controls. Notably, one patient with SCA7 and one with SCA8 were also found to have HCBS. CONCLUSIONS: The differential list of HCBS should be expanded to include SCA7 and SCA8. The elucidation of frequency of HCBS in various SCA subtypes may help prioritize the genetic testing in late onset dominant ataxia. PMID- 19187261 TI - Natalizumab is effective as second line therapy in the treatment of relapsing remitting multiple sclerosis. AB - BACKGROUND: Natalizumab has been recommended for the treatment of patients with relapsing remitting multiple sclerosis with insufficient response to interferon beta (IFN-beta) or glatiramer acetate (GA). METHOD: Prospective, observational study. RESULTS: We found a reduction of the annualized relapse rate from 2.1 under IFN-beta or GA to 0.2 one year after switching to natalizumab. There were 94% fewer gadolinium enhancing lesions with natalizumab. CONCLUSION: Natalizumab reduced short term clinical and MRI activity in second line therapy and efficacy is comparable to first line therapy as demonstrated in the pivotal trials. PMID- 19187262 TI - The effect of onset age on the clinical features of Parkinson's disease. AB - Many clinicians view age at onset as an important determinant of clinical phenotype in Parkinson's disease (PD) and this has been reinforced by the identification of Mendelian genes that account for some cases of younger onset PD. A systematic review of OVID Medline for articles relevant to the relationship between clinical features and age at onset in PD published in English between 1950-2007 was performed. There are very few prospective community based studies which focus on the relationship between age at onset and the features of PD and a variety of case definitions are used in the literature. Most studies of young onset PD are based on specialist clinic referral series. The available evidence suggests that PD patients with a younger age at onset have: (i) a slower disease progression, (ii) an increased rate of dystonia at onset and during treatment, (iii) a lower rate of dementia and (iv) an increased rate of dyskinesias in response to L-DOPA treatment. The majority of the available studies do not report patient genotype data, but it is probably that the clinical heterogeneity of PD will be further refined with detailed clinico-genetic studies. PMID- 19187263 TI - A decrease of human leucocyte antigen-DR expression on monocytes in peripheral blood predicts stroke-associated infection in critically-ill patients with acute stroke. AB - BACKGROUND AND PURPOSE: To investigate changes in human leucocyte antigen (HLA) DR expression on peripheral monocytes, determine the value of predicting the development of stroke-associated infection (SAI), and determine the correlation with other conditions in critically-ill patients in the neurological intensive care unit (NICU) who suffered an acute stroke. METHODS: All patients were enrolled consecutively and admitted to NICU within 24 h after the onset of symptoms. Patients were followed in order to identify whether infection developed and determine survival status within 2 weeks after the stroke. Patients were divided into stroke or control group by study design, infection or non-infection group by whether or not they had an infection, survival or death group by prognosis and cerebral infarction or cerebral haemorrhage group by stroke type. Patients in which acute stroke was excluded by head CT or MRI were admitted to general ward and were used as a control group. Blood samples were collected serially on days 1, 2, 4, 6 and 14 after stroke, then monocyte human leucocyte antigen-DR (HLA-DR) expression was determined by flow cytometry. The National Institute of Health Stroke Scale (NIHSS), Acute Physiology and Chronic Health Evaluation II (APACHEII) and Glasgow Coma Scale (GCS) scores were recorded over the course of observation. RESULTS: Fifty-three subjects and 39 controls were enrolled in the study. On days 1, 2, 4, 6 and 14, there was a significant difference in monocyte HLA-DR expression between stroke group and control group (all P < 0.001), but no difference was found between ischaemic stroke group and haemorrhagic stroke group (all P > 0.05). The infection group compared with non infection group did not exhibit a significant difference in HLA-DR expression on days 1 and 2 (all P > 0.05), but significant differences emerged on days 4, 6 and 14 (all P < 0.01). On days 1 and 2 the HLA-DR expression in the survival group compared with death group, was not significantly different (all P > 0.05), but differences became significant on days 4 and 6 (P < 0.01). On day 1, HLA-DR expression <62.80% had the predictive value to SAI (sensitivity 83.3%, specificity 55.2%, AUC = 0.661, P = 0.031) and on day 2, HLA-DR expression <57.83% had the predictive value to SAI (sensitivity 95.8%, specificity 79.3%, AUC = 0.907, P = 0.000) in acute stroke patients. A statistically significant inverse correlation was found between NIHSS and HLA-DR on days 2 and 4 during the observation period (all P < 0.01), but there was no statistically significant negative correlation on days 1, 6 or 14 (all P > 0.05). HLA-DR expression did not correlate with APACHEII (all P > 0.05) or GCS (all P > 0.05) during the measurement period. CONCLUSIONS: Human leucocyte antigen-DR (HLA-DR) expression decreases and sustains a dynamic change and it also relates to the severity of patient's condition in the critically-ill patients with stroke. Progressively persistent low monocyte HLA-DR expression is associated with a poor prognosis. The decline in HLA-DR expression contributes to infection in critically-acute stroke patients. Monitoring of monocyte HLA-DR expression may be useful for identifying patients suffering from acute stroke who are at high risk for infection. PMID- 19187264 TI - Asymmetrical lateral ventricular enlargement in Parkinson's disease. AB - BACKGROUND: A recent case report suggested the presence of asymmetrical lateral ventricular enlargement associated with motor asymmetry in Parkinson's disease (PD). The current study explored these associations further. METHODS: Magnetic resonance imaging (3T) scans were obtained on 17 PD and 15 healthy control subjects at baseline and 12-43 months later. Baseline and longitudinal lateral ventricular volumetric changes were compared between contralateral and ipsilateral ventricles in PD subjects relative to symptom onset side and in controls relative to their dominant hand. Correlations between changes in ventricular volume and United Parkinson's disease rating scale motor scores (UPDRS-III) whilst on medication were determined. RESULTS: The lateral ventricle contralateral to symptom onset side displayed a faster rate of enlargement compared to the ipsilateral (P = 0.004) in PD subjects, with no such asymmetry detected (P = 0.312) in controls. There was a positive correlation between ventricular enlargement and worsening motor function assessed by UPDRS-III scores (r = 0.96, P < 0.001). DISCUSSION: There is asymmetrical lateral ventricular enlargement that is associated with PD motor asymmetry and progression. Further studies are warranted to investigate the underlying mechanism(s), as well as the potential of using volumetric measurements as a marker for PD progression. PMID- 19187266 TI - alpha7 and non-alpha7 nicotinic acetylcholine receptors modulate dopamine release in vitro and in vivo in the rat prefrontal cortex. AB - Nicotine enhances attentional and working memory aspects of executive function in the prefrontal cortex (PFC) where dopamine plays a major role. Here, we have determined the nicotinic acetylcholine receptor (nAChR) subtypes that can modulate dopamine release in rat PFC using subtype-selective drugs. Nicotine and 5-Iodo-A-85380 (beta2* selective) elicited [(3)H]dopamine release from both PFC and striatal prisms in vitro and dopamine overflow from medial PFC in vivo. Blockade by dihydro-beta-erythroidine supports the participation of beta2* nAChRs. However, insensitivity of nicotine-evoked [(3)H]dopamine release to alpha conotoxin-MII in PFC prisms suggests no involvement of alpha6beta2* nAChRs, in contrast to the striatum, and this distinction is supported by immunoprecipitation of nAChR subunits from these tissues. The alpha7 nAChR selective agonists choline and Compound A also promoted dopamine release from PFC in vitro and in vivo, and their effects were enhanced by the alpha7 nAChR selective allosteric potentiator PNU-120596 and blocked by specific antagonists. DNQX and MK801 inhibited [(3)H]dopamine release evoked by choline and PNU-120596, suggesting crosstalk between alpha7 nAChRs, glutamate and dopamine in the PFC. In vivo, systemic (but not local) administration of PNU-120596, in the absence of agonist, facilitated dopamine overflow in the medial PFC, consistent with the activation of extracortical alpha7 nAChRs by endogenous acetylcholine or choline. These data establish that both beta2* and alpha7 nAChRs can modulate dopamine release in the PFC in vitro and in vivo. Through their distinct actions on dopamine release, these nAChR subtypes could contribute to executive function, making them specific therapeutic targets for conditions such as schizophrenia and attention deficit hyperactivity disorder. PMID- 19187265 TI - A direct main olfactory bulb projection to the 'vomeronasal' amygdala in female mice selectively responds to volatile pheromones from males. AB - The main olfactory system, like the accessory olfactory system, responds to pheromones involved in social communication. Whereas pheromones detected by the accessory system are transmitted to the hypothalamus via the medial ('vomeronasal') amygdala, the pathway by which pheromones are detected and transmitted by the main system is not well understood. We examined in female mice whether a direct projection from mitral/tufted (M/T) cells in the main olfactory bulb (MOB) to the medial amygdala exists, and whether medial amygdala-projecting M/T cells are activated by volatile urinary odors from conspecifics or a predator (cat). Simultaneous anterograde tracing using Phaseolus vulgaris leucoagglutinin and Fluoro-Ruby placed in the MOB and accessory olfactory bulb (AOB), respectively, revealed that axons of MOB M/T cells projected to superficial laminae of layer Ia in anterior and posterodorsal subdivisions of the medial amygdala, whereas projection neurons from the AOB sent axons to non-overlapping, deeper layer Ia laminae of the same subdivisions. Placement of the retrograde tracer cholera toxin B into the medial amygdala labeled M/T cells that were concentrated in the ventral MOB. Urinary volatiles from male mice, but not from female conspecifics or cat, induced Fos in medial amygdala-projecting MOB M/T cells of female subjects, suggesting that information about male odors is transmitted directly from the MOB to the 'vomeronasal' amygdala. The presence of a direct MOB-to-medial amygdala pathway in mice and other mammals could enable volatile, opposite-sex pheromones to gain privileged access to diencephalic structures that control mate recognition and reproduction. PMID- 19187267 TI - Increase in presynaptic territory of C-terminals on lumbar motoneurons of G93A SOD1 mice during disease progression. AB - Compensatory synaptic plasticity is reported in muscle and the central nervous system of motor neuron disease patients, and transgenic SOD1 mice, but direct ultrastructural evidence for spinal motoneurons is lacking. Prompted by our observation in spinal cords from autopsied patients suggesting selective enlargement of the ultrastructurally distinctive C-type terminal synapsing with spinal motoneurons, we examined the ultrastructural synaptology of lumbar motoneurons during disease progression in age- and sex-matched wild-type mice, transgenic G93A SOD1 mice, and mice overexpressing normal human SOD1 (Wt(SOD1)). Prescribed criteria classified presynaptic terminals of motoneurons into five ultrastructural classes (S, F, T, M, and C). Computerized morphometry on electronmicrographs was used to measure their appositional lengths, coverage of the motoneuron membrane, and sizes of postsynaptic structures. No terminal degeneration occurred in wild-type or Wt(SOD1) mice. In transgenic mice, degeneration of motoneurons and S-terminals and F-terminals commenced presymptomatically (10 weeks), and continued into the symptomatic stage (18 weeks). However, C-terminals were preserved. Morphometry confirmed significant reductions in frequency and membrane coverage for S-terminals and F-terminals between 10 and 18 weeks, but a maintained frequency of C-terminals coupled with increased appositional length and coverage of the motoneuron membrane. Increased C-terminal size was matched by growth of its characterizing postsynaptic cistern and Nissl body. The results reveal selective preservation and increased presynaptic territory of the C-type terminal. As C-terminals derive from cholinergic intrasegmental propriospinal interneurons and may modulate motoneuron excitability, their increased presynaptic territory on surviving motoneurons of transgenic SOD1 mice may represent a means of maintaining excitability, compensating for the loss of overall presynaptic input. PMID- 19187268 TI - Characterisation of tremor-associated local field potentials in the subthalamic nucleus in Parkinson's disease. AB - We simultaneously recorded local field potentials (LFPs) in the subthalamic nucleus (STN) and surface electromyographic signals (EMGs) from the extensor and flexor muscles of the contralateral forearm in eight patients with idiopathic tremor-dominant Parkinson's disease (resting tremor) during the bilateral implantation of deep brain stimulation electrodes. Recordings were made at different heights (in 0.5- to 2.0-mm steps beginning outside the STN) using up to five concentrically configured macroelectrodes (2 mm apart). The patients were instructed to relax their contralateral forearm (rest condition). We analysed the coherence between tremor EMGs and STN LFPs, which showed significant tremor associated coupling at single tremor and double tremor frequencies. Moreover, the EMG-LFP coherences were characterised by differences between antagonistic muscles (flexor, extensor) and by the spatial distribution of LFPs within the STN. Coherence at single and double tremor frequencies occurred significantly more frequently within STN than above STN (in the zona incerta). In this study, we were able to show that, within STN, tremor-associated LFP activity varied with spatial distribution and with the contralateral antagonistic forearm muscles. These findings suggest the existence of distribution- and muscle-specific tremor associated LFP activity at different tremor frequencies and an organisation of tremor-related subloops within the STN. PMID- 19187269 TI - Astrocyte-derived kynurenic acid modulates basal and evoked cortical acetylcholine release. AB - We tested the hypothesis that fluctuations in the levels of kynurenic acid (KYNA), an endogenous antagonist of the alpha7 nicotinic acetylcholine (ACh) receptor, modulate extracellular ACh levels in the medial prefrontal cortex in rats. Decreases in cortical KYNA levels were achieved by local perfusion of S ESBA, a selective inhibitor of the astrocytic enzyme kynurenine aminotransferase II (KAT II), which catalyses the formation of KYNA from its precursor L kynurenine. At 5 mm, S-ESBA caused a 30% reduction in extracellular KYNA levels, which was accompanied by a two-threefold increase in basal cortical ACh levels. Co-perfusion of KYNA in the endogenous range (100 nm), which by itself tended to reduce basal ACh levels, blocked the ability of S-ESBA to raise extracellular ACh levels. KYNA perfusion (100 nm) also prevented the evoked ACh release caused by d amphetamine (2.0 mg/kg). This effect was duplicated by the systemic administration of kynurenine (50 mg/kg), which resulted in a significant increase in cortical KYNA formation. Jointly, these data indicate that astrocytes, by producing and releasing KYNA, have the ability to modulate cortical cholinergic neurotransmission under both basal and stimulated conditions. As cortical KYNA levels are elevated in individuals with schizophrenia, and in light of the established role of cortical ACh in executive functions, our findings suggest that drugs capable of attenuating the production of KYNA may be of benefit in the treatment of cognitive deficits in schizophrenia. PMID- 19187271 TI - High intracellular content of cyclin-dependent kinase inhibitor p27(Kip1) in early- and intermediate stage B-cell chronic lymphocytic leukemia lymphocytes predicts rapid progression of the disease. AB - Previous studies showed that peripheral blood lymphocytes of B-cell chronic lymphocytic leukemia (B-CLL) displayed a high intracellular level of cell cycle inhibitory protein p27(Kip1). It has been suggested that its' high expression may confer them survival advantage and lead to unfavorable prognosis, but the prognostic significance of p27(Kip1) expression for previously untreated, non advanced stage B-CLL was not established. We studied a relationship between the intracellular level of p27(Kip1) of lymphocytes of early- and intermediate stage B-CLL patients and their spontaneous apoptosis in vitro, as well as prognostic significance of p27(Kip1) in B-CLL lymphocytes for the risk of disease progression. Intracellular p27(Kip1) content of peripheral blood lymphocytes obtained from 48 previously untreated 0-II Rai stage B-CLL patients was determined by flow cytometry. The viability and apoptosis of those lymphocytes after 72-h culture were also assessed. During the follow-up period (6-71 months, median 59.5), we recorded the time elapsed to the doubling of lymphocyte count, progression to a higher Rai stage and the appearance of indications for cytostatic treatment. The p27(Kip1) expression was neither correlated with initial lymphocyte count, CD38 expression, cell viability nor spontaneous apoptosis ratio after 72-h culture. Higher p27(Kip1) level was related to the probability of earlier occurrence of each of three above-mentioned events. We did not find a prognostic significance of in vitro cell viability nor apoptosis as to the risk of disease progression. Our results indicate that elevated intracellular p27(Kip1) level in leukemic lymphocytes of early- and intermediate stage B-CLL patients contributes to rapid progression of the disease. PMID- 19187272 TI - Pyoderma gangrenosum as initial presentation of chronic myelomonocytic leukemia. PMID- 19187270 TI - c-Met signaling promotes IL-6-induced myeloma cell proliferation. AB - OBJECTIVES: Hepatocyte growth factor (HGF) is a constituent of the myeloma microenvironment and is elevated in sera from myeloma patients compared to healthy individuals. Increased levels of serum HGF predict a poor prognosis. It has previously been shown by us and others HGF can act as a growth factor to myeloma cells in vitro although these effects have been moderate. We therefore wanted to investigate if HGF could influence the effects of interleukin (IL)-6. METHODS: Myeloma cell lines and primary samples were tested for the combined effects of IL-6 and HGF in inducing DNA synthesis and migration. Expression levels of c-Met protein were analysed by Western blotting and flow cytometry. Signaling pathways were examined by Western blotting using phosphospecific antibodies and a Ras-GTP pull down assay. RESULTS: HGF potentiated IL-6-induced growth in human myeloma cell lines and in purified primary myeloma cells. There was also cooperation between HGF and IL-6 in induction of migration. There seemed to be two explanations for this synergy. IL-6-treatment increased the expression of c-Met making cells HGF responsive, and IL-6 was dependent on c-Met signaling in activating both Ras and p44/42 MAPK by a mechanism involving the tyrosine phosphatase Shp2. CONCLUSIONS: The results indicate that besides from being a myeloma growth factor alone, HGF can also potentiate the effects of IL-6 in myeloma proliferation and migration. Thus, c-Met signaling could be a target for therapy of multiple myeloma. PMID- 19187273 TI - B-Lymphocyte stimulator: a new biomarker for multiple myeloma. AB - Multiple myeloma (MM) is a common malignant tumor, characterized by unlimited proliferation of abnormal plasmocytes in bone marrow. Considering the biological function of B-Lymphocyte stimulator (BLyS) and its receptors in B cell, we examined BLyS and its receptors expression in MM cells. Our studies confirmed that BLyS and its receptors are expressed in MM cells, including KM3, CZ-1, and primary MM cells, playing an important role in the survival and proliferation of MM cells. Additionally, we provide evidence that BLyS protein is located in the MM cell plasma membrane. We also found that IFN-gamma and IL-6 can induce BLyS expression on MM cells, while after the treatment of BAY11-7082, an IkB-alpha phosphorylation inhibitor, IFN-gamma induced up regulation of BLyS was completely inhibited, suggesting that nuclear factor kappaB (NF-kappaB) might be involved in the mechanism of the regulation of BLyS levels in response to cytokines. Finally, linear correlation analysis of the Lactate Dehydrogenase concentration and beta 2 microglobulin level with BLyS, and expressions of BLyS mRNA in MM patients revealed a significant correlation between them (P < 0.01 in all case), showing that BLyS could be a biomarker for the diagnosis and treatment of MM. PMID- 19187274 TI - Platelet satellitism and lympho-agglutination as presenting finding in marginal zone B-cell lymphoma. PMID- 19187275 TI - Plasma-based detection of clonality in lymphoid malignancies. AB - OBJECTIVES: Plasma has been found to be enriched with tumor-specific DNA, RNA, and protein in patients with hematologic disease. We assessed the utility of plasma as a DNA source for detection of genetic abnormalities in patients with suspected B- or T-cell lymphoproliferative disorders. METHODS: DNA was extracted from paired peripheral blood (PB) cells and plasma for polymerase chain reaction (PCR)-based detection of immunoglobulin heavy chain (IgH) and T-cell receptor gamma chain (TCR-gamma) rearrangements, and B-cell leukemia/lymphoma (BCL)-1/IgH and BCL-2/IgH translocations. RESULTS: Concordance between plasma and PB cell analysis was 100% for IgH (n = 57), TCR-gamma (n = 57), and BCL-1/IgH (n = 37) rearrangements, and 94% (60/64) for BCL-2/IgH; four of 11 plasma samples positive for BCL-2/IgH tested negative in paired cells. No plasma or PB cell samples from 195 healthy donors showed genetic abnormalities. CONCLUSIONS: These findings indicate that plasma is a reliable sample type for detection of abnormalities associated with B- and T-cell lymphoproliferative disorders, providing sensitivity equal to or greater than that of PB cells. PMID- 19187276 TI - Rivaroxaban -- an oral, direct Factor Xa inhibitor: lessons from a broad clinical study programme. AB - Anticoagulants are recommended for the prevention and treatment of venous thromboembolism (VTE), prevention of stroke in patients with atrial fibrillation (AF) and secondary prevention in patients with acute coronary syndrome (ACS). There is a clinical need for novel anticoagulants offering improvements over current standard of care, such as fixed oral dosing and no need for routine monitoring. Rivaroxaban, an oral, once-daily, direct Factor Xa inhibitor, has recently completed the RECORD phase III programme for the prevention of VTE in patients undergoing total hip or knee replacement (THR or TKR), an indication for which it is approved in Europe and Canada. It is being investigated in large scale phase III studies for VTE treatment and prevention of stroke in patients with AF, and phase III studies will soon commence for secondary prevention in patients with ACS. Phase I studies demonstrated that no routine anticoagulation monitoring was required, while phase II studies suggested that fixed daily doses had a wide therapeutic window. The four RECORD studies consistently showed that rivaroxaban was significantly more effective than enoxaparin in the prevention of VTE after THR and TKR, with a similar safety profile. This review describes the development of this novel anticoagulant, from bench to bedside. PMID- 19187277 TI - The role of Th17 cells in adult patients with chronic idiopathic thrombocytopenic purpura. PMID- 19187278 TI - Efficacy and safety of deferasirox, an oral iron chelator, in heavily iron overloaded patients with beta-thalassaemia: the ESCALATOR study. AB - OBJECTIVE: Many patients with transfusional iron overload are at risk for progressive organ dysfunction and early death and poor compliance with older chelation therapies is believed to be a major contributing factor. Phase II/III studies have shown that oral deferasirox 20-30 mg/kg/d reduces iron burden, depending on transfusional iron intake. METHODS: The prospective, open-label, 1 yr ESCALATOR study in the Middle East was designed to evaluate once-daily deferasirox in patients > or =2 yr with beta-thalassaemia major and iron overload who were previously chelated with deferoxamine and/or deferiprone. Most patients began treatment with deferasirox 20 mg/kg/d; doses were adjusted in response to markers of over- or under-chelation. The primary endpoint was treatment success, defined as a reduction in liver iron concentration (LIC) of > or =3 mg Fe/g dry weight (dw) if baseline LIC was > or =10 mg Fe/g dw, or final LIC of 1-7 mg Fe/g dw for patients with baseline LIC of 2 to <10 mg Fe/g dw. RESULTS: Overall, 233/237 enrolled patients completed 1 yr's treatment. Mean baseline LIC was 18.0 +/- 9.1 mg Fe/g dw, while median serum ferritin was 3356 ng/mL. After 1 yr's deferasirox treatment, the intent-to-treat population experienced a significant treatment success rate of 57.0% (P = 0.016) and a mean reduction in LIC of 3.4 mg Fe/g dw. Changes in serum ferritin appeared to parallel dose increases at around 24 wk. Most patients (78.1%) underwent dose increases above 20 mg/kg/d, primarily to 30 mg/kg/d. Drug-related adverse events were mostly mild to moderate and resolved without discontinuing treatment. CONCLUSIONS: The results of the ESCALATOR study in primarily heavily iron-overloaded patients confirm previous observations in patients with beta-thalassaemia, highlighting the importance of timely deferasirox dose adjustments based on serum ferritin levels and transfusional iron intake to ensure patients achieve their therapeutic goal of maintenance or reduction in iron burden. PMID- 19187279 TI - Diagnostic value of zinc protoporphyrin in a screening strategy for alpha thalassemia. AB - The definitive diagnosis of alpha-thalassemia involves detection of a deletion of one or more alpha-globin that encode the alpha-chains of Hb (hemoglobin). To determine whether DNA analysis is indicated, screening tests such as mean corpuscular volume (MCV) and Hb typing are employed. alpha-Thalassemia often correlates with normal or low HbA2 values. Zinc protoporphyrin (ZPP) is usually high in ferropenic anemia or lead-poisoning and is normal or slightly raised in beta-thalassemia. Therefore, ZPP is currently used as a marker to discriminate between ferropenic anemia and beta-thalassemia. We investigated the diagnostic potential of ZPP < 150 micromol/mol heme in a screening strategy for alpha thalassemia. We measured ZPP and performed DNA analysis for detecting the seven most prevalent alpha-thalassemia deletions, namely, alpha3.7, SEA, alpha20.5, alpha4.2, MED, FIL, and THAI, in the blood samples of 200 patients with MCV < 70 fL and HbA2 < or = 3.5%. Deletions were detected in 9% subjects in the ZPP > or = 150 group (n = 175) and 56% subjects in the ZPP < 150 group (n = 29); this difference was statistically significant (chi-square test, P < 0.001). We conclude that ZPP < 150 micromol/mol heme can be used in a new screening strategy for alpha-thalassemia. PMID- 19187280 TI - Lennox-Gastaut syndrome with late-onset and prominent reflex seizures in trisomy 21 patients. AB - PURPOSE: Lennox-Gastaut syndrome (LGS) is a severe epileptic condition characterized by multiple seizure types including tonic seizures, slow spike-and wave discharges on electroencephalography (EEG), and cognitive impairment. LGS can occur in apparently healthy subjects or in patients with preexisting brain damage. The onset peaks between 3 and 5 years of age and the prognosis is usually poor. Herein we report 13 subjects with trisomy 21 who developed LGS. METHODS: We retrospectively reviewed the clinical and EEG data of consecutive patients with LGS and trisomy 21 referred to five epilepsy centers over the last 30 years. RESULTS: Data for 13 patients (8 male, 5 female) were collected. The mean age at onset was 9.1 years (range 5-16). The mean age at last follow-up was 23.5 years (range 11-43 years). Seizure onset was after age 8 years in eight (62%) patients and between age 5 and 8 in the other five. In none of the cases did a West syndrome precede the onset of LGS. Nine of 13 patients (69%) had unambiguous reflex seizures, mostly precipitated by sudden unexpected sensory stimulations, usually preceding or accompanying the onset of a full-blown LGS picture. Interictal and ictal EEG findings were typical for LGS. All patients were drug resistant. DISCUSSION: Patients with trisomy 21 may present a peculiar LGS, characterized by late onset and high occurrence of reflex seizures. Mechanisms underlying this particular presentation of LGS may include dendritic rarefaction and decreased interneurons, as well as functional abnormalities leading to overall decreased brain inhibition in these patients. PMID- 19187281 TI - Soil CO2 flux and photoautotrophic community composition in high-elevation, 'barren' soil. AB - Soil-dominated ecosystems, with little or no plant cover (i.e. deserts, polar regions, high-elevation areas and zones of glacial retreat), are often described as 'barren', despite their potential to host photoautotrophic microbial communities. In high-elevation, subnival zone soil (i.e. elevations higher than the zone of continuous vegetation), the structure and function of these photoautotrophic microbial communities remains essentially unknown. We measured soil CO(2) flux at three sites (above 3600 m) and used molecular techniques to determine the composition and distribution of soil photoautotrophs in the Colorado Front Range. Soil CO(2) flux data from 2002 and 2007 indicate that light driven CO(2) uptake occurred on most dates. A diverse community of Cyanobacteria, Chloroflexi and eukaryotic algae was present in the top 2 cm of the soil, whereas these clades were nearly absent in deeper soils (2-4 cm). Cyanobacterial communities were composed of lineages most closely related to Microcoleus vaginatus and Phormidium murrayi, eukaryotic photoautotrophs were dominated by green algae, and three novel clades of Chloroflexi were also abundant in the surface soil. During the light hours of the 2007 snow-free measurement period, CO(2) uptake was conservatively estimated to be 23.7 g C m(-2) season(-1). Our study reveals that photoautotrophic microbial communities play an important role in the biogeochemical cycling of subnival zone soil. PMID- 19187282 TI - Occurrence of phosphate acquisition genes in Prochlorococcus cells from different ocean regions. AB - The cyanobacterium Prochlorococcus is the numerically dominant phototroph in oligotrophic parts of the oceans. Recently, it was shown that the distribution of phosphate acquisition genes did not match the 16S rRNA phylogeny among isolates from this group but rather appeared related to phosphate availability where the strains had been isolated. To further understand adaptation to phosphate limitation in Prochlorococcus, the distribution of phosphate acquisition genes was investigated in different ocean regions and related to local ortho-phosphate concentration. In regions characterized by less than 0.1 microM phosphate, most Prochlorococcus cells contain genes involved in phosphate uptake, regulation and utilization of organic phosphates. In contrast, most of these genes are absent in regions with more than 0.1 microM phosphate with the exception of genes involved in transport of phosphate (phoE and pstABCS) and three genes of unknown function. This pattern of phosphate acquisition genes showed no significant correspondence to the distribution of rRNA phylotypes. In addition, it was demonstrated that several genes in a separate genomic island were commonly present in low-P sites while absent in high-P sites. Overall, this study further demonstrates a linkage between environmental conditions in the ocean and genome content of Prochlorococcus. PMID- 19187283 TI - Genome sequence of Desulfobacterium autotrophicum HRM2, a marine sulfate reducer oxidizing organic carbon completely to carbon dioxide. AB - Sulfate-reducing bacteria (SRB) belonging to the metabolically versatile Desulfobacteriaceae are abundant in marine sediments and contribute to the global carbon cycle by complete oxidation of organic compounds. Desulfobacterium autotrophicum HRM2 is the first member of this ecophysiologically important group with a now available genome sequence. With 5.6 megabasepairs (Mbp) the genome of Db. autotrophicum HRM2 is about 2 Mbp larger than the sequenced genomes of other sulfate reducers (SRB). A high number of genome plasticity elements (> 100 transposon-related genes), several regions of GC discontinuity and a high number of repetitive elements (132 paralogous genes Mbp(-1)) point to a different genome evolution when comparing with Desulfovibrio spp. The metabolic versatility of Db. autotrophicum HRM2 is reflected in the presence of genes for the degradation of a variety of organic compounds including long-chain fatty acids and for the Wood Ljungdahl pathway, which enables the organism to completely oxidize acetyl-CoA to CO(2) but also to grow chemolithoautotrophically. The presence of more than 250 proteins of the sensory/regulatory protein families should enable Db. autotrophicum HRM2 to efficiently adapt to changing environmental conditions. Genes encoding periplasmic or cytoplasmic hydrogenases and formate dehydrogenases have been detected as well as genes for the transmembrane TpII-c(3), Hme and Rnf complexes. Genes for subunits A, B, C and D as well as for the proposed novel subunits L and F of the heterodisulfide reductases are present. This enzyme is involved in energy conservation in methanoarchaea and it is speculated that it exhibits a similar function in the process of dissimilatory sulfate reduction in Db. autotrophicum HRM2. PMID- 19187285 TI - Response to the paper by Sara Pantuliano. PMID- 19187284 TI - Synergistic role of curli and cellulose in cell adherence and biofilm formation of attaching and effacing Escherichia coli and identification of Fis as a negative regulator of curli. AB - Curli are adhesive fimbriae of Escherichia coli and Salmonella enterica. Expression of curli (csgA) and cellulose (bcsA) is co-activated by the transcriptional activator CsgD. In this study, we investigated the contribution of curli and cellulose to the adhesive properties of enterohaemorragic (EHEC) O157:H7 and enteropathogenic E. coli (EPEC) O127:H6. While single mutations in csgA, csgD or bcsA in EPEC and EHEC had no dramatic effect on cell adherence, double csgAbcsA mutants were significantly less adherent than the single mutants or wild-type strains to human colonic HT-29 epithelial cells or to cow colon tissue in vitro. Overexpression of csgD (carried on plasmid pCP994) in a csgD mutant, but not in the single csgA or bscA mutants, led to significant increase in adherence and biofilm formation in EPEC and EHEC, suggesting that synchronized over-production of curli and cellulose enhances bacterial adherence. In line with this finding, csgD transcription was activated significantly in the presence of cultured epithelial cells as compared with growth in tissue culture medium. Analysis of the influence of virulence and global regulators in the production of curli in EPEC identified Fis (factor for inversion stimulation) as a, heretofore unrecognized, negative transcriptional regulator of csgA expression. An EPEC E2348/69Deltafis produced abundant amounts of curli whereas a double fis/csgD mutant yielded no detectable curli production. Our data suggest that curli and cellulose act in concert to favour host colonization, biofilm formation and survival in different environments. PMID- 19187286 TI - Safety, tolerability, pharmacodynamics and pharmacokinetics of albiglutide, a long-acting glucagon-like peptide-1 mimetic, in healthy subjects. AB - AIMS: Albiglutide is a glucagon-like peptide-1 (GLP-1) mimetic generated by genetic fusion of a dipeptidyl peptidase-IV-resistant GLP-1 dimer to human albumin. Albiglutide was designed to retain the therapeutic effects of native GLP 1 while extending its duration of action. This study was conducted to determine the pharmacokinetics and initial safety/tolerability profile of albiglutide in non-diabetic volunteers. METHODS: In this single-blind, randomized, placebo controlled trial, 39 subjects (18-60 years, body mass index 19.9-35.0 kg/m(2)) received placebo (n = 10) or escalating doses of albiglutide (n = 29) on days 1 and 8 in the following sequential cohorts: cohort 1: 0.25 + 1 mg; cohort 2: 3 + 6 mg; cohort 3: 16 + 24 mg; cohort 4: 48 + 60 mg; and cohort 5: 80 + 104 mg. Dose proportionality was evaluated based on area under the plasma drug concentration versus time curve [area under the curve (AUC((0-7 days)))] and maximum plasma drug concentration (C(max)) for cohorts 2-5 during week 1. RESULTS: Albiglutide had a terminal elimination half-life (T(1/2)) of 6-8 days and time to maximum observed plasma drug concentration (T(max)) of 3-4 days. A greater-than-dose proportional increase in albiglutide exposure was observed. Albiglutide demonstrated a dose-dependent trend in reductions of glucose weighted mean AUC and fructosamine levels in healthy subjects. The incidence and severity of adverse events (AEs) was similar between placebo and albiglutide groups. Headache was the most frequent drug-related AE, followed by constipation, flatulence and nausea. CONCLUSIONS: Albiglutide has a half-life that favours once weekly or less frequent dosing with an acceptable safety/tolerability profile in non-diabetic subjects. PMID- 19187287 TI - Introduction. Posttraumatic epilepsy: treatable epileptogenesis. PMID- 19187288 TI - Epilepsy after head injury: an overview. AB - Traumatic brain injury (TBI) has been recognized as a cause of epilepsy since antiquity, and it remains one of the most common and important causes of acquired epilepsy today. Epidemiologic studies have demonstrated a clear relationship between the severity of injury and the likelihood of developing epilepsy, with the risk approaching 50% in TBI cases associated with direct injury to brain parenchyma. Importantly, many TBI victims develop epilepsy months or years following the initial injury, making this patient population a prime target for the development of antiepileptogenesis therapies. However, progress in this area of clinical research is hindered by the lack of reliable and valid biomarkers. Given current events in the Middle East and elsewhere, the importance of TBI and epilepsy deserves special attention due to the increase in severe head trauma associated with modern warfare. PMID- 19187289 TI - Preventing and treating posttraumatic seizures: the human experience. AB - Posttraumatic epilepsy presents an ideal target for prevention efforts. Traumatic brain injury (TBI) is common, characteristics that put people at high risk such as penetrating injury or subdural hematoma or provoked seizures are easily identified, and the latency between the injury and the onset of epileptic seizures is frequently short. Several drugs have been tested for their ability to prevent provoked seizures and epilepsy after TBI. We describe the design of those studies and their results. Phenytoin and carbamazepine significantly reduce the incidence of provoked seizures. Phenobarbital and the combination of phenobarbital and phenytoin also look promising for reducing provoked seizures, but small sample sizes in the studies evaluating these drugs do not allow definitive conclusions. None of the drugs studied (phenytoin, phenobarbital, their combination, carbamazepine, valproate, or magnesium) have shown reliable evidence that they prevent, or even suppress, epileptic seizures after TBI. For most of the regimens tested (the phenytoin/phenobarbital combination being the exception), the best estimate of effect is under a 25% reduction in posttraumatic seizures, well less than the 50% reduction most studies were designed to detect. The evaluation of the tested drugs has serious limitations, however, and antiepileptic drugs (AEDs) developed since 1980 and other compounds have barely been tested at all. Better understanding the process of epileptogenesis, testing treatments that demonstrate antiepileptogenic effects in the laboratory, and performing thorough preclinical and phase II evaluations before attempting definitive trials should greatly improve the chance of identifying ways to prevent posttraumatic epilepsy, providing the ultimate cure for this condition. PMID- 19187290 TI - Posttraumatic epilepsy: the endophenotypes of a human model of epileptogenesis. AB - Posttraumatic epilepsy is a common complication of traumatic brain injury (TBI), occurring in up to 15-20% of patients with severe brain trauma. Trauma accounts for approximately 5% of chronic epilepsy in the community. Because it is a common condition, and because of the relatively short latency period between injury and onset of chronic seizures, posttraumatic epilepsy represents a good model to test antiepileptogenic therapies. However, several well-conducted clinical trials have failed to demonstrate antiepileptogenic efficacy for several common anticonvulsants. Posttraumatic epilepsy can arise through a number of mechanisms, which often coexist within a single patient. Penetrating brain injury produces a cicatrix in the cortex and is associated with a risk of posttraumatic epilepsy of approximately 50%, whereas nonpenetrating head injury may produce focal contusions and intracranial hemorrhages, and is associated with a risk of posttraumatic epilepsy of up to 30%. Furthermore, closed head injury often produces diffuse concussive injury, with shearing of axons and selective damage to vulnerable brain regions, such as the hippocampus. The clinical, neurophysiologic, imaging, and neuropathologic features or epileptogenicity differ between these alternate mechanisms. It is likely that better understanding of the subtypes of epilepsy resulting from brain trauma will be required to successfully identify antiepileptogenic therapies. PMID- 19187291 TI - From traumatic brain injury to posttraumatic epilepsy: what animal models tell us about the process and treatment options. AB - A large number of animal models of traumatic brain injury (TBI) are already available for studies on mechanisms and experimental treatments of TBI. Immediate and early seizures have been described in many of these models with focal or mixed type (both gray and white matter damage) injury. Recent long-term video electroencephalography (EEG) monitoring studies have demonstrated that TBI produced by lateral fluid-percussion injury in rats results in the development of late seizures, that is, epilepsy. These animals develop hippocampal alterations that are well described in status epilepticus-induced spontaneous seizure models and human posttraumatic epilepsy (PTE). In addition, these rats have damage ipsilaterally in the cortical injury site and thalamus. Although studies in the trauma field provide a large amount of information about the molecular and cellular alterations corresponding to the immediate and early phases of PTE, chronic studies relevant to the epileptogenesis phase are sparse. Moreover, despite the multiple preclinical pharmacologic and cell therapy trials, there is no information available describing whether these therapeutic approaches aimed at improving posttraumatic recovery would also affect the development of lowered seizure threshold and epilepsy. To make progress, there is an obvious need for information exchange between the trauma and epilepsy fields. In addition, the inclusion of epilepsy as an outcome measure in preclinical trials aiming at improving somatomotor and cognitive recovery after TBI would provide valuable information about possible new avenues for antiepileptogenic interventions and disease modification after TBI. PMID- 19187293 TI - Posttraumatic epilepsy: the challenge of translating discoveries in the laboratory to pathways to a cure. AB - Translating laboratory discoveries into successful therapies for preventing epilepsy is a difficult task, but preventing epilepsy in those who are known to be at high risk needs to be one of our highest priorities. At present, we need to approach this task as a parallel set of research endeavors-one concentrating on laboratory experiments designed to learn how to prevent epilepsy after brain trauma and the other focusing on how to perform the appropriate clinical research in humans to demonstrate that whatever is discovered in the laboratory can be appropriately tested. It is too important to let the second process await conclusion of the first. Initially, we need to create a consortium of groups in trauma centers that are dedicated to antiepileptogenic studies and develop funding sources for long-term studies. We need to experiment with clinical protocols, making the studies as cost-effective as possible, while performing continuous data mining of outcomes and surrogate markers. The limitations of current technology to assist in antiepileptogenesis trials must be acknowledged: There is no currently available method for continuously monitoring electroencephalography (EEG) over prolonged periods, and there are no validated biomarkers for the process of epileptogenesis. As we learn more about the process of epileptogenesis and its underlying mechanisms, it is hoped that we will be able to prevent the development of epilepsy after traumatic brain injury (TBI) and after many other known epileptogenic lesions. PMID- 19187294 TI - Analysis of thromboembolic risk related to high-dose intravenous immunoglobulin treatment: a preliminary clinical study of 10 patients with autoimmune mucocutaneous blistering diseases. AB - BACKGROUND: Intravenous immunoglobulin (IVIg) treatment is a well-known treatment that has been used successfully in a broad spectrum of autoimmune diseases. Currently no data are available in the literature about the role of IVIg in the pathogenesis of thromboembolic events in patients with autoimmune blistering diseases refractory to conventional immunosuppressive treatment. AIM: To determine the relationship between IVIg and thromboembolism in patients with autoimmune blistering diseases and to establish a protocol to deal with the thromboembolic risk. METHODS: In our preliminary clinical study, 10 patients with autoimmune blistering diseases underwent IVIg cycles to a total of 133 cycles in all (total number of infusions in the patient group: 399), at a standard dose of 2 g/kg/infusion accompanied by an accurate and a complete clinical and laboratory screening for thromboembolism. Preventive measures, such as hydration before and after IVIg, and administration of 100 mg of acetyl salicylic acid (aspirin) or 1000 IU of subcutaneous heparin calcium per day for 3 weeks, were introduced to reduce the thromboembolic risk. RESULTS: Throughout the 2 years of IVIg treatment, no patient developed a superficial and/or deep venous or arterial thrombosis, even though some of the patients had underlying thromboembolic risk factors and had tested positive for some congenital and acquired thrombophilia markers. CONCLUSIONS: Our results indicate that thromboembolic events are uncommon, despite the presence of risk factors. However, as these disorders are very rare and the percentage of nonresponder patients is very low, further investigations are needed to better understand whether IVIg alone is able to trigger these fatal events in blistering disorders. PMID- 19187292 TI - Epilepsy following cortical injury: cellular and molecular mechanisms as targets for potential prophylaxis. AB - The sequelae of traumatic brain injury, including posttraumatic epilepsy, represent a major societal problem. Significant resources are required to develop a better understanding of the underlying pathophysiologic mechanisms as targets for potential prophylactic therapies. Posttraumatic epilepsy undoubtedly involves numerous pathogenic factors that develop more or less in parallel. We have highlighted two potential "prime movers": disinhibition and development of new functional excitatory connectivity, which occur in a number of animal models and some forms of epilepsy in humans. Previous experiments have shown that tetrodotoxin (TTX) applied to injured cortex during a critical period early after lesion placement can prevent epileptogenesis in the partial cortical ("undercut") model of posttraumatic epilepsy. Here we show that such treatment markedly attenuates histologic indices of axonal and terminal sprouting and presumably associated aberrant excitatory connectivity. A second finding in the undercut model is a decrease in spontaneous inhibitory events. Current experiments show that this is accompanied by regressive alterations in fast-spiking gamma aminobutyric acid (GABA)ergic interneurons, including shrinkage of dendrites, marked decreases in axonal length, structural changes in inhibitory boutons, and loss of inhibitory synapses on pyramidal cells. Other data support the hypothesis that these anatomic abnormalities may result from loss of trophic support normally provided to interneurons by brain-derived neurotrophic factor (BDNF). Approaches that prevent these two pathophysiologic mechanisms may offer avenues for prophylaxis for posttraumatic epilepsy. However, major issues such as the role of these processes in functional recovery from injury and the timing of the critical period(s) for application of potential therapies in humans need to be resolved. PMID- 19187295 TI - Classifying patients with Behcet's disease for disease severity, using a discriminating analysis method. AB - BACKGROUND: Interleukin (IL)-8 has been shown to correlate with the activity of Behcet's disease (BD). The aim of this study was to develop tools as reliable as IL-8 levels in defining BD severity. METHODS: In total, 32 patients with BD and 16 healthy controls were included in the study. Medical history, physical examination, routine laboratory investigations and measurement of serum IL-8 levels were performed. The patients were classified as groups I and II based on the serum IL-8 levels. The patients were then reassigned to groups based on the predictions through discriminating analysis. RESULTS: In addition to the differences between patient and control groups, the differences between groups I and II, group I and controls, and group II and controls were statistically significant. The discriminating analysis results showed that the patients had been assigned to groups I and II with 100% accuracy. CONCLUSION: Discriminating analysis using clinical and routine laboratory findings indicated that 100% of the patients were accurately assigned to the same groups as their IL-8 levels indicated. This suggests that the classification made according to discriminating analysis using these routine variables is a reliable method in determination of the disease severity. PMID- 19187296 TI - Prevalence of cutaneous bacterial infections and nasal carriage of Staphylococcus aureus in recipients of renal transplants. AB - BACKGROUND: Renal transplant recipients (RTRs) often develop bacterial infections as a result of their long-term immunosuppressive treatment. However, there is no published case-control study of cutaneous bacterial infections in this population, and the prevalence of nasal Staphyloccus aureus carriage and its role in cutaneous bacterial infections in RTRs are not known. AIMS: To determine whether the prevalence of cutaneous bacterial infections and nasal S. aureus carriage are increased in RTRs and to investigate the association between nasal S. aureus carriage and cutaneous staphylococcal infections. METHODS: In total, 66 outpatient RTRs and 67 controls were investigated for the presence of cutaneous bacterial infections. Bacterial cultures were taken from clinically suspicious cutaneous lesions, and three nasal swabs were collected to detect nasal S. aureus colonization. RESULTS: Cutaneous bacterial infection was suspected in 42.4% of RTRs, and in 14.2% of controls. However, of the lesions that could be cultured, microbiologically proven cutaneous bacterial [methicillin-sensitive S. aureus (MSSA)] infections were confirmed in only two RTRs and one control subject. Nasal S. aureus carriage was found in 10.6% of RTRs and 29.9% of controls (P < 0.05). Both RTRs with MSSA infection were nasal carriers, whereas nasal S. aureus carriage was not detected in the only control subject with MSSA infection. All S. aureus isolates were oxacillin-sensitive. CONCLUSION: Screening for nasal S. aureus carriage does not seem to assist in preventing staphylococcal bacterial infections in outpatient RTRs. PMID- 19187297 TI - Antineutrophil cytoplasmic antibody-associated vasculitis with oculomotor nerve palsy. AB - We report a patient with antineutrophil cytoplasmic antibody-associated vasculitis with oculomotor nerve palsy. The patient presented with a high fever, diplopia, blepharoptosis and impairment of ocular movement of the left eye except for lateral gaze. Multiple erythematous and livedoid lesions were observed on the forehead, both cheeks and both legs. Laboratory examination showed positive results for myeloperoxidase antineutrophil cytoplasmic antibodies. Skin biopsy revealed leucocytoclastic vasculitis of the small arteries in the lower dermis. The patient was successfully treated with systemic corticosteroids. PMID- 19187298 TI - Multicentric reticulohistiocytosis with lungs and liver involved. AB - We describe a 56-year-old man with a 2-year history of papulonodules, pruritic and painful on palpation, on the head, trunk, limbs, buttocks and scrotum and a 1.5-year history of rheumatoid arthritis-like joint changes. Biopsies from the nodules on the head and left elbow revealed multinucleated giant cells with eosinophilic 'ground-glass' cytoplasm. Computed tomography revealed that there were scattered nodules in the liver and both lungs. Biopsies taken from nodules in the right lung and liver were consistent with multicentric reticulohistiocytosis. The widely scattered cutaneous papulonodules and the generalized systemic involvement make this patient interesting, and the condition should be differentiated from other diseases in clinicopathological practice. PMID- 19187299 TI - Traumatic lingual ulcer in a child: Riga-Fede disease. AB - Riga-Fede disease RFD is an extremely rare, benign inflammatory disorder characterized by reactive, traumatic ulceration of the oral mucosa especially located on the tongue. It is most commonly associated with natal or neonatal teeth in newborns. Mucosal lesions are often caused by repetitive traumatic damage due to backward and forward motions of the tongue over the lower incisors. Failure to diagnose and treat these lesions properly may result in inadequate food intake, growth retardation and permanent lingual deformity. We report a 15 month-old healthy infant with tongue ulcer diagnosed as RFD based on history and clinical features. PMID- 19187300 TI - Association of Sweet's syndrome and acute sarcoidosis: report of a case and review of the literature. AB - Sweet's syndrome (acute febrile neutrophilic dermatosis, SS) may be considered a reactional dermatosis with numerous associated diseases. We describe a rare association between SS and acute sarcoidosis in a 55-year-old woman. The atypical feature of positive tuberculin reaction is discussed. In this association, diagnosis may be challenging with coexistent skin lesions of both diseases. Acute sarcoidosis should be considered when SS is accompanied with mediastinal lymphadenopathy, uveitis or erythema nodosum. Deep-seated nodular lesions in the context of SS are not always 'deep' SS lesions. PMID- 19187301 TI - Bullous pemphigoid in an infant using complementary medicine. AB - Bullous pemphigoid (BP) is an acquired immunobullous disorder rarely seen in childhood. We report the case of an infant with BP successfully treated with oral corticosteroids. The onset of BP was associated with use of complementary medications and we speculate that these may have been triggering factors. PMID- 19187302 TI - Recurrent bullous lesions associated with familial Mediterranean fever: a case report. AB - Familial Mediterranean fever (FMF) is an inherited, recurrent, inflammatory disease. Of its various cutaneous features, erysipelas-like erythema is the best known and most common skin lesion. We present a new case of FMF with recurrent bullous lesions. A 41-year-old woman was admitted to our clinic with tense bullae, 20 x 20 mm in diameter on the left shin. The patient had a history of fever, abdominal pain, peritonitis attacks and infertility. A lesional skin biopsy revealed subepidermal bullae and neutrophilic infiltration around dermal vessels. Direct immunofluorescence analysis was negative. Over the period of investigation, the lesion regressed spontaneously; 1 month later, a similar lesion appeared on the right wrist. Diagnosis of FMF was made according to the Tel-Hashomer criteria. Recognition of this peculiar skin lesion may lead to an earlier diagnosis of the disease. PMID- 19187303 TI - Severe acneiform eruption exacerbated by dantrolene sodium. PMID- 19187304 TI - Trichoblastoma of the ala nasi: a case report. PMID- 19187305 TI - A case of unexpected facial tattooing following intra-oral injection of Indian ink. PMID- 19187306 TI - Multiple adult myofibromas with congenital smooth muscle hamartoma. PMID- 19187307 TI - A case of ;creeping hair' resembling cutaneous larva migrans. PMID- 19187308 TI - Acquired digital fibrokeratoma associated with ciclosporin treatment. PMID- 19187309 TI - Anetoderma accompanying microscopic panniculitis in a patient with antiphospholipid syndrome and systemic lupus erythematosus. PMID- 19187310 TI - A case of mid-dermal elastolysis after narrowband ultraviolet B phototherapy. PMID- 19187311 TI - All patients with sebaceous gland neoplasms should be screened for Muir-Torre syndrome. PMID- 19187312 TI - Relapsing erysipelas of the buttock due to Streptococcus agalactiae in an immunocompetent woman. PMID- 19187313 TI - Plantar herald patch in pityriasis rosea. PMID- 19187314 TI - Peculiar progressive plaques. PMID- 19187315 TI - A longstanding nail abnormality in a young man. PMID- 19187316 TI - A post-transplant complication. PMID- 19187317 TI - Papular eruption in an elderly woman. PMID- 19187318 TI - Bulbous swelling on the finger pulp. PMID- 19187319 TI - Silver hair in a 3-year-old. PMID- 19187322 TI - New frontiers in allergen standardization. PMID- 19187323 TI - On the origin and specificity of antibodies to neuromuscular blocking (muscle relaxant) drugs: an immunochemical perspective. AB - Following the demonstration 25 years ago that substituted ammonium groups on neuromuscular blocking drugs (NMBDs) are the main allergenic structures recognized by IgE antibodies in the sera of some patients who experience anaphylaxis during anaesthesia, immunoassays for these drugs were quickly applied to supplement skin tests in the diagnostic assessment of suspected adverse reactions to anaesthetic agents. Many subjects who react to an NMBD do so on first exposure and this led to the speculation that the origin of allergic sensitization is an environmental agent(s) or another drug containing an ammonium ion. Direct antibody binding and hapten inhibition studies revealed that morphine, which contains a tertiary amino group, was strongly recognized by IgE in sera from anaphylactic patients and a morphine-solid phase immunoassay was found to be superior to NMBD-based assays for the detection of NMBD-reactive IgE antibodies. Extensive inhibition experiments indicate the likelihood of antibody combining site heterogeneity with recognition at the fine structural level of features additional, and adjacent to, ammonium ions. Further quantitative investigations are needed to identify these neighbouring groups on different NMBDs. Recent work has implicated the morphine analogue pholcodine as the sensitizing agent in Norway where, unlike Sweden, anaphylactic reactions to NMBDs are not uncommon and the medicament is available over-the-counter. This has led to the suggestion that allergenic sensitization to the ammonium group of pholcodine may account for the different incidences of anaphylaxis during anaesthesia in the two countries. This work is subjected to critical review and some alternative speculations on the nature and origin of the sensitizing agent(s) are presented. PMID- 19187325 TI - Relevance of the hygiene hypothesis to early vs. late onset allergic rhinitis. AB - INTRODUCTION: The hygiene hypothesis proposes that reduced exposure to infections in early life increases the risk of developing allergic conditions including allergic rhinitis. We examined the association between markers of the hygiene hypothesis and allergic rhinitis that developed before 7 years of age and allergic rhinitis that developed after 7 years of age. METHODS: The Tasmanian Longitudinal Health Study (TAHS) is a population-based cohort (n=8583) study of respiratory disease. Participants have been followed from 7 to 44 years of age. Information on potential risk factors, allergies and respiratory symptoms was collected longitudinally. Using multi-nomial logistic regression, exposure to siblings, infections, tonsillectomy and farm residence during childhood were examined as risk factors for allergic rhinitis that developed before or after 7 years of age. All analyses were adjusted for gender, maternal and paternal atopy, mother's age at participant's birth, paternal socio-economic status in 1968 and personal socio-economic status in 2004. RESULTS: Greater cumulative exposure to siblings before the age of 2 years was strongly inversely associated with early onset allergic rhinitis (<1 year sib exposure: OR=0.6, 95% CI 0.3-1.0; 1-3 years sib exposure: OR=0.6, 95% CI 0.4-0.9; >3 years sib exposure: OR=0.4, 95% CI 0.3 0.8) less so with later onset allergic rhinitis. The risk of early onset allergic rhinitis decreased with increasing viral infections (OR=0.7, 95% CI 0.5-0.9) during childhood. Having a tonsillectomy before 7 years of age increased the risk of early onset allergic rhinitis (OR=1.7, 95% CI 1.2-2.5). None of these factors was associated with later onset allergic rhinitis. CONCLUSIONS: Exposures relevant to the hygiene hypothesis were important predictors for the development of early onset but less so for later onset allergic rhinitis. The exact mechanisms by which siblings and infections protect against allergic rhinitis are unclear. The stronger findings for earlier onset allergic rhinitis suggest that family structure and infections have most impact on disease risk in early life. Further research should focus on early onset allergic rhinitis when exploring causal explanations for any sibling effect. PMID- 19187324 TI - Inflammatory and oxidative stress biomarkers in allergic rhinitis: the effect of smoking. AB - BACKGROUND: Accumulating evidence confirms the presence of pan-airway inflammation in allergic rhinitis patients. Smoking is known to affect the asthmatic airway inflammation. However, no study has evaluated the impact of smoking on airway inflammation of allergic rhinitis patients. OBJECTIVE: The aim of the present study was to evaluate the impact of smoking on inflammatory and oxidative stress biomarkers in patients with seasonal allergic rhinitis, using non-invasive methods for sample collection. METHODS: Forty patients with seasonal allergic rhinitis (20 smokers and 20 non-smokers) and 30 healthy subjects (15 smokers and 15 non-smokers) were recruited for the study during pollen season. All subjects were submitted to measurement of the fraction of exhaled NO (FeNO), exhaled breath condensate (EBC) collection, nasal lavage collection, pre- and post- bronchodilation spirometry and metacholine bronchial challenge testing. pH, leukotriene B(4) (LTB(4)) and 8-isoprostane were determined in EBC and nasal lavage samples. RESULTS: Patients with allergic rhinitis presented higher LTB(4) and 8-isoprostane levels in nasal lavage (P<0.0001 for both comparisons), with no significant differences between smokers and non-smokers. Patients with allergic rhinitis also presented higher LTB(4) levels and lower pH in EBC (P<0.001 and P=0.004, respectively), with prominent differences between smokers and non smokers (P<0.0001 and P=0.003, for LTB(4) and pH, respectively). A significant correlation between nasal lavage and EBC LTB(4) values was observed (r(s)=0.313, P=0.048). CONCLUSIONS: Patients with allergic rhinitis present increased LTB(4) and 8-isoprostane in their nasal cavity, however, with no significant differences between smokers and non-smokers. In contrast, smokers with allergic rhinitis present higher LTB(4) levels and lower pH in EBC, suggesting that these patients may be more susceptible to the deleterious effects of smoking, compared with non smokers. PMID- 19187326 TI - Relationship between oxidative stress-related biomarkers and antioxidant status with asthma and atopy in young adults: a population-based study. AB - BACKGROUND AND AIM: Enhanced oxidative stress has been described in adults who suffer from symptoms of asthma and poor lung function. This study assessed the relation between markers of oxidative stress and antioxidant status and lung function, symptoms of asthma, atopy and bronchial hyperresponsiveness (BHR) in young adults. METHODS: A sub-sample of 589 individuals aged 22-28 years, selected from a total of 1232 included in a survey assessing early and current risk factors for chronic diseases, participated in the study. Participants were from an agricultural area of Chile, responded to a Spanish version of the European Community Respiratory Health Survey questionnaire, were skin tested to eight allergens, and challenged with methacholine to assess BHR. Five hundred and eighty-five individuals had measures of plasma biomarkers ferric reducing ability of plasma, uric acid, protein carbonyls and 564 had 8-iso-prostaglandin F(2alpha) (8-iso-PGF(2alpha)) assessed. RESULTS: All participants had detectable plasma 8 iso-PGF(2alpha) and carbonyl levels. There was no indication for an association between markers of antioxidant status or oxidative stress with any of the outcomes studied. CONCLUSION: The levels of oxidative stress-related biomarkers and antioxidant status in plasma may not be related to asthma in the general population in the absence of more severe symptoms or exacerbations. PMID- 19187327 TI - Effect of levocetirizine on nasal provocation testing with adenosine monophosphate compared with allergen challenge in allergic rhinitis. AB - BACKGROUND: End-organ hyperreactivity is an important feature of the allergic airway. There are no data directly comparing the responsiveness to treatment of different nasal provocation tests (NPT). OBJECTIVE: We compared the effect of levocetirizine on nasal adenosine 5'-monophosphate (AMP) with specific allergen challenge in patients with intermittent and persistent allergic rhinitis (AR). METHODS: Patients with AR were randomized in double-blind cross-over fashion to receive single doses of levocetirizine 5 mg or identical placebo, with nasal challenge performed 12 h after dosing. Sixteen participants completed per protocol. Nasal AMP or allergen challenge was conducted on separate days with 1- and 2-week washout periods in between, respectively. Measurements of peak nasal inspiratory flow (PNIF) were made over 60 min after each challenge. The primary end-point was the provocative concentration of AMP or allergen causing a 20% drop in the PNIF (PC(20)). RESULTS: The time-profile for PNIF recovery [area under the 60 min time-response curve as % PNIF change (min)] were significantly attenuated for AMP challenge, as mean difference [95% confidence interval (CI)]: 11.57 (3.87, 19.25), P=0.005 and for allergen challenge: 17.82 (0.11, 35.53), P=0.04. A highly significant correlation was shown between methods for the area under the curve: (R=0.86, P<0.001). A statistically significant correlation was also seen for the PC(20): (R=0.94, P<0.001). PC(20) improvement amounted to a 1.26 (95% CI 0.16, 2.35) and 0.16 (95% CI -0.41, 0.73) doubling-dilution shifts for allergen and AMP challenges, respectively. Bland-Altman plots confirmed good agreement between methods. CONCLUSION: A high correlation and statistical agreement has been demonstrated between AMP and allergen challenge for all outcome measures. In particular, the recovery profile after NPT is a sensitive and discriminatory measure of anti-allergic treatment. PMID- 19187329 TI - New insights into preschool wheeze. PMID- 19187330 TI - Provoked models of asthma: what have we learnt? AB - Asthma is a chronic inflammatory disease of the airways characterized by physiological abnormalities of variable airflow obstruction and airway hyperresponsiveness (AHR) to a wide variety of physical and inhaled chemical stimuli and the presence of symptoms. AHR is measured by challenging the airways with a variety of agonists and naturally occurring stimuli, which results in constriction of the airway smooth muscle, leading to airway narrowing and airflow limitation. There are two distinct mechanisms by which the airways can narrow to a constrictor stimulus and these are defined by the pathways they take to induce AHR. Direct stimuli are pharmacological agents administered exogenously (such as histamine or methacholine) that act 'directly' on specific receptors on the bronchial smooth muscle to cause constriction. The other mechanism by which the airway can narrow is via the inhalation of indirect stimuli, which include natural stimuli, such as allergen or exercise, and pharmacological agents such as adenosine monophosphate and hyper-osmotic agents (e.g. hypertonic saline or dry powder mannitol). These stimuli induce airway narrowing 'indirectly' by causing the endogenous release of mediators of bronchoconstriction from airway inflammatory cells. Provoked models of asthma have been extremely valuable in understanding the pathobiology of asthma, in aiding diagnosis, in helping to clarify the mechanisms of actions of effective drugs and in the development of new entities to treat asthma. Some provoked models are valuable clinically, particularly those that measure direct AHR, while others, particularly allergen challenge, have been used in animal models and in humans to study the mechanisms of allergen-induced airway inflammation and the associated physiological changes, as well in the development of new drugs for asthma. An emerging role for measurements of AHR is in the evaluation of the optimal treatment for patients with asthma. PMID- 19187332 TI - Alpha-T-catenin (CTNNA3) gene was identified as a risk variant for toluene diisocyanate-induced asthma by genome-wide association analysis. AB - BACKGROUND: Toluene diisocyanate (TDI) is the most important cause of occupational asthma, but the genetic mechanism of TDI-induced asthma is still unknown. OBJECTIVE: The objective of the study was to identify susceptibility alleles associated with the TDI-induced asthma phenotype. METHODS: We conducted a genome-wide association study in 84 patients with TDI-induced asthma and 263 unexposed healthy normal controls using Affymetrix 500K SNPchip. We also investigated the relationships between genetic polymorphisms and transcript levels in Epstein-Barr virus-transformed lymphoblastoid cell lines from patients with TDI-induced asthma enrolled in this study. RESULTS: Genetic polymorphisms of CTNNA3 (catenin alpha 3, alpha-T catenin) were significantly associated with the TDI-induced asthma phenotype (5.84 x 10(-6) for rs10762058, 1.41 x 10(-5) for rs7088181, 2.03 x 10(-5) for rs4378283). Carriers with the minor haplotype, HT2 [GG], of two genetic polymorphisms (rs10762058 and rs7088181) showed significantly lower PC(20) methacholine level (P=0.041) and lower mRNA expression of CTNNA3 than non-carriers (P=0.040). A genetic polymorphism in the 3' downstream region of CTNNA3 (rs1786929), as identified by DNA direct sequencing, was significantly associated with the TDI-induced asthma phenotype (P=0.015 in recessive analysis model) and the prevalence of serum-specific IgG to cytokeratin 19 (P=0.031). CONCLUSION: These findings suggested that multiple genetic polymorphisms of CTNNA3 may be determinants of susceptibility to TDI-induced asthma. PMID- 19187331 TI - Acute exacerbations of asthma: epidemiology, biology and the exacerbation-prone phenotype. AB - Asthma is a highly prevalent chronic respiratory disease affecting 300 million people world-wide. A significant fraction of the cost and morbidity of asthma derives from acute care for asthma exacerbations. In the United States alone, there are approximately 15 million outpatient visits, 2 million emergency room visits, and 500,000 hospitalizations each year for management of acute asthma. Common respiratory viruses, especially rhinoviruses, cause the majority of exacerbations in children and adults. Infection of airway epithelial cells with rhinovirus causes the release of pro-inflammatory cytokines and chemokines, as well as recruitment of inflammatory cells, particularly neutrophils, lymphocytes, and eosinophils. The host response to viral infection is likely to influence susceptibility to asthma exacerbation. Having had at least one exacerbation is an important risk factor for recurrent exacerbations suggesting an 'exacerbation prone' subset of asthmatics. Factors underlying the 'exacerbation-prone' phenotype are incompletely understood but include extrinsic factors: cigarette smoking, medication non-compliance, psychosocial factors, and co-morbidities such as gastroesophageal reflux disease, rhinosinusitis, obesity, and intolerance to non-steroidal anti-inflammatory medications; as well as intrinsic factors such as deficient epithelial cell production of the anti-viral type I interferons (IFN alpha and IFN-beta). A better understanding of the biologic mechanisms of host susceptibility to recurrent exacerbations will be important for developing more effective preventions and treatments aimed at reducing the significant cost and morbidity associated with this important global health problem. PMID- 19187333 TI - Expression profiling of genes related to asthma exacerbations. AB - BACKGROUND: Asthma is a chronic airway inflammatory disease; however, the molecular mechanisms that underlie asthma exacerbation are only partially understood. OBJECTIVE: To identify gene expression signatures that reflect the acute exacerbation of asthma, we examined the differential expression of genes during asthma exacerbation and stable condition by using microarray analysis. METHODS: The subjects were mite-sensitive asthmatic children and non-asthmatic control children. The children were divided into four groups (AE: asthma exacerbation, n=12; SA: stable asthma, n=11; IC: infected control, n=6; and NC: non-infected control, n=5). Total RNA was extracted from peripheral blood mononuclear cells and subjected to microarray analysis with Illumina Human Ref8 BeadChip arrays. Welch's t-test was performed to identify genes whose expression was altered during asthma exacerbation. Quantitative real-time RT-PCR was performed on samples collected from 43 asthmatic children and 11 control children to verify the microarray results. RESULTS: The expression of 137/16 genes was significantly up/down-regulated during asthma exacerbation assessed by microarray analysis. Of the genes, 62 were also differentially expressed during upper respiratory infection. Many of the asthma exacerbation related genes were involved in defence responses and responses to external stimuli, but these associations disappeared after excluding the infection-related genes. Quantitative real-time RT-PCR confirmed that the genes related (S100A8 and GAS6) and unrelated to infections (CD200 and RBP7) were differentially expressed during asthma exacerbation (P<0.01). CONCLUSIONS: Previously unidentified immune responses during asthma exacerbation may provide further clarification of the molecular mechanisms underlying asthma. PMID- 19187335 TI - Respiratory dysfunction in chronic neck pain patients. A pilot study. AB - The aim of this pilot study was to add weight to a hypothesis according to which patients presenting with chronic neck pain could have a predisposition towards respiratory dysfunction. Twelve patients with chronic neck pain and 12 matched controls participated in this study. Spirometric values, maximal static pressures, forward head posture and functional tests were examined in all subjects. According to the results, chronic neck patients presented with a statistically significant decreased maximal voluntary ventilation (P = 0.042) and respiratory muscle strength (Pimax and Pemax), (P = 0.001 and P = 0.002, respectively). Furthermore, the current study demonstrated a strong association between an increased forward head posture and decreased respiratory muscle strength in neck pateits. The connection of neck pain and respiratory function could be an important consideration in relation to patient assessment, rehabilitation and consumption of pharmacological agents. PMID- 19187334 TI - Food allergy is associated with an increased risk of asthma. AB - BACKGROUND: The atopic march is well documented, but the interrelationship of food allergy (FA) and asthma is not well understood. OBJECTIVE: The aim of this study was to examine the strength of the association and temporal relationships between FA and asthma. METHODS: This analysis included 271 children >or=6 years (older group) and 296 children <6 years (younger group) from a family-based FA cohort in Chicago, IL. Asthma was determined by parental report of physician diagnosis. FA status was determined based on the type and timing of clinical symptoms after ingestion of a specific food, and results of prick skin test (Multi-Test II) and allergen-specific IgE (Phadia ImmunoCAP). Analyses were carried out using logistic regression accounting for important covariates and auto-correlations among siblings. Kaplan-Meier curves were used to compare the time to onset of asthma with the FA status. RESULTS: Symptomatic FA was associated with asthma in both older [odds ratio (OR)=4.9, 95% confidence interval (CI): 2.5-9.5] and younger children (OR=5.3, 95% CI: 1.7-16.2). The association was stronger among children with multiple or severe food allergies, especially in older children. Children with FA developed asthma earlier and at higher prevalence than children without FA (Cox proportional hazard ratio=3.7, 95% CI: 2.2-6.3 for children >or=6 years, and hazard ratio=3.3, 95% CI: 1.1-10 for children <6 years of age). No associations were seen between asymptomatic food sensitization and asthma. CONCLUSIONS: Independent of markers of atopy such as aeroallergen sensitization and family history of asthma, there was a significant association between FA and asthma. This association was even stronger in subjects with multiple food allergies or severe FA. PMID- 19187336 TI - Whites and African-Americans in headache specialty clinics respond equally well to treatment. AB - This study sought to determine if Whites and African-Americans respond similarly to headache treatment administered in 'real-world' headache specialty treatment clinics. Using a naturalistic, longitudinal design, 284 patients receiving treatment for headache disorders completed 30-day daily diaries that assessed headache frequency and severity at pretreatment and 6-month follow-up and also provided data on their headache disability and quality of life at pretreatment and 1-, 2- and 6-month follow-up. Controlling for socioeconomic status and psychiatric comorbidity, hierarchical linear models found that African-Americans and Whites reported significant reductions in headache frequency and disability and improvements in life quality over the 6-month treatment period. African Americans, unlike Whites, also reported significant decreases in headache severity. Nevertheless, Africans-Americans had significantly more frequent and disabling headaches and lower quality of life after treatment relative to Whites. Although Whites and African Americans responded favourably to headache treatments, more efficacious treatments are needed given the elevated level of headache frequency that remained in both racial groups following treatment. PMID- 19187337 TI - A nationwide population-based study of migraine in Brazil. AB - The aim of this study was to estimate the 1-year prevalence of migraine and the degree of the association of migraine with some sociodemographic characteristics of a representative sample of the adult population of Brazil. This was a cross sectional, population-based study. Telephone interviews were conducted on 3848 people, aged 18-79 years, randomly selected from the 27 States of Brazil. The estimated 1-year gender- and age-adjusted prevalence of migraine was 15.2%. Migraine was 2.2 times more prevalent in women, 1.5 times more in subjects with > 11 years of education, 1.59 times more in subjects with income of < 5 Brazilian Minimum Wages per month, and 1.43 times more in those who do not do any physical exercise. The overall prevalence of migraine in Brazil is 15.2%. Migraine is significantly more prevalent in women, subjects with higher education, with lower income, and those who do not exercise regularly, independently of their body mass index. PMID- 19187338 TI - Morning headache in sleep apnoea: clinical and polysomnographic evaluation and response to nasal continuous positive airway pressure. AB - Morning headache is accepted as part of clinical findings of obstructive sleep apnoea syndrome (OSAS). The prevalence of morning headache is at variable levels from 18% to 74% in patients with OSAS. However, there is controversy over the association of morning headache and OSAS. We studied morning headache prevalence and characteristics in 101 controls with apnoea-hypnoea index (AHI) < 5 and 462 OSAS patients with AHI > or = 5. Morning headache was reported by only nine (8.9%) subjects in a control group compared with 156 (33.6%) of OSAS patients (P < 0.01). Morning headache prevalence was significantly higher in severe and moderate OSAS groups. AHI was significantly higher in OSAS patients with morning headache compared with patients without morning headaches. Oxygen saturation nadir during rapid eye movement and non-rapid eye movement sleep as well as mean oxygen saturation value during total sleep time were also found to be significantly lower in morning headache group. However, none of the sleep parameters was found to be determinants of morning headache. Morning headache was more frequently reported by patients of female gender and with primary headache history. Morning headache was totally resolved in 90% of patients treated with nasal continuous positive airway pressure. The history of OSAS should be considered in the differential diagnosis of morning headache. PMID- 19187339 TI - Visual attention in children with migraine: a controlled comparative study. AB - The aim of this study was to evaluate the visual attention of children with migraine and compare it with a control group. Thirty migrainous children and 30 controls without headache were subjected to a visual attention assessment with Trail Making Tests (TMT) A/B, Letter Cancellation Test, and the Brazilian computerized test Visual Attention Test, third edition. The migraine group was evaluated after 2 days without headache. The migraine group had an inferior performance compared with the control group on TMT A (P = 0.03) and B (P = 0.001), and more errors on tasks 1 (P = 0.032) and 2 (P = 0.015) of the Visual Attention Test, presenting difficulty with selective and alternate attention. Attention is a neurological function that depends on structures such as the brainstem, cerebral cortex and the limbic system and on neurotransmitters such as dopamine and noradrenaline. The neurochemical aspects involved in the physiopathology of migraine and attention mechanisms probably predispose these children to visual attention deficits. PMID- 19187340 TI - Prostaglandin E2(PGE2) induces headache in healthy subjects. AB - The role of prostanoids in nociception is well established. The headache eliciting effects of prostaglandin E(2) (PGE(2)) and its possible mechanisms have previously not been systematically studied in man. We hypothesized that infusion of PGE(2) might induce headache and vasodilation of cranial vessels. PGE(2) (0.40 microg kg(-1) min(-1)) or saline was infused for 25 min into 11 healthy subjects in a cross-over, double-blind study. Headache intensity was scored on a verbal rating scale from 0 to 10. In addition, we recorded mean flow in the middle cerebral artery (V(MCA)) by transcranial Doppler and diameter of the superficial temporal artery (STA) by high-resolution ultrasonography. All 11 subjects reported headache on the PGE(2) day and no subjects reported headache on the placebo day (P = 0.001). During the immediate phase (0-30 min) (P = 0.005) and the postinfusion phase (30-90 min) (P = 0.005), the area under the curve for headache score was significantly larger on the PGE(2) day compared with the placebo day. PGE(2) caused dilatation of the STA (23.5%; 95% CI 14.0, 37.8) and the MCA (8.3%; 95% CI 4.0, 12.6). We suggest that PGE(2) induces headache by activation and sensitization of cranial perivascular sensory afferents. PMID- 19187345 TI - Unusual manifestations of tegumentary leishmaniasis in AIDS patients from the New World. AB - BACKGROUND: Comorbidity from tegumentary leishmaniasis and AIDS is poorly characterized. OBJECTIVES: To describe a series of patients coinfected with Leishmania and human immunodeficiency virus (HIV). METHODS: Clinical records from patients were analysed by demographic data, clinical manifestations, diagnoses, treatments and outcomes. RESULTS: Fifteen cases of AIDS/tegumentary leishmaniasis were found. The diagnosis of leishmaniasis was confirmed by the detection of Leishmania amastigotes or antigens from the cutaneous or mucosal lesions. The mean CD4+ T-cell count was 84 cells mm(-3) (range 8-258) and all patients were classified as having AIDS according to the Centers for Disease Control and Prevention. A wide range of manifestations was found, varying from a single ulcer to multiple and polymorphic lesions. Mucosal lesions were present in 80% and cutaneous lesions in 73% of patients (53% with mucocutaneous form), disseminated lesions in 60% and genital lesions in 27% of patients. All patients received anti Leishmania therapy and 53% showed relapses. Sixty-seven per cent received highly active antiretroviral therapy but showed no difference in outcomes and relapses compared with those not using medication. Forty per cent died during the study period. In these patients, the anti-Leishmania antibody and Montenegro skin test were useful in the diagnosis of leishmaniasis, probably because leishmaniasis preceded immunosuppression due to HIV infection. CONCLUSIONS: Clinical manifestations of tegumentary leishmaniasis in HIV-infected patients are diverse. Our data emphasize possible unusual manifestations of this disease in HIV infected patients, particularly in severely immunosuppressed cases (< 200 CD4+ cells mm(-3)). PMID- 19187348 TI - The risk of prostate cancer amongst South Asian men in southern England: the PROCESS cohort study. PMID- 19187344 TI - On the role of the corticotropin-releasing hormone signalling system in the aetiology of inflammatory skin disorders. PMID- 19187349 TI - Solifenacin in the treatment of urgency and other symptoms of overactive bladder: results from a randomized, double-blind, placebo-controlled, rising-dose trial. PMID- 19187341 TI - Calcium-activated potassium channels and endothelial dysfunction: therapeutic options? AB - The three subtypes of calcium-activated potassium channels (K(Ca)) of large, intermediate and small conductance (BK(Ca), IK(Ca) and SK(Ca)) are present in the vascular wall. In healthy arteries, BK(Ca) channels are preferentially expressed in vascular smooth muscle cells, while IK(Ca) and SK(Ca) are preferentially located in endothelial cells. The activation of endothelial IK(Ca) and SK(Ca) contributes to nitric oxide (NO) generation and is required to elicit endothelium dependent hyperpolarizations. In the latter responses, the hyperpolarization of the smooth muscle cells is evoked either via electrical coupling through myo endothelial gap junctions or by potassium ions, which by accumulating in the intercellular space activate the inwardly rectifying potassium channel Kir2.1 and/or the Na(+)/K(+)-ATPase. Additionally, endothelium-derived factors such as cytochrome P450-derived epoxyeicosatrienoic acids and under some circumstances NO, prostacyclin, lipoxygenase products and hydrogen peroxide (H(2)O(2)) hyperpolarize and relax the underlying smooth muscle cells by activating BK(Ca). In contrast, cytochrome P450-derived 20-hydroxyeicosatetraenoic acid and various endothelium-derived contracting factors inhibit BK(Ca). Aging and cardiovascular diseases are associated with endothelial dysfunctions that can involve a decrease in NO bioavailability, alterations of EDHF-mediated responses and/or enhanced production of endothelium-derived contracting factors. Because potassium channels are involved in these endothelium-dependent responses, activation of endothelial and/or smooth muscle K(Ca) could prevent the occurrence of endothelial dysfunction. Therefore, direct activators of these potassium channels or compounds that regulate their activity or their expression may be of some therapeutic interest. Conversely, blockers of IK(Ca) may prevent restenosis and that of BK(Ca) channels sepsis-dependent hypotension. PMID- 19187350 TI - Are the National Institute for Health and Clinical Excellence guidelines that promulgate active surveillance for low-risk prostate cancer justified by the available evidence? PMID- 19187352 TI - Anterior retraction of the prostate during robotic-assisted laparoscopic radical prostatectomy using the closure device. PMID- 19187353 TI - A conversation between Darracott Vaughan and Marston Linehan. PMID- 19187354 TI - Evidence-based medicine training in residency: a survey of urology programme directors. AB - OBJECTIVE: To investigate whether and in what format urology residents receive formal training in evidence-based clinical practice. METHODS: In 2007 we conducted a survey of the Accreditation Council for Graduate Medical Education (ACGME)-accredited urology residency programmes in the USA. The survey instrument was sent to all programmes, using postal mailing, e-mail and a web-based interface. Programme director and coordinator names and basic demographic information such as programme size and length were retrieved from the institutions' websites and the ACGME database. RESULTS: Of 117 ACGME-accredited urology programmes, 108 responded (92.3%). Most respondents either agreed or strongly agreed with statements that formal evidence-based medicine (EBM) training was valuable to urology residents (97, 89.8%) and should be part of all residency programme curricula (91, 84.3%). Eighty-four programmes (77.8%) indicated that their curricula included formal educational activities related to EBM. These programmes provided a median (interquartile range) of 10 (4.8-25.0) h of EBM teaching per academic year. Most programmes (65.9%) provided fewer hours of EBM training than programme directors perceived as ideal. Asked what would make it easier to integrate EBM into the programme, respondents identified urology-specific educational materials (33.3%), a formal curriculum (24.4%) and faculty development (16.3%) as central needs. CONCLUSION: In this survey we found that most programme directors recognize formal EBM teaching as an important aspect of urology residency training. However, most programmes devote less time to EBM training than they would like, and urology-specific resources have been lacking. These findings support increased efforts to provide urology residents with opportunities to learn EBM-related knowledge and skills in a variety of educational formats. PMID- 19187356 TI - An extra-special Valentine's Day. PMID- 19187357 TI - Association of lower urinary tract symptoms and chronic ischaemia of the lower urinary tract in elderly women and men: assessment using colour Doppler ultrasonography. PMID- 19187358 TI - Infections after prostate biopsy and antibiotic resistance. PMID- 19187360 TI - Laparoscopic transvesical diverticulectomy. PMID- 19187365 TI - Patient-delivered partner therapy for chlamydia: a realistic public health measure in the UK. PMID- 19187366 TI - Systematic analysis of research underfunding in maternal and perinatal health. AB - BACKGROUND: Little published evidence supports the widely held contention that research in pregnancy is underfunded compared with other disease areas. OBJECTIVES: To assess absolute and relative government and charitable funding for maternal and perinatal research in the UK and internationally. SEARCH STRATEGY, SELECTION CRITERIA, DATA COLLECTION, AND ANALYSIS: Major research funding bodies and alliances were identified from an Internet search and discussions with opinion leaders/senior investigators. Websites and annual reports were reviewed for details of strategy, research spend, grants awarded, and allocation to maternal and/or perinatal disease using generic and disease-specific search terms. MAIN RESULTS: Within the imprecision in the data sets, < or =1% of health research spend in the UK was on maternal/perinatal health. Other countries fared better with 1-4% investment, although nonexclusive categorisation may render this an overestimate. In low-resource settings, government funders focused on infectious disease but not maternal and perinatal health despite high relative disease burden, while global philanthropy concentrated on service provision rather than research. Although research expenditure has been deemed as appropriate for 'reproductive health' disease burden in the UK, there are no data on the equity of maternal/perinatal research spend against disease burden, which globally may justify a manyfold increase. AUTHOR'S CONCLUSIONS: This systematic review of research expenditure and priorities from national and international funding bodies suggests relative underinvestment in maternal/perinatal health. Contributing factors include the low political priority given to women's health, the challenging nature of clinical research in pregnancy, and research capacity dearth as a consequence of chronic underinvestment. PMID- 19187368 TI - 'Blood On The Tracks' from corpora lutea to endometriomas. AB - OBJECTIVE: To detect a direct transition from a haemorrhagic corpus luteum to an endometriotic cyst by serial transvaginal ultrasonographic scans. DESIGN: Prospective observational study. SETTING: An academic tertiary care and referral centre for women with endometriosis. POPULATION: One hundred and nine women younger than 40 years, with regular menstrual cycles, undergoing first-line surgery for endometriomas, and not wanting postoperative oral contraception. METHODS: Three-monthly transvaginal ultrasonography during the luteal phase for 2 years after surgery. MAIN OUTCOME MEASURE: Sonographic identification of progression from a haemorrhagic corpus luteum to a recurrent endometriotic cyst. RESULTS: A haemorrhagic corpus luteum was identified in 13 women. Serial ultrasonographic scans demonstrated transition to an endometriotic cyst in 11 (85%) instances and resorption in two. A unilateral endometriotic cyst without previous detection of a cystic corpus luteum was observed in 14 women. CONCLUSIONS: Bleeding from a corpus luteum appears to be a critical event in the development of endometriomas. PMID- 19187367 TI - Preferred strategies of men and women for managing chlamydial infection. AB - OBJECTIVES: To determine men and women's preferred strategies for managing chlamydial infection: partner notification (patient referral), postal testing kit (PTK) or patient-delivered partner medication (PDPM). DESIGN: Interviewer conducted questionnaires (women) and anonymous, self-administered questionnaires (men). POPULATION: Women infected with chlamydia who were participating in a randomised study assigning partners to patient referral, PTK or PDPM. Men attending genitourinary medicine, family planning and fracture clinics in Edinburgh. METHODS: Men and women were asked their preferred strategy for testing/treating sexual partners (patient referral, PTK or PDPM) if they or their partner had a positive chlamydia test. Women were also asked the reasons for their choice and whether partners were satisfied with the intervention received. MAIN OUTCOME MEASURES: Reported preferences of men and women for testing/treating partners. RESULTS: Response rates were 97 and 81% for the women's questionnaires at study entry and 6 months, respectively, and 81% for the men's questionnaires. Of 174 women responding, 67% preferred PDPM for partners and 57% would prefer PDPM for themselves. The main reasons were that PDPM allows simpler, more convenient and faster treatment. Women reported that 65% of partners were satisfied with whichever intervention they received. Of 293 men responding, 70% would choose patient referral for partners and 53% would prefer patient referral for themselves. Men previously tested for chlamydia were significantly more likely to choose PDPM (n = 22) than those never tested (n = 7); P < 0.001. Only 3% of women and 9% of men preferred PTKs for partners. CONCLUSION: The results suggest that women prefer PDPM and men, at least hypothetically, prefer patient referral. PTK appears unpopular with both sexes. PMID- 19187369 TI - The prediction of progression-free and overall survival in women with an advanced stage of epithelial ovarian carcinoma. AB - OBJECTIVE: Prognosis in women with ovarian cancer mainly depends on International Federation of Gynecology and Obstetrics stage and the ability to perform optimal cytoreductive surgery. Since ovarian cancer has a heterogeneous presentation and clinical course, predicting progression-free survival (PFS) and overall survival (OS) in the individual patient is difficult. The objective of this study was to determine predictors of PFS and OS in women with advanced stage epithelial ovarian cancer (EOC) after primary cytoreductive surgery and first-line platinum based chemotherapy. DESIGN: Retrospective observational study. SETTING: Two teaching hospitals and one university hospital in the south-western part of the Netherlands. POPULATION: Women with advanced stage EOC. METHODS: All women who underwent primary cytoreductive surgery for advanced stage EOC followed by first line platinum-based chemotherapy between January 1998 and October 2004 were identified. To investigate independent predictors of PFS and OS, a Cox' proportional hazard model was used. Nomograms were generated with the identified predictive parameters. MAIN OUTCOME MEASURES: The primary outcome measure was OS and the secondary outcome measures were response and PFS. RESULTS: A total of 118 women entered the study protocol. Median PFS and OS were 15 and 44 months, respectively. Preoperative platelet count (P = 0.007), and residual disease <1 cm (P = 0.004) predicted PFS with a optimism corrected c-statistic of 0.63. Predictive parameters for OS were preoperative haemoglobin serum concentration (P = 0.012), preoperative platelet counts (P = 0.031) and residual disease <1 cm (P = 0.028) with a optimism corrected c-statistic of 0.67. CONCLUSION: PFS could be predicted by postoperative residual disease and preoperative platelet counts, whereas residual disease, preoperative platelet counts and preoperative haemoglobin serum concentration were predictive for OS. The proposed nomograms need to be externally validated. PMID- 19187370 TI - Two mifepristone doses and two intervals of misoprostol administration for termination of early pregnancy: a randomised factorial controlled equivalence trial. AB - OBJECTIVE: To compare the efficacy of 100 mg and 200 mg of mifepristone and 24- and 48-hour intervals to administration of 800 microg vaginal misoprostol for termination of early pregnancy. DESIGN: Placebo-controlled, randomized, equivalence trial, stratified by centre. SETTING: 13 departments of obstetrics and gynecology in nine countries. POPULATION: 2,181 women with 63 days or less gestation requesting medical abortion. METHODS: Two-sided 95% CI for the risk differences of failure to complete abortion were calculated and compared with 5% equivalence margin between two doses of mifepristone and two intervals to misoprostol administration. Proportions of women with adverse effects were compared between the regimens using standard testes for proportions. OUTCOME MEASURES: Rates of complete abortion without surgical intervention and adverse effects associated with the regimens. RESULTS: Efficacy outcome was analysed for 2,126 women (97.5%) excluding 55 lost to follow up. Both mifepristone doses were found to be similar in efficacy. The rate of complete abortion was 92.0% for women assigned 100 mg of mifepristone and 93.2% for women assigned 200 mg of mifepristone (difference 1.2%, 95% CI: -1.0 to 3.5). Equivalence was also evident for the two intervals of administration: the rate of complete abortion was 93.5% for 24-hour interval and 91.7% for the 48-hour interval (difference -1.8%, 95% CI: -4.0 to 0.5). Interaction between doses and interval to misoprostol administration was not significant (P = 0.92). Adverse effects related to treatments did not differ between the groups. CONCLUSIONS: Both the 100 and 200 mg doses of mifepristone and the 24- and 48-hour intervals have a similar efficacy to achieve complete abortion in early pregnancy when mifepristone is followed by 800 micrograms of vaginally administered misoprostol. PMID- 19187372 TI - Comparison of maternal mortality between migrating population and permanent residents in Shanghai, China, 1996-2005. AB - OBJECTIVE: Maternal mortality ratio (MMR) in Shanghai residents has been declining in the past two decades and has reached levels comparable to developed countries. The MMR in migrating population in Shanghai remains high, however. The objectives of this study were to compare the trends of MMR between residents and migrating population in Shanghai from 1996 to 2005 and to explore the reasons for the dramatic differences in MMR between the two groups living in the same city. DESIGN: Retrospective cohort study. SETTING: Shanghai, China. POPULATION: A total of 902,807 pregnancies with live births in Shanghai in the period of 1996-2005. METHODS: We first compared the overall MMR between migrating population and permanent residents in Shanghai and examined temporal trends of MMR in the two subpopulations. We then compared the causes and maternal characteristics of maternal deaths between the two subpopulations. MAIN OUTCOME MEASURES: Maternal mortality and cause of death. RESULTS: A total of 902,807 live births and 243 maternal deaths were recorded in Shanghai in the period of 1996 to 2005, with an average MMR of 26.66 per 100,000 live births. The MMR in Shanghai residents declined dramatically from 22.47 per 100,000 in 1996 to 1.64 per 100,000 live births in 2005 (P < 0.01), while the MMR in migrating population was reduced only moderately from 54.68 per 100,000 live births to 48.46 per 100,000 (P > 0.05). The main causes of maternal deaths in migrating population were postpartum haemorrhage (39.9%), pregnancy-induced hypertension (9.8%), and puerperal infection (9.3%), whereas the main causes of maternal death of Shanghai residents were chronic heart and liver diseases (20.0%), postpartum haemorrhage (12.9%), and amniotic fluid embolism (12.9%). Among the maternal death cases in migrating women, 60% had elementary education or less, 22% were unemployed, 65% had no prenatal visit, 44% gave a birth at home, and 12% of the deaths occurred at home. CONCLUSION: Lack of access to quality maternity care, especially for the effective management of postpartum haemorrhage, is the main reason for the high MMR in migrating population in Shanghai. PMID- 19187371 TI - The effect of maternal alcohol consumption on fetal growth and preterm birth. AB - OBJECTIVE: To investigate the relationship between prenatal alcohol exposure and fetal growth and preterm birth and to estimate the effect of dose and timing of alcohol exposure in pregnancy. DESIGN: A population-based cohort study linked to birth information on the Western Australian Midwives Notification System. SETTING: Western Australia. POPULATION: A 10% random sample of births restricted to nonindigenous women who had delivered a singleton infant (n= 4719) in 1995 1997. METHODS: The impact of alcohol consumption in pregnancy on fetal growth (small-for-gestational-age [SGA] and large-for-gestational-age infants [LGA]) and preterm birth (<37 weeks of gestation) was assessed using multivariate logistic regression analysis and adjusting for confounding factors. MAIN OUTCOME MEASURES: Odds ratios and 95% CI, attributable risk, and population attributable risk were calculated. RESULTS: The percentage of SGA infants and preterm birth increased with higher levels of prenatal alcohol exposure; however, the association between alcohol intake and SGA infants was attenuated after adjustment for maternal smoking. Low levels of prenatal alcohol were not associated with preterm birth; however, binge drinking resulted in a nonsignificant increase in odds. Preterm birth was associated with moderate and higher levels of prenatal alcohol consumption for the group of women who ceased drinking before the second trimester. This group of women was significantly more likely to deliver a preterm infant than women who abstained from alcohol (adjusted OR 1.73 [95% CI 1.01 3.14]). CONCLUSIONS: Alcohol intake at higher levels, particularly heavy and binge drinking patterns, is associated with increased risk of preterm birth even when drinking is ceased before the second trimester. This finding, however, is based on small numbers and needs further investigation. Dose and timing of prenatal alcohol exposure appears to affect preterm delivery and should be considered in future research and health education. PMID- 19187373 TI - The maternal Mediterranean dietary pattern is associated with a reduced risk of spina bifida in the offspring. AB - OBJECTIVE: The objective of this study was to test the hypothesis whether a maternal dietary pattern is associated with the risk of spina bifida (SB) in the offspring. DESIGN: Case-control study. SETTING: Eight clinic sites in the Netherlands, 1999-2001. SAMPLE: A total of 50 mothers of children with SB and 81 control mothers. METHODS: Maternal food intakes were obtained by food frequency questionnaires at the standardised study moment of 14 months after the birth of the index child. Principal component factor analysis (PCA) and reduced rank regression (RRR) were used to identify dietary patterns. MAIN OUTCOME MEASURES: Maternal biomarkers were used as response measures in the RRR analysis and composed of serum and red blood cell (RBC) folate, serum vitamin B12 and total plasma homocysteine. The strength of the use of the dietary pattern in association with SB risk was estimated by odds ratios and 95% CI with the highest quartiles of the dietary pattern as reference. RESULTS: A predominantly Mediterranean dietary pattern was identified by both PCA and RRR. Those dietary patterns were highly correlated (r = 0.51, P < 0.001) and characterised by joint intakes of fruit, vegetables, vegetable oil, alcohol, fish, legumes and cereals and low intakes of potatoes and sweets. We observed a significantly increased risk of SB offspring in mothers with a weak use of the Mediterranean dietary pattern, OR 2.7 (95% CI 1.2-6.1) and OR 3.5 (95% CI 1.5-7.9). The Mediterranean dietary pattern was correlated with higher levels of serum and RBC folate, serum vitamin B12 and lower plasma homocysteine. CONCLUSION: The Mediterranean dietary pattern seems to be associated with reduction in the risk of offspring being affected by SB. PMID- 19187374 TI - High maternal vitamin E intake by diet or supplements is associated with congenital heart defects in the offspring. AB - OBJECTIVE: To study associations between maternal dietary and supplement intake of antioxidants vitamin E, retinol and congenital heart defects (CHDs). DESIGN: Case-control study. SETTING: Erasmus MC, University Medical Center Rotterdam, the Netherlands. POPULATION: Participants were 276 case mothers of a child with CHD and 324 control mothers with their children. METHODS: Food frequency questionnaires covering the intake of the previous 4 weeks were filled out at 16 months after the index pregnancy. Data were compared between cases and controls using the Mann-Whitney U test. Risk estimates for the association between CHD and dietary intake of vitamin E and retinol were estimated in a multivariable logistic regression model. MAIN OUTCOME MEASURES: Medians (5-95th percentile) and odds ratios with 95% CI. RESULTS: Dietary vitamin E intake was higher in case mothers than in controls, 13.3 (8.1-20.4) and 12.6 (8.5-19.8) mg/day (P= 0.05). CHD risk increased with rising dietary vitamin E intakes (P-trend = 0.01). Periconception use of vitamin E supplements in addition to a high dietary vitamin E intake above 14.9 mg/day up to nine-fold increased CHD risk. Retinol intakes were not significantly different between the groups and not associated with CHD risk. CONCLUSIONS: High maternal vitamin E by diet and supplements is associated with an increased risk of CHD offspring. PMID- 19187375 TI - Uterine and umbilical artery Doppler are comparable in predicting perinatal outcome of growth-restricted fetuses. AB - OBJECTIVE: To compare umbilical and uterine artery Doppler in predicting outcome of pregnancies suspected of fetal growth restriction (FGR). DESIGN: A prospective study included 353 singleton pregnancies complicated by an FGR fetus. SETTING: University Hospital setting. SAMPLE: Pregnancies suspected of FGR diagnosed by ultrasound fetal biometry during a 5-year period. MAIN OUTCOME MEASURE: Perinatal outcome in relation to uterine and umbilical artery Doppler. METHODS: The women underwent Doppler examination of the umbilical and uterine arteries. Results from the uterine, but not the umbilical artery, were blind to the woman and managing obstetrician. The Doppler results were related to perinatal outcomes including small for gestational age newborns, caesarean delivery, premature delivery (<37 weeks of gestation) and admission of the newborn to a neonatal intensive care unit. RESULTS: Abnormal uterine artery Doppler velocimetry was seen in 120 (33.4%) pregnancies and abnormal umbilical artery Doppler in 102 (28.4%). There was a statistically significant correlation between abnormal Doppler of both the umbilical and uterine arteries and adverse outcome of pregnancy. The two vessels were comparable in predicting adverse outcome. Women with normal umbilical artery Doppler (251) were analysed separately. Abnormal uterine artery Doppler, seen in 61 (24.3%) of those women, showed a statistically significant correlation for adverse outcome of pregnancy. CONCLUSIONS: Doppler examinations of the uterine and/or the umbilical arteries seem to be comparable as predictors of outcome in pregnancies complicated by FGR. Including uterine artery Doppler in the surveillance of growth-restricted fetuses might detect a group of pregnancies at high risk, even though the umbilical artery Doppler was normal. PMID- 19187376 TI - Evaluation of postpartum blood loss after misoprostol-induced labour. AB - OBJECTIVE: To objectively evaluate postpartum blood loss after successful misoprostol induction and compare it with blood loss after oxytocin induction of labour. DESIGN: Prospective randomised study. SETTING: Labour ward in university maternity hospital. POPULATION: A total of 150 women up to third parity with completed 40 weeks of singleton normal pregnancy, average size cephalic fetus. METHODS: Cases were randomised between oxytocin induction and misoprostol induction. Blood was collected in suction set and measured in the delivery room starting after delivery of the fetus and was evaluated by pad weighing in the following 6 hours. Pre- and postdelivery haematocrit were measured and difference between the two values was assessed and analysed. MAIN OUTCOME MEASURES: Success of induction, induction delivery interval, postpartum blood loss, and difference between pre- and postdelivery haematocrit. RESULTS: Induced labour was significantly faster with misoprostol induction (P < 0.001). Blood loss and haematocrit difference was significantly greater in the misoprostol group than in oxytocin group (P < 0.02 and 0.001, respectively). Blood loss in both groups was significantly correlated with higher initial Bishop score (P < 0.001 and 0.024, respectively) and short labour duration (P < 0.0002 and 0.0001, respectively). CONCLUSIONS: Misoprostol induction is associated with increased blood loss especially when used in women with high Bishop score; therefore, it is better reserved for cases requiring cervical ripening. PMID- 19187377 TI - Continuous versus interrupted sutures for repair of episiotomy or second-degree perineal tears: a randomised controlled trial. AB - OBJECTIVE: To evaluate the repair techniques of continuous and interrupted methods for episiotomy or perineal tears. DESIGN: A randomised controlled trial. SETTING: The Hospital Universitario Principe de Asturias, a state hospital belonging to the community of Madrid. SAMPLE: Four hundred forty-five women who had undergone vaginal deliveries with episiotomies or second-grade tearing of the perineum between September 2005 and July 2007. METHODS: One group was repaired with continuous, nonlocking sutures involving the vagina, perineum, and subcutaneous tissues. The other group had continuous, locking sutures of the vagina, interrupted sutures in the perineal muscles, and interrupted transcutaneous sutures. The threads used for stitching were identical in both groups. MAIN OUTCOME MEASURES: The participants were questioned regarding the sensation of pain and the use of painkillers on the second and the tenth days, and 3 months postpartum. RESULTS: When comparing the group with continuous suture to the group with interrupted sutures, the differences included less repair time (1 minute; P= 0.017) and less suture material used (relative risk [RR], 3.2, 95% CI: 2.6-4.0). The comparison of pain on the second and tenth days, and 3 months postpartum were not statistically different between the two techniques (RR, 1.08, 95% CI: 0.74-1.57; RR, 0.96, 95% CI: 0.59-1.55; and RR, 0.68, 95% CI: 0.19-2.46, respectively). CONCLUSIONS: Although we did not demonstrate that one technique was better than the other in the incidence of pain in the short or long term, we showed that episiotomy and perineal tear repairs with continuous suturing were quicker and used less suture material without an increase in complication than interrupted suturing. PMID- 19187378 TI - Body composition by dual-energy X-ray absorptiometry in women with previous pre eclampsia or small-for-gestational-age offspring. AB - OBJECTIVE: To investigate differences in body composition and fat distribution between women with previous pre-eclampsia or small-for-gestational-age (SGA) offspring and those with uncomplicated pregnancies. DESIGN: Cohort study. SETTING: Population-based study in a genetically isolated population in the southwest of the Netherlands. POPULATION: Women after pregnancies complicated by pre-eclampsia (n=45), SGA offspring (n=53) and uncomplicated pregnancies (n=106). METHODS: Women were compared for body composition and fat distribution variables, assessed by dual-energy X-ray absorptiometry (DXA) and anthropometrics at a mean follow-up time of 10.8 (SD +/- 5.9) years after pregnancy. MAIN OUTCOME MEASURES: Total lean and fat mass, android fat mass, gynoid fat mass, android-to-gynoid fat ratio, waist and hip circumference, waist-to-hip ratio. RESULTS: Women with previous pre-eclampsia compared with controls had higher mean total fat mass index (11.5 +/- 0.6 versus 9.7 +/- 0.4 kg/m(2); P = 0.03), lean mass index (15.8 +/- 0.3 versus 14.5 +/- 0.2 kg/m(2); P =0.001) and body mass index ([BMI]; 28.4 +/- 0.8 versus 25.4 +/- 0.5 kg/m(2); P = 0.005). Their waist circumferences (90.7 +/- 2.0 versus 78.5 +/- 1.3 cm; P < 0.001) and waist-to-hip ratios (0.86 +/- 0.01 versus 0.77 +/- 0.01; P < 0.001) were also higher as well as android fat mass (2.8 +/- 0.2 versus 2.1 +/- 0.1 kg; P = 0.01) and android-to-gynoid fat ratios (0.45 +/- 0.02 versus 0.39 +/- 0.01; P = 0.02). Mean total fat, lean and BMI was not significantly different between women with previous SGA offspring and controls, yet waist-to-hip ratios (0.83 +/- 0.01; P < 0.001) were higher. The observed differences in waist and hip circumference, waist-to-hip ratio and gynoid fat mass could not be attributed to differences in BMI. CONCLUSION: Women with previous pre-eclampsia or SGA offspring pregnancies compared with those with uncomplicated pregnancies have a preferential fat accumulation in the abdominal over hip region, which may explain, at least partly, their increased cardiovascular risk. PMID- 19187379 TI - Use of povidone-iodine during the first trimester of pregnancy: a correct practice? AB - Povidone-iodine (PVP-I) has been widely used as an antiseptic agent during invasive procedures for prenatal diagnosis. Women have been reported of thyroid dysfunction after simple exposure to PVP-I. We studied the effect on thyroid function and urinary iodine excretion after a single topical application of PVP-I in 31 women who had a miscarriage during the first trimester of pregnancy. PVP-I is absorbed through the skin and the vaginal mucosa, resulting in a sudden increase in the urinary excretion of iodine and a short-term variation in concentrations of thyroid hormones in maternal serum. This metabolic effect could have consequences for the embryo and the fetus during crucial stages of development. PMID- 19187380 TI - Uterine repair and successful pregnancy after myometrial and placental rupture with massive haemoperitoneum. PMID- 19187382 TI - Re: The relationship between sonographic fetal thymus size and the components of the systemic fetal inflammatory response syndrome in women with preterm prelabour rupture of membranes. PMID- 19187383 TI - Induction of labour. PMID- 19187386 TI - Re: Glove perforations during open surgery for gynaecological malignancies. PMID- 19187387 TI - Re: Effect of margin status on cervical intraepithelial neoplasia recurrence following LLETZ in women over 50 years. PMID- 19187388 TI - Posterior colpotomy-a retrieval route for solid ovarian tumours. PMID- 19187391 TI - Patient safety incidents associated with airway devices in critical care: a review of reports to the UK National Patient Safety Agency. AB - We used key words and letter sequences to identify airway-associated patient safety incidents submitted to the UK National Patient Safety Agency from critical care units in England and Wales. We identified 1085 such airway incidents submitted in the two years from October 2005 to September 2007. Three hundred and twelve incidents (28.8%) involved neonates or babies. Of the total 1085 incidents, 200 (18.4%) were associated with tracheal intubation, 53 (4.9%) with tracheostomy and 893 (82.3%) were post-procedure problems. One hundred and ten incidents (10.1%) were associated with more than temporary harm. Eighty-eight intubation incidents were associated with equipment problems. Partial displacement of tubes resulted in more than temporary harm to the patient more frequently than complete tube displacement (15.7% vs 3.8%). Capnography was not described in any cases of displacement or blockage of tracheal or tracheostomy tubes. Recommendations concerning minimum standards for capnography, availability and checking of equipment and tracheostomy placement are made. PMID- 19187392 TI - Generalized urticaria to fusidic acid. PMID- 19187393 TI - Early production of thymic stromal lymphopoietin precedes infiltration of dendritic cells expressing its receptor in allergen-induced late phase cutaneous responses in atopic subjects. AB - BACKGROUND: Thymic stromal lymphopoietin (TSLP) is an interleukin (IL)-7-like cytokine that triggers dendritic cell-mediated T helper (Th)2 inflammatory responses through a receptor consisting of a heterodimer of the IL-7 receptor alpha (IL-7Ralpha) chain and the TSLP receptor (TSLPR), which resembles the cytokine receptor common gamma chain. Dendritic cells activated by TSLP prime development of CD4(+) T cells into Th2 cells contributing to the pathogenesis of allergic inflammation. We hypothesized that allergen exposure induces expression of TSLP and results in recruitment of TSLPR bearing cells in the cutaneous allergen-induced late-phase reaction (LPR) in atopic subjects. METHODS: Skin biopsies were obtained from atopic subjects (n = 9) at various times after cutaneous allergen challenge. In situ hybridization and immunohistochemistry were used to determine TSLP mRNA expression and to measure infiltration of TSLPR(+) DC in skin LPR. RT-PCR and flow cytometry were employed to analyse TSLPR expression on isolated blood DC. RESULTS: Allergen-induced skin TSLP expression occurred as early as 1 h after allergen challenge, whereas TSLPR(+) and CD11c(+) cells infiltrated relatively late (24-48 h). The majority of TSLPR(+) cells were DC co expressing blood DC antigen-1 (BDCA-1) or BDCA-2. Freshly isolated blood DC expressed both TSLPR and IL-7Ralpha chains. Maturation and stimulation with TSLP or polyriboinosinic-polyribocytidylic acid in vitro upregulated the expression of both TSLPR and IL-7Ralpha chains in DC but not in chemoattractant receptor homologous molecule expressed on Th2 cells(+) CD4(+) T cells. CONCLUSION: The data suggest that TSLP plays a role in augmenting, through DC recruitment and activation, the development of Th2-type T cells in allergic inflammation. PMID- 19187394 TI - Increased number of offspring in first degree relatives of psychotic individuals: a partial explanation for the persistence of psychotic illnesses. AB - OBJECTIVE: As patients with psychotic illness have fewer offspring than controls, the persistence of psychotic illness is puzzling. We hypothesized that unaffected first-degree relatives of patients have more offspring than controls. METHOD: Probands were 4904, individuals with non-affective psychotic disorders identified from a hospitalization registry. Unaffected first degree relatives and matched controls were identified from the Israeli Population Registry. The number of offspring of unaffected parents, biological siblings and controls was ascertained. RESULTS: Unaffected parents of psychotic patients had more offspring/person than controls; 4.5 +/- 2.7 vs. 3.4 +/- 2.2, P = 0.000. Unaffected parents from familial psychosis families (more than one affected family member) had 1.83 more offspring than controls; unaffected parents from non familial psychosis families had 0.97 more offspring than controls (both P < 0.001). CONCLUSION: These findings might imply that genes which increase susceptibility for schizophrenia may be associated with increased number of offspring, perhaps supplying a partial explanation for the persistence of psychosis. PMID- 19187395 TI - Neonatal liver abscesses due to Candida infection effectively treated with caspofungin. AB - Candidiasis is relatively frequent in neonatal and pediatric intensive care units (ICUs), particularly in preterm infants less than 28 weeks of gestational age. Neonatal candidiasis shows high mortality and is often associated to poor neurodevelopmental prognosis in survivor patients. Amphotericin B and fluconazole are the first choice drugs for the treatment of neonatal candidiasis. Caspofungin is an alternative antifungal agent, which is recommended for invasive candidiasis in adults, but has been poorly experienced in neonates and infants as far as now. We report the first two infants with Candida liver abscesses treated with caspofungin. In the first infant bloodstream and liver lesions were cleared by combination therapy with fluconazole, liposomal amphotericin and caspofungin, while in the second one by caspofungin alone. CONCLUSION: Our observations confirm the efficacy and tolerability of caspofungin in the treatment of neonatal candidiasis refractory to conventional antifungal drugs. More extensive data are recommended in order to asses a specific neonatal schedule. PMID- 19187396 TI - Furosemide in preterm infants treated with indomethacin for patent ductus arteriosus. AB - OBJECTIVE: To evaluate the effect of furosemide on renal function and water balance in preterm infants treated with indomethacin (3 x 0.2 mg/kg at 12-h intervals) for symptomatic patent ductus arteriosus. PATIENTS AND METHODS: We performed a retrospective multi-centre double cohort study in preterm infants <32 weeks of gestational age. Thirty-two infants treated with furosemide (1 mg/kg i.v.) before each indomethacin dose (furosemide group) were matched with 32 infants with indomethacin treatment alone (control-group). Renal effects (urine output, weight gain, serum creatinine, sodium concentration) were registered. RESULTS: The study groups were comparable for gestational age, birth weight and day of therapy. Pretreatment differences were observed for urine output, weight and serum sodium. However, no differences were noticed in day-to-day urine output change or weight gain between the groups. A significant increase in serum creatinine concentration (50% vs. control, 18%; p < 0.05) and a concomitant significant decrease in serum sodium (-9 vs. control, -3 mmoL/L; p < 0.05) in the furosemide group was observed 72-96 h after starting therapy. CONCLUSION: Furosemide before each indomethacin dose resulted in a significant increase in serum creatinine and hyponatremia, without increasing urine output. PMID- 19187397 TI - Accuracy of Ottawa Ankle Rules to exclude fractures of the ankle and midfoot in children: a meta-analysis. AB - OBJECTIVES: The objectives were to conduct a systematic review to determine the diagnostic accuracy of the Ottawa Ankle Rules (OAR) to exclude ankle and midfoot fractures in children and the extent to which x-ray use could be reduced without missing significant fractures. METHODS: The authors conducted comprehensive searches of electronic databases and gray literature sources. Independent reviewers applied standard inclusion and exclusion criteria. The criterion standard diagnostic test was an ankle and/or foot x-ray or proxy measure to ensure no missed fractures. Standard 2 x 2 tables were constructed. Sensitivities and specificities were pooled using an approximation of the inverse variance; 95% confidence intervals (95% CIs) were calculated using the exact method. Likelihood ratios (LR +/-) and diagnostic odds ratios were combined under DerSimonian and Laird random effects model. RESULTS: A pooled analysis of 12 studies (N = 3,130) identified 671 fractures (prevalence = 21.4%). Ten studies reported Salter-Harris Type I (SH-I) fractures. The pooled sensitivity was 98.5% (95% CI = 97.3 to 99.2), suggesting that the OAR can be used to rule out a fracture. Four of 10 missed fractures were characterized: 1 SH-I, 1 SH-IV, and 2 "insignificant fractures" (either SH-I or avulsion fractures <3 mm). The pooled estimate for rate of x-ray reduction was 24.8% (95% CI = 23.3% to 26.3%; range = 5% to 44%). CONCLUSIONS: The OAR appear to be a reliable tool to exclude fractures in children greater than 5 years of age presenting with ankle and midfoot injuries. Employing the OAR would significantly decrease x-ray use with a low likelihood of missing a fracture. PMID- 19187398 TI - FXYD1, a modulator of Na,K-ATPase activity, facilitates female sexual development by maintaining gonadotrophin-releasing hormone neuronal excitability. AB - The excitatory tone to gonadotrophin-releasing hormone (GnRH) neurones is a critical component underlying the pubertal increase in GnRH secretion. However, the homeostatic mechanisms modulating the response of GnRH neurones to excitatory inputs remain poorly understood. A basic mechanism of neuronal homeostasis is the Na(+),K(+)-ATPase-dependent restoration of Na(+) and K(+) transmembrane gradients after neuronal excitation. This activity is reduced in a mouse model of Rett syndrome (RTT), a neurodevelopmental disorder in which expression of FXYD1, a modulator of Na(+),K(+)-ATPase activity, is increased. We now report that the initiation, but not the completion of puberty, is advanced in girls with RTT, and that, in rodents, FXYD1 may contribute to the neuroendocrine regulation of female puberty by modulating GnRH neuronal excitability. Fxyd1 mRNA abundance reaches maximal levels in the female rat hypothalamus by the fourth postnatal week of life (i.e., around the time when the mode of GnRH secretion acquires an adult pattern of release). Although Fxyd1 mRNA expression is low in the hypothalamus, approximately 50% of GnRH neurones contain Fxyd1 transcripts. Whole-cell patch recording of GnRH-EGFP neurones revealed that the neurones of Fxyd1-null female mice respond to somatic current injections with a lower number of action potentials than wild-type cells. Both the age at vaginal opening and at first oestrous were delayed in Fxyd1(-/-) mice, but adult reproductive capacity was normal. These results suggest that FXYD1 contributes to facilitating the advent of puberty by maintaining GnRH neuronal excitability to incoming transsynaptic stimulatory inputs. PMID- 19187399 TI - Loop diuretics can cause clinical natriuretic failure: a prescription for volume expansion. AB - Ultrafiltration enhances volume removal and weight reduction vs diuretics. However, their differential impact on total body sodium, potassium, and magnesium has not been described. Fifteen patients with congestion despite diuretic therapy had urine electrolytes measured after a diuretic dose. Ultrafiltration was initiated and ultrafiltrate electrolytes were measured. The urine sodium after diuretics (60+/-47 mmol/L) was less than in the ultrafiltrate (134+/-8.0 mmol/L) (P=.000025). The urine potassium level after diuretics (41+/-23 mmol/L) was greater than in the ultrafiltrate (3.7+/-0.6 mmol/L) (P=.000017). The urine magnesium level after diuretics (5.2+/-3.1 mg/dL) was greater than in the ultrafiltrate (2.9+/-0.7 mg/dL) (P=.017). In acute decompensated heart failure patients with congestion despite diuretic therapy, diuretics are poor natriuretics and cause significant potassium and magnesium loss. Ultrafiltration extracts more sodium while sparing potassium and magnesium. The sustained clinical benefits of ultrafiltration compared with diuretics may be partly related to their disparate effects on total body sodium, potassium, and magnesium, in addition to their differential efficacy of volume removal. PMID- 19187400 TI - Carvedilol produces sustained long-term benefits: follow-up at 12 years. AB - The authors measured long-term outcomes of patients who initiated carvedilol between 1990 and 1992 to test the hypothesis that carvedilol produces sustained benefits in heart failure patients. The study population consisted of 57 patients who completed a carvedilol placebo-controlled phase II trial. Patients were given open-label carvedilol and were titrated to the maximum dose. Patients were assessed by serial multigated acquisition, echocardiography, and symptom scores. Survival was assessed for all patients and censored as of January 1, 2004. Survival for ischemic vs nonischemic patients was compared using the log-rank test and further compared using Cox regression, controlling for covariates. Etiology of heart failure was ischemic in 15 patients and nonischemic in 42 patients. Median follow-up was 12.9 years. Resting left ventricular ejection fraction (LVEF) and heart failure symptom scores improved at 4 months of treatment and were sustained at 24 months. Left ventricular internal diameter in systole (LVIDS) and left ventricular internal diameter in diastole decreased significantly at 4 and 8 months, respectively, and LVIDS continued to improve at 24 months. Overall mortality was 43% in nonischemic patients and 73% in ischemic patients. In a multivariate analysis, ischemic etiology and baseline LVEF were significant predictors of mortality. Carvedilol produces sustained improvements in left ventricular remodeling and symptoms. Long-term survival is good, particularly in nonischemic patients. PMID- 19187401 TI - Initial emergency department systolic blood pressure predicts left ventricular systolic function in acute decompensated heart failure. AB - Ejection fraction (EF) is often unknown in patients who present with acute decompensated heart failure (ADHF). The objective of this study was to determine whether a patient's systolic blood pressure is associated with their left ventricular EF. This study was a retrospective chart review of all patients admitted to an emergency department (ED) observation unit from January 2002 to December 2004. A low EF was defined as <40%. Among 475 patients, the median age was 72 years, 53% were men, 40% were white, 59% were black, and 59% had a low EF. Patients with low EFs were more likely male ( P<.0001), with prior congestive heart disease ( P<.0001), longer QRS duration ( P<.0001), left bundle branch block ( P<.0001), and higher B-type natriuretic peptide ( P<.0001). The low EF group was less likely to have diabetes ( P<.0001). Adjusted odds ratios for an EF >or=40% were significant at all systolic blood pressure readings >120 mm Hg. Having an ED systolic BP >120 mm Hg is associated with significantly higher rates of preserved left ventricular systolic function in patients with ADHF. PMID- 19187402 TI - Optimization of cardiac resynchronization therapy: echocardiographic vs semiautomatic device algorithms. AB - Large evidence supports the importance of individualized optimization of cardiac resynchronization therapy in patients with congestive heart failure. The aim of this study was to compare a recently developed intracardiac electrogram (IEGM) based method with the Doppler echocardiographic (ECHO)-based method to calculate optimal atrioventricular (AV) and interventricular (VV) delays. Ten male patients implanted with a St Jude Medical resynchronization device received AV and VV delay assessment with both the IEGM and the ECHO-based methods. Estimates of the optimal AV and VV delays assessed by the 2 tested methods proved highly comparable. No difference emerged between the IEGM (126.8+/-22.7) and the ECHO (127.3+/-19.8) AV delay values (P=.987). The VV delay suggested by ECHO was highly significantly correlated with the delays calculated by the IEGM method (35+/-27.6 vs 21.31+/-24.31; r(2)=0.78; P<.001). These preliminary data support the evidence that an IEGM based cardiac resynchronization optimization method may be as reliable as a complete ECHO assessment. PMID- 19187403 TI - Determination of left ventricular systolic dyssynchrony by tissue Doppler echocardiography in patients with nonischemic dilated cardiomyopathy. AB - The authors investigated the left ventricular systolic dyssynchrony (LVSD) parameters in nonischemic dilated cardiomyopathy (DCM) by tissue Doppler echocardiography. A total of 56 DCM patients with normal sinus rhythm and 15 controls were recruited. The patients were subgrouped according to QRS duration (QRSd): QRSd 120 milliseconds (group II). Ts was defined as the duration from the initiation of QRS to the peak systolic velocity. Patients with DCM had longer intraventricular delay time (IVDT) (101.8 vs 47.4 milliseconds; P<.0001) and standard deviation of Ts (Ts-SD) values (34.9 vs 17.8 milliseconds; P<.0001) compared with controls. There was no difference in IVDT and Ts-SD between the 2 DCM subgroups. The cut-off value for LVSD was 60 milliseconds for IVDT and 25 milliseconds for Ts-SD. The prevalence of LVSD in DCM patients was found to be 78% in group I and 94% in group II. LVSD is common in DCM, regardless of QRSd. PMID- 19187404 TI - Atrial fibrillation and heart failure. AB - Atrial fibrillation (AF) is the most common arrhythmia in congestive heart failure (HF) and indicates a worse prognosis. AF increases HF symptoms and increases in prevalence with increasing New York Heart Association class. AF also interferes with the ideal management of HF. Across all HF etiologies, AF may be a marker of disease severity. Yet, controversies exist regarding whether strategies to restore and maintain sinus rhythm can improve outcomes in HF. It is also unclear what the optimal strategy is to suppress the ventricular response to AF in patients with HF. As HF incidence and prevalence continue to rise, the authors sought to reinvestigate current literature that relates AF to HF and examine the impact of therapy on HF and/or AF. The authors performed a literature review using a MEDLINE search from 1966 to the present and included existing literature based on their strength of evidence. PMID- 19187405 TI - Do comorbid psychiatric disorders contribute to the pathogenesis of tako-tsubo syndrome? A review of pathogenesis. AB - Patients with tako-tsubo cardiomyopathy have unusual characteristics, including a disproportionate occurrence in postmenopausal women and a strong association with mental and physical stress. These patients present with chest pain, abnormal electrocardiographic findings, and myocardial enzyme leaks. They appear to have acute coronary syndrome but have normal epicardial coronary vessels and reversible left ventricular dysfunction. High circulating levels of catecholamines likely cause this syndrome through direct injury of myocardial cells. The frequent occurrence in postmenopausal women has not been explained. This association might reflect changes in autonomic control of the cardiovascular system and in baroreceptor sensitivity in postmenopausal women. Women in this age group also have a high frequency of both depression and anxiety. These 2 psychiatric diagnoses are associated with episodic symptoms, including acute panic attacks, which can be associated with catecholamine release in the heart. Therefore, tako-tsubo cardiomyopathy may occur in postmenopausal women as a consequence of comorbid psychiatric diagnoses and normal age-related changes in autonomic control of the cardiovascular system. This hypothesis warrants prospective case-control studies. PMID- 19187406 TI - Intra-aortic balloon counterpulsation for the treatment of myocardial infarction complicated by acute severe heart failure. AB - Intractable heart failure (HF) remains a leading fatal complication of acute myocardial infarction (AMI). Intra-aortic balloon pump (IABP) counterpulsation assists the failing left ventricle and accelerates the functional recovery of stunned myocardium. Despite its remarkable performance, the beneficial effects of the IABP in the setting of acute HF or cardiogenic shock complicating AMI have not been confirmed in a randomized clinical trial. Instead, large amounts of information have been collected in observational studies or in retrospective analyses of randomized trials of reperfusion strategies in patients with AMI. The strategy of "stabilize with IABP, treat with reperfusion, and transfer for complete revascularization" has, thus far, yielded the best outcomes, and every effort should be made to implement this strategy in all patients presenting with AMI and severe HF. PMID- 19187407 TI - Peak oxygen consumption and heart failure prognosis--does race, sex, or fat explain the discrepancy? PMID- 19187408 TI - Cardiac carcinoid. PMID- 19187409 TI - Tako-tsubo cardiomyopathy presenting with acute pulmonary edema. PMID- 19187410 TI - History of heart failure. PMID- 19187411 TI - Scheduled bright light for treatment of insomnia in older adults. AB - OBJECTIVES: To determine whether bright light can improve sleep in older individuals with insomnia. DESIGN: Single-blind, placebo-controlled, 12-week, parallel-group randomized design comparing four treatment groups representing a factorial combination of two lighting conditions and two times of light administration. SETTING: At-home light treatment; eight office therapy sessions. PARTICIPANTS: Thirty-six women and fifteen men (aged 63.6+/-7.1) meeting primary insomnia criteria recruited from the community. INTERVENTION: A 12-week program of sleep hygiene and exposure to bright ( approximately 4,000 lux) or dim light ( approximately 65 lux) scheduled daily in the morning or evening for 45 minutes. MEASUREMENTS: Within-group changes were observed for subjective (sleep logs, questionnaires) and objective (actigraphy, polysomnography) sleep measures after morning or evening bright light. RESULTS: Within-group changes for subjective sleep measures after morning or evening bright light were not significantly different from those observed after exposure to scheduled dim light. Objective sleep changes (actigraphy, polysomnography) after treatment were not significantly different between the bright and dim light groups. Scheduled light exposure was able to shift the circadian phase predictably but was unrelated to changes in objective or subjective sleep measures. A polymorphism in CLOCK predicted morningness but did not moderate the effects of light on sleep. The phase angle between the circadian system (melatonin midpoint) and sleep (darkness) predicted the magnitude of phase delays, but not phase advances, engendered by bright light. CONCLUSION: Except for one subjective measure, scheduled morning or evening bright light effects were not different from those of scheduled dim light. Thus, support was not found for bright light treatment of older individuals with primary insomnia. PMID- 19187413 TI - Inconsistency in classification and reporting of in-hospital falls. AB - OBJECTIVES: To investigate agreement between hospital staff on what constitutes a fall and should be recorded on an incident report, to identify factors that influence whether a scenario is classified as a fall, and to examine the effect of providing a definition of a fall on interrater agreement. DESIGN: Two interrater agreement trials with pre-postintervention design. SETTING: Seven hospitals. PARTICIPANTS: Four hundred forty-six hospital staff (nursing 76%, physiotherapy 14%, occupational therapy, 6%) working on participating wards over a 24-hour period. MEASUREMENTS: Fourteen video scenarios were played before participants, who were asked whether they would classify each scenario as a fall and whether they would complete an incident report in that scenario. A definition of a fall was then provided and the video scenarios replayed. Two definitions were investigated; the World Health Organization definition and a custom definition. RESULTS: Consensus was not evident in five of the 14 scenarios examined. Falls onto surfaces higher than the ground were less likely to be recorded on incident reports. Hospital and ward type influenced whether a scenario was classified as a fall. Overall agreement in classifying scenarios as a fall was only marginally greater after provision of the definition in both phases, although neither had an effect on whether staff would complete an incident report. CONCLUSION: Disagreement between hospital staff in what constitutes a fall may contribute to inconsistent research findings in this field and is not meaningfully improved through provision of a definition. PMID- 19187412 TI - Long-term function in an older cohort--the cardiovascular health study all stars study. AB - OBJECTIVES: To evaluate shared and unique risk factors for maintaining physical and cognitive function into the ninth decade and beyond. DESIGN: Longitudinal cohort study. SETTING: Four U.S. communities. PARTICIPANTS: One thousand six hundred seventy-seven participants in the Cardiovascular Health Study All Stars Study, assessed in 2005/06. Median age was 85 (range 77-102), 66.5% were women, and 16.6% were black. MEASUREMENTS: Intact function was defined as no difficulty with any activities of daily living and a score of 80 or higher on the Modified Mini-Mental State Examination. Baseline characteristics assessed in 1992/93 included demographics, behavioral health factors, chronic disease history, subclinical disease markers, cardiovascular risk factors, and inflammatory markers. Multinomial logistic regression was used to compare risk for physical disability, cognitive impairment,and combined impairments with no functional impairment. RESULTS: Of the 1,677 participants evaluated in both domains, 891 (53%) were functionally intact. Continuous measures of function, including the Digit Symbol Substitution Test and gait speed, showed that all groups, including the most functional, had declined over time. The functional group had less decline but also tended to have higher starting values. Functional individuals had a higher baseline health profile than those with either or cognitive impairment or both impairments combined. Women and individuals with greater weight had higher rates of physical impairment but not cognitive impairment. Risk factors common to both types of impairment included cardiovascular disease and hypertension. CONCLUSION: Intact function was found in only approximately half of these older adults in the ninth decade and beyond. High baseline function and low vascular disease risk characterized functional aging. PMID- 19187414 TI - Systematic review of measures of clinical significance employed in randomized controlled trials of drugs for dementia. AB - The ability to define thresholds for the clinical significance (clinical importance) of outcome measures in dementia drug research is critical to determining the changes on outcome measures that patients and families would consider worth the cost and worth risking the side effects of medications (i.e., clinical importance is central to informed consent when starting such medications). Thresholds for clinical significance are also required for drug trial sample size calculation (if the studies are to be powered to detect clinically important difference) and for decisions regarding whether medications should be accepted on formularies and should be funded. To better understand what measures of clinical significance have been employed in dementia drug research, a systematic review was performed of double-blind randomized controlled trials (RCTs) of cholinesterase inhibitors and memantine in subjects with Alzheimer's disease, vascular dementia, mixed dementia, and mild cognitive impairment. Of the 57 dementia drug RCTs reviewed, only 46% discussed the clinical significance of their results. The most commonly cited measures of clinical significance were a 4 point change in the Alzheimer's Disease Assessment Scale--Cognitive Subscale and changes on global scales. The majority of measures of clinical significance were opinion based. Only one study empirically measured patient perspective regarding thresholds for clinical significance. Despite being central to the interpretation of trial results and to decisions regarding whether to employ trial findings in clinical practice, patient- and caregiver-centered measures of clinical significance have not been adequately studied and integrated into dementia drug RCTs. It is recommended that discussions of clinical importance receive greater emphasis in research standards published by organizations such as the CONSORT group (http://www.consort-statement.org). Drug formulary review committees and licensing agencies (e.g., U.S. Food and Drug Administration, European Agency for the Evaluation of Medicinal Products, Health Canada) should consider requiring an assessment of clinical significance of the drugs they review. To move this field forward, funding agencies should consider initiating requests for proposals focused on the determination of the minimally clinically important difference (MCID) of outcome measures employed in dementia research. Once empirical data on MCIDs are available, then these funding agencies should consider supporting a consensus conference to review and select the optimal measures of clinical importance in dementia research. A preliminary organizational framework of measures of clinical significance is presented in this article to facilitate the work of such a consensus forum. PMID- 19187415 TI - Respiratory syncytial virus outbreak in a long-term care facility detected using reverse transcriptase polymerase chain reaction: an argument for real-time detection methods. AB - OBJECTIVES: To report an outbreak of respiratory synctyial virus (RSV) in a long term care facility (LTCF) during ongoing routine respiratory illness surveillance. DESIGN: Rapid antigen testing, viral culture, direct fluorescent antibody (DFA) testing, and reverse transcriptase polymerase chain reaction (RT PCR) testing for up to 15 viruses in symptomatic residents and chart review. SETTING: A 120-bed LTCF. MEASUREMENTS: Comparison of rapid antigen testing, respiratory viral cultures, and DFA testing and RT-PCR in residents with symptoms of a respiratory tract infection. RESULTS: Twenty-two of 52 residents developed symptoms of a respiratory tract infection between January 29, 2008, and February 26, 2008. RSV was detected using RT-PCR in seven (32%) of the 22 cases. None of the seven cases had positive RSV rapid antigen testing, and only two had positive culture or DFA results. This outbreak occurred during a time when state wide RSV rates were rapidly declining. One patient was admitted to the hospital during the infection and subsequently died. CONCLUSION: RSV may cause outbreaks in LTCFs that traditional diagnostic methods do not detect. RT-PCR can provide a more timely and accurate diagnosis of outbreaks, which allows for early symptomatic treatment, rational use of antibiotics, and improved infection control. PMID- 19187416 TI - The co-occurrence of chronic diseases and geriatric syndromes: the health and retirement study. AB - OBJECTIVES: To analyze the co-occurrence, in adults aged 65 and older, of five conditions that are highly prevalent, lead to substantial morbidity, and have evidence-based guidelines for management and well-developed measures of medical care quality. DESIGN: Secondary data analysis of the 2004 wave of the Health and Retirement Study (HRS). SETTING: Nationally representative health interview survey. PARTICIPANTS: Respondents in the 2004 wave of the HRS aged 65 and older. MEASUREMENTS: Self-reported presence of five index conditions (three chronic diseases (coronary artery disease, congestive heart failure, and diabetes mellitus) and two geriatric syndromes (urinary incontinence and injurious falls)) and demographic information (age, sex, race, living situation, net worth, and education). RESULTS: Eleven thousand one hundred thirteen adults, representing 37.1 million Americans aged 65 and older, were interviewed. Forty-five percent were aged 76 and older, 58% were female, 8% were African American, and 4% resided in a nursing home. Respondents with more conditions were older and more likely to be female, single, and residing in a nursing home (all P<.001). Fifty-six percent had at least one of the five index conditions, and 23% had two or more. Of respondents with one condition, 20% to 55% (depending on the index condition) had two or more additional conditions. CONCLUSION: Five common conditions (3 chronic diseases, 2 geriatric syndromes) often co-occur in older adults, suggesting that coordinated management of comorbid conditions, both diseases and geriatric syndromes, is important. Care guidelines and quality indicators, rather than considering one condition at a time, should be developed to address comprehensive and coordinated management of co-occurring diseases and geriatric syndromes. PMID- 19187417 TI - Long-term effect on mortality of a home intervention that reduces functional difficulties in older adults: results from a randomized trial. AB - OBJECTIVES: To evaluate the long-term mortality effect of a home-based intervention previously shown to reduce functional difficulties and whether survivorship benefits differ according to initial mortality risk level. DESIGN: Two-group randomized trial with survivorship followed up to 4 years from study entry. SETTING: Homes of urban community-living elderly people. PARTICIPANTS: Three hundred nineteen adults aged 70 and older with difficulties performing daily activities. INTERVENTION: Occupational and physical therapy sessions to instruct participants in compensatory strategies, home modifications, home safety, fall recovery techniques, and balance and muscle strength exercises. MEASUREMENTS: Survival time was number of days between baseline interview and date of death, as determined using data from the National Death Index or December 31, 2005. Participants were stratified according to baseline mortality risk (low, moderate, high) using a prognostic indicator. RESULTS: At 2 years, intervention participants (n=160) had a 5.6% mortality rate (n=9 deaths) and controls (n=159) a 13.2% rate (n=21 deaths; P=.02). Mortality rates remained lower for intervention participants up to 3.5 years from study entry. At 2 years, intervention participants with moderate mortality risk had a 16.7% mortality rate (n=16 deaths/96), compared with 28.2% for equivalent control group participants (n=24 deaths/85; P=.02). By 3 years, mortality rates were not statistically significantly different between the experimental and control groups. CONCLUSIONS: The intervention extended survivorship up to 3.5 years and maintained statistically significant differences for 2 years. Subjects at moderate mortality risk derived the most intervention benefit. Findings suggest that the intervention could be a low-cost clinical tool to delay functional decline and mortality. PMID- 19187418 TI - Responsiveness and clinical utility of the geriatric self-efficacy index for urinary incontinence. AB - OBJECTIVES: To report on the responsiveness testing and clinical utility of the 12-item Geriatric Self-Efficacy Index for Urinary Incontinence (GSE-UI). DESIGN: Prospective cohort study. SETTING: Six urinary incontinence (UI) outpatient clinics in Quebec, Canada. PARTICIPANTS: Community-dwelling incontinent adults aged 65 and older. MEASUREMENTS: The abridged 12-item GSE-UI, measuring older adults' level of confidence for preventing urine loss, was administered to all new consecutive incontinent patients 1 week before their initial clinic visit, at baseline, and 3 months posttreatment. At follow-up, a positive rating of improvement in UI was ascertained from patients and their physicians using the Patient's and Clinician's Global Impression of Improvement scales, respectively. Responsiveness of the GSE-UI was calculated using Guyatt's change index. Its clinical utility was determined using receiver operating curves. RESULTS: Eighty nine of 228 eligible patients (39.0%) participated (mean age 72.6+5.8, range 65 90). At 3-month follow-up, 22.5% of patients were very much better, and 41.6% were a little or much better. Guyatt's change index was 2.6 for patients who changed by a clinically meaningful amount and 1.5 for patients having experienced any level of improvement. An improvement of 14 points on the 12-item GSE-UI had a sensitivity of 75.1% and a specificity of 78.2% for detecting clinically meaningful changes in UI status. Mean GSE-UI scores varied according to improvement status (P<.001) and correlated with changes in quality-of-life scores (r=0.7, P<.001) and reductions in UI episodes (r=0.4, P=.004). CONCLUSION: The GSE-UI is responsive and clinically useful. PMID- 19187419 TI - The nursing home minimum data set for vision and its association with visual acuity and contrast sensitivity. AB - OBJECTIVES: To evaluate the association between the Minimum Data Set's (MDS) Vision Patterns section and near and distance visual acuity and contrast sensitivity in nursing home residents. DESIGN: Cross-sectional study. SETTING: Seventeen nursing homes in the Birmingham, Alabama, area. PARTICIPANTS: Three hundred seventy-one nursing home residents aged 55 and older with Mini-Mental State Examination scores of 13 or greater. MEASUREMENTS: The MDS 2.0 assessment for vision from the date closest to acuity and contrast sensitivity assessment were obtained from the resident's medical record. Acuity and contrast sensitivity were measured using the ETDRS chart and Pelli-Robson chart, respectively. RESULTS: The MDS rating of visual status was associated with distance and near visual acuity and contrast sensitivity. The MDS performed poorly in distinguishing residents with mild visual impairment from those with moderate visual impairment. For residents who were rated on the MDS as having adequate vision, 45.9% had distance acuity worse than 20/40 in the better eye, 72.8% had near acuity worse than 20/40 in the better eye, and 85.8% had contrast sensitivity less than 1.50. CONCLUSION: The MDS 2.0 assessment for vision in nursing home residents is positively associated with visual acuity and contrast sensitivity but does not adequately distinguish between individuals with mild and moderate visual impairment and classifies many as having adequate vision who have visual impairment. The validity of the MDS 2.0 as a mechanism for triggering comprehensive eye care for nursing home residents is questionable. PMID- 19187420 TI - Working across boundaries to improve health outcomes: a case study of a housing support and outreach service for homeless people living with HIV. AB - This paper reports the findings of an evaluation of the 'Housing Support, Outreach and Referral' service developed to support people living with HIV who were homeless or at risk of homelessness. The service was set up as part of the Supporting People Health Pilot programme established to demonstrate the policy links between housing support services and health and social care services by encouraging the development of integrated services. The paper considers the role of housing support in improving people's health, and considers the challenges of working across housing, health and social care boundaries. The evaluation of the health pilot employed two main sources of data collection: quarterly project evaluation reports, which collected process data as well as reporting progress against aims and objectives, and semi-structured interviews with professionals from all key stakeholder groups and agencies, and with people who used services. Over the course of 15 months, 56 referrals were received of which 27 were accepted. Fifteen people received tenancy support of whom 12 were helped to access temporary accommodation. At the end of the 15 months, all of the tenancies had been maintained. In addition, 18 people registered with a general practitioner and 13 registered with an HIV clinic. Interviews with professionals emphasised the importance of the local joint working context, the involvement of the voluntary sector and the role of the support workers as factors that accounted for these outcomes. Those using services placed most emphasis on the flexibility of the support worker role. Importantly, interviews with professionals and those using services suggest that the role of support worker incorporates two dimensions--those of networker/navigator as well as advocate- and that both dimensions are important in determining the effectiveness of the service. PMID- 19187421 TI - Elderly women's experiences of living with fall risk in a fragile body: a reflective lifeworld approach. AB - The purpose of this qualitative study was to explore the lived experience of fall risk from a lifeworld perspective in elderly women with previous fragility fractures. Thirteen elderly women with a high risk of fall and fracture, aged 76 86, living in their own homes in rural areas, were recruited from a voluntary fracture prevention programme. All women had a history of fragility fractures and were interviewed in their homes from spring to autumn 2004. A phenomenological reflective lifeworld approach was chosen to analyse in-depth interview data. The study was conducted within an interdisciplinary research group inspired by dialogical research. Elderly women's life space has been narrowed due to advanced age, physical injury or by efforts to prevent new injuries leading to changes in self-perception. However, the women seek strategies to challenge limitations and insecurity, and strive to retain mobility and daily life routines. The four major constituents of the phenomenon 'elderly women's experiences of fall risk' emerged in this study: a changing body, living with precaution, ambiguous dependency and influence and need for understanding. Employing the women's thoughts and resources in trust-based dialogues with caregivers may strengthen their concord and the prospects to continue an active life. Elderly women seek strategies to challenge limitations and feelings of insecurity, and strive to maintain mobility and daily life routines. A trust-based care respecting the preferences of the women seemed to stimulate behavioural change in maintaining an active life. PMID- 19187422 TI - The process of decision-making in home-care case management: implications for the introduction of universal assessment and information technology. AB - Increasingly, jurisdictions are adopting universal assessment procedures and information technology to aid in healthcare data collection and care planning. Before their potential can be realised, a better understanding is needed of how these systems can best be used to support clinical practice. We investigated the decision-making process and information needs of home-care case managers in Ontario, Canada, prior to the widespread use of universal assessment, with a view of determining how universal assessment and information technology could best support this work. Three focus groups and two individual interviews were conducted; questioning focused on decision-making in the post-acute care of individuals recovering from a hip fracture. We found that case managers' decisional process was one of a clinician-broker, combining clinical expertise and information about local services to support patient goals within the context of limited resources. This process represented expert decision-making, and the case managers valued their ability to carry out non-standardised interviews and override system directives when they noted that data may be misleading. Clear information needs were found in four areas: services available outside of their regions, patient medical information, patient pre-morbid functional status and partner/spouse health and functional status. Implications for the use of universal assessment are discussed. Recommendations are made for further research to determine the impact of universal assessment and information technology on the process and outcome of home-care case manager decision-making. PMID- 19187423 TI - Designing sexual health services for young people: a methodology for capturing the user voice. AB - The aim of the study was to assess the suitability of the Thurstone paired comparison method for capturing the user voice, through a survey of young people's views on the most salient priorities for a sexual health service. A convenience sample of 161 12-24 year olds was used. A psychometrically robust questionnaire was developed from a review of the relevant literature and from the information provided by three focus groups. The data derived from both stages were distilled into seven themes, and adapted to a Thurstone paired comparison format, in which each theme was paired with every other theme, with an 8-point scale between each pairing (21 pairings in total). Respondents were required to indicate their preference for one theme over the other in each pairing. The questionnaire was completed by 161 young people between April and July 2007, and the results were analysed using the Kendall coefficient of concordance to establish the degree of within-group agreement. The results suggested that there was significant agreement as to the essential desirable features of a sexual health service, both within the whole sample as well as within sub-samples (i.e. gender, age group and previous sexual health service use). The priorities were privacy, and a dedicated service close to home, with a drop-in facility and male and female staff being next most important, and an informal service and young staff being lowest priorities. The feedback from the pilot study, the 40% return and absence of spoiled questionnaires together indicated that the respondents found the method acceptable, while the actual findings corroborated those from other studies. Taken together, these results suggest that the Thurstone method offers a quick and simple method of capturing the user voice, with the results having sufficient validity to inform the planning of a local sexual health service. PMID- 19187425 TI - Tribute to Bill Narayan. PMID- 19187426 TI - AIDS pathogenesis: a tale of two monkeys. AB - Despite many years of intense scientific effort, the pathogenic mechanisms underlying the immunodeficiency that follows human immunodeficiency virus (HIV) infection are still poorly understood. This lack of understanding is likely the main reason why at present there is neither a cure nor a vaccine for AIDS. Important clues on the immunopathogenesis of primate lentiviral infections have been provided by comparative studies of two simian models of SIV infection: the non-pathogenic SIV infection of sooty mangabey, an African natural host species, and the pathogenic SIV-infection of non-natural host rhesus macaques, that develop a disease that closely resembles AIDS in humans. While the final mechanisms underlying the difference in infection outcome between these two species are still incompletely understood, a series of recent studies has allowed the identification of key similarities and differences between the two models of infection. In this article we summarize these findings and review the main implications in terms of HIV pathogenesis, therapy, and vaccines. PMID- 19187428 TI - Protection afforded by live attenuated SIV is associated with rapid killing kinetics of CTLs. AB - BACKGROUND: Live attenuated SIV vaccines are highly efficacious, but how they mediate protection is poorly understood. A feature of the effectiveness of live attenuated vaccines is their ability to control high dose challenge viruses early, without a large peak of acute viraemia. We hypothesized that long-lived antigen exposure from live attenuated SIV may result in CD8+ cytotoxic T lymphocytes persistently capable of rapidly cytolytic potential. METHODS: We employed a kinetic degranulation assay to study multiple tetramer+ SIV-specific CTL specificities before and after the SIV(mac251) challenge of pigtail macaques inoculated with a live attenuated SIV. RESULTS: Live attenuated SIV-vaccinated animals rapidly controlled a subsequent challenge, with minimal viraemia after exposure. For over 9 months after the initial vaccination with live attenuated SIV we could detect both Gag- and Tat-specific CTLs that maintained a long-term capacity to rapidly degranulate (CD107a expression) and release granzyme B within 30 minutes of antigen exposure. This rapid cytolytic phenotype was maintained throughout the early period after challenge, despite the absence of a marked enhancement in CTL frequencies. CONCLUSIONS: Our results suggest that highly functional CTLs may contribute to the remarkable efficacy of live attenuated SIV vaccines. Studying the killing kinetics of CTLs induced by other, safer, HIV vaccines could facilitate a better understanding of the requirements for an effective HIV vaccine. PMID- 19187427 TI - Differential pathogenicity of SHIV infection in pig-tailed and rhesus macaques. AB - BACKGROUND: Differential pathogenicity has been observed in cynomolgus and rhesus macaques following primate lentivirus infection. However, little is known about the comparative susceptibility of pig-tailed macaques to lentivirus infection and diseases. METHODS: We compared the in vivo infectivity and pathogenicity of a CCR5-tropic SHIV(SF162 P4) after intravenous, intravaginal or intrarectal inoculation in rhesus and pig-tailed macaques. Plasma viral load, peripheral blood CD4(+) T cell counts and clinical signs were monitored. RESULTS: Both rhesus and pig-tailed macaques are similarly susceptible to SHIV(SF162 P4) infection by intravenous and mucosal routes. However, infection was significantly more robust in pig-tailed macaques than in rhesus, resulting in persistent viremia in 9/21 pig-tails vs. 2/24 rhesus (P < 0.013) and severe CD4(+) T-cell depletion in 2/21 pig-tails (vs. none in rhesus). CONCLUSIONS: Together with earlier observations, our findings underscore the importance of considering host genetic and immunological factors when comparing vaccine efficacy in different macaque species. PMID- 19187429 TI - Kinetics of T lymphocyte apoptosis and the cellular immune response in SIVmac239 infected rhesus macaques. AB - BACKGROUND: Although increased apoptosis is a central feature of AIDS, little is known about its kinetics or relationship to the early host response in acute HIV/SIV infection. METHODS: Ex vivo apoptosis in freshly isolated peripheral blood and lymph node lymphocytes was monitored longitudinally in SIVmac239 infected rhesus macaques by flow-cytometric detection of active caspase-3, cleaved poly (ADP-ribose) polymerase, and fragmented DNA. RESULTS: Increased apoptosis of multiple lymphocyte subsets was observed in the first 2 weeks following SIV infection. Apoptosis of CD4+ T lymphocytes was of low magnitude but peaked earlier than other T lymphocyte subsets. A 10- to 36-fold increase in CD8+ T lymphocyte apoptosis coincided temporally with onset of the SIV-specific cellular immune response and enrichment of caspase-3-positive cells within recently proliferating, activated CD8+ T lymphocytes. CONCLUSIONS: The virus specific T lymphocyte response to primary infection and generalized non-specific immune activation contribute to the pathogenesis of apoptosis in acute SIV infection. PMID- 19187431 TI - Loss of growth factor receptor signaling in the oral mucosa during primary SIV infection may enhance apoptosis and promote pathogenesis. AB - BACKGROUND: The development of susceptibility to secondary pathogenic infections in the oral cavity during HIV infection is likely to result from or coincide with deterioration of the local mucosal immune system. METHODS: We have utilized the SIV model to investigate the kinetics and magnitude of oral pathogenesis during systemic dissemination of intravenously inoculated SIVmac251. RESULTS: Viral replication was detected in oropharyngeal lymph nodes at 6 weeks post-infection and shown to be coincident with a broad scale loss of growth factor receptor transcription in the oral mucosa, providing multiple avenues for blocking the normal activity of apoptosis inhibitors that function through Bcl2- and p53 dependent pathways. CONCLUSIONS: Our findings suggest that the normal balance between cell death and regeneration may be rapidly disrupted in the oral mucosa during the early stages of immunodeficiency virus infection, setting the stage for continuing deterioration of immune function and the development of susceptibility to secondary infections. PMID- 19187430 TI - Association between decreased CXCL12 and CCL25 expression and increased apoptosis in lymphoid tissues of cynomolgus macaques during SIV infection. AB - BACKGROUND: Chemokines likely play multiple roles in HIV-1 and SIV pathogenesis. To examine potential associations between chemokine expression levels and apoptosis of cells in lymphoid tissues during SIV infection, we measured chemokine and cytokine mRNA levels in multiple lymphoid tissues compartments from uninfected and SIV-infected cynomolgus macaques (Macaca fascicularis). METHODS: Real-time RT-PCR was used to measure host mRNA levels in macaque lymphoid tissues. Proliferating or apoptotic cells were identified in lymphoid tissues by immunohistochemistry. RESULTS: We found that CXCL12 and CCL25 mRNAs in SIV infected lymphoid tissues were decreased and their levels were negatively correlated with the numbers of proliferating and apoptotic cells. In vitro analyses revealed that CXCL12 and CCL25 were capable of reducing apoptosis induced by SIV infection. CONCLUSIONS: These findings suggest that increased apoptosis in lymphoid tissues due to reduced levels of anti-apoptotic chemokines might be a mechanism that contributes to loss of immune function following pathogenic SIV infection. PMID- 19187432 TI - CLTA-4 blockade in vivo promotes the generation of short-lived effector CD8 T cells and a more persistent central memory CD4 T cell response. AB - BACKGROUND: Previously, we examined the effects of in vivo CTLA-4 blockade using a fully human monoclonal antibody as a part of a DNA vaccination regimen in cynomolgus macaques (Macaca fascicularis). We observed that while the antibody had little effect on the IFN-gamma ELISpot response, CTLA-4 blockade enhanced antigen-specific cellular proliferation in both CD4(+) and CD8(+)T-cell compartments. METHODS: We examine the specific effects of CTLA-4 blockade on memory T-cell compartments following the third immunization and 10 months following a fourth immunization, during the memory phase of the immune response. RESULTS: CLTA-4 blockade enhanced CD4(+) and CD8(+) central memory (CD28(hi), CD95(hi)) T-cell responses as well as a short-lived CD8(+) effector (CD28(lo), CD95(hi)) T-cell response. CONCLUSIONS: These data suggest differing effects of CTLA-4 blockade on CD4(+) and CD8(+) T cells with implications on the clinical use of anti-CTLA-4 antibodies for enhancement of vaccine strategies or treatment of human disease. PMID- 19187433 TI - Safety, immunogenicity and efficacy of peptide-pulsed cellular immunotherapy in macaques. AB - BACKGROUND: Simple and effective delivery methods for cellular immunotherapies are needed. We recently published on the effectiveness of using ex vivo pulsing of overlapping SIV Gag 15mer peptides onto fresh peripheral blood cells in 32 SIV(mac251)-infected pigtail macaques. METHODS: We now report on the safety of this approach, analysis of a novel assay for immunogenicity, the effect of an MHC allele, Mane-A*10, on CD8 T cell escape occurring and disease outcome. RESULTS: The vaccine strategy was safe, with no perturbations in weight or hematological profiles in comparison to controls. The high levels of SIV-specific T cell immunogenicity of this approach was confirmed using a novel assay measuring upregulation of surface CD134 of CD4 T cells. A substantial effect of the Mane A*10 allele in reducing SIV viral load of pigtail macaques was observed in both vaccinees and controls; the virologic efficacy of the immunotherapy in comparison to controls was greatest in Mane-A*10- animals. Escape mutations at several new CD8 T cell epitopes throughout the SIV proteome were observed, primarily in animals with poorer virologic control. CONCLUSIONS: In summary, we provide further information that peptide-pulsed PBMC are a safe, immunogenic and effective immunotherapy. The observed influence of MHC alleles and immune escape allows us to design more insightful future immunotherapy studies. PMID- 19187434 TI - Baboon carboxylesterases 1 and 2: sequences, structures and phylogenetic relationships with human and other primate carboxylesterases. AB - BACKGROUND: Carboxylesterase (CES) is predominantly responsible for the detoxification of a wide range of drugs and narcotics, and catalyze several reactions in cholesterol and fatty acid metabolism. Studies of the genetic and biochemical properties of primate CES may contribute to an improved understanding of human disease, including atherosclerosis, obesity and drug addiction, for which non-human primates serve as useful animal models. METHODS: We cloned and sequenced baboon CES1 and CES2 and used in vitro and in silico methods to predict protein secondary and tertiary structures, and examined evolutionary relationships for these enzymes with other primate and mouse CES orthologs. RESULTS AND CONCLUSIONS: We found that baboon CES1 and CES2 proteins retained extensive similarity with human CES1 and CES2, shared key structural features reported for human CES1, and showed family specific sequences consistent with their multimeric and monomeric subunit structures respectively. PMID- 19187435 TI - A novel Brucella isolate in association with two cases of stillbirth in non-human primates - first report. AB - BACKGROUND: Brucellosis is veterinary and human health problem. METHODS: A 13 year-old wild caught multiparous and an 8-year-old colony-born nulliparous baboon had stillbirths in the second trimester of pregnancy. Culture isolates from both postpartum uteruses were characterized using traditional biochemical analysis, PCR, and multilocus sequencing. RESULTS: The isolates morphologically resembled Brucella although their phenotypic characteristics were not consistent with any currently described species. The isolates represent a novel lineage within the genus with unique alleles, not previously seen in surveys of greater than 300 isolates representing the known diversity of the genus, present at 5/9 loci examined. CONCLUSIONS: The described cases are to the best of our knowledge the first presentation of a naturally acquired Brucella infection in non-human primates associated with stillbirths from the same colony where Brucella seropositivity in the baboons was described 45 years ago. The organism appears to represent a previously undescribed Brucella species. PMID- 19187437 TI - An immunohistochemical and stereological analysis of PSI-induced nigral neuronal degeneration in the rat. AB - Systemic administration of the proteasomal inhibitor I (PSI) to rats was reported to cause progressive nigral dopaminergic neuronal loss but this is disputed. A major controversy centres over the use of manual counting of tyrosine hydroxylase (TH) positive neurons at the level of third cranial nerve as opposed to employing systematic stereological analysis of cell loss in the entire substantia nigra (SN). To provide a method of marking SN neurones independent of protein expression, fluorogold (FG) was stereotaxically injected bilaterally into the striatum of male Wistar rats to retrogradely label nigral dopaminergic neurons. After 1 week, animals were treated with six doses of PSI (8 mg/kg, s.c.) or its vehicle (dimethyl sulphoxide) on alternate days over a 2-week period. Five weeks after the last treatment, PSI-treated animals showed decreased spontaneous locomotor activity and reduced TH positive SN cell number at the level of the third cranial nerve compared to control rats. Manual cell counting showed loss of FG-labelled SN neurones at this level, with a subpopulation of surviving neurons displaying abnormal morphology. Manual counting of all FG-labelled cells in the entire SN also showed regional PSI-induced loss of neurones with both normal and compromised morphology. Stereological optical fractionator estimates of total FG labelled cell number confirmed the manual cell counting data both at the level of the third cranial nerve and throughout the entire SN. These findings confirm that PSI does cause a persistent nigral dopaminergic neuronal loss. The reason for the lack of reproducibility between laboratories requires further investigation. We suggest that a failure to distinguish between TH-positive neurones with normal and abnormal morphology following PSI administration contributes to equivocal results. PMID- 19187436 TI - The basal flux of Akt in the mitochondria is mediated by heat shock protein 90. AB - Akt is a known client protein of heat shock protein 90 (HSP90). We have found that HSP90 is responsible for Akt accumulation in the mitochondria in unstimulated cells. Treatment of SH-SY5Y neuroblastoma cells and human embryonic kidney cells with the HSP90 inhibitors novobiocin and geldanamycin caused substantial decreases in the level of Akt in the mitochondria without affecting the level of Akt in the cytosol. Moreover, intracerebroventricular injection of novobiocin into mice brains decreased Akt levels in cortical mitochondria. Knockdown of HSP90 expression with short interfering RNA also caused a significant decrease in Akt levels in the mitochondria without affecting total Akt levels. Using a mitochondrial import assay it was found that Akt is transported into the mitochondria. Furthermore, it was found that the mitochondrial import of Akt was independent of Akt activation as both an unmodified Akt and constitutively active mutant Akt; both readily accumulated in the mitochondria in an HSP90-dependent manner. Interestingly, incubation of isolated mitochondria with constitutively active Akt caused visible alterations in mitochondrial morphology, including pronounced remodeling of the mitochondrial matrix. This effect was blocked when Akt was mostly excluded from the mitochondria with novobiocin treatment. These results indicate that the level of Akt in the mitochondria is dependent on HSP90 chaperoning activity and that Akt import can cause dynamic changes in mitochondrial configuration. PMID- 19187438 TI - Estradiol induces physical association of neuronal nitric oxide synthase with NMDA receptor and promotes nitric oxide formation via estrogen receptor activation in primary neuronal cultures. AB - Estrogens and nitric oxide (NO) exert wide-ranging effects on brain function. Recent evidence suggested that one important mechanism for the regulation of NO production may reside in the differential coupling of the calcium-activated neuronal NO synthase (nNOS) to glutamate NMDA receptor channels harboring NR2B subunits by the scaffolding protein post-synaptic density-95 (PSD-95), and that estrogens promote the formation of this ternary complex. Here, we demonstrate that 30-min estradiol-treatment triggers the production of NO by physically and functionally coupling NMDA receptors to nNOS in primary neurons of the rat preoptic region in vitro. The ability of estradiol to activate neuronal NO signaling in preoptic neurons and to promote changes in protein-protein interactions is blocked by ICI 182,780, an estrogen receptor antagonist. In addition, blockade of NMDA receptor NR2B subunit activity with ifenprodil or disruption of PSD-95 synthesis in preoptic neurons by treatment with an anti sense oligodeoxynucleotide inhibited the estradiol-promoted stimulation of NO release in cultured preoptic neurons. Thus, estrogen receptor-mediated stimulation of the nNOS/PSD-95/NMDA receptor complex assembly is likely to be a critical component of the signaling process by which estradiol facilitates coupling of glutamatergic fluxes for NO production in neurons. PMID- 19187439 TI - A new culturing strategy improves functional neuronal development of human neural progenitor cells. AB - Cell replacement therapies that rely on in vitro differentiation of human neural progenitor cells are a promising strategy to compensate the progressive cell loss in neurodegenerative disorders like Parkinson's disease. We and others observed, that the functional differentiation of progenitors in standard differentiation medium remains limited. The aim of the present study was to optimize neuronal in vitro differentiation by mimicking the physiological shift from depolarizing to hyperpolarizing conditions that occurs during early brain development. Differentiation was initiated using a depolarizing high potassium- and low sodium containing medium. Subsequently, the high potassium-containing medium was replaced by a hyperpolarizing medium containing low potassium and high sodium concentrations. This two-phase strategy significantly promoted the expression of neuronal markers, enhanced neurite growth, enlarged sodium inward currents, and increased action potential firing. Thus, depolarizing followed by hyperpolarizing culture conditions enable developing human neural progenitor cells to adopt more mature functional qualities. PMID- 19187440 TI - Differential effects of albumin on microglia and macrophages; implications for neurodegeneration following blood-brain barrier damage. AB - Microglial activation by blood-borne factors following blood-brain barrier damage may play a significant role in subsequent neuropathogenesis of several neurodegenerative diseases. Exposure of primary cultured rat brain microglia to pure, fatty acid- and lipid-deficient rat serum albumin or fraction V, (fatty acid and lipid-containing rat serum albumin), caused inducible nitric oxide synthase (iNOS) expression, glutamate release, tumour necrosis factor alpha (TNFalpha) and transforming growth factor-beta1 release. iNOS expression was attenuated by the MAPK/extracellular signal-regulated kinase pathway inhibitor U0126 and the phosphorylated forms of extracellular signal-regulated kinase 1 and 2 were detectable in microglia treated with albumin or fraction V. Glutamate release was prevented by l-alpha-aminoadipate and glutathione levels in microglia rose on exposure to albumin. Conditioned medium from microglia exposed to albumin or fraction V was neurotoxic. Peripheral macrophages were resistant to the effects of albumin but both microglia and macrophages responded to lipopolysaccharide, which induced interleukin-1 beta and tumour necrosis factor alpha release, cyclooxygenase-2 and iNOS expression in both cell types, indicating a discrete desensitised pathway in macrophages for albumin which was not desensitised in microglia. Thus, exposure of microglia in the brain to albumin may contribute to neuronal damage following blood-brain barrier breakdown and point to resident microglia rather than infiltrating macrophages as therapeutic targets. PMID- 19187441 TI - p53-dependent control of cell death by nicastrin: lack of requirement for presenilin-dependent gamma-secretase complex. AB - Nicastrin (NCT) is a component of the presenilin (PS)-dependent gamma-secretase complexes that liberate amyloid beta-peptides from the beta-Amyloid Precursor Protein. Several lines of evidence indicate that the members of these complexes could also contribute to the control of cell death. Here we show that over expression of NCT increases the viability of human embryonic kidney (HEK293) cells and decreases staurosporine (STS)- and thapsigargin (TPS)-induced caspase-3 activation in various cell lines from human and neuronal origins by Akt-dependent pathway. NCT lowers p53 expression, transcriptional activity and promoter transactivation and reduces p53 phosphorylation. NCT-associated protection against STS-stimulated cell death was completely abolished by p53 deficiency. Conversely, the depletion of NCT drastically enhances STS-induced caspase-3 activation and p53 pathway and favored p53 nuclear translocation. We examined whether NCT protective function depends on PS-dependent gamma-secretase activity. First, a 29-amino acid deletion known to reduce NCT-dependent amyloid beta peptide production did not affect NCT-associated protective phenotype. Second, NCT still reduces STS-induced caspase-3 activation in fibroblasts lacking PS1 and PS2. Third, the gamma-secretase inhibitor DFK167 did not affect NCT-mediated reduction of p53 activity. Altogether, our study indicates that NCT controls cell death via phosphoinositide 3-kinase/Akt and p53-dependent pathways and that this function remains independent of the activity and molecular integrity of the gamma secretase complexes. PMID- 19187442 TI - Densin-180: revised membrane topology, domain structure and phosphorylation status. AB - Densin-180 is a core component of post-synaptic densities, the highly complex molecular assemblies that mediate signaling between neuronal cells. It is a multi domain scaffold protein characterized by multiple leucine-rich repeat domains plus a single Psd95/Discs large/Zona occludens-1 domain. In its original topology model a single transmembrane segment was proposed with an extracellular N terminus and an intracellular C-terminus. However, recently discovered in vivo phosphorylation sites are incompatible with this topology. Here, we discuss an all-intracellular and membrane-associated localization of Densin-180 that is consistent with and supported by all the latest experimental data. This revised topology which now includes also a phosphorylation-rich area will have deciding influence on future research involving Densin-180 and its signaling. PMID- 19187443 TI - Aminopeptidase A contributes to the N-terminal truncation of amyloid beta peptide. AB - Several lines of data previously indicated that N-terminally truncated forms of amyloid-beta (Abeta) peptides are likely the earliest and more abundant species immunohistochemically detectable in Alzheimer's disease-affected brains. It is noteworthy that the free N-terminal residue of full-length Abeta (fl-Abeta) is an aspartyl residue, suggesting that Abeta could be susceptible to exopeptidasic attack by aminopeptidase A (APA)-like proteases. In this context, we have examined whether APA could target Abeta peptides in both cell-free and cellular models. We first show that the general aminopeptidase inhibitor amastatin as well as two distinct aminopeptidase A inhibitors EC33 and pl302 both significantly increase the recovery of genuine fl-Abeta peptides generated by cells over expressing Swedish-mutated beta amyloid precursor protein (APP) while the aminopeptidase N blocker pl250 did not modify fl-Abeta recovery. In agreement with this observation, we establish that over-expressed APA drastically reduces, in a calcium dependent manner, fl-Abeta but not APP IntraCellular Domain in a cell-free model of Abeta production. In agreement with the above data, we show that recombinant APA degrades fl-Abeta in a pl302-sensitive manner. Interestingly, we also show that EC33 and pl302 lower staurosporine-stimulated activation of caspase-3 in wild-type fibroblasts but not in betaAPP/beta-amyloid precursor protein-like protein 2 (APLP2) double knockout fibroblasts, suggesting that protecting endogenous fl-Abeta physiological production triggers neuroprotective phenotype. By contrast, EC33 does not modify staurosporine induced caspase-3 activation in wild-type and Swedish-mutated betaAPP-HEK293 expressing cells that display exacerbated production of Abeta. Overall, our data establish that APA contributes to the N-terminal truncation of Abeta and suggest that this cleavage is likely abrogating a protective function associated with physiological but not supraphysiological levels of genuine fl-Abeta peptides. PMID- 19187444 TI - Lipid rafts regulate 2-arachidonoylglycerol metabolism and physiological activity in the striatum. AB - Several G protein-associated receptors and synaptic proteins function within lipid rafts, which are subdomains of the plasma membranes that contain high concentrations of cholesterol. In this study we addressed the possible role of lipid rafts in the control of endocannabinoid system in striatal slices. Disruption of lipid rafts following cholesterol depletion with methyl-beta cyclodestrin (MCD) failed to affect synthesis and degradation of anandamide, while it caused a marked increase in the synthesis and concentration of 2 arachidonoylglycerol (2-AG), as well as in the binding activity of cannabinoid CB1 receptors. Surprisingly, endogenous 2-AG-mediated control of GABA transmission was not potentiated by MCD treatment and, in contrast, neither basal nor 3,5-Dihydroxyphenylglycine-stimulated 2-AG altered GABA synapses in cholesterol-depleted slices. Synaptic response to the cannabinoid CB1 receptor agonist HU210 was however intact in MCD-treated slices, indicating that reduced sensitivity of cannabinoid CB1 receptors does not explain why endogenous 2-AG is ineffective in inhibiting striatal GABA transmission after cholesterol depletion. Confocal microscopy analysis suggested that disruption of raft integrity by MCD might uncouple metabotropic glutamate 5-CB1 receptor interaction by altering the correct localization of both receptors in striatal neuron elements. In conclusion, our data indicate that disruption of raft integrity causes a complex alteration of the endocannabinoid signalling in the striatum. PMID- 19187445 TI - Alterations of myelin-specific proteins and sphingolipids characterize the brains of acid sphingomyelinase-deficient mice, an animal model of Niemann-Pick disease type A. AB - Niemann-Pick disease (NPD) type A is a neurodegenerative disorder caused by sphingomyelin (SM) accumulation in lysosomes relying on reduced or absent acid sphingomyelinase (ASM) activity. NPD-A patients develop progressive neurodegeneration including cerebral and cerebellar atrophy, relevant Purkinje cell and myelin deficiency with death within 3 years. ASM'knock-out' (ASMKO) mice, an animal model of NPD-A, develop a phenotype largely mimicking that of NPD A. The mechanisms underlying myelin formation are poorly documented in ASMKO mice. In this study we determined the content of four myelin-specific proteins, myelin basic protein (MBP), 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNP), myelin associated glycoprotein (MAG) and proteolipid protein (PLP), and that of myelin-enriched sphingolipids in the brains of ASMKO and wild-type mice in early stages of post-natal (pn) life. Protein and mRNA analysis revealed that in ASMKO mice beginning from 4 post-natal weeks (wk-pn), the expression levels of MAG, CNP, and MBP were below those observed in wild-type mice and the same applied to PLP at 10 wk-pn. Moreover, at 4 wk-pn the expression of SOX10, one of the transcription factors involved in oligodendrocyte development and maintenance was lower in ASMKO mice. Lipid analysis showed that SM and the gangliosides GM3 and GM2 accumulated in the brains of ASMKO mice, as opposed to galactocerebroside and galactosulfocerebroside that, in parallel with the mRNAs of UDP-galactose ceramide galactosyltransferase and galactose-3-O-sulfotransferase 1, the two transferases involved in their synthesis, decreased. Myelin lipid analysis showed a progressive sphingomyelin accumulation in ASMKO mice; noteworthy, of the two sphingomyelin species known to be resolved by TLC, only that with the lower Rf accumulated. The immunohistochemical analysis showed that the reduced expression of myelin specific proteins in ASMKO mice at 10 wk-pn was not restricted to the Purkinje layer of the cerebellar cortex but involved the cerebral cortex as well. In conclusion, reduced oligodendrocyte metabolic activity is likely to be the chief cause of myelin deficiency in ASMKO mice, thus shedding light on the molecular dysfunctions underlying neurodegeneration in NPD-A. PMID- 19187446 TI - Reduction of protein kinase C zeta inhibits migration and invasion of human glioblastoma cells. AB - Glioblastomas are the most aggressive forms of primary brain tumors with their tendency to invade surrounding healthy brain tissues, rendering them largely incurable. In this report, we used small-interference RNA technology to knock down the expression of protein kinase C (PKC) zeta, which resulted in specific and massive impairment of glioblastoma cell migration and invasion. We also explained the fundamental molecular processes of glioblastoma migration and invasion in which PKCzeta is a participant. The silence of PKCzeta expression likewise impaired the phosphorylation of LIN-11, Isl1 and MEC-3 protein domain kinase (LIMK) and cofilin, which is a critical step in cofilin recycling and actin polymerization. Consistent with the defects in cell adhesion, phosphorylation of integrin beta1 was also dampened. Therefore, PKCzeta regulated both cytoskeleton rearrangement and cell adhesion, which contributed to cell migration. Additionally, there was down-regulation of matrix metalloprotease-9 expression in siPKCzeta/LN-229 cells, which coincided with decreased invasion both in vitro and in vivo. These results indicate that PKCzeta is involved in the control of glioblastoma cell migration and invasion by regulating the cytoskeleton rearrangement, cell adhesion, and matrix metalloprotease-9 expression. Collectively, these findings suggest that PKCzeta is a potential therapeutic target for glioblastoma infiltration. PMID- 19187447 TI - Activation of pre-synaptic M-type K+ channels inhibits [3H]D-aspartate release by reducing Ca2+ entry through P/Q-type voltage-gated Ca2+ channels. AB - In this study, the functional consequences of the pharmacological modulation of the M-current (I(KM)) on cytoplasmic Ca(2+) intracellular Ca(2+)concentration ([Ca(2+)](i)) changes and excitatory neurotransmitter release triggered by various stimuli from isolated rat cortical synaptosomes have been investigated. K(v)7.2 immunoreactivity was identified in pre-synaptic elements in cortical slices and isolated glutamatergic cortical synaptosomes. In cerebrocortical synaptosomes exposed to 20 mM [K(+)](e), the I(KM) activator retigabine (RT, 10 microM) inhibited [(3)H]D-aspartate ([(3)H]D-Asp) release and caused membrane hyperpolarization; both these effects were prevented by the I(KM) blocker XE-991 (20 microM). The I(KM) activators RT (0.1-30 microM), flupirtine (10 microM) and BMS-204352 (10 microM) inhibited 20 mM [K(+)](e)-induced synaptosomal [Ca(2+)](i) increases; XE-991 (20 microM) abolished RT-induced inhibition of depolarization triggered [Ca(2+)](i) transients. The P/Q-type voltage-sensitive Ca(2+)channel (VSCC) blocker omega-agatoxin IVA prevented RT-induced inhibition of depolarization-induced [Ca(2+)](i) increase and [(3)H]D-Asp release, whereas the N-type blocker omega-conotoxin GVIA failed to do so. Finally, 10 microM RT did not modify the increase of [Ca(2+)](i) and the resulting enhancement of [(3)H]D Asp release induced by [Ca(2+)](i) mobilization from intracellular stores, or by store-operated Ca(2+)channel activation. Collectively, the present data reveal that the pharmacological activation of I(KM) regulates depolarization-induced [(3)H]D-Asp release from cerebrocortical synaptosomes by selectively controlling the changes of [Ca(2+)](i) occurring through P/Q-type VSCCs. PMID- 19187448 TI - The effects of substance use on offender crime scene behavior. AB - Substance use has an effect on an individual's propensity to commit acquisitive crime with recent studies showing substance users more likely to leave forensic material at a crime scene. An examination of acquisitive crime solved in Northamptonshire, U.K., during 2006 enabled 70 crime scene behavior characteristics to be analyzed for substance and nonsubstance use offenders. Logistical regression analyses have identified statistically significant crime scene behavior predictors that were found to be either present at or absent from the crime scene when the offender was a substance user. Most significant predictors present were indicative of a lack of preparation by the offender, irrational behavior, and a desire to steal high value, easily disposed of, property. Most significant predictors absent from the crime scene were indicative of more planning, preparation, and execution by the offender. Consideration is given to how this crime scene behavior might be used by police investigators to identify offenders. PMID- 19187449 TI - Toxicant exposure and mental health--individual, social, and public health considerations. AB - Thoughts and moods are the result of biological processes; disordered thoughts and moods may be the result of disordered biological processes. As brain dysfunction can manifest with emotional symptoms or behavioral signs, the etiology of some mental health afflictions and some abnormal conduct is pathophysiological rather than pathopsychological. Various studies confirm that some chemical toxicants which modify brain physiology have the potential to affect mood, cognitive function, and to provoke socially undesirable outcomes. With pervasive concern about myriad chemical agents in the environment and resultant toxicant bioaccumulation, human exposure assessment has become a clinically relevant area of medical investigation. Adverse exposure and toxicant body burden should routinely be explored as an etiological determinant in assorted health afflictions including disordered thinking, moods, and behavior. The impact of toxicant bioaccumulation in a patient with neuropsychiatric symptoms is presented for consideration as an example of the potential benefit of recognizing and implementing exposure assessment. PMID- 19187450 TI - Degeneration of nuclei and mitochondria in human hairs. AB - It is generally accepted that nuclei degrade in developing hair shafts but the point at which such occurs has not been investigated. The fate of mitochondria in the keratinizing hair shaft has been less clear. This study uses transmission electron microscopy to investigate when nuclei and mitochondria are no longer visible in the developing hair shaft. Serial sections were obtained from anagen head hairs absent follicles in order to determine the sequence of degradation of nuclei and mitochondria in the hair shaft by starting at the root bulb and proceeding toward the hair tip. It was demonstrated that nuclei and mitochondria become invisible in the keratinizing hair shaft at about the same time. This was found to occur fairly early in the process at the level of the hair shaft where the hair cuticle becomes permanent. PMID- 19187451 TI - GrinLine identification using digital imaging and Adobe Photoshop. AB - The purpose of this study was to outline a method by which an antemortem photograph of a victim can be critically compared with a postmortem photograph in an effort to facilitate the identification process. Ten subjects, between 27 and 55 years old provided historical pictures of themselves exhibiting a broad smile showing anterior teeth to some extent (a grin). These photos were termed "antemortem" for the purpose of the study. A digital camera was used to take a current photo of each subject's grin. These photos represented the "postmortem" images. A single subject's "postmortem" photo set was randomly selected to be the "unknown victim." These combined data of the unknown and the 10 antemortem subjects were digitally stored and, using Adobe Photoshop software, the images were sized and oriented for comparative analysis. The goal was to devise a technique that could facilitate the accurate determination of which "antemortem" subject was the "unknown." The generation of antemortem digital overlays of the teeth visible in a grin and the comparison of those overlays to the images of the postmortem dentition is the foundation of the technique. The comparisons made using the GrinLine Identification Technique may assist medical examiners and coroners in making identifications or exclusions. PMID- 19187452 TI - Insect successional pattern of a corpse in cooler months of subtropical southeastern Texas. AB - Here, we characterize the cool weather insect fauna found associated with partially skeletonized and desiccated human remains recovered from an abandoned house in an urban area of subtropical, coastal Galveston County, Texas, and use the information to conclude an approximate postmortem interval of 7-10 months. The predominant factors that allow for a confident assessment of the postmortem interval include climatological data, entomological data, and anthropological data. The documented insect fauna represents a unique assemblage present in a particular environment (an urban abandoned house in coastal Texas) at a particular time of year (winter) and includes expected forensically significant insects such as calliphorid flies, muscid flies, and dermestid beetles but also includes less commonly encountered insects such as an unusually dense population of live case-making clothes moths. PMID- 19187453 TI - Identification and comparison of electrical tapes using instrumental and statistical techniques: II. Organic composition of the tape backing and adhesive. AB - The microtexture and elemental composition of the backing of electrical tapes have been shown to be highly discriminating. In this study, the organic composition of electrical tape was evaluated as a complementary means of distinguishing tape brands. The backing and adhesive of 72 rolls of electrical tape were analyzed via Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy (ATR FTIR) and discriminant analysis was used to classify all samples by brand. Generally, the accuracy for FTIR data (88-99%) was higher than that for elemental data (86-94%). FTIR spectra from the adhesive layer were the most discriminating. In separate studies, two fragments of blast-damaged tape were correctly assigned to their brand of origin and discriminant analysis was used to quantitatively associate or exclude tape samples from two bombing cases. PMID- 19187454 TI - Fluorescence and structural degradation in composite resins as a function of temperature. AB - Detecting composite resin upon postmortem examination can be difficult. Ultraviolet illumination has been suggested to ease location of this material; however, this may not be advisable in incineration situations. Understanding of the chemical and physical properties of resin as a function of temperature is an important parameter in identification of this material in incineration circumstances. Twenty-seven discs of resin, Quixx (Dentsply), Filtek Supreme (3 M), and Tetric Ceram (Ivoclar) were prepared and exposed to increasing heating conditions of 200 degrees C-900 degrees C in 100 degrees C increments for 30 min. Analysis was performed with Fourier transform infrared spectroscopy, ultraviolet visible light spectrophotometry, scanning electron microscopy/energy dispersive X ray spectroscopy, optical microscopy, and UV illumination. Characterization of the material occurred at each temperature range. The organic components and the fluorescence properties were lost at temperatures above 300 degrees C. The inorganic component remained through 900 degrees C. This information can aid in detection of resin in high temperature circumstances. PMID- 19187455 TI - "Theory of mind" and executive functioning in forensic patients with schizophrenia. AB - Previous studies in forensic patients with schizophrenia have shown that delinquent patients may outperform nondelinquent patients with regards to "theory of mind" (ToM). Findings were, however, confounded by a lack of control for executive functioning. We examined 33 forensic patients with schizophrenia regarding ToM, intelligence, executive functioning, and psychopathology. Results were compared with a nonforensic schizophrenia sample and a group of healthy controls. Both patient groups performed more poorly on most measures compared with controls. Forensic and nonforensic patients did not differ in task performance. In the forensic group ToM correlated inversely with "excitement" and cognitive symptoms. When "excitement" was covaried out, forensic patients outperformed nonforensic patients with regards to ToM. This study supports the hypothesis that schizophrenic patients with a criminal record are equally impaired in their ability to infer mental states compared with nonforensic patients, but for different reasons associated with a divergent psychopathological profile. PMID- 19187456 TI - A multi-drug intoxication fatality involving Xyrem (GHB). AB - Gamma-hydroxybutyrate (GHB) is best known as a recreational depressant drug, whose use has also been implicated in drug facilitated sexual assault cases. It is also available as a therapeutic agent (Xyrem) used for the treatment of daytime sleepiness or cataplexy associated with narcolepsy. This is a report of a case of a 53-year-old woman undergoing treatment with Xyrem for narcolepsy. The decedent was also prescribed tramadol, gabapentin, cetirizine, modafinil, carisoprodol, and Xyrem. Toxicological analysis of the blood revealed GHB 165.6 mg/L, and 90.7 mg/L in the urine. Blood GHB concentrations in the range 156-260 mg/L have been reported to induce moderately sound sleep. The combined use of central nervous system depressant drugs, together with her problematic sleep apnea, and snoring (both contraindications for GHB use) were determined to have caused this subject's death. The manner of death was determined to be accidental. PMID- 19187457 TI - Fathers who kill their children: an analysis of the literature. AB - Roughly half of filicidal acts are committed by fathers, though the majority of the literature focuses on maternal filicide. This paper reviews the existing literature on paternal filicide with the goal of identifying characteristics common among these fathers. Fathers who killed their children were, on average, in their mid thirties. The mean age of their victims was five. They may have multiple victims. Sons and daughters were killed in equal numbers. Reasons included death related to abuse, mental illness (including psychosis and depression), and revenge against a spouse. The method often involved wounding violence. Suicide following the act occurred frequently. After being tried for their crimes, filicidal fathers were more frequently incarcerated than hospitalized. Given the range of those capable of this act, mental health professionals must be alert to the possibility of filicide in a variety of fathers. Considering this risk, clinicians should inquire about thoughts of harming children, partners, and themselves. PMID- 19187458 TI - Preparation of tool mark standards with jewelry modeling waxes. AB - This paper presents how jewelry modeling waxes are used in the preparation of tool mark standards from exemplar tools. We have previously found that jewelry modeling waxes are ideal for preparing test tool marks from exemplar tools. In this study, simple methods and techniques are offered for the replication of accurate, highly detailed tool mark standards with jewelry modeling waxes. The techniques described here demonstrate the conditioning and proper use of jewelry modeling wax in the production of tool mark standards. The application of each test tool's working surface to a piece of the appropriate wax in a manner consistent with the tool's design is clearly illustrated. The resulting tool mark standards are exact, highly detailed, 1:1, negative impressions of the exemplar tool's working surface. These wax models have a long shelf life and are suitable for use in microscopic examination comparison of questioned and known tool marks. PMID- 19187459 TI - Concordance between the AmpFlSTR MiniFiler and AmpFlSTR Identifiler PCR amplification kits in the Kuwaiti population. AB - The AmpFlSTR MiniFiler polymerase chain reaction amplification kit, developed and supplied by Applied Biosystems, complements the AmpFlSTR Identifiler polymerase chain reaction amplification kit (Applied Biosystems, Warrington, U.K.) by improving the success rate when profiling DNA that is degraded or contains inhibitors. Before applying the MiniFiler kit to casework, the profiles from 200 unrelated Kuwaitis were compared to Identifiler profiles. Concordance was observed for 99.875% (1598 of 1600) of the compared STR loci. The two discordant profiles displayed allelic dropout: one at the D13S317 locus due to nonamplification of allele 10 in the MiniFiler profile, and one at the D18S51 locus due to nonamplification of allele 18 in the Identifiler profile. PMID- 19187460 TI - Estimating age of majority on third molars developmental stages in young adults from Thailand using a modified scoring technique. AB - The aim of this study was to achieve a referral database for dental age estimation of unaccompanied minors of Thai nationality. A total of 1199 orthopantomograms were collected from original Thai women and men equally divided in age categories between 15 and 24 years. On the radiographs, the developmental stage of the third molars was scored applying a modified scoring technique. Inter and intra-observer reliabilities were tested using kappa statistics. Correlation between the scores of all four wisdom teeth and left-right symmetry were evaluated with Pearson's correlation coefficient. Student's t-test on asymmetry was performed and regression formulas were calculated. The present database was the first to assemble third molar developmental scores on radiographs of Thai individuals and provides more appropriate dental age estimation of unaccompanied Thai minors. Future research on similar databases of different nationalities worldwide may expose ethnical influences on dental development. PMID- 19187461 TI - Operator exposure to scatter radiation from a portable hand-held dental radiation emitting device (Aribex NOMAD) while making 915 intraoral dental radiographs. AB - Operator exposure to backscatter radiation while using an Aribex NOMAD radiation emitting device (a portable, self-contained, cordless, hand-held dental X-ray unit) was determined while the operator employed various typical and atypical use scenarios during the exposure of 715 digital and/or film-based dental radiographs and 200 study control exposures. Study data was compared to the radiation safety occupational exposure annual Maximum Permissive Dose (MPD) of 50 mSv (5000 mrem) to determine the possible exposure risk to an unprotected operator using this device. The results showed the reproductive organs received the highest dose and the thyroid the least. The average operator whole body dose for the study was determined to be 0.047 mSv (4.47 mrem) or 0.09% of the annual MPD. Extrapolating the data as an expression of averaged annual operator exposure resulted in a whole body dose of 0.4536 mSv (45.36 mrem) or 0.9% of the annual MPD, These results are well below established occupation exposure limits and are compatible with those published by the manufacturer. PMID- 19187462 TI - Ca2+ channels and Ca2+-activated K+ channels in adult rat gonadotrophin-releasing hormone neurones. AB - Gonadotrophin-releasing hormone (GnRH) neurones represent the final output neurones in the neuroendocrine system for the control of reproduction. To understand the reproductive neuroendocrine system, an investigation of the intrinsic and extrinsic properties of GnRH neurones is essential. In this review, we focus on the intrinsic properties and summarise our recent findings of ion channels expressed in rat GnRH neurones. Rat GnRH neurones express all four types of high voltage-activated Ca(2+) channel (L, N, P/Q, R) and the low voltage activated Ca(2+) channel (T). GnRH neurones also express two types of Ca(2+) activated K(+) [K(Ca)] channel: the small conductance Ca(2+)-activated K(+) (SK) channel and the large conductance Ca(2+)- and voltage-activated K(+) (BK) channel. The activities of these Ca(2+) and K(Ca) channels regulate cell excitability and cellular calcium homeostasis. PMID- 19187464 TI - Gastrin-releasing peptide system in the spinal cord controls male sexual behaviour. AB - The lumbar spinal cord contains local neural circuits that are important in regulating male sexual behaviours, but the molecular mechanisms underlying these systems remain elusive. Gastrin-releasing peptide (GRP) is a member of the bombesin-like peptide family first isolated from the porcine stomach. Despite extensive pharmacological studies on the activity of bombesin-like peptides administered to mammals, little is known about the physiological functions of GRP in the spinal cord. We review recent findings on a system of neurones in the upper lumbar spinal cord, within the recently reported ejaculation generator, projecting axons containing GRP to the lower lumbar spinal cord and innervating regions known to control erection and ejaculation. PMID- 19187466 TI - Modulation of gonadotrophin-releasing hormone secretion by an endogenous circadian clock. AB - The mechanisms mediating positive feedback effects of oestradiol on pre-ovulatory gonadotrophin releasing-hormone (GnRH) surge generation in female mammals, although well-explored, are still incompletely understood. In addition to binding to and signalling through classical nuclear receptor-mediated pathways in afferent hypothalamic neurones, recent evidence suggests that ovarian steroids may use membrane-bound receptors or nonclassical signalling pathways to directly influence cell function leading to the generation of GnRH surge secretion. We review recent investigations into the role of the endogenous molecular circadian clock on modulation of GnRH gene expression and neuropeptide secretion, and will explore potential molecular mechanisms by which ovarian steroids may directly induce secretory changes at the level of the GnRH neurone, examining closely whether circadian clock gene oscillations may be involved. PMID- 19187465 TI - Cross-talk between membrane-initiated and nuclear-initiated oestrogen signalling in the hypothalamus. AB - It is increasingly evident that 17beta-oestradiol (E(2)), via a distinct membrane oestrogen receptor (Gq-mER), can rapidly activate kinase pathways to have multiple downstream actions in central nervous system (CNS) neurones. We have found that E(2) can rapidly reduce the potency of the GABA(B) receptor agonist baclofen and mu-opioid receptor agonist DAMGO to activate G-protein-coupled, inwardly rectifying K(+) (GIRK) channels in hypothalamic neurones, thereby increasing the excitability (firing activity) of pro-opiomelanocortin (POMC) and dopamine neurones. These effects are mimicked by the membrane impermeant E(2)-BSA and a new ligand (STX) that is selective for the Gq-mER that does not bind to ERalpha or ERbeta. Both E(2) and STX are fully efficacious in attenuating the GABA(B) response in ERalpha, ERbeta and GPR 30 knockout mice in an ICI 182 780 reversible manner. These findings are further proof that E(2) signals through a unique plasma membrane ER. We have characterised the coupling of this Gq-mER to a Gq-mediated activation of phospholipase C leading to the up-regulation of protein kinase Cdelta and protein kinase A activity in these neurones, which ultimately alters gene transcription. Finally, as proof of principle, we have found that STX, similar to E(2), reduces food intake and body weight gain in ovariectomised females. STX, presumably via the Gq-mER, also regulates gene expression of a number of relevant targets including cation channels and signalling molecules that are critical for regulating (as a prime example) POMC neuronal excitability. Therefore, E(2) can activate multiple receptor-mediated pathways to modulate excitability and gene transcription in CNS neurones that are critical for controlling homeostasis and motivated behaviors. PMID- 19187467 TI - Steroid hormone action in the brain: cross-talk between signalling pathways. AB - Ovarian steroid hormones, oestradiol and progesterone, modulate neuroendocrine functions in the central nervous system, resulting in alterations in physiology and behaviour. The classical model of steroid hormone action assumes that these neural effects are predominantly mediated via their intracellular receptors functioning as 'ligand-dependent' transcription factors in the steroid-sensitive neurones regulating genes and genomic networks with profound behavioural consequences. Studies from our laboratory demonstrate that, in addition to their cognate ligands, intracellular steroid receptors can be activated in a 'ligand independent' manner by the neurotransmitter dopamine, which alters the dynamic equilibrium between neuronal phosphatases and kinases. A high degree of cross talk between membrane-initiated signalling pathways and the classical intracellular signalling pathways mediates hormone-dependent behaviour in mammals. The molecular mechanisms, by which a multitude of signals converge with steroid receptors to delineate a genomic level of cross-talk in brain and behaviour are discussed. PMID- 19187468 TI - Examining the intersection of sex and stress in modelling neuropsychiatric disorders. AB - Sex-biased neuropsychiatric disorders, including major depressive disorder and schizophrenia, are the major cause of disability in the developed world. Elevated stress sensitivity has been proposed as a key underlying factor in disease onset. Sex differences in stress sensitivity are associated with corticotrophin releasing factor (CRF) and serotonin neurotransmission, which are important central regulators of mood and coping responses. To elucidate the underlying neurobiology of stress-related disease predisposition, it is critical to develop appropriate animal models of stress pathway dysregulation. Furthermore, the inclusion of sex difference comparisons in stress responsive behaviours, physiology and central stress pathway maturation in these models is essential. Recent studies by our laboratory and others have begun to investigate the intersection of stress and sex where the development of mouse models of stress pathway dysregulation via prenatal stress experience or early-life manipulations has provided insight into points of developmental vulnerability. In addition, examination of the maturation of these pathways, including the functional importance of the organisational and activational effects of gonadal hormones on stress responsivity, is essential for determination of when sex differences in stress sensitivity may begin. In such studies, we have detected distinct sex differences in stress coping strategies where activational effects of testosterone produced females that displayed male-like strategies in tests of passive coping, but were similar to females in tests of active coping. In a second model of elevated stress sensitivity, male mice experiencing prenatal stress early in gestation showed feminised physiological and behavioural stress responses, and were highly sensitive to a low dose of selective serotonin reuptake inhibitors. Analyses of expression and epigenetic patterns revealed changes in CRF and glucocorticoid receptor genes in these mice. Mechanistically, stress early in pregnancy produced a significant sex-dependent effect on placental gene expression that was supportive of altered foetal transport of key growth factors and nutrients. These mouse models examining alterations and hormonal effects on development of stress pathways provide necessary insight into how specific stress responses can be reprogrammed early in development resulting in sex differences in stress sensitivity and neuropsychiatric disease vulnerability. PMID- 19187470 TI - Parasympathetic nervous system dysfunction and neurological consequences of chagas disease. Mediator or bystander? PMID- 19187471 TI - Uncovering and characterizing multiple sclerosis lesions: the aid of fluid attenuated inversion recovery images in the presence of gadolinium contrast agent. PMID- 19187472 TI - Neuroplastic changes in the brain: a case of two successive adaptive changes within the motor cortex. AB - We describe a case of neuroplasticity associated with both arteriovenous malformation (AVM) and stroke, which occurred in two successive events in the same patient. Functional magnetic resonance imaging (fMRI) during right-hand movement in a young man with a left rolandic AVM detected activation of a region corresponding to the left premotor cortex. The AVM was embolized. A few hours after the last embolization session, the patient sustained an ischemic complication in the left subcortical white matter. A second fMRI detected a lower degree of left premotor cortex activation and strong activation of the contralesional right primary motor cortex and bilateral supplementary motor areas. One month later, in association with clinical recovery, the fMRI activation returned to that observed in the first fMRI, ie, selective activation of the ipsilesional left premotor cortex. This is, to our knowledge, the first description of two distinct functional cortical changes determined by an AVM and a stroke within the motor network. PMID- 19187473 TI - MR and MRS characteristics of intraventricular meningioma. AB - Meningiomas are frequent intracranial, non-glial tumors of adults. We present the unusual left lateral ventricular localization of meningioma in a 51-year-old man. The magnetic resonance (MR) images showed well demarcated, large mass of the atrium of the left lateral ventricle with transependymal extension into the left temporal lobe. MR spectroscopy revealed the presence of "choline only" spectrum, typical for extra axial neoplasms. The mass was completely resected. The diagnosis of transitional type intraventricular meningioma, with psammoma bodies, histologic grade I was made. Progesterone and estrogen receptors were negative. PMID- 19187469 TI - Effects of gonadotrophin-releasing hormone outside the hypothalamic-pituitary reproductive axis. AB - Gonadotrophin-releasing hormone (GnRH) is a hypothalamic decapeptide with an undisputed role as a primary regulator of gonadal function. It exerts this regulation by controlling the release of gonadotrophins. However, it is becoming apparent that GnRH may have a variety of other vital roles in normal physiology. A reconsideration of the potential widespread action that this traditional reproductive hormone exerts may lead to the generation of novel therapies and provide insight into seemingly incongruent outcomes from current treatments using GnRH analogues to combat diseases such as prostate cancer. PMID- 19187474 TI - Clinical and MRI characteristics of levamisole-induced leukoencephalopathy in 16 patients. AB - OBJECTIVE: To study the clinical and radiological characteristics of levamisole induced leukoencephalopathy (LILE) in patients with recurrent aphthous ulcers (RAU) or infected with Ascaris. METHODS: The medical histories of 16 patients with LILE were analyzed, including clinical manifestations, brain magnetic resonance imaging (MRI) characteristics, cerebrospinal fluid, and brain biopsy findings. RESULTS: The main clinical manifestations of LILE were motor weakness (75.0%), dysphasia or aphasia (50.0%), cognitive disorder (50.0%), and facial palsy (43.8%). The MRI of 16 cases showed plaque and round or oval demyelinating lesions in white matter, which revealed a signal hypointensity on T1-weighted and diffusion-weighted images (DWI), and demonstrated hyperintensity on T2-weighted and fluid-attenuated inversion recovery (FLAIR) images. The MRI revealed peripheral ring-type enhancement about the focus after Gd-DTPA administration and edema around some lesions, without mass effect. T2-weighted and FLAIR images were highly sensitive to the lesions. Brain biopsy in 1 patient showed multifocal demyelinating lesions without perivascular cuffing by lymphocytes. Treatment generally consisted of steroids and hyperbaric oxygen therapy, and patients recovered to normal condition. CONCLUSIONS: A single normal dose of levamisole can induce leukoencephalopathy in patients with RAU or Ascaris. The diagnosis depends on the clinical features and imaging appearances. Steroid therapy might be a good choice of treatment. PMID- 19187475 TI - Voxel-based morphometry and Voxel-based relaxometry in parkinsonian variant of multiple system atrophy. AB - BACKGROUND AND PURPOSE: Multiple system atrophy (MSA) is a progressive neurodegenerative disorder divided into a parkinsonian (MSA-P) and a cerebellar variant. The purpose of this study was to assess regional brain atrophy and iron content using Voxel-based morphometry (VBM) and Voxel-based relaxometry (VBR) respectively, in MSA-P. METHODS: Using biological parametric mapping the effect of brain atrophy was evaluated in T2 relaxation time (T2) measurements by applying analysis of covariance (ANCOVA) and correlation analysis to the VBM and VBR data. Eleven patients with MSA-P (aged 61.9 +/- 11.7 years, disease duration 5.42 +/- 2.5 years) and 11 controls were studied. RESULTS: In comparison to the controls the patients showed decreased gray matter in the putamen, the caudate nuclei, the thalami, the anterior cerebellar lobes, and the cerebral cortex, and white matter atrophy in the pons, midbrain, and peduncles. VBR analysis showed prolonged T2 in various cortical regions. On ANCOVA, when controlling for gray and white matter volume, these regions of prolonged T2 were shrunk. Negative correlation was demonstrated between T2 and gray and white matter volume. CONCLUSIONS: Diffuse brain atrophy, mainly in the motor circuitry is observed in MSA-P. Normalization for atrophy should always be performed in T2 measurements. PMID- 19187476 TI - Seizure-induced inflammation in focal cortical dysplasia resulting in imaging progression that simulates neoplasia. AB - A 5-year-old girl with previously well-controlled partial epilepsy secondary to focal cortical dysplasia (FCD) developed an increase in seizure frequency. Two months later, magnetic resonance showed a substantial alteration in lesion imaging characteristics. The lesion was resected. FCD was confirmed but inflammatory changes were also present. We propose that chronic inflammation was induced by unremitting seizure activity and suggest that inflammation may be implicated as a basis for alteration in the imaging characteristics of FCD. PMID- 19187477 TI - Temporary test occlusion of distal middle cerebral artery using GDC coil. AB - The treatment of small distal intracranial aneurysms often requires parent vessel occlusion. However, this may be particularly challenging in patients in which the parent vessel serves eloquent brain. Superselective amytal testing is often conducted in these cases, but may prove unreliable or inconclusive. In order to more specifically assess the functional significance of the parent vessel in this patient with a distal left middle cerebral artery pseudoaneurysm who had failed superselective amytal testing, we used a Guglielmi Detachable coil to perform a temporary test occlusion. Testing was performed with the patient awake and examinable, and after no neurological changes were noted, the vessel was permanently occluded, successfully treating the aneurysm. To our knowledge, this is the first report to describe this unique method for assessing a parent vessel that previously had been thought to supply eloquent brain during amytal testing. PMID- 19187479 TI - Pilot study of new diagnostic criteria for middle cerebral artery stenosis by transcranial Doppler. AB - BACKGROUND AND PURPOSE: Current criteria for middle cerebral artery (MCA) stenosis by transcranial Doppler were mainly based on the flow velocity acceleration, ignoring other useful parameters. We tested the accuracy of new criteria using magnetic resonance angiography (MRA) as the confirmative method. METHODS: A compositive score for each MCA was calculated according to following parameters: Velocity Scale (score 0-6 for peak systolic velocity <140 to > or = 300 cm/second), Hemodynamic Scale (score 0-5 for diffuse or focal velocity increase; score 0-6 for differences between bilateral MCA; score 17 for damping velocity), and Spectrum Scale (score 0-2 for normal spectrum, turbulence, and musical murmurs). RESULTS: The median scores in normal-mild, moderate, and severe groups on MRA differed significantly (P < .001). The optimal cutoff score for <50% and > or = 50% stenosis was 3 with the PPV of 72.3% (95% confidence interval [CI]: 65.3% to 78.5%) and negative predictive value (NPV) of 89.5% (95% CI: 85.9% to 92.3%), for moderate and severe stenosis was 6 with the PPV of 85.9% (95% CI: 76.7% to 92.0%) and NPV of 61.9% (95% CI: 50.6% to 72.1%). Both scores performed better than peak systolic or mean velocity. CONCLUSIONS: This new method was feasible and reliable and showed some advantages over the velocity-only method. Further studies using digital subtraction angiography as the gold standard are needed to ameliorate the new criteria. PMID- 19187478 TI - High-resolution phased-array MRI of the human brain at 7 tesla: initial experience in multiple sclerosis patients. AB - Recent advancement for magnetic resonance imaging (MRI) involves the incorporation of higher-field strengths. Although imagers with higher magnetic field strengths were developed and tested in research labs, the direct application to patient MR studies have been extremely limited. Imaging at 7 Tesla (7T) affords advantages in signal-to-noise ratio and image contrast and resolution; however, these benefits can only be realized if the correct coils exist to capture the images. The objective of this study was to develop optimized high-resolution 7T MRI techniques using high sensitivity, specialized phased array coils, for improved gray matter (GM) and white matter differentiation, in an effort to improve visualization of multiple sclerosis (MS) lesions in vivo. Twenty-three subjects were enrolled in this preliminary study, 17 with clinically definite MS (11 females, 6 males; mean age 43.4 years; range 22-64 years) and 6 healthy controls (2 females, 4 males; mean age 39.0 years; range 27-67 years). MR imaging of MS patients at 7T was demonstrated to be safe, well tolerated, and provided high-resolution anatomical images allowing visualization of structural abnormalities localized near or within the cortical layers. Clear involvement of the GM was observed with improved morphological detail in comparison to imaging at lower-field strength. PMID- 19187480 TI - MRI features of pontine autosomal dominant microangiopathy and leukoencephalopathy (PADMAL). AB - BACKGROUND AND PURPOSE: Cerebral small vessel disease is a topic of growing interest for both the scientific community and the aging society. We report the magnetic resonance imaging (MRI) characteristics of a recently found autosomal dominantly inherited microangiopathy. METHODS: Eighteen members (35 to 77 years) of a large German family underwent MR scanning with a standardized MRI protocol for cerebrovascular diseases. Images were evaluated independently by two neuroradiologists. RESULTS: Six family members revealed an unequivocally pathological phenotype on MRI with lacunar infarcts of the pons (6/6) and lesions of the subcortical and periventricular white matter (5/6). Lesions in the temporal lobes (1/6) and cerebral microbleeds (1/6) were uncommon. None of the patients revealed atherosclerotic changes in MR angiography. Retrospective analysis of 5 brain autopsies from previously reported patients of the same family confirmed the regular involvement of the pons. CONCLUSION: This cerebral autosomal dominant arteriopathy with pontine infarcts and leukoencephalopathy is characterized by a special lesion pattern strikingly different from CADASIL. The distinct MRI characteristics with pontine lesions and rare occurrence of temporal lesions argue for a new nosological entity and may be helpful for the differential diagnosis. PMID- 19187481 TI - Decreased cortical somatosensory finger representation in X-linked recessive bulbospinal neuronopathy (Kennedy disease): a magnetoencephalographic study. AB - BACKGROUND: Kennedy disease (KD) clinically presents as progressive lower motor neuron disease with minimal or no sensory impairment. However, electrophysiological studies found abnormal somatosensory-evoked potentials even in absence of clinical deficits. Little is known about possible influences of this sensory neuropathy on the central somatosensory processing. METHODS: In this study, the cortical topography of index finger representation was studied in 7 patients with genetically proven KD compared to healthy control subjects by means of magnetoencephalography using an established stimulation paradigm. Data analysis was carried out with synthetic aperture magnetometry (SAM). Additionally, the latency and source amplitude of the earliest cortical somatosensory-evoked field (SEF) component were determined based on traditional single dipole source analysis. RESULTS: In KD patients the latency of the SEF was prolonged (48.6 vs. 37.4 ms, P < .005). There was no significant difference in dipole source amplitude, but stimulus-related SAM activation of the contralateral sensorimotor cortex (pseudo-t-values -.107 vs. -.199, P < .05), including maximum activity (53.5%), was reduced. CONCLUSIONS: These results implicate that even subclinical sensory neuropathy leads to possible functional reorganization of the sensorimotor cortex in KD patients and reinforces the view that in KD the somatosensory system is extensively involved. PMID- 19187482 TI - A new theoretical discrete growth distribution with verification for microbial counts in water. AB - Living microbes are discrete, not homogeneously distributed in environmental media, and the form of the distribution of their counts in drinking water has not been well established. However, this count may "scale" or range over orders of magnitude over time, in which case data representing the tail of the distribution, and governing the mean, would be represented only in impractically long data records. In the absence of such data, knowledge of the general form of the full distribution could be used to estimate the true mean accounting for low probability, high-consequence count events and provide a basis for a general environmental dose-response function. In this article, a new theoretical discrete growth distribution (DGD) is proposed for discrete counts in environmental media and other discrete growth systems. The term growth refers not to microbial growth but to a general abiotic first-order growth/decay of outcome sizes in many complex systems. The emergence and stability of the DGD in such systems, defined in simultaneous work, are also described. The DGD is then initially verified versus 12 of 12 simulated long-term drinking water and short-term treated and untreated water microbial count data sets. The alternative Poisson lognormal (PLN) distribution was rejected for 2 (17%) of the 12 data sets with 95% confidence and, like other competitive distributions, was not found stable (in simultaneous work). Sample averages are compared with means assessed from the fitted DGD, with varying results. Broader validation of the DGD for discrete counts arising as outcomes of mathematical growth systems is suggested. PMID- 19187483 TI - Predicting future excess events in risk assessment. AB - Risk characterization in a study population relies on cases of disease or death that are causally related to the exposure under study. The number of such cases, so-called "excess" cases, is not just an indicator of the impact of the risk factor in the study population, but also an important determinant of statistical power for assessing aspects of risk such as age-time trends and susceptible subgroups. In determining how large a population to study and/or how long to follow a study population to accumulate sufficient excess cases, it is necessary to predict future risk. In this study, focusing on models involving excess risk with possible effect modification, we describe a method for predicting the expected magnitude of numbers of excess cases and assess the uncertainty in those predictions. We do this by extending Bayesian APC models for rate projection to include exposure-related excess risk with possible effect modification by, e.g., age at exposure and attained age. The method is illustrated using the follow-up study of Japanese Atomic-Bomb Survivors, one of the primary bases for determining long-term health effects of radiation exposure and assessment of risk for radiation protection purposes. Using models selected by a predictive-performance measure obtained on test data reserved for cross-validation, we project excess counts due to radiation exposure and lifetime risk measures (risk of exposure induced deaths (REID) and loss of life expectancy (LLE)) associated with cancer and noncancer disease deaths in the A-Bomb survivor cohort. PMID- 19187484 TI - A model of exposure to rotavirus from nondietary ingestion iterated by simulated intermittent contacts. AB - Existing microbial risk assessment models rarely incorporate detailed descriptions of human interaction with fomites. We develop a stochastic mechanistic model of exposure to rotavirus from nondietary ingestion iterated by simulated intermittent fomes-mouth, hand-mouth, and hand-fomes contacts typical of a child under six years of age. This exposure is subsequently translated to risk using a simple static dose-response relationship. Through laboratory experiments, we quantified the mean rate of inactivation for MS2 phage on glass (0.0052/hr) and mean transfer between fingertips and glass (36%). Simulations using these parameters demonstrated that a child's ingested dose from a rotavirus contaminated ball ranges from 2 to 1,000 virus over a period of one hour, with a median value of 42 virus. These results were heavily influenced by selected values of model parameters, most notably the concentration of rotavirus on fomes, frequency of fomes-mouth contacts, frequency of hand-mouth contacts, and virus transferred from fomes to mouth. The model demonstrated that mouthing of fomes is the primary exposure route, with hand mouthing contributions accounting for less than one-fifth of the child's dose over the first 10 minutes of interaction. PMID- 19187485 TI - Loss of life caused by the flooding of New Orleans after Hurricane Katrina: analysis of the relationship between flood characteristics and mortality. AB - In this article a preliminary analysis of the loss of life caused by Hurricane Katrina in the New Orleans metropolitan area is presented. The hurricane caused more than 1,100 fatalities in the state of Louisiana. A preliminary data set that gives information on the recovery locations and individual characteristics for 771 fatalities has been analyzed. One-third of the analyzed fatalities occurred outside the flooded areas or in hospitals and shelters in the flooded area. These fatalities were due to the adverse public health situation that developed after the floods. Two-thirds of the analyzed fatalities were most likely associated with the direct physical impacts of the flood and mostly caused by drowning. The majority of victims were elderly: nearly 60% of fatalities were over 65 years old. Similar to historical flood events, mortality rates were highest in areas near severe breaches and in areas with large water depths. An empirical relationship has been derived between the water depth and mortality and this has been compared with similar mortality functions proposed based on data for other flood events. The overall mortality among the exposed population for this event was approximately 1%, which is similar to findings for historical flood events. Despite the fact that the presented results are preliminary they give important insights into the determinants of loss of life and the relationship between mortality and flood characteristics. PMID- 19187486 TI - Risk-based decision making for terrorism applications. AB - This article describes the anti-terrorism risk-based decision aid (ARDA), a risk based decision-making approach for prioritizing anti-terrorism measures. The ARDA model was developed as part of a larger effort to assess investments for protecting U.S. Navy assets at risk and determine whether the most effective anti terrorism alternatives are being used to reduce the risk to the facilities and war-fighting assets. With ARDA and some support from subject matter experts, we examine thousands of scenarios composed of 15 attack modes against 160 facility types on two installations and hundreds of portfolios of 22 mitigation alternatives. ARDA uses multiattribute utility theory to solve some of the commonly identified challenges in security risk analysis. This article describes the process and documents lessons learned from applying the ARDA model for this application. PMID- 19187487 TI - Time-dose-response models for microbial risk assessment. AB - While microbial risk assessment (MRA) has been used for over 25 years, traditional dose-response analysis has only predicted the overall risk of adverse consequences from exposure to a given dose. An important issue for consequence assessment from bioterrorist and other microbiological exposure is the distribution of cases over time due to the initial exposure. In this study, the classical exponential and beta-Poisson dose-response models were modified to include exponential-power dependency of time post inoculation (TPI) or its simplified form, exponential-reciprocal dependency of TPI, to quantify the time of onset of an effect presumably associated with the kinetics of in vivo bacterial growth. Using the maximum likelihood estimation approach, the resulting time-dose-response models were found capable of providing statistically acceptable fits to all tested pooled animal survival dose-response data. These new models can consequently describe the development of animal infectious response over time and represent observed responses fairly accurately. This is the first study showing that a time-dose-response model can be developed for describing infections initiated by various pathogens. It provides an advanced approach for future MRA frameworks. PMID- 19187488 TI - Lactoferrin and desferrioxamine are ineffective in the treatment of Helicobacter pylori infection and may enhance H. pylori growth and gastric inflammation in mice. AB - AIMS: To evaluate the efficacy of bovine lactoferrin (BLf), recombinant human lactoferrin (rHLf) and desferrioxamine against Helicobacter pylori in vitro and in mice and also to determine whether BLf or rHLf alter gastric inflammation. METHODS AND RESULTS: In vitro: Broth dilution susceptibility tests were performed using different concentrations of desferrioxamine, BLf and rHLf. Murine trials: In the prevention trial, C57BL/6 female mice were treated with BLf or rHLF, and then infected with the SS1 strain of H. pylori. In the treatment trial, mice were gavaged with either BLf, rHLf or desferrioxamine. In addition, gastric myeloperoxidase activity (MPO) was measured to assess gastric inflammation. Desferoxamine was found to have a direct bactericidal effect, while BLf and rHLf only partially suppressed H. pylori growth in vitro. However, in both prevention and treatment trials all three forms of treatment failed to reduce H. pylori load in mice. Gastric MPO activity and H. pylori load were noted to be higher with lactoferrin treatments. CONCLUSIONS: Our study does not support the use of BLf or rHLF in the treatment of human H. pylori infection. Interestingly, H. pylori growth and gastric inflammation appear to be enhanced by lactoferrin treatment. SIGNIFICANCE AND IMPACT OF THE STUDY: The mouse model is ideal for testing novel H. pylori eradicating agents. PMID- 19187489 TI - Study of the coumarate decarboxylase and vinylphenol reductase activities of Dekkera bruxellensis (anamorph Brettanomyces bruxellensis) isolates. AB - AIM: To evaluate the coumarate descarboxylase (CD) and vinylphenol reductase (VR) activities in Dekkera bruxellensis isolates and study their relationship to the growth rate, protein profile and random amplified polymorphic DNA (RAPD) molecular pattern. METHODS AND RESULTS: CD and VR activities were quantified, as well, the growth rate, intracellular protein profile and molecular analysis (RAPD) were determined in 12 isolates of D. bruxellensis. All the isolates studied showed CD activity, but only some showed VR activity. Those isolates with the greatest growth rate did not present a different protein profile from the others. The FASC showed a relationship between RAPD molecular patterns and VR activity. CONCLUSION: CD activity is common to all of the D. bruxellensis isolates. This was not the case with VR activity, which was detected at a low percentage in the analysed micro-organisms. A correlation was observed between VR activity and the RAPD patterns. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study that quantifies the CD and VR enzyme activities in D. bruxellensis, demonstrating that these activities are not present in all isolates of this yeast. PMID- 19187490 TI - Nonpathogenic Entamoeba dispar quickly outgrows pathogenic Entamoeba histolytica in mixed xenic cultures. AB - Entamoeba histolytica and Entamoeba dispar are two microscopically indistinguishable amoebae living in the human colon. The former is a pathogen, whereas the latter is a nonpathogenic commensal. Using a model system of in vitro cocultures and PCR detection of the Entamoeba species, we found that the nonpathogenic species can rapidly outgrow the pathogen in xenic cultures. PMID- 19187491 TI - Analyses of microbial consortia in the starter of Fen Liquor. AB - AIMS: This study aimed to determine the microbial diversity in the starter of Fen Liquor. METHODS AND RESULTS: The plate method was used to enumerate the micro organisms; meanwhile, the 16S rDNA of bacteria and the internal transcribed spacer of fungi were used to determine microbial diversity. Several genera were accordingly identified. Among the bacteria, Lactobacillales and Actinomycetales were detected only on the surface of the starter, whereas Bacillales was dominant within the starter. Among the fungi, Saccharomycopsis and Issatchenkia were the main genera in surface and interior starter, respectively; in addition, Thermomyces was found in interior starter, while other species of fungi were detected on the surface. CONCLUSIONS: The culture-dependent and polymerase chain reaction-based methods revealed the significant microbial diversity in different locations in the starter of Fen Liquor. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is the first to identify the bacterial and fungal communities associated with the starter of Fen Liquor using both traditional and molecular methods; it is also the first to compare the microbial diversity on the surface of starter with that in the interior. The results enrich our knowledge on liquor related micro-organisms, and can be used to promote the development of the traditional fermentation technology. PMID- 19187492 TI - Prevalence of Helicobacter and Acanthamoeba in natural environment. AB - AIMS: We examined whether the presence of Helicobacter is related to that of Acanthamoeba in river and soil environments. METHODS AND RESULTS: The samples (river n = 51, soil n = 75) were collected in Sapporo City, Japan. PCR with primers for Helicobacter genus-specific and standard culture techniques were used to detect helicobacter. Prevalence of acanthamoeba was also evaluated by genus specific PCR. The prevalence of Helicobacter genus-specific DNA in river water samples and in soil samples was 88% and 0%, respectively. No successful culture of helicobacter was achieved. The prevalence of Acanthamoeba genus-specific DNA in river samples and in soil samples was 61% and 96%, respectively. No statistical correlation between the prevalence of helicobacter and either that of acanthamoeba or water quality parameters (pH, turbidity and coliform group) except for temperature was found. CONCLUSIONS: We revealed the presence of helicobacter in river water and non-existence of helicobacter in soil. However, the distribution of helicobacter did not overlap with that of acanthamoeba in rivers. SIGNIFICANCE FOR IMPACT OF THE STUDY: The role of acanthamoeba on the survival of helicobacter might be limited as the both are coincidentally present in the environment. PMID- 19187493 TI - Screening of food raw materials for the presence of Shiga toxin-producing Escherichia coli O91:H21. AB - AIMS: To provide information on the prevalence and detection, in foods, of Shiga toxin-producing Escherichia coli (STEC) O91:H21. METHODS AND RESULTS: Seven hundred fifteen minced beef meats and 205 raw milk samples were analysed by stx specific PCR-ELISA. Samples positive for stx were subsequently tested for the presence of wzy-O91, fliC-H21 and the adhesin-encoding gene saa. For minced meat, 16 (2.2%) and 11 (1.5%) samples were found positive for (stx, wzy-O91, fliC-H21) and (stx, wzy-O91, fliC-H21, saa) combinations, respectively. For raw milk, seven (3.4%) samples were found positive for the (stx, wzy-O91, fliC-H21) combination but none of these contained saa. Two STEC O91:H21 saa-positive strains and three STEC O91 H21- and saa-negative strains were isolated by colony hybridization. CONCLUSIONS: A low prevalence of potentially pathogenic STEC O91:H21 in food products was found using a combination of PCR assays targeting stx, wzy-O91, fliC H21 and saa. SIGNIFICANCE AND IMPACT OF THE STUDY: The PCR-based approach described here represents a valuable method for rapid screening of food samples contaminated by STEC O91:H21. PMID- 19187494 TI - The antimicrobial activity of four commercial essential oils in combination with conventional antimicrobials. AB - AIMS: Due to the emergence of multi-drug resistance, alternatives to conventional antimicrobial therapy are needed. This study aims to investigate the in vitro pharmacological interactions between essential oils (considered valuable as natural therapeutic treatments) and conventional antimicrobials (ciprofloxacin/amphotericin B) when used in combination. METHODS AND RESULTS: Interactions of the essential oils (Melaleuca alternifolia, Thymus vulgaris, Mentha piperita and Rosmarinus officinalis) when combined with ciprofloxacin against Staphylococcus aureus indicate mainly antagonistic profiles. When tested against Klebsiella pneumoniae the isobolograms show antagonistic, synergistic and additive interactions depending on the combined ratio. The R. officinalis/ciprofloxacin combination against K. pneumoniae displayed the most favourable synergistic pattern. The interactions of M. alternifolia (tea tree), T. vulgaris (thyme), M. piperita (peppermint) and R. officinalis (rosemary) essential oils with amphotericin B indicate mainly antagonistic profiles when tested against Candida albicans. CONCLUSION: While a number of interactions show complete antagonism, others show varied (synergistic, additive and/or antagonistic) interactions, thus the efficacy is dependent on the ratio in which the two components co-exist. SIGNIFICANCE AND IMPACT OF THE STUDY: The predominant antagonistic interactions noted here, suggests that some natural therapies containing essential oils should be used with caution when combined with antibiotics. PMID- 19187495 TI - Novel 16S rRNA gene analyses reveal new in vitro effects of insoluble barley fibres on the human faecal microbiota. AB - AIMS: The aim of this work was to analyse the growth of human faecal microbiota on barley dietary fibres (DF). It is generally accepted that insoluble DF are health promoting, but the information is scarce about how these fibres affect the gastrointestinal (GI) microbiota. A major reason for the limited knowledge is that there are currently no proper tools to analyse the complete GI microbiota. METHODS AND RESULTS: Here we present a novel 16S rRNA gene analytical approach that enables the analyses of the complete microbiota, including the part that has not yet been characterized. The basic principle of the method is use of 16S rRNA gene signature sequences to determine both the phylogenetic relatedness and the distribution of bacteria in the samples analysed. Using this approach, we analysed the microbiota after in vitro fermentation of different barley fractions with human faeces. Our main finding was that groups of actinobacteria were selectively enriched by growth on the insoluble DF fractions. CONCLUSIONS: Our novel analytical approaches revealed new enrichment patterns in the taxa that respond to insoluble DF. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results may have major implications for future understanding of insoluble DF health effects. PMID- 19187496 TI - Development of a defined medium for Clostridium scatologenes ATCC 25775. AB - AIMS: To develop a defined medium for Clostridium scatologenes ATCC 25775, which produces the malodorants 3-methylindole (skatole) and 4-methylphenol (p-cresol). METHODS AND RESULTS: Clostridium scatologenes was cultured in anaerobic broth medium (pH 6.3) at 37 degrees C containing ammonia, minerals and a commercial vitamin solution. Data indicate alpha-ketoglutarate, L-glutamate or L-glutamine is a required nutrient that can also serve as a primary carbon and energy source. When cultured in defined medium containing glutamate; glucose, fructose and betaine served as primary carbon and energy sources. L-Tryptophan, L-tyrosine, sorbitol and indole acetic acid did not enhance growth. In the absence of tryptophan, cells produced indole when grown using glucose or fructose. 4 Methylphenol was produced when growing cells were supplied with tyrosine. When supplied with tryptophan, 3-methylindole was produced by glucose- or fructose growing cells but not from glutamate-growing cells. Cells grown in the presence of pyruvate produced indole, 3-methylindole and 4-methylphenol. CONCLUSIONS: Clostridium scatologenes requires alpha-ketoglutarate, L-glutamate, or L glutamine for growth in defined medium. Cells produce indole when glucose or fructose is included in defined medium. SIGNIFICANCE AND IMPACT OF THE STUDY: The development of a defined medium will assist in physiology studies and genetic analysis of this strain. PMID- 19187497 TI - Effect of physical conditions and chemicals on the binding of a mini-CbpA from Clostridium cellulovorans to a semi-crystalline cellulose ligand. AB - AIMS: To investigate the effect that environmental factors have on Clostridium cellulovorans cellulose binding domain (CBD) binding to a semi-crystalline cellulose ligand, namely Avicel. METHODS AND RESULTS: The behaviour of a 58 kDa mini-CbpA protein containing the CBD from the scaffoldin protein of C. cellulovorans was studied in the presence of various environmental factors, in order to determine whether such factors promote or reduce CBD binding to its ligand, thus potentially affecting its activity on the substrate. The amount of binding was found to be dependent on the Avicel concentration and optimal binding occurred when the ligand concentration was 15 mg ml(-1). Optimal CBD binding occurred at pH 7.0 and at an incubation temperature of 28 degrees C. The effects of dithiothreitol (DTT), 2-mercaptoethanol, acetone, butanol, ethanol and butyric acid were also investigated. CONCLUSIONS: Temperature, pH, DTT, 2-mercaptoethanol and solvents were shown to affect the binding of C. cellulovorans CBD to Avicel. SIGNIFICANCE AND IMPACT OF THE STUDY: Clostridium cellulovorans CBD binding to Avicel is affected by physical conditions and chemicals. PMID- 19187498 TI - An image analysis technique to estimate the cell density and biomass concentration of Trichoderma reesei. AB - AIM: The objective is to develop an automated image analysis protocol to quantify the cell volume fraction of filamentous fungi (Trichoderma reesei) and estimate the biomass concentration. METHODS AND RESULTS: Both dry weight and image analyses were performed on samples collected periodically from 7-l stirred tank fermentations. Using the projected area of lactophenol blue-stained hyphae, the fraction occupied by the cells in a given volume was estimated. Combined with the biomass dry weight obtained by filtration, the method was used to estimate the density of filamentous fungi. Knowing the density of fungi, the algorithm was employed to quantitatively assess the biomass evolution during the course of fermentation even in the presence of solid particles. CONCLUSIONS: A density of 0.334 g dry weight cm(-3) was found for T. reesei RUT C-30. The image analysis protocol allowed successful estimation of biomass concentration in the presence or absence of solid particles. SIGNIFICANCE AND IMPACT OF THE STUDY: Methods to quantify biomass during the industrial production of cellulase with T. reesei are often limited due to the presence of solid substrates. The image analysis protocol presented here offers a quick and easy way to estimate biomass concentration of filamentous micro-organisms in insoluble medium. PMID- 19187499 TI - Differential cell surface properties of vegetative Bacillus. AB - AIMS: The genus Bacillus encompasses a wide range of species which display varying pathogenic abilities. The hydrophobicity of a range of Bacillus species was determined to evaluate the correlation between bacterial hydrophobicity and pathogenicity. METHODS AND RESULTS: Bacterial adhesion to hydrocarbon assays were used to determine the hydrophobicity of various Bacillus species. Significant differences in the hydrophobicity of vegetative Bacilli were found. Specifically, vegetative Bacillus anthracis or Bacillus thuringiensis cells were highly hydrophobic whereas Bacillus cereus or Bacillus subtilis were only slightly hydrophobic using this test. Cell adhesion assays using A549 or J774 cells were used to demonstrate a correlation between the bacterial hydrophobicity profiles with the ability to adhere to the mammalian cell lines. CONCLUSIONS: The ability of Bacillus species to adhere to mammalian cell lines correlates with the hydrophobicity of the bacteria and also correlates with the relative pathogenicity of some of the Bacillus species tested. SIGNIFICANCE AND IMPACT OF THE STUDY: This work suggests that study of the physical-chemical properties of vegetative cells could inform future approaches for the rapid identification and discrimination of potentially pathogenic Bacilli. PMID- 19187500 TI - A combined immunomagnetic separation and lateral flow method for a sensitive on site detection of Bacillus anthracis spores--assessment in water and dairy products. AB - AIM: Combination of immunomagnetic separation (IMS) and lateral flow device (LFD) assays for the development of a sensitive, rapid, on-site methodology that enables concentration and detection of Bacillus anthracis spores in complex samples. METHODS AND RESULTS: The data presents the development of an optimized, 30 min, IMS assay, with about 95% capture of B. anthracis spores from different dairy products (n = 38). No cross reactivity was detected with typical milk flora and some closely related Bacilli. To enable direct application of the IMS captured spores on the LFD, spores were eluted from the bead-spore complex utilizing 95% (v/v) formamide-10 mmol l(-1) EDTA for 30 s in a microwave oven. Detached spores were analysed on LFD enabling detection within 10 min. The combined IMS-LFD methodology (40 min) demonstrates a 60-fold improvement in sensitivity, relative to samples that were applied directly on the LFD without the IMS concentrating step. CONCLUSIONS: The IMS-LFD method is a powerful platform, combining rapidity, specificity and efficiency for concentrating and detecting B. anthracis from water and milk contaminated samples. SIGNIFICANT AND IMPACT OF THE STUDY: The combination of IMS and LFD enhances the sensitivity and flexibility of B. anthracis spore detection from complex samples. This method can potentially be extended to other toxins and micro-organisms in a variety of matrices. PMID- 19187501 TI - Campylobacters and bacteriophages in the surface waters of Canterbury (New Zealand). AB - AIM: To determine the relationship between the presence of thermotolerant campylobacters and their bacteriophages (phages) in surface waters for the potential to use phages as an indicator of Campylobacter spp. METHODS AND RESULTS: Thermotolerant campylobacters were enumerated in 53 water samples using a three tube most probable number (MPN) series in m-Exeter broth. The presence of phages in the same samples was tested using two approaches: qualitative enrichment with five different Campylobacter hosts and a quantitative membrane concentration method. Phages infecting an Escherichia coli O157:H7 isolate were also enumerated by the membrane concentration method. Campylobacter spp. were isolated from 45/53 (85%) of the samples at 0.4-110 MPN 100 ml(-1). No Campylobacter phages were isolated, but coliphages were present in 43/46 (93%) of samples. CONCLUSIONS: The membrane concentration method recovered >80% of Campylobacter phages from spiked samples. The absence of Campylobacter phages in environmental samples, from both enrichment and concentration methods, suggests that, if present, they are at very low titres. SIGNIFICANCE AND IMPACT OF THE STUDY: Testing for Campylobacter phages as an indicator of Campylobacter spp. presence is not effective. The quantitative data for Campylobacter spp. will be useful for risk assessment purposes. PMID- 19187502 TI - Evaluation of the effectiveness of commercially available contact plates for monitoring microbial environments. AB - AIM: The aim of this study was to measure the efficiency of contact plates to recover microbial contaminants from stainless steel surface. MATERIALS AND METHODS: Three commercially available contact plates were used to recover two biological indicators from stainless steel sheets. The method used was standardised and validated to provide robust results. Parameters such as wetness, fertility and loss of water were also investigated for possible correlation with recovery efficiency. RESULTS: The percentage of recovery from the contact plates was low and differences in recovery efficiency between brands depended upon the test organism. The poor recovery was probably due to the inability of the dried micro-organism to transfer to the plate, rather than the inability of the plate to grow the micro-organism. Wetness might help in improving recovery. CONCLUSIONS: The use of a validated protocol allowed robust investigations into the recovery efficiency of contact plates. SIGNIFICANCE AND IMPACT OF THE STUDY: The poor and variable recovery rates are of concern as they cast doubt on the comparability and reliability of environmental monitoring results where different commercial contact plates have been used. PMID- 19187503 TI - Determining the source of Bacillus cereus and Bacillus licheniformis isolated from raw milk, pasteurized milk and yoghurt. AB - AIMS: Strain-specific detection of Bacillus cereus and Bacillus licheniformis in raw and pasteurized milk, and yoghurt during processing. METHODS AND RESULTS: Randomly selected isolates of Bacillus spp. were subjected to PCR analysis, where single primer targeting to the repetitive sequence Box elements was used to fingerprint the species. The isolates were separated into six different fingerprint patterns. The results show that isolates clustered together at about the 57% similarity level with two main groups at the 82% and 83% similarity levels, respectively. Contamination with identical strains both of B. cereus and B. licheniformis in raw and pasteurized milk was found as well as contaminated with different strains (in the case of raw milk and yoghurt/pasteurized milk and yoghurt). Several BOX types traced in processed milk samples were not discovered in the original raw milk. CONCLUSIONS: BOX-PCR fingerprinting is useful for characterizing Bacillus populations in a dairy environment. It can be used to confirm environmental contamination, eventually clonal transfer of Bacillus strains during the technological processing of milk. SIGNIFICANCE AND IMPACT OF THE STUDY: Despite the limited number of strains analysed, the two Bacillus species yielded adequately detectable banding profiles, permitting differentiation of bacteria at the strain level and showing their diversity throughout dairy processing. PMID- 19187504 TI - Cryptosporidium in commercially produced turkeys on-farm and postslaughter. AB - AIMS: To determine the presence of Cryptosporidium species in commercially produced turkey flocks on farm and postslaughter. METHODS AND RESULTS: Three separate turkey flocks were sampled at a single farm and again postslaughter at a commercial processing facility. DNA was extracted and purified from faecal (farm) or caecal (postslaughter) samples and a fragment of 18S rDNA was amplified using a nested PCR approach. Amplified fragments were sequenced, aligned and a neighbour joining tree was constructed. Cryptosporidium meleagridis was not identified in any of the flocks tested. However, all flocks tested positive for Cryptosporidium parvum species. One of the flocks tested positive at the farm and postslaughter. CONCLUSIONS: While C. parvum was present in birds at the farm and postslaughter, turkeys at this facility are not likely to be a significant reservoir for this species. SIGNIFICANCE AND IMPACT OF THE STUDY: Cryptosporidium meleagridis infects avian and human hosts and is increasingly being recognized as a significant human pathogen. However, this study found no evidence of C. meleagridis in commercially produced turkeys at a single location. PMID- 19187505 TI - Evaluation of a digoxigenin-labelled probe for detection of Aeromonas spp. AB - An outer membrane protein-based Digoxigenin (DIG)-labelled DNA probe was developed for the specific detection of Aeromonas sp. from food/environmental/clinical samples. Dot blot reaction answered for all the Aeromonas isolates and was negative for Escherichia coli, Pseudomonas sp., Klebsiella sp., Vibrio parahaemolyticus, V. harveyi, V. alginolyticus, V. vulnificus. Edwardsiella tarda and Staphylococcus sp. As this protein is highly conserved in various Aeromonas species, the probe has the potential for use as a rapid and reliable diagnostic tool. PMID- 19187506 TI - Antimicrobial biosurfactants from marine Bacillus circulans: extracellular synthesis and purification. AB - AIMS: To purify the biosurfactant produced by a marine Bacillus circulans strain and evaluate the improvement in surface and antimicrobial activities. METHODS AND RESULTS: The study of biosurfactant production by B. circulans was carried out in glucose mineral salts (GMS) medium using high performance thin layer chromatography (HPTLC) for quantitative estimation. The biosurfactant production by this strain was found to be growth-associated showing maximum biosurfactant accumulation at 26 h of fermentation. The crude biosurfactants were purified using gel filtration chromatography with Sephadex G-50 matrix. The purification attained by employing this technique was evident from UV-visible spectroscopy and TLC analysis of crude and purified biosurfactants. The purified biosurfactants showed an increase in surface activity and a decrease in critical micelle concentration values. The antimicrobial action of the biosurfactants was also enhanced after purification. CONCLUSIONS: The marine B. circulans used in this study produced biosurfactants in a growth-associated manner. High degree of purification could be obtained by using gel filtration chromatography. The purified biosurfactants showed enhanced surface and antimicrobial activities. SIGNIFICANCE AND IMPACT OF THE STUDY: The antimicrobial biosurfactant produced by B. circulans could be effectively purified using gel filtration and can serve as new potential drugs in antimicrobial chemotherapy. PMID- 19187507 TI - Improved treatment of vulvovaginal candidiasis with fluconazole plus probiotic Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14. AB - AIMS: To determine the ability of probiotic lactobacilli to improve the treatment of vulvovaginal candidiasis (VVC) using a randomized, double-blind and placebo controlled trial. METHODS AND RESULTS: Fifty-five women diagnosed with VVC by vaginal discharge positive for Candida spp. (according to culture method) associated with at least one of the symptoms (itching and burning vaginal feeling, dyspareunia and dysuria), were treated with single dose of fluconazole (150 mg) supplemented every morning for the following 4 weeks with two placebo or two probiotic capsules (containing Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14). At 4 weeks, the probiotic treated group showed significantly less vaginal discharge associated with any of the above mentioned symptoms (10.3%vs 34.6%; P = 0.03) and lower presence of yeast detected by culture (10.3%vs 38.5%; P = 0.014). CONCLUSION: This study has shown that probiotic lactobacilli can increase the effectiveness of an anti-fungal pharmaceutical agent in curing disease. SIGNIFICANCE AND IMPACT OF THE STUDY: This novel finding of probiotic lactobacilli augmenting the cure rate of yeast vaginitis, not only offers an alternative approach to a highly prevalent condition that adversely affects the quality of life of women around the world, but also raises the question of how this combination works. PMID- 19187508 TI - Lactobacillus rhamnosus GG inhibits invasion of cultured human respiratory cells by prtF1-positive macrolide-resistant group A streptococci. AB - AIMS: This study was designed to determine whether the probiotic strain Lactobacillus GG, which is extensively used in the treatment and prevention of intestinal disorders, is able to inhibit invasion of cultured human respiratory cells by macrolide-resistant group A streptococci (GAS) carrying the prtF1 gene, which encodes the fibronectin (Fn)-binding invasin F1. METHODS AND RESULTS: Eight prtF1-positive erythromycin-resistant GAS strains were used to infect A549 monolayers in competition and displacement assays with Lactobacillus GG. Live (L LGG) and heat-killed (HK-LGG) lactobacilli and their spent culture supernatant (SCS) significantly reduced (P < 0.001) GAS invasion efficiency in both assays. No antibacterial activity of Lactobacillus GG against GAS was detected. Both L LGG and HK-LGG and all prtF1-positive GAS induced a strong agglutination reaction using Fn-coated particles. CONCLUSIONS: Lactobacillus GG exerts an antagonistic action against GAS by inhibiting cell invasion. Competitive binding of Lactobacillus GG and GAS to Fn might be involved in the inhibition process. SIGNIFICANCE AND IMPACT OF THE STUDY: The finding that Lactobacillus GG can prevent in vitro invasion of respiratory cells by GAS suggests new applications for this probiotic strain and warrants further studies of its capacity to prevent GAS throat infections. PMID- 19187509 TI - Influence of oligosaccharides on the growth and tolerance capacity of lactobacilli to simulated stress environment. AB - AIMS: Lactobacilli should resist stress environments in industry process and gastrointestinal tract before exerting their beneficial effects. To explore the possible stabilizers in probiotic products, prebiotic oligosaccharides were investigated. METHODS AND RESULTS: We investigated the effect of four selected oligosaccharides on the survival of probiotic Lactobacillus plantarum and L. acidophilus to simulated stress conditions. It was found that the tolerance of lactobacilli to simulated artificial gastrointestinal juice, heat treatment and phenol solution was obviously enhanced in fructo-oligosaccharides (FOS) and xylo oligosaccharides (XOS) group. In addition, chito-oligosaccharides (COS), manno oligosaccharides (MOS) and glucose also had positive effect compared with control group (without sugar). CONCLUSIONS: Prebiotic oligosaccharides, especially XOS and FOS added in medium have protection function to lactobacilli in stress environments. The protection function of oligosaccharides may correlate with the bacteria growth, which was stimulated by these oligosaccharides. SIGNIFICANCE AND IMPACT OF THE STUDY: Prebiotic oligosaccharides may be used as stabilizers in probiotic products. PMID- 19187510 TI - Re-interpretation of the logistic equation for batch microbial growth in relation to Monod kinetics. AB - AIMS: To determine the underlying substrate utilization mechanism in the logistic equation for batch microbial growth by revealing the relationship between the logistic and Monod kinetics. Also, to determine the logistic rate constant in terms of Monod kinetic constants. METHODS AND RESULTS: The logistic equation used to describe batch microbial growth was related to the Monod kinetics and found to be first-order in terms of the substrate and biomass concentrations. The logistic equation constant was also related to the Monod kinetic constants. Similarly, the substrate utilization kinetic equations were derived by using the logistic growth equation and related to the Monod kinetics. CONCLUSION: It is revaled that the logistic growth equation is a special form of the Monod growth kinetics when substrate limitation is first-order with respect to the substrate concentration. The logistic rate constant (k) is directly proportional to the maximum specific growth rate constant (mu(m)) and initial substrate concentration (S(0)) and also inversely related to the saturation constant (K(s)). SIGNIFICANCE AND IMPACT OF THE STUDY: The semi-empirical logistic equation can be used instead of Monod kinetics at low substrate concentrations to describe batch microbial growth using the relationship between the logistic rate constant and the Monod kinetic constants. PMID- 19187511 TI - Conversion of biomass hydrolysates and other substrates to ethanol and other chemicals by Lactobacillus buchneri*. AB - AIMS: A Lactobacillus buchneri strain NRRL B-30929 can convert xylose and glucose into ethanol and chemicals. The aims of the study were to survey three strains (NRRL B-30929, NRRL 1837 and DSM 5987) for fermenting 17 single substrates and to exam NRRL B-30929 for fermenting mixed substrates from biomass hydrolysates. METHODS AND RESULTS: Mixed acid fermentation was observed for all three L. buchneri strains using various carbohydrates; the only exception was uridine which yielded lactate, acetate and uracil. Only B-30929 is capable of utilizing cellobiose, a desired trait in a potential biocatalyst for biomass conversion. Flask fermentation indicated that the B-30929 strain can use all the sugars released from pretreated hydrolysates, and producing 1.98-2.35 g l(-1) ethanol from corn stover hydrolysates and 2.92-3.01 g l(-1) ethanol from wheat straw hydrolysates when supplemented with either 0.25x MRS plus 1% corn steep liquor or 0.5x MRS. CONCLUSIONS: The L. buchneri NRRL B-30929 can utilize mixed sugars in corn stover and wheat straw hydrolysates for ethanol and other chemical production. SIGNIFICANCE AND IMPACT OF THE STUDY: These results are valuable for future research in engineering L. buchneri NRRL B-30929 for fermentative production of ethanol and chemicals from biomass. PMID- 19187512 TI - A polyphasic approach assigns the pathogenic Erwinia strains from diseased pear trees in Japan to Erwinia pyrifoliae. AB - AIMS: Bacterial shoot blight of pear in Japan (BSBP) is caused by Erwinia strains which were formerly associated with the species Erwinia amylovora, the causative agent of fire blight. The description of Erwinia pyrifoliae as a pear pathogen in Korea renewed a possible connection of the pear pathogens in both countries. METHODS AND RESULTS: Nucleotide sequence analysis of the 16S rRNA, the house keeping genes gpd and recA, as well as DNA-DNA hybridization kinetics and microbiological assays place the pear pathogens from Japan into the species E. pyrifoliae described as the causative agent of Asian pear blight in Korea. CONCLUSIONS: Erwinia pyrifoliae strains from Korea and the pear pathogenic Erwinia strains from Japan belong taxonomically into the same species, but show slight divergences in nucleotide sequences used for classification. The allocation is not only supported by microbiological properties, but also by a host range restricted to pear observed before by others. SIGNIFICANCE AND IMPACT OF THE STUDY: The data suggest that the BSBP disease observed at the island of Hokkaido was not fire blight and unify BSBP in Japan with the pear pathogenic species E. pyrifoliae from Korea. PMID- 19187513 TI - Serovar distribution and antimicrobial susceptibility of swine Salmonella isolates from clinically ill pigs in diagnostic submissions from Indiana in the United States. AB - AIMS: To determine serovar distribution and levels of antimicrobial susceptibility of Salmonella isolated from clinically ill pigs in diagnostic submissions. METHODS AND RESULTS: A total of 197 Salmonella isolates were obtained by the Indiana Animal Disease Diagnostic Laboratory from 2003 to 2005. Minimal inhibitory concentrations (MICs) were determined using the standard microbroth dilution method. The top four serovars identified were Salm. enterica serovar Typhimurium variant Copenhagen, Salm. Derby, Salm. Choleraesuis var. Kunzendorf and Salm. Typhimurium. All isolates were susceptible to the fluoroquinolones tested except that eight isolates were intermediate to difloxacin. The isolates showed a low prevalence of resistance to trimethoprim/sulphadiazine (Sxt), gentamicin (G), ceftiofur (Cf) and cephalothin (Cp) with low MIC(50) value of 24 h) (SRQ 1-5: RR 1.4; 95% CI 1.0-1.9, SRQ >or= 6: RR 1.6; 95% CI 1.0-2.6) and delaying initiation of breast-feeding more than eight hours (SRQ 1-5: RR 1.4; 95% CI 0.8 to 2.4, SRQ >or= 6: RR 2.8; 95% CI 1.3-6.1). Worry about delivery was also associated with labour longer than 24 h (RR 1.5; 95% CI 1.0-2.1). CONCLUSIONS: This study provides preliminary evidence of important public health consequences of poor maternal mental health in low-income countries but does not replicate the strong association with low birth weight found in South Asia. PMID- 19187515 TI - Effects of a Skilled Care Initiative on pregnancy-related mortality in rural Burkina Faso. PMID- 19187516 TI - Cluster-sample surveys and lot quality assurance sampling to evaluate yellow fever immunisation coverage following a national campaign, Bolivia, 2007. AB - OBJECTIVE: To estimate the yellow fever (YF) vaccine coverage for the endemic and non-endemic areas of Bolivia and to determine whether selected districts had acceptable levels of coverage (>70%). METHODS: We conducted two surveys of 600 individuals (25 x 12 clusters) to estimate coverage in the endemic and non endemic areas. We assessed 11 districts using lot quality assurance sampling (LQAS). The lot (district) sample was 35 individuals with six as decision value (alpha error 6% if true coverage 70%; beta error 6% if true coverage 90%). To increase feasibility, we divided the lots into five clusters of seven individuals; to investigate the effect of clustering, we calculated alpha and beta by conducting simulations where each cluster's true coverage was sampled from a normal distribution with a mean of 70% or 90% and standard deviations of 5% or 10%. RESULTS: Estimated coverage was 84.3% (95% CI: 78.9-89.7) in endemic areas, 86.8% (82.5-91.0) in non-endemic and 86.0% (82.8-89.1) nationally. LQAS showed that four lots had unacceptable coverage levels. In six lots, results were inconsistent with the estimated administrative coverage. The simulations suggested that the effect of clustering the lots is unlikely to have significantly increased the risk of making incorrect accept/reject decisions. CONCLUSIONS: Estimated YF coverage was high. Discrepancies between administrative coverage and LQAS results may be due to incorrect population data. Even allowing for clustering in LQAS, the statistical errors would remain low. Catch-up campaigns are recommended in districts with unacceptable coverage. PMID- 19187517 TI - Performance of OptiMAL-IT compared to microscopy, for malaria detection in Burkina Faso. AB - OBJECTIVE: To compare the performance of OptiMAL-IT, a rapid diagnostic test for malaria, with that of microscopy in Burkina Faso. METHOD: Finger-prick blood samples of 464 children attending hospital for suspected malaria were tested for malaria by microscopy and OptiMAL-IT. RESULTS: The sensitivity and specificity of OptiMAL-IT were 98.7% (CI 95% = 97.6-99.8) and 96.2% (CI 95% = 94.3-98.1) respectively, with a high positive likelihood ratio (25.97). CONCLUSION: OptiMAL IT can be considered a good method to diagnose malaria in Burkina Faso, particularly in remote areas with little or no access to microscopy services. PMID- 19187518 TI - Cost-effectiveness of artesunate for the treatment of severe malaria. AB - OBJECTIVE: To explore the cost-effectiveness of artesunate against quinine based principally on the findings of a large multi-centre trial carried out in Southeast Asia. METHODS: Trial data were used to compare mortality of patients with severe malaria, treated with either artesunate or quinine. This was combined with retrospectively collected cost data to estimate the incremental cost per death averted with the use of artesunate instead of quinine. RESULTS: The incremental cost per death averted using artesunate was approximately 140 USD. Artesunate maintained this high level of cost-effectiveness also when allowing for the uncertainty surrounding the cost and effectiveness assessments. CONCLUSION: This analysis confirms the vast superiority of artesunate for treatment of severe malaria from an economic as well as a clinical perspective. PMID- 19187519 TI - A new approach to characterize populations of Schistosoma mansoni from humans: development and assessment of microsatellite analysis of pooled miracidia. AB - OBJECTIVES: To develop and assess a microsatellite technique to characterize populations of Schistosoma mansoni from humans. METHODS: For each of five patients, we calculated the allele count and frequency at 11 loci for several pools of miracidia (50 and 100), and compared these to population values, determined by amplifying microsatellites from 186 to 200 individual miracidia per patient. RESULTS: We were able to detect up to 94.5% of alleles in pools. Allele count and frequency strongly and significantly correlated between singles and pools; marginally significant differences (P < 0.05) were detected for one patient (pools of 50) for allele frequencies and for two patients (pools of 100) for allele counts. Kato-Katz egg counts and number of alleles per pool did not co vary, indicating that further direct comparisons of the results from these two techniques are needed. CONCLUSIONS: Allele counts and frequency profiles from pooling provide important information about infection intensity and complexity, beyond that obtained from traditional methods. Although we are not advocating use of pooling to replace individual genotyping studies, it can potentially be useful in certain applications as a rapid and cost effective screening method for studies of S. mansoni population genetics, or as a more informative way to quantify and characterize human worm populations. PMID- 19187520 TI - Infants with severe neonatal jaundice in Lagos, Nigeria: incidence, correlates and hearing screening outcomes. AB - OBJECTIVES: To establish the incidence, correlates and hearing screening outcomes of infants with severe neonatal jaundice in Nigeria. METHODS: Community-based study in which all infants attending Bacille Calmette-Guerin immunisation clinics in inner-city Lagos were enrolled into a universal hearing screening programme during which correlates of severe neonatal jaundice (requiring phototherapy and/or exchange blood transfusion) were explored with multivariable logistic regression. RESULTS: Of the 5262 infants enrolled, only 48.7% were born in hospitals although almost all mothers (97.9%) attended antenatal clinics. 6.7% had a history of neonatal jaundice of whom 5.5% (95% CI:4.9-6.2) received phototherapy and 1.9% (95% CI:1.5-2.3) had an exchange blood transfusion. Factors independently associated with severe neonatal jaundice were maternal religion, occupation, use of herbal preparations during pregnancy, infant's gender, weight at screening, multiple gestation and place of birth. All but two infants with severe neonatal jaundice were exclusively breast-fed. Of those who failed the hearing tests, 17.3% were treated with phototherapy and 11.3% had an exchange blood transfusion. At least 8.9% of infants requiring phototherapy and 17.3% of those requiring exchange blood transfusion were at risk of sensorineural hearing loss. CONCLUSIONS: Severe neonatal jaundice is a significant condition associated with modifiable risk factors in this population. Policy initiatives for prevention, early detection followed by appropriate and timely intervention are urgently needed to reduce the disease burden. PMID- 19187521 TI - In-vivo efficacy of amodiaquine-artesunate in children with uncomplicated Plasmodium falciparum malaria in western Kenya. AB - OBJECTIVES: To assess the efficacy of amodiaquine-artesunate in an area with high chloroquine resistance in western Kenya. METHODS: Twenty-eight day in-vivo efficacy trial of amodiaquine-artesunate in 103 children aged 6-59 months in western Kenya with smear-confirmed uncomplicated Plasmodium falciparum malaria. RESULTS: The 28-day uncorrected adequate clinical and parasitological response (ACPR) was 69.0%, with 15.5% Late Clinical Failure and 15.5% Late Parasitologic Failure rates. The PCR-corrected 28-day ACPR was 90.2%. Clinical risk factors for recurrent infection (recrudescences and reinfections) were lower axillary temperature at enrollment and low weight-for-age Z-score. The presence of single nucleotide polymorphisms pfcrt 76T and pfmdr1 86Y at baseline was associated with increased risk of recurrent infections, both reinfections and recrudescences. CONCLUSION: Although artemether-lumefantrine (Coartem) is the first line ACT in Kenya, amodiaquine-artesunate is registered as an option for treatment of uncomplicated P. falciparum and remains an effective alternative to Coartem in western Kenya. Continued amodiaquine monotherapy in the private sector may jeopardize the future use of amodiaquine-artesunate as an alternative artemisinin based combination therapy. PMID- 19187522 TI - Early assessment of the implementation of a national programme for the prevention of mother-to-child transmission of HIV in Cameroon and the effects of staff training: a survey in 70 rural health care facilities. AB - OBJECTIVES: To assess the availability of equipment and the staff's knowledge to prevent Mother-To-Child Transmission (PMTCT) in rural healthcare facilities recently covered by the national PMTCT programme in Cameroon. METHODS: In eight districts inventories of antiviral drugs and HIV test kits were made on site, using a standardised check-list. Knowledge of HIV and PMTCT was evaluated with a multiple-choice (MC) questionnaire based on typical clinical PMTCT cases. Staff participated subsequently in a 2-day training on HIV/AIDS and the Cameroon PMTCT guidelines. Immediately after training and after 7 months, retention of knowledge was tested with the same questions but in different order and layout. RESULTS: Sixty two peripheral nurse-led clinics and the eight district hospitals were assessed. Whereas all district hospitals presented complete equipment, only six of the peripheral clinics (10%) were equipped with both complete testing materials and a full set of drugs to provide PMTCT. Thirty six peripheral facilities (58%) possessed full equipment for HIV-testing and 8 (13%) stocked all PMTCT drugs. Of 137 nurses, 102 (74%) agreed to the two knowledge tests. Fewer than 66% knew that HIV-diagnosis requires positive results in two different types of rapid tests and only 19% chose the right recommendation on infant-feeding for HIV-positive mothers. Correct answers on drug regimens in different PMTCT settings varied from 25% to 56%. All percentages of correct answers improved greatly with training (P < 0.001) and retention remained high 7 months after training (P < 0.001). CONCLUSIONS: Prevent Mother-To-Child Transmission programmes in settings such as rural Cameroon need to be adapted to the special needs of peripheral nurse-led clinics. Appropriate short training may considerably improve nurses' competence in PMTCT. Other important components are regular supervision and measures to guarantee supply of equipment in rural areas. PMID- 19187523 TI - Design of epidemiological studies measuring genital and plasma HIV-1 outcomes: lessons from a randomised controlled trial. AB - OBJECTIVE: HIV-1 genital viral loads have not been extensively used as markers of HIV transmissibility. We set out to determine whether the variability of genital HIV-1 RNA over time necessitates design adjustments in studies measuring genital shedding to account for this variability. METHODS: We used data from a completed trial of HSV suppressive therapy to estimate the correlation of plasma and genital HIV-1 RNA quantities sampled at different times. These correlation estimates were used to estimate the relative sample sizes needed to detect an impact on HIV-1 genital and plasma quantities assuming a variable number of pre- and post-randomisation repeated measurements. The treatment effect on quantities of genital and plasma HIV-1 RNA were analysed using random effects linear regression. RESULTS: Post-randomisation plasma HIV-1 RNA was highly correlated within-women, while genital HIV-1 RNA was less strongly correlated. Related to this, the sample size required to detect a treatment effect on genital HIV-1 RNA decreased with increasing numbers of post-randomisation measurements up to 6-7 measurements, but varied less for plasma HIV-1 RNA. In contrast, repeated pre randomisation measurements of plasma HIV-1 RNA increased study power more than genital HIV-1 RNA because of the high correlation of plasma HIV-1 RNA measurements between the pre- and post-randomisation samples. Re-analysis of the trial data illustrated the increased precision of the treatment effect on genital HIV-1 with increasing post-randomisation measurements. CONCLUSIONS: Designs allowing for repeated post-randomisation measures should be used to increase the precision in estimates of genital HIV-1 RNA. Repeated post-randomisation measurements of plasma HIV-1 RNA are of limited benefit. PMID- 19187524 TI - Socio-environmental predictors of Barmah forest virus transmission in coastal areas, Queensland, Australia. AB - OBJECTIVE: To assess the socio-environmental predictors of Barmah forest virus (BFV) transmission in coastal areas, Queensland, Australia. METHODS: Data on BFV notified cases, climate, tidal levels and socioeconomic index for area (SEIFA) in six coastal cities, Queensland, for the period 1992-2001 were obtained from the relevant government agencies. Negative binomial regression models were used to assess the socio-environmental predictors of BFV transmission. RESULTS: The results show that maximum and minimum temperature, rainfall, relative humidity, high and low tide were statistically significantly associated with BFV incidence at lags 0-2 months. The fitted negative binomial regression models indicate a significant independent association of each of maximum temperature (beta = 0.139, P = 0.000), high tide (beta = 0.005, P = 0.000) and SEIFA index (beta = -0.010, P = 0.000) with BFV transmission after adjustment for confounding variables. CONCLUSIONS: The transmission of BFV disease in Queensland coastal areas seemed to be determined by a combination of local social and environmental factors. The model developed in this study may have applications in the control and prevention of BFV disease in these areas. PMID- 19187525 TI - Evolutionary lability of odour-mediated host preference by the malaria vector Anopheles gambiae. AB - Many species of disease-vector mosquitoes display vertebrate host specificity. Despite considerable progress in recent years in understanding the proximate and ultimate factors related to non-random host selection at the interspecific level, the basis of this selection remains only partially understood. Anopheles gambiae sensu stricto, the main malaria vector in Africa, is considered a highly anthropophilic mosquito, and host odours have been shown to play a major role in the host-seeking process of this species. Studies on host preference of An. gambiae have been either conducted in controlled conditions using laboratory reared mosquitoes and worn stockings as host-related stimuli, or have been done in the field with methods that do not account for internal (e.g. age of sampled mosquitoes) and/or environmental effects. We explored differential behavioural responses to host odours between two populations of the same sibling species, An. gambiae in semi-field conditions in Burkina Faso. The behavioural responses (i.e. degree of activation and strength of anemotaxis) were investigated using a Y olfactometer designed to accommodate whole hosts as a source of odour stimuli. Two strains of An. gambiae (3 to 4-day-old female) from laboratory Kisumu strain, and from field-collected individuals were confronted to combinations of stimuli comprising calf odour, human odour and outdoor air. In dual-choice tests, field mosquitoes chose human odour over calf odour, outdoor air over calf odour and responded equally to human and outdoor air, while laboratory mosquitoes responded equally to human and calf odour, human odour over outdoor air and calf odour over outdoor air. Overall, no effect of CO(2) exhaled by humans and calves neither on the proportion of activated mosquitoes nor on the relative attractiveness to odour stimuli was found. We report for the first time an intraspecific variation in host-odour responses. This study clearly suggests that there may be genetic polymorphism underlying host preference and emphasizes that the highly anthropophilic label given to An. gambiae s.s. must be carefully interpreted and refer to populations rather than the whole sibling species. PMID- 19187526 TI - MCRS2 represses the transactivation activities of Nrf1. AB - BACKGROUND: Nrf1 [p45 nuclear factor-erythroid 2 (p45 NF-E2)-related factor 1], a member of the CNC-bZIP (CNC basic region leucine zipper) family, is known to be a transcriptional activator by dimerization with distinct partners, such as Maf, FosB, c-Jun, JunD, etc. The transcriptional roles of CNC-bZIP family are demonstrated to be involved in globin gene expression as well as the antioxidant response. For example, CNC-bZIP factors can regulate the expression of detoxification proteins through AREs, such as expression of human gamma glutamylcysteine synthetases (GCS), glutathione S-transferases (GST), UDP glucuronosyl transferase (UDP-GT), NADP (H) quinone oxidoreductase (NQOs), etc. To further explore other factor(s) in cells related to the function of Nrf1, we performed a yeast two-hybrid screening assay to identify any Nrf1-interacting proteins. In this study, we isolated a cDNA encoding residues 126-475 of MCRS2 from the HeLa cell cDNA library. Some functions of MCRS1 and its splice variant MSP58 and MCRS2 have been previously identified, such as transforming, nucleolar sequestration, ribosomal gene regulation, telomerase inhibition activities, etc. Here, we demonstrated MCRS2 can function as a repressor on the Nrf1-mediated transactivation using both in vitro and in vivo systems. RESULTS: To find other proteins interacting with the CNC bZIP domain of Nrf1, the CNC-bZIP region of Nrf1 was used as a bait in a yeast two-hybrid screening assay. MCRS2, a splicing variant of p78/MCRS1, was isolated as the Nrf1-interacting partner from the screenings. The interaction between Nrf1 and MCRS2 was confirmed in vitro by GST pull-down assays and in vivo by co-immunoprecipitation. Further, the Nrf1-MCRS2 interaction domains were mapped to the residues 354-447 of Nrf1 as well as the residues 314-475 of MCRS2 respectively, by yeast two-hybrid and GST pull-down assays. By immunofluorescence, MCRS2-FLAG was shown to colocalize with HA-Nrf1 in the nucleus and didn't result in the redistribution of Nrf1. This suggested the existence of Nrf1-MCRS2 complex in vivo. To further confirm the biological function, a reporter driven by CNC-bZIP protein binding sites was also shown to be repressed by MCRS2 in a transient transfection assay. An artificial reporter gene activated by LexA-Nrf1 was also specifically repressed by MCRS2. CONCLUSION: From the results, we showed MCRS2, a new Nrf1-interacting protein, has a repression effect on Nrf1-mediated transcriptional activation. This was the first ever identified repressor protein related to Nrf1 transactivation. PMID- 19187527 TI - Hilbert-Huang versus Morlet wavelet transformation on mismatch negativity of children in uninterrupted sound paradigm. AB - BACKGROUND: Compared to the waveform or spectrum analysis of event-related potentials (ERPs), time-frequency representation (TFR) has the advantage of revealing the ERPs time and frequency domain information simultaneously. As the human brain could be modeled as a complicated nonlinear system, it is interesting from the view of psychological knowledge to study the performance of the nonlinear and linear time-frequency representation methods for ERP research. In this study Hilbert-Huang transformation (HHT) and Morlet wavelet transformation (MWT) were performed on mismatch negativity (MMN) of children. Participants were 102 children aged 8-16 years. MMN was elicited in a passive oddball paradigm with duration deviants. The stimuli consisted of an uninterrupted sound including two alternating 100 ms tones (600 and 800 Hz) with infrequent 50 ms or 30 ms 600 Hz deviant tones. In theory larger deviant should elicit larger MMN. This theoretical expectation is used as a criterion to test two TFR methods in this study. For statistical analysis MMN support to absence ratio (SAR) could be utilized to qualify TFR of MMN. RESULTS: Compared to MWT, the TFR of MMN with HHT was much sharper, sparser, and clearer. Statistically, SAR showed significant difference between the MMNs elicited by two deviants with HHT but not with MWT, and the larger deviant elicited MMN with larger SAR. CONCLUSION: Support to absence ratio of Hilbert-Huang Transformation on mismatch negativity meets the theoretical expectations, i.e., the more deviant stimulus elicits larger MMN. However, Morlet wavelet transformation does not reveal that. Thus, HHT seems more appropriate in analyzing event-related potentials in the time-frequency domain. HHT appears to evaluate ERPs more accurately and provide theoretically valid information of the brain responses. PMID- 19187528 TI - Biological activity of a genetically modified BMP-2 variant with inhibitory activity. AB - BACKGROUND: Alterations of the binding epitopes of bone morphogenetic protein-2 (BMP-2) lead to a modified interaction with the ectodomains of BMP receptors. In the present study the biological effect of a BMP-2 double mutant with antagonistic activity was evaluated in vivo. METHODS: Equine-derived collagenous carriers were loaded with recombinant human BMP-2 (rhBMP-2) in a well-known dose to provide an osteoinductive stimulus. The study was performed in a split animal design: carriers only coupled with rhBMP-2 (control) were implanted into prepared cavities of lower limb muscle of rats, specimens coupled with rhBMP-2 as well as BMP-2 double mutant were placed into the opposite limb in the same way. After 28 days the carriers were explanted, measured radiographically and characterized histologically. RESULTS: As expected, the BMP-2 loaded implants showed a typical heterotopic bone formation. The specimens coupled with both proteins showed a significant decreased bone formation in a dose dependent manner. CONCLUSION: The antagonistic effect of a specific BMP-2 double mutant could be demonstrated in vivo. The dose dependent influence on heterotopic bone formation by preventing rhBMP-2 induced osteoinduction suggests a competitive receptor antagonism. PMID- 19187529 TI - Ancient, independent evolution and distinct molecular features of the novel human T-lymphotropic virus type 4. AB - BACKGROUND: Human T-lymphotropic virus type 4 (HTLV-4) is a new deltaretrovirus recently identified in a primate hunter in Cameroon. Limited sequence analysis previously showed that HTLV-4 may be distinct from HTLV-1, HTLV-2, and HTLV-3, and their simian counterparts, STLV-1, STLV-2, and STLV-3, respectively. Analysis of full-length genomes can provide basic information on the evolutionary history and replication and pathogenic potential of new viruses. RESULTS: We report here the first complete HTLV-4 sequence obtained by PCR-based genome walking using uncultured peripheral blood lymphocyte DNA from an HTLV-4-infected person. The HTLV-4(1863LE) genome is 8791-bp long and is equidistant from HTLV-1, HTLV-2, and HTLV-3 sharing only 62-71% nucleotide identity. HTLV-4 has a prototypic genomic structure with all enzymatic, regulatory, and structural proteins preserved. Like STLV-2, STLV-3, and HTLV-3, HTLV-4 is missing a third 21-bp transcription element found in the long terminal repeats of HTLV-1 and HTLV-2 but instead contains unique c-Myb and pre B-cell leukemic transcription factor binding sites. Like HTLV-2, the PDZ motif important for cellular signal transduction and transformation in HTLV-1 and HTLV-3 is missing in the C-terminus of the HTLV-4 Tax protein. A basic leucine zipper (b-ZIP) region located in the antisense strand of HTLV-1 and believed to play a role in viral replication and oncogenesis, was also found in the complementary strand of HTLV-4. Detailed phylogenetic analysis shows that HTLV-4 is clearly a monophyletic viral group. Dating using a relaxed molecular clock inferred that the most recent common ancestor of HTLV-4 and HTLV-2/STLV-2 occurred 49,800 to 378,000 years ago making this the oldest known PTLV lineage. Interestingly, this period coincides with the emergence of Homo sapiens sapiens during the Middle Pleistocene suggesting that early humans may have been susceptible hosts for the ancestral HTLV-4. CONCLUSION: The inferred ancient origin of HTLV-4 coinciding with the appearance of Homo sapiens, the propensity of STLVs to cross-species into humans, the fact that HTLV-1 and -2 spread globally following migrations of ancient populations, all suggest that HTLV-4 may be prevalent. Expanded surveillance and clinical studies are needed to better define the epidemiology and public health importance of HTLV-4 infection. PMID- 19187530 TI - Evolution and diversity of Rickettsia bacteria. AB - BACKGROUND: Rickettsia are intracellular symbionts of eukaryotes that are best known for infecting and causing serious diseases in humans and other mammals. All known vertebrate-associated Rickettsia are vectored by arthropods as part of their life-cycle, and many other Rickettsia are found exclusively in arthropods with no known secondary host. However, little is known about the biology of these latter strains. Here, we have identified 20 new strains of Rickettsia from arthropods, and constructed a multi-gene phylogeny of the entire genus which includes these new strains. RESULTS: We show that Rickettsia are primarily arthropod-associated bacteria, and identify several novel groups within the genus. Rickettsia do not co-speciate with their hosts but host shifts most often occur between related arthropods. Rickettsia have evolved adaptations including transmission through vertebrates and killing males in some arthropod hosts. We uncovered one case of horizontal gene transfer among Rickettsia, where a strain is a chimera from two distantly related groups, but multi-gene analysis indicates that different parts of the genome tend to share the same phylogeny. CONCLUSION: Approximately 150 million years ago, Rickettsia split into two main clades, one of which primarily infects arthropods, and the other infects a diverse range of protists, other eukaryotes and arthropods. There was then a rapid radiation about 50 million years ago, which coincided with the evolution of life history adaptations in a few branches of the phylogeny. Even though Rickettsia are thought to be primarily transmitted vertically, host associations are short lived with frequent switching to new host lineages. Recombination throughout the genus is generally uncommon, although there is evidence of horizontal gene transfer. A better understanding of the evolution of Rickettsia will help in the future to elucidate the mechanisms of pathogenicity, transmission and virulence. PMID- 19187531 TI - Potential determinants of obesity among children and adolescents in Germany: results from the cross-sectional KiGGS Study. AB - BACKGROUND: Obesity among children and adolescents is a growing public health problem. The aim of the present paper is to identify potential determinants of obesity and risk groups among 3- to 17-year old children and adolescents to provide a basis for effective prevention strategies. METHODS: Data were collected in the German Health Interview and Examination Survey for Children and Adolescents (KiGGS), a nationally representative and comprehensive data set on health behaviour and health status of German children and adolescents. Body height and weight were measured and body mass index (BMI) was classified according to IOTF cut-off points. Statistical analyses were conducted on 13,450 non-underweight children and adolescents aged 3 to 17 years. The association between overweight, obesity and several potential determinants was analysed for this group as well as for three socio-economic status (SES) groups. A multiple logistic regression model with obesity as the dependent variable was also calculated. RESULTS: The strongest association with obesity was observed for parental overweight and for low SES. Furthermore, a positive association with both overweight (including obesity) and obesity was seen for maternal smoking during pregnancy, high weight gain during pregnancy (only for mothers of normal weight), high birth weight, and high media consumption. In addition, high intakes of meat and sausages, total beverages, water and tea, total food and beverages, as well as energy-providing food and beverages were significantly associated with overweight as well as with obesity. Long sleep time was negatively associated with obesity among 3- to 10-year olds. Determinants of obesity occurred more often among children and adolescents with low SES. CONCLUSION: Parental overweight and a low SES are major potential determinants of obesity. Families with these characteristics should be focused on in obesity prevention. PMID- 19187532 TI - A noncoding RNA gene on chromosome 10p15.3 may function upstream of hTERT. AB - BACKGROUND: We attempted to clone candidate genes on 10p 14-15 which may regulate hTERT expression, through exon trapping using 3 BAC clones covering the region. After obtaining 20 exons, we examined the function of RGM249 (RGM: RNA gene for miRNAs) we cloned from primary cultured human hepatocytes and hepatoma cell lines. We confirmed approximately 20 bp products digested by Dicer, and investigated the function of this cloned gene and its involvement in hTERT expression by transfecting the hepatoma cell lines with full-length dsRNA, gene specific designed siRNA, and shRNA-generating plasmid. RESULTS: RGM249 showed cancer-dominant intense expression similar to hTERT in cancer cell lines, whereas very weak expression was evident in human primary hepatocytes without telomerase activity. This gene was predicted to be a noncoding precursor RNA gene. Interestingly, RGM249 dsRNA, siRNA, and shRNA inhibited more than 80% of hTERT mRNA expression. In contrast, primary cultured cells overexpressing the gene showed no significant change in hTERT mRNA expression; the overexpression of the gene strongly suppressed hTERT mRNA in poorly differentiated cells. CONCLUSION: These findings indicate that RGM249 might be a microRNA precursor gene involved in the differentiation and function upstream of hTERT. PMID- 19187533 TI - Structural studies of the Enterococcus faecalis SufU [Fe-S] cluster protein. AB - BACKGROUND: Iron-sulfur clusters are ubiquitous and evolutionarily ancient inorganic prosthetic groups, the biosynthesis of which depends on complex protein machineries. Three distinct assembly systems involved in the maturation of cellular Fe-S proteins have been determined, designated the NIF, ISC and SUF systems. Although well described in several organisms, these machineries are poorly understood in Gram-positive bacteria. Within the Firmicutes phylum, the Enterococcus spp. genus have recently assumed importance in clinical microbiology being considered as emerging pathogens for humans, wherein Enterococcus faecalis represents the major species associated with nosocomial infections. The aim of this study was to carry out a phylogenetic analysis in Enterococcus faecalis V583 and a structural and conformational characterisation of it SufU protein. RESULTS: BLAST searches of the Enterococcus genome revealed a series of genes with sequence similarity to the Escherichia coli SUF machinery of [Fe-S] cluster biosynthesis, namely sufB, sufC, sufD and SufS. In addition, the E. coli IscU ortholog SufU was found to be the scaffold protein of Enterococcus spp., containing all features considered essential for its biological activity, including conserved amino acid residues involved in substrate and/or co-factor binding (Cys50,76,138 and Asp52) and, phylogenetic analyses showed a close relationship with orthologues from other Gram-positive bacteria. Molecular dynamics for structural determinations and molecular modeling using E. faecalis SufU primary sequence protein over the PDB:1su0 crystallographic model from Streptococcus pyogenes were carried out with a subsequent 50 ns molecular dynamic trajectory. This presented a stable model, showing secondary structure modifications near the active site and conserved cysteine residues. Molecular modeling using Haemophilus influenzae IscU primary sequence over the PDB:1su0 crystal followed by a MD trajectory was performed to analyse differences in the C terminus region of Gram-positive SufU and Gram-negative orthologous proteins, in which several modifications in secondary structure were observed. CONCLUSION: The data describe the identification of the SUF machinery for [Fe-S] cluster biosynthesis present in the Firmicutes genome, showing conserved sufB, sufC, sufD and sufS genes and the presence of the sufU gene coding for scaffold protein, instead of sufA; neither sufE nor sufR are present. Primary sequences and structural analysis of the SufU protein demonstrated its structural-like pattern to the scaffold protein IscU nearby on the ISC machinery. E. faecalis SufU molecular modeling showed high flexibility over the active site regions, and demonstrated the existence of a specific region in Firmicutes denoting the Gram positive region (GPR), suggested as a possible candidate for interaction with other factors and/or regulators. PMID- 19187534 TI - Characterization of Cronobacter recovered from dried milk and related products. AB - BACKGROUND: Cronobacter is a recently proposed genus consisting of six genomospecies that encompass the organisms previously identified as Enterobacter sakazakii. Cronobacter are opportunistic pathogens and are known to cause serious infections in infants, particularly neonates. High case fatality rates have been associated with infections and acute sequelae can occur in survivors with severe ramifications on neurological development. Infant formula has been identified as one route of transmission for infection in infants. However, the primary reservoirs for subsequent contamination of foods with Cronobacter remain undefined due to the ubiquitous nature of these organisms. More recently, infections in adults have been reported, especially amongst the elderly and patients who are immunocompromised. To help prevent the transmission of infection, it is important to identify the main food sources for Cronobacter. The aim of this study was to identify and characterize Cronobacter isolated from dried-milk and related products available in an Egyptian food market. RESULTS: In total sixteen Cronobacter strains were isolated from 152 dairy-based products. These were identified and characterized using pheno- and genotyping experiments. Real-time PCR confirmed the detection of Cronobacter. Following antibiotic susceptibility tests, 3 strains showed resistance to trimethoprim and/or neomycin. Phenotype profiles were generated based on key biochemical distinguishing tests. Pulsed-field gel electrophoresis (PFGE) identified 8 PFGE types amongst the collection of strains. Repetitive sequence based PCR (rep-PCR) analysis identified 3 rep-PCR types amongst the collection of strains. Sequencing of the recN gene was used to differentiate among the recently described species of Cronobacter. CONCLUSION: This study identified the presence of Cronobacter in dried milk and related products sourced from the Nile-Delta region of Egypt. Although the majority of the strains were susceptible to the antibiotics tested, resistance was observed in three isolates, highlighting the risks associated with Cronobacter contamination in foods. Phenotype and genotype analysis should be applied to further characterize Cronobacter spp. and prevent its transmission into food products. PMID- 19187535 TI - Diagnosis and management of retroperitoneal ancient schwannomas. AB - BACKGROUND: Ancient schwannomas are degenerate peripheral nerve sheath tumors that very rarely occur in the retroperitoneum. They generally reach large proportions before producing symptoms due to mass effect. We describe three cases of retroperitoneal ancient schwannomas and discuss the diagnosis and management of these tumors. CASE PRESENTATIONS: Three female patients with retroperitoneal ancient schwannomas were reviewed. One patient presented with several weeks of upper abdominal pain and lower chest discomfort, whereas back pain and leg pain with associated weakness were predominant symptoms in the remaining two. Abdominal imaging findings demonstrated heterogeneous masses in the retroperitoneum with demarcated margins, concerning for malignancy. The patients successfully had radical excision of their tumors. Histological examination showed encapsulated tumors that displayed alternating areas of dense cellularity and areas of myxoid matrix consistent with a diagnosis of ancient schwannoma. CONCLUSION: A diagnosis of ancient schwannoma should be entertained for any heterogeneous, well encapsulated mass in the retroperitoneum. In these cases less radical surgical resection should be considered as malignant transformation of these tumors is extremely rare and recurrence is uncommon following excision. PMID- 19187536 TI - Safety and benefits of large-volume liposuction: a single center experience. AB - BACKGROUND: Liposuction is a surgical technique to remove excess fat deposits from specific areas of the body. Purpose of this study is to determine how far large volume liposuction is safe and effective. METHODOLOGY: From July 2003 to December 2005, 60 female patients had liposuction of different areas of the body as waist, hips, buttocks, thighs, and knees. Their mean age was 30.6 +/- 15.4 years old. A standard liposuction technique was done by using a tumescent infiltration formula. The average amount of infusate was 3000 cc, with an average aspirate amount of 6000 cc. Pre-operative anthropometric measurements as weight, height, body mass index, areas to be liposuctioned in addition to pre-operative hematological investigations as complete blood picture, blood sugar, liver function tests, blood urea, serum creatinine, and serum cholesterol were done. RESULTS: The results were evaluated with preoperative and postoperative photographs. Postoperative anthropometric measurements and hematological investigations were done at 6th week, and 4th month after surgery. The rate of complications was low and relatively minor in nature. No major complications were presented. Minor complications have occurred as skin irregularities (20%), Seroma (15%), Garment pressure sore (10%), Cutaneous hyper-pigmentation (5%). CONCLUSION: Large-volume liposuction can be performed safely and it can produce desirable morphological and hematological changes. PMID- 19187538 TI - The effects of stochastic resonance electrical stimulation and neoprene sleeve on knee proprioception. AB - BACKGROUND: A variety of knee injuries and pathologies may cause a deficit in knee proprioception which may increase the risk of reinjury or the progression of disease. Stochastic resonance stimulation is a new therapy which has potential benefits for improving proprioceptive function. The objective of this study was to determine if stochastic resonance (SR) stimulation applied with a neoprene sleeve could improve knee proprioception relative to a no-stimulation/no-sleeve condition (control) or a sleeve alone condition in the normal, healthy knee. We hypothesized that SR stimulation when applied with a sleeve would enhance proprioception relative to the control and sleeve alone conditions. METHODS: Using a cross-over within subject design, twenty-four healthy subjects were tested under four combinations of conditions: electrical stimulation/sleeve, no stimulation/sleeve, no stimulation/no sleeve, and stimulation/no sleeve. Joint position sense (proprioception) was measured as the absolute mean difference between a target knee joint angle and the knee angle reproduced by the subject. Testing was conducted during both partial-weight bearing (PWB) and non-weight bearing (NWB) tasks. Differences in joint position sense between the conditions were evaluated by repeated-measures analysis of variance testing. RESULTS: Joint position sense error during the stimulation/sleeve condition (2.48 degrees +/- 1.32 degrees ) was found to be more accurate (P < 0.05) relative to the control condition (3.35 degrees +/- 1.63 degrees ) in the PWB task. No difference in joint position sense error was found between stimulation/sleeve and sleeve alone conditions for the PWB task. Joint position sense error was not found to differ between any of the conditions for the NWB task. CONCLUSION: These results suggest that SR electrical stimulation when combined with a neoprene sleeve is an effective modality for enhancement of joint proprioception in the PWB knee. We believe these results suggest the need for further study of the potential of SR stimulation to correct proprioceptive deficits in a clinical population with knee injury/pathology or in subjects at risk of injury because of a proprioceptive deficit. PMID- 19187537 TI - Gene expression profiling of the tumor microenvironment during breast cancer progression. AB - INTRODUCTION: The importance of the tumor microenvironment in breast cancer has been increasingly recognized. Critical molecular changes in the tumor stroma accompanying cancer progression, however, remain largely unknown. We conducted a comparative analysis of global gene expression changes in the stromal and epithelial compartments during breast cancer progression from normal to preinvasive to invasive ductal carcinoma. METHODS: We combined laser capture microdissection and gene expression microarrays to analyze 14 patient-matched normal epithelium, normal stroma, tumor epithelium and tumor-associated stroma specimens. Differential gene expression and gene ontology analyses were performed. RESULTS: Tumor-associated stroma undergoes extensive gene expression changes during cancer progression, to a similar extent as that seen in the malignant epithelium. Highly upregulated genes in the tumor-associated stroma include constituents of the extracellular matrix and matrix metalloproteases, and cell-cycle-related genes. Decreased expression of cytoplasmic ribosomal proteins and increased expression of mitochondrial ribosomal proteins were observed in both the tumor epithelium and the stroma. The transition from preinvasive to invasive growth was accompanied by increased expression of several matrix metalloproteases (MMP2, MMP11 and MMP14). Furthermore, as observed in malignant epithelium, a gene expression signature of histological tumor grade also exists in the stroma, with high-grade tumors associated with increased expression of genes involved in immune response. CONCLUSIONS: Our results suggest that the tumor microenvironment participates in tumorigenesis even before tumor cells invade into stroma, and that it may play important roles in the transition from preinvasive to invasive growth. The immune cells in the tumor stroma may be exploited by the malignant epithelial cells in high-grade tumors for aggressive invasive growth. PMID- 19187539 TI - Mapping of IgE-binding regions on recombinant Cyn d 1, a major allergen from Bermuda Grass Pollen (BGP). AB - BACKGROUND: Bermuda grass (Cynodon dactylon; subfamily Chloridoideae) is an important source of seasonal aeroallergens in warm tropical and sub-tropical areas worldwide. Improved approaches to diagnosis and therapy of allergic diseases require a thorough understanding of the structure and epitopes on the allergen molecule that are crucial for the antigen-antibody interaction. This study describes the localization of the human IgE-binding regions of the major group 1 pollen allergen Cyn d 1 from Bermuda grass. METHODS: A cDNA library was constructed from Bermuda grass pollen (BGP) using a Lambda gt11 expression vector. The gene encoding the Cyn d 1 allergen was isolated by screening the library with a mouse monoclonal antibody raised against grass group 1 allergen. In order to characterize the IgE epitopes on Cyn d 1, seven overlapping fragments and three deletion mutants were cloned and over-expressed in E. coli. The recombinant fragments and deletion mutants were evaluated for their comparative IgE reactivity with sera of non atopic individuals and grass pollen allergic patients by ELISA and a dot-blot assay. RESULTS: Analysis of IgE binding regions by overlapping fragments and deletion mutants identified two major allergenic regions corresponding to amino acids 120-170 and 224-244. Deletion of either or both regions led to a significant reduction in IgE binding, emphasizing the importance of the C-terminal region on Cyn d 1 in epitope-IgE interaction. CONCLUSION: Anti-Cyn d 1 IgE antibodies from allergic human sera recognize two epitopes located at the C-terminal end of the molecule. These data will enable the design of improved diagnostic and therapeutic approaches for BGP hypersensitivity. PMID- 19187540 TI - Bilateral variant testicular arteries with double renal arteries. AB - BACKGROUND: The testicular arteries normally arise from the abdominal aorta. There are reports about the variant origin of these arteries. Accessory renal arteries are also a common finding but their providing origin to testicular arteries is an important observation. The variations described here are unique and provide significant information to surgeons dissecting the abdominal cavity. CASE PRESENTATION: During routine dissection classes of abdominal region of a 60 year-old male cadaver, we observed bilateral variant testicular arteries and double renal arteries. CONCLUSION: Awareness of variations of the testicular arteries such as those presented here becomes important during surgical procedures like varicocele and undescended testes. PMID- 19187541 TI - Issues associated with assessing nuclear localization of N-terminally unphosphorylated beta-catenin with monoclonal antibody 8E7. AB - BACKGROUND: Beta-catenin is a dual function adhesion/transcriptional co-activator protein, and both functions are critical for normal tissue homeostasis. Since the transcriptional functions of beta-catenin are more often implicated in various disease processes, there is much interest in the development and use of reagents to interrogate spatial and temporal evidence of beta-catenin nuclear signaling in cells and tissues. An important study demonstrated that the signaling form of beta-catenin is specifically unphosphorylated at residues S37 and T41, and suggested that this form exhibits a propensity for cytosolic/nuclear accumulation relative to the total pool of beta-catenin. RESULTS: We show that monoclonal antibody, 8E7, which recognizes the signaling form of beta-catenin specifically unphosphorylated at S37 and T41 (Active B-Catenin, ABC), also cross-reacts with a widely expressed, variably accessible nuclear antigen that is not beta-catenin. In cell types commonly used to study Wnt activation, this non-specific nuclear staining can be robust, obscuring the ABC signal. Definitive detection of nuclear localized ABC can be confirmed through an ability of classical cadherins to sequester ABC to cell junctions. In tissues, milder antigen retrieval methods can reduce the accessibility of mAb 8E7 to this cross-reacting nuclear antigen. CONCLUSION: These findings reveal that interpretation of nuclear, signaling active beta-catenin using monoclonal antibody 8E7 should be considered judiciously, and in conjunction with independent methods. PMID- 19187542 TI - FIP1L1-PDGFRA molecular analysis in the differential diagnosis of eosinophilia. AB - BACKGROUND: Primary eosinophlia associated with the FIP1L1-PDGFRA rearrangement represents a subset of chronic eosinophilic leukaemia (CEL) and affected patients are very sensitive to imatinib treatment. This study was undertaken in order to examine the prevalence and the associated clinicopathologic and genetic features of FIP1L1-PDGFRA rearrangement in a cohort of 15 adult patients presenting with profound eosinophilia (> 1.5 x 109/L). METHODS: Reverse transcriptase-polymerase chain reaction (RT-PCR) was used for the detection of FIP1L1-PDGFRA rearrangement and the results confirmed by direct sequencing. C-KIT-D816V mutation was analysed retrospectively by PCR and restriction-fragment-length-polymorphism (PCR-RFLP), in all cases with primary eosinophilia. RESULTS: Two male patients with splenomegaly carried the FIP1L1-PDGFRA rearrangement, whilst 2 others were ultimately classified as suffering from idiopathic hypereosinophlic syndrome (HES) and one from systemic mastocytosis. These patients were negative for the C KIT-D816V mutation and received imatinib (100-400 mg daily). Patients with CEL and HES responded to imatinib and remained in complete haematological, clinical and molecular (for carriers of FIP1L1-PDGFRA rearrangement) remission for a median of 28.2 months (range: 11-54), whilst the patient with systemic mastocytosis did not respond. Interestingly, in both patients with FIP1L1-PDGFRA rearrangement, the breakpoints into PDGFRA were located within exon 12 and fused with exons 8 and 8a of FIP1L1, respectively. CONCLUSION: An early diagnosis of FIPIL1-PDGFRA-positive CEL and imatinib treatment offer to the affected patients an excellent clinical therapeutic result, avoiding undesirable morbidity. Moreover, although the molecular mechanisms underlying disease pathogenesis remain to be determined, imatinib can be effective in patients with idiopathic HES. PMID- 19187543 TI - Radiation produces differential changes in cytokine profiles in radiation lung fibrosis sensitive and resistant mice. AB - BACKGROUND: Recent research has supported that a variety of cytokines play important roles during radiation-induced lung toxicity. The present study is designed to investigate the differences in early cytokine induction after radiation in sensitive (C57BL/6) and resistant mice (C3H). RESULTS: Twenty-two cytokines in the lung tissue homogenates, bronchial lavage (BAL) fluids, and serum from 3, 6, 12, 24 hrs to 1 week after 12 Gy whole lung irradiation were profiled using a microsphere-based multiplexed cytokine assay. The majority of cytokines had similar baseline levels in C57BL/6 and C3H mice, but differed significantly after radiation. Many, including granulocyte colony-stimulating factor (G-CSF), interleukin-6 (IL-6), and keratinocyte-derived chemokine (KC) were elevated significantly in specimens from both strains. They usually peaked at about 3-6 hrs in C57BL/6 and 6-12 hrs in C3H. At 6 hrs in lung tissue, G-CSF, IL-6, and KC increased 6, 8, and 11 fold in C57BL/6 mice, 4, 3, and 3 fold in the C3H mice, respectively. IL-6 was 10-fold higher at 6 hrs in the C57BL/6 BAL fluid than the C3H BAL fluid. MCP-1, IP-10, and IL-1alpha also showed some differences between strains in the lung tissue and/or serum. For the same cytokine and within the same strain of mice, there were significant linear correlations between lung tissue and BAL fluid levels (R2 ranged 0.46-0.99) and between serum and tissue (R2 ranged 0.56-0.98). CONCLUSION: Radiation induced earlier and greater temporal changes in multiple cytokines in the pulmonary fibrosis sensitive mice. Positive correlation between serum and tissue levels suggests that blood may be used as a surrogate marker for tissue. PMID- 19187544 TI - Detection of genetically modified organisms (GMOs) using isothermal amplification of target DNA sequences. AB - BACKGROUND: The most common method of GMO detection is based upon the amplification of GMO-specific DNA amplicons using the polymerase chain reaction (PCR). Here we have applied the loop-mediated isothermal amplification (LAMP) method to amplify GMO-related DNA sequences, 'internal' commonly-used motifs for controlling transgene expression and event-specific (plant-transgene) junctions. RESULTS: We have tested the specificity and sensitivity of the technique for use in GMO studies. Results show that detection of 0.01% GMO in equivalent background DNA was possible and dilutions of template suggest that detection from single copies of the template may be possible using LAMP. CONCLUSION: This work shows that GMO detection can be carried out using LAMP for routine screening as well as for specific events detection. Moreover, the sensitivity and ability to amplify targets, even with a high background of DNA, here demonstrated, highlights the advantages of this isothermal amplification when applied for GMO detection. PMID- 19187545 TI - Selecting control genes for RT-QPCR using public microarray data. AB - BACKGROUND: Gene expression analysis has emerged as a major biological research area, with real-time quantitative reverse transcription PCR (RT-QPCR) being one of the most accurate and widely used techniques for expression profiling of selected genes. In order to obtain results that are comparable across assays, a stable normalization strategy is required. In general, the normalization of PCR measurements between different samples uses one to several control genes (e.g. housekeeping genes), from which a baseline reference level is constructed. Thus, the choice of the control genes is of utmost importance, yet there is not a generally accepted standard technique for screening a large number of candidates and identifying the best ones. RESULTS: We propose a novel approach for scoring and ranking candidate genes for their suitability as control genes. Our approach relies on publicly available microarray data and allows the combination of multiple data sets originating from different platforms and/or representing different pathologies. The use of microarray data allows the screening of tens of thousands of genes, producing very comprehensive lists of candidates. We also provide two lists of candidate control genes: one which is breast cancer-specific and one with more general applicability. Two genes from the breast cancer list which had not been previously used as control genes are identified and validated by RT-QPCR. Open source R functions are available at http://www.isrec.isb sib.ch/~vpopovic/research/ CONCLUSION: We proposed a new method for identifying candidate control genes for RT-QPCR which was able to rank thousands of genes according to some predefined suitability criteria and we applied it to the case of breast cancer. We also empirically showed that translating the results from microarray to PCR platform was achievable. PMID- 19187546 TI - Shadows of the colonial past--diverging plant use in Northern Peru and Southern Ecuador. AB - This paper examines the traditional use of medicinal plants in Northern Peru and Southern Ecuador, with special focus on the Departments of Piura, Lambayeque, La Libertad, Cajamarca, and San Martin, and in Loja province, with special focus on the development since the early colonial period. Northern Peru represents the locus of the old Central Andean "Health Axis." The roots of traditional healing practices in this region go as far back as the Cupisnique culture early in the first millennium BC. Northern Peru and Southern Ecuador share the same cultural context and flora but show striking differences in plant use and traditional knowledge. Two hundred fifteen plant species used for medicinal purposes in Ecuador and 510 plant species used for medicinal purposes in Peru were collected, identified,. and their vernacular names, traditional uses, and applications recorded. This number of species indicates that the healers, market vendors, and members of the public interviewed in Peru still have a very high knowledge of plants in their surroundings, which can be seen as a reflection of the knowledge of the population in general. In Ecuador much of the original plant knowledge has already been lost. In Peru, 433 (85%) were Dicotyledons, 46 (9%) Monocotyledons, 21 (4%) Pteridophytes, and 5 (1%) Gymnosperms. Three species of Giartina (Algae) and one species of the Lichen genus Siphula were used. The families best represented were Asteraceae with 69 species, Fabaceae (35), Lamiaceae (25), and Solanaceae (21). Euphorbiaceae had 12 species, and Poaceae and Apiaceae each accounted for 11 species. In Ecuador the families best represented were Asteraceae (32 species), Euphorbiaceae, Lamiaceae, and Solanaceae (11 species each), and Apiaceae, Fabaceae, Lycopodiaceae (9 species each). One hundred eighty two (85%) of the species used were Dicotyledons, 20 Monocotyledons (9.3%), 12 ferns (5.5%), and one unidentified lichen was used. Most of the plants used (83%) were native to Peru and Ecuador. Fresh plants, often collected wild, were used in two thirds of all cases in Peru, but in almost 95% of the cases in Ecuador. The most common applications included the ingestion of herb decoctions or the application of plant material as poultices. Although about 50% of the plants in use in the colonial period have disappeared from the popular pharmacopoeia, the overall number of plant species used medicinally has increased in Northern Peru, while Southern Ecuador shows a decline of plant knowledge since colonial times. PMID- 19187547 TI - Relationship between heart rate recovery and inflammatory markers in patients with polycystic ovary syndrome: a cross-sectional study. AB - BACKGROUND: Polycystic ovary syndrome (PCOS) is an endocrine disease closely related to several risk factors for cardiovascular disease. An abnormal heart rate recovery (HRR), an easily-obtained measure derived from exercise stress test and closely related to an increased risk for cardiovascular mortality, has been recently described in PCOS women. A subclinical increase of the inflammation markers has been also observed in the PCOS. This study was designed to study the relationships between HRR and inflammatory markers in PCOS women. METHODS: Two hundred forty-three young PCOS patients without known risk factors for cardiovascular risk were enrolled. All patients underwent hormonal and metabolic profile, white blood cells (WBCs) count and C-reactive protein (CRP). HRR was calculated as the difference between heart rate at peak exercise and heart rate at first minute of the cool-down period. Abnormal HRR was defined as 10000 fraction of BRL CM accelerated the development of the embryos (90.3%). This beneficial effect was also evident even in the absence of EDTA. RT-PCR analysis revealed that mRNAs encoding the beta-A or beta-B subunit of activins (Mr ~29000), which are well characterized cytokines that act as releasers of the 2-cell block, were expressed in BRL cells. These results indicate that BRL cells synthesize Fr.B-25 at low levels, and that activins contained in the BRL CM probably contributed to overcoming the 2-cell block of CD-1 zygotes cultured in vitro. PMID- 19187568 TI - Evaluating integrated healthcare for refugees and hosts in an African context. AB - This paper argues on ethical and practical grounds for more widespread use of an integrated approach to refugee healthcare, and proposes a basic model of assessment for integrated systems. A defining element of an integrated approach is an equal ability by refugee and host nationals to access the same healthcare resources from the same providers. This differs fundamentally from parallel care, currently the predominant practice in Africa. The authors put forward a general model for evaluation of integrated healthcare with four criteria: (1) improved health outcomes for both hosts and refugees, (2) increased social integration, (3) increased equitable use of healthcare resources, and (4) no undermining of protection. Historical examples of integrated care in Ethiopia and Uganda are examined in light of these criteria to illustrate how this evaluative model would generate evidence currently lacking in debates on the merit of integrated healthcare. PMID- 19187569 TI - Access as a policy-relevant concept in low- and middle-income countries. AB - Although access to health care is frequently identified as a goal for health care policy, the precise meaning of access to health care often remains unclear. We present a conceptual framework that defines access to health care as the empowerment of an individual to use health care and as a multidimensional concept based on the interaction (or degree of fit) between health care systems and individuals, households, and communities. Three dimensions of access are identified: availability, affordability, and acceptability, through which access can be evaluated directly instead of focusing on utilisation of care as a proxy for access. We present the case for the comprehensive evaluation of health care systems as well as the dimensions of access, and the factors underlying each dimension. Such systemic analyses can inform policy-makers about the 'fit' between needs for health care and receipt of care, and provide the basis for developing policies that promote improvements in the empowerment to use care. PMID- 19187570 TI - Access to health care services--an English policy perspective. AB - The English government has given a commitment to improving access to health care services for particular groups perceived as being under-served, or served inappropriately, by existing services. In this article four examples of policies aimed at improving access are considered: enhancing the supply of services to under-served areas, changing the organization of services, setting targets to improve access, and empowering people to make choices. Policies aimed at improving access will work only if they address the source of inequities, which means identifying the key barriers to access and these barriers are unlikely to be uniform across sectors, services, and groups of people. Evidence on the success of these four types of intervention in terms of influencing access and equity of access is discussed, borrowing some concepts from the sociological literature that enable us to understand the importance of how barriers to access may arise for different services and different population groups. It is clear that some policies may not work as well as we would hope, or may even exacerbate inequities of access, because they fail to recognize the source of the particular barriers faced by some groups. PMID- 19187571 TI - The bases of identification: when optimal distinctiveness needs face social identity threat. AB - Three studies were conducted to examine the predictions that (a) in-group identification depends on optimal distinctiveness needs (Study 1), and (b) that social identity threat overrides the predictive value of these needs to determine identification (Studies 2 and 3). In Study 1, need for assimilation and need for differentiation were assessed among natural groups. We found support for the optimal distinctiveness theory (ODT) prediction that there is a curvilinear relationship between identification and optimal distinctiveness needs satisfaction. In Studies 2 and 3, interactive effects of the extent to which groups satisfy assimilation and differentiation needs (groups are either too small, of moderate size, or too large) and social identity threat were examined. In the no identity threat condition identification was higher in moderately sized groups (where both needs are balances) compared to groups were either assimilation or differentiation dominates. However, when facing an identity threat, identification was highest in very small groups, providing evidence that social identity concerns override individual need satisfaction. Discussion focuses on comparing and integrating ODT and social identity theory. PMID- 19187572 TI - The effects of subjective norms on behaviour in the theory of planned behaviour: a meta-analysis. AB - A meta-analysis investigated the effects of perceived injunctive (IN) and descriptive (DN) norms on behaviour (BEH) within the theory of planned behaviour (TPB) in a sample of 196 studies. Two related correlation matrices (pairwise and listwise) were synthesized from the data and used to model the TPB relations with path analyses. Convergent evidence indicated that the relation between DN and BEH was stronger than the relation between IN and BEH. Evidence also suggested a significant direct relation between DN and BEH in the context of TPB. A suppressor effect of IN on DN in its relation with BEH was also noted. Moderator analyses indicated that the DN-BEH relation was stronger when there was more time between measures of cognition and behaviour, when behaviours were not socially approved, more socially motive and more pleasant; results were mixed in the case of the IN-BEH relation. Results imply that IN and DN are conceptually different constructs. PMID- 19187573 TI - Illegitimacy and identity threat in (inter)action: predicting intergroup orientations among minority group members. AB - We test the hypothesis that intergroup orientations among minority group members are shaped by the interaction between the perceived illegitimacy of intergroup relations and identity threat appraisals, as well as their main effects. This is because together they serve to focus emotion-mediated reactions on the out group's role in threatening in-group identity. In a large-scale field study (N=646), conducted among the Welsh minority in the UK, we quasi-manipulated the extent to which Welsh identity was dependent on the 'threatened' Welsh language. Results supported our hypothesis that the illegitimacy x identity threat interaction would be strongest where Welsh identity was most dependent upon the Welsh language, and through intergroup anger would predict support for more radical, unconstitutional forms of action. PMID- 19187574 TI - A Box-Cox normal model for response times. AB - The log-transform has been a convenient choice in response time modelling on test items. However, motivated by a dataset of the Medical College Admission Test where the lognormal model violated the normality assumption, the possibilities of the broader class of Box-Cox transformations for response time modelling are investigated. After an introduction and an outline of a broader framework for analysing responses and response times simultaneously, the performance of a Box Cox normal model for describing response times is investigated using simulation studies and a real data example. A transformation-invariant implementation of the deviance information criterium (DIC) is developed that allows for comparing model fit between models with different transformation parameters. Showing an enhanced description of the shape of the response time distributions, its application in an educational measurement context is discussed at length. PMID- 19187575 TI - An interpretative phenomenological analysis of living with chronic low back pain. AB - OBJECTIVES: A predominantly quantitative approach to the psychological study of chronic low back pain (CLBP) has shown that enduring negative emotional responses and passive coping strategies contribute towards disability. The main objective of this study was to extend existing knowledge by providing a detailed and contextualized understanding of the meaning of CLBP for participants with long standing experiences of chronic pain. DESIGN: This is a qualitative, semi structured interview study. The data were analysed using interpretative phenomenological analysis (IPA). This is the first of three sets of individual interviews comprising a longitudinal study of the same participants over 3 years. METHODS: Ten patients with CLBP were interviewed prior to their attendance at a medically staffed chronic pain clinic. The sample comprised seven females and three males. Ages ranged between 39 and 66 years. All had experienced CLBP for at least 4 years. CONCLUSIONS: The participants' experiences are represented by three main themes: 'maintaining integrity', 'the crucial nature of the pain' and 'managing the pain' that highlight participants' understanding of their pain within a biomedical framework. The findings are discussed with reference to the role of illness beliefs in the management of CLBP. PMID- 19187576 TI - Obesity and health-related quality of life: does social support moderate existing associations? AB - OBJECTIVES: Obesity has been shown to be negatively related to physical health related quality of life (HQOL) much more strongly than mental HQOL. This is remarkable given findings on obesity-related social stigmata and associations with depression. Considering obesity as a stressor, this study tests for a moderating role of social support for obesity/HQOL associations among women and men. DESIGN: Data come from N=2,732 participants aged 35-74 years in a 2004-2005 general population survey in the Augsburg region, Germany. METHODS: Body weight and height were assessed by anthropometric measurements (classified by body mass index using WHO standards), social support by the Social Support Questionnaire 14 item Short-Form (F-SozU-K14) and HQOL by the 12-item Short-Form Health Survey (SF 12). In multiple regression and general linear models, age, education, family status, health insurance, and place of residence were adjusted for. RESULTS: Among both genders, obesity was associated with reduced physical but not mental HQOL. Among men reporting strong social support, physical HQOL was impaired neither in the moderately nor the severely obese group (compared with normal weight), while it was given less social support. Among women, poor physical HQOL was associated with obesity regardless of social support. CONCLUSIONS: In this adult population sample, no association was found for obesity with mental HQOL. In contrast, a negative association with physical HQOL exists for all subgroups except men with strong social support, indicating that social support buffers obesity-related impairments in physical HQOL in men but not in women. This suggests that obese women and men with strong social support represent distinct populations, with possible implications for obesity care. PMID- 19187577 TI - Coping with examinations: exploring relationships between students' coping strategies, implicit theories of ability, and perceived control. AB - BACKGROUND: Relatively little is known about the contribution of students' beliefs regarding the nature of academic ability (i.e. their implicit theories) on strategies used to deal with examinations. AIMS: This study applied Dweck's socio-cognitive model of achievement motivation to better understand how students cope with examinations. It was expected that students' implicit theories of academic ability would be related to their use of particular coping strategies to deal with exam-related stress. Additionally, it was predicted that perceived control over exams acts as a mediator between implicit theories of ability and coping. SAMPLE: Four hundred and ten undergraduate students (263 males, 147 females), aged from 17 to 26 years old (M=19.73, SD=1.46) were volunteers for the present study. METHODS: Students completed measures of coping, implicit theories of academic ability, and perception of control over academic examinations during regular classes in the first term of the university year. RESULTS: Multiple regression analyses revealed that incremental beliefs of ability significantly and positively predicted active coping, planning, venting of emotions, seeking social support for emotional and instrumental reasons, whereas entity beliefs positively predicted behavioural disengagement and negatively predicted active coping and acceptance. In addition, analyses revealed that entity beliefs of ability were related to coping strategies through students' perception of control over academic examinations. CONCLUSIONS: These results confirm that exam-related coping varies as a function of students' beliefs about the nature of academic ability and their perceptions of control when approaching examinations. PMID- 19187578 TI - Does emotion predict the course of major depressive disorder? A review of prospective studies. AB - PURPOSE: Emotional dysfunction is a hallmark of major depressive disorder (MDD). There has been wide interest in identifying the nature and significance of emotional deficits in MDD. Given the paramount goal of identifying etiological markers of MDD that can sharpen prevention and treatment efforts, researchers have pursued emotion as a functionally significant factor and predictor of clinical course in MDD. This review is the first to summarize the literature examining emotion as a predictor of the clinical course of MDD. METHOD: We conducted a systematic review of all published studies reporting on the prospective relationship between positive and negative emotionality (PE and NE)- measured at the state or trait level--and the longitudinal course of MDD in diagnosed adults. Physiological, behavioural, and subjective indices of emotion were considered. The primary analyses encompassed 28 research reports that included data from 3,798 participants and tested a total of 60 hypotheses. RESULTS: Lower levels of PE predicted poorer MDD course with some consistency. Paradoxically, both lower levels of state NE and higher levels of trait NE predicted poorer MDD course. The relationships between emotionality and MDD course generally held even after initial depression symptom severity was taken into account. CONCLUSION: Emotion shows promise as a predictor of MDD course. Implications of these data for current theories of emotion and MDD, the apparently divergent relation between state and trait NE and course, and future directions to further clarify the functional significance of emotion in the context of MDD are discussed. PMID- 19187579 TI - Visuospatial working memory interference with recollections of trauma. AB - OBJECTIVES: Laboratory research using a working memory framework has shown modality-specific reductions in image vividness and emotionality when concurrent tasks are performed while maintaining the image in consciousness. We extended this research to trauma images in a clinical population awaiting treatment for post-traumatic stress symptoms. DESIGN: A within-subjects design was used, with each participant completing an imagery task under three concurrent task conditions: side-to-side eye-movements, counting, and exposure only (no concurrent task). METHODS: Eighteen participants selected three images each, the images being those that were the most distressing from participants' trauma memories and most likely to intrude involuntarily. Participants gave baseline ratings of the vividness and emotionality of each of their trauma images. Each image was assigned to a condition. Each condition comprised 8 trials in which participants recollected the appropriate image for 8 s while performing eye movements, counting or no concurrent task, and then rated its vividness and emotionality. Follow-up ratings were obtained by telephone 1 week later. RESULTS: The eye-movement task reduced vividness and emotionality of the trauma images relative to the counting task and exposure only, but did so only during the imagery period and not at follow-up. The images were predominantly visual. CONCLUSIONS: Concurrent tasks matched to the modality of trauma images may provide a useful treatment aid for temporarily dampening emotional responses to recollections of trauma. PMID- 19187580 TI - Cross-sectional study of congenital heart disease among Tibetan children aged from 4 to 18 years at different altitudes in Qinghai Province. AB - BACKGROUND: Congenital heart disease (CHD) is a common heart condition which does considerable harm to the health of children and adolescents. CHD epidemiological characteristics of Tibetan children whose ages ranged from 4 to 18 years were investigated in Qinghai Province. METHODS: A total of 32 578 Tibetan children, living at altitudes of 2535 m, 3600 m and 4200 m, were examined using a three stage protocol: prescreening, rechecking and diagnosis using a color Doppler. The distribution of CHD at different altitudes was analyzed together with differences in occurrence according to age and gender. RESULTS: A total of 235 CHD cases were discovered. The total prevalence of CHD was 7.21 per thousand. Prevalence of CHD has been shown to increase along with increase in altitude with 5.45 per thousand at an altitude of 2535 m, 6.80 per thousand at 3600 m and 9.79 per thousand at 4200 m. There were no statistically significant differences between the prevalence at 2535 m and 3600 m (chi(2) = 1.594, P > 0.05). However, there was a significant difference between the prevalence at 2535 m and 4200 m (chi(2) = 7.002, P < 0.01). Also, apparent differences existed between the prevalence at 3600 m and at 4200 m (chi(2) = 5.540, P < 0.05). There was no statistically significant difference in prevalence according to age at an altitude of 2535 m, but the rate of CHD increased significantly along with increasing age at 3600 m and 4200 m. The total prevalence ratio of children aged from 16 to 18 years was significantly higher than that of children from 4 to 7, and from 8 to 12 with chi(2) values of 10.79 (P < 0.005), and 5.60 (P < 0.05) respectively. Within the constituent ratio of CHD, the prevalence of atrial septal defect (ASD) was the highest at 39.10%, followed by the prevalence of ventricular septal defect (VSD) at 32.8% and patent ductus arteriosus (PDA) at 24.7%. Furthermore the proportion of the four categories of CHD varied at different altitude levels: at 2535 m, of those diagnosed with CHD, the prevalence rate of VSD was the highest at 43.5%, at 3600 m ASD was the highest at 42.8% and at 4200 m, PDA was the highest at 50.8%. CONCLUSION: The epidemiological characteristics of CHD in Tibetan children may be associated with altitude levels. PMID- 19187581 TI - Early neurosurgical intervention of spinal cord contusion: an analysis of 30 cases. AB - BACKGROUND: The incidence of spinal injury with spinal cord contusion is high in developed countries and is now growing in China. Furthermore, spinal cord injury happens mostly in young people who have a long life expectance. A large number of patients thus are wheelchair bound for the rest of their lives. Therefore, spinal cord injury has aroused great concern worldwide. Despite great efforts, recovery from spinal cord injury remains unsatisfactory. Based on the pathology of spinal cord contusion, an idea of early neurosurgical intervention has been formulated in this study. METHODS: A total of 30 patients with "complete" spinal cord injury or classified as American Spinal Injury Association (ASIA)-A were studied. Orthopedic treatment of the injured vertebra (e), internal fixation of the vertebral column, and bilateral laminectomy for epidural decompression were followed directly by neurosurgical management, including separation of the arachnoid adhesion to restore cerebrospinal fluid flow and debridement of the spinal cord necrotic tissue with concomitant intramedullary decompression. Rehabilitation started 17 days after the operation. The final outcome was evaluated after 3 months of rehabilitation. Pearson chi-square analysis was used for statistical analysis. RESULTS: All the patients recovered some ability to walk. The least recovered patients were able to walk with a wheeled weight support and help in stabilizing the weight bearing knee joint (12 cases, 40%). Thirteen patients (43%) were able to walk with a pair of crutches, a stick or without any support. The timing of the operation after injury was important. An optimal operation time window was identified at 4 - 14 days after injury. CONCLUSIONS: Early neurosurgical intervention of spinal cord contusion followed by rehabilitation can significantly improve the locomotion of the patients. It is a new idea of a therapeutic approach for spinal cord contusion and has been proven to be very successful. PMID- 19187582 TI - Screening of tumor suppressor genes on 1q31.1-32.1 in Chinese patients with sporadic colorectal cancer. AB - BACKGROUND: As a model for both multistep and multipathway carcinogenesis, colorectal neoplastic progression provides paradigms for researching both oncogenes and tumor suppressor genes (TSGs). However, the mechanism of colorectal cancer (CRC) is not completely understood, and many genes may be involved in the colorectal carcinogenesis. The purpose of this study was to screen for the potential TSGs on chromosome 1q31.1-32.1 in Chinese patients with sporadic colorectal cancer, to explore whether colorectal cancer in the Chinese population has unique genetic alterations and determine whether other putative TSGs exist and contribute to colon carcinogenesis. METHODS: Six polymorphic microsatellite markers, at a density of approximately one marker in every 1.6 cM, were chosen for refined loss of heterozygosity (LOH) mapping of 1q31.1-32.1. Eighty-three colorectal cancer patients' tumor and normal DNA were analyzed via polymerase chain reaction (PCR) for these microsatellite markers. PCR products were eletrophoresed on an ABI 377 DNA sequencer. Genescan 3.1 and Genotype 2.1 software were used for LOH scanning and analysis. On the basis of refined LOH mapping results, we undertook a microarray-based expression screening to identify tumor association genes in 19 of the CRC cases. RESULTS: The average LOH frequency of 1q31.1-32.1 was 24.41%, with the highest frequency of 36.73% (18/49) at D1S2622, and the lowest of 16.42% (11/67) at D1S412. A minimal region of frequent deletion was located within a 2 cM genomic segment at D1S413-D1S2622. There was no significant association between LOH of any marker in the studied regions and the clinicopathological data (patient sex, age, tumor size, growth pattern, or Dukes stage). On the basis of refined mapping results, we chose 25 genes located in the D1S413-D1S2622 (1q31.3-32.1) region and presented a microarray-based high throughput screening approach in 19 sporadic CRC cases to identify candidate CRC related tumor suppressor genes. This study found 4 significantly down-expressed genes, including CSRP1, LMOD1, PPP1R12B and CFHL3. There was no significant association between expression levels of CFHL3, CSRP1, LMOD1, PPP1R12B and the clinicopathological data. By database searching, CSRP1 was hypothesized to be a colorectal cancer related tumor suppressor gene. CONCLUSIONS: Through detailed deletion mapping, we found that the 1q31.3-32.1 region might harbor one or more colorectal cancer related tumor suppressor gene (s). And by microarray-based high-throughput screening of candidate genes located in this region and by subsequent database searching, we present the first evidence that CSRP1 might be involved in the progression of CRC. PMID- 19187583 TI - Multi-slice CT contrast-enhanced presentations of advanced gastric cancer: associations with histo-differentiation and expression of p53 and P-glycoprotein. AB - BACKGROUND: This study aimed to investigate multi-slice CT contrast-enhanced presentation of gastric cancer and its correlation with histo-differentiation and p53 and P-glycoprotein (P-gp) expression. METHODS: Sixty-six patients with gastric cancer in the present study underwent a multi-slice CT preoperative routine and dual-phase contrast-enhanced examination of the upper abdomen; postoperative specimens were used to determine histo-differentiation and the expression of p53 and P-gp. The correlation of multi-slice CT contrast-enhanced presentation with histo-differentiation and expression of p53 and P-gp was analyzed. RESULTS: The dual-phase contrast-enhanced ratio (CER) was not correlated with the histo-differentiation of gastric cancer (P > 0.05). Positive expression of p53 and P-gp was significantly higher in the cases of layered or heterogeneous enhancement than in the cases of homogenous enhancement (P < 0.05). Positive expression of p53 was also correlated with the arterial phase CER, tumor size and lymph node metastasis (P < 0.05), but not with infiltration thickness of the gastric wall, nor was it correlated with the portal phase CER (P > 0.05). Positive expression of P-gp was only correlated with the portal phase CER (P = 0.005). CONCLUSIONS: Differently enhanced pattern and CER of the arterial and portal phase in gastric cancer correlate with its different histo-differentiation and expression of p53 and P-gp respectively. In addition, tumor size and lymph node metastasis of gastric cancer relate to the expression of p53. PMID- 19187584 TI - Identification of differential gene expression for microarray data using recursive random forest. AB - BACKGROUND: The major difficulty in the research of DNA microarray data is the large number of genes compared with the relatively small number of samples as well as the complex data structure. Random forest has received much attention recently; its primary characteristic is that it can form a classification model from the data with high dimensionality. However, optimal results can not be obtained for gene selection since it is still affected by undifferentiated genes. We proposed recursive random forest analysis and applied it to gene selection. METHODS: Recursive random forest, which is an improvement of random forest, obtains optimal differentiated genes after step by step dropping of genes which, according to a certain algorithm, have no effects on classification. The method has the advantage of random forest and provides a gene importance scale as well. The value of the area under the curve (AUC) of the receiver operating characteristic (ROC) curve, which synthesizes the information of sensitivity and specificity, is adopted as the key standard for evaluating the performance of this method. The focus of the paper is to validate the effectiveness of gene selection using recursive random forest through the analysis of five microarray datasets; colon, prostate, leukemia, breast and skin data. RESULTS: Five microarray datasets were analyzed and better classification results have been attained using only a few genes after gene selection. The biological information of the selected genes from breast and skin data was confirmed according to the National Center for Biotechnology Information (NCBI). The results prove that the genes associated with diseases can be effectively retained by recursive random forest. CONCLUSIONS: Recursive random forest can be effectively applied to microarray data analysis and gene selection. The retained genes in the optimal model provide important information for clinical diagnoses and research of the biological mechanism of diseases. PMID- 19187585 TI - Hemodynamic variables and clinical features correlated with serum uric acid in patients with pulmonary arterial hypertension. AB - BACKGROUND: Serum uric acid (UA), the final product of purine degradation, has been proposed to be a marker for the severity and a possible predictor of mortality in patients with pulmonary arterial hypertension (PAH). The objectives of this study were to elucidate whether serum UA level correlates with the clinical features and the hemodynamic variables in Chinese patients with PAH and to compare the difference of the correlates in patients associated with different etiologies. METHODS: Serum UA was assessed in 228 patients with three types of PAH (idiopathic PAH (IPAH), congenital heart disease related PAH (CHD-PAH) and connective tissue disease related PAH (CTD-PAH)) together with other clinical features. After the individualized treatment for at least 6 months, the UA levels and clinical features were re-evaluated in 88 patients. RESULTS: Serum UA was significantly elevated in patients with PAH compared with age-matched control subjects ((350.40 +/- 108.73) micromol/L vs (266.91 +/- 81.38) micromol/L), P < 0.001). Serum UA negatively correlated with cardiac output and mixed venous saturation (SvO(2)) in all three types of PAH (all P < 0.05), positively correlated with the size of right ventricle in IPAH (P = 0.002) and CTD-PAH (P = 0.013) patients and with pulmonary vascular resistance just in CTD-PAH patients (P = 0.001). Serum UA significantly decreased from (365.80 +/- 120.46) micromol/L to (333.67 +/- 117.56) micromol/L in 88 patients (P = 0.006) with vasodilator therapy for at least 6 months, accompanied with a reduction in pulmonary vascular resistance from (15.13 +/- 6.96) Woods unit to (12.00 +/- 5.04) Woods unit (P = 0.001) and an increase in cardiac output from (2.63 +/- 0.98) L/min to (3.08 +/- 1.04) L/min (P = 0.005). CONCLUSIONS: Serum UA increases in proportion to the clinical severity of all the three types of PAH, especially the CTD-PAH had a stronger correlations compared with IPAH and CHD-PAH. The serum UA levels also could partly reflect the response to the treatment in patients with PAH. PMID- 19187586 TI - Acute coronary syndrome is an independent risk factor for late incomplete stent apposition after sirolimus-eluting stent implantation. AB - BACKGROUND: Late incomplete stent apposition (ISA) may occur after drug-eluting stent implantation, affecting long-term clinical outcomes. This study aimed to evaluate the impact of clinical presentations of coronary artery disease on late ISA after percutaneous coronary intervention (PCI) with sirolimus-eluting stents (SES) by means of three-dimensional volumetric intravascular ultrasound (IVUS) analyses. METHODS: One hundred and thirty-seven patients with coronary artery disease received SES implantation during PCI and had repeat angiography with IVUS examination. All patients were followed up one year after the procedure. RESULTS: In overall 219 treated lesions (137 patients), late ISA was identified in 25 lesions (16 patients). Clinical diagnosis of acute coronary syndrome (ACS) and use of long stents were more common in patients with than in those without late ISA. Patients with late ISA had greater external elastic membrane (EEM) area in stented segment ((15.34 +/- 5.44) vs (13.83 +/- 4.51) mm(2), P = 0.026), stented to-reference segment EEM area ratio (1.13 +/- 0.22 vs 1.02 +/- 0.18, P < 0.001), and plaque and media area ((8.43 +/- 3.93) vs (7.01 +/- 2.93) mm(2), P = 0.002) than in those without late ISA. Multivariate Logistic regression analysis showed that clinical diagnosis of ACS and use of long stents were independent risk factors for late ISA (OR 6.477, 95% CI 2.297 - 18.263, P < 0.001; OR 3.680, 95% CI 1.181 - 11.469, P = 0.025; respectively). During one-year follow-up after IVUS examination, the rate of very late stent thrombosis tended to be higher in patients with than in those without late ISA (18.7% vs 3.3%, P = 0.051). CONCLUSIONS: The occurrence of late ISA after SES implantation may be related to clinical status, use of long stents, and marked positive vessel remodeling. Late ISA tended to increase the rate of very late stent thrombosis during follow-up, highlighting the importance of long-term dual antiplatelet therapy for these patients. PMID- 19187588 TI - Objective evaluation of burn and post-surgical scars and the accuracy of subjective scar type judgment. AB - BACKGROUND: The true proliferation degree of burn and operation scars during their maturation was poorly defined. The objective of this study was to provide a quantitative evaluation of the proliferation degree of deep-partial thickness burn and surgical suture wound scars after wound healing and the accuracy of subjective judgment of scar types. METHODS: A total of 423 patients in the burn group and 667 patients in the suture wound group were enrolled in this study. Objective scar measurement (by negative-positive moulage) was carried out at 3, 6 and 12 months after wound healing in 1090 patients from different regions in China. The Kruskal-Wallis H test and chi-square test were used in the statistical analysis. RESULTS: The median degrees of scar proliferation were 0.03 cm at 3 months, 0.03 cm at 6 months and 0 cm at 12 months in the burn group and 0.05 cm, 0.04 cm and 0.04 cm respectively in the suture wound group. At 3 and 6 months post wound healing, there was no significant difference in scar proliferation between the groups (P > 0.01). After 1 year, burn scars displayed evidence of atrophy (P < 0.01). Although the degree of scar proliferation differed among the four scar types (P < 0.01), the ranges of scar height in the four scar types largely overlapped when judged in a subjective manner, especially in proliferation and superficial scars. CONCLUSIONS: Scar atrophy occurs after one year in burns, but not in the surgical suture wound group. It is important to take anti-scar therapy for surgical wounds as well as burn wounds. The various scar types could not be judged precisely in a subjective manner. PMID- 19187587 TI - High sensitive C-reactive protein, adiponectin, and urine albumin excretion rate in Chinese coronary artery disease patients with different glucose tolerance status. AB - BACKGROUND: Serum high sensitive C-reactive protein (hs-CRP), adiponectin levels and urine albumin excretion rate (UAER) are probably associated with inflammation and atherosclerosis. The aim of this study was to determine the three markers in coronary artery disease (CAD) subjects with different glucose tolerance status in a Chinese population and further explore the levels of the three markers in these subjects and the possible association of these markers with CAD risk factors and the severity of CAD as well. METHODS: A total of 242 subjects with angiographically documented CAD were recruited, and then assigned to three groups: the normal glucose tolerance (NGT) + CAD group, including 100 CAD patients with NGT; the impaired glucose tolerance (IGT) + CAD group, 40 CAD patients with IGT; the type 2 diabetes mellitus (T2DM) + CAD group, 102 CAD patients with T2DM. Serum hs-CRP, adiponectin levels as well as UAER were measured in all subjects. RESULTS: Serum hs-CRP levels were increased in the T2DM + CAD group compared with the NGT + CAD group (4.71 +/- 2.59) vs (3.60 +/- 2.46) mg/L, P = 0.037. Serum adiponectin levels were gradually decreased from the NGT + CAD to IGT + CAD to T2DM + CAD groups, (5.99 +/- 1.84), (5.82 +/- 1.72) and (4.65 +/- 1.71) mg/L, P = 0.002 and 0.040 for NGT + CAD and IGT + CAD groups vs T2DM + CAD group, respectively. While the UAER was gradually increased from the NGT + CAD to IGT + CAD to T2DM + CAD groups, (6.42 +/- 2.51), (6.89 +/- 2.94) and (15.03 +/- 4.22) microg/min (P < 0.001) for NGT + CAD and IGT + CAD groups vs T2DM + CAD group. Multiple linear stepwise regression analysis showed that waist hip ratio (WHR) and low density lipoprotein cholesterol (LDL-C) were the significant determinants of serum hs-CRP levels; triglyceride (TG), high density lipoprotein cholesterol (HDL-C), age, WHR, T2DM, 2-hour serum insulin (2hINS), sex, and apolipoprotein B were the significant determinants of serum adiponectin levels; and systolic blood pressure (SBP), T2DM, and hemoglobin A1c (HbA1c) were the significant determinants of UAER in all subjects (R(2) = 0.070, 0.352, and 0.214, respectively). However, no significant correlation was seen for hs-CRP, adiponectin and UAER with the severity of CAD. Hs-CRP levels were significantly correlated with UAER. CONCLUSIONS: There was a trend of increased serum hs-CRP levels from the NGT + CAD to IGT + CAD to T2DM + CAD groups, though it only showed significance in the T2DM + CAD group compared with the NGT + CAD group. Serum adiponectin levels were decreased and UAER was increased from the NGT + CAD to IGT + CAD to T2DM + CAD groups. Increased UAER and serum hs-CRP, and decreased adiponectin levels were associated with traditional CAD risk factors but failed to be correlated with the severity of CAD. Hs-CRP levels were significantly correlated with UAER. PMID- 19187589 TI - Combined application of virtual imaging techniques and three-dimensional computed tomographic angiography in diagnosing intracranial aneurysms. AB - BACKGROUND: The diagnostic value of virtual imaging combined with three dimensional computed tomographic angiography (3D-CTA) for intracranial aneurysms has not been fully elucidated yet. This study aimed to evaluate the value of combined application of virtual imaging techniques and 3D-CTA in diagnosing patients with aneurismal subarachnoid hemorrhage (SAH) at the acute stage. METHODS: Eighty patients with non-traumatic SAH received 3D-CTA examinations. The raw CT data of these patients were reconstructed and transferred into the 3D mode through the surgical plan system based on virtual reality (VR) image, and the 3D virtual images of skulls and brain blood vessels were acquired. The location, size and shape of aneurysms and their anatomic relationship with adjacent tissues were measured from many points of view. RESULTS: Seventy-three aneurysms were detected in 68 of the 80 patients, but 2 aneurysms were detected in 2 of the 5 patients who had been found free of aneurysms previously and had received 3D-CTA examinations for a second time one month later. The 3D virtual images produced by the virtual imaging system were clear and vivid, and they could reveal the location and size of the aneurysm and its relations to the parent artery and skull directly. CONCLUSIONS: The imaging of 3D-CTA is convenient, reliable and fast in diagnosing intracranial aneurysms and can be regarded as the first choice for the diagnosis and treatment of ruptured intracranial aneurysms. Combined with the surgical plan system based on the VR image, 3D-CTA may obtain more imaging information about aneurysms. PMID- 19187590 TI - Genetic variants in three genes and smoking show strong associations with susceptibility to exudative age-related macular degeneration in a Chinese population. AB - BACKGROUND: The present study was undertaken to replicate the associations of representative polymorphisms in three genes (complement factor H (CFH), complement factor B (BF) and HtrA serine peptidase 1 (HTRA1)) with exudative age related macular degeneration (AMD) in a Han Chinese population, and to test if the modifiable environmental factors affect AMD susceptibility associated with different type of genotype in these genes. METHODS: An age, gender and ethnicity matched case-control study was conducted to genotype the representative single neucleotide polymorphisms (SNPs) loci including rs1061170 and rs1410996 in CFH, rs641153 and rs4151667 in BF and rs11200638 in HTRA1 gene in 144 exudative AMD patients and 126 normal controls using PCR-RFLP and direct resequencing. The demographic characteristics and behavioral risk factors were also recorded. Allelic and genotypic associations for individual SNP and joint associations with two loci were performed. The gene-gene and gene-environment interactions were analyzed using multivariate non-conditional Logistic regression analysis. RESULTS: The C risk allele frequencies for CFH Y402H (rs1061170) in cases and controls were 12.5% and 5.4% respectively, which were much lower than those in Caucasians (P < 0.001). Compared with TT homozygous genotype, the CT heterozygous genotype was positively associated with AMD with odds ratio (OR) of 3.23 (1.36 - 5.07). However, the population attributable risk (PAR) of C allele was only 3.3% (1.4% - 4.3%). rs1410996 was also associated with AMD independent of Y402H. The ORs of exudative AMD for individuals carrying one copy risk allele and two copy risk alleles were 2.57 (1.21 - 5.45) and 4.76 (2.15 - 10.55) respectively, with correspondent PARs of 28.3% (2.0% - 40.5%) and 38.2% (21.8% - 45.4%). rs11200638 in HTRA1 was another susceptible locus for AMD and the risk homozygotes were significantly susceptible for exudutive AMD (OR = 3.98, 1.88 - 8.43) with PAR of 38.9% (24.3% - 45.8%). Education status and cigarette smoking were also related to exudative AMD. After controlling for environmental risk factors, CFH and HTRA1 SNPs were independently associated with exudative AMD, with OR of 3.50 (1.45 - 8.45) for CT genotype in Y402H, 3.34 (1.33 - 8.36) for GG genotype in rs1410996 and 3.85 (1.58 - 9.42) for AA genotype in rs11200638 respectively. The interaction analysis between gene and environmental factors showed that smoking synergistically increased susceptibility of AMD for heterozygotes of rs1410996, with OR(interaction) of 7.33 (P(interaction) = 0.029). CONCLUSIONS: In a Han Chinese population, CFH and HTRA1 polymorphisms appear to be independently and possibly additively hereditary contributors to exudative AMD. Y402H polymorphism conferred a significant but relatively lower contribution in Chinese than in Caucasians with a low frequency of risk allele. The gene-environment interaction may be a best way to encourage those with a high genetic risk to prevent AMD by avoiding modifiable factors until there is effective treatment for AMD. PMID- 19187591 TI - Role of silencing phosphatase of regenerationg liver-3 expression by microRNA interference in the growth of gastric cancer. AB - BACKGROUND: Accumulated evidences demonstrate that phosphatase of regeneration liver-3 (PRL-3) is associated with metastasis of multiple tumor types, and has been validated as a potential therapeutic target for metastasis. High expression of PRL-3 was implicated in lymph node metastasis of gastric cancer. In the present study, we investigated the role of silencing PRL-3 expression by microRNA (miRNA) interference in gastric cancer growth. METHODS: RNA interference, mediated by recombinant lentivirus expressing artificial PRL-3 miRNA, was employed to knockdown PRL-3 expression in human SGC7901 gastric cancer cells. MTT assay and tumor implantation experiment were conducted to determine the role of PRL-3 in the proliferation of SGC7901 cells and tumor growth. RESULTS: Transfection of recombinant lentivirus expressing artificial PRL-3 miRNA significantly suppressed the proliferation of SGC7901 cells in vitro. The implanted tumor size of the PRL-3 transfection group was (1.92 +/- 0.18) cm(3), significantly smaller compared with controls (P < 0.001). CONCLUSIONS: Knockdown of PRL-3 significantly suppressed the proliferation of SGC7901 cells in vitro and tumor growth in vivo. PRL-3 plays a key role in the growth of gastric cancer. PRL 3 should be considered as a potential therapeutic target. PMID- 19187592 TI - Thoracoscopic management for bronchiectasis with non-tuberculous mycobacterial infection. AB - BACKGROUND: Non-tuberculous mycobacteria (NTM) have emerged as important opportunistic pathogens of the human being in recent years. Patients with pre existing bronchiectasis are susceptible to NTM. However, information about its occurrence among bronchiectatic patients in Shenzhen, China is lacking and its impact on the course of bronchiectasis following surgical intervention is unknown. This preliminary study aimed to investigate the prevalence of NTM in bronchiectasis that required surgery in our center, evaluate the role of intraoperative routine screening for NTM, and summarize our initial experience in thoracoscopic management for bronchiectatic patients with NTM. METHODS: A retrospective analysis of clinical, microbiological data of our bronchiectatic patients with NTM over 5 years was made and 40 patients with bronchiectasis were studied to determine the role of intraoperative routine screening for NTM. RESULTS: The prevalence of NTM in this population of patients with bronchiectasis in our center was 6.7% (7/105). The diagnostic yield of the 40 intraoperative specimens was 7.5% (3/40). Of the 7 patients with bronchiectasis and NTM, 3 patients developed postoperative wound infections. All were cured with chemotherapy for 8 - 12 months along with vigorous surgical debridement. Another patient had a slow growth of mycobacteria involving double lungs and the right thoracic cavity and recovered after chemotherapy for nearly 14 months and tube drainage. The affected tissue was completely resected in the remaining 3 patients with no operative mortality and postoperative morbidity, and routine intraoperative screening for NTM was initiated in these patients. CONCLUSIONS: NTM is not uncommon in bronchiectatic patients which deserves surgeons' utmost attention. Routine intraoperative screening for NTM identified otherwise unsuspected patients has shown favorable outcomes. Thoracoscopic management for bronchiectasis with NTM is technically feasible although its role remains to be defined. PMID- 19187593 TI - A novel approach of proteomics to study the mechanism of action of grape seed proanthocyanidin extracts on diabetic retinopathy in rats. AB - BACKGROUND: Diabetic retinopathy (DR) is a leading cause of visual impairment and blindness among the people of occupational age. To prevent the progress of retina injury, effective therapies directed toward the key molecular target are required. Grape seed proanthocyanidin extracts (GSPE) have been reported to be effective in treating diabetic complications, while little is discussed about the functional protein changes. METHODS: We used streptozotocin (STZ) to induce diabetes in rats. GSPE (250 mg/kg body weight per day) were administrated to diabetic rats for 24 weeks. Serum glucose, glycated hemoglobin and advanced glycation end products (AGEs) were determined. Consequently, 2-D difference gel electrophoresis and mass spectrometry were used to investigate retina protein profiles among control, STZ-induced diabetic rats, and GSPE treated diabetic rats. RESULTS: GSPE significantly reduced the AGEs of diabetic rats (P < 0.05). Moreover, GSPE significantly suppressed the vascular lesions of central regions, decreased capillary enlargements and neovascularization, similar to those of the control rats under light microscope. Eighteen proteins were found either up regulated or down-regulated in the retina of STZ-induced diabetic rats. And seven proteins in the retina of diabetic rats were found to be back-regulated to normal levels after GSPE therapy. These back-regulated proteins are involved in many important biological processes such as heat shock, ubiquitin-proteasome system, cell proliferation, cell growth and glucose metabolism. CONCLUSIONS: These findings might promote a better understanding for the mechanism of DR, and provide novel targets for evaluating the effects of GSPE therapy. PMID- 19187594 TI - Effects of carbon disulfide on the expression and activity of nitric oxide synthase in rat hippocampus. AB - BACKGROUND: Carbon disulfide (CS(2)) is a commonly used organic solvent. Many epidemiological investigations and animal experiments have indicated that learning and memory ability can be affected to different degrees after long-term exposure to CS(2), but the mechanisms are still unclear. The aim of this study was to explore the possible mechanisms of CS(2)-related impairment of the learning and memory ability of rats, by investigating the effects of CS(2) on nitric oxide synthase (NOS) activity and NOS mRNA expression in rat hippocampus. METHODS: Rat models of toxicity were generated by inhalation of various doses of CS(2). After two months of inhaling intoxication, the activities of constitutive NOS (cNOS) and induced NOS (iNOS) in the hippocampus were measured. The levels of neuronal NOS (nNOS) mRNA and iNOS mRNA were measured by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). RESULTS: cNOS activity was significantly decreased compared with controls, while iNOS activity was changed only slightly. CS(2) treatment significantly decreased nNOS mRNA levels. iNOS mRNA levels were significantly increased only at higher doses of CS(2). CONCLUSION: The effect of CS2 on learning and memory ability in rats is related to the activity of NOS and the expression of nNOS in the hippocampus. PMID- 19187595 TI - Pathogenicity of Trichosporon asahii in a murine model of disseminated trichosporonosis. AB - BACKGROUND: In recent years, superficial and deep mycoses caused by trichosporon were occasionally reported. In 2001, we reported the first case of disseminated trichosporonosis caused by Trichosporon asahii (T. asahii) in China. In this study, the pathogenicity of T. asahii was investigated in a murine model of disseminated trichosporonosis. METHODS: Seventy-five mice were randomly divided into 7 groups. Each group was inoculated with T. asahii, through intradermal, gastrointestinal tract or intravenous injection. The mice in the experimental groups were given an intraperitoneal injection of cyclophosphamide (CY) to induce granulocytopenia. Mice in the therapeutic group were given both liposomal amphotericin B and fluconazole. The main viscera of the mice were examined by means of tissue culture and pathologic sections. RESULTS: In the two intravenous inoculation groups, T. asahii was isolated from at least one organ in 10 of the 12 granulocytopenic mice and 2 of the 14 immunocompetent mice. Two of the 7 mice in the granulocytopenia group presented with lesions in the inoculation position, but none of the 30 mice in the granulocytopenia and the control group which were inoculated intradermally or through the gastrointestinal tract had viscera infection. In the therapeutic group, the ratio of consequently dead mice, the number of involved viscera, and the incidence of systemic infection were significantly less than the untreated group. Acute purulent inflammation and granulomatous inflammation were the main pathological changes in the course of the infection. Arthrospores and filaments were found in the focus. CONCLUSIONS: T. asahii is an opportunistic pathogen that causes cutaneous and visceral infections in immunologically impaired hosts. An immunocompetent host was to be infected by the invading T. asahii. Several organs, namely the liver, lungs, kidneys, spleen and heart, were predisposed. The therapy of combining liposomal amphotericin B with fluconazole can prevent the host from an infection and inhibit the diffusion of the infection. PMID- 19187596 TI - Effects of activated protein C on coagulation and fibrinolysis in rabbits with endotoxin induced acute lung injury. AB - BACKGROUND: Sepsis induced acute lung injury (ALI) as a common syndrome in clinical practice has a high mortality. Recombinant human activated protein C (APC) can significantly reduce the mortality of patients with severe sepsis. Several studies have implicated that APC may be protective in ALI. METHODS: Twenty-one rabbits were operatively prepared and randomly divided into sham, control, or APC groups (n = 7 in each group). After a tracheotomy had been performed, ALI was produced in the control and APC groups by infusion of Escherichia coli endotoxin 100 microg/kg per hour intravenously for 1 hour. The sham group received only the vehicle, infusion of 20 ml of 0.9% saline. The rabbits were studied under anesthesia for 6 hours and were ventilated with 40% oxygen. Bovine APC (25 microg x kg(-1) x h(-1)) was intravenously administered. The infusion was initiated half an hour post-injury and lasted for 4 hours. The animals were resuscitated with Ringer's lactate solution. RESULTS: In comparison with nontreatment in the control group, the infusion of APC significantly reduced the increase of thrombomodulin level (TM; control group was (0.68 +/- 0.06) ng/ml, vs APC group of (0.62 +/- 0.07) ng/ml at 6 hours, P < 0.05), and significantly attenuated the fall in protein S (PS; control group was (2.32 +/- 0.03) microg/ml at 2 hours, (2.24 +/- 0.06) microg/ml at 4 hours and (2.21 +/- 0.09) microg/ml at 6 hours, vs APC group (2.46 +/- 0.04) microg/ml at 2 hours, (2.40 +/- 0.05) microg/ml at 4 hours and (2.39 +/- 0.07) microg/ml at 6 hours, P < 0.01). In addition, APC limited the increase in plasminogen activator inhibitor 1 (PAI-1) both in plasma (control group was (0.68 +/- 0.12) ng/ml at 1 hour, (0.84 +/- 0.06) ng/ml at 2 hours, (0.87 +/- 0.08) ng/ml at 4 hours and (0.91 +/- 0.05) ng/ml at 6 hours, vs APC group (0.42 +/- 0.16) ng/ml at 1 hour, (0.43 +/- 0.04) ng/ml at 2 hours, (0.45 +/- 0.09) ng/ml at 4 hours and (0.45 +/- 0.14) ng/ml at 6 hours, P < 0.01) and in bronchoalveolar lavage fluid (at 6 hours: sham, (1.05 +/- 0.05) ng/ml; control, (1.13 +/- 0.06) ng/ml; APC, (1.06 +/- 0.06) ng/ml; P < 0.05). However, APC failed to prevent the decrease in PaO(2)/FiO(2) ratio. APC-treated rabbits showed no significant difference in platelet count and antithrombin but exhibited less D-dimer production than did the controls. Moreover, APC limited the histopathological score of lung injury (2.6 +/- 0.8 in control, vs 1.4 +/- 0.6 in APC group, P < 0.01). CONCLUSION: Anti-coagulation and pro-fibrinolysis activity may be two of the possible mechanisms by which activated protein C attenuated endotoxin-induced ALI. PMID- 19187597 TI - Caspase-dependent retinal ganglion cell apoptosis in the rat model of acute diabetes. AB - BACKGROUND: Neural apoptosis is generally believed to be mediated by two distinct pathways, caspase-dependant and caspase-independent pathways. This study investigated the apoptotic pathways involved in retinal ganglion cells in acute diabetes in rats. METHODS: Diabetes was induced in male Wistar rats by a peritoneal injection of streptozotocin (STZ). Expression and localization of caspase-3 and apoptosis-inducing factor (AIF) proteins in the retina of diabetic rats was examined by Western blotting and immunohistochemistry analyses. Terminal transferase dUTP nick end labeling (TUNEL) assay and immunofluorescent staining specific for caspase-3 and AIF were applied to analyze for apoptosis of retinal ganglion cells. In addition, a caspase-3 inhibitor DEVD-CHO was injected intravitreally to further determine the apoptotic pathways of retinal ganglion cells triggered in acute diabetes. RESULTS: Two weeks after induction of diabetes, a significant increase in caspase-3 protein expression and localization occurred in the nerve fiber layer, ganglion cell layer, and inner plexiform layer of the retina. Four weeks after the onset of diabetes, the increase in caspase-3 expression was profound eight weeks postinduction of diabetes (P < 0.05). Meanwhile, no AIF protein expression was detected in this study. In addition, intravitreal administration of the caspase-3 inhibitor DEVD-CHO reduced apoptosis of retinal ganglion cells by its direct inhibitory action on caspase-3. CONCLUSION: Caspase-dependent apoptotic pathways may be the main stimulant of STZ induced retinal ganglion cell apoptosis in acute diabetes. PMID- 19187598 TI - Therapeutic time window of flurbiprofen axetil's neuroprotective effect in a rat model of transient focal cerebral ischemia. AB - BACKGROUND: The neuroprotective effect of the cyclooxygenase (COX) inhibitor has been demonstrated in acute and chronic neurodegenerative processes. But its function under cerebral ischemic conditions is unclear. This study was designed to evaluate the neuroprotective efficacy of emulsified flurbiprofen axetil (FA, COX inhibitor) and its therapeutic time window in a model of transient middle cerebral artery occlusion (MCAO) in rats. METHODS: Forty-eight male SD rats were randomly assigned into six groups (n = 8 in each group); three FA groups, vehicle, sham and ischemia/reperfusion (I/R) groups. Three doses of FA (5, 10 or 20 mg/kg, intravenous infusion) were administered just after cerebral ischemia/reperfusion (I/R). The degree of neurological outcome was measured by the neurologic deficit score (NDS) at 24, 48 and 72 hours after I/R. Mean brain infarct volume percentage (MBIVP) was determined with 2, 3, 5 triphenyltetrazolium chloride (TTC) staining at 72 hours after I/R. In three other groups (n = 8 in each group), the selected dosage of 10 mg/kg was administrated intravenously at 6, 12 and 24 hours after I/R. RESULTS: The three different doses of FA improved NDS at 24, 48 and 72 hours after I/R and significantly reduced MBIVP. However, the degree of MBIVP in the FA 20 mg/kg group differed from that in FA 10 mg/kg group. Of interest is the finding that the neuroprotective effect conferred by 10 mg/kg of FA was also observed when treatment was delayed until 12 - 24 hours after ischemia reperfusion. CONCLUSION: COX inhibitor FA is a promising therapeutic strategy for cerebral ischemia and its therapeutic time window could last for 12 - 24 hours after cerebral ischemia reperfusion, which would help in lessening the initial ischemic brain damage. PMID- 19187599 TI - Inducible nitric oxide synthase is involved in the oxidation stress induced by HIV-1 gp120 in human retina pigment epithelial cells. AB - BACKGROUND: The human immunodeficiency virus-1 (HIV-1) envelope glycoprotein gp120 has been implicated in the development of AIDS-associated retinopathy. The present study tested the hypothesis that gp120 may induce oxidative stress including up regulation of inducible nitric oxide synthase (iNOS) and production of malondialdehyde (MDA) and nitric oxide (NO) to mediate retinopathy in retinal pigment epithelial (RPE) cells. METHODS: Human RPE cell line D407 was cultured and treated with gp120. HIV-1 gp120 protein induced lipid peroxidation product MDA. NO production and iNOS expression were examined in vitro by spectrophomtometry, real-time PCR, Western blotting, and confocal microscope. RESULTS: Addition of gp120 was able to induce RPE cells to produce NO and MDA in time- and dose-dependent manners (P < 0.05). Similarly, gp120 was also capable of up-regulating iNOS mRNA and protein in D407 cells in time- and dose-dependent manners. CONCLUSIONS: Gp120 induces oxidative stress in D407 cell by stimulating MDA and NO production, which is mediated by up-regulating iNOS expression. Gp120 may mediate oxidation stress in AIDS-associated retinopathy. PMID- 19187600 TI - High extracellular potassium ion concentration attenuates the blockade action of ketanserin on Kv1.3 channels expressed in xenopus oocytes. AB - BACKGROUND: Ketanserin (KT), a selective serotonin (5-HT) 2-receptor antagonist, reduces peripheral blood pressure by blocking the activation of peripheral 5-HT receptors. In this study electrophysiological method was used to investigate the effect of KT and potassium ion on Kv1.3 potassium channels and explore the role of blocker KT in the alteration of channel kinetics contributing to the potassium ion imbalances. METHODS: Kv1.3 channels were expressed in xenopus oocytes, and currents were measured using the two-microelectrode voltage-clamp technique. RESULTS: KCl made a left shift of activation and an inactivation curve of Kv1.3 current and accelerated the activation and inactivation time constant. High extracellular [K(+)] attenuated the blockade effect of KT on Kv1.3 channels. In the presence of KT and KCl the activation and inactivation time constants were not influenced significantly no matter what was administered first. KT did not significantly inhibit Kv1.3 current induced by tetraethylammonium (TEA). CONCLUSIONS: KT is a weak blocker of Kv1.3 channels at different concentrations of extracellular potassium and binds to the intracellular side of the channel pore. The inhibitor KT of ion channels is not fully effective in clinical use because of high [K(+)](o) and other electrolyte disorders. PMID- 19187601 TI - Cardiotrophin-1 induces intercellular adhesion molecule-1 expression by nuclear factor kappaB activation in human umbilical vein endothelial cells. AB - BACKGROUND: In addition to elevated concentrations of cytokines, patients with congestive heart failure (CHF) show endothelial dysfunction and increased plasma concentrations of adhesion molecules like intercellular adhesion molecule-1 (ICAM 1). Furthermore, the concentration of cardiotrophin-1 (CT-1)--a cytokine of the interleukin-6 superfamily--is increased in CHF. We tested the hypothesis whether CT-1 is able to induce ICAM-1 in human umbilical vein endothelial cells (HUVEC). Furthermore we examined the signalling mechanisms of CT-1 mediated ICAM-1 expression. METHODS: Confluent layers of HUVEC were incubated with increasing concentrations of CT-1 (5 to 100 ng/ml) for different periods. ICAM-1 mRNA was determined by real-time polymerase chain reaction (PCR) and ICAM-1 surface expression by fluorescence-activated cell sorter (FACS) analysis and soluble ICAM 1 (sICAM-1) in the culture supernatant by enzyme linked immunosorbent assay (ELISA). To clarify the signalling pathway of CT-1 induced ICAM-1 expression we used various inhibitors of possible signal transducing molecules, electromobility shift assay (EMSA) and Western blot analysis. RESULTS: CT-1 induced ICAM-1 mRNA (1.8 +/- 0.8 fold increase compared to unstimulated cells after 6 hours) and protein (1.4 +/- 0.2 fold increase compared to unstimulated cells after 48 hours) in HUVEC in a time- and concentration-dependent manner. EMSA experiments show that CT-1 causes nuclear factor (NF) kappaB activation. Because parthenolide could inhibit CT-1 induced ICAM-1 expression NFkappaB activation is required in this pathway. CT-1 did not activate extracellular signal regulated kinases (ERK), c-Jun N-terminal kinase (JNK) and p38. CONCLUSION: CT-1 is able to induce ICAM-1 in endothelial cells by NFkappaB activation. These results may explain in part elevated ICAM-1 concentrations in patients with CHF and endothelial dysfunction. PMID- 19187602 TI - Roles of cyclooxygenase-2 in microvascular endothelial cell proliferation induced by basic fibroblast growth factor. AB - BACKGROUND: The level of basic fibroblast growth factor (bFGF) increases rapidly after cerebral ischemia. However, the molecular mechanisms for the effects of bFGF on cerebral microvascular endothelial cells (cMVECs) have not yet been fully elucidated. In this study, a murine cMVEC line, bEnd.3, was employed to study the effects of bFGF on cyclooxygenase (COX) expression and its downstream effects in cMVECs. METHODS: After treatment with bFGF, RT-PCR and Western blotting analyses were carried out to evaluate the changes in COX-2 mRNA and protein expression, respectively. MTT assays were performed to measure cell proliferation. The prostaglandin E2 (PGE2) and vascular endothelial growth factor (VEGF) concentrations in the culture medium were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: COX-2 mRNA and protein expressions in bEnd.3 cells were induced by bFGF in time- and dose-dependent manners. The bFGF-induced COX-2 upregulation led to enhanced PGE2 production by bEnd.3 cells, and this effect was abolished by the selective COX-2 inhibitor NS-398. bFGF also increased VEGF production by bEnd.3 cells, and this effect was blocked by NS-398 and the EP1/2 (PGE2 receptors) antagonist AH6809. Furthermore, exogenous PGE2 increased VEGF production in bEnd.3 cells, and AH6809 blocked this effect. CONCLUSION: bFGF increases VEGF production in an autocrine manner by increasing COX-2-generated PGE2 in cMVECs and subsequently stimulates MVEC proliferation and angiogenesis. PMID- 19187603 TI - Mechanisms responsible for pulmonary hypertension. PMID- 19187604 TI - Biliary papillomatosis: analysis of 18 cases. PMID- 19187605 TI - Factors influencing the natural history of HIV-1 infection. PMID- 19187606 TI - Extracorporeal membrane oxygenation for pulmonary hemorrhage in microscopic polyangiitis. PMID- 19187607 TI - Prenatal diagnosis of fetal aortopulmonary septal defect with ventricular septal defect by two-dimension echocardiography. PMID- 19187608 TI - Management of hepatitis B in China. PMID- 19187609 TI - Prevalence of antinuclear and anti-liver-kidney-microsome type-1 antibodies in patients with chronic hepatitis C in China. AB - BACKGROUND: Hepatitis C virus (HCV) infection may induce autoimmune response and autoantibodies can be detected in chronic hepatitis C (CHC) patients. However, the reported positive rate of autoantibodies in CHC patients in China varies considerably. In this study, we investigated the prevalence of antinuclear antibodies (ANA) and anti-liver-kidney-microsome type 1 autoantibodies (anti-LKM 1) in a large cohort of CHC patients, and analyzed the factors related to the presence of the autoantibodies. METHODS: A total of 360 CHC patients were enrolled in this study. Serum ANA and anti-LKM-1 were detected by indirect immunofluorescence and enzyme-linked immunosorbent assay, respectively. Clinical analysis was performed to disclose the related factors to autoantibody production. RESULTS: The prevalence of ANA and anti-LKM-1 in CHC patients was 12.5% (45/360) and 2.5% (9/360), respectively. Women had a higher prevalence than men (18.9% vs 11.4%, P = 0.046). Patients with positive autoantibodies had lower HCV RNA levels (1.2 x 10(7) copies/L vs 7.2 x 10(7) copies/L, P < 0.05). Positive ANA was associated with higher serum globulin (P < 0.05). Stratified analysis showed that there were no significant differences in age, HCV genotype, disease course, clinical stage, prevalence of cirrhosis and interferon therapy between autoantibody-positive and -negative subgroups. CONCLUSION: Autoantibodies can be induced in the course of CHC, and some CHC patients can even develop autoimmune hepatitis. PMID- 19187610 TI - Altered expression profiles of microRNAs in a stable hepatitis B virus-expressing cell line. AB - BACKGROUND: MicroRNAs (miRNAs) are highly conserved small non-coding RNAs of 18 - 25 nucleotides (nt) that mediate post-transcriptional gene regulation. Hepatitis B virus (HBV) can cause either acute or chronic hepatitis B, and is a high risk factor for liver cirrhosis and hepatocellular carcinoma. Some mammalian viruses have been shown to modulate the expression of host cellular miRNAs. However, interactions between the HBV and the host cellular miRNAs are largely unknown. METHODS: miRNA microarray and Northern blotting analysis were used to compare the expression profile of cellular miRNAs of a stable HBV-expressing cell line HepG2.2.15 and its parent cell line HepG2. mRNA microarray assay and the miRanda program were used to predict the miRNA targets. A flow cytometric assay was further used to investigate the expression of human leukocyte antigen (HLA)-A. RESULTS: Eighteen miRNAs were differentially expressed between the two cell lines. Among them, eleven were up-regulated and seven were down-regulated in HepG2.2.15 cells. Northern blotting analysis confirmed that the expression of miR 181a, miR-181b, miR-200b and miR-146a were up-regulated and the expression of miR 15a was down-regulated, which was in consistent with the results of the microarray analysis. Furthermore, some putative miRNA targets were predicted and verified to be linked with mRNA expression. The 3'-UTR of HLA-A gene had one partially complementary site for miR-181a and miR-181a might down-regulate the expression of HLA-A. CONCLUSION: HBV replication modulates the expression of host cellular miRNAs, which may play a role in the pathogenesis of HBV-related liver diseases. PMID- 19187611 TI - Proteomic analysis of differently expressed proteins in human hepatocellular carcinoma cell lines HepG2 with transfecting hepatitis B virus X gene. AB - BACKGROUND: Hepatitis B virus encoded X protein (HBx) is a trans-activating protein that may be involved in hepatocarcinogenesis, although few natural effectors of HBx that participate in this process have been identified. We screened, by comparative proteomics method, effectors of HBx associated with hepatocarcinogenesis. METHODS: HBx positive and negative HepG2 cells were constructed and expression patterns of cellular proteins were obtained by high resolution, two dimensional electrophoresis. Comprehensive analyses of proteins associated with hepatocellular carcinoma (HCC) were focused on the differently expressed proteins (more than two-fold increase or decrease, P < 0.05) from HBx positive and negative HepG2 cells. For peptide mass fingerprinting, protein spots with different intensity between HBx positive and negative HepG2 cells were directly cut out of gels and processed for matrix assisted, laser desorption/ionization, time of flight mass spectrometry and liquid chromatography tandem mass spectrometry (LC-ESI-MS/MS) analysis. RESULTS: The mean number of protein spots for HBx negative and HBx positive HepG2 cells were 2095 +/- 137 and 2188 +/- 105, respectively. The analysis of paired cells showed 75 spots with significant differences in expression between HBx negative and HBx positive cells: 37 spots corresponding to 32 different proteins; 25 proteins were upregulated, 7 downregulated. We found 7 proteins not previously reported differentially expressed in HBx positive HepG2 cells. Variations in protein accumulation were confirmed for four (HSP90AB1, BCL2 associated athanogene 2, nucleophosmin and chloride intracellular channel 1) by Western blotting in HBx positive HepG2 cells. CONCLUSIONS: Numerous effectors of HBx that may promote the development of HCC are identified, of which 7 are newly noted in HepG2 cells. Several of these effectors of HBx may help in elucidating the roles of HBx in hepatocarcinogenesis and diagnostics or targets for therapeutic intervention. PMID- 19187612 TI - Association among plasma interleukin-18 levels, carotid intima- media thickness and severity of obstructive sleep apnea. AB - BACKGROUND: Epidermic studies have suggested a pathophysiological link between obstructive sleep apnea hypopnea syndrome (OSAHS) and atherosclerosis (AS); for which carotid intima-media thickness (IMT) has been considered as an early marker. The pathogenesis by which OSAHS can induce AS has not been elucidated. This study was conducted to investigate the association among plasma interleukin 18 (IL-18) levels, carotid IMT and the severity of OSAHS. METHODS: Based on the apnea hypopnea index (AHI) during sleep monitored by polysomnography, 52 male patients with OSAHS were recruited as the OSAHS group which was further divided into mild OSAHS (n = 16), moderate OSAHS (n = 18), and severe OSAHS (n = 18) subgroups. Eighteen healthy subjects were selected as the control group. Of all OSAHS patients, 20 with moderate-to-severe OSAHS underwent continuous positive airway pressure (CPAP) treatment for 90 days. HDL5000 color Doppler ultrasonography was used to measure carotid IMT. Plasma IL-18 levels were measured by ELISA. RESULTS: Compared with the plasma IL-18 levels in the control group ((250.27 +/- 76.48) pg/ml), there was a significant increase in the mild OSAHS subgroup ((352.08 +/- 76.32) pg/ml), the moderate subgroup ((600.17 +/- 83.91) pg/ml), and the severe OSAHS subgroup ((9797.64 +/- 109.83) pg/ml) (all P < 0.01). Moreover, there was a significant difference in plasma IL-18 levels among the three OSAHS subgroups (P < 0.01). Carotid IMT was significantly greater in the severe OSAHS subgroup than in the mild OSAHS subgroup (P < 0.01). Before CPAP treatment, plasma IL-18 levels were positively correlated with carotid IMT (r = 0.486, P < 0.001) and with AHI (r = 0.865, P < 0.001). On day 90 of CPAP treatment, plasma IL-18 levels were significantly declined but carotid IMT was not changed significantly. CONCLUSIONS: In untreated OSAHS patients carotid IMT and plasma IL-18 were positively correlated and were significantly higher than in normal controls; the elevation of plasma IL-18 levels was correlated with the severity of OSAHS. Inflammatory response associated with OSAHS may be related to the development of AS. By improving AHI, miniSaO(2), and reducing plasma IL-18 levels, CPAP treatment may slow down or prevent the development of AS in OSAHS patients. PMID- 19187613 TI - Association of TBX5 gene polymorphism with ventricular septal defect in the Chinese Han population. AB - BACKGROUND: Congenital heart disease is a diverse group of diseases determined by genetic and environmental factors. Considerable research has been done on genes associated with development of the heart. A recent focus is the role of transcription factor TBX5 in the development of atria, left ventricle and conduction system. As part of a larger study, high density, single nucleotide polymorphism (SNP) scanning was used to explore the relationship between TBX5 gene polymorphism and susceptibility to ventricular septal defect not associated with forelimb malformation in the Chinese Han population. METHODS: One hundred and ninety two paediatric patients with congenital ventricular septal defect and 192 matched healthy control subjects were studied. The haplotype reconstructions were calculated by PHASE2.0 software. Haploview software was used to perform linkage disequilibrium assessment and defining of haplotype blocks. The algorithm used for defining of blocks was the confidence interval method. RESULTS: The TBX5 gene region can be divided into 3 haplotype blocks of 27, 15 and 2 SNPs. Strong linkage disequilibrium exists within each block. SNP rs11067075 within the TBX5 gene had significant correlation with ventricular septal defect (P = 0.0037) by single marker association analysis. In addition, a 20 kb haplotype composed of 27 SNPs correlated with ventricular septal defect (P = 0.05, multiple loci regression analyses based on reconstructed haplotype blocks). CONCLUSIONS: TBX5 is associated with the occurrence of ventricular septal defect and may be a predisposing gene to congenital heart disease in Han Chinese. This finding has set a direction for further genetic and functional studies. PMID- 19187614 TI - Rapamycin instead of mycophenolate mofetil or azathioprine in treatment of post renal transplantation urothelial carcinoma. AB - BACKGROUND: Malignant tumor is the most common complication occurred in transplant recipients. It is widely recognized that immunosuppressive treatments increase the risk of cancer in transplant recipients. The efficacy and safety of rapamycin (RPM) in combination with low-dose calcineurin inhibitor (CNI) in treating 15 renal allograft recipients which developed urothelial carcinoma were observed. METHODS: Immunosuppressive regimen in all recipients was altered with rapamycin to replace mycophenolate mofetil (MMF) or azathioprine (Aza). The initial loading dosage was 2 mg/d, and the next dosage was 1 mg/d. The dosage of rapamycin was carefully adjusted according to the blood drug level and concentration of the drug was maintained at 4 - 6 microg/L. In all the 15 patients, the calcineurin inhibitor was reduced down to one third of the original dosage after the rapamycin blood concentration became stable. Surgical treatment and intravesical instillation chemotherapy were carried out in all patients. Recurrence of the tumor was monitored throughout the study. Post-transplant renal function and side effects were also closely monitored. RESULTS: Among the 15 patients, 9 had no tumor recurrence in 2 years, 2 had tumor recurrences twice, and 4 had once. There was no acute rejection observed during RPM treatment. Post transplant renal function in 11 patients was improved, with a decreased creatinine level. Hyperlipoidemia and thrombocytopenia were the most frequent adverse events which responded well to corresponding treatments. CONCLUSION: Among the renal allograft recipients with urothelial carcinoma, combination of rapamycin and low dose calcineurin inhibitor treatment is effective and safe. PMID- 19187615 TI - Preliminary study of 3T 1H MR spectroscopy in bone and soft tissue tumors. AB - BACKGROUND: Magnetic resonance spectroscopy (MRS) is one method that can examine noninvasively the alive specimen of the organ, metabolism of the organ and cell, and the biochemistry change. MRS provides the biochemistry information that may be used to diagnose tumors or differentiate the malignant tumor from benign. The objective of this study is to investigate the benign and malignant bone and soft tissue tumors by 1H-MR spectroscopy ((1)H-MRS) on a 3 Tesla MR scanner, then to assess the usefulness of (1)H-MRS in diagnosing bone and soft tissue tumors and distinguishing benign from malignant tumors. METHODS: Fifty-six patients with bone and soft tissue tumors proved clinically and pathologically were examined with (1)H-MRS. (1)H-MRS was performed to study malignant musculoskeletal tumors, benign tumors and normal muscle adjacent to lesions to analyze the characteristics, and single-voxel point-resolved spectroscopy sequence was used. Proton brain exam-single voxel of (1)H-MRS which directly appeared in the spectrum, was observed to find the peak height of choline compounds (Cho) opposite to the creatine (Cr), and whether there was a Cho peak. Metabolite values were calculated automatically from the area under each metabolite peak by the Functool 3.1 software. Metabolite ratios of Cho/Cr were manually calculated. Then according to the results, it was judged whether there existed benign or malignant tumors. The Kappa statistical test was used to analyze the MRS results, the histopathology data and the surgical situation. Statistics processing was performed using the software package SPSS11.5 for Windows. RESULTS: (1)H-MRS spectra style of bone and soft tissue tumors was different from that of normal muscle, and differences also existed between benign and malignant tumors. Choline level in malignant tumor was markedly higher than that in benign tumors. Cho/Cr in malignant tumor was higher than in benign tumor significantly (P < 0.05). The true positive rate of bone and soft tissue between benign and malignant tumors was 34/36, the true negative rate was 15/18, the false positive rate was 3/18 and the false negative rate was 2/36. Therefore in the group, sensitivity of the (1)H MRS was 94% (34/36), specificity was 83% (15/18), positive predictive value was 92% (34/37), negative predictive value was 88% (15/17) and the accuracy rate was 91% (49/54). The MRS results and the histopathology inspection conclusions had very good uniformity. The kappa value was 0.76 +/- 0.10 (P < 0.01). CONCLUSIONS: The increase of Cho level measured by (1)H-MRS is related to the bone and soft tissue malignant tumor. Cho/Cr in malignant tumor was higher than in benign tumor, so they will play a vital role in the diagnosis and differential diagnosis of bone and soft tissue tumors. PMID- 19187616 TI - Comparison of bolus remifentanil versus bolus fentanyl for blunting cardiovascular intubation responses in children: a randomized, double-blind study. AB - BACKGROUND: The authors found no study to compare the efficacy of bolus dose fentanyl and remifentanil blunting the cardiovascular intubation response in children, so they designed this randomized, double-blind clinical study to assess the effects of remifentanil 2 microg/kg and fentanyl 2 microg/kg by bolus injection on the cardiovascular intubation response in healthy children. METHODS: One hundred and two children, the American Society of Anesthesiologists (ASA) physical status 1-2 and scheduled for elective plastic surgery under general anesthesia, were randomly divided into one of two groups to receive the following treatments in a double blind manner: remifentanil 2 microg/kg (Group R) and fentanyl 2 microg/kg (Group F) when anesthesia was induced with propofol and vecuronium. The orotracheal intubation was performed using a direct laryngoscope. Blood pressure (BP) and heart rate (HR) were recorded before anesthesia induction (baseline values), immediately before intubation, at intubation and every minute for 5 minutes after intubation. The percent changes of systolic blood pressure (SBP) and HR relative to baseline values and the rate pressure product (RPP) at every observing point were calculated. The incidences of SBP and HR percent changes >30% of baseline values and RPP >22,000 during the observation were recorded. RESULTS: There were no significant differences between groups in the demographic data, baseline values of BP and HR and the intubation time. As compared to baseline values, BP, HR and RPP at intubation and their maximum values during observation increased significantly in Group F, but they all decreased significantly in Group R. BP, HR and RPP at all observed points, and their maximum values during the observation, were significantly different between groups. There were also significant differences between groups in the percent change of SBP and HR relative to baseline values at all observed points and their maximum percent changes during the observation. The incidences of SBP and HR percent increased >30% of the baseline values and RPP >22,000 during the observation, were significantly higher in Group F than in Group R, but the incidences of SBP and HR percent decreased >30% of baseline values were significantly lower in Group F compared with Group R. CONCLUSIONS: When used as part of routine anesthesia induction with propofol and vecuronium in children, fentanyl 2 microg/kg by bolus injection fails to effectively depress the cardiovascular intubation response. Remifentanil 2 microg/kg by bolus injection can completely abolish the cardiovascular intubation response, but also cause more adverse complications of temporary significant cardiovascular depression. PMID- 19187617 TI - Analysis of soft tissue sarcomas in 1118 cases. AB - BACKGROUND: It is important to analyze and compare soft tissue sarcomas periodically so as to update the incidence, the clinical diagnosis, the treatment, and the ongoing research. The present study was conducted to determine the relative frequency of each type of soft tissue sarcoma. METHODS: A total of 1118 cases of primary soft tissue sarcomas treated between January 1993 and December 2006 were evaluated in a retrospective analysis. RESULTS: According to the pathologic grouping, the diseases with the highest proportion were malignant fibrous histiocytomas (35.24%), synovial sarcomas (17.08%), liposarcomas (16.28%), and rhabdomyosarcomas (12.61%). Soft tissue sarcomas were detected in every age group and occurred in all parts of the body. The number of cases increased gradually over the years. CONCLUSIONS: Malignant fibrous histiocytomas had the highest frequency among the soft tissue sarcomas. The number of cases increased gradually over the years. PMID- 19187618 TI - Involvement of connexin 43 in acupuncture analgesia. AB - BACKGROUND: Connexin 43 (Cx43) is one of the major components of human keratinocyte gap junctions. To study whether gap junctional intercellular communication participates in the transfer of acupoint signals and acupuncture analgesia, the expression of Cx43 was studied in Zusanli (ST36) acupoints compared with control non-acupoint regions in rats after acupuncture. In addition, Cx43 heterozygous gene knockout mice were used to further explore the relationship between Cx43 and acupuncture analgesia. METHODS: The expression of Cx43 was detected by immunohistochemistry, immunoblotting, and RT-PCR for the Cx43 protein and mRNA. The influence of the Cx43 gene knockout on acupuncture analgesia was measured by a hot plate and observing the writhing response on Cx43 heterozygous gene knockout mice. RESULTS: Immunohistochemistry showed abundant Cx43 expression in some cells in the skin and subcutaneous tissue of rat ST36 acupoints. The mRNA and protein levels of Cx43 in acupoints were significantly higher than those in the control points in the non-acupuncture group, and even more so after acupuncture. The hot plate and writhing response experiments showed that partial knockout of the Cx43 gene decreased acupuncture analgesia. CONCLUSION: Cx43 expression and acupuncture analgesia showed a positive correlation. PMID- 19187619 TI - Dihydroxyflavonol reduces post-infarction left ventricular remodeling by preventing myocyte apoptosis in the non-infarcted zone in goats. AB - BACKGROUND: Myocyte apoptosis is considered to be the major causative factor of left ventricular (LV) remodeling following myocardial infarction (MI). We previously reported that 3', 4'-dihydroxyflavonol (DiOHF), was able to suppress oxidative stress and preserve the expression of endothelial nitric oxide synthase during myocardial reperfusion injury, which may benefit the reduction of myocyte apoptosis. We therefore aimed to evaluate the potential actions of DiOHF against myocyte apoptosis and post-infarction LV remodeling in this study. METHODS: Following experimental MI, surgical instrumented goats were randomly assigned into vehicle and DiOHF (2 mg/kg; i.v., daily) groups to receive 4 weeks of reperfusion with corresponding treatments. LV pressure recordings and echocardiogram were performed at baseline, 2 and 4 weeks of reperfusion. Myocardial tissues were collected in the end to determine infarct size and apoptosis related assays. RESULTS: LV end-diastolic volume and diameter were significantly increased 4 weeks after MI in the vehicle group, accompanied by reduced posterior wall thickness, septal thickness and LV mass, whereas those changes were markedly prevented by DiOHF treatment. Similarly, significantly reduced infarct size was found in DiOHF group as compared to vehicle group, and DiOHF dramatically inhibited the increase in LV end-diastolic pressure and the reductions in ejection fraction, fraction shortening and dP/dt(max). Moreover, DiOHF treatment significantly reduced the extent of myocyte apoptosis detected by TUNEL assay, enhanced the protein expression of caspase-3, Fas, Bax and cytochrome c in the non-infarcted myocardium in comparison to vehicle. CONCLUSIONS: Daily DiOHF treatment during the reperfusion period after MI in the ovine hearts markedly reduced the magnitude of post-infarction LV remodeling through the inhibition of myocyte apoptosis in the remote non-infarcted myocardium. PMID- 19187620 TI - Effect of chronic intermittent hypoxia on the expression of Nip3, cell apoptosis, beta-amyloid protein deposit in mice brain cortex. AB - BACKGROUND: Chronic intermittent hypoxia (CIH) is the most important pathophysiologic feature of sleep apnea syndrome (SAS). To explore the relationship between SAS and dementia, the effects of CIH on the expression of Nip3, neuron apoptosis and beta-amyloid protein deposit in the brain cortex of the frontal lobe of mice were evaluated in this study. METHODS: Thirty male ICR mice were divided into four groups: control group (A, n = 10, sham hypoxia/reoxygenation), 2 weeks CIH group (B, n = 5), 4 weeks CIH group (C, n = 5), and 8 weeks CIH group (D, n = 10). The ICR mice were placed in a chamber and exposed to intermittent hypoxia (oxygen concentration changed periodically from (21.72 +/- 0.55)% to (6.84 +/- 0.47)% every two minutes, eight hours per day). Neuron apoptosis of the cortex of the frontal lobe was detected by means of terminal deoxy-nucleotidyl transferase-mediated in situ end labeling (TUNEL). Immunohistochemical staining was performed for measuring expression of Nip3 and beta-amyloid protein. The ultrastructure of neurons was observed under a transmission electron microscope. RESULTS: TUNEL positive neurons in each square millimeter in the cortex of the frontal lobe were categorized by median or Ri into group A (1, 5.5), group B (133, 13), group C (252, 21), and group D (318, 24). There were significant differences among the above four groups (P = 0.000). The significance test was performed between the control group and each CIH group respectively: group A and B (P > 0.05); group A and C (P < 0.01); and group A and D (P < 0.005). The number of apoptotic neurons kept increasing in the ICR mice under CIH condition, and reached the peak in the group D, but there was no significant difference between groups B and C, between groups B and D, and between groups C and D. Nip3 positive neurons in each square millimeter in the cortex of the frontal lobe in each group were calculated by median or Ri as follows: group A (2, 5.5), group B (117, 13), group C (227, 26.2), and group D (479, 21.4). There were significant differences among the four groups (P = 0.000). The statistical test was performed between the control group and each CIH group respectively: groups A and B (P > 0.05); groups A and C (P < 0.005); and groups A and D (P < 0.005). There was no significant difference between groups B and C, groups B and D, and groups C and D. The expression of Nip3 was closely correlated with neuron apoptosis in the brain (P < 0.05). The expression of beta amyloid protein in the brain of mice was negative in all CIH groups and the control group. Ultrastructure observation showed karyopyknosis of nucleus, swelling of chondriosomes, deposit of lipofuscins and degeneration of neural sheath in all CIH groups but not in the control group. CONCLUSION: The results of this study indicate that CIH could up-regulate the expression of Nip3, and result in neuron apoptosis and ultrastructural changes in neurons of the frontal cortex. PMID- 19187621 TI - Probucol attenuates atrial autonomic remodeling in a canine model of atrial fibrillation produced by prolonged atrial pacing. AB - BACKGROUND: We hypothesize that increased atrial oxidative stress and inflammation may play an important role in atrial nerve sprouting and heterogeneous sympathetic hyperinnervation during atrial fibrillation (AF). To test the hypothesis, we examined whether the antioxidant and anti-inflammatory treatment with probucol attenuates atrial autonomic remodeling in a canine model of AF produced by prolonged rapid right atrial pacing. METHODS: Twenty-one dogs were divided into a sham-operated group, a control group and a probucol group. Dogs in the control group and probucol group underwent right atrial pacing at 400 beats per minute for 6 weeks, and those in the probucol group received probucol 1 week before rapid atrial pacing until pacing stopped. After 6-week rapid atrial pacing, general properties including left atrial structure and function, atrial hemodynamics and the inducibility and duration of AF were measured in all the groups. Atrial oxidative stress markers and serum C-reactive protein (CRP) concentration were estimated. The degree of nerve sprouting and sympathetic innervation at the right atrial anterior wall (RAAW) and the left atrial anterior wall (LAAW) were quantified by immunohistochemistry, atrial norepinephrine contents were also detected. Atrial beta-nerve growth factor (beta-NGF) mRNA and protein expression at the RAAW and LAAW were assessed by real-time quantitative RT-PCR and Western blotting respectively. RESULTS: Atrial tachypacing induced significant nerve sprouting and heterogeneous sympathetic hyperinnervation, and the magnitude of nerve sprouting and hyperinnervation was higher in the RAAW than in the LAAW. Atrial beta-NGF mRNA and protein levels were significantly increased at the RAAW and LAAW, and the upregulation of beta-NGF expression was greater at the RAAW than at the LAAW in the control group. The beta-NGF protein level was positively correlated with the density of sympathetic nerves in all groups. Probucol decreased the increase of CRP concentration and attenuated atrial oxidative stress caused by atrial tachypacing. In addition, probucol could effectively inhibit atrial beta-NGF upregulation, significantly attenuate atrial nerve sprouting and heterogeneous sympathetic hyperinnervation, and dramatically reduce the inducibility and duration of AF. CONCLUSIONS: The atrial over expression of beta-NGF possibly caused by increased oxidative stress and inflammation may be the main mechanism underlying atrial autonomic remodeling during AF. Probucol attenuates atrial autonomic remodeling possibly by its antioxidant and anti-inflammatory actions. PMID- 19187622 TI - Preliminary separation of the growth factors in platelet-rich plasma: effects on the proliferation of human marrow-derived mesenchymal stem cells. AB - BACKGROUND: Platelet-rich plasma (PRP) as a storage vehicle of growth factors has been successfully used in clinical applications, but in most cases the platelets were autologous. However, the large volume of blood withdrawn has detrimental effects on patients with anemia or poor general health. To overcome these limitations, this study was designed to separate the growth factors in homologous platelet-rich plasma. METHODS: The gel chromatography with Superdex-75 column was applied to separate PRP supernatants into 4 major fractions. Then the four fractions were vacuumed freeze-dried and re-dissolved in phosphate buffered saline. Proteins concentrations in PRP and in four fractions were detected by bicinchoninic acid protein assay; platelet derived growth factor-AB (PDGF-AB) and transforming growth factor beta1 (TGF-beta1) levels were determined by sandwich enzyme-linked immunosorbent assays. The effects of fractions on the proliferation of human marrow-derived mesenchymal stem cells (MSCs) were determined by 3-(4, 5- dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. RESULTS: PRP supernatants were separated into four major fractions by gel chromatography. The proteins recovery was 96.72%. Of the four fractions, fraction B contained the highest TGF-beta1 and PDGF-AB levels, and the highest proteins concentrations. Cell proliferation curves of MSC demonstrated that fraction B and C induced a remarkable increase of MTT values compared to the untreated culture (P < 0.05), and the effects of fraction B and C showed no significant difference compared to the PRP group (P > 0.05). Fraction A and D showed no significant difference to the negative control group (P > 0.05). CONCLUSIONS: The growth factors in PRP supernatants could be preliminarily separated into four fractions by gel chromatography, and the freeze-drying fractions retained the biological activity of growth factors. The growth factors were mostly presented in fraction B and C, and they promoted cell proliferation effectively. PMID- 19187623 TI - Clinical outcomes of serolimus-eluting stents versus bare metal stents in ST segment elevation myocardial infarction patients: a meta-analysis. AB - BACKGROUND: The benefits and safety of sirolimus-eluting stent (SES) have not been systematically quantified in different trials in ST-segment elevation myocardial infarction (STEMI) patients with primary or rescue percutaneous coronary intervention (PCI). A meta-analysis of randomised trials comparing SES and bare-metal stent (BMS) was performed. METHODS: A systematic literature search was conducted to identify all randomized clinical trials. The primary outcome was the rate of major adverse cardiac events (MACEs). The secondary outcomes included death, recurrent myocardial infarction, recurrent revascularization, and stent thrombosis. RESULTS: Totally, 1973 STEMI patients were enrolled in seven eligible randomized trials comparing SES with BMS. The pooled rate of major adverse cardiac events was significantly lower in the SES group than in the BMS group (9.7% vs 20.3%, OR 2.45, 95% CI 1.88-3.19, P < 0.00001). No significant difference in all causes of death was found between the SES and BMS groups, as well as in the pooled recurrent myocardial infarction rates. The pooled recurrent revascularization rate was significantly lower in the SES group than in the BMS group (5.1% vs 14.8%, OR 3.30, 95% CI 2.37-4.60, P < 0.00001). No significant difference was found between the pooled rates of stent thrombosis (1.2% in the SES group and 2.0% in the BMS group, OR 1.61, 95% CI 0.79-3.26, P = 0.19). CONCLUSIONS: SES is associated with a decreased risk of major adverse cardiac events compared with BMS by the greater reduction in repeat revascularization in STEMI patients. Larger trials with longer follow up are warranted to better define the role of SES in STEMI. PMID- 19187625 TI - Vaccination against hepatitis B: the Chinese experience. AB - OBJECTIVE: To review the implementation of mass vaccination of hepatitis B vaccine and its critical role in prevention of hepatitis B virus infection in China. DATA SOURCES: The data were mainly from PubMed, China Hospital Knowledge Database, and other popular Chinese journals published from 1980 to 2008. The search term was "hepatitis B vaccine". STUDY SELECTION: Original studies conducted in China and critical reviews authored by principal investigators in the field of hepatology in China were selected. RESULTS: Chinese investigators started to develop hepatitis B vaccine in late 1970s. The first home-made plasma derived vaccine became available in 1986, which has been completely replaced by the domestically produced recombinant (yeast or Chinese hamster ovary cell) vaccine since 2001. China health authority recommended vaccinating all infants in 1992. From then on, China has put tremendous efforts in implementation of mass vaccination. The overall coverage of hepatitis B vaccine in infants has increased steadily and reached more than 95.0% in urban and 83.0% - 97.0% in rural areas. The chronic HBV carrier rate in children < 10 years of age decreased from 10.0% before the mass vaccination to 1.0% - 2.0% in 2006, and that in general population decreased from 10.0% to 7.2%; overall, the nationwide mass hepatitis B vaccination has reduced more than 30 million of chronic HBV infections and HBV related severe sequlae. CONCLUSION: The Chinese successful experience in control of hepatitis B by mass vaccination offers an example for any unindustrialized country whoever is committed to control this disease. PMID- 19187624 TI - Human immunodeficiency virus and hepatitis C virus co-infection: epidemiology, natural history and the situation in China. PMID- 19187626 TI - Postmastectomy radiation in supraclavicular and internal mammary regions of patients with breast cancer of stage II/III. PMID- 19187627 TI - Angiotensin II vaccine promising for patients with chronic heart failure. PMID- 19187628 TI - Malignant transformation of aneurysmal bone cysts: a case report. PMID- 19187629 TI - Ultimate treatment for a patient with severe traumatic brain injury without intake for 192 hours after Wenchuan massive earthquake. PMID- 19187631 TI - Has the time come for another breakthrough in surgical myocardial revascularization? PMID- 19187632 TI - Current status of coronary artery bypass surgery. PMID- 19187633 TI - Bilateral bidirectional superior cavopulmonary shunt is more beneficial in medium and long term clinical outcomes than unilateral shunt. AB - BACKGROUND: The present study was aimed to compare the effects of bilateral and unilateral bidirectional superior cavopulmonary shunt (b-BDG and u-BDG) on pulmonary artery growth and clinical outcomes. METHODS: The 51 subjects enrolled in this study were divided into two groups: those receiving b-BDG (n = 21) and those receiving u-BDG (n = 30). Clinical records were reviewed retrospectively at a mean of 43.3 months after BDG procedures. Chi square and t-tests were performed to analyze the data. RESULTS: Left and right pulmonary artery diameters increased 27% - 37% in both groups. The pulmonary artery index increased 37.2% after b-BDG and 27.0% after u-BDG. b-BDG patients experienced a significant decrease in mean hemoglobin concentration and hematocrit value, and a correlated change in postoperative diameter of left pulmonary artery (LPA) and pulmonary artery index (y = 0.2719, x = -1.8278; R = 0.564, P = 0.008). The change ratio of hemoglobin and postoperative LPA were also correlated in b-BDG patients (y = -0.0522x + 0.3539; R = -0.479, P = 0.028). Only one b-BDG patient versus twelve u-BDG patients needed total cavopulmonary connections 31.8 months after BDG surgery (P = 0.0074). Moreover, only one (4.8%) b-BDG patient but eight u-BDG patients (26.7%) developed pulmonary arteriovenous malformations. CONCLUSIONS: b-BDG increases bilateral pulmonary blood flow and promotes growth of bilateral pulmonary arteries, with preferable physiological outcomes to u-BDG. Results may imply that subsequent Fontan repair may not always be needed. PMID- 19187634 TI - Using intramyocardial electrograms combined with other noninvasive methods for monitoring acute rejection following human heart transplantation. AB - BACKGROUND: Acute allograft rejection in heart transplantation remains as one of the major complications. Obligatory graft surveillance is still achieved with the invasive and expensive endomyocardial biopsy (EMB). Our study aimed to study the use of intramyocardial electrograms combined with other noninvasive methods for the monitoring of acute rejection after human heart transplantation. METHODS: Permanent pacemakers were implanted in 58 patients undergoing heart transplantations. Intramyocardial electrograms (IMEG) were recorded periodically and the results were compared with those from EMBs. The R wave amplitude of the IMEG was used as the index value, the average R wave amplitude at the third week following transplantation was considered as the baseline, and a reduction of > 20% compared with the baseline was regarded as a positive result. EMB was performed in cases of positive IMEG results and also at other times. Other noninvasive methods were used to help the diagnosis. Acute rejection (AR) was defined as International Society of Heart-Lung Transplantation grade IIIA or higher. RESULTS: We obtained 1231 IMEG records and 127 EMBs. Of the total 127 EMBs, 53 were positive, in which there were 42 IMEG positive results and 11 negative, while in the rest 74 negative EMBs, there were 9 IMEG positive results and 65 negative. The sensitivity of IMEG for the diagnosis of AR was 79.2%, and the specificity was 87.8%. The positive predictive value was 82.4% and the negative predictive value was 85.5%. Of the total of 1231 IMEG records, 51 were positive and 1180 were negative. Excluding 11 proved by EMB to be false negative, if the other 1169 were considered as no evidence of rejection, through the other noninvasive methods, AR diagnosed by this noninvasive monitoring strategy, the sensitivity was 79.2%, and the specificity was 99.2%. The positive predictive value was 82.4% and the negative predictive value was 99.1%. CONCLUSIONS: IMEG can be used as a noninvasive method for monitoring AR following heart transplantation. It is a continuous, safe and inexpensive method, and could reduce the need for EMB combined with other noninvasive methods, without reducing the detection of rejection. PMID- 19187635 TI - Clinical study of domestic ZQL-type retrievable vena cava filter. AB - BACKGROUND: Pulmonary thromboembolism (PTE) is a serious disease often leading to disability and death. Percutaneous placement of an inferior vena cava (IVC) filter is an effective method to prevent fatal PTE caused by lower extremity deep venous thrombosis (LEDVT). We developed the ZQL-type retrievable vena cava filter. The aim of this study was to evaluate the safety and effectiveness of the ZQL-type filter for prevention of fatal PTE. METHODS: A total of 144 patients with indications for placement of an IVC filter received insertion of filters via a femoral (n = 37) or jugular (n = 107) vein approach. Abdominal X-ray and color Doppler ultrasonography of IVC were regularly performed to visualize the position and condition of the IVC filter following filter placement. If thrombi in the lower extremity deep veins were removed within 2 weeks, the filter retrieval procedure was performed after an abdominal radiograph, an inferior vena cavogram and a pulmonary angiography. Otherwise, the filter should be kept permanently in place with regular follow-up inspections. RESULTS: One hundred and forty-four filters were implanted at the target sites with a success rate of 98.61% for one time placement and 100% for two-time placement. Of the 137 patients followed up (follow-up rate, 95.14%), 43 patients had filters retrieved during a period of between 7 and 14 days (median, 12 days) following filter placement. The remaining 94 patients were followed for 7 days to 39 months (median follow-up period, 17 months). No filter migration or tilt, filter fracture or IVC perforation was observed. No obstruction of IVC occurred. No symptomatic PTE developed during the follow-up period. CONCLUSION: The ZQL-type retrievable vena cava filter is a safe and effective device to prevent PTE. This filter possesses a distinctive stent shape, stable design, high capture efficacy, and is easy to insert and retrieve and thus is suitable for clinical application. PMID- 19187636 TI - Assessment of the anterior spinal artery and the artery of Adamkiewicz using multi-detector CT angiography. AB - BACKGROUND: Damage to the spinal cord after the treatment of the descending thoracic and thoracoabdominal aortic aneurysms is an uncommon but devastating complication. The artery of Adamkiewicz (AKA) is the principal arterial supply of the anterior spinal artery (ASA) in the lower thoracic and lumbar level. The purpose of this study was to evaluate the visualization of the anterior spinal artery and the artery of Adamkiewicz, the affecting factors for the detection rate using multi-detector row CT (MDCT). METHODS: Ninety-nine consecutive patients (31 women and 68 men; age range, 25 - 90 years; average age 61.3 years), with suspicion for thoracic aortic lesions necessitating surgical intervention (31 aortic aneurysm, 45 dissection, 5 intramural hematoma, and 18 normal), underwent CT angiography from the aortic arch to the aortic bifurcation. Transverse sections, multiplanar reformations and thin maximum intensity projections were used to assess the ASA and AKA. The level of the ASA and AKA origins and CT acquisition parameters were recorded. The contrast-to-noise ratio of the image, an index of the mass of the T11 body (vertebral mass index), the subcutaneous fat thickness, and the CT value within the aortic arch and at the T11 level were measured. The detection of the ASA and AKA were evaluated relative to the acquisition parameters, scan characteristics, and aortic lesion type. Differences were assessed with the Wilcoxon rank-sum and t tests. RESULTS: The ASA was visualized in 51 patients (52%) and the AKA in 18 patients (18%). The ASA was identified in 36/67 patients (54%) with 1.25 mm thickness and in 15/32 patients (47%) with 2.5 - 3.0 mm thickness. This difference did not achieve significance (P = 0.13). The detection rate of the ASA and the AKA was influenced by the vertebral mass index and the contrast-to-noise ratio (P < 0.05). The amount of subcutaneous fat affected the detection rate of the ASA (P < 0.05) but not the AKA. In CT scans of ASA detection, the mean CT values in the aorta at the arch and at T11 were 360 and 358 HU, respectively, whereas in CT scans without ASA detection, the CT values in the aorta at the arch and at T11 were lower (P < 0.05), 297 and 317 HU, respectively. CONCLUSIONS: The ASA and AKA were less frequently detected in our cohort than previous reports. The visualization of the ASA and AKA was significantly affected by aortic enhancement, the "vertebral mass index", and the contrast-to-noise ratio. PMID- 19187637 TI - Effect of myocardial protection during beating heart surgery with right sub axiliary approach. AB - BACKGROUND: Cardiac troponin-I (cTnI) is one of the three regulatory subunits of the cardiac troponin which has the high sensibility and specificity of responding to myocardial injury. Studies have demonstrated that cTnI is released into the blood stream within hours following acute myocardial reperfusion injury. The clinical utility of cTnI for the assessment of myocardial damage is that it is more specific than creatine kinase MB (CKMB). This study investigated cTnI as a sensitive marker of myocardial reperfusion injury and its clinical value on beating heart surgery with right sub-axiliary incision. METHODS: From December 2002 through December 2004, 100 patients with atrial septal defect (ASD), ventricular septal defect (VSD), atrial septal defect and ventricular septal defect (ASD + VSD), and tetralogy of Fallot were randomly divided into two groups: the treatment group (n = 50) was operated on with a beating heart under extracorporeal circulation (ECC), and the control group (n = 50) on an conventional arresting heart under ECC. The two groups both used a right sub axillary incision. Blood samples from a central venous catheter (CVC) were collected before, at the end of aortic clamping, immediately after discontinue cardiopulmonary bypass (CPB), 3, 6, 24, and 48 hours after operation. The Abbott Axsym system with hol-automation fluorescent immunity analyzer was used for the quantitative determination of cTnI. cTnI was detected to investigate the effect of myocardial ischemia reperfusion injury and the clinical value of beating heart surgery with right sub-axillary incision. RESULTS: There were no significant differences between the two groups before operation. At the end of aortic clamping and thereafter, cTnI significantly increased in both groups, and reached the peak point at 6 hours after operation. At all the tested points, cTnI was significantly higher in the control group than the beating heart group (P < 0.05), especially at 6 hours post operation (P < 0.01). The operating time and ECC duration were shortened and the dosage of dopamine was decreased, when compared with the control group. CONCLUSIONS: There was less cTnI measured in the beating heart group than in the control group after CPB, demonstrating that beating heart surgery may significantly reduce myocardial reperfusion injury. PMID- 19187638 TI - Decreased expression of neurotrophic tyrosine receptor kinase 3 is associated with the outflow tract defect of human tetralogy of Fallot. AB - BACKGROUND: The molecular mechanism of human tetralogy of Fallot (TOF) is incompletely defined. Animal models have suggested that neurotrophic tyrosine receptor kinase 3 (NTRK3) might be associated with the outflow tract defect, similar to that seen in human TOF, however, the expression pattern of NTRK3 in human TOF heart tissues has never been investigated. METHODS: Quantitative real time PCR (qRT-PCR) and immunohistochemistry were applied to detect NTRK3 mRNA and protein levels in right ventricular outflow tract tissue samples of TOF patients, ventricular septal defect (VSD) patients and normal control infants (n = 10 in each group). RESULTS: qRT-PCR analysis indicated that NTRK3 mRNA levels were significantly decreased in the TOF group compared to the VSD group (0.024 +/- 0.003 vs 0.085 +/- 0.004, P = 0.022) and the normal control group (0.024 +/- 0.003 vs 0.091 +/- 0.002, P = 0.006). Quantitative immunohistochemical analysis showed that NTRK3 protein was mainly localized in the myocardium cytoplasm in all 3 groups. The immunoreactivity of NTRK3 protein was again significantly lower in the TOF group compared to the VSD group (1.42 +/- 0.62 vs 14.12 +/- 1.83, P = 0.023) and the control group (1.42 +/- 0.62 vs 16.25 +/- 2.31, P = 0.008). The expression of NTRK3 in the VSD group and in the control group showed no significant differences at both mRNA and protein levels. CONCLUSIONS: Insufficient expression of NTRK3 is associated with the outflow tract defect of human tetralogy of Fallot and may contribute to the progression of this defect. PMID- 19187639 TI - Coronary stenting does not improve the long-term cardiovascular outcome of patients with mild to moderate renal insufficiency. AB - BACKGROUND: Several studies have shown that coronary stenting reduces the frequency of clinical and angiographic restenosis in patients with mild to moderate renal insufficiency. However, less is known about the long-term benefits of stent use in this population. This study was aimed to determine the impact of coronary stenting on extended (5 years) long-term outcomes of patients with chronic renal insufficiency. METHODS: The study included 602 consecutive patients who underwent successful percutaneous coronary intervention with stenting. Renal insufficiency was defined as an estimated glomerular filtration rate < 60 ml x min(-1) x 1.73 m(-2). The major adverse cardiac events were compared for patients with (n = 160) and without (n = 442) renal insufficiency. RESULTS: After the third year of follow-up, nonfatal myocardial infarction and revascularization rates were significantly increased in patients with renal insufficiency compared with those without renal dysfunction (16.9% vs 7.7%, P = 0.001; 29.4% vs 15.8%, P < 0.001). In patients who had recurrent cardiovascular events, a significantly higher rate of de novo stenosis revascularization was found in patients with renal insufficiency than without renal insufficiency (57.7% vs 22.7%, P < 0.001), while there was no significant difference in target lesion revascularization between the groups (51.9% vs 43.6%, P = 0.323). Multivariate analysis demonstrated an independent impact of the presence of renal insufficiency on the major adverse cardiac events (hazard ratio: 1.488, 95% confidence interval: 1.051 - 2.106, P = 0.025) and de novo stenosis (hazard ratio: 5.505, 95% confidence interval: 2.151 - 14.090, P < 0.001). CONCLUSIONS: The late major adverse cardiac events, after successful coronary stenting, is increased in patients with an estimated glomerular filtration rate < 60 ml x min(-1) x 1.73 m(-2). This might be associated with increased risk of de novo stenosis in this population. PMID- 19187640 TI - The utilization status of aspirin for the secondary prevention of ischemic stroke. AB - BACKGROUND: The present study was aimed to investigate the usage of aspirin for the secondary prevention of ischemic stroke, evaluate the correlated factors, and analyze the reasons for not taking and irregularly taking aspirin. METHODS: The patients in this group were all stroke survivors who have formerly been diagnosed with a cerebral infarction or transient ischemic attack (TIA) in our hospital. We investigated their use of aspirin over a three-year period following their hospitalization. According to the patients' aspirin usage, they were divided into treatment and non-treatment groups. In addition, the reasons for not taking or irregularly taking aspirin were analyzed in the two groups. RESULTS: A total of 1240 patients were studied, including 367 (29.60%) in the treatment group and 873 (70.40%) cases in the non-treatment group. In addition, 201 (16.20%) cases in the treatment group had been regularly taking aspirin (50 - 325 mg of aspirin daily) for 1 to 3 years or longer. The results demonstrated that the main reasons for not taking aspirin in this study were related to patients' concerns regarding the side effects of taking aspirin (46.45%), as well as the doctors' inadequacy in informing their patients to take aspirin (38.71%). The major reasons for patients to irregularly take aspirin were that the doctors did not notify the length of aspirin usage to their patients (41.57%), and that doctors did not prescribe aspirin upon the patients' follow-up visit (26.51%). CONCLUSION: The most effective way to increase patient's compliance for aspirin consumption is to promote the guidelines for stroke treatment and to relay these advances in stroke therapy to the patient. PMID- 19187641 TI - Fluid therapy for severe acute pancreatitis in acute response stage. AB - BACKGROUND: Fluid therapy for severe acute pancreatitis (SAP) should not only resolve deficiency of blood volume, but also prevent fluid sequestration in acute response stage. Up to date, there has not a strategy for fluid therapy dedicated to SAP. So, this study was aimed to investigate the effects of fluid therapy treatment on prognosis of SAP. METHODS: Seventy-six patients were admitted prospectively according to the criteria within 72 hours of SAP onset. They were randomly assigned to a rapid fluid expansion group (Group I, n = 36) and a controlled fluid expansion group (Group II, n = 40). Hemodynamic disorders were either quickly (fluid infusion rate was 10 - 15 ml x kg(-1) x h(-1), Group I) or gradually improved (fluid infusion rate was 5 - 10 ml x kg(-1) x h(-1), Group II) through controlling the rate of fluid infusion. Parameters of fluid expansion, blood lactate concentration were obtained when meeting the criteria for fluid expansion. And APACHE II scores were obtained serially for 72 hours. Rate of mechanical ventilation, incidence of abdominal compartment syndrome (ACS), sepsis, and survival rate were obtained. RESULTS: The two groups had statistically different (P < 0.05) time intervals to meet fluid expansion criteria (Group I, 13.5 +/- 6.6 hours; Group II, (24.0 +/- 5.4) hours). Blood lactate concentrations were both remarkably lower as compared to the level upon admission (P < 0.05) and reached the normal level in both groups upon treatment. It was only at day 1 that hematocrit was significantly lower in Group I (35.6% +/ 6.8%) than in Group II (38.5% +/- 5.4%) (P < 0.01). Amount of crystalloid and colloid in group I ((4028 +/- 1980) ml and (1336 +/- 816) ml) on admission day was more than those of group II ((2472 +/- 1871) ml and (970 +/- 633) ml). No significant difference was found in the total amount of fluids within four days of admission between the two groups (P > 0.05). Total amount of fluid sequestration within 4 days was higher in Group I ((5378 +/- 2751) ml) than in Group II ((4215 +/- 1998) ml, P < 0.05). APACHE II scores were higher in Group I on days 1, 2, and 3 (P < 0.05). Rate of mechanical ventilation was higher in group I (94.4%) than in group II (65%, P < 0.05). The incidences of abdominal compartment syndrome (ACS) and sepsis were significantly lower in Group II (P < 0.05). Survival rate was remarkably lower in Group I (69.4%) than in Group II (90%, P < 0.05). CONCLUSIONS: Controlled fluid resuscitation offers better prognosis in patients with severe volume deficit within 72 hours of SAP onset. PMID- 19187642 TI - Intraoperative monitoring for safety of total hip arthroplasty using third generation cementing technique. AB - BACKGROUND: Controversies on the safety of the cement application between cemented and uncemented total hip arthroplasty (THA) have been existing for decades. The purpose of this study was to observe the changes in mean arterial pressure (MAP), heart rate (HR) and oxygen pressure (PaO(2)) during cemented THA, and to evaluate the intraoperative safety of using the third-generation cementing technique and investigate whether the intraoperative risk is higher in acute femoral neck fracture patients than non-traumatic patients. METHODS: Forty-two patients who underwent cemented THA between November 2005 and September 2007 were prospectively included in this study. The third-generation cementing technique as vacuum mixing and pulsatile lavage was used strictly. The MAP and HR were monitored and documented during each operation. Blood gas analysis was performed at exposure, cup implantation, stem implantation and wound closure. MAP, HR and PaO(2) were compared between pre- and post-cement application. Comparisons of MAP, HR and PaO(2) between patients with acute femoral neck fracture and non traumatic patients were performed as well. RESULTS: No intraoperative cardiopulmonary complication occurred in these cases. No obvious changes were observed in MAP, HR and PaO(2) after cement application. There was no significant difference in MAP, HR and PaO(2) between acute femoral fracture patients (18 patients) and non-traumatic patients (24 patients). CONCLUSIONS: The results of this study suggested that the invasive blood pressure monitoring and blood gas analysis are essential for patients undergoing cemented THA, especially for patients with femoral neck fracture. The third-generation cementing technique is safe to use in THA. PMID- 19187643 TI - A new study on diffusion tensor imaging of the whole visual pathway fiber bundle and clinical application. AB - BACKGROUND: With conventional imaging methods only the morphous of the visual nerve fiber bundles can be demonstrated, while the earlier period functional changes can not be demonstrated. We hypothesized that diffusion tensor imaging (DTI) would demonstrated the whole optic never fiber bundle and visual pathway and the earlier period functional changes. The purpose of the present study was to evaluate the application of DTI technique in the demonstration of the whole optic never fiber bundle and visual pathway, and the influence of orbital tumors on them. METHODS: GE 1.5 T signa HD MR System, and the software package DTV2 were adopted. The total 45 subjects were enrolled, including 15 volunteers and 30 patients. All patients had ocular proptosis from minor to major. Seven patients had visual acuity decrescence. RESULTS: The nerve fiber bundles, e.g. optic chiasma, optic tract and optic radiation in posterior visual pathway were well demonstrated in all cases. Wherein, the intact whole visual pathway fiber bundles were clearly revealed in 10 volunteers and 17 patients, and optic nerve was not wholly revealed in the rest of the subjects. Shift of optic nerve caused by compression and partial deformation were seen in 7 patients with orbital tumor. In 6 of 7 patients, DTI displayed significant abscise and deformation of visual nerve. Chi-square test indicated significant correlation between visual acuity decrescence and DTI visual nerve non-display. CONCLUSIONS: Visual nerve fiber bundles and the whole visual pathway were visualized in most of patients with DTI. It might be an effective method of providing imaging evidence for visual nerve fiber earlier period functional changes, and laid a foundation for the study in other cranial nerves. PMID- 19187644 TI - Chemokine stromal cell-derived factor 1/CXCL12 increases homing of mesenchymal stem cells to injured myocardium and neovascularization following myocardial infarction. AB - BACKGROUND: Heart failure due to ischemic heart disease is still a major health problem. Myocardium regeneration emerges as a novel therapeutic method for treating myocardial infarction (MI). However, it is affected by many factors. The present study was aimed to investigate the effect of chemokine stromal cell derived factor 1 (SDF-1)/CXCL12 on mesenchymal stem cells (MSCs) homing to injured myocardium in a rat myocardial infarction model. METHODS: A rat myocardial infarction model was established by permanent left anterior descending branch ligation. Mesenchymal stem cells from donor rats were cultured in IMDM and labeled with bromodeoxyuridine. The rats were divided into two groups. SDF-1 expression was measured by in situ hybridization and immunohistochemistry in the sham operated or infarcted hearts at 1, 2, 4, 7, 14 and 28 days post operation in the SDF-1 detection group. The rats in the intervention groups were injected with SDF-1, anti-SDF-1 antibody or saline 4 days after myocardial infarction. Then, a total of 5 x 10(6) cells in 2.5 ml of phosphate-buffered saline were injected through the tail vein. The number of the labeled MSCs in the infarcted hearts was counted on the 3rd day post injection. Cardiac function and blood vessel density were assessed on the 28th day post injection. RESULTS: Self-generating SDF-1 expression was increased at the first day post MI, peaked at the 7th day and decreased thereafter while it remained unchanged in sham operated hearts. The MSCs enrichment in the host hearts were more abundant in the MI groups than in the non-MI group (P = 0.000); the MSCs enrichment in the host hearts were more abundant in the SDF-1 injected group than in the anti-SDF-1 antibody and saline injected groups (P = 0.000). Cardiac function was improved more in the SDF-1 injected group than in the anti-SDF-1 antibody and saline injected groups (P = 0.000). Neovascularization in the SDF-1 injected group increased significantly compared to the other groups (P = 0.000). CONCLUSION: Myocardial SDF-1 expression was increased only in the early phase post MI. SDF-1 may enhance MSCs homing to the injured heart and improve cardiac function by promoting neovascularization. PMID- 19187645 TI - Celecoxib plays a multiple role to peripheral blood lymphocytes and allografts in acute rejection in rats after cardiac transplantation. AB - BACKGROUND: Celecoxib, a selective cyclooxygenase-2 (COX-2) inhibitor, is a non steroidal anti-inflammatory drug used as an adjuvant to sensitize cancer cells to apoptosis. However, in rats suffering from acute rejection, celecoxib reduced apoptosis of myocardial cells. We hypothesize that celecoxib reduces myocardial apoptosis either by inducing apoptosis in peripheral blood lymphocytes (PBLs) or by altering the percentage of CD4(+) and CD8(+) lymphocytes. METHODS: After cardiac transplantation, rats were administered intragastrically with celecoxib (50 mg/kg per day) for 3, 5 or 7 days, at which time the graft was excised and evaluated for organ rejection. In addition, PBLs were isolated from the blood to determine PBLs apoptosis, and the percentage of CD4(+) and CD8(+) lymphocytes. RESULTS: Celecoxib induced PBLs apoptosis in 3 days, but protected the cells from apoptosis at 5 and 7 days. Also, the percentage of CD4(+) lymphocytes decreased only at 3 days, but a reduction in the percentage of CD8(+) lymphocytes was not seen until 7 days after the transplant surgery. Celecoxib only decreased acute rejection at 5 days, with no discernible difference in rejection after 3 and 7 days. CONCLUSIONS: The results suggested that celecoxib displayed a multiple physiological function in a time-dependent manner. PMID- 19187646 TI - Strain rate imaging in assessing the size of acute ischemic myocardium in dogs. AB - BACKGROUND: Since the size of ischemic myocardium is closely related with both global and regional function of the myocardium, it is of great significance to measure the size of ischemic myocardium with non-invasive methods. METHODS: Eleven mongrel dogs were subjected to occlusion of the left anterior descending coronary artery for acute ischemia. Strain rate imaging had M-mode of strain-rate (CAMM) curve pointed from the basal segment of the anterior wall to the basal segment of the inferior wall to detect the border of ischemia size. The strain rate (SR) defined the cut-off value of ischemic myocardium in a two-chamber apical view, and marked by the anterior and inferior wall on two-dimensional images respectively. Along the endocardium and epicardium, the ischemic size was curved on two-dimensional images by the trackball method and then compared with the pathologically ischemic size. And then longitudinal strain rates were compared in the cut-off value, adjacent non-ischemic and ischemic segments at which the cut-off point was defined by changing the curve M-mode of strain rate after ischemia. RESULTS: Linear correlation existed between pathology and strain rate ischemic size (r = 0.884, P < 0.001). The SR parameters were lower in ischemia and cut-off point than in non-ischemic segments. The peak SRs of systole (S(SR)), early diastole (E(SR)), late diastole (A(SR)), strain during ejection time (epsilon(et)), and the maximum length change during the entire heart cycle (epsilon(max)) in ischemic segments lowered (P < 0.05). Time to onset of regional relaxation (T(R)) was prolonged (P = 0.012). CONCLUSION: SR imaging can accurately assess the size of ischemic myocardium. PMID- 19187647 TI - Could mycophenolate mofetil combined with benazapril delay tubulointerstitial fibrosis in 5/6 nephrectomized rats? AB - BACKGROUND: It was reported that combination of mycophenolate mofetil (MMF) and enalapril could reduce proteinuria, improve renal function, and down-regulate diabetes-induced macrophage recruitment and expression of monocyte chemotactic protein 1 (MCP-1) and transforming growth factor beta (TGF-beta) in diabetic renal tissue. But there are no compelling data available for the combination of MMF and angiotensin converting enzyme inhibitor (ACEI) for suppressing tubulointerstitial fibrosis in chronic kidney diseases. The present study was to disclose the effect of MMF combined with benazapril on delaying tubulointerstitial fibrosis and its possible mechanisms in 5/6 nephrectomized rats. METHODS: Fifty male SD rats underwent 5/6 nephrectomy (5/6 NX) were randomized into the following groups: NX (5/6 nephrectomized rats, distilled water, n = 10), MMF (MMF 20 mg x kg(-1) x d(-1), p.o., n = 10), Ben (benazepril 10 mg x kg(-1) x d(-1), p.o., n = 10), MMF/Ben (MMF 20 mg x kg(-1) x d(-1), p.o., and benazapril 10 mg x kg(-1) x d(-1), p.o., n = 10). They were monitored for proteinuria and systolic blood pressure every two weeks. After 8 weeks of treatment, serum creatinine and blood urea nitrogen were assayed and pathological damage to the kidney were evaluated. Renal expression and serum levels of platelet-derived growth factor-BB (PDGF-BB), matrix metalloproteinase-9 (MMP-9), and tissue inhibitor of metaloproteinase-1 (TIMP-1) were detected by immunohistochemistry and ELISA methods. RESULTS: After 8 weeks of treatment, 24 hour proteinuria, serum creatinine and blood urea nitrogen were significantly lower in treated groups compared with the untreated rats. MMF and benazepril combination therapy had a greater effect than either drug alone. MMF alone had no effect on systolic blood pressure, but benazapril and MMF/benazapril could significantly reduce blood pressure. Rats that underwent 5/6 nephrectomy had greater tubulointerstitial inflammatory cell infiltration and collagen accumulation than sham-operated rats; all treatments, especially MMF/benazepril, ameliorated these effects. Tubules in 5/6 nephrectomized rats expressed higher levels of PDGF-BB and TIMP-1 and lower MMP-9 compared with sham-operated rats. MMF and benazepril similarly reversed these phenomenons and combination therapy almost completely restored the expression of these cytokines in renal tissue and their plasma concentration. CONCLUSIONS: MMF, especially combined with benazepril, can reduce proteinuria, improve renal function, ameliorate tubulointerstitial fibrosis in 5/6 nephrectomized rats. These effects might be, in part, associated with down-regulation of PDGF-BB and TIMP-1, and MMP-9 up regulation in renal tissues. PMID- 19187648 TI - The regulating role of mutant IkappaBalpha in expression of TIMP-2 and MMP-9 in human glioblastoma multiform. AB - BACKGROUND: Our previous studies demonstrated that mutant IkappaBalpha (IkappaBalphaM) inhibited the occurrence, growth and angiogenesis of human glioblastoma multiform (GBM). However, the specific mechanism by which IkappaBalphaM regulates protein-degrading enzymes secreted from GBM to inhibit invasion and metastasis has remained unclear. The aim of the present study was to investigate the regulatory role and significance of IkappaBalphaM genes in the expression of tissue inhibitor of metalloproteinase (TIMP)-2 and matrix metalloproteinase (MMP)-9 in human GBM. METHODS: We established the following four GBM cell lines stably expressing IkappaBalphaM by plasmid construction, gene transfection and screening for IkappaBalphaM protein expression: mutant IkappaBalpha-transfected cells (G36Delta-M), wild-type IkappaBalpha-transfected cells (G36Delta-W), empty plasmid transfected cells (G36Delta-P) and untransfected cells (G36Delta). The TIMP-2 and MMP-9 expression was detected by RT-PCR and Western blotting. Tumor cells were then implanted subcutaneously into nude mice to establish an animal model of ectopic tumor growth, and TIMP-2 and MMP-9 expression was determined by immunohistochemical methods. RESULTS: The results showed that there was a significant increase in TIMP-2 expression and a significant decrease in MMP-9 expression in the G36Delta-M group at both the RNA and protein levels compared with the G36Delta-W group, G36Delta-P group and G36Delta group. Similar results were observed in the immunohistochemical staining analysis of tumor tissues. In the G36Delta-M group, TIMP-2 expression was significantly higher while MMP-9 expression was significantly lower than in the other three groups. CONCLUSIONS: Our findings indicate that IkappaBalphaM inhibits the activation of NF-kappaB. It significantly up-regulates TIMP-2 expression in human malignant glioma cells and down-regulates the expression of MMP-9. Thus, IkappaBalphaM maintains the integrity of the extracellular matrix and further inhibits the growth and metastasis of tumor tissues. PMID- 19187649 TI - Effects of prior freezing or drying on the swelling behaviour of the bovine cornea. AB - BACKGROUND: Frozen or dried corneal grafts are commonly used for stromal transplantation such as lamellar keratoplasty (full or partial thickness), keratophakia, epikeratophakia. Structural properties are important for the final optical results of these surgeries but the effects of freezing/thawing and drying/rehydration on the properties of the stroma are known little compared with the corneal endothelium, mainly because of lack of non-invasive technique to evaluate the stromal structure. This study aimed to investigate the swelling and structural properties of the bovine corneal stroma following freezing or drying by X-ray diffraction which was a non-invasive technique and could give ultra structural information in hydrated tissues. METHODS: Bovine corneas were either frozen at -40 degrees C or dried to constant weight in a dessicator over silica gel. Swelling was carried out by placing the corneas into dialysis tubing and equilibrating them against various concentrations of polyethylene glycol (PEG) to obtain a range of tissue hydrations. This method minimises the loss of soluble tissue components during the swelling process. Synchrotron X-ray diffraction was used to measure the average intermolecular spacing, the interfibrillar spacing and the fibril diameter as a function of hydration. Changes in light scattering were detected using a microdensitometer. RESULTS: Freezing and thawing of the cornea caused an increase in light scattering by 63.9% at tissue hydration (H) = 3.4, and by 50.0% at H = 4.9. Repeated freezing and thawing causes further increased by 38.9% at the second time and another 36.0% at the third time (P < 0.05). There was a tendency for both the frozen and the dried corneas to lose some swelling ability, achieving hydrations respectively of 10% and 18% below those of fresh corneas at 0 PEG. There were no changes in the fibril diameters, interfibrillar or intermolecular spacings as measured by X-ray diffraction in the equilibrated fresh, pre-frozen and pre-dried corneas. CONCLUSIONS: The increase in light scattering and the loss of swelling ability after freezing and thawing probably results from structural changes following the close association of the collagen molecules and fibrils whilst the tissue is in the dry or frozen state. Some unknown changes in the extracellular matrix between the collagen fibrils may also play a role in the light scattering. The equilibration technique may improve the quality of rehydrated corneal graft or lenticules used for corneal surgeries. PMID- 19187650 TI - Classification and management of the tandem ossification of the posterior longitudinal ligament and flaval ligament. AB - OBJECTIVE: To provide appropriate guidelines for treatment of tandem ossification of the posterior longitudinal ligament (OPLL) and flaval ligament (OFL). Data sources Published articles about OPLL and OFL were selected using Medline and Embase electronic databases. Study selection An English literature search from January 1980 to December 2006 was conducted. Because many reported cases were incorporated in OFL studies, the key words for search were OFL or OFL and OPLL. The first step revealed 93 studies of which 13 reports of tandem OPLL and OFL (tandem ossification) were selected. RESULTS: All studies were case series or case report and advocated that the primary therapy for tandem ossification should be operative. The clinical outcomes of surgery were evaluated in most reports, predominantly using the JOA scores. Gender is the only factor which has prognostic value. A higher proportion of women was found in the failure group. A two-stage classification of tandem ossification was developed to relate diagnosis to outcome. CONCLUSIONS: All patients with suspected ossification of the spinal ligaments should undergo routine MRI screening of the whole spine. The correlation of the classification with surgical treatments needs further studies to validate its usefulness. PMID- 19187651 TI - Human umbilical cord mesenchymal stem cells and the treatment of spinal cord injury. AB - OBJECTIVE: To review the recent studies about human umbilical cord mesenchymal stem cells (hUCMSCs) and advances in the treatment of spinal cord injury. Data sources Published articles (1983 - 2007) about hUCMSCs and spinal cord injury were selected using Medline. Study selection Articles selected were relevant to development of mesenchymal stem cells (MSCs) for transplantation in spinal cord injury therapy. Of 258 originally identified articles 51 were selected that specifically addressed the stated purpose. RESULTS: Recent work has revealed that hUCMSCs share most of the characteristics with MSCs derived from bone marrow and are more appropriate to transplantation for cell based therapies. CONCLUSIONS: Human umbilical cord could be regarded as a source of MSCs for experimental and clinical needs. In addition, as a peculiar source of stem cells, hUCMSCs may play an important role in the treatment of spinal cord injury. PMID- 19187652 TI - Diagnosis and surgical treatment of post-infarction left ventricular pseudoaneurysm. PMID- 19187653 TI - A giant cyst-like mass: an unusual morphous of left atrial myxoma. PMID- 19187654 TI - Isolated superior mesenteric artery dissection: case for conservative treatment and endovascular repair. PMID- 19187655 TI - [To recognize monovision correctly and matching different types of intraocular lenses in two eyes]. AB - The production of different kinds of novel intraocular lens (IOLs) with various functions plays an active role in improving cataract surgery by providing more choices to meet different clinical demands. However, each type of IOL has its own advantages and disadvantages. Therefore, it is important to implant two different kinds of IOLs into two eyes of a patient, which can optimize the advantages of these two IOLs and can obtain a better mixing vision. But there are many misunderstandings about how to comprehend the principle of monovision and matching two different types of IOLs. When we consider monovision and matching two different types of IOLs, there are many challenges for us, e.g. to explain to the patients about the difference between expected and practical results, and the psychological adaptation requires a long period; and we should to avoid selection the IOLs blindly and to provide the best results and benefits for the patients. PMID- 19187656 TI - [Matching and selection of two different types of pseudo-accommodative intraocular lens in two eyes after cataract phacoemulsification]. AB - Refractive cataract surgery not only provides patients with a clear and comfortable far vision, but also improves near and intermediate vision to meet the patient's demands. The present study analyzed the results of separate implantations of two different types of intraocular lenses (IOL) into two eyes of cataract patients (using one type of IOL in one eye and another type of IOL in the contralateral eye). By knowing the patients different life-styles and occupations and familiarity with the advantages of various pseudo-accommodative IOL can allow the surgeon to select the proper types of pseudo-accommodative IOL in two different eyes for obtaining the best visual function. PMID- 19187657 TI - [Selection of accurate IOL formula in patients with cataract and high hyperopia]. AB - OBJECTIVE: To audit intraocular lens (IOL) power predictions for cataract surgery in high hyperopia by comparing the accuracy of different formulae of Haigis, SRKII, Hoffer Q, Hollady, and SRK/T. METHODS: A comparative study was used in 31 eyes with cataract and high hyperopia underwent phacoemulsification and posterior chamber IOL implantation. Eyes were examined with A-scan and IOL Master before the operation. Postoperative refraction was examined 3 months after surgery. We compared the accuracy of prediction between these different formulae using IOL Master, and then compared the errors using the two different methods for axial length measurement. RESULTS: (1) Using IOL Master, the Haigis formula showed the smallest mean prediction errors (0.37 +/- 0.14), followed by the Hoffer Q, Holladay, SRK/T, and SRK II formulae (-0.70 +/- 0.12, -0.97 +/- 0.15, -1.25 +/- 0.14, -1.46 +/- 0.13). The Haigis formula generated a slightly myopic result, but the other formulae generated hyperopic result at different degrees. (2) The A scan led to a prediction error that was toward positive values (equivalent to underestimating axial length), whereas the IOL Master error was towards the negative. Hoffer Q formula appeared to be more accurate when measuring axial lengths with A-Scan, whereas Haigis formula was more accurate when combined with IOL Master (0.37 +/- 0.14). CONCLUSIONS: For selection of IOL formula in cataract patients with high hyperopia, the Haigis would be the most accurate in IOL Master analysis, but the Hoffer Q was better when using A-scan. PMID- 19187658 TI - [Changes in high order aberrations of anterior and posterior surfaces of cornea before and after phacoemulsification]. AB - OBJECTIVE: To study the effects of phacoemulsification surgery on the optical high order aberrations (HOA) of the cornea measured with Pentacam. METHODS: It was a prospective case series study. HOAs of the anterior and posterior surface of cornea were measured before and after phacoemulsification surgery on 51 eyes of 45 patients. Monofocal foldable IOLs were implanted after phacoemulsification through a clear-cornea, 3.2 mm incision without suture. Patients were observed 1 day before surgery, and at 7 days, 1 month and 3 months after surgery. HOAs included the total high order aberration (tHOA), spherical aberration (SA), coma, trefoil, 3rd, 4th, 5th, 6th order aberrations. Root-mean-square (RMS) values of HOAs were obtained with 3 mm and 6 mm aperture using corneal topography of Pentacam pre- and postoperatively, and were divided into 3 mm and 6 mm aperture groups. Aberrations were compared in each eye using repeated measures and the optical changes induced by surgery were investigated. RESULTS: The total HOA of the anterior surface increased slightly from a mean of (0.289 +/- 0.171) microm preoperatively to (0.312 +/- 0.158) microm postoperatively with 3 mm aperture; and (0.980 +/- 0.215) microm preoperatively to (1.054 +/- 0.308) microm postoperatively with 6 mm aperture, respectively. The increase was not statistically significant (F = 1.55, P = 0.22), and the changing trend of the total HOA was similar between the two groups of 3 mm and 6 mm aperture (F = 0.95, P = 0.42). The changes of other HOAs were similar to the total HOA pre- and postoperative. The total HOA of the posterior surface increased significantly from (0.548 +/- 0.220) microm preoperatively to (0.661 +/- 0.275) microm postoperatively with 3 mm aperture, and (1.780 +/- 0.453) microm preoperatively to (1.945 +/- 0.532) microm postoperatively with 6 mm aperture. The increase was statistically significant (F = 15.83, P = 0.00). The changing trend of the total HOA showed statistically difference between these two groups (F = 12.46, P = 0.00). The changes of other HOAs were similar to the total HOA pre- and postoperatively. CONCLUSIONS: Phacoemulsification cataract surgery does not systematically degrade the optical quality of the anterior corneal surface. However, it introduces significant changes in most HOAs of the posterior surface of cornea, so the surgery may degrade the optical quality of the posterior corneal surface. PMID- 19187659 TI - [Pathological study on effects of preservative-free 1% lidocaine on lens epithelial cells of patients with age-related cataract]. AB - OBJECTIVE: To assess whether preservative-free 1% lidocaine is capable of destroying the LECs in age-related cataract (ARC) in order to provide scientific basis for pursuing safe and effective drugs to eliminate LECs in cataract surgery. METHODS: Lens anterior capsule (LAC) specimens were collected from 75 patients (82 eyes) with age-related cataract (ARC), including forty males (44 eyes) and thirty-five females (38 eyes). The age range was 41 - 85 years, the mean age was 67.97 years. There were 34 cortical cataracts, 22 nuclear cataracts and 26 subcapsular cataracts. Capsule specimens were divided into 4 groups: balanced salt solution (BSS) group I and group II (exposed to BSS for 1 minute), lidocaine group (exposed to preservative-free 1% lidocaine for 1 minute) and the control group. Specimens were stained with trypan blue and alizarin red. Photomicrographs of each capsule were taken to observe the viability of LECs and to count the number of necrosis LECs. The pathologic changes of LECs were evaluated by histological methods (11 LAC, 22 pieces) as well as transmission and scanning electron microscopes (5 LAC, 10 pieces). In the control and BSS group I (23 LAC), one half of each capsule specimen was used for the control group and the other half was used for BSS group I. In lidocaine group and BSS group II (43 LAC), one half of each capsule specimen was used for lidocaine group and the other half was used for BSS group II. RESULTS: The rate of necrosis LECs of the anterior capsules in the control group and BSS group I was (56.19 +/- 2.71)% and (57.23 +/- 1.98)%, respectively. The rate of necrosis LECs of the capsules in lidocaine group and BSS group II was (99.86 +/- 8.22)% and (57.64 +/- 7.00)%, respectively. Matching t-test showed that the rate of necrosis LECs in lidocaine group was greater than that in the BSS group II (t = 27.6781, P = 0.0000). There was no significant difference in the number of necrosis LECs between the control group and BSS group I (t = 2.0693, P = 0.0505). There was also no significant difference between males and females; between different cataract, types and between varying age groups (P > 0.05). After irrigated with lidocaine, LECs showed vacuoles and detached from the capsule, and many cavities appeared between the LECs and the capsule. The capsules of BSS and control group showed a normal layer of LECs attached to the capsule. Under transmission and scanning electron microscopes, in lidocaine group, the junction between the LECs and between the cells and the capsule were destroyed; many cells detached from the capsule and the rest arranged loosely. Some LECs dented and many vacuoles emerged, resulting in destruction of the cellular structures. CONCLUSION: Preservative-free 1% lidocaine may loose the junction between LECs and between the cells and the capsule, and also destroy cellular structures, resulting in degeneration and necrosis of the LECs. PMID- 19187660 TI - [bFGF siRNA expression plasmid inhibits growth of human LEC-B3 cells]. AB - OBJECTIVE: Basic fibroblast growth factor (bFGF) has been thought to play an important role during the development of posterior capsule opacification (PCO) by promoting the growth of lens epithelial cells (LECs). In the present study, we sought to explore the inhibition effect on LEC-B3 cells growth by plasmid-based RNA interference (RNAi) targeting bFGF. METHOD: It is a prospective study. LEC-B3 cells, an immortal human lens capsule epithelial cell line, were identified as human lens epithelium by immunohistochemical assay of alphaB-crystalline. Both bFGF and bFGF receptor 1 (FGFR1) mRNA expressions of LEC-B3 cells were determined by reverse transcription polymerase chain reaction (RT-PCR). Based on the identification of bFGF expression, plasmid-based RNA interference (RNAi) was used to inhibit bFGF mRNA expression of LEC-B3 cells by stable transfection. bFGF mRNA and protein levels were examined by real time PCR and western blot, respectively. Then the flow cytometry (FCM) was performed to evaluate proliferation cell nuclear antigen (PCNA) expression. PCNA level in bFGF-interfered LEC-B3 cells was compared to that of the control. P < 0.05 was regarded as statistically different. RESULTS: bFGF and FGFR1 mRNAs were abundantly expressed in LEC-B3 cells. bFGF mRNA and protein levels were both down-regulated by stable transfection of bFGF siRNA expression plasmid. PCNA expression was significantly decreased by inhibiting bFGF expression (t = -5.0011, P = 0.0005). CONCLUSIONS: bFGF and FGFR1 mRNA are extensively expressed in LEC-B3 cells. Plasmid-based RNAi targeting bFGF can lead to potent inhibition of LEC-B3 cells growth, which may play a part in dealing with the development of PCO. PMID- 19187661 TI - [Clinical analysis on the effects of laser-assisted subepithelial keratomileusis in treatment of super high myopia]. AB - OBJECTIVE: To assess the clinical efficacy and safety of LASEK in the treatment of super high myopia. METHODS: It was a retrospective case series study. Fifty six eyes of 33 cases with super high myopia were treated with LASEK. Mean spherical equivalent preoperative was (-11.70 +/- 1.59) D (range from -10.0 D to 15.13 D), mean ablation zone was (5.3 +/- 0.51) mm (range from 4.3 mm to 6.0 mm), mean ablation depth was (146.1 +/- 29.3) microm. The mean time of alcohol used treatment was (29.2 +/- 2.8) seconds. Postoperative follow-up assessments were performed at 1, 3, 6 and 12 months after surgery. Postoperative measurements included slit-lamp microscopy, manifest refraction, un-corrected visual acuity (UCVA), best corrected visual acuity (BCVA), intraocular pressure and haze, etc. The time of follow-up was more than one year. The results were analyzed with ANOVA by SPSS software. RESULTS: Fifty percent of eyes obtained UCVA of > or = 1.0, and 78.6% of eyes obtained UCVA > or = 0.6, at six months after operation. Forty-one percent eyes obtained UCVA of > or = 1.0, and 73.2% eyes obtained UCVA > or = 0.6 at one year postoperatively. The mean UCVA in Snellen chart was 0.85 +/- 0.20, 0.88 +/- 0.28, 0.84 +/- 0.26 and 0.83 +/- 0.28 at 1, 3, 6 and 12 months after operation, respectively. These data were transformed into lgMAR visual acuity and analyzed statistically. There was no significant statistically difference between UCVA at different follow-up periods (F = 0.749, P = 0.518). The postoperative UCVA compared to preoperative BCVA being the same/or improved in 71.4% eyes and decreased in 28.6% eyes at six months after operation; and was 64.3%, and 35.7% at one year after operation, respectively. The effective index was 0.994 and 0.992 at six and twelve months after operation, respectively. The postoperative BCVA compared to preoperative being the same (or improved) and decreased at six and twelve months after operation was 85.3% vs. 14.7%, and 84.8% vs. 15.2%, respectively. The mean BCVA in Snellen chart was 0.90 +/- 0.17, 0.94 +/- 0.24, 0.95 +/- 0.16 and 0.96 +/- 0.16 at 1, 3, 6 and 12 months after operation, respectively. These data were transformed into lgMAR visual acuity and analyzed statistically. There was no significant statistically difference between lgMAR BCVA at difference different follow-up periods (F = 1.567, P = 0.229). The safe index at 6 and 12 months were 1.009 and 1.010. The incidence of refraction regression at 3, 6 and 12 months after operation was 19.6%, 28.6% and 30.3%, respectively. Mean regressive diopter in regressed eyes at 3, 6 and 12 months after operation was (-1.20 +/- 1.08) D, (-1.23 +/- 0.75) D, (-1.49 +/- 1.04) D, respectively. Haze grade 0.5 was 21.4% at 3 months after operation; grade 0.5 and 1 were 8.9% and 5.4% at 6 months after operation, respectively, at 6 months after operation. Haze grade 0.5 was 12.5% at one year after operation. The incidence of high intraocular pressure induced by topical use of corticosteroid eye drop was 8.9%. CONCLUSIONS: LASEK is a safe and effective procedure for correction of super high myopia, though there is still some regression happened after operation and the intraocular pressure should be observed during the follow-up periods. PMID- 19187662 TI - [Viability of corneal epithelial cells and cleavage of corneal basement after ethanol effect]. AB - OBJECTIVE: To evaluate the viability of corneal epithelial cells and to determine the anatomic cleavage on the epithelial basement membrane after various exposure times to 20% ethanol during epithelial flap preparation in laser-assisted subepithelial keratectomy (LASEK) in cadaver eyes. METHODS: Six human cadaver eyes were exposed to 20% ethanol for 20, 30 and 40 seconds (2 eyes for each group), and another one eye was used as the control. PCNA staining was performed to determine the viability of corneal epithelial cells. Immunofluorescence staining using monoclonal antibodies against collagen VII, and immunohistological staining using monoclonal antibodies against laminin were performed to detect the anatomic location of the cleavage plane on the corneal epithelial flaps created by 20 seconds exposure to 20% ethanol in cadaver eyes. RESULTS: Hematoxylin and eosin staining of epithelial flaps revealed a coherent stratified epithelium. The PCNA positive rates of the epithelial cells in the flap decreased in the 20 second group, 30-second group and 40-second group successively. Immunohistological staining to laminin was patchy in the lifted flap and the remaining corneal basement membrane. Immunofluorescence to collagen VII, the main component of anchoring fibrils remained exclusively in the corneal bed. CONCLUSIONS: Viability of the epithelial flap decreased with longer time exposure to ethanol. The cleavage plane of the ethanol-treated corneal epithelial flap is located between the lamina lucida and the lamina densa of the basement membrane where laminin forms hemidesmosome. PMID- 19187663 TI - [Clinic observation of laser in situ keratomileusis for treatment of presbyopia with hypermetropia]. AB - OBJECTIVE: To investigate the effects of Laser in situ keratomileusis (LASIK) for treatment of presbyopia with hypermetropia. METHODS: 32 presbyopia patients (64 eyes) with hypermetropia were treated by LASIK and follow-up 1 year. The preoperative sphere was +0.75 to +3.00 D and cylinder was 0.00 to +1.75 D. For distance vision correction, the target refraction was 0.00 to -0.50 D in the dominant eye. And for near vision, the target refraction was -1.50 to -2.50 D in the non-dominant eye. The measurement including refraction, uncorrected visual acuity (UCVA), near vision, Q factor of cornea and contrast sensitivity were recorded. RESULTS: At 1 year after surgery a total of 53.1% (17/32) of all patients had binocular distance UCVA of 20/25 or better along with J3 or better near, and along with J4 or better near was 87.5% (28/32). The postoperative corneal refractive power was significant higher than preoperative [3 mm: (44.12 +/- 1.38) D, 5 mm: (43.42 +/- 1.53) D] and Q factor became more negative. At 1 year 87.5% (28/32) and 100.0% (32/32) of the dominant eyes treated for distance had a manifest refractive spherical equivalent within +/- 0.50 and +/- 1.00 D of intended correction, respectively; of the non-dominant eyes treated for near, 56.2% (18/32) and 90.6% (29/32) had a manifest refractive spherical equivalent within +/- 0.50 and +/- 1.00 D of intended correction respectively. The difference of sensitivity was no statistically significant between preoperative and 6 months after surgery. There were 15.6% of the eyes had dry eye syndrome and 84.4% of the patients were satisfied with LASIK at 1 year after surgery. CONCLUSIONS: LASIK was a effect surgery for treatment of presbyopia with hypermetropia. PMID- 19187664 TI - [Comparative study of gentamycin and povidone-iodine on eliminating the conjunctival bacteria]. AB - OBJECTIVE: To study the efficacy of gentamycin, 0.5% povidone-iodine and 5.0% povidone-iodine in eliminating rate of positive conjunctival bacteria cultures and aqueous humor contamination, and to observe their adverse effects. METHODS: It was a comparative study. Three hundred patients (300 eyes) were randomized to three groups: gentamycin group, 0.5% povidone-iodine group and 5.0% povidone iodine group. All patients received 1-day (6 times) topical application of 0.3% ofloxacin before surgery. Approximately 5 minutes before surgery, the conjunctival sac was irrigated with 5 - 8 ml gentamycin, 0.5% povidone-iodine or 5.0% povidone-iodine, correspondingly. Conjunctival cultures were obtained at the following time points: before receiving any topical medications; after application of 1-day topical 0.3% ofloxacin, before irrigating the conjunctival sac; immediately before surgery approximately 5 minutes after conjunctival sac irrigation and at the end of the surgery. Anterior chamber aspirates were obtained at the beginning of the surgery. Bacteria isolated were identified and antibiotic susceptibility was determined. The rate of positive conjunctival cultures among each group was compared and significance of the difference was tested with the Pearson chi-square test. RESULTS: Rate of positive conjunctival cultures before the conjunctival irrigation was 79.7%, 76.6% and 77.8% in the gentamycin group, 0.5% povidone-iodine group and 5.0% povidone-iodine group, respectively. The difference between these three groups was not statistically significant (chi2 = 0.28, P = 0.86). At the end of surgery, the rate of positive conjunctival cultures in these three groups was 11.7%, 8.3% and 6.3%, respectively. The difference between these three groups was also not statistically significant (chi2 = 1.74, P = 0.41). The average rate of anterior aqueous contamination was 4.9% and there was no significant difference among these three groups (P > 0.05). CONCLUSIONS: Both 0.5% povidone-iodine and 5.0% povidone-iodine using for irrigating conjunctival sac are safe. Gentamycin, 0.5% povidone-iodine and 5.0% povidone-iodine have similar effects on the reduction of the bacteria in conjunctival sac. PMID- 19187665 TI - [Variations of the microstructure in human limbus by confocal microscopy in vivo]. AB - OBJECTIVE: To demonstrate the variations in the niche of limbal stem cells and the limbal basal epithelial cells in vivo by confocal microscopy. METHODS: One hundred and twenty eyes of 120 healthy subjects were enrolled in this study after a routine slit-lamp examination. They were divided into four groups according to the age: group A (0-), group B (20-), group C (40-) and group D (60 - 79). Confocal microscopic examination in vivo was performed and the images of the inferior limbus were recorded. The morphologic features of each group were analyzed and the average size of limbal basal epithelial cells of each group were measured. RESULTS: The morphologic features of limbus varied among groups. 96.7% (29/30) of group A showed clear Vogt Palisades, while 3.3% (1/30) showed no Vogt Palisades. 96.7% (29/30) of group B also showed clear Vogt Palisades, while 3.3% (1/30) showed no Vogt Palisades.70.0% (21/30) of group C showed clear Vogt Palisades, while 13.3% (4/30) showed atrophic Vogt Palisades and 16.7% (5/30) showed no Vogt Palisades at all. 33.3% (10/30) of group C showed clear Vogt Palisades, while 10.0% (3/30) showed atrophic Vogt Palisades and 56.7% (17/30) showed no Vogt Palisades at all. The average size of limbal basal epithelial cells in each group was (9.89 +/- 1.12), (10.65 +/- 1.45), (10.70 +/- 1.39) and (12.22 +/- 1.42) microm in sequence. CONCLUSIONS: There is variations in the microenvironment of niche in human limbus. Meanwhile, limbal basal epithelial cells showed different quantity and cell size, demonstrating varied proliferative potential. PMID- 19187666 TI - [Long-term follow-up and prognostic analysis on the silicone oil-dependent eyes]. AB - OBJECTIVE: To study on the outcome of the eyes with silicone oil tamponade retrospectively, and to observe ocular complications and prognosis after long term intraocular silicone oil tamponade. METHODS: Retrospective observational case series. 36 patients (37 eyes) with silicone oil tamponade for over 2 years without removal were analyzed from 1992 to 2005 in our hospital. The patients were 30 males (31 eyes) and 6 females (6 eyes); age ranged from one year and 2 months to 73 years old. The lasted time of silicone oil tamponade were from 2 to 13 years. 37 eyes were all open ocular trauma, including traumatic eyeball rupture 24 eyes, intraocular foreign body 5 eyes, ocular perforating injury 3 eyes and penetrating injury 5 eyes. Vitrectomy, retinotomy and retinectomy, retinal photocoagulation and silicone oil tamponade were performed in different patients. RESULTS: Complete retinal attachment was seen in 13 eyes (35.1%); partly retinal attachment was accomplished in 11 eyes; retinal detachment with silicone oil tamponade was revealed in 7 eyes. Visual acuity in 9 eyes was better than 0.01 (24.3%), the best vision acuity was 0.1 among all of patients. The complications in the long-term follow-up included: cornea band-shaped degeneration 13 eyes (35.1%); complete corneal opacity 6 eyes; silicone emulsification 17 eyes (45.9%); secondary glaucoma 2 eyes; cataract 4 eyes, without obvious complications 8 eyes (21.6%). CONCLUSIONS: Long-term intraocular silicone oil tamponade will be adaptive and rational in serious ocular trauma if no other better treatments can maintain retinal attachment and the appearance of eyeball. PMID- 19187667 TI - [Study on the setup of a new technique of tissue and cell culture for vitreous fluid of vitrectomy]. AB - OBJECTIVE: To setup a new technique of tissue and cell culture for vitreous aspirates. METHODS: Experiment study. Specimens used for supporting new culture technique were selected based on random digit table. Thirty cases with rhegmatogenous retinal detachment (RRD) and forty-eight with proliferative diabetic retinopathy (PDR), which undergoing primary pars plana vitrectomy, were selected randomly and included in the study. After being antiphase stained with fluorescein-natrium (0.5%) and digested with hyaluronidase (10(5) U/L) combined with collagenase I (10(6) U/L) for removing vitreous gel, sediment of vitreous fluid after centrifugation were inoculated into standard culturing bottle with which polylysine (0.01%) was pre-set. The bottle which contained F12 medium with 30% fetal bovine serum was placed upside down for 24 hours and consecutively upside for 6 days. During which, F12 medium was replaced once in half volume, and cell growth along the edge of sedimentary membrane was observed at time of the 3rd and the 6th day after upside culture. RESULTS: Under condition of pre-setting by polylysine (0.01%) and being placed upside down for 24 hours, pieces from vitreous fluids could adhere to the bottom of bottle in a way of semi-xerosis with adherence rate of 100% (78/78). No bacteria, fungus and mycoplasma contamination was found within 7 days. Antiphase stained with fluorescein-natrium (0.5%) and digested with hyaluronidase (10(5) U/L) combined with collagenase I (10(6) U/L) for 30 minutes, vitreous gel in 78 specimens could be digested (78/78). Cell emigration could be found in edge area of some pieces of vitreous fluid and cell growth as well as proliferation was shown. In 30 specimens of RRD, cell growth rate were 43.33% (13/30). In 48 specimens of PDR, cell growth rate were 37.50% (18/48). Concerning PDR phase V (PDR-V), cell growth rate reach 41.67% (10/24). CONCLUSIONS: Enzymolysis with upside down and semi-xerosis could ensure good adherence of membrane, moreover, no contamination and obvious cell growth could be found within short-term culture. These suggested that a new technique for judging viability of cell from proliferative membrane and predicting recurrent risk after surgery of proliferative ocular fundus disease could be expected. PMID- 19187677 TI - [Clinical research of intraoperative floppy iris syndrome during operation]. AB - Intraoperative floppy iris syndrome (IFIS) has been recently identified as a new small pupil syndrome during phacoemulsification. This syndrome is characterized by three intraoperative features: a flaccid iris stroma that undulates and bellows in response to intraocular fluid currents; a propensity for the floppy iris stroma to prolapse toward the tip of phacoemulsification and side-port incisions despite proper wound construction; and progressive intraoperative pupil constriction despite standard preventive preoperative pharmacologic measures designed to prevent this. It is now mostly considered that IFIS is associated with the use of tamsolusin, a highly selective alpha-1A receptor antagonist for the treatment of benign prostatic hypertrophy. It is recommended that a careful history of the use of alpha-1 blocking agents be taken before cataract surgery to anticipate the occurrence of IFIS. A combination of strategies could decrease the complications of IFIS. These procedures include preoperative use of atropine, intracameral injection of dilute phenylephrine or epinephrine, the use of super cohesive ophthalmic viscosurgical devices, lower phacoemulsification vacuum and aspiration settings and various iris hooks or pupil dilators. PMID- 19187679 TI - [Endothelial keratoplasty]. AB - Endothelial keratoplasty (EK) is a new surgical method in treatment of corneal endothelial keratopathy, which retains healthy portions of a patient's cornea while replacing diseased endothelium with healthy donor tissue. In the past decade EK has been used increasingly and has shown better results over standard penetrating keratoplasty. The emergence and improvement of EK demonstrate the convert of corneal transplantation to refined and refractive surgery. EK, which has favorable refractive outcomes and potential low rejection rate, opens a new era for the treatment of endothelial dysfunction. However, there are still some aspects requiring to be studied, such as the methods of implantation to decrease graft dislocation rate and endothelial cell loss, use of femtosecond laser in EK to improve refractive errors, and the long-term result of immune rejection rate. PMID- 19187678 TI - [Present research of hot shock protein and cataract occurrence]. AB - The hot shock proteins (HSP) were discovered by Ritossa in the fruit fly salivary gland after heat stimulation, giving the name HSP. In following comprehensive studies, it was recovered that HSP was a group of highly conservative cell stress proteins in all organisms. HSP are expressed under many stress conditions, e.g. in the presence of high temperature, heavy metal, oxygen deficit, ethyl alcohol and viral or bacteria infections, etc. HSP, as the molecular companion, play a vital role in the folding, stability and synthesis of the protein. The lens opacification (cataract) is one of most common causes of blindness in the world. Various studies indicate that the occurrence of cataract is directly related to free radical production, oxidized damage, changes of the proportion of lens protein and apoptosis of lens capsule epithelial cells, etc. Recent studies indicated that the lens expresses a series of HSP. They play a key role in maintaining the stability and transparency of the lens. This article is a summary of the relationship between the HSP and the occurrence of cataract. PMID- 19187682 TI - [Efficiency versus quality in the NHS, in Portugal: methodologies for evaluation]. AB - To proceed to the evaluation of the efficiency and quality in the NHS, based in methodologies of evaluation of management, indicators of benchmarking and indicators of process and outcome. The 1980 and 1990 decades have seen the proliferation of all forms of process indicators as a way to control health services. It is not a coincidence that the increase in managed care has been accompanied by an explosion of process indicators, as it has happened in the health system of the USA. More recently the attention has turned away from measures of performance, which measure the process (what has been done) to those which measure outcomes (what was the result). Quality indicators have been developed in Europe, first to be used in hospitals, but also to be used in primary health care. Conceptually the justification for the introduction of process indicators comes from the principle that their use will reinforce a modification in the quality of the proceedings, which will give origin to better outcomes as well at population level, as resource saving. Outcome indicators compared with process indicators in health care shows that process indicators have the advantage of being more sensitive than outcome indicators to differences in the quality. Optimizing health care quality has the objective of establishing a quantitative relationship between the quality of the health services and cost effectiveness. To identify quality indicators and benchmarking and to implement plans to measure the quality of health care. In a study made in a group of senior GP, in the UK, with the objective of determining which process indicators better reflect the quality of the services in primary health care services a Delphi method was used. Only seven indicators were chosen by 75% of the respondents: the percentage of eligible patients receiving cervical screening; the percentage of generic prescribing; the percentage of eligible patients receiving childhood immunization; the percentage of eligible patients receiving influenza vaccinations; ability to see GP within 48 hours; percentage prescribing antibacterial drugs; primary care management (diabetes and asthma). The main characteristics of health indicators are: acceptability--The acceptability of the data collected using a measure will depend upon the extent to which the findings are acceptable to both those being assessed and those undertaking the assessment; feasibility--information about the quality of services is often driven by data availability rather than by epidemiological and clinical considerations. Quality measurement cannot be achieved without accurate and consistent information systems; reliability--indicators should be used to compare organisations/practitioners with similar organisations/practitioners; sensitivity to change--quality measures must be capable of detecting changes in quality of care in order to discriminate between and within subjects; validity--there has been little methodological scrutiny of the validity of consensus methods. Outcome indicators are not good performance indicators in health care. Which causes the variation in outcomes between deliverers of primary health care services are the observed differences due to differences in users, due to age, sex, co-morbidity, severity and socio-economic situation. The Medical Outcomes Study, published in 1989, has brought, for the first time, subjective indicators, based in the evaluation of users, as an important outcome indicator. Clinical indicators are those that are more associated with the outcomes. A few studies exist of the effects of management indicators in outcomes. Several indicators, however, reflect norms related with the local of work. The use of a Composite Indicator presents advantages. In England it has been used a Composite Indicator of process indicators in 302 organizations of primary health care, in 2001-2002. This study has used a mathematical model to select the best indicators which allow the evaluation of performance. It has concluded that the use of a Composite Indicator is of easy construction, interpretation, and acceptable and that has validity. Giraldes (2007) has done an evaluation of health centres in a perspective of management and quality of deliver using a Composite Indicator of Efficiency and Quality. It includes the efficiency indicators concerned with the main activities of the health centre, preventive activities, curative activities and drugs, by main pharmaco therapeutic groups, and auxiliary means of diagnosis (analysis, X Ray, ecographies and CAT by user, weighted according to the relevance of the expenditure in total expenditure). The Composite Quality Indicator includes 12 performance and 5 outcome indicators. From the 10 best health centres in an efficiency and quality perspective 3 are from the Porto Sub-Region (Negrelos, Rebordosa and Paredes) and 2 from the Braga Sub-Region (Vila Verde and Vila Nova de Famalicao I), Leiria (Pedrogao Grande and Batalha), and Vila Real (Mesao Frio and Sabrosa), while 1 belongs to the Aveiro Sub-Region (Sever do Vouga). The more efficient health centres are from the Aveiro Sub-Region, followed by Braga, Porto, and Lisboa. Sub-Regions with very similar values. Giraldes (2007) has made an evaluation of the hospital expenditure by user in an efficiency perspective and to evaluate the quality of the health system process indicators and outcome indicators. In an efficiency perspective the concept of technical efficiency has been chosen, and a correction has been made, as well, in what concerns a case-mix index (CMI). The indicators have been calculated by user in what concerns the main hospital activities (the expenditure in inpatient care by treated patient, in day hospital by treated patient, in outpatient care by consultation, etc.) and as well the auxiliary sections of clinic support and the hotel support services. All the indicators have been corrected according to the relevance of its expenditure in total expenditure. In a quality perspective two types of indicators have been considered; process indicators and outcome indicators. Process indicators, as the percentage of surgeries in ambulatory care, the percentage of cesareans in total deliveries and the rate of autopsy. The outcome indicator number of episodes of inpatient care due to surgery infection in total days of inpatient care. Those indicators have been aggregated, by a mean. The Composite Indicator of Efficiency and Quality is the mean of the Composite Indicator of Efficiency and the Composite Indicator of Quality, having this one been converted in inverse base. PMID- 19187683 TI - [Use of granulocyte growth factors: recommendations of the Portuguese Society of Hematology]. AB - The administration of cytotoxic chemotherapy may be complicated by the emergence of neutropenia and febrile neutropenia, frequently determining hospital admission and intravenous treatment with broad spectrum antibiotics. Frequently, it is necessary to reduce the dose or to delay the administration of the cytotoxic drugs reducing the relative dose intensity of the chemotherapy regimen. Granulocyte growth factors stimulate the proliferation and differentiation of neutrophils and reduce the number of days of severe neutropenia and febrile neutropenia associated with cytotoxic chemotherapy. They are also indicated for the collection of hematopoietic progenitors for autologous and allogeneic transplantation, as well as in non malignant diseases associated with chronic neutropenia. This article reviews the evidence supporting the use of granulocyte growth factors in Hematology. PMID- 19187684 TI - Indications for upper gastrointestinal endoscopy. The accuracy of the American Society for Gastrointestinal Endoscopy guidelines in a Portuguese hospital. AB - INTRODUCTION: The appropriateness of the indications for upper gastro-intestinal endoscopy (EGD) is crucial in assessing quality in endoscopy units, improving cost-effectiveness and providing better patient care. Using the American Society for Gastrointestinal Endoscopy (ASGE) guidelines, the aim of this study was to evaluate the appropriate use of EGD and to measure the local accuracy of these guidelines. PATIENTS AND METHODS: Over a two year period information was gathered on 2305 consecutive patients--1146 (50% males)--of whom had an EGD performed at our unit. Patients were referred for EGD by other physicians of the hospital staff or through the gastroenterology out-patient clinic. The appropriateness, sensitivity, specificity and predictive value of the ASGE guidelines were established using EGD diagnosis as gold standard. Atrophic gastritis and hiatus hernia were considered irrelevant diagnosis. RESULTS: According to the ASGE guidelines the rate of inappropriateness was 20%. No lesions or irrelevant lesions were found in 30.6% EGD. Appropriately indicated endoscopies disclosed significantly more clinically relevant findings (71.3%) than endoscopies performed with indications that were not ASGE listed (61.7%) - p < 0.01, OR = 1.55, 95% CI (1.24-1.92) but no significant difference was found between appropriateness and inappropriateness in patients with a diagnosis of gastric cancer: p = 0.21, OR = 1.53, 95% CI (0.75-3.21). The sensitivity of the ASGE criteria was 82.1%, the specificity 25.2%, the positive predictive value 71% and the negative predictive value 38.1%. Gastric cancer was found in 10 (0.4%) of the patients not appropriately indicated. CONCLUSIONS: In this Portuguese population sample, the accuracy of the ASGE guidelines is too low to be confidently acceptable. This suggests that, in Portugal, a country with a high prevalence of gastric cancer, wider criteria must be applied, if useful local guidelines for appropriate referrals are expected. PMID- 19187686 TI - [Operationalization to Portugal: Beers criteria of inappropriate medication use in the elderly]. AB - INTRODUCTION: Elderly drug therapy needs special care considering physiopathological alterations of this age group that increase the risk of adverse drug events occurrence and due to the high number of drugs used. Several tools have been created, as tables of drugs and groups of drugs to be avoided in patients 65 years old and over. Beers Criteria of 2002 update is the most used tool. GOALS: To operationalize Beers Criteria to be used by health care professionals in Portugal. METHODS: Analysis of the drugs and therapeutic classes included into the Beers Criteria approved for market in Portugal and the identification of other approved substances included in the pharmacological classes mentioned in Beers Criteria. The two tables in the Beers criteria were adapted to substances approved in Portugal, even including other marketed active substances belonging to therapeutic classes mentioned in Beers criteria. Levels of inappropriateness and the potential effects after their use in elderly were also included. In Beers table 1, 34 included substances don't have approval in Portugal, and 4 have different denominations. In Beers table 2, three tricyclic antidepressants, 12 typical antipsychotics, 5 doses for short-action benzodiazepines, and 28 NSAIDs (being two coxibs) were included. CONCLUSIONS: The Portuguese operationalization of the Beers Criteria allows the creation of a tool that helps prescribers to choose drugs and doses for a safer prescription to the elderly. These adapted tables allow benchmarking among studies assessing inappropriateness of use of drugs in different countries using Beers Criteria. PMID- 19187685 TI - [Influence of the APOE genotypes in some atherosclerotic risk factors]. AB - The aim of this work was to study the distribution of apolipoprotein E (APOE) genotypes and their association with some atherosclerotic risk factors, all of them modifiable: total, HDL and LDL cholesterol, triglycerides, systolic and diastolic blood pressure, BMI, waist circumference and smoking. The sample population was constituted of 672 healthy subjects recruited in the Lisbon area. Lipids were quantified by usual automatic enzymatic methods and the APOE genotypes performed in accordance with Hixson and Vernier. Blood pressure measurement and hypertension classification followed international specifications. The frequency distribution of APOE alleles was: epsilon2 = 6.4%, epsilon3 = 83.6% and epsilon4 = 10.0% and the more prevalent genotypes were epsilon2/epsilon3, epsilon3/epsilon3 and epsilon3/epsilon4 respectively 11.0%, 70.1% and 16.1%. We could only observe associations among the most prevalent genotypes and lipids, always statistically significant, specially when the epsilon4 allele was present which was even proved by an higher prevalence of epsilon4 in dyslipidemic subjects with the only exception of those with low HDL-c values. A stronger intervention in the epsilon4 carriers is so recommended through appropriate intervention measures on the connected modifiable risk factors. PMID- 19187687 TI - [Medicines and the body: what is it consumers think and know about medicines]. AB - Little is known about what patients think of the medicines they take. Such knowledge could be of importance for health results. In a primary care setting, the knowledge about pharmacokinetics, pharmacodynamics and about how medicines work in the body were addressed through a validated questionnaire for self fulfillment, in anonymity, to be returned by mail. 272 questionnaires were received representing the population in study in gender and age. As main results knowledge about pharmacokinetics, pharmacodynamics is scarce and there is good knowledge about drug adverse events even though patients don't know how it is medicines work in the body and show very good knowledge about safety medicine's issues. As main conclusions there is little knowledge about how medicines work in the body, much information work being needed to upgrade what patients know in order to ameliorate health results, when medicines are needed. PMID- 19187688 TI - [A thousand total colonoscopies: what is the relationship between distal and proximal findings?]. AB - BACKGROUND: Flexible sigmoidoscopy is indicated for colorectal cancer screening. The decision about who needs total colonoscopy based on distal findings is still controversial because of the uncertainty of the associations between distal and proximal findings. AIM: The purpose of the study was to characterize distal findings in patients with total colonoscopy, to investigate its importance as markers of advanced proximal lesions and to evaluate the usefulness of a clinical Predictive Index, already published in the literature, in the identification of these lesions. METHODS: Retrospective analysis of the patients submitted to total colonoscopy between January 2006 and February 2007, with selection of 1000 consecutive cases with reference to polyps. We analysed demographic data, indication for the exam and morphological and histological characteristics of the polyps. Advanced lesion was defined as any adenoma larger than 10 mm or any polyp with villous characteristics, high grade dysplasia or cancer. The Predictive Index was obtained through the assignment of points to 3 categories: sex, age and distal findings, which result in 3 groups: low, intermediate and high risk. RESULTS: The mean age of patients was 64,69 years and 65,1% were male. Distal and proximal polyps were identified in 829 (82,9%) and 369 (36,9%) patients, respectively. Advanced distal lesion was found in 342 patients (34,2%) and advanced proximal lesion in 98 (9,8%). 587 patients (58,7%) were in the high risk group. In the group of patients with advanced proximal lesion, a third presented low and intermediate risk, 52% had no distal polyps, 88,7% had less than three distal polyps and 71,4% had no advanced distal lesion. Sensitivity values for these four categories ranged between 11,2% and 66,6%. CONCLUSION: If the decision to perform total colonoscopy is based on distal colonic findings or on the Predictive Index, the ability to identify advanced proximal lesion is markedly reduced, endangering the aim of a screening program. PMID- 19187689 TI - [Children's oral hygiene: healthy mouth, families supervise?]. AB - Dental caries results from a complex interaction between the host and environmental factors and it is an important public health issue. AIMS: To determine the prevalence of dental caries in a school population of six and 12 years old children from Leiria; to establish a relationship between dental caries and related known risk factors; parental dental health care, parental control of their children oral hygiene and to compare our results with the National Dental Care Study of 1999. METHODS: Descriptive and statistical analysis based on a questionnaire and dental examination. RESULTS: In our sample of 248 children, 43% were six years old and 57% 12 years old; 52% were female and 72% reside in a suburban area. The prevalence of dental caries was 42% (48% for the six year and 33% for the 12 years old groups. Dental caries were more prevalent in males (p = 0, 01) and 25% of these had three or more caries. Analysis of the questionnaire revealed that 87% of children brushed their teeth daily, findings that were unrelated to gender, age or residential area. Of this group, 68% brushed their teeth twice or more daily (p = 0,008). This routine was commenced in 32% of children prior to the age of three and these had less dental caries (p = 0,022). With regard to the children's dietary habits, we found them to be similar in both groups. The children who did not sweeten their milk (23 and 24% of the six and 12 years group, respectively) had fewer dental caries (p = 0,031). Dental treatment was required in 53% of 12 year old group and in 41% of six year old group. Dental caries was also less prevalent in children who also had taken fluoride (p = 0,045). We found a significant statistical relationship between parental and their children's dietary habits (p = 0,000). Parents who brushed their teeth twice daily had children with similar oral hygiene habits and had fewer caries (p = 0,002). Parents who oversee their children's dental care (52%) include the group of parents and children that brush their teeth at least twice daily (p = 0,003). CONCLUSIONS: Dental caries was more prevalent in the six year old group with a male predominance. Children who brushed their teeth twice a day had less caries and the 12 year old group had more visits to the dentist. To ensure adequate oral hygiene habits it is important that parents supervise their children' habits. PMID- 19187690 TI - [Neurocognitive dysfunction after valve surgery]. AB - Neurocognitive dysfunction is, nowadays, reported as one of the most frequent complications of cardiac surgery, with documented potential to have a negative impact on quality of life. Notwithstanding, the cognitive evaluation is almost exclusively restricted to research contexts, being depreciated in clinical settings. Cognitive functioning changes have mostly been investigated in patients submitted to coronary artery bypass grafting, with data reporting to valvular surgery still being extremely rare. In spite of the dramatic reduction of rheumatic valvular pathology in developed countries, the burden of degenerative diseases owed to prolonged life expectancy keep the valvular heart diseases as a major Public Health problem. Beyond the intraoperative damage, which [corrected] is believed to occur in both surgeries, in valvular surgery the deficits might turn out to be the result of cumulative lesions caused by microemboli originating from prosthetic cardiac valves. Etiology of cerebral injury related to cardiac surgery is not completely elucidated, probably representing a complex interaction between cerebral microemboli, global cerebral hypoperfusion, inflammation and genetic susceptibility. The exact identification of the involved mechanisms is still a great challenge. The available data concerning cognitive performance after valvular surgery point out that this intervention appears to be more harmful towards cognition than the coronary artery bypass grafting, even when considering biological valve replacement. Thus, the type of valve prosthesis appears not to be the prevailing factor in cognitive complains maintenance, with age and type of valvular intervention being the most influent factors on long term prognosis. A notable variability between results of different studies may arise from diverse investigational methodologies and from factors related with difficulties inherent to the cognitive assessment, such as the diverse methods used for evaluation and the presence of a remarkable number of confounding factors. Investigation on valvular surgery neurocognitive effects is still in a very incipient period, being crucial to accurately establish the exact influence of the specific variables of this particular patient's group, such as the type of surgery and valve prosthesis. Furthermore, the clarification of the pathophysiological mechanisms subjacent to neurocognitive dysfunction might represent an important step to look for potential preventive or therapeutic strategies that could ameliorate brain function after cardiac surgery. PMID- 19187691 TI - [Hysteroscopic diagnostic accuracy in post-menopausal bleeding]. AB - OBJECTIVE: To evaluate the diagnostic accuracy of hysteroscopy at our unit for postmenopausal bleeding, specially concerning proliferative lesions. METHODS: We analyzed the results of 335 hysteroscopies that were done during the last 8 years in our hysteroscopic unit and we compared the hysteroscopic diagnosis with the biopsy result done during the procedure. Data were group according to pathologic findings in three groups normal, proliferative lesions and benign lesions. Data were analyzed by direct comparison. Sensitivity and specificity were calculated, Kappa index was used to access the inter-rater reliability, and likelihood ratio and the post test probability were used to analyze the true value of hysteroscopy. RESULTS: Women were aged between 36 and 88 years, with an average of 61,5. Histological diagnosis was atrophy in 42,1% patients, polyps in 43,3%, sub-mucous fibromioma in 5%, hyperplasic lesions in 9,6%, half of them being carcinomas. Overall Kappa index for the 3 groups was 0,831 which is in line with excellent agreement. Concerning proliferative lesions (hyperplasia + carcinoma vs carcinoma alone) and comparing to histology, sensitivity was 78,1% vs 81,3%; specificity 95,7% vs 98,7%. The positive likelihood ratio was 18,2 vs 64,8 and the negative likelihood ratio was 0,23 vs 0,19. The probability post positive test was 66% vs 76% and the probability post negative test was 2,4% vs 0,95%. No cases of carcinoma were identified among the 129 women diagnosed as having atrophy. However 3,9% of all the lesions regarded as being polyps at the hysteroscopy proved to be proliferative lesions at hysteroscopy. CONCLUSION: Hysteroscopy at our unit is a highly accurate procedure concerning post menopausal endometrial bleeding, with its results being in line with the literature. Diagnosing atrophy or excluding a proliferative lesion by the observer was highly predictive of a negative carcinoma in the histology. Using this argument whenever a proliferative lesion was excluded, only 1 in 302 hysteroscopies hid a carcinoma. Polyps should be regarded as possible proliferative lesions. Despite this result we believe a biopsy should always be undertaken no matter the observer's diagnosis. PMID- 19187692 TI - [The role of adipose tissue and macrophages in chronic inflammation associated with obesity: clinical implications]. AB - In the last decades, life style modifications particularly those related to food patterns and food choices, gave risen to a worldwide pandemic disease with no precedent in Human History: obesity. Energy storing and free fatty acids production were the only adipose tissue functions. However, it has been well established other adipose tissue functions like low grade pro-inflammatory molecules production (cytokines, adipokines and chemotactic factors) evolved in obese inflammatory condition. In this inflammatory state, adipocytes active role is inflammatory mediators (adipokines) production and and cell to cell interaction with resident macrophages. Adipose tissue low vascularization is even lower in the obese; so, hypoxia can be a critical factor in inflammatory obese state manifestation. Adipose tissue cytokines production and pre-adipocyte conversion into macrophage results in adipose tissue and macrophages interactions. One of the mononuclear phagocytic system components, macrophage, has an important role in obese-related inflammatory state. These cells have been found to be increased in number and shape proportional to Body Mass Index, rising to up to 60% of total adipose tissue components. The macrophages proportional accumulation could lead to an increase in pro-inflammatory molecules expression and contribute to the inflammatory state in a significant way. The reversion of the low grade inflammation and the reduction of risk factors in obese individuals seem to occur when a reduction in Body Mass Index is achieved and loss of adipose tissue is observed. We performed a critical review of last five years literature in order to better describe the impact of adipose tissue and macrophages interactions in low grade chronic inflammatory condition in the obese and their role in comorbidities pathogenesis. PMID- 19187693 TI - [Chemokine receptors and its importance in the replication cycle of human immunodeficiency virus: clinical and therapeutic implications]. AB - Human immunodeficiency virus (HIV) infection of host cells begins with binding of viral envelope (Env) surface (SU) glycoprotein to specific receptors present at plasma cell membrane. One of these receptors is the CD4 molecule which can be found namely in T-helper lymphocytes (T-CD4+), macrophages and dendritic cells. Binding of SU glycoprotein to CD4 molecule, enables HIV to adsorb to target cell membrane and also initiates conformational changes in SU glycoprotein that enable it to bind to a second receptor (coreceptor). This coreceptor belongs to a family of plasma cell membrane molecules that acts, in vivo, as chemokine receptors. The SU glycoprotein binding to coreceptor molecule triggers further conformational changes and consequently the exposure of a hydrophobic region of viral envelope transmembrane glycoprotein (TM), named fusion peptide that ultimately leads to viral envelope fusion with target cell membrane. Nowadays, 19 of these chemokine receptors have been thus considered, in vitro, as coreceptors for HIV. Interestingly, despite this extensive range of potential coreceptors, only CCR5 and CXCR4 seem to be relevant in HIV transmission and in the pathogenesis of HIV infection. Identification of cell surface coreceptors, specific for HIV envelope SU glycoprotein, has provided an elucidative explanation for molecular mechanisms involved in viral cell tropism and pathogenesis. Furthermore, the recognition of a coreceptor-mediated HIV's entry has also provided novel viral and cellular targets for antiretroviral intervention. During the last few years, the inhibition of HIV entry has become an incontestable target for anti-HIV drug discovery. Enfuvirtide is one example of these new antiretroviral molecules. It is the only member of fusion inhibitors targeting fusion peptide region, which prevents HIV entry by blocking the TM-mediated fusion between viral envelope and plasma cell membrane. More recently, CCR5-specific antagonists have been described, including monoclonal antibodies, modified chemokines and more importantly small-molecules inhibitors, such as maraviroc and vicriviroc. These drugs prevent SU glycoprotein binding to CCR5 coreceptor, and thus inhibiting HIV entry into target cell. This review will focus on the influence of coreceptor engagement, by HIV Env glycoproteins, in viral replication cycle and the importance of targeting its coreceptor function, by specific inhibitors, as a new and promising class of antiretrovirals. PMID- 19187694 TI - [Osteonecrosis of the mandible induced by intravenous bisphosphonates in oncological patients]. AB - Bisphosphonates are synthetic analogs of inorganic pyrophosphate, which are not only potent inhibitors of osteoclast-mediated bone resorption, but also present an important inhibitory activity of angiogenesis. Bisphosphonates become, since 1996, the standard of care in the management of patients with osteoporoses as well as bone metabolism alterations associated with neoplasias. In 2003 the first case-reports of bisphosphonate induced osteonecrosis of the jaws were published. The authors report 2 new cases of patients with osteonecrosis induced by bisphosphonates. Case 1: Seventy one years-old male patient, undergoing standard antineoplastic therapy and bisphosphonates due to multiple myeloma 4 years ago was referred to an oral surgeon because of bone exposure, after dental extraction. Case 2: Seventy years-old female patient undergoing treatment with bisphosphonates for adenocarcinoma of the breast was referred to an appointment of oral surgery due to bony exposure. The increasing incidence of this new entity, its physiopathological mechanisms and the nonexistence of consensual and evidence-based treatments, calls for a special attention on prevention, with special emphasis on a careful clinical examination of the oral cavity previously to the establishment of a therapeutic approach with intravenous bisphosphonates in oncological patients. PMID- 19187695 TI - [Eosinophilic gastroenteritis: a rare case of dyspepsia]. AB - Eosinophilic gastroenteritis is a rare and heterogeneous disease that is probably misdiagnosed in clinical practice because it meets a high degree of suspicion, and an endoscopy with biopsy is needed for diagnosis. The authors describe, with a brief thematic review, a case of eosinophilic gastroenteritis presenting as dyspepsia syndrome and mild peripheral eosinophilia. PMID- 19187696 TI - [Paediatric Burkitt lymphoma presenting as acute pancreatitis]. AB - Pancreatitis in children and adolescents is uncommon and its causes are more varied in this age group than in adults. A tumoral aetiology is particularly rare. We present the case of a 13-year-old boy who was admitted to our Intensive Care Unit with the diagnosis of acute pancreatitis, bilateral pleural effusion and ascites. Serial sonographic and computed tomography evaluations were suggestive of an infiltrative process of the gastric wall. Endoscopy showed an infiltrative tumor of the gastric mucosa and duodenum with a giant ulcer. Biopsies were compatible with Burkitt Lymphoma confirmed by ascitic fluid cytology and cytometry. Because of severe progressive cholestasis a temporary biliary stent was placed in the common bile duct. After staging (stage III), the patient was treated according to FAB LMB 96 chemotherapy protocol, achieving complete remission. Acute pancreatitis and cholestasis are rare presentations of lymphoma in the paediatric age group. PMID- 19187698 TI - Zolpidem extended release for insomnia in older adults. PMID- 19187697 TI - Aripiprazole for psychosis and agitation in Alzheimer's dementia. PMID- 19187699 TI - A multinational trial of counseling for Alzheimer's disease caregivers. PMID- 19187700 TI - Effects of stopping antipsychotics in nursing home patients with dementia. PMID- 19187701 TI - Activities for behavioral problems in patients with dementia. PMID- 19187702 TI - Sexual functioning in older adults. AB - This article reviews recent medical and social science literature on sexual functioning in older adults. We provide a broad definition of sexual functioning that includes a range of solo and partnered forms of sexual expression. We identify four determinants of sexual functioning: biologic, psychological, social context (including culture), and interactions of these with each other. Recent literature on the impact of aging and physical health documents some decline in frequency of sexual activity. Interest continues in the role of hormones in male and female sexual functioning. Recent research highlights the role of the social context, especially the presence of a sexual partner and the relationship with that partner, in sexual activity. We discuss variations in sexual functioning by life course events, gender, and race and ethnicity. Relevant results from the Global Study of Sexual Attitudes and Behaviors; Male Attitudes Regarding Sexual Health Survey; and the National Social Life, Health and Aging Project are also reviewed. PMID- 19187703 TI - Some new food for thought: the role of vitamin D in the mental health of older adults. AB - Vitamin D, a multipurpose steroid hormone vital to health, has been increasingly implicated in the pathology of cognition and mental illness. Hypovitaminosis D is prevalent among older adults, and several studies suggest an association between hypovitaminosis D and basic and executive cognitive functions, depression, bipolar disorder, and schizophrenia. Vitamin D activates receptors on neurons in regions implicated in the regulation of behavior, stimulates neurotrophin release, and protects the brain by buffering antioxidant and anti-inflammatory defenses against vascular injury and improving metabolic and cardiovascular function. Although additional studies are needed to examine the impact of supplementation on cognition and mood disorders, given the known health benefits of vitamin D, we recommend greater supplementation in older adults. PMID- 19187704 TI - Current treatments for sleep disturbances in individuals with dementia. AB - Sleep disturbances are widespread among older adults. Degenerative neurologic disorders that cause dementia, such as Alzheimer's disease and Parkinson's disease, exacerbate age-related changes in sleep, as do many common comorbid medical and psychiatric conditions. Medications used to treat chronic illness and insomnia have many side effects that can further disrupt sleep and place patients at risk for injury. This article reviews the neurophysiology of sleep in normal aging and sleep changes associated with common dementia subtypes and comorbid conditions. Current pharmacologic and nonpharmacologic evidence-based treatment options are discussed, including the use of light therapy, increased physical and social activity, and multicomponent cognitive-behavioral interventions for improving sleep in institutionalized and community-dwelling adults with dementia. PMID- 19187705 TI - Group therapy in older adults. AB - Group therapy is a well-established therapeutic modality for older individuals often conducted in combination with individual psychotherapy and pharmacotherapy. Group leaders must be aware of age-specific factors when working with older cohorts, including the presence of cognitive impairment, physical disability, and loss of independence. A limited number of outcome studies have found that various forms of group therapy in late life are generally efficacious for depression and other symptoms and compare favorably with individual psychotherapy. PMID- 19187706 TI - Metabolic risks in older adults receiving second-generation antipsychotic medication. AB - Metabolic syndrome is prevalent in older adults and increases the risk of cardiovascular disease. Second-generation antipsychotics (aripiprazole, clozapine, olanzapine, quetiapine, risperidone, and ziprasidone) increase the risk of metabolic syndrome and present many challenges for psychiatrists. In this article, we review the relationships between second-generation antipsychotics and metabolic syndrome with a focus on older adults. Because few studies focus exclusively on older adults, we augment this review with relevant findings from younger adults. The differential risk factors of each medication are reviewed, as are recent findings in monitoring and treating metabolic syndrome. Olanzapine and clozapine are more strongly associated with metabolic risks, whereas aripiprazole and ziprasidone are less associated. Although lifestyle modifications can help to reduce some aspects of metabolic syndrome, lifestyle modifications in conjunction with metformin therapy appear to be most effective. PMID- 19187708 TI - Risk factors for suicide and suicide attempts in borderline personality disorder. PMID- 19187709 TI - Prospective risk factors for suicide attempts in borderline personality disorder. PMID- 19187707 TI - Psychological distress and risk for dementia. AB - The concept of mild cognitive impairment (MCI) primarily emphasizes changes in individuals' mental abilities, but it has recently been suggested that neuropsychiatric symptoms should also be considered important factors in age related neurodegeneration. Psychological distress, defined as a reaction of an individual to external and internal stresses, is characterized by a mixture of psychological symptoms. It also may be considered a neuropsychiatric symptom encompassing depression, anxiety, and apathy. This paper reviews and summarizes recent evidence and relevant issues regarding the presence of psychological distress in healthy older adults and MCI patients and its relationship to risk for developing dementia. Results presented in this review show that psychological distress and depressive, anxious, and apathetic symptoms can be present in MCI and may predict progression to dementia. This article also provides suggestions for future research. PMID- 19187710 TI - Borderline personality disorder and couple dysfunctions. AB - The central characteristics of borderline personality disorder (BPD) are bound to be associated with the development and maintenance of couple dysfunction. Although seven of the nine diagnostic criteria of BPD in the DSM-IV-TR refer directly to interpersonal functioning, very few empiric studies have addressed the exact nature of the relationship between BPD and couple functioning. This article examines recent studies describing couple outcomes--union formation and durability, partner choice, relationship satisfaction, intimate violence, attachment security, and sexual functioning--associated with BPD. The relationship between couple dynamics (including partner personality characteristics) and BPD symptomatology is probably bidirectional or reciprocal. The review concludes with an exploration of diagnostic and treatment implications. PMID- 19187712 TI - Diagnosis of borderline personality disorder in China: current status and future directions. AB - This paper reviews the current status and future directions of borderline personality disorder (BPD) research in China. Although the committee of the third version of the Chinese Classification of Mental Disorders (CCMD-3) rejected BPD as a valid diagnostic category and instead adopted the term impulsive personality disorder (IPD), our literature review on personality disorders from 1979 to 2008 in China indicated that BPD was the most popular research topic among researchers and clinicians. Available empiric evidence on BPD in China provided preliminary support for the construct validity and clinical utility of BPD in clinical and nonclinical Chinese samples. Future studies in the following areas are suggested: 1) developing reliable assessment instruments for measuring BPD pathology in China, 2) comparing the construct validity and phenomenology of CCMD IPD and DSM BPD among Chinese patients, 3) examining potential cultural differences in symptom expression of BPD pathology among the Chinese, and 4) exploring indigenous and imported methods for treating BPD patients in China. PMID- 19187711 TI - Borderline personality disorder and childhood trauma: evidence for a causal relationship. AB - The debate over whether childhood trauma is a causative factor in the development of borderline personality disorder continues in the literature despite decades of research. This review examines this body of literature published from 1995 through 2007 to assess the strength of evidence for such a causal relationship. A unique conceptual approach was used, as we considered the literature in the context of Hill's classic criteria for demonstrating causation. Results of this review suggest that evidence supports the causal relationship, particularly if the relationship is considered as part of a multifactorial etiologic model. Directions for future research and clinical implications are discussed. PMID- 19187713 TI - Affective instability in borderline personality disorder: experience sampling findings. AB - Affective instability, defined as repeated, rapid, and abrupt shifts in mood, is considered the core pathology in borderline personality disorder. The temporal pattern of affective instability can be best captured with the experience sampling method-longitudinal assessment of people's affective states as they occur in real time and in their natural environment. A review of the experience sampling studies published to date for borderline personality disorder suggests the following mood variability pattern: intense negative mood, more frequent and abrupt mood changes than healthy controls and patients with major depression, and partial triggering of affect by external events. The method also has great potential to investigate the links between affective instability and other psychological and behavioral correlates of the disorder, such as suicide, lack of self-esteem, and erratic behaviors. However, the method requires systematic study to determine best data collection designs and mathematical models of mood variability. PMID- 19187714 TI - Dissociation and borderline personality disorder: an update for clinicians. AB - Dissociation occurs in about two thirds of people with borderline personality disorder (BPD) but is still not well understood by clinicians. In the past decade, however, research has used new measures of dissociation that provide some of the detail that clinicians need to understand and treat the dissociative symptoms of patients with BPD. In particular, this review examines BPD's comorbidity with the dissociative disorders, the neurobiology of dissociation in BPD, the role of trauma and disorganized attachment in the etiology of dissociation in BPD, and the clinical assessment and treatment of dissociation in BPD. PMID- 19187715 TI - Avoidant personality disorder and its relationship to social phobia. AB - This review summarizes past and recent findings in the empiric literature and the evolution of the concepts of avoidant personality disorder (APD) and social phobia (SP). APD is an internally consistent dimensional personality pathology that causes dysfunction that appears to be dimensional rather than a sudden jump in impairment after a certain number of criteria have been met. It has state and trait personality components. Evidence indicates that symptoms are at least partially treatable with psychological or pharmacologic interventions. APD and SP have similar symptoms and treatment response and identical genetics. We can conclude from the empiric evidence that no dividing line exists between APD and SP, with APD merely being the more severe form of the disorder. The best conceptualization is that APD is a dimensional personality pathology that in its attenuated form (SP) resembles an anxiety disorder. PMID- 19187716 TI - Fat, muscle, bone, and television. PMID- 19187717 TI - Maternal eating, infant feeding, and growth. PMID- 19187719 TI - Heart murmurs in neonates. PMID- 19187720 TI - Blood cytokines and BPD. PMID- 19187721 TI - Overall health in youth with type 1 diabetes. PMID- 19187722 TI - Diet components and insulin resistance. PMID- 19187723 TI - Birth weight and subsequent fitness. PMID- 19187724 TI - Epidemiology of seizures in term infants. PMID- 19187725 TI - Early rapid diagnostic viral tests in the emergency department setting. PMID- 19187726 TI - The fetal kidney. PMID- 19187727 TI - An initial step toward improving outcomes for preterm infants. PMID- 19187728 TI - Federation of Pediatric Organizations Task Force on Women in Pediatrics: considerations for part-time training and employment for research-intensive fellows and faculty. PMID- 19187729 TI - RBP4: from retinol transporter to biomarker? PMID- 19187730 TI - They're still kids. PMID- 19187733 TI - Ex utero intrapartum treatment procedure: a peripartum management strategy in particularly challenging cases. PMID- 19187736 TI - Use of enzyme replacement therapy (Laronidase) before hematopoietic stem cell transplantation for mucopolysaccharidosis I: experience in 18 patients. AB - We describe the use of enzyme replacement therapy in conjunction with hematopoietic stem cell transplantation in 18 consecutive patients with severe mucopolysaccharidosis type I. The survival and engraftment rate was 89% overall and 93% for the 15 patients who received full-intensity conditioning. PMID- 19187735 TI - Significant vitamin D deficiency in youth with type 1 diabetes mellitus. AB - Serum 25-hydroxyvitamin D was measured in 128 youth with type 1 diabetes mellitus. Less than 25% of the patients were vitamin D sufficient. Because individuals with type 1 diabetes mellitus possess multiple risk factors for skeletal fragility, ensuring vitamin D sufficiency throughout childhood and adolescence in this population seems especially warranted. PMID- 19187739 TI - Clinical utility of array comparative genomic hybridization: uncovering tumor susceptibility in individuals with developmental delay. AB - Microarray-based comparative genomic hybridization can determine genome-wide copy number alterations at the kilobase level. We highlight the clinical utility of microarray-based comparative genomic hybridization in determining tumor susceptibility in 3 patients with dysmorphic features and developmental delay, likely decreasing both morbidity and mortality in these patients. PMID- 19187738 TI - Congenital T cell deficiency in a patient with CHARGE syndrome. AB - CHARGE syndrome is an autosomal dominant condition caused by mutations in chromodomain helicase DNA-binding 7. We report a patient with molecularly confirmed CHARGE syndrome, which included a congenital T cell deficiency, who was treated with peripheral blood mononuclear cell transplantation. PMID- 19187741 TI - Unclear swelling of the popliteal fossa due to a giant pseudoaneurysm associated with osteochondroma. PMID- 19187742 TI - Cold-induced anaphylaxis. PMID- 19187743 TI - Nasal steroids helpful for short-term treatment of children with obstructive sleep apnea. PMID- 19187745 TI - Risk of death is not increased in children with simple febrile seizures. PMID- 19187746 TI - Regular use of inhaled corticosteroids controls symptoms of mild persistent asthma, but with growth effect. PMID- 19187747 TI - Magnesium treatment of mothers may decrease the incidence of cerebral palsy in at risk infants. PMID- 19187748 TI - Physical therapy is effective for deformational plagiocephaly. PMID- 19187749 TI - Escalating dose indomethacin for prophylactic closure of patent ductus arteriosus does not improve closure rates and is associated with increased complications. PMID- 19187750 TI - Cesarean section and risk of asthma. The role of intrapartum antibiotics: a missing piece? PMID- 19187751 TI - A case of chronic recurrent multifocal osteomyelitis successfully treated with neridronate. PMID- 19187753 TI - BRCTing up is hard to do. AB - In this issue of Molecular Cell, Usui et al. (2009) show that Mec1-phosphorylated Rad9 SCD interacts with the BRCT domain, promoting oligomerization. This sustains the DNA damage checkpoint and is suppressed by Rad53 phosphorylation in a negative feedback circuit. PMID- 19187754 TI - Assembly of export-competent mRNP: it's all about being connected. AB - In this issue of Molecular Cell, Johnson et al. (2009) reveal a physical link between the yeast Pcf11p and Yra1p proteins, providing insights into the coupling of mRNA 3' end formation and export. PMID- 19187755 TI - Less is more for leaderless mRNA translation. AB - In this issue of Molecular Cell, Kaberdina et al. (2009) show that prolonged exposure of ribosomes to the antibiotic kasugamycin triggers the spontaneous loss of small subunit proteins and produces a reduced ribosomal particle that exclusively translates leaderless mRNAs. PMID- 19187756 TI - TgI4 lipase: a big fat target for cell-cycle entry. AB - In a recent issue of Molecular Cell, Kurat et al. (2009) find that when quiescent yeast re-enter the cell cycle, the cell-cycle cyclin-dependent kinase phosphorylates and activates the lipase TgI4, liquidating fat and providing fatty acids for cell-cycle entry. PMID- 19187757 TI - Deadly splicing: Bax becomes Almighty. AB - Activation of Bax and Bak by BH3-only molecules triggers mitochondrial apoptosis. In a recent issue of Molecular Cell, Fu et al. (2009) identify a constitutively active isoform of Bax, Baxbeta, whose activity is tightly controlled by the ubiquitin-proteasome system. PMID- 19187758 TI - Maintenance of the DNA-damage checkpoint requires DNA-damage-induced mediator protein oligomerization. AB - Oligomeric assembly of Brca1 C-terminal (BRCT) domain-containing mediator proteins occurs at sites of DNA damage. However, the functional significance and regulation of such assemblies are not well understood. In this study, we defined the molecular mechanism of DNA-damage-induced oligomerization of the S. cerevisiae BRCT protein Rad9. Our data suggest that Rad9's tandem BRCT domain mediates Rad9 oligomerization via its interaction with its own Mec1/Tel1 phosphorylated SQ/TQ cluster domain (SCD). Rad53 activation is unaffected by mutations that impair Rad9 oligomerization, but checkpoint maintenance is lost, indicating that oligomerization is required to sustain checkpoint signaling. Once activated, Rad53 phosphorylates the Rad9 BRCT domain, which attenuates the BRCT SCD interaction. Failure to phosphorylate the Rad9 BRCT results in cytologically visible Rad9 foci. This suggests a feedback loop wherein Rad53 activity and Rad9 oligomerization are regulated to tune the DNA-damage response. PMID- 19187760 TI - Actin homolog MreB affects chromosome segregation by regulating topoisomerase IV in Escherichia coli. AB - In Escherichia coli, topoisomerase IV, a type II topoisomerase, mediates the resolution of topological linkages between replicated daughter chromosomes and is essential for chromosome segregation. Topo IV activity is restricted to only a short interval late in the cell cycle. However, the mechanism that confers this temporal regulation is unknown. Here we report that the bacterial actin homolog MreB participates in the temporal oscillation of Topo IV activity. We show that mreB mutant strains are deficient in Topo IV activity. In addition, we demonstrate that, depending upon whether it is in a monomeric or polymerized state, MreB affects Topo IV activity differentially. In addition, MreB physically interacts with the ParC subunit of Topo IV. Together, these results may explain how dynamics of the bacterial cytoskeleton are coordinated with the timing of chromosome segregation. PMID- 19187759 TI - Second-end capture in DNA double-strand break repair promoted by Brh2 protein of Ustilago maydis. AB - Brh2 plays a central role in the homologous recombination system of Ustilago maydis, mediating delivery of Rad51 to single-stranded DNA. Here we report that Brh2 can pair the displaced strand of a D loop with a complementary single stranded DNA to form a duplexed, or double, D loop. The reaction emulates the second-end capture step envisioned in models of DNA double-strand break repair. This second-end capture reaction promoted by Brh2 proceeds efficiently when performed in the presence of Rad51 under conditions that block annealing by Rad52, or when the second single-stranded DNA substrate is replaced by double stranded DNA. In a coupled reaction that requires extension of the D loop more than 200 nt by DNA synthesis in order to reveal a complementary region, Brh2 was also able to promote second-end capture and thus model a synthesis-dependent strand-annealing mechanism. PMID- 19187761 TI - Structural basis for the requirement of additional factors for MLL1 SET domain activity and recognition of epigenetic marks. AB - The mixed-lineage leukemia protein MLL1 is a transcriptional regulator with an essential role in early development and hematopoiesis. The biological function of MLL1 is mediated by the histone H3K4 methyltransferase activity of the carboxyl terminal SET domain. We have determined the crystal structure of the MLL1 SET domain in complex with cofactor product AdoHcy and a histone H3 peptide. This structure indicates that, in order to form a well-ordered active site, a highly variable but essential component of the SET domain must be repositioned. To test this idea, we compared the effect of the addition of MLL complex members on methyltransferase activity and show that both RbBP5 and Ash2L but not Wdr5 stimulate activity. Additionally, we have determined the effect of posttranslational modifications on histone H3 residues downstream and upstream from the target lysine and provide a structural explanation for why H3T3 phosphorylation and H3K9 acetylation regulate activity. PMID- 19187762 TI - Enhancement of the seed-target recognition step in RNA silencing by a PIWI/MID domain protein. AB - Target recognition in RNA silencing is governed by the "seed sequence" of a guide RNA strand associated with the PIWI/MID domain of an Argonaute protein in RISC. Using a reconstituted in vitro target recognition system, we show that a model PIWI/MID domain protein confers position-dependent tightening and loosening of guide-strand-target interactions. Over the seed sequence, the interaction affinity is enhanced up to approximately 300-fold. Enhancement is achieved through a reduced entropy penalty for the interaction. In contrast, interactions 3' of the seed are inhibited. We quantified mismatched target recognition inside and outside the seed, revealing amplified discrimination at the third position in the seed mediated by the PIWI/MID domain. Thus, association of the guide strand with the PIWI/MID domain generates an enhanced affinity anchor site over the seed that can promote fidelity in target recognition and stabilize and guide the assembly of the active silencing complex. PMID- 19187763 TI - An unexpected type of ribosomes induced by kasugamycin: a look into ancestral times of protein synthesis? AB - Translation of leaderless mRNAs, lacking ribosomal recruitment signals other than the 5'-terminal AUG-initiating codon, occurs in all three domains of life. Contemporary leaderless mRNAs may therefore be viewed as molecular fossils resembling ancestral mRNAs. Here, we analyzed the phenomenon of sustained translation of a leaderless mRNA in the presence of the antibiotic kasugamycin. Unexpected from the known in vitro effects of the drug, kasugamycin induced the formation of stable approximately 61S ribosomes in vivo, which were proficient in selectively translating leaderless mRNA. 61S particles are devoid of more than six proteins of the small subunit, including the functionally important proteins S1 and S12. The lack of these proteins could be reconciled with structural changes in the 16S rRNA. These studies provide in vivo evidence for the functionality of ribosomes devoid of multiple proteins and shed light on the evolutionary history of ribosomes. PMID- 19187765 TI - ING4 mediates crosstalk between histone H3 K4 trimethylation and H3 acetylation to attenuate cellular transformation. AB - Aberrations in chromatin dynamics play a fundamental role in tumorigenesis, yet relatively little is known of the molecular mechanisms linking histone lysine methylation to neoplastic disease. ING4 (Inhibitor of Growth 4) is a native subunit of an HBO1 histone acetyltransferase (HAT) complex and a tumor suppressor protein. Here we show a critical role for specific recognition of histone H3 trimethylated at lysine 4 (H3K4me3) by the ING4 PHD finger in mediating ING4 gene expression and tumor suppressor functions. The interaction between ING4 and H3K4me3 augments HBO1 acetylation activity on H3 tails and drives H3 acetylation at ING4 target promoters. Further, ING4 facilitates apoptosis in response to genotoxic stress and inhibits anchorage-independent cell growth, and these functions depend on ING4 interactions with H3K4me3. Together, our results demonstrate a mechanism for brokering crosstalk between H3K4 methylation and H3 acetylation and reveal a molecular link between chromatin modulation and tumor suppressor mechanisms. PMID- 19187764 TI - Oncogenic B-RAF negatively regulates the tumor suppressor LKB1 to promote melanoma cell proliferation. AB - The LKB1-AMPK signaling pathway serves as a critical cellular sensor coupling energy homeostasis to cell growth, proliferation, and survival. However, how tumor cells suppress this signaling pathway to gain growth advantage under conditions of energy stress is largely unknown. Here, we show that AMPK activation is suppressed in melanoma cells with the B-RAF V600E mutation and that downregulation of B-RAF signaling activates AMPK. We find that in these cells LKB1 is phosphorylated by ERK and Rsk, two kinases downstream of B-RAF, and that this phosphorylation compromises the ability of LKB1 to bind and activate AMPK. Furthermore, expression of a phosphorylation-deficient mutant of LKB1 allows activation of AMPK and inhibits melanoma cell proliferation and anchorage independent cell growth. Our findings provide a molecular linkage between the LKB1-AMPK and the RAF-MEK-ERK pathways and suggest that suppression of LKB1 function by B-RAF V600E plays an important role in B-RAF V600E-driven tumorigenesis. PMID- 19187766 TI - HBO1 HAT complexes target chromatin throughout gene coding regions via multiple PHD finger interactions with histone H3 tail. AB - The HBO1 HAT protein is the major source of histone H4 acetylation in vivo and has been shown to play critical roles in gene regulation and DNA replication. A distinctive characteristic of HBO1 HAT complexes is the presence of three PHD finger domains in two different subunits: tumor suppressor proteins ING4/5 and JADE1/2/3. Biochemical and functional analyses indicate that these domains interact with histone H3 N-terminal tail region, but with a different specificity toward its methylation status. Their combinatorial action is essential in regulating chromatin binding and substrate specificity of HBO1 complexes, as well as cell growth. Importantly, localization analyses on the human genome indicate that HBO1 complexes are enriched throughout the coding regions of genes, supporting a role in transcription elongation. These results underline the importance and versatility of PHD finger domains in regulating chromatin association and histone modification crosstalk within a single protein complex. PMID- 19187768 TI - Why we should put clothes on mice. AB - Mitochondrial uncoupling protein 1 (UCP1) is a key regulator of adaptive thermogenesis and energy expenditure. Mice lacking UCP1 are cold sensitive, but surprisingly not obese at room temperature. In this issue of Cell Metabolism, Feldmann et al. (2009) unmask an obesogenic phenotype by simply maintaining these mice at thermoneutrality. PMID- 19187767 TI - Symmetric signaling by an asymmetric 1 erythropoietin: 2 erythropoietin receptor complex. AB - Via sites 1 and 2, erythropoietin binds asymmetrically to two identical receptor monomers, although it is unclear how asymmetry affects receptor activation and signaling. Here we report the design and validation of two mutant erythropoietin receptors that probe the role of individual members of the receptor dimer by selectively binding either site 1 or site 2 on erythropoietin. Ba/F3 cells expressing either mutant receptor do not respond to erythropoietin, but cells co expressing both receptors respond to erythropoietin by proliferation and activation of the JAK2-Stat5 pathway. A truncated receptor with only one cytosolic tyrosine (Y343) is sufficient for signaling in response to erythropoietin, regardless of the monomer on which it is located. Similarly, only one receptor in the dimer needs a juxtamembrane hydrophobic L253 or W258 residue, essential for JAK2 activation. We conclude that despite asymmetry in the ligand receptor interaction, both sides are competent for signaling, and appear to signal equally. PMID- 19187769 TI - Fat rationing in dauer times. AB - The fundamental task of maintaining energy balance is complex when nutrient levels are plentiful, but it becomes even more challenging when nutrients are dynamic or scarce. A recent Nature report delineates a role of the AMP kinase pathway in rationing energy stores for the long-term survival of Caenorhabditis elegans dauers (Narbonne and Roy, 2009). PMID- 19187771 TI - CXCL10 impairs beta cell function and viability in diabetes through TLR4 signaling. AB - In type 1 and type 2 diabetes (T1/T2DM), beta cell destruction by apoptosis results in decreased beta cell mass and progression of the disease. In this study, we found that the interferon gamma-inducible protein 10 plays an important role in triggering beta cell destruction. Islets isolated from patients with T2DM secreted CXCL10 and contained 33.5-fold more CXCL10 mRNA than islets from control patients. Pancreatic sections from obese nondiabetic individuals and patients with T2DM and T1DM expressed CXCL10 in beta cells. Treatment of human islets with CXCL10 decreased beta cell viability, impaired insulin secretion, and decreased insulin mRNA. CXCL10 induced sustained activation of Akt, JNK, and cleavage of p21-activated protein kinase 2 (PAK-2), switching Akt signals from proliferation to apoptosis. These effects were not mediated by the commonly known CXCL10 receptor CXCR3 but through TLR4. Our data suggest CXCL10 as a binding partner for TLR4 and as a signal toward beta cell failure in diabetes. PMID- 19187770 TI - The geometry of leptin action in the brain: more complicated than a simple ARC. AB - Leptin signals the repletion of fat stores, acting in the CNS to permit energy utilization by a host of autonomic and neuroendocrine processes and to decrease feeding. While much recent research has focused on the leptin-regulated circuitry of the hypothalamic arcuate nucleus (ARC), the majority of brain leptin receptor (LepRb)-expressing neurons lie outside the ARC in other CNS regions known to modulate energy balance. Each set of LepRb neurons throughout the brain presumably mediates unique aspects of leptin action, and understanding the function for LepRb-expressing neurons throughout the brain represents a crucial next step in the study of energy homeostasis. PMID- 19187772 TI - Secondary consequences of beta cell inexcitability: identification and prevention in a murine model of K(ATP)-induced neonatal diabetes mellitus. AB - ATP-insensitive K(ATP) channel mutations cause neonatal diabetes mellitus (NDM). To explore the mechanistic etiology, we generated transgenic mice carrying an ATP insensitive mutant K(ATP) channel subunit. Constitutive expression in pancreatic beta cells caused neonatal hyperglycemia and progression to severe diabetes and growth retardation, with loss of islet insulin content and beta cell architecture. Tamoxifen-induced expression in adult beta cells led to diabetes within 2 weeks, with similar secondary consequences. Diabetes was prevented by transplantation of normal islets under the kidney capsule. Moreover, the endogenous islets maintained normal insulin content and secretion in response to sulfonylureas, but not glucose, consistent with reduced ATP sensitivity of beta cell K(ATP) channels. In NDM, transfer to sulfonylurea therapy is less effective in older patients. This may stem from poor glycemic control or lack of insulin because glibenclamide treatment prior to tamoxifen induction prevented diabetes and secondary complications in mice but failed to halt disease progression after diabetes had developed. PMID- 19187774 TI - Cideb, an ER- and lipid droplet-associated protein, mediates VLDL lipidation and maturation by interacting with apolipoprotein B. AB - Secretion of triacylglycerol-enriched very-low-density lipoproteins (VLDLs) from the liver is vital for maintaining plasma lipid homeostasis. However, the process of VLDL assembly and lipidation is not well characterized. Here, we observed that liver of Cideb null mice had higher levels of triacylglycerols accompanied by low level of VLDL secretion. Furthermore, VLDL particles secreted from hepatocytes of Cideb null mice have low levels of triacylglycerols but normal levels of apoB. We also observed that Cideb is localized to endoplasmic reticulum and lipid droplets. Importantly, we have identified apoB as a Cideb-interacting protein. By infecting adenoviruses expressing various Cideb truncations into hepatocytes of Cideb null mice, we found that Cideb requires both its apoB-binding and lipid droplet association domains to restore the secretion of triacylglycerol-enriched VLDL particles. Our data suggest that Cideb promotes the formation of triacylglycerol-enriched VLDL particles and provides a molecular insight into VLDL lipidation and maturation in hepatocytes. PMID- 19187773 TI - Molecular mechanisms of hepatic steatosis and insulin resistance in the AGPAT2 deficient mouse model of congenital generalized lipodystrophy. AB - Mutations in 1-acylglycerol-3-phosphate-O-acyltransferase 2 (AGPAT2) cause congenital generalized lipodystrophy. To understand the molecular mechanisms underlying the metabolic complications associated with AGPAT2 deficiency, Agpat2 null mice were generated. Agpat2(-/-) mice develop severe lipodystrophy affecting both white and brown adipose tissue, extreme insulin resistance, diabetes, and hepatic steatosis. The expression of lipogenic genes and rates of de novo fatty acid biosynthesis were increased approximately 4-fold in Agpat2(-/-) mouse livers. The mRNA and protein levels of monoacylglycerol acyltransferase isoform 1 were markedly increased in the livers of Agpat2(-/-) mice, suggesting that the alternative monoacylglycerol pathway for triglyceride biosynthesis is activated in the absence of AGPAT2. Feeding a fat-free diet reduced liver triglycerides by approximately 50% in Agpat2(-/-) mice. These observations suggest that both dietary fat and hepatic triglyceride biosynthesis via a monoacylglycerol pathway may contribute to hepatic steatosis in Agpat2(-/-) mice. PMID- 19187775 TI - Fasting-induced hypothermia and reduced energy production in mice lacking acetyl CoA synthetase 2. AB - Acetate is activated to acetyl-CoA by acetyl-CoA synthetase 2 (AceCS2), a mitochondrial enzyme. Here, we report that the activation of acetate by AceCS2 has a specific and unique role in thermogenesis during fasting. In the skeletal muscle of fasted AceCS2(-/-) mice, ATP levels were reduced by 50% compared to AceCS2(+/+) mice. Fasted AceCS2(-/-) mice were significantly hypothermic and had reduced exercise capacity. Furthermore, when fed a low-carbohydrate diet, 4-week old weaned AceCS2(-/-) mice also exhibited hypothermia accompanied by sustained hypoglycemia that led to a 50% mortality. Therefore, AceCS2 plays a significant role in acetate oxidation needed to generate ATP and heat. Furthermore, AceCS2(-/ ) mice exhibited increased oxygen consumption and reduced weight gain on a low carbohydrate diet. Our findings demonstrate that activation of acetate by AceCS2 plays a pivotal role in thermogenesis, especially under low-glucose or ketogenic conditions, and is crucially required for survival. PMID- 19187776 TI - UCP1 ablation induces obesity and abolishes diet-induced thermogenesis in mice exempt from thermal stress by living at thermoneutrality. AB - As original studies of UCP1-ablated mice failed to demonstrate an obesogenic effect, alternative mechanisms for adaptive adrenergic thermogenesis have been sought. However, we demonstrate here that in C57Bl6 mice exempt from thermal stress (i.e., kept at thermoneutrality), UCP1 ablation in itself induced obesity, even in mice fed control diet, and vastly augmented diet-induced obesity (high fat diet); i.e., the mice exhibited increased metabolic efficiency. In wild-type mice, high-fat diet increased norepinephrine-induced thermogenesis; i.e., diet induced thermogenesis was observed, but no such effect was observed in UCP1 ablated mice, demonstrating that diet-induced thermogenesis fully emanates from UCP1 activity. We conclude that ambient temperature is qualitatively determinative for the outcome of metabolic studies, that no other protein and no other mechanism can substitute for UCP1 in mediating diet-induced adrenergic thermogenesis, and that UCP1 activity can be determinative for obesity development in mice and possibly in humans. PMID- 19187777 TI - Structures of Leishmania major orthologues of macrophage migration inhibitory factor. AB - Leishmania major, an intracellular parasitic protozoon that infects, differentiates and replicates within macrophages, expresses two closely related MIF-like proteins. To ascertain the roles and potential differences of these two Leishmania proteins, recombinant L. major MIF1 and MIF2 have been produced and the structures resolved by X-ray crystallography. Each has a trimeric ring architecture similar to mammalian MIF, but with some structurally distinct features. LmjMIF1, but not LmjMIF2, has tautomerase activity. LmjMIF2 is found in all life cycle stages whereas LmjMIF1 is found exclusively in amastigotes, the intracellular stage responsible for mammalian disease. The findings are consistent with parasite MIFs modulating or circumventing the host macrophage response, thereby promoting parasite survival, but suggest the LmjMIFs have potentially different biological roles. Analysis of the Leishmania braziliensis genome showed that this species lacks both MIF genes. Thus MIF is not a virulence factor in all species of Leishmania. PMID- 19187778 TI - Cryptosporidium: genomic and biochemical features. AB - Recent progress in understanding the unique biochemistry of the two closely related human enteric pathogens Cryptosporidium parvum and Cryptosporidium hominis has been stimulated by the elucidation of the complete genome sequences for both pathogens. Much of the work that has occurred since that time has been focused on understanding the metabolic pathways encoded by the genome in hopes of providing increased understanding of the parasite biology, and in the identification of novel targets for pharmacological interventions. However, despite identifying the genes encoding enzymes that participate in many of the major metabolic pathways, only a hand full of proteins have actually been the subjects of detailed scrutiny. Thus, much of the biochemistry of these parasites remains a true mystery. PMID- 19187779 TI - Essential role of SBP-1 activation in oxygen deprivation induced lipid accumulation and increase in body width/length ratio in Caenorhabditis elegans. AB - Epidemiological evidence suggests a link between chronic oxygen starvation and fat accumulation/obesity, however the underlying mechanism remains unclear. Using Caenorhabditis elegans we found extended oxygen deprivation resulted in activation of SBP-1, the worm homologue of SREBP1, a transcription factor important in maintaining lipid homeostasis. SBP-1 knockdown prevented hypoxia induced fat accumulation and the associated increase in worm width/length ratio, demonstrating that SBP-1/SREBP1 plays an essential role in hypoxia-induced lipid accumulation and body shape alteration. This study provides the first evidence suggesting that activation of SREBP1 may be a critical pathogenic factor contributing to chronic hypoxia associated excessive fat accumulation/obesity in humans. PMID- 19187780 TI - Prominent expression of FRS2beta protein in neural cells and its association with intracellular vesicles. AB - The fibroblast growth factor receptor substrate (FRS)-2 protein family comprises FRS2alpha, a well-known central mediator for fibroblast growth factor signaling, and FRS2beta, whose endogenous expression pattern and function are not yet defined. Immunohistochemical analysis revealed that expression of FRS2beta was restricted to neural tissues and it colocalized with Tuj1, a neuronal marker. There are two distinct patterns of FRS2beta expression in neural cells: punctate and cup/ring-shaped; moreover, some particles colocalized with lysosomes. Stimulation with brain-derived neurotrophic factor (BDNF) enhanced FRS2beta phosphorylation and the cup/ring-shaped pattern. These results suggest a probable role of FRS2beta in the intracellular degradation systems of neural cells, which involves lysosomes. PMID- 19187781 TI - Narrow-band imaging without optical magnification for histologic analysis of colorectal polyps. AB - BACKGROUND & AIMS: The ability to determine colorectal polyp pathology by endoscopy could reduce the risks of polypectomy and the cost of pathologic evaluation. This study evaluated the ability of the Olympus Exera 180 high definition colonoscope (Olympus America, Inc, Center Valley, PA), with narrow band imaging, to predict colorectal polyp histology. METHODS: A library of 320 endoscopic photographs with correlated histologic information was used to identify endoscopic features associated with adenomatous and hyperplastic histology. These features were tested in a prospective study of 451 consecutively identified colorectal polyps. Polyps were observed endoscopically and assigned a designation of high or low confidence. The primary end points were the predictive value of high-confidence endoscopic interpretations of adenoma and hyperplastic histology for polyps 5 mm and smaller in size. RESULTS: Endoscopic predictions of adenoma and hyperplastic histology were made with high confidence for 80% and 83% of cases, respectively. High-confidence predictions were more likely than low confidence predictions to be correct (P<.001). High-confidence predictions of adenoma and hyperplastic histology were correct for 91% and 95%, respectively, of polyps 5 mm and smaller in size. CONCLUSIONS: Introduction of confidence levels to the endoscopic interpretation of colorectal polyp histology allows sufficient accuracy for the use of the Exera narrow-band imaging system in the identification of distal hyperplastic polyps that do not need resection, as well as to plan postpolypectomy surveillance without pathologic evaluation of polyps 5 mm in size or smaller. PMID- 19187783 TI - A rapid and efficient strategy to generate allele-specific anti-HLA monoclonal antibodies. AB - That generation of allele-specific anti-human leukocyte antigen (HLA) monoclonal antibodies (ASHmAb) is very difficult is well known. This is thought to be due to the unique epitope structure, an assemblage of amino acid residues that lie separately in the amino acid sequence of human HLA, and to its low antigenicity compared with that of common epitopes recognized as xenogeneic determinants by mice. Here we report a rapid and efficient strategy to generate ASHmAb. Different from usual immunization methods is that we suppressed the production of non allele-specific anti-HLA antibodies against xenogeneic determinants of HLA molecules by immunizing human HLA-B51 transgenic mice against non-HLA-B51 HLA tetramers. In addition, HLA-coated beads enabled rapid and efficient screening for ASHmAb. ASHmAb generated by this strategy will be useful for HLA typing and for clinical diagnosis, such as flow cytometry-based chimerism analysis for early detection of graft failure and relapse of leukemia after HLA-mismatched hematopoietic stem cell transplantation. PMID- 19187782 TI - Engineered cell surface expression of membrane immunoglobulin as a means to identify monoclonal antibody-secreting hybridomas. AB - Monoclonal antibodies (mAbs) have proven to be effective biological reagents in the form of therapeutic drugs and diagnostics for many pathologies, as well as valuable research tools. Existing methods for isolating mAb-producing hybridomas are tedious and time consuming. Herein we describe a novel system in which mAb secreting hybridoma cells were induced to co-express significant amounts of the membrane form of the secreted immunoglobulin (Ig) on their surfaces and are efficiently recovered by fluorescent activated cell sorting (FACS). Fusion of a novel myeloma parent, SP2ab, expressing transgenic Igalpha and Igbeta of the B cell receptor complex (BCR) with spleen cells resulted in hybridomas demonstrating order of magnitude increases in BCR surface expression. Surface Ig levels correlated with transgenic Igalpha expression, and these cells also secreted normal levels of mAb. Hundreds of hybridoma lines producing mAbs specific for a variety of antigens were rapidly isolated as single cell-derived clones after FACS. Significant improvements using the Direct Selection of Hybridomas (DiSH) by FACS include reduced time and labor, improved capability of isolating positive hybridomas, and the ease of manipulating cloned cell lines relative to previously existing approaches that require Limiting Dilution Subcloning (LDS). PMID- 19187784 TI - Hypoxia influences CD40-CD40L mediated inflammation in endothelial and monocytic cells. AB - The interaction between CD40 and its ligand (CD40L) has been implicated in the pathogenesis of atherosclerosis and is recognized as a central event in the development of immuno-inflammatory processes. Our previous studies have shown that the CD40-CD40L interaction modulates platelet, neutrophil, and endothelial reactive oxygen species (ROS) generation. Hypoxia, known to be associated with tissue ischemia and inflammation, also influences the ROS production and changes the cellular redox state. However, the effect of hypoxia on CD40-CD40L mediated vascular inflammation is unknown. We have investigated whether hypoxia influences CD40-CD40L mediated vascular inflammatory responses, ROS production, and cellular interactions. We found that hypoxia significantly enhances the inflammatory effect of CD40L in both endothelial and monocytic cells (THP1). CD40-CD40L interaction in the presence of hypoxia induces ROS production, the synthesis of an inflammatory adhesive protein intercellular adhesion molecule 1 (ICAM1) and activates stress response proteins (p38 MAP kinase and HSP27), indicating that CD40L mediates the induction of oxidative stress in these cells. Importantly, we found that the effects of CD40L can be transmitted between HUVECs and monocytic THP1 cells through intercellular CD40-CD40L interaction and these processes are augmented under hypoxia. Together, these data indicate that under hypoxic conditions the CD40-CD40L interaction significantly influences adhesion molecule expression, stress generation, actin polymerization, and monocytic adhesion to endothelial cells in addition to changes in signaling. In summary, we show that hypoxia can alter CD40-CD40L mediated endothelial-monocyte interaction, playing a significant role in vascular inflammation and cellular adhesion processes. PMID- 19187785 TI - Rapid diagnosis of turbot reddish body iridovirus in turbot using the loop mediated isothermal amplification method. AB - Turbot reddish body iridovirus (TRBIV) is a new piscine iridovirus that infects the turbot, Scophthalmus maximus, cultured in northern China and can cause high mortality. In this study, a loop-mediated isothermal amplification (LAMP) method was developed for the specific detection of this virus using primers designed from an Msp I restriction DNA fragment of the TRBIV genome. Mg(2+) concentrations, the reaction temperature, and the reaction time of LAMP were optimized to 6mM, 65 degrees C, and 60 min, respectively. The detection limit of the LAMP method was as low as seven copies and was 100 times more sensitive than the conventional PCR technique. Visual inspection of LAMP amplifications demonstrated that the positive and negative reactions exhibit distinct and different colors in daylight, which means that gel electrophoresis is not necessary to judge the presence or absence of the virus. LAMP can be conducted in 1h and requires only a simple heating device for incubation. Thus, the LAMP-TRBIV detection protocol has great potential for use in the detection of TRBIV in both the laboratory and the farm. PMID- 19187786 TI - Disruption of HPV 16 E1 and E2 genes in precancerous cervical lesions. AB - The presence of HPV 16 E1 and E2 genes was detected in cervical cytological samples using polymerase chain reaction assays. A total of 48 samples were analyzed from patients with HPV 16 infections associated with 13 low-grade cervical intraepithelial neoplasia and 35 high-grade cervical intraepithelial neoplasia. Disruption/deletion sites, within E1 and E2 genes, were detected using 6 primer pairs spanning the entire gene sequences. This technique is not able to recognize mixed DNA forms (integrated plus episomal DNA); therefore, it detects only the presence of pure integrated DNA. Both E1 and E2 genes were detected in 84.6% and in 62.9% of low and high-grade lesions, respectively. The rate of samples with disrupted/deleted genes was significantly higher in high-grade cervical intraepithelial neoplasia than in low-grade cervical intraepithelial neoplasia (P<0.05). In high-grade cervical intraepithelial neoplasia the disruption/deletion pattern involved both E1 and E2 genes and E2 gene was always involved, while in the low grade cervical intraepithelial neoplasia only E1 gene was involved. In conclusion, in high-grade cervical lesions E2 gene seems a suitable target to identify HPV 16 DNA integration into cellular genome. PMID- 19187787 TI - Rapid and sensitive detection of hepatitis B virus 1762T/1764A double mutation from hepatocellular carcinomas using LNA-mediated PCR clamping and hybridization probes. AB - The 1762T/1764A double mutation of the hepatitis B virus (HBV) basal core promoter has been suggested to be a potential biomarker for hepatocellular carcinoma (HCC) among individuals with chronic HBV infection. In this study, a real-time PCR assay is established using the hybridization probes and an oligonucleotide clamp containing locked nucleic acids (LNAs). The LNA-containing oligonucleotide clamp specific for the wild type HBV is able to suppress the amplification of the wild type HBV templates. In addition, the clamp can inhibit the binding of the WT templates to the fluorescence probes thereby suppress the wild type HBV signals during the melting curve analyses. These effects facilitated the detection of HBV double mutation in the presence of 3000-fold excess of the wild type genome. Thus PCR amplification coupled with the melting curve analyses provides a quick, simple, and highly sensitive tool for the detection of this HBV double mutation. PMID- 19187788 TI - Cloning of Gallid herpesvirus 3 (Marek's disease virus serotype-2) genome as infectious bacterial artificial chromosomes for analysis of viral gene functions. AB - Marek's disease virus serotype 2 (Gallid herpesvirus 3) is a non-pathogenic alphaherpesvirus belonging to the Mardivirus genus, used widely in live vaccines against Marek's disease. Although the complete genome sequence of the MDV-2 strain HPRS-24 has been published, very little is known about the gene functions. As a first step for carrying out functional genomic analysis of MDV-2, the full length genome of the MDV-2 vaccine strain SB-1 was cloned as an infectious bacterial artificial chromosome (BAC) clone pSB-1. Virus reconstituted from the pSB-1 clone showed morphological and growth characteristics in cell culture very similar to the parent virus. Generation of SB-1 constructs deleted in glycoprotein E and viruses expressing Citrine-UL35 fusion protein by the application of different BAC mutagenesis techniques demonstrated the amenability of the pSB-1 clone for reverse genetics approaches to identify molecular determinants associated with different biological features of this virus. The generation of replication-competent infectious clones of SB-1, together with those of CVI988 and herpesvirus of turkey strains described previously, completes the portfolio of generating infectious BAC clones of the MD vaccine strains belonging to all the three serotypes, paving the way for the application of reverse genetics for functional analysis of immunogenic determinants of these vaccines as well as for developing novel recombinant vectors. PMID- 19187789 TI - Centrifugation improves the detection of HIV-1 p24 antigen in plasma from children born to mothers infected with HIV-1. AB - Detection of HIV proteins and/or nucleic acids is necessary for the diagnosis of perinatal HIV infection. Despite its low sensitivity, detection of p24 antigen in plasma is a simple and economic method for the diagnosis of HIV in exposed children. The aim of this study was to improve the sensitivity of detection of p24 using centrifugation of plasma. Forty-seven selected stored samples from 37 children (23 infected, 14 uninfected, median age of 137 days) were examined. Plasma samples (volume 0.3-1.5 ml) were defrosted, centrifuged at 23,500 x g at 4 degrees C for 60 min and determination of p24 was carried out in the resuspended pellet (0.12 ml). In 32 plasma samples from infected children, p24 was found originally in 6 (18.7%) and resulted positive in 24 (75%) pellets. When only one sample per child was considered, sensitivity was significantly higher in pellets, 3/23 uncentrifuged plasma samples and 15/23 pellets (McNemar Test, p<0.001). Specificity was 100%. The absorbance/cut-off ratio was always higher in the pellets from positive children (p=0.028). Plasma samples with volumes of 1 ml or more achieved a higher sensitivity (91.7% vs. 36.4%, p=0.009). Centrifugation of plasma samples prior to determination of p24 in pediatric patients resulted in a significant increase in sensitivity. PMID- 19187790 TI - Optimisation of a single-primer sequence-independent amplification (SP-SIA) assay: detection of previously undetectable norovirus strains associated with outbreaks of gastroenteritis. AB - The introduction of molecular diagnostic methods for investigation of gastroenteritis has significantly reduced the diagnostic gap. However, approximately 25% of cases of gastroenteritis remain undiagnosed even after screening for bacteria, parasites and viruses using the most sensitive PCR and RT PCR methods available. In recent years, it has become apparent that viruses are responsible for the majority of outbreaks of gastroenteritis. In this study, a panel of samples from outbreaks of gastroenteritis for which no aetiological agent had been identified was selected for investigation by random amplification molecular methods. An algorithm for virus purification and concentration was developed followed by a single-primer sequence-independent amplification method. These methods resulted in the identification of viruses in 5 out of 51 previously negative outbreaks. Noroviruses undetectable using two available broadly reactive diagnostic methods were detected in 4 of these outbreaks. PMID- 19187791 TI - Degenerate primers based RT-PCR for rapid detection and differentiation of airborne chicken Newcastle disease virus in chicken houses. AB - Airborne Newcastle disease (ND) viruses in the air of five chicken houses were detected and differentiated by reverse transcriptase polymerase chain reaction (RT-PCR) using degenerate primers. Fifteen air samples were collected with All Glass Impinger-30 (AGI-30) air samplers in each house. Airborne ND viruses were also isolated and virulence identified by in vivo tests. Avirulent viruses were detected both in air samples and swab samples in four houses by degenerate primers based RT-PCR. Virulent viruses were detected only in the air samples by degenerate primers based RT-PCR in two houses. Seven strains viruses were isolated from the RT-PCR positive air samples. Of the seven strains, three strains were virulent viruses and four strains were avirulent viruses identified by in vivo tests. The results showed that it was feasible to detect and differentiate NDV in the air samples using degenerate primers based RT-PCR. This technique could decrease the time it required identify NDV infected flocks while distinguishing between virulent and avirulent viruses. It will help effectively to control Newcastle disease. PMID- 19187792 TI - Antiviral activity of highly potent siRNAs against echovirus 30 and its receptor. AB - RNA interference (RNAi) has been shown to be suitable to inhibit viruses in experimental setups and is considered a promising antiviral strategy that is currently being tested in various clinical trials. The present study provides an approach to design siRNAs with high potency against a virus-specific target gene. In recent years, several outbreaks of aseptic meningitis caused by an echovirus 30 (EV-30) infection have been described. Based on an initial set of 30 in silico designed siRNAs, six siRNAs targeting the 3D RNA-dependent RNA-Polymerase (3D(Pol)) of EV-30 were selected. All but one of them showed high efficiency in both, reporter and virus assays. A second aim of the study was to re-investigate the relevance of the decay-accelerating factor (DAF, also known as CD55) as cellular entry receptor of EV-30 by means of RNAi, a question which had been under debate in previous studies. Knockdown of DAF inhibited drastically infection by EV-30 indicating that DAF plays an important role either as an attachment factor or as a receptor. PMID- 19187793 TI - GSK983: a novel compound with broad-spectrum antiviral activity. AB - GSK983, a novel tetrahydrocarbazole, inhibits the replication of a variety of unrelated viruses in vitro with EC(50) values of 5-20 nM. Both replication of the adenovirus Ad-5 and the polyoma virus SV-40, and episomal maintenance of human papillomaviruses (HPV) and Epstein-Barr virus (EBV) are susceptible to GSK983. The compound does not inhibit all viruses; herpes simplex virus (HSV-1), human immunodeficiency virus (HIV), and lytic replication of EBV were not susceptible at concentrations below 1 microM. GSK983 does inhibit the growth of cell lines immortalized by HTLV-1, EBV, HPV, SV40 and Ad-5, with EC(50) values in the range of 10-40 nM. Depending on the cell line, the compound induces either apoptosis or cytostasis at concentrations over 20 nM. GSK983 also inhibits cell lines immortalized by non-viral mechanisms, but has little effect on primary cells. The CC(50) values for keratinocytes, fibroblasts, lymphocytes, endothelial, and bone marrow progenitor cells are all above 10 microM. The pattern of inhibition, which includes diverse viruses as well as growth of immortalized cells of varied origins, suggests the target is a host cell protein, rather than a viral protein. Preliminary mechanism studies indicate that GSK983 acts by inducing a subset of interferon-stimulated genes. PMID- 19187794 TI - Class II HLA-peptide binding prediction using structural principles. AB - The precise prediction of class II human leukocyte antigen (HLA) peptide binding finds application in epitope design for the development of vaccines and diagnostics of diseases associated with CD4+ T-cellular immunity. HLA II binding peptides have an extended conformation at the binding groove unlike class I. This increases peptide binding combinations of varying length at the groove, having an eventual effect in the host immune response to infectious agents. Here we describe the development of a prediction model using information gleaned from HLA II-peptide (HLA II-p) structural data. We created a manually curated dataset of 15 HLA II-p structural complexes from Protein databank (PDB). The dataset was used to develop virtual binding pockets for accommodating HLA-II-specific short peptides. The binding of peptides to the virtual pockets is estimated using the Q matrix (a quantitative matrix based on amino acid residue properties). Internal cross-validation of the model using the 15 HLA II-p structural complexes produced an accuracy of 53% with a sensitivity of 53%. The model was further evaluated using a dataset of 3676 class II-specific peptides consisting of 1188 binders and 2488 nonbinders derived from MHCBN (a database of HLA binders and nonbinders). The model produced an accuracy of 53% with 70.8% specificity and 27.6% sensitivity. The positive predictive value (PPV) was 62% and the negative predictive value (NPV) 58%. A 62% PPV suggests that the model fairly predicts a good number of binders among predicted binders and thus that the success rate among predicted binder for further verification is good. The described model is simple and rapid, with large HLA allele coverage representing the sampled global population, despite weak prediction accuracy. The ability of the model to predict a wide array of defined class II alleles is found to be applicable for proteome wide scanning of parasitic genomes. PMID- 19187795 TI - Recombinant attenuated Salmonella harboring 4-1BB ligand gene enhances cellular immunity. AB - OBJECTIVE: To transfect antigen presenting cells (APCs) with 4-1BB ligand DNA by attenuated Salmonella enterica serovar Typhimurium in vivo, and to observe the effects of ectogenous 4-1BBL on the immune functions of infected rats. METHODS: Attenuated Salmonella typhimurium (vaccine strain) carrying plasmids pIRES2-EGFP 4-1BBL was constructed and used to infect HepG2 hepatoma cells. The expression of reporter gene, green fluorescent protein (GFP) and rat 4-1BBL in the transfected cells was detected by double-immunofluorescence staining. Rats were fed with the recombinant bacteria intragastrically on three occasions in 2 weeks, and were then sacrificed. The transcription and expression of GFP and 4-1BBL genes in splenocytes were measured by RT-PCR and flow cytometry. The phenotypes of T cells in peripheral blood and splenocytes were determined by flow cytometry. The content of IFN-gamma in the cultural supernatant of splenocytes stimulated by PHA was measured by ELISA. RESULTS: The recombinant bacteria harboring 4-1BBL had the same invasive abilities as the original bacteria, and it was able to deliver exogenous genes into HepG2 cells, where the GFP and 4-1BBL were successfully expressed. There were significant upregulations of CD3(+)CD8(+) T cells (P=0.018) and CD3(+)CD25(+) T cells (P=0.019) in the peripheral blood cells as well as CD3(+)CD8(+) T cells (P=0.022), and CD3(+)CD25(+) T cells (P=0.008) in splenocytes of the infected rats. The rats had more 4-1BBL expression detected in the spleen. IFN-gamma released by PHA-stimulated splenocytes increased significantly by the recombinant bacteria as compared with controls (P=0.002). CONCLUSION: Salmonella serovar Typhimurium containing 4-1BBL can transfect target genes into antigen presenting cells in vivo, and the expression of exogenous 4 1BBL enhances cellular immunity markedly. PMID- 19187796 TI - Early replication timing of the chicken alpha-globin gene domain correlates with its open chromatin state in cells of different lineages. AB - The vertebrate alpha-globin gene domain is an open chromatin domain overlapping a neighboring house-keeping gene. The tissue-specific cluster of alpha-globin genes and the overlapping housekeeping gene share the same replication origin. We have studied the replication timing of chicken alpha-globin genes in cells of different lineages using the FISH-based approach and found that alpha-globin genes replicate early both in erythroid and in non-erythroid cells, i.e. regardless of their transcriptional activity. Early replication timing of chicken alpha-globin genes in cells of different lineages was in good correlation with the open chromatin configuration of the alpha-globin gene domain in both erythroid and non-erythroid cells. We propose that active transcription of the housekeeping gene overlapping the alpha-globin gene domain enables an access of Origin Recognition Complex (ORC) proteins to the replication origin resulting in early replication of alpha-globin genes even in non-erythroid cells. PMID- 19187797 TI - Impaired striatal D2 receptor function leads to enhanced GABA transmission in a mouse model of DYT1 dystonia. AB - DYT1 dystonia is caused by a deletion in a glutamic acid residue in the C terminus of the protein torsinA, whose function is still largely unknown. Alterations in GABAergic signaling have been involved in the pathogenesis of dystonia. We recorded GABA- and glutamate-mediated synaptic currents from a striatal slice preparation obtained from a mouse model of DYT1 dystonia. In medium spiny neurons (MSNs) from mice expressing human mutant torsinA (hMT), we observed a significantly higher frequency, but not amplitude, of GABAergic spontaneous inhibitory postsynaptic currents (sIPSCs) and miniature currents (mIPSCs), whereas glutamate-dependent spontaneous excitatory synaptic currents (sEPSCs) were normal. No alterations were found in mice overexpressing normal human torsinA (hWT). To identify the possible sources of the increased GABAergic tone, we recorded GABAergic Fast-Spiking (FS) interneurons that exert a feed forward inhibition on MSNs. However, both sEPSC and sIPSC recorded from hMT FS interneurons were comparable to hWT and non-transgenic (NT) mice. In physiological conditions, dopamine (DA) D2 receptor act presynaptically to reduce striatal GABA release. Of note, application of the D2-like receptor agonist quinpirole failed to reduce the frequency of sIPSCs in MSNs from hMT as compared to hWT and NT mice. Likewise, the inhibitory effect of quinpirole was lost on evoked IPSCs both in MSNs and FS interneurons from hMT mice. Our findings demonstrate a disinhibition of striatal GABAergic synaptic activity, that can be at least partially attributed to a D2 DA receptor dysfunction. PMID- 19187798 TI - Insights into liaison between antiepileptic drugs and bone. AB - The adverse effect on bone caused by chronic anticonvulsant therapy causes multiple abnormalities in calcium and bone metabolism, varying from bone turnover, without significant loss of cortical or trabecular bone, to osteopenia/osteoporosis and to osteomalacic disorder. The studies conducted to date have documented anticonvulsant bone disease as a state of increased bone remodeling. With the newer antiepileptic drugs (AEDs) gaining importance and starting to replace conventional medicines, it may be appropriate to compare them with the conventional AEDs and to examine their impact on multiple aspects of bone health. This review focuses on the status of the bony effects of AEDs. PMID- 19187799 TI - Decision support methods for the detection of adverse events in post-marketing data. AB - Spontaneous reporting is a crucial component of post-marketing drug safety surveillance despite its significant limitations. The size and complexity of some spontaneous reporting system databases represent a challenge for drug safety professionals who traditionally have relied heavily on the scientific and clinical acumen of the prepared mind. Computer algorithms that calculate statistical measures of reporting frequency for huge numbers of drug-event combinations are increasingly used to support pharamcovigilance analysts screening large spontaneous reporting system databases. After an overview of pharmacovigilance and spontaneous reporting systems, we discuss the theory and application of contemporary computer algorithms in regular use, those under development, and the practical considerations involved in the implementation of computer algorithms within a comprehensive and holistic drug safety signal detection program. PMID- 19187800 TI - Seminars in Nuclear Medicine. The role of nuclear medicine techniques in the evaluation of infectious disease (part II). Letter from the editors. PMID- 19187801 TI - Fever of unknown origin. AB - Fever of unknown origin (FUO) often is defined as a fever greater than 38.3 degrees C on several occasions during at least 3 weeks with uncertain diagnosis after a number of obligatory tests. In general, infection accounts for approximately one-fourth of cases of FUO, followed by neoplasm and noninfectious inflammatory diseases. No diagnosis is reached in up to 50% of cases. Scintigraphic methods, such as (67)Ga-citrate, labeled leukocytes, and (18)F fluorodeoxyglucose (FDG) positron emission tomography (PET), are often used in the diagnosis of FUO. In FUO, FDG-PET appears to be of great advantage because malignancy, inflammation, and infection can be detected. FDG-PET does not seem to contribute to a final diagnosis in patients with normal erythrocyte sedimentation rate and C-reactive protein. Image fusion combining PET and computed tomography facilitates anatomical localization of increased FDG uptake and better guiding for further diagnostic tests to achieve a final diagnosis. In conclusion, the body of evidence on the utility of FDG-PET in patients with FUO is growing and FDG-PET will probably become the preferred diagnostic procedure, especially when a definite diagnosis cannot easily be achieved. Because of favorable characteristics of FDG-PET, conventional scintigraphic techniques may be replaced by FDG-PET in institutions in which PET is available. PMID- 19187802 TI - Opportunistic infection and nuclear medicine. AB - Immunosuppression as a result of T- or B-cell dysfunction can be associated with a variety of illnesses as a result of the underlying disease or treatment causing the dysfunction, infection or, indeed, therapy. Immunodeficiency may be primary or secondary. Secondary causes of immune deficiency are more common and more frequently encountered during imaging. Immune deficiency can arise in patients with conditions such as leukemia and lymphoma; from infectious agents such as the human immunodeficiency virus (HIV); from the administration of drugs, including chemotherapy agents and steroids; and as a result of metabolic diseases such as renal failure and diabetes. A condition that often presents challenges in the interpretation of abnormal uptake within radionuclide imaging is the patient with HIV infection. This has been compounded in some ways by the introduction of highly active antiretroviral therapy and the advent of the immune reconstitution inflammatory syndrome. Imaging abnormalities are found in association with the underlying disease, eg, lymphoma, HIV which, on occasion, may be difficult to separate from an opportunistic infection. The primary value of radionuclide imaging and in particular (18)F-fluorodeoxyglucose-positron emission tomography is to rapidly establish the probable site of disease to direct biopsy or aspiration so that the underlying pathology can be confirmed. The value of single photon emission computed tomography and positron emission tomography has been enhanced by the introduction of hybrid imaging so that the computed tomography element of the scan localizes the site of disease more accurately than imaging without the computed tomography. Interest in monitoring response to treatment of infection is increasing but care has to be taken as inflammatory uptake attributable to immune reconstitution inflammatory syndrome can be similar to a worsening of infective uptake and this can lead to misinterpretation of the effect of treatment. It is important for the imager to be aware of the effects of underlying treatments on functional imaging and therefore to have a full history of the disease and the drug treatments that the patient is taking. PMID- 19187803 TI - Cardiovascular infection and inflammation. AB - Infectious and inflammatory processes, in particular those involving the cardiovascular system, are not a story of the past. New aggressive pathogens are responsible for difficult-to-treat infections. Present-day problems such as diabetes are associated with frequent and at times severe infectious processes, with high morbidity related to the disease or to available therapeutic options. Infections involving the heart or vessels pose clinical challenges in diagnosing and planning the most appropriate therapeutic strategy. Inflammatory processes are diagnosed more frequently today, acknowledged as pathologic conditions of high clinical significance, and much is invested in developing efficient therapeutic options. Nuclear medicine procedures are an important component of the evaluation armamentarium used in patients with suspected or confirmed infectious and inflammatory processes. Their role relies on the strength of noninvasive scintigraphic imaging tests that provide functional and metabolic information early during the course of the disease. Drawbacks of nuclear medicine procedures related to either the use of specific radiolabeled tracers or to their rather low resolution are overcome to a large extent during the last decade by the introduction of hybrid positron emission tomography/computed tomography and single-photon emission computed tomography/computed tomography imaging devices. Initial validated results regarding the role of nuclear medicine and of hybrid imaging using various radiotracers in the evaluation of cardiovascular infections and inflammatory processes are emerging over the last few years. They indicate the potentially important role of these modalities for early and precise diagnosis, in defining the whole extent of disease, for individualized treatment tailoring and for monitoring response to treatment. Attention needs to be given to match the appropriate imaging test and radiolabeled agent to the clinical question at hand. Nuclear medicine in general and hybrid imaging procedures in particular will redefine in future the diagnostic and therapeutic capabilities in patients with suspected or known infections and inflammations of the cardiovascular system. PMID- 19187804 TI - Recent trends in soft-tissue infection imaging. AB - This article discusses the current techniques and future directions of infection imaging with particular attention to respiratory, central nervous system, abdominal, and postoperative infections. The agents currently in use localize to areas of infection and inflammation. An infection-specific imaging agent would greatly improve the utility of scintigraphy in imaging occult infections. The superior spatial resolution of (18)F-fluorodeoxyglucose positron emission tomography ((18)F-FDG-PET) and its lack of reliance on a functional immune system, gives this agent certain advantages over the other radiopharmaceuticals. In respiratory tract infection imaging, an important advancement would be the ability to quantitatively delineate lung inflammation, allowing one to monitor the therapeutic response in a variety of conditions. Current studies suggest PET should be considered the most accurate quantitative method. Scintigraphy has much to offer in localizing abdominal infection as well as inflammation. We may begin to see a gradual increase in the usage of (18)F-FDG-PET in detecting occult abdominal infections. Commonly used modalities for imaging inflammatory bowel disease are scintigraphy with (111)In-oxine/(99m)Tc-HMPAO labeled autologous white blood cells. The literature on central nervous system infection imaging is relatively scarce. Few clinical studies have been performed and numerous new agents have been developed for this use with varying results. Further studies are needed to more clearly delineate the future direction of this field. In evaluating the postoperative spine, (99m)Tc-ciprofloxacin single-photon emission computed tomography (SPECT) was reported to be >80% sensitive in patients more than 6 months after surgery. FDG-PET has also been suggested for this purpose and may play a larger role than originally thought. It appears PET/computed tomography (CT) is gaining support, especially in imaging those with fever of unknown origin or nonfunctional immune systems. Although an infection-specific agent is lacking, the development of one would greatly advance our ability to detect, localize, and quantify infections. Overall, imaging such an agent via SPECT/CT or PET/CT will pave the way for greater clinical reliability in the localization of infection. PMID- 19187805 TI - Functional imaging of inflammatory diseases using nuclear medicine techniques. AB - Molecular imaging with positron emission tomography (PET) and single-photon emission computed tomography (SPECT) is increasingly used to diagnose, characterize, and monitor disease activity in the setting of inflammatory disorders of known and unknown etiology. These disorders include sarcoidosis, atherosclerosis, vasculitis, inflammatory bowel disease (IBD), rheumatoid arthritis (RA), and degenerative joint disease. Gallium-67 ((67)Ga) citrate, labeled leukocytes with technetium-99m ((99m)Tc) or indium-111 ((111)In), and (18)F-fluorodeoxyglucose (FDG) represent the most widely used radiopharmaceutical agents. However, other preparations, such as labeled murine monoclonal antigranulocyte antibodies and labeled human polyclonal nonspecific immunoglobulin G, chemotactic peptides, interleukins, chemokines, and liposomes, have been used to image inflammation. Also, (99m)Tc nanocolloid scintigraphy has been found to be suitable for bone and joint diseases, especially RA. Among the single photon emitting imaging agents, the recommended radiotracer for abdominal inflammation has been (99m)Tc-hexamethylpropylene amine oxime (HMPAO)-labeled leukocytes. During the last several years, FDG-PET imaging has been shown to have great value for the detection of inflammation and has become the centerpiece of such initiatives. This very powerful technique will play an increasingly important role in the management of patients with inflammatory conditions. FDG PET can provide valuable information in patients with pulmonary and extrapulmonary sarcoidosis, and is a useful tool for testing the efficacy of various treatments. FDG-PET combined with computed tomography holds great promise for assessing atherosclerosis of the large arteries. This modality is very sensitive in detecting large-vessel vasculitis and can be used to monitor the disease course. FDG-PET is also being used to study the inflamed synovial joints both in the experimental and clinical settings, especially for the investigation and management of RA and degenerative joint disease. This technique also has the potential to become the imaging modality of choice in assessing IBD, replacing radiolabeled autologous leukocyte imaging in this setting. Detection of inflammation in the lungs and airways may improve our knowledge about a multitude of disorders that affect these structures. Therefore, functional imaging, led by FDG-PET imaging, is likely to play an increasingly critical role in assessing inflammatory disorders of known and unknown etiologies, and will improve their management immensely in the future. PMID- 19187806 TI - Radioimmunotherapy of infectious diseases. AB - The need for novel approaches to treat infectious diseases is obvious and urgent. This situation has renewed interest in the use of monoclonal antibodies (mAbs) to treat infectious diseases. During the last 5 years, radioimmunotherapy (RIT), a modality developed for cancer treatment, has been successfully adapted for the treatment of experimental fungal (C. neoformans and H. capsulatum), bacterial (S. pneumoniae and B. anthracis), and viral (HIV-1) infections. RIT produced none or only transient hematological toxicity in experimental animals. Investigation of radiobiological mechanisms of RIT of infections showed that microbial cells are killed by both "direct-hit" and "cross-fire" radiation. mAbs radiolabeled with either alpha- or beta-emitters stimulated apoptosis-like cell death, whereas only mAbs radiolabeled with alpha-emitter (213)Bi also decreased the metabolic activity of microbial cells. The success of this approach in laboratory studies, combined with earlier nuclear medicine experience in preclinical and clinical studies using radiolabeled organism-specific antibodies for imaging of infections, provides encouragement for the feasibility of therapeutically targeting microbes with labeled antibodies. We envision that first the organism specific mAbs will be radiolabeled with imaging radionuclides such as (99m)Tc or (111)In to localize the sites of infection with single-photon emission computed tomography, followed by RIT with (188)Re- or (90)Y-labeled mAb, respectively. Also, immuno-position emission tomogrpahy might be used to image infection before treatment if such positron-emitting radionuclides as (86)Y (matching pair for (90)Y) or (124)I (matching pair for (131)I) are available. It might be possible to create a so-called "pan-antibody" that would recognize an antigen shared by a particular class of human pathogens such as fungi, for example. The availability of such antibodies would eliminate the necessity of having antibodies specific for each particular microorganism and would enormously enhance the development of RIT of infectious diseases. PMID- 19187807 TI - Clinical issues with bisphosphonate therapy for osteoporosis. Introduction. PMID- 19187808 TI - Efficacy of bisphosphonates in reducing fracture risk in postmenopausal osteoporosis. AB - Bisphosphonates have been available for more than a decade. Currently, 4 bisphosphonates--alendronate, risedronate, ibandronate, and zoledronic acid--are approved in the United States. Alendronate and risedronate are oral agents, ibandronate is available in oral and intravenous formulations, and zoledronic acid is an intravenous drug. This review summarizes results from pivotal clinical trials in which these bisphosphonates have been shown to reduce risk for osteoporotic fractures. Also reviewed are results of "bridging" studies designed to demonstrate the comparable efficacy of less frequent dosing regimens to increase bone mineral density and to reduce bone turnover. Compared with placebo controls, all 4 approved bisphosphonates reduce the relative risk of new vertebral fractures in women with postmenopausal osteoporosis. Alendronate, risedronate, and zoledronic acid reduce the relative risk of new nonvertebral and hip fractures. Clinical trial extensions of up to 10 years with alendronate and 7 years with risedronate have shown that efficacy is maintained during long-term treatment. PMID- 19187809 TI - Safety of bisphosphonates in the treatment of osteoporosis. AB - In this review 4 experts consider the major safety concerns relating to bisphosphonate therapy for osteoporosis. Specific topics covered are skeletal safety (particularly with respect to atypical fractures and delayed healing), gastrointestinal intolerance, hypocalcemia, acute-phase (i.e., postdose) reactions, chronic musculoskeletal pain, renal safety, and cardiovascular safety (specifically, atrial fibrillation). PMID- 19187810 TI - Impact of osteoporosis treatment adherence on fracture rates in North America and Europe. AB - Fragility fractures associated with osteoporosis constitute a significant public health concern. Clinical trials have shown that a variety of agents- bisphosphonates, raloxifene, calcitonin, hormone replacement therapy, teriparatide, and strontium ranelate--can reduce the risk of osteoporosis-related fragility fractures. However, low levels of compliance and persistence in the real-life setting mean that efficacy benefits observed in clinical trials with these agents may not translate into equivalent effectiveness in daily practice. The aim of this review is to provide a comprehensive evaluation of compliance and persistence data from retrospective/observational studies, with particular reference to studies that consider the effects on fracture rates. PubMed of the National Center for Biotechnology Information (NCBI) and Web of Science databases were searched for publications detailing observational or retrospective analyses of adherence, compliance, and persistence with osteoporosis therapies. In addition, authors provided relevant studies that were not retrieved using the search criteria. In total, 17 unique publications were identified. Analysis of the publications indicated that low compliance and persistence rates for osteoporosis therapies in the real-life setting result in increased rates of fragility fractures. The results emphasize the importance of good treatment compliance and persistence with osteoporosis therapies in order to achieve a significant therapeutic benefit and thereby reduce the burden that osteoporosis and associated fractures place on individuals and healthcare systems. PMID- 19187811 TI - Osteonecrosis of the jaw and the role of bisphosphonates: a critical review. AB - Osteonecrosis of the jaw (ONJ), a condition characterized by necrotic exposed bone in the maxillofacial region, has been reported in patients with cancer receiving bisphosphonate therapy, and rarely in patients with postmenopausal osteoporosis or Paget disease of bone receiving such therapy. In the absence of a uniform definition, the American Academy of Oral and Maxillofacial Surgeons (AAOMS), the American Society for Bone and Mineral Research (ASBMR), and other groups have established similar diagnostic criteria for bisphosphonate-related ONJ, which is more commonly reported in patients with advanced malignancies with skeletal metastases who receive higher doses, and is more rarely reported in patients with osteoporosis and Paget disease who receive lower doses. However, a critical review of the literature reveals that the etiology of ONJ remains unknown, and to date no direct causal link to bisphosphonates has been established. Despite an increased awareness of ONJ and recent improvements in preventive strategies, patients and physicians alike continue to express concern about the potential risks of bisphosphonate treatment in both oncologic and nononcologic settings. Although much remains to be learned about this condition, including its true incidence in various patient populations, its pathophysiology, and optimal clinical management, evidence to date suggests that the positive benefits of bisphosphonates in patients with malignant bone disease, osteoporosis, or Paget disease outweigh the relatively small risk of ONJ. PMID- 19187812 TI - Sleep hygiene practices of good and poor sleepers in the United States: an internet-based study. AB - This Internet-based investigation assessed the frequency of multiple sleep hygiene practices in 128 good sleepers and 92 poor sleepers (mean age = 41.6, SD = 12.8, 61.8% women). The Pittsburgh Sleep Quality Index was used to measure sleep quality (< 5 = good sleep, > 7 = poor sleep). Although sleep hygiene practices were generally good, poor sleepers showed increased cognitive activity in the bed, even after controlling for global indices of depression and anxiety. Poor sleepers also reported statistically significant increases in excessive noise in the bedroom, uncomfortable nighttime temperature, and activities that were exciting, emotional, or demanded high concentration near bedtime. Future studies should further investigate the prevalence of these variables and their potential impact on sleep quality. PMID- 19187813 TI - Change processes in residential cognitive and interpersonal psychotherapy for social phobia: a process-outcome study. AB - The purpose of this study was to test cognitive and interpersonal models for improving social phobia. Eighty patients with social phobia were randomized to 10 week residential cognitive (RCT) or residential interpersonal psychotherapy (RIPT). They completed process measures every Thursday and a sub-outcome measure every Monday. The ratings were analyzed with mixed models. Weekly changes in the process variables derived from the cognitive model (self-focus, estimated probability and estimated cost of negative social events, safety behaviors) predicted subsequent weekly changes in social anxiety. Changes in the interpersonal variable perceived acceptance by others also predicted subsequent changes in social anxiety. On the other hand, changes in social anxiety predicted changes in the four cognitive variables. There were no interactive effects of process with treatment. The cognitive variables decreased during treatment to a similar degree in both treatments. The results indicate that, to reduce social anxiety, therapy should target self-focus, estimated probability and cost of feared social events, safety behaviors, and perceived acceptance by others. The process of improvement may involve positive cycles in that a reduction of social anxiety, in turn, appeared to impact self-focus, probability, cost, and safety behaviors. PMID- 19187814 TI - Generalized anxiety disorder: connections with self-reported attachment. AB - Even though generalized anxiety disorder (GAD) is one of the most common of the anxiety disorders, relatively little is known about its precursors. Bowlby's attachment theory provides a framework within which these precursors can be considered. According to Bowlby, adult anxiety may be rooted in childhood experiences that leave a child uncertain of the availability of a protective figure in times of trouble.Furthermore, adult "current state of mind with respect to attachment" is thought to relate to adult anxiety. Both attachment-related components were assessed with 8 subscales of the Perceptions of Adult Attachment Questionnaire(PAAQ). Clinically severe GAD clients who were about to begin therapy reported experiencing less maternal love in childhood, greater maternal rejection/neglect, and more maternal role-reversal/enmeshment than did control participants.In keeping with a cumulative risk model, risk for GAD increased as indices of poor childhood attachment experience increased. GAD clients, in contrast to controls,also reported greater current vulnerability in relation to their mothers as well as more difficulty accessing childhood memories. Logistic regression analyses revealed that elevations on PAAQ subscales could significantly predict GAD vs.non-GAD status. Results and the implications for advancing the theory and treatment of GAD are discussed. PMID- 19187815 TI - Correlates of functional impairment in treatment-seeking survivors of mass terrorism. AB - This study sought to identify variables associated with functional impairment in persons exposed to terrorism. A sample of adults who sought treatment for psychological distress related to the 2001 World Trade Center attack completed standardized self-report measures of PTSD symptoms, expectancies of ability to regulate negative moods, interpersonal problems, and social-occupational impairment. A multiple regression analysis found that PTSD numbing symptoms, beliefs about the ability to regulate negative moods, feelings of social discomfort and expectations of being disliked, income level, and relationship status significantly predicted 58% of the variance in social-occupational impairment. The results suggest that treatments targeting PTSD numbing symptoms as well as maladaptive expectations about social interactions and one's ability to manage negative affect may have utility for persons adversely affected by mass violence. PMID- 19187816 TI - Partner schemas and relationship functioning: a states of mind analysis. AB - Cognitions such as relationship attributions and beliefs,measured by self-report, have been identified as robust correlates of relationship (e.g., marital) satisfaction. This study sought to build on the theory and assessment of cognitions in relationships by evaluating partner schemas,defined in terms of self-ratings and recall of positive and negative partner-descriptive trait adjectives. The States of Mind (SOM; Schwartz & Garamoni, 1989) model was used to examine the ratio of positive to positive-plus-negative partner schemas. In a sample of dating college students (N =94), the ratio of positive to positive-plus negative partner schemas was positively associated with relationship satisfaction and negatively associated with distress-maintaining causal and responsibility attributions for partner behavior;these results remained significant when controlling for depressive symptoms. These results suggest that ratios of positive to negative relationship cognitions, including positive and negative views of the partner, and use of information-processing measures of cognition, may be important topics for continued research in understanding relationship functioning. PMID- 19187817 TI - Augmenting in vivo exposure with fear antagonistic actions: a preliminary test. AB - The current study investigated the efficacy of an exposure augmentation strategy in which the phobic individual is encouraged to enact actions that are in direct opposition to the fear action tendencies associated with acrophobia. Participants (N=88) meeting DSM-IV criteria for specific phobia (acrophobia) were randomized to (a) exposure with oppositional actions (E+OA), (b) exposure only (EO), (c) a credible placebo consisting of pulsed audio-photic stimulation (APS), or (d) a waitlist control (WLC). Treatment consisted of six, 6-min exposure trials. Participants were assessed with questionnaire, behavioral, and physiologic measures at pre- and posttreatment, and at a 1-month follow-up session. Participants receiving E+OA showed significantly greater improvement on behavioral and questionnaire measures than those in the other 3 conditions at both posttreatment and follow-up. Further, whereas treatment improvement generalized to an untrained context for those receiving E+OA, such was not the case for EO- and APS-treated participants. Findings suggest augmenting exposure with oppositional actions may enhance treatment outcome and thus warrant additional investigation with clinical samples. PMID- 19187818 TI - Identification with mainstream culture and preference for alternative alcohol treatment approaches in a community sample. AB - Although various treatment approaches are available for alcohol problems, less than 25% of individuals with alcohol use disorders obtain treatment. The purpose of this study is to evaluate interest in attending alternative alcohol treatments, such as meditation and acupuncture, compared to Alcoholics Anonymous (AA). A community sample of 156 adult participants concerned about their drinking were recruited through flyers and newspaper advertisements to complete a Web based survey assessing identification with mainstream culture, sexual identity, and likelihood to attend alternative alcohol treatments. Participants reported higher likelihood of attending alternative treatments as compared to AA, and lesbian, gay, and bisexual participants (28.2% of the sample) were more likely to attend alternative treatments than heterosexual participants. A series of regression analyses were conducted to test whether the relationship between sexual identity and likelihood to attend alternative treatments was mediated by identification with mainstream culture. Participants who were less strongly identified with mainstream culture, regardless of sexual identity, reported higher likelihood of attending alternative treatments. These findings highlight that, for certain subgroups of the population, alternative treatments for alcohol misuse are appealing and suggest the need for future research testing the efficacy of alternative treatments for alcohol problems. PMID- 19187819 TI - Group cognitive behavior therapy for chronic posttraumatic stress disorder: an initial randomized pilot study. AB - Individuals with posttraumatic stress disorder (PTSD) related to a serious motor vehicle accident were randomly assigned to either group cognitive behavioral treatment(GCBT) or a minimum contact comparison group (MCC).Compared to the MCC participants (n=16), individuals who completed GCBT (n=17) showed significant reductions in PTSD symptoms, whether assessed using clinical interview or a self report measure. Among treatment completers, 88.3% of GCBT participants did not satisfy criteria for PTSD at posttreatment assessment, relative to31.3% of the MCC participants. Examination of anxiety,depression, and pain measures did not show a unique advantage of GCBT. Treatment-related gains were maintained over a 3 month follow-up interval. Patients reported satisfaction with GCBT, and attrition from this treatment was comparable with individually administered CBTs.Results are discussed in light of modifications necessitated by the group treatment format, with suggestions for future study of this group intervention. PMID- 19187820 TI - Personality, problem solving, and adolescent substance use. AB - The major aim of this study was to examine the role of social problem solving in the relationship between personality and substance use in adolescents. Although a number of studies have identified a relationship between personality and substance use, the precise mechanism by which this occurs is not clear. We hypothesized that problem-solving skills could be one such mechanism. More specifically, we sought to determine whether problem solving mediates, moderates, or both mediates and moderates the relationship between different personality traits and substance use. Three hundred and seven adolescents were administered the Substance Use Profile Scale, the Social Problem-Solving Inventory-Revised, and the Personality Experiences Inventory to assess personality, social problem solving ability, and substance use, respectively. Results showed that the dimension of rational problem solving (i.e., effective problem-solving skills) significantly mediated the relationship between hopelessness and lifetime alcohol and marijuana use. The theoretical and clinical implications of these results were discussed. PMID- 19187821 TI - Trabeculectomy: more effective at causing cataract surgery than lowering intraocular pressure? PMID- 19187822 TI - The singapore 5-Fluorouracil trabeculectomy study: effects on intraocular pressure control and disease progression at 3 years. AB - PURPOSE: To report 3-year results of a randomized, controlled trial comparing the use of a single application of 5-fluorouracil (5-FU) with placebo in trabeculectomy surgery. DESIGN: Prospective, randomized, double-blinded treatment trial. PARTICIPANTS: Two hundred forty-three Asian patients with primary open angle or primary angle-closure glaucoma undergoing primary trabeculectomy. METHODS: One eye of each patient was randomized to receive either intraoperative 5-FU or normal saline (placebo) during trabeculectomy. MAIN OUTCOME MEASURES: Primary outcome measure was the level of intraocular pressure (IOP). Secondary outcomes were progression of visual field loss, rates of adverse events, and interventions after surgery. RESULTS: Of the 288 eligible patients, 243 were enrolled and 228 completed 3 years follow-up; 120 patients received 5-FU and 123 received placebo. Trial failure, according to predefined IOP criteria, was lower in the 5-FU group compared with the placebo group, although the difference was only significant with a failure criterion of IOP >17 mmHg (P = 0.0154). There was no significant difference in progression of optic disc and/or visual field loss over 36 months between 5-FU and placebo (relative risk [RR], 0.67; 95% confidence interval [CI], 0.34-1.31; P = 0.239). Uveitis occurred more often in the 5-FU treated group (14/115 [12%] vs 5/120 [4%]; P = 0.032). CONCLUSIONS: This is the first masked, prospective, randomized trial reporting the effect of adjunctive 5 FU in trabeculectomy surgery in an East Asian population. The trial shows that an increased success rate can be achieved for several years after a single intraoperative treatment with 5-FU. We conclude that 5-FU is relatively safe and can be routinely used in low-risk East Asian patients. FINANCIAL DISCLOSURE(S): The authors have no proprietary or commercial interest in any materials discussed in this article. PMID- 19187823 TI - Coding variant I62V in the complement factor H gene is strongly associated with polypoidal choroidal vasculopathy. AB - PURPOSE: To investigate whether variants in the complement factor H (CFH) gene are associated with polypoidal choroidal vasculopathy (PCV). DESIGN: Cross sectional study. PARTICIPANTS: A case-control group of 130 PCV subjects and 173 unrelated controls. METHODS: We conducted an association analysis between CFH variants and PCV in a Japanese population, genotyping 12 tag single nucleotide polymorphisms (SNPs)-including rs3753394, rs800292 (I62V), and rs1061170 (Y402H) that are highly representative of the common genetic variation in the CFH region. Genotyping was performed using TaqMan technology. MAIN OUTCOME MEASURES: Allele and haplotype frequencies of the CFH variants. RESULTS: A highly significant association with PCV was observed across the CFH region. The strongest association was observed at I62V (P = 1.7 x 10(-7)). Six other SNPs (rs3753394, rs6680396, rs1410996, rs2284664, rs1329428, and rs1065489) also showed significant association (10(-3) < P < 10(-6)). These associations became nonsignificant after accounting for rs800292 in a conditional logistic regression analysis. A significant omnibus haplotype association was detected in the entire CFH region (omnibus P = 1.6 x 10(-5) at 7 degrees of freedom). Conditional haplotype-based likelihood ratio tests revealed that the significant omnibus haplotype association disappeared when it was estimated conditional on I62V (omnibus P = 0.20, 6 degrees of freedom, post-I62V dependency), whereas the omnibus haplotype association remained significant when it was estimated conditional on any SNP other than I62V. These findings indicate that multiple observed effects were caused by linkage disequilibrium with I62V, and that this variant fully accounts for the association signals observed at the set of SNPs examined at this locus. CONCLUSIONS: The present study provides evidence that the complement pathway plays a substantial role in the pathogenesis of PCV. The nonsynonymous variant I62V is a plausible candidate for a causal polymorphism leading to the development of PCV, given its potential for functional consequences on the CFH protein and our own statistical evidence. FINANCIAL DISCLOSURE(S): The authors have to proprietary or commercial interest in any materials discussed in this article. PMID- 19187824 TI - Retinal vascular caliber and macular telangiectasia type 2. AB - OBJECTIVE: To examine the relationship of retinal vascular caliber to macular telangiectasia (MT) type 2. DESIGN: Case-control study. PARTICIPANTS: Patients with MT aged 18 years and older were identified from Australian sites of the multicenter Macular Telangiectasia Project. Three controls per case were selected from participants of the Australian Diabetes, Obesity and Lifestyle study, matched according to age and diabetes status. METHODS: Baseline ophthalmic examinations of cases included assessment of best corrected visual acuity, fluorescein angiography, autofluorescence imaging, optical coherence tomography, and retinal photography. Retinal vascular caliber of cases and controls were measured from optic disc-centered digital retinal images by a computer-assisted method. MAIN OUTCOME MEASURES: MT, central retinal arteriolar, and venular caliber. RESULTS: There were 55 cases and 170 controls. After controlling for diastolic blood pressure, total cholesterol, high-density lipoprotein cholesterol, triglycerides, hypertension, fasting plasma glucose, and glycosylated hemoglobin, each standard deviation (SD) increase in retinal arteriolar caliber was associated with a 2-fold higher odds of MT (odds ratio [OR] 2.18; 95% confidence interval [CI], 1.50-3.18). Similarly, each SD increase in retinal venular caliber was associated with increased odds of having MT (OR 1.87; 95% CI, 1.31-2.67). CONCLUSIONS: This study shows that MT is associated with wider arteriolar and venular caliber, measured outside of the foveal area. Generalized changes in retinal vascular caliber may reflect underlying dysfunction in retinal pericytes or glial cells, and may provide a means to monitor progression of disease. FINANCIAL DISCLOSURE(S): Proprietary or commercial disclosure may be found after the references. PMID- 19187825 TI - Non-necrotizing herpetic vasculitis. PMID- 19187826 TI - Cerebrovascular accidents in ranibizumab. PMID- 19187827 TI - Lamellar keratoplasty. PMID- 19187829 TI - Binocular single vision field. PMID- 19187830 TI - Refractive error, strabismus, and amblyopia. PMID- 19187831 TI - Amblyopia and strabismus prevalence. PMID- 19187834 TI - [semFYC and the new technologies: where are we coming from and where are we going?]. PMID- 19187835 TI - [Do we reach out too much to the primary care population?]. PMID- 19187836 TI - ["Difficult encounters" in primary care clinic: a patient and doctor perspective]. AB - OBJECTIVES: To find out the prevalence and causes of "difficult encounters" (DE) in Primary Care clinics from the perspective of the patients and doctors, and to whom the responsibility and concordance of the DE (CDE) between doctors and patients is attributed. DESIGN: Cross-sectional, descriptive. SETTING: Urban health centre. PARTICIPANTS AND METHOD: Four doctors, their 4 residents and patients who went to their clinics (March-May 2007). The doctors selected the DE with qualitative criteria, the patients by means of a non-validated ad hoc questionnaire. VARIABLES: sex and age of the patients, defined DE, causes and responsibility of the DE, changes in doctor due to unease and a perception of a "difficult professional" (DP) by the patient. RESULTS: There were 415 visits; 352 questionnaires (85% participation); 212 (60.2%) women, mean age (SD) 54.7 (18.2) years. DE prevalence: 8.5% according to doctors (95% CI, 5.8-11.9) (main cause: idiosyncrasy of the patient), the cause of unease always being attributed to the patient; 3.1% according to the patients (95% CI, 1.6-5.5) (main cause: lack of empathy by the doctor), the unease being attributed to the doctor in 81% of cases. CDE insignificant (kappa=0.003). Only 15 patients (4.3%) felt uncomfortable in a visit in the last year; 18 (5.1%) considered a change of doctor due to unease (95% CI, 3.1-8); 53 (15.1%) had changed at some point for this reason (95% CI, 11.3-18.8) and 11 (3.1%) considered their doctor to be DP (95% CI, 1.6-5.5). CONCLUSIONS: The prevalence of DE in our clinics is similar to that reported in other studies, but lower than expected for the patients, with an insignificant CDE. Doctors and patients usually attribute the responsibility of the DE to the other party. Few patients consider their doctor to be a DP. PMID- 19187837 TI - [Difficult doctor-patient encounters? An unavoidable problem?]. PMID- 19187838 TI - [Translation and cultural adaptation of the Spanish version of EQ-5D-Y questionnaire for children and adolescents]. AB - OBJECTIVE: To translate into Spanish and culturally adapt the international version of EQ-5D-Y for measuring the health related quality of life in the child and adolescent population. DESIGN: Translation and adaptation of a questionnaire. SETTING: Spanish primary and secondary schools. MEASURES: Phases: a) transcultural adaptation of the international tool by means of direct translation to Spanish and back-translation to English, b) evaluation of the clarity, acceptability and familiarity of the first version of the questionnaire by means of cognitive interviews in 20 children and adolescents distributed by sex and age. MAIN RESULTS: The Spanish version of the EQ-5D-Y questionnaire was obtained. The interviewed participants indicated the questionnaire as "clear and understandable". The scores in perceived difficulty were less than 2 out of 10 in the 4 first dimensions of the questionnaire, showing a higher difficulty in the anxiety/depression dimension. CONCLUSIONS: The Spanish version of the presented EQ-5D-Y showed to be understandable and adapted to Spanish children and adolescents, and its ease of use makes this questionnaire potentially useful in different fields, such as primary care and paediatrics. PMID- 19187839 TI - [Early detection of addictive behaviour in primary health care. Use of MULTICAGE CAD-4]. AB - OBJECTIVE: To find out the frequency of specific disorders within the general population using primary healthcare relating to the control of impulses with addictive risk. DESIGN: A cross-sectional descriptive study. LOCATION: Urban health centre, Area 9 of the Madrid Health Service (Spain). PARTICIPANTS: A total of 457 patients who came to primary care (PA) were selected by consecutive sampling. METHOD: Self-completion of the MULTICAGE-CAD4 survey (which measure eight scales for the risky behaviour being studied). RESULTS: There was a risk in at least one of the scales in 46.4% of the sample, of which 17.3% (13.7-20.9) showed risky behaviour on the alcohol scale; 9% (6.2-11.7) for other drugs; 4.4% (2.4-6.4) for gambling; 9.4% (6.6-12.2) for addiction to the Internet; 4.8% (2.7 6.7) for addiction to videogames; 18.8% (15.1-22.5) for eating disorders; 16% (12.5-19.4) for compulsive shopping and 5.5% (3.3-7.7) for addiction to sex. The occurrence of addiction to alcohol, gambling and sex were higher for men, while eating disorders were more prevalent in women. CONCLUSIONS: We are facing a widespread set of problems related to addictions that are often hidden; this study shows that these addictions can be detected within primary healthcare by effective screening tools such as MULTICAGE CAD-4, which is both economic, and easy to implement within primary healthcare centres. PMID- 19187840 TI - [Family doctors and psychiatrists and the patient with depression: the need to re adjust health care approaches and organisational dynamics]. AB - OBJECTIVES: To find out the perceptions of family doctors (FD) and psychiatrists (PS) on their inter-relationships in the health care of patients with depression. DESIGN: Qualitative methodology. Exploratory design based on FD and PS discussion groups (DG). SETTING: Primary health care centres (PHCC) and mental health centres (MHC) of the Basque Country Health Service in Vizcaya. PARTICIPANTS AND CONTEXT: A total of 29 FD from 20 PHCC, and 13 PS from 11 MHC, distributed according to the socioeconomic level of the reference population. The meetings were arranged and held in the PC research unit. METHODS: An intentional sample to configure 4 DG of FD and 2 of PS, homogenous as regards socioeconomic level, and heterogeneous as regards the centres they came from, work experience and gender. The meetings were recorded and transcribed and a sociological discourse analysis was made. Triangulation between researchers and results comparison with the participants was carried out. RESULTS: The perceptions and attitudes of the FD and PS differed, in their connections with the patient, expectations and health care context. In both cases they perceived the unsuitability of the real patient as regards the prefixed perceptions of the professional. The increase of the problem and its management seemed to be conditioned by a social medium which was uncontrolled, and due to increasing deficiencies in the collaboration between health care levels. CONCLUSIONS: Care of the patient with depression requires diagnostic and treatment approaches centred on the patient and shared by FD and PS. PMID- 19187841 TI - [Sacred encounters: serenity in haste]. PMID- 19187842 TI - [How do we fulfil quality standards in emergency care?]. PMID- 19187843 TI - [Reflections on the health systems of Sao Paulo and Catalonia. Common key elements of both health systems (second part)]. PMID- 19187844 TI - [Assessment of the state of health of primary care doctors in the province of Cadiz]. PMID- 19187845 TI - [Is the control of cardiovascular risk factors in secondary prevention adequate?]. PMID- 19187846 TI - [Influence of gender on the approach to cardiovascular disease in primary care]. PMID- 19187847 TI - [Nurses no, doctors yes. Why?]. PMID- 19187848 TI - [Letter to a young resident]. PMID- 19187849 TI - [Microalbuminuria in hypertensive patients in primary care]. PMID- 19187853 TI - Recommendations for the evaluation of left ventricular diastolic function by echocardiography. PMID- 19187854 TI - Distorted cardiac anatomy: the congenital (R)evolution. PMID- 19187855 TI - It's a small world, after all! PMID- 19187856 TI - Echocardiographic evolution of prosthetic aortic valve endocarditis with aorto left atrial fistula formation. AB - Prosthetic valve endocarditis (PVE) due to Staphylococcus aureus is associated with high morbidity and mortality. Paravalvular abscess formation is a common complication of PVE at the aortic position, but fistula formation is rarely seen. This report describes an 84-year-old patient with staphylococcal aortic PVE. Although paravalvular involvement was diagnosed early, clinical features associated with high operative mortality precluded surgical treatment. Unexpectedly, abscess drainage with aorto-left atrial fistula formation led to the remission of fever and clinical stabilization. Moreover, the echocardiographic evolution of infective endocarditis is described, from the initial lesion (vegetation) through abscess formation to partial valve dehiscence, abscess drainage, and aorto-left atrial fistula formation, by means of transesophageal echocardiography. PMID- 19187857 TI - Atrial fibrillation after open heart surgery: how safe is early conversion without anticoagulation? AB - Postoperative atrial fibrillation (AF) is a common complication after cardiothoracic surgery, not only associated with an increased length of hospital stay but also predisposing patients to thromboembolic complications and stroke. Current guidelines advocate a therapeutic approach for postoperative AF that is fairly similar to the recommendations given for nonsurgical patients who develop AF. With respect to anticoagulation, heparin or oral anticoagulation is generally considered appropriate when the arrhythmia persists > 48 hours. The authors present the case of a patient developing sudden-onset, highly symptomatic AF 9 days after open-heart surgery, requiring pharmacologic treatment with amiodarone and metoprolol. Despite an early conversion to sinus rhythm, the patient subsequently developed a stroke, which echocardiographically was attributed to a highly mobile thrombus in the left atrial appendage. Surprisingly, good left atrial mechanical activity was observed. This raises the question of whether the early restoration of sinus rhythm without anticoagulation is safe in a postoperative setting. PMID- 19187859 TI - White matter lesions in spastic paraplegia with mutations in SPG5/CYP7B1. AB - Hereditary spastic paraplegias (HSPs) are relatively frequent disorders presenting great genetic heterogeneity. The recent identification of mutations in SPG5/CYP7B1 in six autosomal recessive kindred linked to the SPG5 locus on chromosome 8q prompted us to test the relative frequency of SPG5/CYP7B1 variants in 12 families and in sporadic HSP patients by high-resolution melting screening combined with direct sequencing. We present two patients who harbored three mutations (including two novel variants) in SPG5/CYP7B1 and white matter involvement evidenced at brain MRI. In HSP patients in whom no other genes were mutated, screening of SPG5/CYP7B1 seems to have a low diagnostic yield in autosomal recessive (8%) and sporadic (<1%) cases, even in those with complicated clinical features. PMID- 19187862 TI - Hepatologists have addressed these severe complications of liver diseases in many ways, but unfortunately the impact of old and modern therapies on survival once complications of liver disease are manifest, is only minor. Preface. PMID- 19187863 TI - Chronic viral hepatitis as a public health issue in the world. AB - This article will focus on the impact caused by chronic viral hepatitis B and C globally and will discuss public health measures that have to be implemented in order to prevent and control these diseases. Chronic viral hepatitis is a major global public health problem, an important cause of morbidity and mortality from sequelae which include chronic hepatitis, cirrhosis and primary liver cancer. Being a 'silent' disease, the contribution of chronic hepatitis to global morbidity and mortality is generally underestimated. Hepatitis B and C prevention and control should seek to reduce both the incidence of new infections and the risk of chronic liver disease. A comprehensive public health prevention programme should include the prevention and detection of HBV and HCV infections, the diagnosis and control of viral hepatitis related chronic liver disease, conducting surveillance and monitoring the effectiveness of prevention activities, and setting up a research agenda. PMID- 19187864 TI - Prevention of viral hepatitis (B and C) reassessed. AB - As hepatitis B and C share modes of transmission, their combined occurrence is not uncommon, particularly in areas where both viruses are endemic and in individuals at high-risk of parenteral infection. Both viral hepatitis infections form an important global public health problem, responsible for over half a billion chronic infections worldwide. Their distinctive characteristics impact upon their epidemiology, transmission, and the success of the different prevention strategies. Since several decades a safe and effective vaccine has been available to prevent hepatitis B virus (HBV) infection. Universal vaccination is the cornerstone of global HBV control. Despite major success, vaccine uptake is hampered, and increasing efforts are required to eliminate acute and chronic hepatitis B. Unlike hepatitis C and HIV, HBV has not captured sufficient attention from policymakers, advocacy groups, or the general public: a major challenge for the future. Although progress has been made in the development of an hepatitis C vaccine, short-term successes are not expected. Even without a vaccine, successes can be reported in the field of hepatitis C due to e.g. implementation of universal precautionary measures in health-care settings, screening of blood and blood products, and identification and counselling of infected people. Despite important efforts, transmission in injecting drug users is increasing. PMID- 19187865 TI - Diagnosis and management of chronic viral hepatitis: antigens, antibodies and viral genomes. AB - Virological tools, including serological and molecular tools, are needed to diagnose chronic hepatitis B and C infections. They may also be useful to establish their prognosis, but they have found their principal application in guiding treatment decisions and assessing the virological responses to therapy. The goal of chronic hepatitis B therapy is to prevent progression of liver disease. This is achieved if HBV replication is durably abolished or significantly reduced. In HBeAg-positive patients, HBeAg clearance followed by the HBe seroconversion phase can be achieved. In HBeAg-negative patients, long term antiviral suppression of viral replication is needed. The loss of HBsAg, eventually associated with an HBs seroconversion, is the most desirable endpoint of therapy but is rarely achieved. The efficacy endpoint of chronic hepatitis C treatment is the sustained virological response, defined by an undetectable HCV RNA in serum with a sensitive assay 24 weeks after the end of treatment. The HCV genotype and on-treatment viral kinetics can be used to tailor treatment dosages and duration. PMID- 19187866 TI - Immunology of hepatitis B and hepatitis C virus infections. AB - Hepatitis B (HBV) and hepatitis C (HCV) viruses are the two major causes of chronic liver inflammation worldwide. Despite distinct virologic features, both viruses are preferentially hepatotropic, not directly cytopathic, and elicit liver diseases that share several aspects of their natural history. HBV and HCV infections also share some important features of the adaptive antiviral immune response. We describe the innate immune response in the early phase following infection, and how these early events may influence the development of the adaptive immune response in these two important viral infections. The mechanisms by which high levels of viral antigens, liver immunological features, the presence of regulatory T cells and impaired dendritic cell functions may maintain the HBV- and HCV-specific immunological failure, characteristic of chronic hepatitis B and C patients, are also evaluated. PMID- 19187867 TI - Natural history: the importance of viral load, liver damage and HCC. AB - Chronic hepatitis B and hepatitis C virus infections are the major causes of liver disease, hepatocellular carcinoma (HCC) and liver-related mortality worldwide. Among factors known to influence the natural history of viral hepatitis are age at the time of infection, duration of infection, serum alanine aminotransferase (ALT) levels, male sex, alcohol consumption, and coinfections. In hepatitis B, serum HBV DNA concentration emerges as the key factor for predicting the development of liver disease. Even patients with low viraemia seem at increased risk for liver cirrhosis and HCC. Coinfections with hepatitis C, hepatitis D and/or HIV are common and are associated with a more severe liver disease. The course of chronic hepatitis C is variable, but usually fibrosis advances slowly. In addition to the better-known factors- including coinfections with HBV and HIV- progression of liver disease is adversely affected by smoking, hepatic steatosis and insulin resistance. PMID- 19187868 TI - Nucleos(t)ide analogues for hepatitis B virus: strategies for long-term success. AB - Studies in the past decades have shown that active hepatitis B virus (HBV) replication is the key driver of liver injury and disease progression, and thus sustained viral suppression is of paramount importance in the management of chronic HBV infection. The nucleos(t)ide analogues lamivudine, adefovir, entecavir, telbivudine and tenofovir are potent inhibitors of HBV polymerase/reverse transcriptase activity and are highly effective in the suppression of HBV replication, but rarely eliminate the virus. Long-term therapy is usually required to achieve sustained hepatitis B e antigen seroconversion, HBV DNA suppression, ALT normalization and fibrosis reversal. Maintained long term therapy has been demonstrated to significantly lower the rate of hepatic decompensation and development of cirrhosis or hepatocellular carcinoma. However, drug resistance is a serious risk on prolonged nucleos(t)ide analogue therapy, and this poses a critical challenge. Prevention and proper management of drug resistance are crucial to ensure long-term success. PMID- 19187869 TI - Peginterferon for the treatment of chronic hepatitis B in the era of nucleos(t)ide analogues. AB - The practising clinician is currently faced with a number of effective treatment options for chronic hepatitis B, including two formulations of interferon (standard IFN and pegylated IFN) and five nucleos(t)ide analogues (lamivudine, adefovir, entecavir, telbivudine and tenofovir). Treatment strategies can be divided into those aiming for sustained response after discontinuation of therapy and those that need to be maintained by prolonged antiviral therapy. Sustained response is particularly achieved with interferon-based therapy, while treatment maintained response can be achieved with long-term nucleos(t)ide analogue therapy in the majority of patients. Of currently available drugs for the treatment of chronic hepatitis B, PEG-IFN seems to result in the highest rate of off-treatment sustained response after a 1-year course of therapy. Sustained transition to the immune-control phase (inactive HBsAg carrier state) can be achieved in 30-35% of HBeAg-positive patients and 20-25% of HBeAg-negative patients. Loss of HBsAg has been observed in 11% of both HBeAg-positive and HBeAg-negative patients after 3-4 years. Since hepatitis B virus (HBV) genotype is an important predictor of response to PEG-IFN, determination of HBV genotype is essential in patients in whom sustained off-treatment response is pursued. Aiming for sustained response is of particular interest because many HBV-infected patients are in need of antiviral therapy at a young age and may otherwise require indefinite antiviral therapy. PMID- 19187870 TI - Peginterferon and ribavirin in HCV: improvement of sustained viral response. AB - Peginterferon alfa in combination with ribavirin is and will remain for the next years the current standard for treatment of chronic hepatitis C. The new antivirals currently investigated in phase II of III trials may augment the overall response rates but require peginterferon/ribavirin as backbone. The cure rate of peginterferon/ribavirin treatment can be improved by better education of treating physicians to identify and treat conditions which negatively influence the final outcome of therapy. Specific focus is the prevention and/or early treatment of common side effects of therapy including anaemia, cytopenia and depression. In selected patients increasing the dose of peginterferon and/or ribavirin may augment response rates. Measuring the viral response at various time points during treatment allows individualization of treatment duration. Treatment duration may be shortened in patients with undetectable HCV-RNA after 4 weeks of therapy, on the other hand slow virologic responders may benefit from prolonged treatment. PMID- 19187871 TI - Future treatment options for HCV: double, triple, what is the optimal combination? AB - Specifically Targeted Antiviral Therapy against hepatitis C virus (STAT-C) stands for a new era in the treatment of patients with chronic hepatitis C. Results from recent trials with protease and polymerase inhibitors indicate that therapy with a single HCV specific compound will not be sufficient to eradicate hepatitis C virus infection and that combination therapy will be necessary to improve sustained virologic response rates. The search for the optimal combination of STAT-C compounds with peginterferon alfa with or without ribavirin is currently under investigation in several clinical trials. Overall the current studies indicate that peginterferon alfa and ribavirin remain the backbone of antiviral therapy of chronic hepatitis C even in the era of STAT-C. Nevertheless, it can be anticipated that combination of STAT-C compounds with non-overlapping resistance profiles could improve response to antiviral therapy. Promising combinations are protease inhibitors plus nucleoside analogue and non-nucleoside analogue polymerase inhibitors. PMID- 19187872 TI - Hepatocellular carcinoma in viral hepatitis: improving standard therapy. AB - Hepatocellular carcinoma (HCC) is a common malignancy worldwide, and its incidence is increasing in the United States and elsewhere. The prognosis of HCC patients depends not only on tumour stage but also on the background liver function reservoir. Current options for the treatment of HCC are surgical resection, liver transplantation, transcatheter arterial embolization, chemotherapy, and percutaneous ablation therapy. The choice of optimal treatment for individual patients, especially those at an earlier cancer stage, is sometimes controversial. Short-term prognosis of HCC patients has been much improved recently due to advances in early diagnosis and treatment, although long term prognosis is as yet far from satisfactory as indicated by the overall survival at 10 years after apparently curative treatment of only 22-35%. Prevention of HCC recurrence, or tertiary prevention, is one of the most challenging tasks in current hepatology. PMID- 19187873 TI - Liver transplantation in viral hepatitis: prevention of recurrence. AB - End-stage liver disease caused by the hepatitis B and C viruses (HBV and HCV) are major indications for liver transplantation. Outcome depends largely on the prevention of allograft reinfection. The advent of long-term hepatitis B immune globulin administration and the introduction of new antiviral agents were a major breakthrough in the management of these patients. Today, survival after orthotopic liver transplantation (OLT) is similar to that of patients transplanted for HBsAg-negative liver disease, and the risk of recurrence is below 10%. In contrast, HCV reinfection is almost constant and significantly impairs patient and graft survival. Factors that may influence disease severity and consequently progression of HCV graft injury remain unclear. Pre transplantation and prophylactic post-transplantation antiviral treatments are limited by low applicability and poor tolerance. Treatment of established graft lesions with combination therapy gave promising results, with sustained virological response in 25-45% of patients, but indications, modality and duration of treatment should be assessed. PMID- 19187874 TI - Successful aging or aging successfully. PMID- 19187875 TI - Can successful aging and chronic illness coexist in the same individual? A multidimensional concept of successful aging. AB - For the past 4 decades, many studies seeking to develop a definition and identify critical determinants of successful aging have been published. To date, there is still no consensus on a standard definition or measure of "successful aging." Most constructs have been one dimensional; although a few have been multidimensional, none has emerged as standard. The most serious limitation of contemporary successful aging constructs is the undue focus on physiologic aspects of aging. To move beyond this limited perspective that stresses disease and impairment, we postulate that successful aging may coexist with diseases and functional limitations if compensatory psychological and/or social mechanisms are used. With this premise, this article presents a new definition and conceptual framework of successful aging, together with an operational definition (measurement) that delineates our successful aging concept. We discuss how the proposed multidimensional measurement may be used as a screening tool, and address its relevance for health services research and health care delivery. PMID- 19187876 TI - Profile of physicians in the nursing home: time perception and barriers to optimal medical practice. AB - OBJECTIVES: To describe physician medical practice in nursing homes, including actual time spent for routine encounters with nursing home residents and demographic characteristics of the physicians who serve as medical directors; to determine the congruence between actual time spent for routine encounters with nursing home residents and the physician's view of the optimal time; and to identify barriers to providing optimal visits. DESIGN: A mail survey of a national random sample of 200 medical directors of all Medicare-certified nursing facilities using the Dillman Total Design mail survey methodology. PARTICIPANTS: 100 medical directors (50% response rate). MEASUREMENTS: The survey consisted of open- and closed-ended items on the following: the demographic characteristics of the medical director; demographic characteristics of the nursing home; the extent of the medical director's nursing home practice, including the ideal and actual time spent in nursing home visits for 4 common types of visits; and perception of barriers to providing optimum visits in the nursing home. RESULTS: Medical directors were most likely to be primary care physicians, the majority of whom were male; had practiced in long-term care for more than 18 years; were medical directors in 2 facilities; provided, on average, primary care in 4 facilities; spent 31 hours per month in the nursing home with nursing staff; and devoted 44% of their practice to nursing homes. Most, (74%) were members of the American Medical Directors Association (AMDA), 41% were certified medical directors (CMD), 42% had a certificate of added qualification (CAQ) in geriatrics, and only 15% had fellowship training. Reports of actual time spent on 4 common types of nursing home visits were significantly less than optimal visit times, but fellowship-trained physicians reported significantly greater discrepancies between the optimal and actual time spent for the 30- to 60-day reviews and readmissions compared with physicians who were not. A parallel pattern was seen comparing physicians with and without CAQs. Nursing support and accurate/accessible information were recorded as most problematic and reimbursement the least problematic barrier to providing optimal nursing home visits. CONCLUSION: The present study provides a snapshot of current physician practice in US nursing homes. Such information is needed as the debate over the physician's role in the nursing home continues and new policy is framed that will ultimately define the future of medical practice in the nursing home. That 74% of the national survey respondents were members of AMDA suggests that the AMDA membership is representative of the national medical director population. PMID- 19187877 TI - "Sometimes people don't fit in boxes": attitudes toward the minimum data set among clinical leadership in VA nursing homes. AB - OBJECTIVE: To describe attitudes toward the Minimum Data Set (MDS) among Department of Veterans Affairs (VA) nursing home care unit (NHCU) clinical leadership. DESIGN: Cross-sectional online survey using structured (quantitative) and open-ended (qualitative) items. SETTING: Approximately 97 VA Medical Center NCHUs nationwide. PARTICIPANTS: A total of 289 directors of nursing, medical directors, MDS coordinators, nurse managers and other clinical management staff. MEASUREMENTS: Quantitative ratings of the MDS's accuracy, usefulness for initial and ongoing care planning, and utility for quality improvement; content analysis of open-ended items describing perceived causes of inaccuracy and reasons for use/nonuse in care planning and quality improvement. RESULTS: Although quantitative ratings were generally positive, qualitative analysis yielded a number of emergent themes regarding data accuracy, team functioning, timeliness of assessments, and validity of the MDS tool itself. Medical directors were somewhat less positive about the MDS than were other NHCU leadership. Very large and very small facilities were less likely to view the tool as useful and to use it for care planning and quality improvement. CONCLUSIONS: NHCU clinical leadership clearly has a "love-hate" relationship with the MDS. They value information it provides, but identify a number of weaknesses that limit its utility for clinical use. PMID- 19187878 TI - Medication management in assisted living: a national survey of policies and practices. AB - OBJECTIVE: To obtain information about actual medication management practices in assisted living residences (ALRs). DESIGN: An online survey; data were collected and reported as frequencies. Operational definitions were provided for "assist with" and "administration of" medications. SETTING: All 50 states. PARTICIPANTS: Members of the key assisted living professional and provider associations. RESULTS: More than half of ALRs (n = 547) administer medications to 80% to 100% of their residents. Almost half of ALRs use unlicensed assistive personal (UAP)/medication aides to administer medications, as permitted by state regulations. In those states where UAPs are not permitted, unlicensed staff may assist residents with their medications. More than half of ALRs have written policies regarding medication storage and documentation of administration. A slightly smaller percentage of ALRs have policies regarding medication administration by a UAP/med aide, quality improvement programs, and error reporting. As much as 30% of ALRs lack policies regarding drug regimen review and monitoring for adverse drug events (ADEs). CONCLUSION: Intensive education for practitioners about appropriate prescribing is warranted, as is improved methods for assessment of an older adult's ability to safely self-administer medications. The role of Boards of Nursing in every state, and nationally, should take a leadership role in establishing the curriculum, training, competencies, and performance evaluation criteria of UAP/med aides. PMID- 19187879 TI - Atypical antipsychotics and the risk of diabetes in an elderly population in long term care: a retrospective nursing home chart review study. AB - OBJECTIVES: Although atypical antipsychotics (AA) are associated with weight gain and diabetes mellitus (DM) in younger patients, it is not known whether these drugs would have a detrimental effect on weight gain or diabetes in a long-term care elderly population. DESIGN: Retrospective chart review. SETTING: Two community nursing homes (NH). PARTICIPANTS: Charts of 1678 subjects admitted between 2000 and 2006 were screened; data from subjects on AA were analyzed. MEASUREMENTS: DM was defined by diagnosis in the Minimum Data Set (MDS), the prescription of diabetes medications, fasting blood glucose (BG) 126 mg/dL or higher, or random BG 200 mg/dL or higher. Worsening of DM was defined as beginning a medication in those treated with diet alone, or adding an additional medication to those already on antiglycemic therapy. RESULTS: There were 154 subjects on AA, mean age 82.8 +/- 8.0 (SD) years. Although there was no difference in age between the 101 women and the 53 men, there was a greater proportion of women 85 years or older compared with men (57% versus 40%, P = .04). Alzheimer's disease (AD) was diagnosed in 17% and non-AD dementia in 34%. Weight was normal (BMI less than 25 kg/m(2)) in 63%, overweight (BMI higher than 25 but less than 30 kg/m(2)) in 29%, and obese (BMI higher than 30 kg/m(2)) in 8%. Subjects were followed on AA for a median of 13.1 weeks (interquartile range 1.9-41.9). Despite these medications, 32% lost more than 5% of body weight. DM was an admitting diagnosis in 21%. There were 4 new and 5 worsening cases of DM during their stay in the nursing home; however, of these 9 cases, 4 occurred before the institution of AA. There was no increased frequency of weight gain or DM among the various atypical agents. CONCLUSIONS: In an elderly NH population, there was no evidence that short-term use (median 13.1 weeks) of atypical antipsychotic agents was associated with the onset or worsening of DM. PMID- 19187880 TI - Chronic kidney disease, anemia, and the association between chronic kidney disease-related anemia and activities of daily living in older nursing home residents. AB - OBJECTIVES: The aim of this study was to evaluate the potential association between anemia associated with chronic kidney disease (CKD) and the ability for older nursing home (NH) residents with these conditions to perform basic activities of daily living (ADL). DESIGN: This is a multicenter, retrospective, observational study utilizing data abstracted from medical charts, laboratory records, and Minimum Data Set (MDS) records from the 1-year period before the date of data abstraction. SETTING: 24 NHs in the United States. PARTICIPANTS: Older (>or=65 years of age) NH residents with CKD (not receiving dialysis) who were not comatose and were alive as of the day of data abstraction were included in this analysis. MEASUREMENTS: Resident demographics, hemoglobin, and estimated glomerular filtration rate records, and ADL items were abstracted from the medical charts, laboratory records, and MDS. The prevalence of CKD and anemia associated with CKD was evaluated from laboratory records. The relationship between CKD, anemia, and ADLs was assessed through a repeated resampling (bootstrap) analysis. RESULTS: CKD was present in 45.7% of the residents. Of these, 60.5% had anemia. Residents with CKD and anemia were more likely to require limited or extensive assistance in performing ADLs such as bed mobility, transfer, walk in room, walk in corridor, locomotion on unit, dressing, and toilet use. Conversely, a greater proportion of residents with CKD but without anemia required no or slight supervision in performing these ADLs. CONCLUSION: CKD and anemia associated with CKD are common in older NH residents. The data suggest that older NH residents with CKD and anemia require greater assistance in performing ADLs than residents with CKD alone. These data support the importance of evaluating the older NH population for CKD and anemia. As with all retrospective analyses, study limitations must be considered in the interpretation of these results. PMID- 19187882 TI - Impact of a home-based primary care program in an urban Veterans Affairs medical center. AB - OBJECTIVES: Describe how an interdisciplinary home-based primary care program (HBPC) affected hospital and emergency department (ED) use in an urban Veterans Affairs medical center. DESIGN: A retrospective review. SETTING: HBPC of the Washington, DC, Veterans Affairs Medical Center (VAMC-DC). PARTICIPANTS: All HBPC patients enrolled for at least 6 months during the period of January 1, 2001 through December 31, 2002. MEASUREMENTS: Baseline demographic variables (such as age, gender, race, living arrangement, community services used), major medical diagnoses, functional measures (MMSE, ADL, IADL, Braden), advance directives, episodes of ED, and hospital use for the 6-month period before and after HBPC enrollment from the VAMC-DC and discharge status (obtained from electronic medical records). RESULTS: A total of 183 patients were enrolled in HBPC for at least 6 months during the 24-month study period. Ninety-five percent were male with a mean age of 73.6 years (range 36 to 95). Most were African American (71%) and lived with a caregiver (65%). The average number of comorbidities was 6 per patient. At the end of the 2-year study period, 45% remained active in HBPC and 17% had died. Using paired score analysis t tests, patients enrolled in HBPC for 6 months had 43.7% fewer hospital admissions (P = .001) and spent 49.9% fewer days in the hospital (P = .001). The 18.5% reduction in ED visits was not statistically significant (P = .2632). CONCLUSIONS: Use of HBPC for 6 months for frail chronically ill patients in an urban VAMC may be associated with fewer hospital admissions resulting in reduced total hospital days, but no significant change in ED use. PMID- 19187881 TI - Not so black and white: nursing home concentration of Hispanics associated with prevalence of pressure ulcers. AB - OBJECTIVE: To examine the association between the nursing home (NH) concentration of Hispanics and prevalence of pressure ulcer. DESIGN: Secondary data analysis using data from the national repository of the 2000 Minimum Data Set (MDS) and On Line Survey Certification and Reporting (OSCAR) data. We used a multivariate logistic model, with the Huber-White correction to account for clustering of persons within a nursing facility, to examine the association of Hispanic NH concentration with the prevalence of pressure ulcers, after adjusting for resident level characteristics. SETTING: Five states with a high population of Mexican-Americans (California, New Mexico, Arizona, Colorado, and Texas). PARTICIPANTS: A total of 74,343 persons (9.26% black, 11.28% Hispanic, 79.46% non Hispanic white) in a NH located in 1 of these 5 states during the last quarter of 2000. MEASUREMENTS: The prevalence of Stage II-IV pressure ulcers was examined in the last quarter of 2000. Stage II-IV pressure ulcers, resident demographics, and medical illness data were documented by nursing staff on the MDS. RESULTS: Hispanics and non-Hispanic blacks had a higher prevalence of pressure ulcers than non-Hispanic whites (7.60%, 9.71% and 12.10%, respectively). A facility's concentration of Hispanic residents was associated with prevalent pressure ulcers after adjustment for resident characteristics. CONCLUSIONS: Residents in nursing homes in which there is a higher concentration of Hispanic residents are more likely to have a pressure ulcer. PMID- 19187883 TI - Plombage migration outside the thoracic cavity: a complication of tuberculosis treatment. PMID- 19187884 TI - Pressure ulcer PUSH score and traditional nursing assessment in nursing home residents: do they correlate? AB - BACKGROUND: Over a decade ago, the National Pressure Ulcer Advisory Panel (NPUAP, 1997) recommended a new tool, the Pressure Ulcer Scale for Healing (PUSH) tool to document ulcers and monitor the healing process. Yet, traditional nursing observation remains standard practice in chronic care, thus prompting this correlational study between PUSH and traditional documentation of pressure ulcers. METHODS: Data were cross-tabulated through a retrospective chart review of all residents with stage II-IV decubiti at a 672-bed skilled nursing facility, between January 1, 2004, and December 31, 2006. A correlation analysis was performed between the clinical nursing observation, which was based primarily on ulcer size and documented on the weekly decubiti flow sheets, and the weekly PUSH score over a period of at least 2 months. Agreement was assessed using kappa statistics for a 3 x 3 table between the nurse's impression (improved, unchanged, deteriorating) and the change in PUSH score (+1, 0, or -1). RESULTS: In the 370 observations compiled for the 48 residents, the nurses documented improvement in 212 observations (57%). However, of these 212 traditionally assessed "improved" ulcers, there were only 89 (42%) concordant "better" PUSH scores and 99 (47%) received a "no change" PUSH score. Twenty-four (11%) of the 212 actually received a deterioration of the ulcer rate using the PUSH tool. Of the 110 (30%) traditionally assessed as "unchanged" ulcers, only 45 (42%) matched "unchanged" PUSH scores. Finally, for the 48 (13%) traditionally documented "deteriorating" ulcers, there were only 25 (52%) observations in agreement with the "deteriorating" PUSH scores. Overall, in this longitudinal study, the symmetric measures reports indicated very little agreement between the 2 assessment methods (kappa range: 0.007-0.298). CONCLUSION: Although the NPUAP has formally recommended the PUSH tool as the pressure ulcer assessment method of choice, our data indicate that the PUSH does not highly correlate with traditional nursing observation. Further study is required to determine the most accurate assessment method. The adoption of a universally accepted tool, together with rigorous documentation methods, will improve the overall clinical care of chronic patients with pressure ulcers. PMID- 19187885 TI - Threading the needle between compassionate care and fostering dependency: an accurate assessment of disability. PMID- 19187886 TI - Behaviors in advanced dementia. PMID- 19187888 TI - Clinical causes of death of 2372 older persons in a nursing home during 15-year follow-up. PMID- 19187890 TI - Opioids for chronic noncancer pain: prediction and identification of aberrant drug-related behaviors: a review of the evidence for an American Pain Society and American Academy of Pain Medicine clinical practice guideline. AB - Optimal methods to predict risk of aberrant drug-related behaviors before initiation of opioids for chronic noncancer pain and to identify aberrant behaviors after therapy is initiated are uncertain. We systematically reviewed published literature identified through searches of Ovid MEDLINE and the Cochrane databases through July 2008. Diagnostic test characteristics and accompanying confidence intervals were calculated with data extracted from the studies. Four prospective studies evaluated diagnostic accuracy of risk prediction instruments. Two higher-quality derivation studies found that high scores on the Screener and Opioid Assessment for Patients with Pain (SOAPP) Version 1 and the Revised SOAPP (SOAPP-R) instruments weakly increased the likelihood for future aberrant drug related behaviors (positive likelihood ratios [PLR], 2.90 [95% CI, 1.91 to 4.39] and 2.50 [95% CI, 1.93 to 3.24], respectively). Low scores on the SOAPP Version 1 moderately decreased the likelihood for aberrant drug-related behaviors (negative likelihood ratio [NLR], 0.13 [95% CI, 0.05 to 0.34]) and low scores on the SOAPP R weakly decreased the likelihood (NLR, 0.29 [95% CI, 0.18 to 0.46]), but estimates are too imprecise to determine if there is a difference between these instruments. One lower-quality study found that categorization as high risk using the Opioid Risk Tool strongly increased the likelihood for future aberrant drug related behaviors (PLR, 14.3 [95% CI, 5.35 to 38.4]) and classification as low risk strongly decreased the likelihood (PLR, 0.08 [95% CI, 0.01 to 0.62]). Nine studies evaluated monitoring instruments for identification of aberrant drug related behaviors in patients on opioid therapy. One higher-quality derivation study found higher scores on the Current Opioid Misuse Measure (COMM) weakly increased the likelihood of current aberrant drug-related behaviors (PLR, 2.77 [95% CI, 2.06 to 3.72]) and lower scores weakly decreased the likelihood (NLR, 0.35 [95% CI, 0.24 to 0.52]). In 8 studies of other monitoring instruments, diagnostic accuracy was poor, results were difficult to interpret due to methodological shortcomings, or standard diagnostic test characteristics were not reported. Definitions for aberrant drug-related behaviors were not standardized across studies and did not account for seriousness of identified behaviors. No reliable evidence exists on accuracy of urine drug screening, pill counts, or prescription drug monitoring programs; or clinical outcomes associated with different assessment or monitoring strategies. PERSPECTIVE: Evidence on prediction and identification of aberrant drug-related behaviors is limited. Although several screening instruments may be useful, evidence is sparse and primarily based on derivation studies, and methodological shortcomings exist in all studies. Research that performs external validation, uses standardized definitions for clinically relevant aberrant drug-related behaviors, and evaluates clinical outcomes associated with different assessment and monitoring strategies is needed. PMID- 19187889 TI - Clinical guidelines for the use of chronic opioid therapy in chronic noncancer pain. AB - Use of chronic opioid therapy for chronic noncancer pain has increased substantially. The American Pain Society and the American Academy of Pain Medicine commissioned a systematic review of the evidence on chronic opioid therapy for chronic noncancer pain and convened a multidisciplinary expert panel to review the evidence and formulate recommendations. Although evidence is limited, the expert panel concluded that chronic opioid therapy can be an effective therapy for carefully selected and monitored patients with chronic noncancer pain. However, opioids are also associated with potentially serious harms, including opioid-related adverse effects and outcomes related to the abuse potential of opioids. The recommendations presented in this document provide guidance on patient selection and risk stratification; informed consent and opioid management plans; initiation and titration of chronic opioid therapy; use of methadone; monitoring of patients on chronic opioid therapy; dose escalations, high-dose opioid therapy, opioid rotation, and indications for discontinuation of therapy; prevention and management of opioid-related adverse effects; driving and work safety; identifying a medical home and when to obtain consultation; management of breakthrough pain; chronic opioid therapy in pregnancy; and opioid related policies. PERSPECTIVE: Safe and effective chronic opioid therapy for chronic noncancer pain requires clinical skills and knowledge in both the principles of opioid prescribing and on the assessment and management of risks associated with opioid abuse, addiction, and diversion. Although evidence is limited in many areas related to use of opioids for chronic noncancer pain, this guideline provides recommendations developed by a multidisciplinary expert panel after a systematic review of the evidence. PMID- 19187892 TI - History will judge... PMID- 19187891 TI - Research gaps on use of opioids for chronic noncancer pain: findings from a review of the evidence for an American Pain Society and American Academy of Pain Medicine clinical practice guideline. AB - Chronic noncancer pain is common and use of opioids is increasing. Previously published guidelines on use of opioids for chronic noncancer pain have been based primarily on expert consensus due to lack of strong evidence. We conducted searches on Ovid MEDLINE and the Cochrane databases through July 2008 to identify studies that addressed one or more of 37 Key Questions that a multidisciplinary expert panel identified as important to be answered to generate evidence-based recommendations on the use of opioids for chronic noncancer pain. A total of 14 systematic reviews, 38 randomized trials not included in a previously published systematic review, and 13 other studies met inclusion criteria. Almost all of the randomized trials of opioids for chronic noncancer pain were short-term efficacy studies. Critical research gaps on use of opioids for chronic noncancer pain include: lack of effectiveness studies on long-term benefits and harms of opioids (including drug abuse, addiction, and diversion); insufficient evidence to draw strong conclusions about optimal approaches to risk stratification, monitoring, or initiation and titration of opioid therapy; and lack of evidence on the utility of informed consent and opioid management plans, the utility of opioid rotation, the benefits and harms specific to methadone or higher doses of opioids, and treatment of patients with chronic noncancer pain at higher risk for drug abuse or misuse. PERSPECTIVE: Currently, clinical decisions regarding the use of opioids for chronic noncancer pain need to be made based on weak evidence. Research funding priorities need to be set to address these critical research needs if the care of patients with chronic noncancer pain is to improve. PMID- 19187893 TI - Improving visual skills: II-remote assessment via Internet. AB - BACKGROUND: Even though poor readers often have poor visual skills, such as binocular coordination and oculomotor control, students' visual skills are rarely assessed. Computer assessments have the potential to assist in identifying students whose visual skills are deficient. This study compared assessments made by an Internet-based computer orthoptics program with those of an on-site vision therapist. METHODS: Students (N = 41) in grades 1 through 8, reading at least 2 levels below grade, were assessed for visual skill dysfunction (including binocular fusion and tracking ability) by a vision therapist at their school in Wisconsin. The therapist determined whether the student had adequate visual skills based on clinical and behavioral observations. A "remote" investigator located in California determined the adequacy of accommodative facility, tracking, and vergence skills in the same students, based on quantitative progress through the modules of an Internet-based computer orthoptics training program during 3 assessment sessions. RESULTS: The on-site therapist made 33 referrals for possible visual skills training (80%). The remote investigator made 25 referrals (61%), all of which were consistent with referrals made by the on site therapist; thus, no false-positives occurred when using the remote assessment technique. The 8 additional referrals by the therapist were attributed to the ability to observe student behavior during assessment. CONCLUSIONS: Remote assessment of visual skills via an Internet orthoptics program may provide a simple means to detect visual skill problems experienced by poor readers. PMID- 19187894 TI - Two differing presentations of chronic bilateral anterior uveitis. AB - BACKGROUND: Bilateral, recurrent, or chronic anterior uveitis requires a diagnostic evaluation to rule out any systemic cause. An understanding of the possible etiologies and their diagnostic criteria is needed to manage these patients. Treating any systemic cause can decrease the recurrent or chronic nature of the uveitis and favorably alter the course. Many possible systemic conditions are capable of causing anterior uveitis, including sarcoidosis. CASE REPORTS: Two cases of chronic, recurrent, bilateral uveitis are presented. Both were evaluated for any systemic etiology. After extensive systemic workups, the first case had no identifiable systemic etiology, whereas the second case was associated with systemic sarcoidosis. CONCLUSION: Although a medical workup may be necessary, it will not always lead to a systemic diagnosis. The literature indicates that up to about 50% of uveitic cases have no identified causes. PMID- 19187895 TI - Enlargement of internal carotid artery aneurysm presenting with severe visual sequela: a case report and anatomy review. AB - BACKGROUND: The majority of intracranial aneurysms arise from the internal carotid artery (ICA) circulation. The proximity of cranial nerves II to VI to the pathway of the internal carotid artery make them susceptible to damage from an ICA aneurysm, which may cause a variety of neurologic effects. Although vision loss is a relatively uncommon sequela of an ICA aneurysm, compression of the optic nerve by an aneurysm can cause visual field loss, decreased visual acuity, or both. CASE REPORT: An 85-year-old man with sudden-onset, painless, and profound monocular vision loss from compression of the optic nerve caused by an enlarged internal carotid artery aneurysm is described. Clinical presentation, detection, and treatment of an ICA aneurysm are discussed. CONCLUSIONS: An intracranial aneurysm or other compromise to the cerebral circulation should be ruled out in cases of vision loss that remain unexplained after a thorough ocular health evaluation, especially in patients with a history of cardiovascular disease or head injury. PMID- 19187896 TI - Advanced visual field loss secondary to optic nerve head drusen: case report and literature review. AB - BACKGROUND: Optic nerve head drusen (ONHD) is a relatively uncommon condition that results from calcific degeneration of axons within the optic nerve. The abnormal drusen bodies can enlarge, compressing normal nerve structures, and ultimately may result in vision loss. Drusen often are discovered through clinical evaluation with a dilated funduscopic examination. Ancillary testing, including computed tomographic (CT) imaging, B-scan ultrasonography, autofluorescence imaging, nerve fiber layer imaging, and threshold visual field evaluation are helpful to confirm the existence of ONHD and to evaluate for progression of this condition. CASE REPORT: This case report discusses the clinical presentation of a patient with advanced visual field loss from ONHD and the ancillary testing used to confirm the diagnosis. A complete review of literature on ONHD is discussed. CONCLUSIONS: Currently, there is no cure or direct treatment for progressive vision loss or complications that may develop from ONHD. Useful diagnostic tools include serial automated threshold visual fields, nerve fiber layer analysis, and fundus photography. It is suggested that ocular hypotensive agents be used to lower intraocular pressure prophylactically to prevent further nerve fiber layer and optic nerve damage. PMID- 19187897 TI - Hepatitis C virus, splenic vein thrombosis, and lymphoma. PMID- 19187898 TI - Palmoplantar pustulosis and acrodermatitis in a patient treated with infliximab for Crohn's sacroiliitis. PMID- 19187899 TI - A 50-year-old woman with unexplained recurrent pancreatitis. PMID- 19187901 TI - Re: EP challenges in adult congenital heart disease. PMID- 19187903 TI - Effect of pulmonary vein isolation on the distribution of complex fractionated electrograms in humans. AB - BACKGROUND: Targeting of complex fractionated electrograms (CFEs) has been used as an adjunctive strategy to pulmonary vein isolation (PVI) in patients with persistent atrial fibrillation (AF). However, it is unclear whether CFEs should be targeted before or after PVI. OBJECTIVES: The purpose of this study was to examine the effect of PVI on CFE distribution in humans. METHODS: We compared left atrial (LA) CFE maps acquired using the NavX system before and after PVI in patients with persistent AF. CFE maps were constructed from bipolar electrograms acquired from a circular mapping catheter. At each point, the mean AF cycle length (CL) was calculated automatically by averaging the intervals between deflections over a 4-second window. Sites with mean CL < or =120 ms were considered CFE+. RESULTS: A total of 22 consecutive patients (82% male, age 58 +/ 9 years) were studied. At baseline, 47% of the LA was encompassed by electrograms with CL <120 ms. PVI had a significant effect on CFE characteristics, with an increase in mean LA AF CL (144 ms pre-PVI vs. 214 ms post-PVI; P <.01) and a decrease in CFE+ LA surface area (47% vs 23%; P <.01). There was significant reduction in CFE burden after PVI in both PV (50% vs. 6%; P <.01) and non-PV (61% vs. 39%; P <.01) regions. CONCLUSIONS: In patients with persistent AF, PVI results in a significant decrease in both PV and non-PV areas of CFE. To limit extensive LA ablation, PVI should be performed before targeting CFE when a combined approach is pursued. PMID- 19187902 TI - Systematic electrocardioversion for atrial fibrillation and role of antiarrhythmic drugs: a substudy of the SAFE-T trial. AB - BACKGROUND: Energy levels for electrocardioversion in atrial fibrillation (AF) have been empiric, and the influence of antiarrhythmic therapy compared with placebo is largely unknown. OBJECTIVE: The purpose of this study was to determine systematically the energy levels for electrocardioversion in patients with persistent AF and to define the influence of antiarrhythmic therapy. METHODS: Patients (n = 665) with persistent AF were randomized to amiodarone, sotalol, or placebo. Rate control, if necessary, was achieved with digoxin, diltiazem, or verapamil. Among the 665 patients, 504 who did not achieve sinus rhythm at day 28 had electrocardioversion systematically by a prespecified four-step protocol as follows: monophasic shocks-100, 200, 360, 360 J; or biphasic shocks-150, 175, 200, 200 J sequentially. Energy levels and shock waveforms (monophasic/biphasic) for successful electrocardioversion (sinus rhythm for at least 1 minute) and use of antiarrhythmic therapy and calcium channel blockers were recorded. RESULTS: Electrocardioversion was successful in 371 (71.6%) of 504 patients: 72%, 73.5%, and 67.9% for patients assigned to amiodarone, sotalol, and placebo, respectively. Overall, after adjustments for age, body mass index (BMI), history of AF, shock waveforms, left atrial size, and ejection fraction, both amiodarone (odds ratio [OR]: 2.16, 95% confidence interval [CI]: 1.24-3.77, P <.01) and sotalol (OR: 1.92, 95% CI: 1.11-3.33, P = .02) significantly facilitated successful electrocardioversion compared with placebo. Calcium channel blockers had no effect on the success rate of electrocardioversion. Success of electrocardioversion was associated with lower BMI, AF history < or =1 year, and older age. Compared with placebo, patients taking amiodarone were significantly more likely to achieve successful electrocardioversion in step 1 (OR: 2.73, 95% CI: 1.11-6.74, P = .03) and step 3 (OR: 1.86, 95% CI: 1.00-3.44, P = .05) but not in steps 2 and 4. Sotalol was superior to placebo in step 4 (OR: 2.58, 95% CI: 1.02-6.52, P = .05) and trended in step 2 (OR: 1.7, 95% CI: 0.98-3.07, P = .06). Successful electrocardioversion was seen in 11%, 29%, 38%, and 29% in steps 1, 2, 3, and 4, respectively. Compared with monophasic shocks, biphasic shocks achieved higher success rates for step 1 (P <.001) and step 2 (P <.01), respectively. Antiarrhythmic therapy did not influence the total number of energy steps used for the patients with successful electrocardioversion. However, biphasic shocks, lower BMI, and AF duration < or =1 year were associated with less energy step used for successful cardioversion. CONCLUSION: Amiodarone and sotalol facilitated successful electrocardioversion, which could be achieved in a stepwise fashion. Upon achievement of successful electrocardioversion, amiodarone is superior to placebo, and sotalol has a lesser effect. Antiarrhythmic drugs had no effect on the total number of energy step use in patients who had successful electrocardioversion. Calcium channel blockers had no influence on the success rate in achieving sinus rhythm. Successful electrocardioversion was associated with lower BMI and AF history < or =1 year. Lower energy use was associated with biphasic shocks, lower BMI, and AF duration < or =1 year. PMID- 19187904 TI - Temporal left atrial lesion formation after ablation of atrial fibrillation. AB - BACKGROUND: Atrial fibrillation (AF) ablation uses radiofrequency (RF) energy to induce thermal damage to the left atrium (LA) in an attempt to isolate AF circuits. This injury can be seen using delayed enhancement magnetic resonance imaging (DE-MRI). OBJECTIVE: The purpose of this study was to describe DE-MRI findings of the LA in the acute and chronic stages postablation. METHODS: Twenty five patients were scanned at two time points postablation. The first group (n = 10) underwent DE-MRI at 24 hours and at 3 months. The second group (n = 16) was scanned at 3 months and at 6 or 9 months. One patient had three scans (24 hours, 3 months, 9 months) and was included in both groups. The location and extent of enhancement were then analyzed between both groups. RESULTS: The median change in LA wall injury between 24 hours and 3 months was -6.38% (range -11.7% to 12.58%). The median change in LA wall injury between 3 months and later follow-up was +2.0% (range -4.0% to 6.58%). There appears to be little relationship between the enhancement at 24 hours and 3 months (R(2) = 0.004). In contrast, a strong correlation is seen at 3 months and later follow-up (R(2) = 0.966). Qualitative comparison revealed a stronger qualitative relationship between MRI findings at 3 months and later follow-up than at 24 hours and 3 months. CONCLUSION: RF-induced scar appears to have formed by 3 months postablation. At 24 hours postablation, DE-MRI enhancement appears consistent with a transient inflammatory response rather than stable LA scar formation. PMID- 19187905 TI - Risk of ischemic stroke after new-onset atrial fibrillation in patients with hyperthyroidism. AB - BACKGROUND: Hyperthyroidism is one of the most common reversible causes of atrial fibrillation (AF); nevertheless, the risk of ischemic stroke in patients with hyperthyroidism who present with new-onset AF is unclear. OBJECTIVE: This study sought to investigate the clinical outcome of hyperthyroidism-induced AF with regard to risk of ischemic stroke risk. METHODS: We prospectively studied the incidence, time course, and clinical predictors for ischemic stroke in patients with hyperthyroidism-induced AF (n = 160). They were compared with age- and sex matched cohorts of hyperthyroid patients without AF (n = 160) and AF patients without hyperthyroidism (n = 160). RESULTS: Baseline characteristics were comparable among the 3 groups. At 1 year, 86 hyperthyroid patients with AF (54%) and 92 patients with nonthyroid AF (58%) had spontaneous or pharmacological sinus conversion (P = .20). Ischemic stroke was observed in 15 hyperthyroid patients with AF (9.4%) versus 5 patients with nonthyroid AF (3.1%, P = .02), and 1 hyperthyroid patient without AF (0.6%, P < .001). Furthermore, the majority of ischemic stroke (>70%) in patients with AF occurred within the first 30 days of presentation, whereas AF was still present. Cox regression analysis showed that hyperthyroidism (hazard ratio [HR]: 3.5, 95% confidence interval [CI]: 1.15 to 10.42, P = .03) and persistent AF (HR: 13.0, 95% CI: 2.88 to 58.80, P < .01) predicted the occurrence of ischemic stroke; warfarin therapy reduced the risk of ischemic stroke (HR: 0.17, 95% CI: 0.04 to 0.79, P = .02). CONCLUSION: In hyperthyroid patients who presented with new-onset AF, there was an increased risk of ischemic stroke clustering during the initial phase of presentation. This should prompt early use of anticoagulation therapy in hyperthyroid patients with AF. PMID- 19187906 TI - Catheter ablation of atrial fibrillation via superior approach in patients with interruption of the inferior vena cava. AB - BACKGROUND: Percutaneous transcatheter ablation of atrial fibrillation (AF) in patients with interruption of inferior vena cava (IVC) has not been reported in the literature. OBJECTIVE: The purpose of this article was to demonstrate the safety and feasibility of the superior approach via the right internal jugular vein in performing catheter ablation of AF. METHODS: We performed AF ablation in 3 patients (mean age: 51.7 +/- 18.5 years, 2 paroxysmal AF and 1 persistent AF) with complete interruption of IVC. Transseptal puncture was performed via the right internal jugular vein with a long sheath and manually curved Brockenbrough needle to facilitate the tip downward to the FO. Three-dimensional (3D) mapping was performed in 2 patients. Electrical isolation of each pulmonary vein (PV) was confirmed by a circular mapping catheter. Bidirectional block at the RA isthmus was achieved in 1 patient with clinically documented typical atrial flutter. RESULTS: In all patients, AF ablation after transseptal puncture via the superior approach was successfully performed without complications. Selective PV isolation of arrhythmogenic PV was done in 1 patient, and circumferential bilateral antral ablations were done in 2 patients. In 1 patient with persistent AF, linear ablations of left atrial roof and perimitral and RA isthmus were done after electrical isolation of all PVs. At a mean follow-up of 18.7 +/- 15.5 months, arrhythmias were free without any antiarrhythmic drugs in all patients. CONCLUSION: AF ablation via the superior approach is a safe and feasible alternative technique when a femoral venous approach is not available. PMID- 19187907 TI - Gender differences and risk of ventricular tachycardia or ventricular fibrillation. AB - BACKGROUND: Healthy women have longer QT intervals and more drug-induced proarrhythmia compared to men, yet those given implantable cardioverter difibrillators (ICDs) for ischemic cardiomyopathy have fewer episodes of ventricular tachycardia/ventricular fibrillation (VT/VF) than men. The role of repolarization duration and stability in arrhythmogenesis in men and women with structural heart disease has not been explored. OBJECTIVES: The purpose of this study was to analyze repolarization differences between men and women and their relation to the risk of VT/VF. METHODS: Multicenter Automatic Defibrillator Trial II study patients underwent 10-minute, resting digitized recordings at study entry. QT and heart rate were measured for each beat with a semiautomated method. QT variance was normalized for mean QT (QTVN) or for heart rate variance (QTVI). Spectral analysis of heart rate and QT time series was performed; coherence was indexed to quantify consistency of heart rate and QT power spectra. The incidence of VT/VF was determined by ICD interrogation. RESULTS: There were 805 usable recordings (142 females); 463 received ICDs (86 females). There was no gender difference in mean or median QT, QTc, or heart rate. QTVN and QTVI were slightly (but significantly) higher, and the mean coherence was lower in women. In a Cox multivariate analysis, increased QTVN or QTVI (top quartile) was associated with a significantly higher risk for VT/VF in men (QTVN hazard ratio (HR) 2.2; confidence interval [CI] 1.4-3.4; P = .001; QTVI HR 1.9; CI 1.2-3.0; P = .006) but not in women, while reduced coherence (bottom quartile) predicted VT/VF in women (HR 3.3; CI 1.2-9.0; P = .021) but not in men. CONCLUSIONS: In post myocardial infarcation patients with depressed ejection fraction, both women and men manifest increased temporal variability in the QT interval. In men, QT variability by itself raised arrhythmic risk. In women, however, QT variability dissociated from HR variability (low coherence) appeared to be a uniquely significant predictor of arrhythmic events. PMID- 19187908 TI - Sex discrimination and the QT interval. PMID- 19187910 TI - Radiofrequency ablation for ventricular tachycardia in patients with cardiac sarcoidosis: is it worth trying? PMID- 19187909 TI - Role of radiofrequency catheter ablation of ventricular tachycardia in cardiac sarcoidosis: report from a multicenter registry. AB - BACKGROUND: Management of ventricular tachycardia (VT) is challenging in patients with cardiac sarcoidosis. OBJECTIVE: The purpose of this study was to assess the response of VT in patients with cardiac sarcoidosis to medical therapy and radiofrequency ablation. METHODS: Forty-two patients with a diagnosis of cardiac sarcoidosis based on the Japanese Health Ministry criteria were followed. When VT occurred, a stepwise approach was used: implantable cardioverter-defibrillator placement, immunosuppressive agents, antiarrhythmic medications, then radiofrequency ablation. RESULTS: In nine patients (age 46.7 +/- 8.6 years; ejection fraction 42 +/- 14%), VT was not controlled by medical therapy, and radiofrequency ablation was performed. A total of 44 VTs (mean cycle length 348 +/- 78 ms) were induced. Endocardial radiofrequency ablation was performed in eight patients (right ventricular in 5, left ventricular in 3) and epicardial radiofrequency ablation in one patient. In 4 of 5 patients with right ventricular VTs, a peritricuspid circuit was identified. Critical areas were identified for 21 (48%) of 44 VTs, resulting in elimination of 31 (70%) of 44 VTs. The most frequent VT circuit was reentry in the peritricuspid area. This type of VT was eliminated in all patients. Arrhythmic events decreased from 271 +/- 363 episodes preablation to 4.0 +/- 9.7 postablation. All patients had either a decrease (n = 4) or complete elimination (n = 5) of VT during mean follow-up of 19.8 +/- 19.6 months. CONCLUSION: Catheter ablation of VT in patients with cardiac sarcoidosis refractory to medical therapy is effective in eliminating VT or markedly reducing the VT burden. The disease process in cardiac sarcoidosis often involves a specific area in the basal right ventricle predisposing to peritricuspid reentry. PMID- 19187911 TI - Electrophysiologic characteristics and catheter ablation of focal atrial tachycardia with more than one focus. AB - BACKGROUND: Information about the electrophysiologic characteristics and long term outcome of catheter ablation in patients with multiple focal atrial tachycardia (AT) is limited. OBJECTIVE: The purpose of this study was to investigate the electrophysiologic characteristics and long-term outcome of catheter ablation in patients with multiple focal AT. METHODS: Two hundred fifty one patients who were referred for radiofrequency catheter ablation of focal AT were included for analysis. RESULTS: Forty-four patients who had focal AT with more than one focus were identified. Comparing focal AT with a single focus to that with more than one focus, the existence of a left atrial focus, cardiovascular comorbidity, nonparoxysmal tachycardia, shortest tachycardia cycle length, success rate of the ablation, and procedure time all differed. Multivariate logistic analysis revealed that a left atrial focus, cardiovascular comorbidity, and shortest tachycardia cycle length were independent predictors of focal AT with more than one focus. Noncontact mapping of the right atrium revealed larger low-voltage zone and longer total activation time for focal AT with more than one focus. Patients who had focal AT with more than one focus and a failed ablation had a greater number of focal ATs and mechanisms of nonparoxysmal tachycardia. Multivariate logistic analysis revealed that only the number of focal ATs predicted a failed ablation. CONCLUSION: Focal ATs with more than one focus have different electrophysiologic characteristics. This study provides new insight into the development and atrial remodeling of focal AT with multiple foci. PMID- 19187912 TI - Bad lead? Good lead? What should we believe? PMID- 19187913 TI - A KCNH2 branch point mutation causing aberrant splicing contributes to an explanation of genotype-negative long QT syndrome. AB - BACKGROUND: Genetic screening of long QT syndrome (LQTS) fails to identify disease-causing mutations in about 30% of patients. So far, molecular screening has focused mainly on coding sequence mutations or on substitutions at canonical splice sites. OBJECTIVE: The purpose of this study was to explore the possibility that intronic variants not at canonical splice sites might affect splicing regulatory elements, lead to aberrant transcripts, and cause LQTS. METHOD: Molecular screening was performed through DHPLC and sequence analysis. The role of the intronic mutation identified was assessed with a hybrid minigene splicing assay. RESULTS: A three-generation LQTS family was investigated. Molecular screening failed to identify an obvious disease-causing mutation in the coding sequences of the major LQTS genes but revealed an intronic A-to-G substitution in KCNH2 (IVS9-28A/G) cosegregating with the clinical phenotype in family members. In vitro analysis proved that the mutation disrupts the acceptor splice site definition by affecting the branch point (BP) sequence and promoting intron retention. We further demonstrated a tight functional relationship between the BP and the polypyrimidine tract, whose weakness is responsible for the pathological effect of the IVS9-28A/G mutation. CONCLUSIONS: We identified a novel BP mutation in KCNH2 that disrupts the intron 9 acceptor splice site definition and causes LQT2. The present finding demonstrates that intronic mutations affecting pre-mRNA processing may contribute to the failure of traditional molecular screening in identifying disease-causing mutations in LQTS subjects and offers a rationale strategy for the reduction of genotype-negative cases. PMID- 19187914 TI - Intronic variants and splicing errors in cardiovascular diseases. PMID- 19187915 TI - Nerve supply of the human pulmonary veins: an anatomical study. AB - BACKGROUND: Atrial ectopic discharges originating in the pulmonary veins (PVs) are known to initiate atrial fibrillation (AF), which may be terminated by catheter-based PV isolation. Because a functional relationship exists between cardiac autonomic effects and PVs in arrhythmogenesis, it has been suggested that discharges of the nerves that proceed to the PVs and interconnect with intrinsic ganglionated nerve plexuses are potential triggers of AF in man. OBJECTIVE: This study sought to determine the characteristics and distribution of neural routes by which autonomic nerves supply the human PVs. METHODS: We examined the intrinsic neural structures of 35 intact (nonsectioned) left atrial (LA)-PV complexes stained transmurally for acetylcholinesterase using a stereomicroscope. RESULTS: The epicardial ganglionated nerves pass onto the extrapulmonary segments of the human PVs from the middle, left dorsal, and dorsal right atrial subplexuses. The left and right inferior PVs involved a lesser number of ganglia than the left and right superior PVs. Abundant extensions of epicardial nerves penetrate transmurally the PV walls and form a patchy neural network beneath the endothelium of PVs. The subendothelial neural meshwork with numerous free nerve endings, which appeared to be typical sensory compact nerve endings, was mostly situated at the roots of the 4 PVs. No ganglia were identified beneath the endothelium of the human PVs. CONCLUSION: The richest areas containing epicardial ganglia, from which intrinsic nerves extend to the human PVs, are concentrated at the inferior surface of both the inferior and left superior PVs. Therefore, these locations might be considered as potential targets for focal pulmonary vein ablation in catheter-based therapy of AF. PMID- 19187916 TI - Innervation of the human pulmonary veins. PMID- 19187917 TI - Segmentation and registration of three-dimensional rotational angiogram on live fluoroscopy to guide atrial fibrillation ablation: a new online imaging tool. AB - BACKGROUND: Three-dimensional rotational atriography (3DATG) was developed to supplement two-dimensional fluoroscopy with 3D volume reconstruction of the left atrium (LA), pulmonary veins (PV), and other structures. Until recently, 3DATG images could only be viewed separately and were not suitable to directly guide atrial fibrillation (AF) ablation. OBJECTIVE: The purpose of this study was to evaluate the feasibility and accuracy of intraprocedural 3DATG. METHODS: Three dimensional rotational atriography with right atrial contrast injection was obtained using a Philips Allura Xper FD10 system in 30 patients with symptomatic AF who also underwent preprocedural computed tomographic (CT) scan. RESULTS: The majority (93%) of 3DATG image reconstructions were useful for guidance of catheter ablation. Nearly all PVs (94%), LA appendage (89%), and esophagus (100%) were successfully segmented. Measured PV ostial diameters compared using 3DATG and CT showed close concordance. Registration and re-registration of 3DATG overlay image was easily achieved with thoracic landmarks and validated by catheter placement demonstrating minimal discrepancy. Endoscopic views allowed for improved visualization of ostial position, dimensions, and navigation within the antrum. Lesion tagging on 3DATG overlay enhanced ablation guidance. Radiation exposure with 3DATG was significantly reduced compared with preprocedural CT scan (2.1 +/- 0.3 mSv vs 13.8 +/- 2.4 mSv, P <.001). CONCLUSION: Intraprocedural 3DATG imaging during AF ablation with online segmentation and superimposition on live fluoroscopy is feasible. Overlay provides valuable and accurate information on 3D surface outline and endoscopic PV location. Three-dimensional rotational atriography overlay is a new imaging method with reduced radiation exposure that may replace preprocedural CT scan for catheter navigation and ablation of AF. PMID- 19187918 TI - Predicting the cause of syncope from clinical history in patients undergoing prolonged monitoring. AB - BACKGROUND: Syncope may be the result of primary bradycardia or tachycardia, vasovagal syncope, or noncardiac syncope. Risk factors and outcome scores to predict prognosis in patients with syncope have been developed. Although these correlate with morbidity and mortality in patients with syncope, their relationship with the mechanism of syncope has not been investigated. OBJECTIVE: The purpose of this study was to identify clinical predictors of primary bradycardia in a cohort of patients undergoing prolonged monitoring for unexplained syncope. METHODS: One hundred nineteen patients underwent prolonged monitoring with an implantable or external loop recorder after assessment at a single-center, tertiary care arrhythmia service. Fifty-two patients with recurrent syncope during monitoring were classified according to the mechanism of syncope (International Study on Syncope of Uncertain Etiology [ISSUE] classification). Clinical predictors of primary arrhythmic syncope were identified. RESULTS: Twenty patients were classified with primary arrhythmia and 32 patients were classified with nonarrhythmic syncope. Five clinical variables were associated with primary arrhythmia: left bundle branch block, structural heart disease, and syncope without prodrome increased the likelihood of primary arrhythmia; a normal baseline ECG and history of syncope in childhood decreased the likelihood of primary arrhythmia. After multiple logistic regression, risk factors for the diagnosis of primary arrhythmia included syncope without warning symptoms and structural heart disease. The presence of left bundle branch block correlated perfectly with primary arrhythmia, whereas a normal ECG reduced the likelihood of primary arrhythmia. CONCLUSION: Clinical predictors of primary arrhythmia in patients with recurrent syncope include normal ECG and structural heart disease. Left bundle branch block is an important finding in patients with unexplained syncope. PMID- 19187919 TI - Relationship between right cervical vagus nerve stimulation and atrial fibrillation inducibility: therapeutic intensities do not increase arrhythmogenesis. AB - BACKGROUND: Strong vagus nerve stimulation (VNS) is routinely used to induce and maintain atrial fibrillation (AF) in acute animal studies. Taken as a surrogate of increased vagal tone, such observations suggest an arrhythmogenic role of VNS in AF. In contrast, VNS has been demonstrated to have profound therapeutic effects in heart failure and other ailments. OBJECTIVE: The purpose of this study was to examine the relationship between right cervical VNS and AF, especially the potential arrhythmogenic effects of therapeutic VNS. METHODS: The relationship between VNS intensities and AF inducibility was studied in eight acute dogs at baseline and four different levels of VNS, which were set to prolong spontaneous sinus cycle length (SCL) by 20%, 40%, 60%, or 100%. The effect of mild VNS treatment on AF induction was further investigated in six chronically instrumented conscious dogs. These dogs were implanted with right cervical VNS stimulators and specialized atrial pacemakers. VNS intensity was titrated to slow the sinus rate by 10%. RESULTS: In acute studies, it was found that mild to moderate VNS (i.e., producing < or =40% SCL prolongation) did not increase AF inducibility, while strong VNS (i.e., producing > or =60% SCL prolongation) did. In chronic studies, compared with controls, AF induction did not change during the 4-week VNS treatment. CONCLUSIONS: AF inducibility by right cervical VNS is intensity dependent: strong VNS (producing > or =60% SCL prolongation) facilitates AF, while moderate VNS (producing < or =40% SCL prolongation) appears not to affect AF. The nonarrhythmogenic effect of therapeutic chronic VNS was further verified in conscious animals. We conclude that VNS with moderate intensities can be used to deliver therapeutic benefits without arrhythmogenic risk. PMID- 19187920 TI - Heart failure enhances susceptibility to arrhythmogenic cardiac alternans. AB - BACKGROUND: Although heart failure (HF) is closely associated with susceptibility to sudden cardiac death (SCD), the mechanisms linking contractile dysfunction to cardiac electrical instability are poorly understood. Cardiac alternans has also been closely associated with SCD, and has been linked to a mechanism for amplifying electrical heterogeneities in the heart. However, previous studies have focused on alternans in normal rather than failing myocardium. OBJECTIVE: This study sought to investigate the hypothesis that HF enhances susceptibility to arrhythmogenic cardiac alternans. METHODS: High-resolution transmural optical mapping was performed in canine wedge preparations from normal (n = 8) and HF (n = 8) hearts produced by rapid ventricular pacing. RESULTS: HF significantly (P < .004) lowered the heart rate (HR) threshold for action potential duration alternans (APD-ALT) from 236 +/- 25 beats/min to 185 +/- 25 beats/min. In dual optical mapping of action potentials and intracellular Ca experiments (n = 16), HF lowered the HR threshold for Ca-ALT (beat-to-beat alternations of cellular Ca cycling) from 238 +/- 35 to 177 +/- 26 beats/min (P < .005). Importantly: (1) Ca ALT always either developed at slower HR or simultaneously with APD-ALT in the same cells, and (2) the magnitude of Ca-ALT and APD-ALT were closely correlated (P < .05). HF similarly lowered the HR threshold for Ca-ALT in isolated myocytes under nonalternating action potential clamp, indicating that HF enhances susceptibility to cellular alternans independent of HF-associated changes in repolarization. Importantly, HF significantly (P < .02) lowered the HR threshold for spatially discordant arrhythmogenic alternans (different regions of cells alternating in opposite phase, DIS-ALT). Ventricular fibrillation (VF) was induced in 88% of HF preparations, but only 12% of normal preparations (P < .003) and was uniformly preceded by development of DIS-ALT. CONCLUSION: Heart failure increases the susceptibility to arrhythmogenic cardiac alternans, which arises from HF-induced impairment in calcium cycling. PMID- 19187921 TI - From nonuniform repolarization to abnormal calcium dynamics: the elusive culprit for ventricular arrhythmias in heart failure. PMID- 19187922 TI - Isolated focus of ventricular fibrillation. AB - Mechanisms underlying the initiation of ventricular tachycardia have been described as either rapidly firing focus initiated by triggered activity or automaticity, or a reentrant wavefront initiated by a premature ventricular depolarization. Ventricular fibrillation can occur after ventricular tachycardia; however, uncertainty exists concerning its mechanism, whether it is caused by multiple wavelets, mother rotor, or a combination of both. This report describes the finding of a spontaneously isolated focus of ultrarapid ventricular activity recorded and ablated in a patient with nonischemic cardiomyopathy and ventricular tachycardia. PMID- 19187923 TI - Differences in twelve-lead electrocardiogram between symptomatic and asymptomatic subjects with short QT interval. PMID- 19187924 TI - Unusual response to a premature ventricular complex introduced during an episode of paroxysmal supraventricular tachycardia: what is the mechanism? PMID- 19187925 TI - Failure of a dual-chamber implantable-cardioverter-defibrillator to pace: malfunction or normal function? PMID- 19187926 TI - Catheter ablation in transposition of the great arteries with Mustard or Senning baffles. PMID- 19187928 TI - Recent developments in the preparation and regeneration of activated carbons by microwaves. AB - To date, microwave energy has been widely developed and applied to almost every field of chemistry. In many cases, microwave technology has proven to remarkably reducing costs, accelerating reaction rates, improving yields and selectively activating. This paper presents a state of art review of microwave technology, its background studies, fundamental chemistry and industrial applications. With the renaissance of activated carbon, there has been a steadily growing interest in this research field. The review provides a summary on recent development in preparation and regeneration of activated carbons. The key advance of introducing microwave energy has been highlighted relative to conventional methods. Moreover, the major drawbacks, challenges with its future expectation are presented and discussed. Conclusively, microwave energy is predicted to be a potentially viable and powerful replacement for fuel technology in various areas, while its progress represents an expanding field in the area of adsorption science. PMID- 19187927 TI - Epicardial ablation of left ventricular outflow tract tachycardia via the coronary sinus. PMID- 19187930 TI - The first total synthesis of 7-O-beta-D-glucopyranosyl-4'-O-alpha-L rhamnopyranosyl apigenin via a hexanoyl ester-based protection strategy. AB - The first total synthesis of 7-O-beta-D-glucopyranosyl-4'-O-alpha-L rhamnopyranosyl apigenin 1, which exhibits good anti-hepatitis B virus and anti stroke activities, was accomplished in six steps and 20% overall yield from apigenin. Another synthetic route, in which the target was obtained in seven steps, was also developed to prove the utility of a hexanoyl ester-based orthogonal protection strategy. The hexanoyl protection strategy provided all the flavonoid intermediates with good solubility and reactivity, enabled efficient selective protection and glycosylation, and provided a practical and effective synthetic strategy for flavonoids, starting from commercially available flavone. PMID- 19187929 TI - The effects of shoe traction and obstacle height on lower extremity coordination dynamics during walking. AB - This study aims to investigate the effects of shoe traction and obstacle height on lower extremity relative phase dynamics (analysis of intralimb coordination) during walking to better understand the mechanisms employed to avoid slippage following obstacle clearance. Ten participants walked at a self-selected pace during eight conditions: four obstacle heights (0%, 10%, 20%, and 40% of limb length) while wearing two pairs of shoes (low and high traction). A coordination analysis was used and phasing relationships between lower extremity segments were examined. The results demonstrated that significant behavioral changes were elicited under varied obstacle heights and frictional conditions. Both decreasing shoe traction and increasing obstacle height resulted in a more in-phase relationship between the interacting lower limb segments. The higher the obstacle and the lower the shoe traction, the more unstable the system became. These changes in phasing relationship and variability are indicators of alterations in coordinative behavior, which if pushed further may have lead to falling. PMID- 19187931 TI - The structure of the polysaccharide O-chain of the LPS from Acinetobacter baumannii strain ATCC 17961. AB - The gram-negative bacterium Acinetobacter baumannii strain ATCC17961 has been used by several laboratories in mouse models of respiratory A. baumannii infection, and a study of the role of its lipopolysaccharide in the pathogenicity is of interest. The structure of the O-deacylated polysaccharide O-chain component of its LPS has been determined by 2D NMR spectroscopy and mass spectrometry methods, and by the structural identification of oligosaccharides obtained by sequential application of the Smith degradation of the O-antigen. The O-chain was determined to be a polymer of a branched pentasaccharide repeating unit composed of 2,3-diacetamido-2,3-dideoxy-D-glucuronic acid, 2-acetamido-2 deoxy-D-glucose, 2-acetamido-2-deoxy-D-galactose, D-glucose, and D-galactose, and has the following structure: [carbohydrate sequence see in text]. PMID- 19187932 TI - Pelvic ring fracture--dislocation in a 2400-year-old Etruscan man. PMID- 19187933 TI - A 4-year review of human bite injuries presenting to emergency medicine and proposed evidence-based guidelines. AB - Human bite injuries are a common injury, roughly accounting for around 0.1% of attendances to Emergency Medicine departments. Morbidities associated with such injuries are legion. Of paramount importance is the potential for infection of the wound site, both bacterial and viral. A retrospective 4-year review of 3136 case notes was conducted, identifying 421 human bites. This amounts to one every 3 days! The majority of those bitten were young males (male:female ratio=3:1; 44% comprising the age group 16-25 years). The management of these wounds was found to be poor. 17% of patients did not receive any antibiotic cover; 21% of patients either did not have tetanus prophylaxis administered when required or had a tetanus booster when they were already covered; 34% of patients either did not receive a hepatitis B booster when one was required or received one when they were already covered. This lack of effective documentation, along with errors in addressing prophylaxis of the infective agents, may have profound medico-legal consequences. The author believes that the findings will be reproduced in other centres and in view of the inadequacies highlighted by this work, it is necessary to introduce an evidence-based protocol for the comprehensive management of the human bite. Using United Kingdom Department of Health evidence-based guidelines, the author proposes such a pathway. PMID- 19187934 TI - The research evidence published in high impact nursing journals between 2000 and 2006: a quantitative content analysis. AB - BACKGROUND: Evidence-based practice has become an imperative for efficient, effective and safe practice. Furthermore, evidences emerging from published research are considered as valid knowledge sources to guiding practice. OBJECTIVES: The aim of this paper is to review all research articles published in the top 10 general nursing journals for the years 2000-2006 to identify the methodologies used, the types of evidence these studies produced and the issues upon which they endeavored. DESIGN: Quantitative content analysis was implemented to study all published research papers of the top 10 general nursing journals for the years 2000-2006. METHODS: The top 10 general nursing journals were included in the study. The abstracts of all research articles were analysed with regards the methodologies of enquiry, the types of evidence produced and the issues of study they endeavored upon. Percentages were developed as to enable conclusions to be drawn. RESULTS: The results for the category methodologies used were 7% experimental, 6% quasi-experimental, 39% non-experimental, 2% ethnographical studies, 7% phenomenological, 4% grounded theory, 1% action research, 1% case study, 15% unspecified, 5.5% other, 0.5% meta-synthesis, 2% meta-analysis, 5% literature reviews and 3% secondary analysis. For the category types of evidence were 4% hypothesis/theory testing, 11% evaluative, 5% comparative, 2% correlational, 46% descriptive, 5% interpretative and 27% exploratory. For the category issues of study were 45% practice/clinical, 8% educational, 11% professional, 3% spiritual/ethical/metaphysical, 26% health promotion and 7% managerial/policy. CONCLUSIONS: Published studies can provide adequate evidences for practice if nursing journals conceptualise evidence emerging from non experimental and qualitative studies as relevant types of evidences for practice and develop appropriate mechanisms for assessing their validity. Also, nursing journals need to increase and encourage the publication of studies that implement RCT methodology, systematic reviews, meta-synthesis and meta-analysis methodologies. Finally, nursing journals need to encourage more high quality research evidence that derive from interpretative, theory testing and evaluative types of studies that are practice relevant. PMID- 19187935 TI - Intracellular labile iron promotes firm adhesion of human monocytes to endothelium under flow and transendothelial migration: Iron and monocyte endothelial cell interactions. AB - Monocyte infiltration across the endothelium is part of the innate immune response, however it may contribute to severity of chronic conditions. We have investigated the effects of iron on the cytokine-mediated recruitment of monocytes to the endothelium, using a physiological flow model and a monocyte transendothelial migration model. Under flow, iron loading to endothelial cells promoted an increased number of tumor necrosis factor-alpha-mediated firm arrest of human monocytes. Similarly, an increased number of firmly adhered monocytes were observed in conditions in which monocytes were iron-loaded, compared to the non-iron-loaded conditions. In both iron-loaded and non-iron-loaded conditions, blockade of the alpha4 and beta2 integrins restored similar number and velocity of monocyte rolling, suggesting that iron did not modulate rolling interactions. However, with the integrin blockade, the number of firmly adhered cells remained higher in iron-loaded conditions than in control conditions, suggesting that iron could have modulated receptors other than the blocked integrins to promote firm arrest. Iron loading indeed upregulated expression of chemokine receptors, CC receptor-2 and CXC receptor-2, but not platelet endothelial cell adhesion molecule-1. This effect concomitantly promoted monocyte chemotactic protein-1 dependent transendothelial migration. In addition, iron-induced firm adhesion and transmigration were counteracted by iron chelation. These data reveal an immunomodulatory function of iron in the cascade of events of cytokine-mediated monocyte infiltration across endothelium, and therefore suggests the role for iron in inflammatory conditions underlying diseases like atherosclerosis and neurodegeneration. PMID- 19187936 TI - Regulation and possible role of endocannabinoids and related mediators in hypercholesterolemic mice with atherosclerosis. AB - In this study we analysed the possible modulation of endocannabinoids and related molecules during atherosclerosis development in mice. Wild-type and apolipoprotein E knockout (ApoE(-/-)) mice were fed either normal chow or high cholesterol diet for 8-12 weeks, and tissue endocannabinoid levels were measured by liquid chromatography-mass spectrometry. We found increased levels of 2-AG in aortas and visceral adipose tissue (VAT) of ApoE(-/-) mice fed on high cholesterol diet for 12 weeks as compared to ApoE(-/-) mice fed on normal chow or wild-type mice fed on cholesterol. No significant difference in 2-AG levels was observed after 8 weeks of diet, and no changes in anandamide levels were found in any group. The levels of the anandamide-related mediators with anti-inflammatory or anti-lipogenic properties, palmitoylethanolamide (PEA) and oleoylethanolamide (OEA), decreased or increased only in VAT or in both tissues, respectively. Endocannabinoid- and OEA/PEA-degrading enzymes were expressed by macrophages within atherosclerotic lesions. In vitro, 2-AG and OEA-induced monocyte migration at 0.3-1microM, which corresponds to the levels observed in aortas. PEA 1microM also induced monocyte migration but counteracted the effect of 2-AG, whereas OEA enhanced it. Enhanced 2-AG levels in advanced atherosclerotic lesions may trigger the inflammatory process by recruiting more inflammatory cells and inducing extracellular matrix degradation via CB(2) receptors, and this possibility was supported in vitro but not in vivo by experiments with the CB(2) antagonist, SR144528. PMID- 19187937 TI - Suppressing the charged coupled device noise in univariate thin-layer videoscans: a comparison of several algorithms. AB - The digital processing of chromatographic thin-layer plate images has increasing popularity among last years. When using a camera instead of flatbed scanner, the charged coupled device (CCD) noise is a well-known problem-especially when scanning dark plates with weakly fluorescing spots. Various techniques are proposed to denoise (smooth) univariate signals in chemometric processing, but the choice could be difficult. In the current paper the classical filters (Savitzky-Golay, adaptive degree polynomial filter, Fourier denoising, Butterworth and Chebyshev infinite impulse response filters) were compared with the wavelet shrinkage (31 mother wavelets, 3 thresholding techniques and 8 decomposition levels). The signal obtained from 256 averaged videoscans was treated as the reference signal (with noise naturally suppressed, which was found to be almost white one). The best choice for denoising was the Haar mother wavelet with soft denoising and any decomposition level larger than 1. Satisfying similarity to reference signal was also observed in the case of Butterworth filter, Savitzky-Golay smoothing, ADPF filter, Fourier denoising and soft thresholded wavelet shrinkage with any mother wavelet and middle to high decomposition level. The Chebyshev filters, Whittaker smoother and wavelet shrinkage with hard thresholding were found to be less efficient. The results obtained can be used as general recommendations for univariate denoising of such signals. PMID- 19187938 TI - A new approach to the determination of column overload characteristics in reversed-phase liquid chromatography. AB - Column overloading is very common during the separations of basic analytes in analytical scale reversed-phase liquid chromatography (RPLC). Due to the complex interactions of ionic analytes with stationary and mobile phases, only a very small amount of ionized sample compared to the amount of nonpolar solute can be injected before the peak shape is distorted by non-linear chromatographic processes. Often the amount that can be injected before overload is observed is so small that the signal is quite noisy, thereby making the measured plate count imprecise and possibly inaccurate. The purpose of the present study was to develop a practical method for the precise measurement of the plate count and a column overload parameter using a simple but mathematically rigorous model of Langmuirian non-linear chromatography. An "overload profile", i.e. a plot of apparent plate count versus amount injected, is characterized by two parameters: the limiting plate count (N(0)) and the column sample loading capacity (omega(0.5)). The limiting plate count is the plate count that should be observed when the amount of sample injected is so small that a linear isotherm pertains. The column sample loading capacity, which is taken as the sample load that leads to a plate count equal to half of the limiting plate count, is a measure of the maximum amount of sample that can be injected into that column. The approach was tested by applying it to the study of cationic analytes in RPLC. We show that N(0) under constant conditions (column length, flow rate, mobile phase composition, etc.) is almost independent of column type (manufacturer); however, different column types (at the same length, diameter and flow rate) exhibit clear differences in their sample loading capacity (omega(0.5)). We believe that for most well packed type B columns, the column sample loading capacity and not the limiting plate count is the more important property that accounts for most of the apparent differences in peak width when different types of columns are examined. PMID- 19187939 TI - Detection of urinary markers for thiazide diuretics after oral administration of hydrochlorothiazide and altizide-relevance to doping control analysis. AB - In sports, thiazide diuretics are used to flush out previously taken prohibited substances with forced diuresis and in sports where weight classes are involved to achieve acute weight loss. Thiazide diuretics include compounds which are very unstable and hydrolyse in aqueous media. Because information regarding the urinary detection of the hydrolysis products is limited, urinary excretion profiles for the hydrolysis product 4-amino-6-chloro-1,3-benzenedisulphonamide were established in 6 healthy volunteers after oral administration of altizide (15 mg per tablet) and hydrochlorothiazide (25mg per tablet). Additionally, the excretion profile of chlorothiazide, a metabolite of altizide and hydrochlorothiazide, was also determined. A quantitative liquid-chromatographic tandem mass spectrometric method to detect the 4 substances was developed and validated. The result of this work shows that altizide is eliminated within 48 h in urine whereas hydrochlorothiazide was detectable after 120 h. Chlorothiazide was determined to be a minor metabolite of altizide and hydrochlorothiazide and could be detected up to 120 h. The hydrolysis product, 4-amino-6-chloro-1,3 benzenedisulphonamide, was detectable 120 h after administration, with concentrations at least 10 times higher than the parent drug. Concentrations ranged between 41-239 and 60-287 ng/mL after altizide and hydrochlorothiazide administration, respectively. The study shows that 4-amino-6-chloro-1,3 benzenedisulphonamide is an important target compound for the long time detection of thiazide diuretics in urine. PMID- 19187940 TI - Use of proteomics for design of a tailored host cell for highly efficient protein purification. AB - After some initial optimization, a downstream process comprised of one or several chromatography steps removes the majority of the host proteins and achieves a reasonable degree of purification. The separation of remaining contaminant proteins from the target protein could become very difficult and costly due to their similar physicochemical properties. In this paper we describe a highly efficient strategy, based on proteomic analysis and elution chromatography, by which a protein of interest may be isolated from copurifying contaminants. Mutant strains of Escherichia coli were prepared that are deficient in three prevalent host proteins found in a strategic fraction of an elution profile of nickel immobilized affinity chromatography. Recombinant green fluorescent protein (GFPuv) served as a model protein and its elution was directed to this optimized fraction with an N-terminus hexahistidine tag (his(6)), thereby easing its recovery. We demonstrate that proteomic data can facilitate the rational engineering of host cell expressing the target protein and the design of an efficient process for its purification. PMID- 19187941 TI - Hydrophilic interaction liquid chromatography with alcohol as a weak eluent. AB - There has been a significant increase of interest in polar compound separation by hydrophilic interaction liquid chromatography (HILIC), in which acetonitrile is mostly used as a weak eluent. Although replacing acetonitrile with alcohols as organic modifiers has been previously reported, the separation mechanism was poorly understood. In this paper we explored the separation mechanism through the method development for the analysis of the trace amounts of polar and basic hydrazines, which were genotoxic in nature. Separation parameters such as the type and concentration of alcohol, acid modifier, and buffer in mobile phase as well as the choice of stationary phase and column temperature were studied. The data indicated that both electrostatic and hydrophilic interactions contributed to the retention and separation of the hydrazines. The results presented here provide insight into the adjustment of the retention and separation of analytes in HILIC mode with alcohol as a weak eluent. The optimized HILIC method coupled with chemiluminescent nitrogen detection (CLND) is simple and sensitive (reporting limit at 0.02%) and was applied to simultaneous analysis of hydrazine and 1,1-dimethylhydrazine in a pharmaceutical intermediate. PMID- 19187942 TI - Histone deacetylase inhibitors and candidate gene expression: An in vivo and in vitro approach to studying chromatin remodeling in a clinical population. AB - OBJECTIVE: The emerging field of psychiatric epigenetics is constrained by the dearth of research methods feasible in living patients. With this focus, we report on two separate approaches, one in vitro and one in vivo, developed in our laboratory. METHOD: In the first approach, we isolated lymphocytes from 12 subjects and cultured their cells with either 0.7 mM valproic acid (VPA), 100 nM Trichostatin A (TSA), or DMSO (control) for 24h based upon previous dose response experiments. We then measured GAD67 mRNA expression using realtime RT-PCR, total acetylated histone 3 (H3K9,K14ac) levels using Western blot analysis, and attachment of H3K9,K14ac to the GAD67 promoter using ChIP. In the second approach, we measured GAD67 mRNA and total H3K9,K14ac levels in lymphocytes from 11 schizophrenia and 7 bipolar patients before and after 4 weeks of clinical treatment with Depakote ER (VPA). RESULTS: In the first approach, VPA induced a 383% increase in GAD67 mRNA, an 89% increase in total H3K9,K14ac levels, and a 482% increase in H3K9,K14ac attachment to the GAD67 promoter. TSA induced comparable changes on all measures. In the second approach, bipolar subjects had significantly higher baseline levels of H3K9,K14ac compared to subjects with schizophrenia. Subjects with clinically relevant serum levels of VPA (> or = 65 microg/mL) showed a significant increase in GAD67 mRNA expression. CONCLUSIONS: Our results utilizing two separate approaches for examining chromatin remodeling in real clinical time provide possible means to investigate epigenetic events in living patients. PMID- 19187943 TI - Bilateral ophthalmoplegia due to symmetric cavernous sinus metastasis from gastric adenocarcinoma. AB - We report a patient with rapidly progressive bilateral total ophthalmoplegia due to bilateral cavernous sinus metastasis from gastric adenocarcinoma. Among a variety of differential diagnoses, etiology of oculomotoric disorders and ophthalmoplegia includes the affection of the cranial nerves III, IV, and VI in the cavernous sinus. Unilateral metastasis in the cavernous sinus occurs quite frequently, however, a bilateral tumor infiltration of both cavernous sinus is extremely rare. Our patient impressively demonstrates the relevance of this differential diagnosis of bilateral ophthalmoplegia. Repeated CCTs and cMRIs were required to find the diagnosis and finally start a therapy, demonstrating that even with advanced neuroradiological techniques, repetition of imaging within short intervals can be necessary to detect rapidly developing metastatic infiltrations as early as possible. PMID- 19187944 TI - Impaired regulation of electron transport chain subunit genes by nuclear respiratory factor 2 in multiple sclerosis. AB - Multiple sclerosis (MS) is an inflammatory neurodegenerative disease. Recently, decreased expression of nuclear encoded electron transport chain genes was found in neurons in MS cortex. To understand the transcriptional mechanisms responsible for the coordinate down regulation of these genes, we performed electrophoretic mobility shifts with nuclear extracts isolated from gray matter from nonlesion areas of postmortem MS and control cortex. Nine tissue blocks from eight different MS brains and six matched control blocks from five control brains were analyzed. We identified a decrease in a transcription factor complex containing nuclear respiratory factor 2 (NRF-2) in nuclear extracts isolated from MS cortex. This decrease is correlated with decreased expression of electron transport chain subunit genes and increased oxidative damage measured by increased anti nitrotyrosine immunoreactivity. We conclude that in MS cortex a chronic increase in oxidative stress leads to aberrant regulation of transcription of genes involved in energy metabolism. PMID- 19187945 TI - Meiofauna with special reference to nematodes in trawling ground of subtropical Hong Kong. PMID- 19187946 TI - Dengue and concurrent urinary infection in a tertiary care hospital in Delhi. PMID- 19187947 TI - What about health system strengthening and the internal brain drain? PMID- 19187949 TI - Composting rice straw with sewage sludge and compost effects on the soil-plant system. AB - Composting organic residue is an interesting alternative to recycling waste as the compost obtained may be used as organic fertilizer. This study aims to assess the composting process of rice straw and sewage sludge on a pilot-scale, to evaluate both the quality of the composts obtained and the effects of applying such compost on soil properties and plant development in pot experiments. Two piles, with shredded and non-shredded rice straw, were composted as static piles with passive aeration. Throughout the composting process, a number of parameters were determined, e.g. colour, temperature, moisture, pH, electrical conductivity, organic matter, C/N ratio, humification index, cation exchange capacity, chemical oxygen demand, and germination index. Moreover, sandy and clayey soils were amended with different doses of mature compost and strewed with barley in pot experiments. The results show that compost made from shredded rice straw reached the temperatures required to maximise product sanitisation, and that the parameters indicating compost maturity were all positive; however, the humification index and NH(4) content were more selective. Therefore, using compost-amended soils at a dose of 34 Mg ha(-1) for sandy soil, and of 11 Mg ha( 1) for clayey soil improves soil properties and the growth of Hordeum vulgare plants. Under there conditions, the only limiting factor of agronomic compost utilisation was the increased soil salinity. PMID- 19187948 TI - Human papillomavirus type 16 E2 and E6 are RNA-binding proteins and inhibit in vitro splicing of pre-mRNAs with suboptimal splice sites. AB - Human papillomavirus type 16 (HPV16) genome expresses six regulatory proteins (E1, E2, E4, E5, E6, and E7) which regulate viral DNA replication, gene expression, and cell function. We expressed HPV16 E2, E4, E6, and E7 from bacteria as GST fusion proteins and examined their possible functions in RNA splicing. Both HPV16 E2, a viral transactivator protein, and E6, a viral oncoprotein, inhibited splicing of pre-mRNAs containing an intron with suboptimal splice sites, whereas HPV5 E2 did not. The N-terminal half and the hinge region of HPV16 E2 as well as the N-terminal and central portions of HPV16 E6 are responsible for the suppression. HPV16 E2 interacts with pre-mRNAs through its C terminal DNA-binding domain. HPV16 E6 binds pre-mRNAs via nuclear localization signal (NLS3) in its C-terminal half. Low-risk HPV6 E6, a cytoplasmic protein, does not bind RNA. Notably, both HPV16 E2 and E6 selectively bind to the intron region of pre-mRNAs and interact with a subset of cellular SR proteins. Together, these findings suggest that HPV16 E2 and E6 are RNA binding proteins and might play roles in posttranscriptional regulation during virus infection. PMID- 19187950 TI - Mortality due to lung, laryngeal and bladder cancer in towns lying in the vicinity of combustion installations. AB - BACKGROUND: Installations that burn fossil fuels to generate power may represent a health problem due to the toxic substances which they release into the environment. OBJECTIVES: To investigate whether there might be excess mortality due to tumors of lung, larynx and bladder in the population residing near Spanish combustion installations included in the European Pollutant Emission Register. METHODS: Ecologic study designed to model sex-specific standardized mortality ratios for the above three tumors in Spanish towns, over the period 1994-2003. Population exposure to pollution was estimated on the basis of distance from town of residence to pollution source. Using mixed Poisson regression models, we analyzed: risk of dying from cancer in a 5-kilometer zone around installations that commenced operations before 1990; effect of type of fuel used; and risk gradient within a 50-kilometer radius of such installations. RESULTS: Excess mortality (relative risk, 95% confidence interval) was detected in the vicinity of pre-1990 installations for lung cancer (1.066, 1.041-1.091 in the overall population; 1.084, 1.057-1.111 in men), and laryngeal cancer among men (1.067, 0.992-1.148). Lung cancer displayed excess mortality for all types of fuel used, whereas in laryngeal and bladder cancer, the excess was associated with coal fired industries. There was a risk gradient effect in the proximity of a number of installations. CONCLUSIONS: Our results could support the hypothesis of an association between risk of lung, laryngeal and bladder cancer mortality and proximity to Spanish combustion installations. PMID- 19187951 TI - Herbicide accumulation and evolution in reservoir sediments. AB - The aim of the present study was to understand the effect of reservoir configurations on sediment pesticide fate. Two dams were selected on the River Garonne, in southwest France: Carbonne and Golfech, both with reservoirs subject to accumulation of herbicide-contaminated sediment. They are situated upstream and downstream respectively of an agricultural and urban area: the Mid-Garonne. The results presented include pesticide concentrations and C/N ratios in the smaller sediment particles (<2 mm) and values of oxygenation and herbicide concentrations in the water. The dynamic behaviour of sediment in the reservoirs is discussed. The present study shows that the theoretical lifespan (weak remanence in vitro) and the results actually observed in the sediment are conflicting. Pesticide contamination in Carbonne indicates conservation, even accumulation, of herbicide molecules while in Golfech transformation processes clearly dominate. The hydromorphological position of Golfech reservoir, i.e. located at the junction of two rivers with contrasting hydrological regimes and very different oxygenation conditions, leads to accelerated pesticide desorption or degradation. Unfortunately, this configuration is rare. PMID- 19187952 TI - Urinary arsenic methylation capability and carotid atherosclerosis risk in subjects living in arsenicosis-hyperendemic areas in southwestern Taiwan. AB - Long-term exposure to inorganic arsenic from artesian drinking well water is associated with carotid atherosclerosis in the Blackfoot Disease (BFD) hyperendemic area in Taiwan. The current study examined the arsenic methylation capacity and its risk on carotid atherosclerosis. A total of 304 adults (158 men and 146 women) residing in the BFD-hyperendemic area were included. The extent of carotid atherosclerosis was assessed by duplex ultrasonography. Chronic arsenic exposure was estimated by an index of cumulative arsenic exposure (CAE) and the duration of artesian well water consumption. Urinary levels of inorganic arsenite [As(III)], arsenate [As(V)], monomethylarsonic acid [MMA(V)] and dimethylarsinic acid [DMA(V)] were determined by high performance liquid chromatography linked on line to a hydride generator and atomic absorption spectrometry (HPLC-HG-AAS). The percentage of arsenic species, primary methylation index [PMI=MMA(V)/(As(III)+As(V)] and secondary methylation index [SMI=DMA(V)/MMA(V)] were calculated and employed as indicators of arsenic methylation capacity. Results showed that women and younger subjects had a more efficient arsenic methylation capacity than did men and the elderly. Carotid atherosclerosis cases had a significantly greater percentage of MMA(V) [%MMA(V)] and a lower percentage of DMA [%DMA (V)] compared to controls. Subjects in the highest two tertiles of PMI with a median of CAE >0 mg/L-year had an odds ratio (OR) and a 95% confidence interval (CI) of carotid atherosclerosis of 2.61 and 0.98-6.90 compared to those in the highest two tertiles of PMI with a CAE=0 mg/L-year. We conclude that individuals with greater exposure to arsenic and lower capacity to methylate inorganic arsenic may be at a higher risk to carotid atherosclerosis. PMID- 19187953 TI - An evaluation of the reactivity of synthetic and natural apatites in the presence of aqueous metals. AB - Metal removal from contaminated effluents was examined following reaction with natural apatites of biological and geological origin or a synthetic hydroxylapatite (HAP). Mammalian meat and bone meal (MBM), a by-product from meat industry, was the biological apatite source. The effect of incineration on metal removal capacity of MBM and HAP was also examined. The reactivity of apatites for all tested metals (Pb, Cd, Cu and Zn) followed the general order: synthetic > biological > mineral. For all apatites tested, Pb was removed best and preferentially from multi-metal solutions. MBM and HAP (0.5 g solid) removed Pb completely from both highly concentrated single metal solutions (50 ml, 1000 mg/L Pb) and from multi-metal solutions (50 ml) with 100 mg/L each of Cd, Cu and Zn in addition to Pb. The incineration of MBM (725 degrees C and 850 degrees C) reduced significantly its capacity for removal of Zn (by 47%, from 56 mg/g to 9 mg/g) and Cd (by 38%, from 53 mg/g to 13 mg/g) in particular and to a lesser extent for Cu (by 14%, from 61 mg/g to 46 mg/g) while the removal of Pb was not affected (100 mg/g). The same pattern was observed for incinerated HAP. SEM and XRD analysis indicated that HAP reacted with the metals by precipitation of pure metal phosphates--Pb hydroxylapatite, Zn phosphate (hopeite), a Cd phosphate (identified only by ED-SEM) and Cu phosphate (libenthenite). PMID- 19187954 TI - Thromboelastography in patients with cerebral venous thrombosis. AB - INTRODUCTION: Cerebral venous thrombosis (CVT) is a rare presentation of venous thrombosis and has been associated with many conditions. In about 20% no risk factor is identified. The aim of this study was to assess the clot formation by thromboelastography (TEG) in patients with a history of CVT compared with healthy controls. MATERIALS AND METHODS: TEG analysis was performed on recalcified blood samples of 19 CVT patients from a single centre cohort and 1:1 sex/ age (+/-3 year) matched controls. Four TEG parameters were monitored: reaction time (r) to clot initiation, time to reach a 20 mm level of clot formation (K), slope angle alpha from r to K (alpha) and maximum vertical amplitude (MA). Patients were tested for thrombophilic defects, including deficiencies of antithrombin, protein C and protein S, factor V Leiden, prothrombin G20210A mutation, lupus anticoagulant, antiphospholipid antibodies, and high factor VIII levels. RESULTS: Thrombophilia testing identified a prothrombotic abnormality in 11 patients (58%). Sixteen patients (84%) had one or more transient risk factor. There were no significant differences in TEG parameters between CVT patients and controls, neither between the subgroup of patients with a thrombophilic defect and controls. Seven of all patients (37%), including 5 patients with abnormal thrombophilia testing, and 5 controls (26%) had one or more TEG hypercoagulable parameters. CONCLUSIONS: A persistent hypercoagulable state which could have predisposed to venous thrombosis in CVT patients and in the subgroup of patients with a thrombophilic defect could not be demonstrated by TEG. PMID- 19187956 TI - R-curve behavior and toughening mechanisms of resin-based dental composites: effects of hydration and post-cure heat treatment. AB - OBJECTIVES: To test the hypothesis that the fracture resistance of two different particulate resin composites degrade after water hydration and improve after post cure heat treatment, and to correlate those changes with salient failure micromechanisms. METHODS: Two composites with different filler morphology were selected, denoted microhybrid (Filtek Z250) and nanofill (Filtek Supreme plus). Following initial light curing, hydrated samples were aged in water for 60 days at room temperature while post-cured samples were heat treated at 120 degrees C for 90 min. Fracture resistance was assessed using fracture resistance curves (R curves) utilizing pre-cracked compact tension, C(T), specimens. The flexural strength of the hydrated composites also was evaluated in four-point bending using unnotched beams. Scanning electron microscopy (SEM) of crack paths and fracture surfaces was performed to determine the micromechanisms of fracture and toughening. The results were compared by two-way ANOVA and Tukey's multiple comparison test (p< or =0.05). RESULTS: SEM observations revealed a predominantly interparticle matrix crack path for all cases except the hydrated nanofill composite, which showed evidence of particle matrix debonding. Hydration lowered the strength for both composites and the peak toughness for the nanofill composite. The strength decrease was attributed to resin matrix plasticization and hydrolytic degradation in both cases, with additional interfacial degradation causing a larger strength decline and concomitant peak toughness decrease in the nanofill composite. The post-cure heat treatment noticeably changed the R-curve shape causing the peak toughness to be reached after shorter amounts of crack extension. Such changes help explain the increases in strength reported in other studies and is attributed to improved resin matrix properties. SIGNIFICANCE: Results from this study provide new insight into the micromechanisms of fracture in resin-based dental composites which should aid the future development and improvement of these materials. PMID- 19187955 TI - Graded structures for damage resistant and aesthetic all-ceramic restorations. AB - OBJECTIVES: Clinical studies revealed several performance deficiencies with alumina- and zirconia-based all-ceramic restorations: fracture; poor aesthetic properties of ceramic cores (particularly zirconia cores); and difficulty in achieving a strong ceramic-resin-based cement bond. We aim to address these issues by developing a functionally graded glass/zirconia/glass (G/Z/G) structure with improved damage resistance, aesthetics, and cementation properties. METHODS: Using a glass powder composition developed in our laboratory and a commercial fine zirconia powder, we have successfully fabricated functionally graded G/Z/G structures. The microstructures of G/Z/G were examined utilizing a scanning electron microscopy (SEM). The crystalline phases present in G/Z/G were identified by X-ray diffraction (XRD). Young's modulus and hardness of G/Z/G were derived from nanoindentations. Critical loads for cementation radial fracture in G/Z/G plates (20mmx20mm, 1.5 or 0.4mm thick) bonded to polycarbonate substrates were determined by loading with a spherical indenter. Parallel studies were conducted on homogeneous yttria-stabilized tetragonal zirconia polycrystal (Y TZP) controls. RESULTS: The G/Z/G structure consists of an outer surface aesthetic glass layer, a graded glass-Y-TZP layer, and a dense Y-TZP interior. The Young's modulus and hardness increase from surface to interior following power-law relations. For G/Z/G plates of 1.5 and 0.4mm thick, critical loads for cementation radial fracture were 1990+/-107N (mean+/-S.D., n=6) and 227+/-20N (mean+/-S.D., n=6), respectively, which were approximately 30 and 50% higher than those for their monolithic Y-TZP counterparts (1388+/-90N for 1.5mm and 113+/-10N for 0.4mm thick; mean+/-S.D., n=6 for each thickness). A 1-sample t-test revealed significant difference (p<0.001) in critical loads for radial fracture of G/Z/G and homogeneous Y-TZP for both specimen thicknesses. SIGNIFICANCE: Our results indicate that functionally graded G/Z/G structures exhibit improved damage resistance, aesthetics, and potentially cementation properties compared to homogeneous Y-TZP. PMID- 19187957 TI - The barnacle Balanus amphitrite as a biomonitor for Cd: radiolabelled experiments. AB - Radiolabelled experiments were carried out to measure necessary parameters in the development of a biodynamic ecotoxicological simulation model of Cd accumulation in the barnacle biomonitor Balanus amphitrite. The Cd uptake rate constant from the dissolved phase, the Cd assimilation efficiency (AE) from suspended particulate matter (SPM) and the efflux rate constant were obtained using (109)Cd. A Cd uptake rate constant from the dissolved phase (k(u)) of 0.0072 Lg( 1)h(-1) was determined for the barnacle under environmentally realistic dissolved Cd concentrations (maximum of 400 ng L(-1)). Cd AE from SPM was determined from the barnacle feeding on SPM with low and high chl a concentrations, resulting in AEs of 39.0% and 48.7%, respectively, and an efflux rate of 0.0072 d(-1). The difference between the AEs resulted from differences in chl a:SPM ratios suggesting a general tendency of higher AE when SPM is enriched with chl a. These results reinforce that the accuracy of ecotoxicological models for metal accumulation in organisms depends on how representative the selected food items are of the organism's natural diet. PMID- 19187958 TI - Mesenchymal stem cell proliferation and differentiation on an injectable calcium phosphate-chitosan composite scaffold. AB - Calcium phosphate cement (CPC) can be molded or injected to form a scaffold in situ, has excellent osteoconductivity, and can be resorbed and replaced by new bone. However, its low strength limits CPC to non-stress-bearing repairs. Chitosan could be used to reinforce CPC, but mesenchymal stem cell (MSC) interactions with CPC-chitosan scaffold have not been examined. The objective of this study was to investigate MSC proliferation and osteogenic differentiation on high-strength CPC-chitosan scaffold. MSCs were harvested from rat bone marrow. At CPC powder/liquid (P/L) mass ratio of 2, flexural strength (mean+/-sd; n=5) was (10.0+/-1.1) MPa for CPC-chitosan, higher than (3.7+/-0.6) MPa for CPC (p<0.05). At P/L of 3, strength was (15.7+/-1.7)MPa for CPC-chitosan, higher than (10.2+/ 1.8)MPa for CPC (p<0.05). Percentage of live MSCs attaching to scaffolds increased from 85% at 1 day to 99% at 14 days. There were (180+/-37) cells/mm(2) on scaffold at 1 day; cells proliferated to (1808+/-317) cells/mm(2) at 14 days. SEM showed MSCs with healthy spreading and anchored on nano-apatite crystals via cytoplasmic processes. Alkaline phosphatase activity (ALP) was (557+/-171) (pNPP mM/min)/(microg DNA) for MSCs on CPC-chitosan, higher than (159+/-47) on CPC (p<0.05). Both were higher than (35+/-32) of baseline ALP for undifferentiated MSCs on tissue-culture plastic (p<0.05). In summary, CPC-chitosan scaffold had higher strength than CPC. MSC proliferation on CPC-chitosan matched that of the FDA-approved CPC control. MSCs on the scaffolds differentiated down the osteogenic lineage and expressed high levels of bone marker ALP. Hence, the stronger CPC-chitosan scaffold may be useful for stem cell-based bone regeneration in moderate load-bearing maxillofacial and orthopedic applications. PMID- 19187959 TI - F281, synthetic agonist of the sigma-2 receptor, induces Ca2+ efflux from the endoplasmic reticulum and mitochondria in SK-N-SH cells. AB - We demonstrate that F281, a synthetic agonist of the sigma-2 receptor (s2R), induces a non transient increase in intracellular [Ca(2+)] ([Ca(2+)](i)) and cell death in SK-N-SH cells. Sigma receptors are classified into two subtypes, with different molecular weight and tissue distribution. While the sigma-1 receptor has been cloned, the s2r is less characterized and its physiological ligand and role need further investigation. In tumour cell lines, synthetic agonists of the s2R trigger apoptosis and modulate [Ca(2+)](i). In particular, CB-64D induces a Ca(2+) response while PB28 supresses Ca(2+) signalling. We have recently synthesized F281, by replacing the 5-methoxytetraline moiety of PB28 with a carbazole nucleus. Although this bioisosteric substitution should not affect the ligand affinity at the receptor, F281 (after 24h incubation) was more cytotoxic than PB28 (EC(50) values 65.4nM and 8.13 microM, respectively) in SK-N-SH cells. We used the fluorescent probes fura-2, rhod-2 and JC-1. F281 mobilizes Ca(2+) from mitochondria and from the endoplasmic reticulum, by opening its inositol 1,4,5-trisphosphate receptor; Ca(2+)-entry through the channels activated by store depletion was also observed. After the increase in [Ca(2+)](i) and within 10 min, we observed a sudden drop in metabolic activity and intracellular [ATP] leading to cell death. PMID- 19187960 TI - Twice daily fludarabine/Ara-C associated to idarubicin, G-CSF and ATRA is an effective salvage regimen in non-promyelocytic acute myeloid leukemia. AB - Preclinical data suggest that all-trans retinoic acid (ATRA) synergizing with granulocyte colony stimulating factor (G-CSF), can improve the effectiveness of chemotherapy in acute myeloid leukemia (AML). Fludarabine 15 mg/m(2) is the minimum dose able to optimize intensification with fludarabine-arabinosylcytosine regimen. In this study 52 patients with relapsed/refractory AML obtained a complete remission (CR) rate of 69.2% after FLAIRG regimen (Fludarabine and arabinosylcytosine twice daily, idarubicin, G-CSF, ATRA). This schedule resulted effective and tolerable enabling 53% of the responding patients to receive transplant procedure. FLAIRG regimen could be proposed as a "bridge" to transplant treatment in this poor risk setting. PMID- 19187961 TI - Biochemical responses of the shore crab (Carcinus maenas) in a eutrophic and metal-contaminated coastal system (Obidos lagoon, Portugal). AB - A eutrophic and metal-contaminated coastal system (Obidos lagoon, Portugal) was monitored combining water/sediment quality parameters and Carcinus maenas biomarkers (accumulated metals, oxidative stress and biotransformation responses). Two confined branches (Barrosa and Bom-Sucesso) were surveyed and compared with a reference area. Both crab genders from Barrosa exhibited activation of hepatopancreas CAT, GPx and GST, pointing out this area as the major impacted in the lagoon. Females captured at Barrosa were more vulnerable to peroxidative damage while only males showed decreased EROD activity, reinforcing gender specificities. In general, responses were not directly attributed to metals in hepatopancreas, as supported by Principal Component Analysis (PCA). However, higher metals (Ni, Cu, Cd) and nutrients levels registered in Barrosa water were associated with the observed oxidative stress responses by PCA. Despite the difficulty to establish cause-effect relationships due to the co occurrence of various stressors and their interactions, the adopted integrated monitoring strategy appears to be promising. PMID- 19187962 TI - Combined effects of Psoralens and ultraviolet-B on growth, pigmentation and biochemical parameters of Abelmoschus esculentus L. AB - The effects of pre-treatment of Psoralens (furocoumarin compounds) and supplemental ultraviolet-B (sUV-B) were studied on plant growth, photosynthetic and non-photosynthetic pigments, protein, phenylalanine ammonia lyase (PAL) activity and antioxidative defense potential as well as their ultimate effects on biomass production in Abelmoschus esculentus L. (Okra) plants. Psoralens are capable of absorbing radiant energy and stimulating the pigmentation of human skin when photo-activated in presence of UV-A or UV-B making them beneficial in the treatment of vitilago. Pre-treatment of Psoralens against sUV-B (pUV-B), stimulates higher production of UV-B protective pigments (flavonoids and carotenoids) and helps in maintaining its biomass against UV-B stress. Antioxidative defense system in the test plant was activated by combined treatment of Psoralens and sUV-B as evidenced by the enhanced activity of enzymatic (ascorbate peroxidase-APX, superoxide dismutase-SOD, POX) and non enzymatic (ascorbic acid and phenol) antioxidants. Individual treatments of Psoralens and sUV-B showed inhibitory effect on various morphological traits i.e. reduction in plant height, leaf area and ultimately on biomass production. Our results clearly indicated that adverse effect of sUV-B on biomass production was ameliorated by pre- treatment with Psoralens. PMID- 19187963 TI - Comparative analysis of four lipoproteins from Mycoplasma mycoides subsp. mycoides Small Colony identifies LppA as a major T-cell antigen. AB - Control of contagious bovine pleuropneumonia (CBPP), caused by Mycoplasma mycoides subsp. mycoides Small Colony (MmmSC), remains an important goal in Africa. Subunit vaccines triggering B and T-cell responses could represent a promising approach. To this aim, the T-cell immunogenicity of four MmmSC lipoproteins (LppA, LppB, LppC and LppQ), present in African strains and able to elicit humoral response, was evaluated. In vitro assays revealed that only LppA was recognized by lymph node lymphocytes taken from three cattle, 3 weeks after MmmSC exposure. Maintenance of the LppA-specific response, relying on CD4 T-cells and IFN gamma production, was then demonstrated 1 year after infection. LppA is thus an important target for the CD4 T-cells generated early after MmmSC infection and persisting in the lymph nodes of recovered cattle. Its role as a protective antigen and ability to in vivo trigger both arms of the host immune response remain to be evaluated. PMID- 19187964 TI - Monoclonal antibodies specific to Cucumber mosaic virus coat protein possess DNA hydrolyzing activity. AB - Monoclonal antibodies (mAbs) specific to Cucumber mosaic virus coat protein (CMV CP) were designed from cDNA and deduced amino acid sequences of the light chain genes of 10 out of 14 different hybridoma cell lines. Ten of these mAbs revealed a very restricted germline family VkappaII, within which gene bd2 has identical amino acid sequences with VIPase and an i41SL 1-2 catalytic antibody light chain, both of which possess peptidase activity. Four out of the 14 mAbs illustrated another germline family VkappaIA, within which gene bb1.1 had high homology with BV04-01 light chain mAb, which hydrolyses ssDNA. Interestingly, our mAbs showed DNA-hydrolytic activity at an optimum pH of 4-5, which is a typical pattern of autoimmune diseases in which autoantibodies hydrolyze supercoiled plasmid DNA. This is the first evidence ever that CMV-CP could stimulate catalytic antibodies, which have an identical sequence homology with autoantibodies. Furthermore, the CMV-CP-specific mAbs will be important for isolating antibodies specific to the CPs of bacteria, viruses, cancer cells, etc. that could be used for medical therapy. PMID- 19187965 TI - Increased B cell deletion and significantly reduced auto-antibody titre due to premature expression of human complement receptor 2 (CR2, CD21). AB - The involvement of complement receptor 2 (CR2) in B cell tolerance and autoimmune disease has been revealed over the past decade or so. Our previous studies have established that mice prematurely expressing human CR2 under the control of a lambda light chain promoter (in particular the hCR2(high) line) have a marked deficit in their immune response to various antigens and fail to develop collagen induced arthritis. This phenotype appears to be the result of irreversible changes in B cell signalling pathways and suggested that hCR2 expressing mice are protected from developing autoimmune disease. To test this hypothesis, we examined the ability of the hCR2 to block the development of spontaneous autoimmune disease on the C57BL/6j-Fas(lpr/)Fas(lpr) (B6(lpr)) background. We found that expression of hCR2 on the B6(lpr) background resulted in a significant reduction in levels of anti-nuclear antibodies (ANA) generated as mice aged but the levels of ANA were still higher than those found in age matched C57BL/6j (B6) mice. B cells from hCR2(high) mice were found to display a higher baseline level of apoptosis, whether analysed ex vivo or after in vitro culture, than their B6 counterparts and this was apparently linked to both surface IgM expression by the B cells and C3 levels in the mice. Our data also provides evidence that B cell survival in the presence of hCR2 is heavily modified by the background strain of the mouse. Overall, we have demonstrated that mice expressing hCR2 on their B cells during bone marrow development display a higher degree of apoptosis which may lead to a deletion of autoreactive B cells and be protective against the development of autoimmune disease. PMID- 19187966 TI - Chicken heterophils are recruited to the site of Salmonella infection and release antibacterial mature Cathelicidin-2 upon stimulation with LPS. AB - The biological functions of avian cathelicidins are poorly defined. In mammals, cathelicidins have shown to possess potent broad-range antimicrobial activity as well as immunomodulatory activities. Therefore, we investigated the microbicidal activities and localization of Cathelicidin-2 in non-infected and Salmonella challenged broiler chickens. Using immunohistochemistry, Cathelicidin-2 was shown to be abundantly present in heterophils, localized in the large rod-shaped granules, but absent in other peripheral blood cells and intestinal epithelial cells. Cathelicidin-2 synthesis was observed to be initiated at the early promyelocyte stage. Considerable infiltration of Cathelicidin-2 containing heterophils was observed in the jejunum of Salmonella enteritidis-challenged broilers within 8 h post-infection. Heterophils were shown to release mature Cathelicidin-2 peptide upon stimulation with Salmonella-derived LPS in a time dependent way. Processing of the Cathelicidin-2 precursor was mediated by serine proteases with a divalent cation dependency. Cathelicidin-2 peptide showed potent bactericidal and fungicidal activity against all tested microorganisms, including chicken-specific Salmonella isolates. These results underscore the importance of avian heterophils as a first line of defence against invading pathogens and implicate that via heterophil-mediated release, cathelicidins may greatly contribute to avian innate immunity. PMID- 19187967 TI - Structural, magnetic, electrochemical, catalytic, DNA binding and cleavage studies of new macrocyclic binuclear copper(II) complexes. AB - The macrocyclic symmetrical and a series of unsymmetrical binuclear copper(II) complexes have been synthesized by using mononuclear complex [CuL] [3,3'-((1E,7E) 3,6-dioxa-2,7-diazaocta-1,7-diene-1,8-diyl)bis(3-formyl-5-methyl-2 diolato)copper(II)]. Another compartment of the [CuL] have been condensed with various diamines like 1,2-bis(aminooxy)ethane (L(1)), 1,2-diamino ethane(L(2a)), 1,3-diamino propane(L(2b)), 1,4-diamino butane(L(2c)), 1,2-diamino benzene(L(2d)), 1,8-diamino naphthalene(L(2e)) and characterized by elemental, spectroscopic, and X-ray crystallographic methods. The influence of the coordination geometry and the ring size of the binucleating ligands on the electronic, redox, magnetic, catecholase activity, DNA binding and cleavage properties have been studied. The molecular structures of the symmetrical binuclear complex [Cu(2)L(1)(H(2)O)(2)](ClO(4))(2) (1) and unsymmetrical binuclear complex [Cu(2)L(2b)(ClO(4))(H(2)O)]ClO(4) (2b) were determined by X-ray crystallography. Both of them were discrete binuclear species in which each Cu(II) ions are in distorted square pyramid. The Cu...Cu distances vary from 3.0308 (2b) to 3.0361 A (1). Electrochemical studies evidenced that two quasi reversible one electron-transfer reduction waves (E(pc)(1)) -0.91 to -1.01 V, (E(pc)(2)) -1.26 to -1.55 V) for binuclear complexes are obtained in the cathodic region. Cryomagnetic investigation of the binuclear complexes reveals a weak antiferromagnetic spin exchange interaction between the Cu(II) ions within the complexes (-2J=104.4-127.5 cm(-1)). The initial rate (V(in)) for the oxidation of 3,5-di-tert-butylcatechol to o-quinone by the binuclear Cu(II)complexes are in the range 3.6 x 10(-5) to 7.3 x 10(-5)Ms(-1). The binuclear Cu(II) complexes are avid binders to calf thymus DNA. The complexes display significant oxidative cleavage of circular plasmid pBR322 DNA in the presence of mercaptoethanol using the singlet oxygen as a reactive species. The aromatic diamine condensed macrocyclic ligands of copper(II) complexes display better DNA interaction and significant chemical nuclease activity than the aliphatic diamine condensed macrocyclic Cu(II) complexes. PMID- 19187968 TI - DNA interaction studies of tridentate bridged Ru(II)-Pt(II) mixed-metal supramolecules. AB - Reported herein are studies of the concentration and temperature dependent interactions with DNA of the stereochemically defined mixed-metal supramolecular complexes, [(tpy)Ru(tppz)PtCl](PF(6))(3) and [ClPt(tppz)Ru(tppz)PtCl](PF(6))(4) (tpy=2,2':6',2''-terpyridine and tppz=2,3,5,6-tetrakis(2-pyridyl)pyrazine). These metal complexes couple a ruthenium based light absorber (LA) to the bioactive platinum sites (BAS) using a tridentate bridging ligand (BL). The complexes exhibit intense Ru-->tppz(pi*) metal to ligand charge transfer (MLCT) transitions in the visible region and adopt a square planar geometry around the Pt(II) center. The effect of incubating these metal complexes with DNA on the subsequent migration of DNA through an agarose gel was found to be more dramatic than that observed for the well known anticancer drug, cis-[Pt(NH(3))(2)Cl(2)] (cisplatin). This effect was enhanced with increased incubation temperature. Unwinding of supercoiled plasmid DNA was found to be more pronounced for the trimetallic complex, [ClPt(tppz)Ru(tppz)PtCl](PF(6))(4), than for the bimetallic complex, [(tpy)Ru(tppz)PtCl](PF(6))(3). PMID- 19187969 TI - Platinum-based complexes of bioactive 3-(5-nitrofuryl)acroleine thiosemicarbazones showing anti-Trypanosoma cruzi activity. AB - Eight new platinum(II) complexes with 3-(5-nitrofuryl)acroleine thiosemicarbazones showing anti-trypanosomal activity were synthesized, characterized and in vitro evaluated. Most of the complexes showed IC(50) values in the micromolar range against two different strains of Trypanosoma cruzi, causative agent of Chagas disease (American Trypanosomiasis). In addition, most of the newly developed complexes, together with the analogous platinum 5 nitrofuraldehyde containing thiosemicarbazones previously reported, resulted more active than the reference trypanocidal drug nifurtimox on the infective trypomastigote form of the parasite. Their capacity to produce free radicals that could lead to parasite death was evaluated by ESR experiments in the parasite and by respiration measurements. Compounds were tested for their DNA interaction ability. Results showed that some of the compounds could act as dual inhibitors in the parasite, through production of toxic free radicals and interaction with DNA. All the results were compared with those previously reported for the free ligands, the analogous palladium(II) compounds and the previously reported series of platinum(II) compounds. PMID- 19187970 TI - Recent and lifetime utilization of health care services by children and adolescent suicide victims: a case-control study. AB - OBJECTIVE: In the present paper we describe a case-control study on the utilization of health care services prior to suicide (across different time periods) among children and adolescents aged 11 to 18 years in the Province of Quebec, Canada and matched healthy controls. METHOD: Utilization of services (i.e., contact with general practitioners, mental health professionals, psychiatrists and/or youth protection groups) was examined at different time periods in 55 child and adolescent suicide victims and 54 matched community controls using proxy-based interviews and questionnaires. In addition, we examined the rates of detection of psychopathology by health care professionals, the use of psychotropic medications and the subjects' compliance with treatment. RESULTS: Although more than 90% of child and adolescent suicide completers in our sample suffered from mental disorders, a significant proportion of them were left without appropriate healthcare support (including psychiatric consultation) in the period preceding their suicide. Also, 20% of suicide completers and no control subject made prior suicide attempts. More specifically, over two-thirds of suicide completers had no treatment contact within the month prior to the completion, while only 12.7% (n=7) of them were in contact with psychiatric services during that same period. Moreover, 56.4% (n=31) of the suicide completers had not been diagnosed as having a mental disorder at the time of their death, and 54.5% of the subjects' that received treatment (12 out of 22) were considered poorly compliant or not compliant at all according to their medical/psychosocial records. Finally, we also found that females seemed to have more psychiatric and mental health service contacts in the past month, that subjects with depressive and anxious disorders received more psychiatric and general mental health services in the past year, and that past month hospitalization was more often associated with alcohol abuse and psychosis. LIMITATIONS: Relatively small sample size, and cross-sectional design. CONCLUSIONS: Our findings indicate the need for an overall increase in the rates of healthcare services delivered to young subjects at risk for suicide, as well as better training of health professionals in detecting and treating youth psychopathology. PMID- 19187971 TI - Autoantibody to dihydropyridine receptor in myasthenia gravis. AB - To investigate autoantibodies related to excitation-contraction (E-C) coupling in patients with myasthenia gravis (MG), we developed a novel method to detect autoantibodies against dihydropyridine receptor (DHPR). Using this method, we detected DHPR antibody in 37% (11 out of 30) of MG patients with thymoma. Antibodies were not detected in normal nor disease controls. The titer of DHPR antibodies showed no significant correlation with autoantibodies to acetylcholine nor ryanodine receptors. The DHPR antibody is another marker for thymoma in MG, and it might have some role in clinical symptoms related to E-C coupling. PMID- 19187972 TI - Intravenous tolerance modulates macrophage classical activation and antigen presentation in experimental autoimmune encephalomyelitis. AB - Macrophages act as the first line of self defense by mounting an inflammatory response to antigen and as antigen presenting cells to initiate the adaptive immune response. Inhibition of macrophage activation is one of the possible approaches to modulate inflammation. Intravenous (i.v.) tolerance has proved to be an effective method for ameliorating experimental autoimmune diseases. Whether macrophages are involved in tolerance induction is still largely undefined. In the present study we found that i.v. tolerance induction resulted in lower B7.1, B7.2 and MHC class II molecules, and reduced phagocytosis by both peritoneal macrophages and adherent splenocytes. Macrophages from tolerized mice were associated with a significantly impaired response of MOG-sensitized T cells to MOG. Macrophages from tolerized mice produced low levels of pro-inflammatory molecules IL-12, TNF-alpha, IL-1beta, RANTES and MCP-1 and high levels of IL-10 and TGF-beta. Administration of anti-TGF-beta led to a reduction of IL-10 in tolerized mice. Thus, i.v. tolerance inhibits macrophage classical activation and APC function, increases macrophage alternative activation and IL-10 and TGF-beta production. These cytokines, in turn, induce enhanced production of IL-10 in macrophages in MOG i.v. mice. PMID- 19187973 TI - Mutations in TMC1 contribute significantly to nonsyndromic autosomal recessive sensorineural hearing loss: a report of five novel mutations. AB - Genome wide homozygosity mapping using Affymetrix 10K arrays revealed the DFNB7/11 locus including the TMC1 gene in 5 of 35 Turkish families with autosomal recessive nonsyndromic severe to profound congenital or prelingual-onset sensorineural hearing loss (SNHL). Additional 51 families were later screened for co-segregation of the locus with the phenotype using microsatellite markers. GJB2 and mtDNA A1555G mutations were negative in probands from each family. Mutation analysis was performed in families showing co-segregation of autosomal recessive SNHL with haplotypes at the DFNB7/11 locus. A total of six different mutations in seven families were identified, including novel missense alterations, p.G444R (c.1330G>A), p.R445C (c.1333C>T), and p.I677T (c.2030T>C), one novel splice site mutation IVS6+2 T>A (c.64+2T>A), and a novel large deletion of approximately 31kb at the 3' region of the gene including exons 19-24, as well as a previously reported nonsense mutation, p.R34X (c.100C>T). All identified mutations co segregated with autosomal recessive SNHL in all families and were not found in Turkish hearing controls. These results expand the mutation spectrum of TMC1 with five novel mutations and provide data for the significant contribution of TMC1 mutations in hearing loss. PMID- 19187974 TI - 30 years of change in the world, science and medicine. PMID- 19187975 TI - Early hearing detection and intervention in South Africa. AB - Early hearing detection and intervention programs have become the standard of care to ensure optimal outcomes for infants with hearing loss, their families and society at large. The overwhelming majority of infants with congenital or early onset permanent bilateral hearing loss are however born in developing countries like South Africa where services are scarce and awareness poor. Despite its comparatively well-developed economic and reasonably developed health care infrastructure in sub-Saharan Africa, limited information on infant hearing loss and the status of early hearing detection and intervention has been available for South Africa. Recently however, an increasing number of initiatives and reports have highlighted the extent of infant hearing loss and the status of identification and intervention services offered in the country. This report provides a review of the available evidence on infant hearing loss and the status of current early hearing detection and intervention services in South Africa. PMID- 19187976 TI - Development of pediatric otolaryngology in North America. PMID- 19187977 TI - The importance of discerning shape in molecular pharmacology. AB - Shape is a fundamentally important molecular feature that often determines the fate of a compound in terms of molecular interactions with preferred and non preferred biological targets. Complementarity of binding in small-molecule protein, peptide-receptor, antigen-antibody and protein-protein interactions is the key to life and survival and also to targeting molecules with bioactivity. We review the application of shape in various biological systems such as substrate recognition, ligand specificity or selectivity and antibody recognition in the context of computational methods such as docking, quantitative structure-activity relationships, classification models and similarity-search algorithms. These in silico pharmacology methods have recently demonstrated the importance and applicability of determining molecular shape in drug discovery, virtual screening and predictive toxicology. The results from recently published studies show that shape and shape-based descriptors are at least as useful as other traditional molecular descriptors. PMID- 19187979 TI - Associative memory models: from the cell-assembly theory to biophysically detailed cortex simulations. AB - The second half of the past century saw the emergence of a theory of cortical associative memory function originating in Donald Hebb's hypotheses on activity dependent synaptic plasticity and cell-assembly formation and dynamics. This conceptual framework has today developed into a theory of attractor memory that brings together many experimental observations from different sources and levels of investigation into computational models displaying information-processing capabilities such as efficient associative memory and holistic perception. Here, we outline a development that might eventually lead to a neurobiologically grounded theory of cortical associative memory. PMID- 19187978 TI - STIM and Orai: the long-awaited constituents of store-operated calcium entry. AB - Rapid changes in cytosolic Ca(2+) concentrations [Ca(2+)](i) are the most commonly used signals in biology to regulate a whole host of cellular functions including contraction, secretion and gene activation. A widely utilized form of Ca(2+) influx is termed store-operated Ca(2+) entry (SOCE) owing to its control by the Ca(2+) content of the endoplasmic reticulum (ER). The underlying molecular mechanism of SOCE has eluded identification until recently when two groups of proteins, the ER Ca(2+) sensors stromal interaction molecule (STIM)1 and STIM2 and the plasma-membrane channels Orai1, Orai2 and Orai3, have been identified. These landmark discoveries have enabled impressive progress in clarifying how these proteins work in concert and what developmental and cellular processes require their participation most. As we begin to better understand the biology of the STIM and Orai proteins, the attention to the pharmacological tools to influence their functions quickly follow suit. Here, we briefly summarize recent developments in this exciting area of Ca(2+) signaling. PMID- 19187981 TI - Seven-year recurrence of left ventricular apical ballooning. AB - Left ventricular apical ballooning (Tako-tsubo syndrome) is an acute and rapidly reversible dysfunction of the left ventricle, triggered by a profound psychological stress. We describe an interesting case of very late recurrence. PMID- 19187980 TI - Nickel and binary metal mixture responses in Daphnia magna: molecular fingerprints and (sub)organismal effects. AB - The recent development of a custom cDNA microarray platform for one of the standard organisms in aquatic toxicology, Daphnia magna, opened up new ways to mechanistic insights of toxicological responses. In this study, the mRNA expression of several genes and (sub)organismal responses (Cellular Energy Allocation, growth) were assayed after short-term waterborne metal exposure. Microarray analysis of Ni-exposed daphnids revealed several affected functional gene classes, of which the largest ones were involved in different metabolic processes (mainly protein and chitin related processes), cuticula turnover, transport and signal transduction. Furthermore, transcription of genes involved in oxygen transport and heme metabolism (haemoglobin, delta-aminolevilunate synthase) was down-regulated. Applying a Partial Least Squares regression on nickel fingerprints and biochemical (sub)organismal parameters revealed a set of co-varying genes (haemoglobin, RNA terminal phosphate cyclase, a ribosomal protein and an "unknown" gene fragment). An inverse relationship was seen between the mRNA expression levels of different cuticula proteins and available energy reserves. In addition to the nickel exposure, daphnids were exposed to binary mixtures of nickel and cadmium or nickel and lead. Using multivariate analysis techniques, the mixture mRNA expression fingerprints (Ni2+ + Cd2+, Ni2+ + Pb2+) were compared to those of the single metal treatments (Ni2+, Cd2+, Pb2+). It was hypothesized that the molecular fingerprints of the mixtures would be additive combinations of the gene transcription profiles of the individual compounds present in the mixture. However, our results clearly showed additionally affected pathways after mixture treatment (e.g. additional affected genes involved in carbohydrate catabolic processes and proteolysis), indicating interactive molecular responses which are not merely the additive sum of the individual metals. These findings, although indicative of the complex nature of mixture toxicity evaluation, underline the potential of a toxicogenomics approach in gaining more mechanistic information on the effects of single compounds and mixtures. PMID- 19187982 TI - Optimizing long-term dual aspirin/clopidogrel therapy in acute coronary syndromes: when does the risk outweigh the benefit? AB - Most guidelines recommend indefinite use of aspirin in patients at increased atherothrombotic risk. Dual antiplatelet therapy (aspirin/clopidogrel) is significantly more effective than aspirin monotherapy in reducing cardiovascular risk in a number of patient populations. As a result, dual therapy is recommended in many patient groups, including those with acute coronary syndromes and those who have undergone percutaneous coronary intervention. The recommended duration of clopidogrel/aspirin treatment is generally less than one year and, in some cases, as little as one month. However, there is evidence from a range of patient populations that more prolonged clopidogrel/aspirin therapy may be more effective than short-term treatment in reducing cardiovascular risk in some patients. The effects of discontinuing clopidogrel after an event-free period of one year require examination in clinical trials. Trial data are also needed to guide management of patients in whom early antiplatelet withdrawal is being considered (e.g. those who require non-cardiac surgery). This review discusses the benefits and risks of long-term dual antiplatelet therapy in a range of clinical situations. PMID- 19187983 TI - Which application for systolic wall stress and circumferential fiber velocity shortening in echocardiography in the tissue Doppler imaging era? AB - We discuss in the paper the usefulness of old echocardiographic parameters including systolic wall stress and fiber velocity shortening in the tissue Doppler imaging era. PMID- 19187984 TI - Clinical and hemodynamic predictors of natriuretic peptide elevation among aortic valve disease patients. AB - We investigated the relative contribution of hemodynamic and clinical factors to serum natriuretic peptide elevation in seventy-one patients with either aortic stenosis or aortic regurgitation. We found that pulmonary hypertension, heart failure and renal failure are the most powerful independent predictors of natriuretic peptide elevation in patients with aortic valve disease, irrespective of the type or severity of valvular lesion itself. PMID- 19187985 TI - Steroid-induced crisis and rhabdomyolysis in a patient with pheochromocytoma: a case report and review. AB - A 66-year-old Japanese woman was urgently referred to our hospital. Two days prior to admission, her general practitioner began to administer prednisolone for treatment of a diagnosis of polymyalgia rheumatica. At the time of admission, laboratory results indicated multiorgan failure with rhabdomyolysis. Abdominal ultrasonography and computed tomography revealed a tumor in the right adrenal gland. On the same day, we measured serum and urine cathecholamines, which were markedly elevated. Additionally, magnetic resonance imaging revealed an adrenal mass and metaiodobenzylguanidine scintigraphy showed labeling of the tumor. Then, the patient underwent surgical resection of the tumor via laparoscopy. Histological examination confirmed the diagnosis of pheochromocytoma. One week after the operation, serum and urinary catecholamine levels returned to normal. The patient was discharged 10 days after the operation, and has remained stable at home. This report indicates that steroid should be avoided if possible in patients with pheochromocytoma. Furthermore, pheochromocytoma should be recalled as a differential diagnosis whenever patients take a sudden turn for the worse, or have acute uncontrollable hypertension following steroid administration and/or whenever patients present with unexplained rhabdomyolysis. PMID- 19187986 TI - Association of parental diabetes with overweight in Iranian children and adolescents. PMID- 19187987 TI - Rich and healthy-better than poor and sick? An empirical analysis of income, health, and the duration of the pension benefit spell. AB - I analyze the relationship between duration of the pension benefit spell and pension benefit claims from the German public pension system, with a special emphasis on differential results with respect to health. This relationship is crucial for a potential structural pattern of redistribution between different income and health groups, induced by the public pension system. Evidence for such redistribution from poor to rich is present for most of the specifications in my analysis. The most adequate specification is partially linear, does therefore not impose any parametrical restrictions between duration and benefit claims, and allows for potential endogeneity. The relationship I extract is clearly positive. I also find that redistribution from low-income individuals to individuals with high income is more pronounced the worse the health status is, despite longer benefit spells for pensioners in ill-health. PMID- 19187988 TI - Mammalian cell-based biosensors for pathogens and toxins. AB - Cell-based biosensors (CBBs) have emerged as powerful functional tools for the rapid detection of hazards and threats associated with food, agriculture, environment and biosecurity. CBBs detect the functional aspects of a host-hazard interaction and render an accurate estimation of the risks. Assessing hazard induced physiological responses, such as receptor-ligand interactions, signal transduction, gene expression, membrane damage, apoptosis and oncosis of living sensing organisms can provide insight into the basis of toxicity for a particular hazard. This review highlights the progress made in developing mammalian CBBs for pathogens and toxins, with special emphasis on multidisciplinary approaches that combine molecular biology and microbiology with methods used in physics and engineering, which led to the development of a three-dimensional cell-culture system and high-throughput screening employing optical and electrical systems. PMID- 19187989 TI - Classification of platelet concentrates: from pure platelet-rich plasma (P-PRP) to leucocyte- and platelet-rich fibrin (L-PRF). AB - The topical use of platelet concentrates is recent and its efficiency remains controversial. Several techniques for platelet concentrates are available; however, their applications have been confusing because each method leads to a different product with different biology and potential uses. Here, we present classification of the different platelet concentrates into four categories, depending on their leucocyte and fibrin content: pure platelet-rich plasma (P PRP), such as cell separator PRP, Vivostat PRF or Anitua's PRGF; leucocyte- and platelet-rich plasma (L-PRP), such as Curasan, Regen, Plateltex, SmartPReP, PCCS, Magellan or GPS PRP; pure plaletet-rich fibrin (P-PRF), such as Fibrinet; and leucocyte- and platelet-rich fibrin (L-PRF), such as Choukroun's PRF. This classification should help to elucidate successes and failures that have occurred so far, as well as providing an objective approach for the further development of these techniques. PMID- 19187990 TI - Against the grain: safeguarding rice from rice blast disease. AB - Rice is the staple diet of more than three billion people. Yields must double over the next 40 years if we are to sustain the nutritional needs of the ever expanding global population. Between 10% and 30% of the annual rice harvest is lost due to infection by the rice blast fungus Magnaporthe oryzae. Evaluation of genetic and virulence diversity of blast populations with diagnostic markers will aid disease management. We review the M. oryzae species-specific and cultivar specific avirulence determinants and evaluate efforts towards generating durable and broad-spectrum resistance in single resistant cultivars or mixtures. We consider modern usage of fungicides and plant defence activators, assess the usefulness of biological control and categorize current approaches towards blast tolerant genetically modified rice. PMID- 19187991 TI - Bioanode performance in bioelectrochemical systems: recent improvements and prospects. AB - In a bioelectrochemical system (BES) operated with a bioanode, the anode performance plays an important part in the overall performance. Fundamental aspects of bioanodes have been intensively investigated, enabling us to better understand the growth, kinetics functioning and interactions of anodophilic microorganisms. Recently, various technological advances have improved the properties and operation of anodes and have increased bioanode performance by up to tenfold. To further boost the performance of bioanodes by several orders of magnitude, practical microbiological approaches deserve more investigation. This article reviews the factors affecting bioanode performance, the recent advances and the prospective strategies for improving it. Future application perspectives of bioanodes are also proposed. PMID- 19187992 TI - Tipping the balance: modulating the Wnt pathway for tissue repair. AB - The Wnt signaling pathway has a crucial role in regulating cell growth and differentiation and is required for tissue homeostasis and repair. Although constitutive activation of the Wnt pathway can lead to abnormal cell growth and cancer, modulation of Wnt signaling might have a therapeutic benefit for tissue regeneration in numerous diseases. Recently, preclinical studies have demonstrated that treatments with antibodies against the Wnt inhibitor Dickkopf1 (DKK1) and with the positive Wnt modulator R-Spondin1 (RSpo1) were sufficient to repair the bone lesions in multiple myeloma and rheumatoid arthritis and to restore the damaged mucosa in experimental colitis, respectively. A remarkable balance is set for Wnt signaling by secreted proteins such as RSpo1 and DKK1, which help to regulate tissue homeostasis. As physiological Wnt response is essential for the regeneration of damaged tissues, modulation of the Wnt pathway might be beneficial for the treatment of multiple human diseases. PMID- 19187993 TI - Dose escalation model for limited-stage small-cell lung cancer. AB - BACKGROUND AND PURPOSE: To construct a model for the local dose-response relationship of limited-stage small-cell lung cancer (LS-SCLC) along with subsequent models for dose-per-fraction escalation. MATERIALS AND METHODS: Reported doses and 2-year progression free survival (PFS(2year)) rates from LS SCLC studies were used to construct a dose-response model. Modeling incorporated effects of chemotherapy, incomplete repair of sublethal damage and variations in both kick-off time for rapid reproliferation and effective tumor cell doubling time. Response was modeled as a function of overall treatment duration in order to shed light on the optimum radiotherapy treatment duration. Subsequent models were constructed for more aggressive radiotherapy by means of dose-per-fraction escalation for once a day (QD) and twice a day (BID) fractionation schedules. RESULTS: Modeling response as a function of treatment duration resulted in an optimum treatment duration of 3 weeks for both fractionation schedules. Modeling for dose-per-fraction escalation resulted in an increase in the expected local PFS(2year) for both fractionation schedules with possible gains in a BID schedule with lower expected late complication rates than a QD schedule yielding the same expected local PFS(2year). CONCLUSIONS: This modeling suggests more aggressive treatment through escalation of dose-per-fraction, which may result in substantial gains in local PFS(2year). This points towards treatment durations of 3 weeks as being optimal with a BID schedule predicting possible gains in local PFS(2year) at lower rates of expected late complications than a QD schedule. PMID- 19187994 TI - Influence of pH, salt, and temperature on pressure inactivation of hepatitis A virus. AB - The effects of pH (3-7), NaCl (0-6%), and temperature on pressure inactivation of hepatitis A virus (HAV) were determined. The HAV samples were treated at 400 MPa for 1 min at 5, 20, and 50 degrees C. Decreasing solution pH enhanced pressure inactivation of HAV. This enhanced inactivation effect was most evident at 20 degrees C. A baroprotective effect was observed for NaCl concentrations from 1 to 6%. For example, a treatment of 400 MegaPascals (MPa) for 1 min at 50 degrees C reduced the HAV titers by 4.0, 4.1, 1.3 and 0.4 log plaque forming units (PFU)/ml for NaCl concentrations of 0, 1, 3, and 6%, respectively. Overall, increasing the treatment temperature enhanced pressure inactivation of HAV in the solutions. The pressure resistance of HAV in oysters was also examined. Temperature in the range of 5 to 50 degrees C did not significantly affect the pressure inactivation of HAV within oyster homogenates. It is concluded that HPP treatment of oysters at temperatures above room temperature would not provide any additional benefit for inactivation of HAV. However, the observation that HAV inactivation is enhanced in acidic matrices is information that may be useful for designing product formulations and processing parameters for high pressure processing of products such as low pH fruit juices and salsa. PMID- 19187995 TI - A qualified presumption of safety approach for the safety assessment of Grana Padano whey starters. AB - A Qualified Presumption of Safety (QPS) approach was applied to dominant lactic acid bacteria (LAB) associated with Grana Padano cheese whey starters. Thirty-two strains belonging to Lactobacillus helveticus, Lactobacillus delbrueckii subsp. lactis, Streptococcus thermophilus, and Lactobacillus fermentum, and representing the overall genotypic LAB diversity associated with 24 previously collected whey starters [Rossetti, L., Fornasari, M.E., Gatti, M., Lazzi, C., Neviani, E., Giraffa, G., 2008. Grana Padano cheese whey starters: microbial composition and strain distribution. International Journal of Food Microbiology 127, 168-171], were analyzed. All L. helveticus, L. delbrueckii subsp. lactis, and S. thermophilus isolates were susceptible to four (i.e. vancomycin, gentamicin, tetracycline, and erythromycin) of the clinically most relevant antibiotics. One L. fermentum strain displayed phenotypic resistance to tetracycline (Tet(R)), with MIC of 32 microg/ml, and gentamycin (Gm(R)), with MIC of 32 microg/ml. PCR was applied to this strain to test the presence of genes tet(L), tet(M), tet(S), and aac(6')-aph(2')-Ia, which are involved in horizontal transfer of Tet(R) and Gm(R), respectively but no detectable amplification products were observed. According to QPS criteria, we conclude that Grana cheese whey starters do not present particular safety concerns. PMID- 19187996 TI - Broth and agar hop-gradient plates used to evaluate the beer-spoilage potential of Lactobacillus and Pediococcus isolates. AB - Identification of the beer-spoilage Lactobacillus and Pediococcus bacteria has largely taken two approaches; identification of spoilage-associated genes or identification of specific species of bacteria regardless of ability to grow in beer. The problem with these two approaches is that they are either overly inclusive (i.e., detect all bacteria of a given species regardless of spoilage potential) or overly selective (i.e., rely upon individual, putative spoilage associated genes). Our goal was to design a method to assess the ability of Lactobacillus and Pediococcus to spoil beer that is independent of speciation or genetic background. In searching for a method by which to differentiate between beer-spoilage bacteria and bacteria that cannot grow in beer, we explored the ability of lactobacilli and pediococci isolates to grow in the presence of varying concentrations of hop-compounds and ethanol in broth medium versus on agar medium. The best method for differentiating between bacteria that can grow in beer and bacteria that do not pose a threat as beer-spoilage organisms was found to be a hop-gradient agar plate containing ethanol. This hop-gradient agar plate technique provides a rapid and simple solution to the dilemma of assessing the ability of Lactobacillus and Pediococcus isolates to grow in beer, and provides new insights into the different strategies used by these bacteria to survive under the stringent conditions of beer. PMID- 19187997 TI - 1,5-Anhydroglucitol reflects postprandial hyperglycemia and a decreased insulinogenic index, even in subjects with prediabetes and well-controlled type 2 diabetes. AB - To examine the serum 1,5-anhydroglucitol (AG) levels as a surrogate measure of postprandial hyperglycemia (PPH) and insulin secretion in a wide range of hyperglycemia, we compared the relationship between the glycemic index during a 75g oral glucose tolerance test (OGTT) and the insulinogenic index and 1,5-AG according the overall glycemic state. Fasting serum 1,5-AG levels were lower in the type 2 diabetic group (18.0+/-7.0microg/mL) than in the normal glucose tolerance (NGT, 25.4+/-4.0microg/mL), impaired fasting glucose (IFG, 24.6+/ 6.2microg/mL), and impaired glucose tolerance (IGT, 22.1+/-6.2microg/mL) groups and were clearly correlated with glycemic values from the OGTT. 120-min post challenge plasma glucose (PPG(120)) emerged as an independent predictor for 1,5 AG levels after multiple linear regression analysis (beta=-0.554, P<0.001). Additionally, 1,5-AG levels were significantly correlated with PPG(120) in each quartile of A1C, and the coefficients increased with higher A1C quartiles. Subjects with low 1,5-AG levels had both increased insulin resistance and decreased insulin secretion. Decreased 1,5-AG levels are closely correlated with PPH and decreased insulin secretion capacity across a wide range of glycemia, even in relatively well-controlled diabetes. PMID- 19187998 TI - The usefulness and cost of a shorter duration of fasting in workplace screening for glucose intolerance and metabolic syndrome. AB - For worksite biometric screening, we shortened traditional fasting instructions to 4h. The positive predictive value (PPV) of elevated capillary glucose test results (> or =100mg/dL) for predicting elevated values obtained on repeat testing after an 8h fast was 57%, which rivals the PPV of an 8h fasting glucose. PMID- 19187999 TI - Relation of Hemoglobin A1c to myocardial acoustic densitometry and left ventricular diastolic function in patients with Type 2 diabetes mellitus and without evident heart disease. AB - AIM: The aim of the study was to detect the echocardiographic sensitive indexes for prediction of the subclinical cardiac dysfunction and to evaluate the relation between Hemoglobin A1c (HbA1c) and myocardial acoustic densitometry as well as cardiac function. METHODS: Fifty DM2 patients (48.5+/-8.6 years) without evident heart disease and 50 age- and sex-matched normal controls (47.6+/-8.8 years) were enrolled. Conventional echocardiography, tissue Doppler imaging and acoustic densitometry of both groups were measured. HbA1c of DM2 patients were determined. RESULTS: There were no significant differences in systolic indexes between the two groups. Mean LV myocardial early diastolic velocity Em and Em/Am were lower in DM group than control group. Mean LV myocardial late diastolic velocity Am and E/Em were higher in DM group than control group. IVS-IBS% and LVPW-IBS% were higher in DM group than control group. IVS-CVIB and LVPW-CVIB were lower in DM group than control group. HbA1c was negatively correlated with E/A (gamma=-0.310, P<0.05), Em (gamma=-0.409, P<0.01), Em/Am (gamma=-0.380, P<0.01) and positively correlated with E/Em (gamma=0.488, P<0.01). Significant positive correlation was present between HbA1c and IVS-IBS% (gamma=0.679, P<0.01), LVPW IBS% (gamma=0.666, P<0.01). CONCLUSIONS: The diastolic dysfunction and abnormal myocardial acoustic densitometry exist before the systolic function damage in DM2 patients. Tissue Doppler imaging and ultrasonic integrated backscatter can be used as sensitive means for early assessing myocardial histological changes in DM2 patients. HbA1c is related with both diastolic dysfunction and acoustic densitometry. PMID- 19188000 TI - Severe hypoglycaemia during pregnancy in women with type 1 diabetes: possible role of renin-angiotensin system activity? AB - AIMS: To investigate whether increased risk of severe hypoglycaemia in early pregnancy is related to pregnancy-induced changes in renin-angiotensin system (RAS) activity in women with type 1 diabetes (T1DM). METHODS: Severe hypoglycaemic events the year preceding pregnancy were recorded retrospectively in 107 consecutive pregnant women with T1DM. Events during pregnancy were recorded prospectively. Measurements of ACE, renin and angiotensinogen were determined at 8, 14, 21, 27 and 33 weeks and postpartum. RESULTS: The rate of severe hypoglycaemia was 1.1 and 5.3 events/patient-year the year preceding pregnancy and in first trimester, respectively (p<0.0001). Levels of ACE, renin or angiotensinogen did not differ between women with and without severe hypoglycaemia during pregnancy. Multivariate regression analysis identified a positive association between rate of severe hypoglycaemia the year preceding pregnancy and postpartum ACE activity (relative rate of severe hypoglycaemia above versus below median ACE activity: 4.4 (CI: 1.7-11.9), p=0.003). No association was found between severe hypoglycaemia during pregnancy and renin angiotensin system activity at 8 weeks. CONCLUSIONS: In early pregnancy increased RAS activity does not explain the 5-fold increase in severe hypoglycaemia in women with T1DM. A positive association between occurrence of severe hypoglycaemia and ACE activity outside pregnancy was demonstrated. PMID- 19188001 TI - Evaluation of institutional practices for prevention of phlebotomy-associated percutaneous injuries in hospital settings. AB - BACKGROUND: To reduce the incidence of phlebotomy-related percutaneous injuries (PIs), factors that contribute to these injuries must be identified. This study examined institutional phlebotomy practices, policies, perceptions, and culture to identify facilitators and barriers that appear to have the greatest impact in preventing injuries. METHODS: During site visits at study hospitals, observational data were collected during the performance of phlebotomy. In addition, interviews and focus groups were conducted with hospital personnel involved in phlebotomy procedures. RESULTS: Nine hospitals participated in the study. A total of 126 phlebotomy procedures were observed. Health care personnel chose devices with safety features for the majority of observed procedures (n = 122, 97%). Recommended phlebotomy practices for handling needles after use were observed in 42% to 92% of procedures. Adherence varied by type of device, occupation, and facility PI rate. In the 23 interviews and 9 focus groups, participants identified factors that facilitated PI prevention such as the availability and use of devices with safety mechanisms, adherence to recommended safe needle-handling practices, and institutional phlebotomy training. CONCLUSION: The quantitative and qualitative data indicate that a wide array of factors can affect phlebotomy-related practices and perceptions. Prevention of PIs may require comprehensive, multifaceted intervention efforts to improve the safety culture and reduce PIs and exposure to bloodborne pathogens in health care facilities. PMID- 19188002 TI - Surgical hand rub: Influence of duration of application on the immediate and 3 hours effects of n-propanol and isopropanol. AB - BACKGROUND: The recommended duration for surgical hand treatment has been changed from 10 over 5 to 3 minutes and even shorter. OBJECTIVES: Our objective was to study the impact of the length of surgical hand antisepsis with n-propanol 60% (vol/vol) or isopropanol 70% (vol/vol) applied for 1, 3, or 5 minutes on the reduction of resident hand flora in the setting of the microbiologic laboratory for experimental and applied testing of disinfectants and antiseptics at the Medical University Vienna, Austria, using a Latin Square design. METHODS: Our methods were according to the Austrian Guidelines for Testing Products for Surgical Hand Antisepsis. The release of bacterial hand flora of 21 subjects is assessed before and immediately after disinfection from one hand and 3 hours later from the other, meanwhile gloved, hand. Mean reduction factors (RF) are calculated. RESULTS: The immediate mean log(10) RFs with n-propanol or isopropanol were 1.05, 2.03, and 2.30 and 0.74, 1.48, and 2.12, respectively, when applied for 1, 3, or 5 minutes, respectively. After 3 hours, the respective mean log(10) RFs were 0.45, 1.01, and 1.60 and 0.19, 0.79, and 1.03. Thus, with increasing length of application, a highly significant trend (P < .001) toward higher log(10) reductions was demonstrated. At both sampling times, n-propanol was more effective than isopropanol at the corresponding treatments. Furthermore, a highly significant (P < .001) association was found between the individual volunteers and the effect of the antiseptics on their hands. CONCLUSION: The efficacy of surgical antisepsis is significantly associated with the length of application. PMID- 19188003 TI - Evaluation of the filtration performance of 21 N95 filtering face piece respirators after prolonged storage. AB - BACKGROUND: Organizations are stockpiling respirators to prepare for an influenza pandemic. To understand better the effects of prolonged storage, this investigation evaluated the filtration efficiency of 21 different models of National Institute for Occupational Safety and Health (NIOSH)-certified disposable N95 filtering face piece respirators. These respirators had been stored in their original packaging for a period of at least 6 years in research laboratories and dry warehouse facilities, ranging in temperature between 15 degrees C and 32 degrees C and relative humidity between 20% and 80%. METHODS: Filter penetration was measured using an abbreviated version of the NIOSH respirator certification test incorporating a polydisperse sodium chloride aerosol at 85 L/min. RESULTS: Of the 21 respirator models tested, 19 models had both average penetration results of less than 5%. Mean initial penetration values ranged from 0.39% to 5.83%, whereas mean maximum penetration values ranged from 0.95% to 5.83%. There did not appear to be any correlation between the length of storage and failure to pass the filtration test. CONCLUSION: Results indicate that most N95 filtering face piece respirators stored for up to 10 years at warehouse conditions will likely have expected levels of filtration performance and that the degree of filtration efficiency degradation is likely model specific. PMID- 19188004 TI - Vancomycin-resistant Enterococcus colonization before admission to the intensive care unit: a clinico-epidemiologic analysis. AB - BACKGROUND: Asymptomatic vancomycin-resistant Enterococcus (VRE) colonization is known to precede actual infection. Since VRE was first isolated in Korea in 1992, the VRE isolation rate has rapidly increased in tertiary hospitals. METHODS: We performed a prospective observational study to estimate the prevalence of VRE colonization among inpatients at the time of intensive care unit (ICU) admission. From March through December 2007, rectal swab cultures were taken in all patients admitted to the ICU. We aimed to identify the risk factors for VRE colonization on admission. RESULTS: During the study period, 34 (4.4%) out of 780 patients were already colonized with VRE before ICU admission: 21 out of 323 patients from general wards (6.5%) and 13 out of 437 patients from outside the hospital (2.97%). VRE-colonized patients were more likely than uncolonized patients to have infectious diseases and to have been referred from outside hospitals (P < .01). Previous hospitalization (P = .01) and antibiotic exposure (P < .01) were risk factors for VRE colonization before ICU admission. Pulsed-field gel electrophoresis patterns were diverse. Initial VRE colonization correlated to poor prognosis. CONCLUSION: VRE colonization rate was not negligible among newly admitted ICU patients, suggesting that an active surveillance program focusing on high-risk groups may be helpful. PMID- 19188005 TI - Functional brain changes in early Parkinson's disease during motor response and motor inhibition. AB - Motor impairment represents the main clinical feature of Parkinson's disease (PD). Cognitive deficits are also frequently observed in patients with PD, with a prominent involvement of executive functions and visuo-spatial abilities. We used event-related functional MRI (fMRI) and a paradigm based on visual attention and motor inhibition (Go/NoGO-task) to investigate brain activations in 13 patients with early PD in comparison with 11 healthy controls. The two groups did not report behavioural differences in task performance. During motor inhibition (NoGO effect), PD patients compared to controls showed an increased activation in the prefrontal cortex and in the basal ganglia. They also showed a reduced and less coherent hemodynamic response in the occipital cortex. These results indicate that specific cortico-subcortical functional changes, involving not only the fronto-striatal network but also the temporal-occipital cortex, are already present in patients with early PD and no clinical evidence of cognitive impairment. We discuss our findings in terms of compensatory mechanisms (fronto striatal changes) and preclinical signs of visuo-perceptual deficits and visual hallucinations. PMID- 19188006 TI - Root uptake of radionuclides following their acute soil depositions during the growth of selected food crops. AB - Greenhouse experiments were performed to investigate the root uptake of radionuclides following their acute soil deposition during the growth of several food crops. For this purpose, the soil under the standing plants was contaminated without any direct contamination of their stems or leaves. The intention of this design was to differentiate foilar uptake and root uptake subsequent to a radionuclide deposition during the vegetation period. Soil-to-plant transfer of a radionuclide was quantified with its aggregated transfer factors specified for the time periods from deposition until harvest (T(ag)(a), m(2)kg(-1)). Deposition time-dependent T(ag)(a) values of Mn, Co, Sr and Cs for selected crop species were measured in an acid sandy soil. For rice and Chinese cabbage, HTO experiments were also carried out using this soil. Particularly for rice, experiments with various paddy soils were also performed for (90)Sr and (137)Cs. The obtained T(ag)(a) values varied considerably with the radionuclides, plant species, and times of deposition. Recommendations about, and limitations in, the use of the T(ag)(a) values were discussed. PMID- 19188007 TI - Re: Lemierre's syndrome: a forgotten disease. PMID- 19188008 TI - Effect of 3 years' free-air exposure to elevated ozone on mature Norway spruce (Picea abies (L.) Karst.) needle epicuticular wax physicochemical characteristics. AB - We examined the effect of ozone (O(3)) on Norway spruce (Picea abies) needle epicuticular wax over three seasons at the Kranzberg Ozone Fumigation Experiment. Exposure to 2x ambient O(3) ranged from 64.5 to 74.2 microl O(3) l(-1) h AOT40, and 117.1 to 123.2 nl O(3) l(-1) 4th highest daily maximum 8-h average O(3) concentration. The proportion of current-year needle surface covered by wax tubes, tube aggregates, and plates decreased (P=0.011) under 2x O(3). Epistomatal chambers had increased deposits of amorphous wax. Proportion of secondary alcohols varied due to year (P=0.004) and O(3) treatment (P=0.029). Secondary alcohols were reduced by 9.1% under 2x O(3). Exposure to 2x O(3) increased (P=0.037) proportions of fatty acids by 29%. Opposing trends in secondary alcohols and fatty acids indicate a direct action of O(3) on wax biosynthesis. These results demonstrate O(3)-induced changes in biologically important needle surface characteristics of 50-year-old field-grown trees. PMID- 19188009 TI - Identification of Chinese cabbage genotypes with low cadmium accumulation for food safety. AB - The pot-culture experiment and field studies were conducted to screen out and identify cadmium (Cd) excluders from 40 Chinese cabbage genotypes for food safety. The results of the pot-culture experiment indicated that the shoot Cd concentrations under three treatments (1.0, 2.5 and 5.0 mg Cd kg(-1) Soil) varied significantly (p<0.05), with average values of 0.70, 3.07 and 5.83 mg kg(-1), respectively. The Cd concentrations in 12 cabbage genotypes were lower than 0.50 mg kg(-1). The enrichment factors (EFs) and translocation factors (TFs) in 8 cabbage genotypes were lower than 1.0. The field studies further identified Lvxing 70 as a Cd-excluder genotype (CEG), which is suitable to be planted in low Cd-contaminated soils (Cd concentration should be lower than 1.25 mg kg(-1)) for food safety. PMID- 19188010 TI - Doorways in nature: syndemics, zoonotics, and public health. A commentary on Rock, Buntain, Hatfield & Hallgrimsson. PMID- 19188011 TI - The structuring of occupational stressors in a Post-Fordist work environment. Moving beyond traditional accounts of demand, control and support. AB - In this study, the traditional components of the Demand-Control-Support-Model are extended with a broad number of occupational stressors, while investigating associations with persistent fatigue, musculoskeletal complaints and emotional well-being. Furthermore, it is assessed how these stressors are structured into distinct dimensions within a population of wage-earners. In a representative cross-sectional sample of 11,099 Flemish (Belgian) employees a range of loglinear techniques is used: logit modelling, latent class analysis and Modified LISREL modelling. Quantitative, emotional and physical demands, repetitive movements, types of work schedules, problematic autonomy, task variation, social relationships with superiors, job insecurity and bullying behaviour are associated with at least one of the health outcomes, while sudden schedule changes are not. These occupational stressors constitute five dimensions: immaterial demands, physical demands, control over the work environment, social relationships at work and employment uncertainty. These latent dimensions are all significantly related with at least one of the health outcomes - with immaterial demands having the strongest effects. Contemporary work is characterised by a complex combination of stressors, structured within the population into a number of dimensions. More research on the interrelatedness of occupational stressors is needed. PMID- 19188012 TI - Modelling the effects of intimate partner violence and access to resources on women's health in the early years after leaving an abusive partner. AB - Although the negative health effects of intimate partner violence (IPV) are well documented, little is known about the mechanisms or determinants of health outcomes for women who had left their abusive partners. Using data collected from a community sample of 309 Canadian women who left an abusive partner, we examined whether women's personal, social and economic resources mediate the relationships between the severity of past IPV and current health using structural equation modelling. A good fit was found between the model and data for hypothesized models of mental and physical health. In the mental health model, both the direct and total indirect effects of IPV were significant. In the physical health model, the direct effect of IPV on physical health was about four times as large as the total indirect effects. In both models, more severe past IPV was associated with lower health and women's personal, social, and economic resources, when combined, mediated the relationship between IPV and health. These findings demonstrate that the health outcomes of IPV for women who have left an abusive partner must be understood in context of women's resources. PMID- 19188013 TI - Resistance to degradation of resin-modified glass-ionomer cements dentine bonds. AB - OBJECTIVE: To evaluate the effect of EDTA pre-treatment of dentine on resistance to degradation of the bond between dentine and resin-modified glass-ionomer cements. METHODS: Sixty non-carious human molars underwent cavity preparations. Teeth were restored with Fuji II LC or Vitremer. Half of the cavities were restored following manufacturers' instructions whereas the other half was pre treated with EDTA (0.1M, pH 7.4) for 60s. Teeth were stored in water at 37 degrees C for 24h, 3 months or submitted to 10% NaOCl immersion for 5h. Teeth were sectioned into beams (1+/-0.1mm) and tested to failure in tension at 0.5mm/min. Bond strength data (MPa) were analyzed by ANOVA and SNK multiple comparisons tests (p<0.05). RESULTS: When EDTA was used for pre-treatment of dentine, higher bond strengths were observed for both cements. Degradation challenges produced a decrease in bond strength values only in the Vitremer group. This decrease was avoided when EDTA was used for dentine treatment before restoring with Vitremer. CONCLUSIONS: EDTA pre-treatment of dentine increases bond strength of resin modified glass-ionomers cements to dentine and improves resistance to degradation of the bond between Vitremer and dentine. PMID- 19188014 TI - Enhanced biodegradation of pentachlorophenol in unsaturated soil using reversed field electrokinetics. AB - This study investigated the use of electrokinetics in unsaturated soil to promote biodegradation of pentachlorophenol through increased contact between bacteria and contaminant. Soil microcosms, contaminated with approximately 100 mg kg(-1) pentachlorophenol (containing [(14)C]-PCP as a tracer), and inoculated with a specific pentachlorophenol-degrading bacterium (Sphingobium sp. UG30-1 x 10(8) cfu g(-1)) were subjected to constant and regularly reversed electric currents (10 mA). The former caused large pH and moisture content changes due to water electrolysis and electroosmotic effects, with subsequent negative impacts on biodegradation parameters including enzyme activity and contaminant mineralisation (as measured by (14)CO(2) evolution rate). The reversed field caused little change in pH and moisture content and led to more rapid contaminant mineralisation, lower soil contaminant concentration in the majority of the microcosms and increased soil enzyme activity (with the exception of soil immediately adjacent to the anode). The presence of an electric field, if suitably applied, may therefore enhance contaminant biodegradation in unsaturated soil. PMID- 19188015 TI - Systemic presence and tumor-growth promoting effect of ovarian carcinoma released exosomes. AB - Exosomes are membrane vesicles that are released from many different cell types. Tumor derived-exosomes play a role in immune suppression. We hypothesized that in ovarian carcinoma patients exosomes initially produced at the local abdominal site may become systemic. We examined paired samples of ascites and blood from ovarian carcinoma patients for the presence of exosomes. We also studied the requirements for exosomal uptake by immune cells, the role of phosphatidyl-serine (PS) as uptake signal and the effect of exosome application on tumor growth. We used exosomes from ovarian carcinoma cell lines, malignant ascites and sera from ovarian carcinoma patients isolated by ultracentrifugation. PS-displayed by exosomes was detected by Anexin-V-FITC staining of latex beads adsorbed exosomes. For uptake experiments, labeled exosomes were exposed to cells in the presence or absence of cold Annexin-V as competitor. Uptake was examined by fluorescent microscopy and cytofluorographic analysis. Effects of exosomes on tumor growth were studied using SKOV3ip ovarian carcinoma cells in CD1 nu/nu mice. We found that malignant ascites-derived exosomes cargo tumor progression related proteins such as L1CAM, CD24, ADAM10, and EMMPRIN. We observed that exosomes become systemic via the blood stream. Uptake of ovarian carcinoma exosomes by NK cells was found to require PS at the exosomal surface but the presence of PS was not sufficient. Application of malignant ascites-derived exosomes to tumor bearing mice resulted in augmented tumor growth. Exosomes from the serum of tumor patients could be isolated from only one ml of blood and this analysis could serve for diagnostic purposes. We propose that tumor-derived exosomes could play a role in tumor progression. PMID- 19188016 TI - Effect of Ganoderma on drug-sensitive and multidrug-resistant small-cell lung carcinoma cells. AB - Multidrug resistance is a major problem in small-cell lung cancer (SCLC). Ganoderma lucidum is a widely used herb in traditional Chinese medicine. We tested the effects of Ganoderma on drug-sensitive (H69) and multi-drug resistant (VPA) human SCLC cells. Both cells showed equal cytotoxicity when incubated with extracts of mycelia of 9 species of Ganoderma, including G. lucidum. Cells treated with the IC(50) of cytotoxic Ganoderma and analyzed by flow cytometry-PI staining showed increases in S phase. When compared untreated controls or SCLC cells treated with extracts of non-cytotoxic Ganoderma species, cells treated with extracts of cytotoxic Ganoderma species responded with an induction of apoptosis similar to cells treated with the chemotherapeutic drugs etoposide and doxorubicin. This was shown by four criteria: increased DNA fragmentation within cells as measured by ELISA; increased TUNEL staining for DNA breaks; increased specific activities of caspases 3 and 9, but not caspase 8 by colorimetric assays, indicating the endogenous pathway; and similar patterns changes in the expressions of 9 genes involved in the cell cycle/apoptosis, as measured by RT PCR and capillary electrophoresis. Pre-incubation of drug-resistant SCLC cells with cytotoxic Ganoderma reduced the IC(50) for etoposide (3.4-0.21 microM) and doxorubicin (0.19-0.04 microM). These results show that extracts of several species of Ganoderma are cytotoxic to both drug-sensitive and drug-resistant SCLC cells, are pro-apoptotic, induce gene expression patterns that are similar to SCLC cells treated with chemotherapeutic drugs, and can reverse resistance to chemotherapeutic drugs. PMID- 19188017 TI - Removal of basic dye using raw and acid activated bentonite samples. AB - The adsorption behavior of crystal violet (CV(+)) from aqueous solution onto raw (RB) and acid activated (AAB) bentonite samples was investigated as a function of parameters such as initial CV(+) concentration, contact time and temperature. The Langmuir and Freundlich adsorption models were applied to describe the equilibrium isotherms. The adsorption rate was fast and more than half of the adsorbed-CV(+) was removed in the first 55 min for RB and 25 min for 0.2-AAB at the room temperature (C(0)=1.2 mmol/g). The pseudo-first order, and pseudo-second order kinetic models were used to describe the kinetic data and rate constants were evaluated. The results best fit the pseudo-second order kinetic model with the rate constant, k(2), in the range of 13.52 x 10(-2) to 5.53 x 10(-2)g/mmol and 52.70 x 10(-2) to 6.69 x 10(-2)mmol/g min for RB and 0.2-AAB sample, respectively (C(0)=0.1-1.2 mmol/g). PMID- 19188018 TI - Competitive biosorption of Pb2+, Cu2+ and Zn2+ ions from aqueous solutions onto valonia tannin resin. AB - Competitive biosorption of Pb(2+), Cu(2+) and Zn(2+) on valonia tannin resin (VTR) present in binary and ternary mixture were compared with the single metal solution. The effects of the presence of one metal ion on the biosorption of the other metal ion were investigated in terms of equilibrium isotherm and biosorption yield. Experimental results indicated that the uptake capacity and biosorption yield of one metal ion were reduced by the presence of the other metal ion. The extent of adsorption capacity of the binary and ternary metal ions tested on VTR was low (18-80%) as compared to single metal ions. Comparisons between biosorption of Pb(2+), Cu(2+) and Zn(2+) ions by the biomass of VTR in the binary and ternary solution could lead to the conclusion that biosorption of Pb(2+) ions was preferential to that of Cu(2+) and Zn(2+) ions. VTR removed the target metal ions in the selectivity order of Pb(2+)>Cu(2+)>Zn(2+). The biosorption equilibrium data for the single metal solutions fitted the Langmuir model well. Results show that the pseudo-second-order equation provides the best correlation for the biosorption process in the single-ion system. PMID- 19188019 TI - Electrochemical oxidation of model compounds and olive mill wastewater over DSA electrodes: 1. The case of Ti/IrO(2) anode. AB - The electrochemical oxidation of olive mill wastewater (OMW) and model compounds over a Ti/IrO(2) anode was studied by means of cyclic voltammetry and bulk electrolysis. Experiments were conducted at 1300 mg/L initial COD, 0-1.23V vs SHE and 1.4-1.54V vs SHE potential windows, 50 mA/cm(2) current density, 0-25 mM NaCl, 60-80 degrees C temperature and acidic conditions. The reactivity of model compounds decreases in the order phenol approximately p-coumaric acid>cinnamic acid>caffeic acid. Partial and total oxidation reactions occur with the overall rate following zero-order kinetics with respect to COD and increasing with temperature. Oxidation of OMW at 43 Ah/L, 80 degrees C and in the presence of 5mM NaCl leads to complete color and phenols removal, elimination of ecotoxicity but moderate (30%) COD reduction. Similar performance can be achieved at 6 Ah/L in the presence of 15 mM NaCl. In the absence of salt, the respective color and phenols removal (at 6 Ah/L) is less than 10%. Excessive salinity (25 mM), although does not change color, phenols and COD removal, has an adverse effect on ecotoxicity. PMID- 19188020 TI - Risk reduction in road and rail LPG transportation by passive fire protection. AB - The potential reduction of risk in LPG (Liquefied Petroleum Gas) road transport due to the adoption of passive fire protections was investigated. Experimental data available for small scale vessels fully engulfed by a fire were extended to real scale road and rail tankers through a finite elements model. The results of mathematical simulations of real scale fire engulfment scenarios that may follow accidents involving LPG tankers proved the effectiveness of the thermal protections in preventing the "fired" BLEVE (Boiling Liquid Expanding Vapour Explosion) scenario. The presence of a thermal coating greatly increases the "time to failure", providing a time lapse that in the European experience may be considered sufficient to allow the start of effective mitigation actions by fire brigades. The results obtained were used to calculate the expected reduction of individual and societal risk due to LPG transportation in real case scenarios. The analysis confirmed that the introduction of passive fire protections turns out in a significant reduction of risk, up to an order of magnitude in the case of individual risk and of about 50% if the expectation value is considered. Thus, the adoption of passive fire protections, not compulsory in European regulations, may be an effective technical measure for risk reduction, and may contribute to achieve the control of "major accidents hazards" cited by the European legislation. PMID- 19188021 TI - Reliable prediction of electric spark sensitivity of nitramines: a general correlation with detonation pressure. AB - For nitramines, a general correlation has been introduced to predict electric spark sensitivity through detonation pressure. New method uses maximum obtainable detonation pressure as a fundamental relation so that it can be corrected for some nitramines which have some specific molecular structure. There is no need to use crystal density and heat of formation of nitramine explosives for predicting detonation pressure and electric spark sensitivity. The predicted electric spark sensitivities are compared with calculated results on the basis of quantum mechanical computations for some nitramines that latter can be applied. The root mean square (rms) deviations from experiment for new method and the predicted results of complicated quantum mechanical method are 1.18 and 3.49J, respectively. PMID- 19188022 TI - Analyses of phosphorus in sewage by fraction method. AB - Several forms of the phosphorus in wastewater were analyzed using a fraction method with a membrane reactor. A primary aim of this study was to differentiate the two main groups, as either "biologically bound" or "physicochemically bound," with the quantification of bound phosphorus in animated sludge. After the four level extraction treatment with distilled water and various solvents such as bicarbonate-dithionate, NaOH and HCl, the redox-sensitive-P and P bound to Fe(III)-hydroxides were found as main precipitation products in the anaerobic chamber (AN). Whereas the aerobic chamber (AE) contained mainly the precipitates of P bound to metals, such as Al and Fe, sorptive-bound phosphorus on metals and organic compounds etc. With iron precipitation, the levels of Fe-P, Al-P, and Fe P adsorbed by Fe(OH)(3) were particularly high in the AE and AN. The sum of inorganic-P (physicochemically bound phosphorus) increased with iron precipitation, whereas that of organic-P and poly P (biologically bound phosphorus) decreased. PMID- 19188023 TI - Heavy metals retention capacity of a non-conventional sorbent developed from a mixture of industrial and agricultural wastes. AB - Zinc and copper removal from aqueous solutions using brine sediments (industrial residue), sawdust (agricultural residue) and the mixture of both materials has been researched through batch and column tests. Brine sediments were found to be mainly constituted by halite and calcite, while its main cations exchangeable were sodium, calcium, magnesium and potassium. In sawdust the main exchangeable cations detected were calcium, magnesium, sodium and potassium. FT-IR spectra of sawdust and brine sediment-sawdust mixture showed that brine sediments produced important changes in carboxylic, alcoholic and phenolic groups present in the sawdust. The maximum zinc adsorption capacity was found to be 4.85, 2.58 and 5.59 mg/g using an adsorbent/solution ratio of 1/40, for brine sediments, sawdust and the mixture, respectively. For copper, the maximum adsorption capacity was found to be 4.69, 2.31 and 4.33 mg/g, using adsorbent/solution ratios of 1/40, for brine sediments, sawdust and the mixture, respectively. Maximum copper adsorption capacity of the mixture, on the contrary to zinc adsorption, was lightly inferior to maximum adsorption capacity obtained in brine sediments. Adsorption isotherms data adjusted better to the Langmuir model. Additionally, columns reached the saturation point at 690 min for zinc and 360 min for copper. The main mechanism involved in the removal of both metals may be the ionic exchange between sodium and calcium ions present in brine sediments and H(+) present in functional groups of sawdust. The use of brine sediments, sawdust and their mixture, presents an interesting option both, for wastewater decontamination (as a possible non conventional sorbent for the removal of heavy metals) and as a waste recycling option. PMID- 19188024 TI - Photodegradation of C.I. Reactive Red 2 in UV/TiO2-based systems: effects of ultrasound irradiation. AB - This investigation elucidated the decolorization of C.I. Reactive Red 2 (RR2) in US/TiO(2), UV/TiO(2) and UV/US/TiO(2) systems and evaluated the effect of ultrasound (US) irradiation in photocatalysis. The effects of RR2 concentration, temperature and the addition of NaCl, Na(2)S(2)O(8) and radical scavenger were determined. The decolorization reactions obeyed the pseudo-first-order kinetics in all tested systems. In US-related systems, the decolorization rate of RR2 declines as RR2 concentration increases. At pH 7, the decolorization rates followed the order UV/US/TiO(2) (0.94 h(-1))>UV/TiO(2) (0.85 h(-1))>US/TiO(2) (0.25 h(-1)). The promotion efficiencies of adding NaCl in US/TiO(2), UV/TiO(2) and UV/US/TiO(2) systems were 16%, 18% and 29%, respectively. The decolorization rate increased with the temperature; additionally, the decolorization rate in UV/US/TiO(2)/Na(2)S(2)O(8) exceeded that in UV/US/TiO(2). The inhibition of RR2 decolorization by adding 1-butanol reveals that the primary decolorization pathway involves hydroxyl radicals, and that direct oxidation by photogenerated holes is probably important in the UV/TiO(2)-based system. After 120 min of the reaction, the TOC degradation efficiencies of UV/TiO(2) and UV/US/TiO(2) systems were 47% and 63%, respectively. PMID- 19188025 TI - Optimization of Cu(II) biosorption onto Ascophyllum nodosum by factorial design methodology. AB - A Box-Behnken factorial design coupled with surface response methodology was used to evaluate the effects of temperature, pH and initial concentration in the Cu(II) sorption process onto the marine macro-algae Ascophyllum nodosum. The effect of the operating variables on metal uptake capacity was studied in a batch system and a mathematical model showing the influence of each variable and their interactions was obtained. Study ranges were 10-40 degrees C for temperature, 3.0 5.0 for pH and 50-150 mg L(-1) for initial Cu(II) concentration. Within these ranges, the biosorption capacity is slightly dependent on temperature but markedly increases with pH and initial concentration of Cu(II). The uptake capacities predicted by the model are in good agreement with the experimental values. Maximum biosorption capacity of Cu(II) by A. nodosum is 70 mg g(-1) and corresponds to the following values of those variables: temperature=40 degrees C, pH=5.0 and initial Cu(II) concentration=150 mg L(-1). PMID- 19188026 TI - Studies on the adsorption behavior of trace amounts of 90Sr2+, 140La3+, 60Co2+, Ni2+ and Zr4+ cations on synthesized inorganic ion exchangers. AB - Three inorganic ion exchangers namely potassium zinc hexacyanoferrate(II) (PZF), magnesium oxide-polyacrylonitrile composite (MgO-PAN) and ammonium molybdophosphate (AMP) were synthesized. The physicochemical properties of these ion exchangers were determined using different techniques including inductively coupled plasma (ICP), CHNSO elemental analysis, infrared spectroscopy (IR), X-ray diffraction (XRD), thermogravimetric (TGA) and pH - titration curve analysis. The solubility of the synthesized ion exchangers in different acidic and alkaline media, their thermal stability and the effect of gamma irradiation were investigated. It was observed that the exchange capacity of the ion exchangers depend upon the pH value of the solution used. Furthermore, the adsorption of (90)Sr(2+), (140)La(3+), (60)Co(2+) and the distribution coefficient of these ion exchangers for Ni(2+)and Zr(4+) were studied. The effect of parameters such as pH and contact time on the adsorption was also investigated and the optimum conditions for separation of these ions were determined. PMID- 19188027 TI - Altered glucose tolerance in women with deliberate self-harm. AB - Disturbances in glucose metabolism are of importance for violent behaviour in men, but studies in women are lacking. We used the 5h-oral glucose tolerance test (OGTT) in this study of 17 female psychiatric patients, selected for violent behaviour directed against themselves (deliberate self-harm) and 17 healthy controls matched for age and BMI. Following OGTT, patients had higher glucose levels at 30 min (p=0.007) and increased glucagon area under the curve (p=0.011). Since a co-morbid eating disorder might affect results, we as a post-hoc analysis subgrouped the patients and found that the increased glucagon levels only were present in patients with an eating disorder. In contrast, those without an eating disorder showed a significantly lower p-glucose nadir (p=0.015) and unaltered glucagon levels compared to controls. There were no significant differences in insulin and C-peptide levels between patients and controls. We conclude that deliberate self-harm in women may be associated with alterations in carbohydrate metabolism in certain groups. Eating disorder is a confounding factor. PMID- 19188028 TI - Can moderate elevations of parathyroid hormone acutely increase risk for ischemic cardiac arrhythmias? AB - There is suggestive evidence that chronic elevations of parathyroid hormone (PTH), associated with poor vitamin D status or low calcium intake, can increase risk for insulin resistance, weight gain, hypertension, and left ventricular hypertrophy, while stimulating production of acute phase reactants. New evidence that elevated PTH is prognostic for increased vascular mortality in very elderly subjects, prompts an examination of the possible impact of PTH on risk for arrhythmias. The cardiac effects of PTH are mediated by G protein-coupled receptors that activate phospholipase C (PLC). Catecholamines, angiotensin II, and endothelin have been shown to be arrhythmogenic for ischemic myocardium in animal studies; the receptors mediating this effect are all likewise linked to activation of PLC. Thus, it is reasonable to presume that a sufficient concentration of PTH can be arrhythmogenic in the ischemic heart. The extent to which this effect can be evoked by the high-normal PTH levels prevalent in many elderly subjects, can be assessed in epidemiological studies. PMID- 19188029 TI - Rh factor, family history and risk of breast cancer: a case-control study in Uruguay. AB - OBJECTIVE: To explore possible relationships among blood factors, family history of breast cancer (BC) and the risk of the disease, a case-control study was carried out in Montevideo, Uruguay. METHODS: Eight hundred and one patients were interviewed, including 252 certified cases of BC and 549 frequency-matched controls. Blood groups (ABO, Rh) were obtained from medical records. Multivariate analyses were performed, adjusting for age, selected menstrual and reproductive factors, and family history of BC as well as of other cancers. RESULTS: We found that the absence of Rh factor (Rh-) was positively associated with the risk of BC (adjusted Odds Ratio [OR]=1.49, 95% Confidence Interval [95% CI] 1.05-2.11). Stratified analyses by family history of BC showed a strong association for Rh- with a positive history of first degree relatives (OR=3.17, 95% CI 1.06-9.47). Also stratified analyses by family history of other cancers showed a positive association for Rh- with a positive history of first degree relatives (OR=2.08, 95% CI 1.05-4.11). CONCLUSION: Regarding the implications of an inherited factor like Rh and its associations with the family history of BC, it might increase the probability to generate high-risk individuals if further studies confirm the present preliminary findings. PMID- 19188030 TI - Uteroplacental insufficiency affects kidney VEGF expression in a model of IUGR with compensatory glomerular hypertrophy and hypertension. AB - Low nephron endowment secondary to intrauterine growth restriction (IUGR) results in compensatory hypertrophy of the remaining glomeruli, which in turn is associated with hypertension. However, gender differences exist in the response of the kidney to injury, and IUGR female offspring seems protected from an unfavorable outcome. We previously reported differences in gender-specific gene expression in the IUGR kidney as well as increased circulating corticosterone levels following uteroplacental insufficiency (UPI). Vascular endothelial growth factor (VEGF), which is critical for renal development, is an important candidate in the IUGR kidney since its expression can be regulated by sex-steroids and glucocorticoids. We hypothesize that IUGR leads to altered kidney VEGF expression in a gender-specific manner. Following uterine ligation in the pregnant rat, UPI decreases renal VEGF levels in male and female IUGR animals at birth and through postnatal day 21. However, by day 120 of life, IUGR females have increased kidney VEGF expression, not present in the IUGR males. In addition, IUGR males exhibit increased serum testosterone levels as well as proteinuria. These findings are intriguing in light of the difference in glomerular hypertrophy observed: IUGR males show increased glomerular area when compared to IUGR females. In this model characterized by decreased nephron number and adult onset hypertension, UPI decreases renal VEGF expression during nephrogenesis. Our most intriguing finding is the increased renal VEGF levels in adult IUGR females, associated with a more benign phenotype. We suggest that the mechanisms underlying renal disease in response to IUGR are most likely regulated in a gender specific manner. PMID- 19188031 TI - Achievement of feeding milestones after primary repair of long-gap esophageal atresia. AB - OBJECTIVES: To determine the pattern of feeding milestones following primary repair of long-gap esophageal atresia (EA). METHOD: A questionnaire based upon well established feeding milestones was used. Children after long-gap EA repair, n=40, were compared from after primary repair to healthy children from birth, n=102. RESULTS: The age when surveyed of the EA group and controls was different: 6.2+/-4.7 (mean+/-standard deviation) years, range 1.1-20.9, versus 2.5+/-2.4 years, range 0.0-12.1, p=0.00. The esophageal gap length in the EA group was 5.1+/-1.2 cm and age at repair was 5.5+/-5.0 months. There was no statistically significant difference between the atresia group and controls for feeding milestones; Self feeding finger foods approached significance. There was, however, greater variability in the timing of milestones in the atresia group compared to controls. Feeding milestones were negatively correlated with age at primary repair: drinking with a covered sippy cup, rho=-0.51, p=0.01 and self feeding finger foods, rho=-0.36, p=0.04 were statistically significant. Drinking from a cup correlated with gestational age, rho=0.38, p=0.04, and negatively correlated to esophageal gap length, rho=-0.45, p=0.01. CONCLUSIONS: Despite delayed onset of feeding, major milestones after EA repair occurred in similar pattern to normal infants. An early referral for primary repair is beneficial for earlier acquisition of milestones for infants with long-gap EA. PMID- 19188032 TI - Indices of fetal development derived from heart rate patterns. AB - BACKGROUND: The fetal precursors of mental and cardiovascular disease caused by adverse prenatal environmental influences and manifesting in later age are an important issue of developmental medicine. However the number of measurable functional parameters of a fetus is limited. Evaluation of key parameters involving fetal autonomic control could permit an earlier detection of developmental problems and improved therapeutic strategies. Thus far, however, even the maturation of normal autonomic control has not been sufficiently assessed. AIM: The objective of the present work is to describe normal fetal maturation based on indices of autonomic heart rate modulation. STUDY DESIGN: Heart beat interval series were magnetocardiographically recorded with 1 kHz sampling rate over 30 min in 78 normal fetuses, gestational age (GA) 23-40 weeks. Indices considered were: number of accelerations (AC) and decelerations (DC), RMSSD, SDNN, and short-term/long-term autonomic information flows (AIF_NN, AIF_fVLF). These were measured from the entire 30 min data sets and from activity specific subsets (10 min). RESULTS: In the 30 min recordings: the number of AC increased, number of DC decreased, rMSSD and SDNN increased and AIF_fVLF increased with GA, but AIF_NN remained constant. In the 10 min subsets: SDNN increased in the active state but remained constant in the quiet state and AIF_NN decreased with GA in the quiet state. CONCLUSION: Heart rate patterns from 30 min biomagnetic recordings may provide new indices with which to assess the normal and abnormal maturation of fetal autonomic control and to identify risk of possible disorders in later life. PMID- 19188033 TI - No association between fluctuating asymmetry in highly stabilized traits and second to fourth digit ratio (2D:4D) in human fetuses. AB - Recent studies have suggested that the ratio of the length of the second and fourth digit (2D:4D) may be associated with developmental instability (DI) as measured by the left-right asymmetry of the same digits. Because the 2D:4D ratio is amongst others, determined prenatally as a result of exposure to sex hormones, such an association could indicate that the same prenatal developmental processes determine levels of DI. In this study we criticize these earlier findings and show by simulations that they are confounded by the fact that (non-) linear combinations of the digit lengths are used as both dependent (average asymmetry in digits 2 and 4) and independent (ratio of the lengths of digits 2 and 4) variable. We therefore studied associations between 2D:4D ratios and asymmetry not only in digits but also in several other skeletal elements in deceased human fetuses. In contrast to the earlier studies, we did not find an association between 2D:4D ratios and asymmetry in digits 2 and 4. We argue that this may be due to the low levels of DI in this study, which limits the confounding effects of DI. Also, no associations were detected with the asymmetry of all other trait either. Thus, there appears to be very little evidence of any link between DI and 2D:4D in this population for limb measurements. We conclude that highly stabilized and functionally important traits such as human limbs may in general show limited increases in asymmetry with prenatal stress. PMID- 19188034 TI - Role of nitric oxide on quality of freshly ejaculated bull spermatozoa during heparin-induced in vitro capacitation. AB - The objective of this study was to assess the effects of nitric oxide (NO) on heparin-induced capacitation in vitro of fresh bull sperm, through the addition of Nomega-nitro-l-arginine methyl ester (L-NAME, a NO-synthesis inhibitor) and l arginine (L-Arg, a NO-synthesis precursor) to the capacitation medium. In Experiment 1, different concentrations of L-NAME (0.1, 1, 10mM) and of L-Arg (10mM) were added to the capacitation medium. Sperm motility and vigor were subjectively appraised using direct light microscopy; sperm membrane integrity was examined using a 2% Trypan blue solution while the concentration of nitrate/nitrite (NO(3)(-)/NO(2)(-)) was determined by using the Griess method over a 5h capacitation period. The addition of 10mM L-NAME has inhibited NO synthesis, sperm progressive motility, sperm vigor and sperm membrane integrity (P<0.05) as compared to control. The addition of 10mM L-Arg to the capacitation medium increased all variables evaluated in comparison to the control (P<0.05). In Experiment 2, mitochondrial activity was assessed through the MTT test (3-(4,5 dimetylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), and sperm capacitation was assessed through the test of penetration in homologous oocytes after addition of the 10mM L-NAME, and of the 10mM L-Arg. The addition of 10mM L-NAME caused mitochondrial activity (40%) and the percentage of oocytes penetrated (77%) to decrease in relation to the control (P<0.05). After addition of 0.6mM L-Arg+10mM L-NAME, partial reversal of mitochondrial activity did occur (only 20%). The addition of 10mM L-Arg increased the percentage of oocytes penetrated as compared to control (21%) (P<0.05). These results indicate that: (1) NO is involved in control of progressive sperm motility, vigor, membrane integrity, and mitochondrial activity along the period of heparin-induced capacitation of fresh bovine sperm via NOS/NO; (2) adequate L-Arg/NO concentrations into the capacitation medium can potentiate heparin action or act independently for increasing the number or the quality of capacitated sperm. PMID- 19188035 TI - Application of CdSe nanoparticle suspension for developing latent fingermarks on the sticky side of adhesives. AB - Nanometer-sized fluorescent particles were synthesized in an aqueous solution using TGA (mercaptoacetic acid) as the stabilizer. The prepared solution was characterized by fluorescence spectroscopy and applied for detection of latent fingermarks on adhesives. The effects of stabilizer, precursor, pH value, the concentration of cadmium ion and shell structure on fluorescence were also discussed. The results indicated that the intensity of fluorescence was remarkably increased and the selectivity was well improved when using CdSe nanoparticles, it produced significantly less background development and better contrast after 15 min developing process. PMID- 19188036 TI - Chromatography-tandem mass spectrometry method for the simultaneous quantitation of metoprolol succinate and simvastatin in human plasma. AB - The liquid chromatographic-tandem mass spectrometry method was developed for the accurate quantitation of metoprolol succinate (MET) and simvastatin (SIM) in human plasma which were obtained from the pharmacokinetic (PK) study. The sample purification and pre-concentration was performed by protein precipitation technique using propranolol hydrochloride as working internal standard (WIS). The chromatographic separation was achieved using an isocratic mobile phase consisting of a mixture of acetonitrile and 0.5% formic acid (90:10 (v/v), pH 3.5) flowing through C18 column at a flow rate of 0.2ml/min. Electro spray ionization (ESI) with multiple reaction monitoring (MRM) was used to acquire mass spectra. Ions were monitored in positive mode and the mass transitions measured were m/z 268.1-->m/z 103.2, m/z 441.3-->m/z 325.1 and m/z 260.0-->m/z 129.5 for MET, SIM and WIS, respectively. An extensive pre-study method validation was carried out in accordance with USFDA guidelines. The linearity for the calibration curve in the concentration range of 1.0-500.0 and 0.1-20ng/ml for MET and SIM, respectively and the lower limits of quantitations (LLOQ) were 1.0 and 0.1ng/ml for MET and SIM, respectively. The method was successfully applied to a PK study on fixed dose combination (FDC) tablet containing MET and SIM in healthy human subjects. PMID- 19188037 TI - Identification and structural elucidation of process impurities generated in the end-game synthesis of taranabant (MK-0364) via cyanuric chloride. AB - Taranabant (MK-0364) is a highly potent and selective cannabinoid-1 receptor (CB 1R) inverse agonist. It is being developed at Merck & Company to treat obesity. The chemical synthesis of MK-0364 drug substance involved the direct coupling of chiral amine and pyridine acid side chains mediated by cyanuric chloride. Four major process impurities were observed and characterized using high performance liquid chromatography (HPLC) coupled with ultraviolet (UV) and electrospray ionization (ESI) mass spectrometry (MS) detectors. The exact mass data was used for structural elucidation which suggests that the impurities are derivatives of cyanuric chloride formed in the coupling step. Owing to the reactive nature of these impurities, an interesting degradation phenomenon was observed during stability testing of MK-0364 drug substance when stored at 40 degrees C/75% RH and 25 degrees C/60% RH conditions. Degradation pathways were proposed to explain the changes observed in the HPLC impurity profile. Forced degradation experiments were also conducted to confirm the degradation pathways and assess the stability of the impurities. Finally, the complete stability data of the bulk drug are reported to support the hypothesis. PMID- 19188038 TI - Prediction of the stability of polymeric matrix tablets containing famotidine from the positron annihilation lifetime distributions of their physical mixtures. AB - The aim of the present work was to elucidate the impact of the structural changes of polymeric excipients during the course of storage on the drug release stability of tablets containing different polymers. Matrix tablets were formulated with famotidine as a model drug, using polyvinylpyrrolidone and carbopol matrix. Dissolution tests were carried out before and after storing the tablets under stress conditions for different time intervals. Parameters characterizing the release kinetics of matrix tablets, just as difference and similarity factors, were calculated to compare the release profiles as a function of storage time. Positron annihilation lifetime measurements were carried out to track the structural changes of the physical mixtures containing polymers during the course of storage. The changes in the positron lifetime distribution curves of the famotidine-polymer mixtures were in good correlation with the significant changes of release parameters of tablets. Thus the method would be a valuable tool for the screening of possible destabilizing interactions in the preformulation phase. PMID- 19188039 TI - Lipophilicity of some GABAergic phenols and related compounds determined by HPLC and partition coefficients in different systems. AB - Some phenolic compounds, like propofol and thymol, have been shown to act on the GABA(A) receptor. Taking into account the hydrophobicity of these compounds, their interaction with the membrane surrounding the receptor and consequent non specific effect on receptor modulation cannot be neglected. In the present work, we determined and correlated several lipophilic parameters for both GABAergic agents and three other related phenolic compounds (eugenol, carvacrol and chlorothymol), including logP(o/w), retention data in high performance liquid chromatography (HPLC) by using C18 and immobilized artificial membrane (IAM) columns at different temperatures, and partition coefficients determined in phospholipid liposomes. The correlation results demonstrated the high capacity of the compounds assayed to interact with phospholipid membrane phases, which can be predicted by simple model systems as logP(o/w) or HPLC. The values obtained by HPLC using a fast screening IAM column were the quantitatively closest to the partition coefficients determined in liposome systems, due to the capacity of this column to permit the establishment of molecular interactions like those found in phospholipid membranes. Finally, the fact that all the compounds studied are able to interact with membranes would suggest the participation of some alteration of the GABA(A) receptor lipid environment as part of the receptor modulation exerted by phenolic compounds. PMID- 19188040 TI - The outcomes after surgical resection in pancreatic endocrine tumors: an institutional experience. AB - AIM: Sporadic pancreatic endocrine tumors (PET) can be managed surgically with excellent outcomes. The aim of this study was to analyze surgical outcomes and factors influencing survival. METHODS: Between 1995 and 2007, 96 patients with sporadic PET who underwent surgery at our institution were retrospectively reviewed for clinicopathologic variables and outcomes according to the World Health Organization (WHO) classifications. RESULTS: Thirty-nine patients had well differentiated tumors (WDT) with benign behavior, 23 had uncertain behavior, 27 had low-grade carcinoma, and 7 were diagnosed with high-grade carcinoma. R0 resection was performed in 84 patients. No recurrence was observed in WDT regardless of its behavior or curability but 16 of 34 patients with carcinoma had recurrence. Five-year overall survival (OS) for R0-resected patients with carcinoma was 57%, and OS at 3 years for R1/R2-resected patients was 23% (P = 0.012). The WHO classification and R0 resection were independent prognostic factors in multivariate analysis. CONCLUSIONS: This single institutional experience demonstrated that surgical resection is curative for WDT and recurrences are frequent in spite of curative resection for malignant PET. The WHO classification and R0 resection remained independent prognostic factor. PMID- 19188041 TI - Alternative splinting for flexor tendon rehabilitation with a zone I injury. AB - Many protocols, for zone 1 flexor tendon injuries, encourage early motion of the injured phalanx. Attempts to secure some form of dynamic traction to the distal end of the digit can be the most challenging part of the splint fabrication. It can be even more difficult and painful for the patient when a button is necessary to help secure the tendon. This author has developed what appears to be a relatively pain free, easy method to help resolve this issue. PMID- 19188042 TI - A review of clinical tests and signs for the assessment of ulnar neuropathy. AB - NARRATIVE REVIEW: As part of a comprehensive assessment for suspected ulnar neuropathy, clinical testing plays an important role in the initial identification of a lesion and determining subsequent changes from baseline. The purpose of this article was to review ulnar nerve provocative testing and the substantial collection of diagnostic signs and tests. Administration procedures for each maneuver are described as well as the resulting positive and negative outcomes. The clinical tests described constitute only one aspect of the examination and should not substitute for other key components, such as taking a thorough medical and occupational history. Empirical research studies are indicated to further quantify the relationship between the testing outcomes and the severity of a lesion as well as to determine the most robust motor signs seen in the early stages of the disease. LEVEL OF EVIDENCE: 5. PMID- 19188043 TI - Auditory grip force feedback in the treatment of Writer's cramp. AB - STUDY DESIGN: Pre-post, single-group. INTRODUCTION: Writer's cramp (WC) is a focal dystonia causing impairments in daily life. Behavioral treatment approaches have been shown to improve handwriting performance, though outcomes remain sub optimal. PURPOSE OF THE STUDY: To examine the effects of the handwriting training and auditory grip force feedback in seven patients with WC. METHODS: Handwriting performance was examined before and after treatment. Writing frequency, fluency, and pressure were recorded with a digitizing tablet and grip forces during handwriting were recorded. Subjective writing performance and pain were rated on visual analog scales. RESULTS: The treatment resulted in significant reductions in writing pressure and pain, while writing performance was significantly improved. CONCLUSIONS: Patients in this study with WC, who exhibit grip force and pressure problems, benefit from feedback-supported handwriting training. PMID- 19188044 TI - Induction of anti-whole GAD65 reactivity in vivo results in disease suppression in type 1 diabetes. AB - Most type 1 diabetes mellitus is caused by autoimmune pancreatic beta-cell destruction. Several antigens such as insulin, glutamic acid decarboxylase (GAD) and islet-specific glucose-6-phosphatase catalytic subunit related protein (IGRP) are considered to take part in the autoimmune destructive process. Because the role of GAD in the disease process of type 1 diabetes is still controversial, we investigated the disease phenotype upon in vivo induction of whole GAD65 reactivity using a GAD65 homo knockout NOD splenocytes to NOD-scid transfer system. Splenocytes from 8 to 10-week-old female GAD65 homo knockout (=KOT splenocytes) or age-matched wild type (=WTT splenocytes) NOD mice were transferred into female NOD-scid recipients. As compared to recipients of WTT splenocytes, the onset of diabetes in recipients of KOT splenocytes was significantly delayed (p<0.001). Moreover, TGF-beta expression was enhanced in the pancreas from recipients of KOT splenocytes. Splenocytes from recipients of KOT splenocytes produced IL-10 (/IFN-gamma) upon GAD65 stimulation, whereas those from recipients of WTT splenocytes did not. Based upon these results, we propose that anti-whole GAD65-reactive T cells have the ability to regulate the development of type 1 diabetes. PMID- 19188045 TI - VEGF, survivin and NOS overexpression in psoriatic skin: critical role of nitric oxide synthases. PMID- 19188046 TI - Design and activity of a 'dual-targeted' antimicrobial peptide. AB - Numerous reports have indicated the important role of human normal flora in the prevention of microbial pathogenesis and disease. Evidence suggests that infections at mucosal surfaces result from the outgrowth of subpopulations or clusters within a microbial community and are not linked to one pathogenic organism alone. To preserve the protective normal flora whilst treating the majority of infective bacteria in the community, a tuneable therapeutic is necessary that can discriminate between benign bystanders and multiple pathogenic organisms. Here we describe the proof-of-principle for such a multitargeted antimicrobial: a multiple-headed specifically targeted antimicrobial peptide (MH STAMP). The completed MH-STAMP, M8(KH)-20, displays specific activity against targeted organisms in vitro (Pseudomonas aeruginosa and Streptococcus mutans) and can remove both species from a mixed planktonic culture with little impact against untargeted bacteria. These results demonstrate that a functional, dual targeted molecule can be constructed from a wide-spectrum antimicrobial peptide precursor. PMID- 19188047 TI - MDMA reinstates cocaine-seeking behaviour in mice. AB - BACKGROUND: MDMA effects are mediated by monoaminergic systems, which seem to play a central role in cocaine craving and relapse. METHODS: CD1 mice trained to self-administer cocaine (1 mg/kg/infusion) underwent an extinction procedure in which the cues contingent with drug self-administration remained present. Mice achieving extinction were injected with MDMA (10 mg/kg), d-amphetamine (1 and 2 mg/kg) or saline and tested for reinstatement. RESULTS: Acute MDMA, but not d amphetamine or saline reinstated cocaine-seeking behaviour in mice in which cocaine self-administration and contingent cues were previously extinguished. CONCLUSIONS: Acute MDMA can reinstate cocaine-seeking behaviour in mice. PMID- 19188048 TI - Development of different temoporfin-loaded invasomes-novel nanocarriers of temoporfin: characterization, stability and in vitro skin penetration studies. AB - A previous study revealed that the invasome dispersion containing 3.3% (w/v) ethanol and 1% (w/v) of the terpene mixture (cineole:citral:d-limonene=45:45:10, v/v=standard mixture) could significantly enhance skin penetration of the highly hydrophobic photosensitizer temoporfin (mTHPC). Invasomes enhanced mTHPC deposition in stratum corneum (SC) compared to liposomes without terpenes and conventional liposomes, and they were efficient in delivering mTHPC to deeper skin layers [J. Control. Release 127 (2008) 271-280]. The aim of this study was to develop new mTHPC-loaded invasomes in order to further enhance the drug penetration. The ratio between d-limonene, citral and cineole was varied in the standard terpene mixture and also single terpenes were used. As a result new mTHPC-loaded invasome dispersions were prepared, characterized and investigated for stability and in vitro penetration of mTHPC into abdominal human skin using Franz diffusion cells. Invasomes were of a small particle size (<150nm), high homogeneity (<0.3), mostly unilamellar and spherical, but also deformed vesicles were detected. Invasomes containing 1% (w/v) cineole provided the highest skin penetration enhancement of mTHPC, i.e. they provided high amounts of mTHPC in the SC and deeper skin layers, indicating that also incorporation of a single terpene into invasomes could provide efficient nanocarriers of mTHPC. These invasomes could be considered as a promising tool for delivering the photosensitizer mTHPC to the skin. However, in contrast to most invasomes, being effective nanocarriers of mTHPC, there were also formulations less effective than liposomes containing 3.3% (w/v) ethanol and one formulation was less efficient than conventional liposomes. PMID- 19188049 TI - Fabrication and characterization of ophthalmically compatible hydrogels composed of poly(dimethyl siloxane-urethane)/Pluronic F127. AB - The novel poly(dimethyl siloxane-urethane)/Pluronic hydrogel was fabricated to develop a new ophthalmically compatible material. In this study, the soft segment consists of poly(dimethylsiloxane) dialkanol having hydroxyethyl propoxy end groups and hard segment consists of isophorone diisocyanate (IPDI). The poly(ethylene glycol) methacrylate (PEGMA) was added as the chain-extender to form UV-curable silicone marcomer. Finally, the semi-interpenetrating network (semi-IPN) hydrogel was achieved by reacting with silicone marcomer and Pluronic F127 triblock copolymer under UV-photopolymerization (mSi-PU/F127). It was found that the increase in Pluronic F127 content led to decreased water contact angle and increased the water content of silicone hydrogels. Elastic modulus also decreased with Pluronic F127 content, while surface roughness did not significantly differ from silicone controls. The Pluronic F127 content reached 4%, the apparent protein adsorption amount decreased about 60% in comparison with that of mSi-PU control. It indicated that the mSi-PU/F127 hydrogel membrane had an excellent ability to resist protein adsorption. Additionally, the oxygen permeability (Dk) would decrease 24%, as compared with mSi-PU control. Furthermore, these hydrogel membranes were regarded as non-cytotoxic through in vitro L929 fibroblasts proliferation assay. Overall results demonstrated that the mSi-PU/F127 semi-IPN hydrogel provided silicone hydrogel materials not only having relatively high oxygen permeability and a relatively low modulus, but also enhancing hydrophilicity and anti-protein adsorption. PMID- 19188050 TI - Waist circumference, body mass index and waist to hip ratio for prediction of the metabolic syndrome in Chinese. AB - BACKGROUND AND AIMS: To explore the ability of waist circumference (WC), body mass index (BMI) and waist to hip ratio (WHR) to predict two or more non-adipose components of the metabolic syndrome (MetS) among individuals aged 18-85 in North China. METHODS AND RESULTS: This study is a cluster sample survey of 101,510 individuals, complete data are 75,788 subjects, 59,874 males and 15,914 females. Their ages were 51.9+/-12.7 years (males) and 48.7+/-11.5 years (females). Receiver operating characteristic (ROC) analysis was used to examine discrimination and find optimal cut off values of WC, BMI and WHR to predict two or more non-adipose components of MetS. The area under the ROC curve (AURC) for WC (0.694) and BMI (0.692) in females showed no difference. In males BMI (0.657) had a better discrimination than WC (0.634). WHR was weaker in both sexes. The optimal cut off value of WC in males (86.5 cm) was higher than in females (82.1cm); and that of BMI was about 24 kg/m(2) in both genders. The optimal cut off values of WC, BMI, and WHR, increased with age in both sexes. CONCLUSIONS: BMI and WC are more useful than WHR for predicting two or more non-adipose components of MetS. Cut off values for WC in males, and those of BMI and WHR in both sexes are lower than that in present MetS criteria; WC in females is slightly higher. Cut off values of WC, BMI and WHR were increased with age in the Chinese. PMID- 19188051 TI - Protective effects of Chlorpromazine and Verapamil against cadmium-induced kidney damage in vivo. AB - Overexposure to cadmium (Cd) can induce kidney damage, which was related to the oxidative damage and disturb intracellular Ca2+ homeostasis. Chlorpromazine (CPZ), targeting calmodulin (CaM), and the Ca2+ channel blocker Verapamil (Ver) are involved in intracellular Ca2+ homeostasis processes. The aim of the study was to investigate the kidney damage caused by Cd administrated for 6 weeks and to evaluate the effects of pre-treatment with either chlorpromazine or verapamil on Cd-induced kidney damage. Thirty-two Wistar rats were divided randomly into 4 groups by weight, i.e., control group, Cd-treated group, and CPZ or Ver pre treated group. The Cd-treated group rats were subcutaneously (s.c.) injected with 7micromol CdCl2/kg body weight/day. The CPZ and Ver pre-treated group rats were intraperitoneally (i.p.) injected with 5mg CPZ/kg body weight/day, 4mg Ver/kg body weight/day, respectively, 1h before the s.c. administration of 7micromol CdCl2/kg body weight/day. The control group rats were injected s.c. with saline at the same time. The volume of injection was 2ml/kg body weight, 5 times per week, for up to 6 weeks. After 6 weeks, Cd concentrations in the renal cortex and urine were significantly higher in Cd-treated group than that in controls. Cd concentrations of the urine in CPZ and Ver pre-treated groups were significantly lower than that in Cd-treated group, but there were no significant changes in the renal cortex. Compared with the controls, urinary NAG, ALP activities, and the levels of GSH, MDA, and the activities of PKC, Na(+)-K(+)-ATPase, and Ca(2+) ATPase in rats from the Cd-treated group were significantly increased. SOD activity was suppressed by Cd. Urinary NAG activity and the level of GSH and the activities of PKC and Ca(2+)-ATPase in both CPZ and Ver pre-treated groups were significantly lower than that in Cd-treated rats. The present results showed that Cd-induced kidney damage was related to the oxidative damage and disturb intracellular Ca2+ homeostasis. Both CPZ and Ver possess some ability to prevent cadmium-induced kidney damage via antioxidative action and by maintaining calcium homeostasis. PMID- 19188052 TI - Chronic effects of low lead levels on sperm quality, gonadotropins and testosterone in albino rats. AB - The current study evaluates the impact of low or moderate levels of lead acetate (PbAc) on sperm parameters, gonadotropins (FSH, LH) and testosterone. Adult albino wistar male rats were allocated to five groups and given 0%, 0.025%, 0.05%, 0.1% and 0.3% PbAc in distilled drinking water for 24 weeks. There was no change in body weight gain and in absolute or relative weight of testes, epididymides and seminal vesicles. The ventral prostate weight was decreased in groups exposed to 0.05%, 0.1% and 0.3% PbAc without statistically significant differences. Sperm velocity was decreased in all treated groups while reduction of sperm motility was observed in rats exposed to 0.05%, 0.1% and 0.3% PbAc without statistically significant differences compared to the control group. However, there was a significant increase greater than 100% in the total percentage of abnormal sperm in groups treated with 0.1% (p<0.01) and 0.3% PbAc (p<0.05).The frequency of dead sperm was significantly increased only in the 0.3% PbAc group (p<0.01). Significant increases in frequencies of amorphous head sperm (p<0.01) and abnormal tails (p<0.01) were found in the group exposed to 0.1% PbAc, while the frequency of neck abnormalities was increased in the high-lead exposed group (p<0.01). The levels of LH and FSH were not significantly affected after lead treatment and significant increase in serum testosterone level was noted only in animals administered 0.05% PbAc (p<0.01). In conclusion, our results reveal that some sperm parameters were altered at low or moderate lead concentrations with no obvious alteration of hypothalamic-pituitary function. The increase of testosterone level observed suggests that lead may target testicular function. PMID- 19188053 TI - Adaptogens exert a stress-protective effect by modulation of expression of molecular chaperones. AB - Adaptogens are medicinal plants that augment resistance to stress, and increase concentration, performance and endurance during fatigue. Experiments were carried out with BALB/c mice taking ADAPT-232 forte, a fixed combination of three genuine (native) extracts of Eleutherococcus senticocus, Schisandra chinensis and Rhodiola rosea, characterised for the content of active markers eleutherosides, schisandrins, salidroside, tyrosol and rosavin and in doses of about 30, 90 and 180 mg/kg for seven consecutive days followed by forced swimming test to exhaustion. ADAPT-232 forte strongly augments endurance of mice, increasing the time taken to exhaustion (TTE) in a dose-dependent manner from 3.0+/-0.5 to 21.1+/-1.7 min, approximately seven fold. Serum Hsp72 was measured by EIA both in normal and stressful conditions before and after swimming test. Repeated administration of adaptogen dose dependently increases basal level of Hsp72 in serum of mice from 0.8-1.5 to 5.5-6.3 pg/ml. This effect is even stronger than the effect of stress, including both physical (swimming) and emotional impacts: 3.2+/-1.2 pg/ml. Cumulative effect of stress and adaptogen was clearly observed in groups of animals treated with adaptogen after swimming to exhaustion, when serum Hsp72 increased to 15.1+/-1 pg/ml and remained at almost the same level during the 7 days. It can be concluded that adaptogens induce increase of serum Hsp72, regarded as a defense response to stress, and increase tolerance to stress (in our model combination of physical and emotional stresses). It can be suggested that increased tolerance to stress induced by adaptogen is associated with its stimulation of expression of Hsp70 and particularly with Hsp72 production and release into systemic circulation, which is known as a mediator of stress response involved in reparation of proteins during physical load. Our studies suggest that this could be one of the mechanisms of action of plant adaptogens. PMID- 19188054 TI - Hypoglycemic effect of aspalathin, a rooibos tea component from Aspalathus linearis, in type 2 diabetic model db/db mice. AB - Effects of aspalathin, a green rooibos tea component, on glucose metabolism were studied in vitro and in vivo. We first examined the effect of aspalathin on glucose uptake by cultured L6 myotubes and on insulin secretion from cultured RIN 5F pancreatic beta-cells in vitro, and then investigated the effect of dietary aspalathin on fasting blood glucose level and conducted an intraperitoneal glucose tolerance test (IPGTT) using type 2 diabetes model mice in vivo. Aspalathin dose-dependently and significantly increased glucose uptake by L6 myotubes at concentrations 1-100 microM. It also significantly increased insulin secretion from cultured RIN-5F cells at 100 microM. Dietary aspalathin (0.1-0.2%) suppressed the increase in fasting blood glucose levels of db/db mice for 5 weeks. In IPGTT, aspalathin improved impaired glucose tolerance at 30, 60, 90, and 120 min in db/db mice. These results suggest that aspalathin has beneficial effects on glucose homeostasis in type 2 diabetes through stimulating glucose uptake in muscle tissues and insulin secretion from pancreatic beta-cells. PMID- 19188055 TI - Comparative antiulcer effect of bisdemethoxycurcumin and curcumin in a gastric ulcer model system. AB - The antiulcer effect of bisdemethoxycurcumin, a yellow pigment found mainly in rhizomes of Curcuma longa, was compared with curcumin in gastric ulcer model systems to validate its clinical application as a remedy for peptic ulcer. Western blot analysis of mouse macrophage cell line RAW 264.7 activated with lipopolysaccharide showed that bisdemethoxycurcumin inhibited inducible nitric oxide synthase (iNOS) production significantly but had no effect on tumor necrosis factor-alpha (TNF-alpha) production, whereas curcumin showed stronger suppression of iNOS protein production and inhibited TNF-alpha protein production significantly. However, bisdemethoxycurcumin and curcumin possessed similar potency in scavenging nitric oxide generated from mouse macrophage cell line RAW 264.7. Reverse-transcriptase polymerase chain reaction (RT-PCR) analysis showed that both curcuminoids inhibited the induction of iNOS dose-dependently at the transcriptional level and curcumin also appeared to inhibit the induction of TNF alpha at post-transcriptional level. In an animal model, intraduodenal administration of bisdemethoxycurcumin (5-80 mg/kg body wt.) showed a strong inhibitory effect on gastric acid secretion in pylorus-ligated rats whereas curcumin (5-20 mg/kg body wt.) showed a less inhibitory effect, with maximum potency at a dose of 20mg/kg body wt. Moreover, oral administration of bisdemethoxycurcumin at doses of 20-80 mg/kg body wt. twice daily for 10 days showed a significant curative efficacy in accelerating the healing of acetic acid induced chronic gastric ulcer and promotion of mucosal regeneration in the ulcerated portion in a dose-related manner with potency equal to curcumin. In contrast, the curative potency of curcumin tended to decrease at doses over 160 mg/kg body wt./day. Western blot analysis in ulcerated gastric mucosa showed that bisdemethoxycurcumin dose-dependently reduced the increased protein expression level of iNOS but not TNF-alpha. These results indicated that bisdemethoxycurcumin directly accelerates gastric ulcer healing with potency equal to curcumin. Its antiulcer effect might be due to its properties of decreasing gastric acid secretion and enhancing the mucosal defensive mechanism through suppression of iNOS-mediated inflammation. PMID- 19188056 TI - Regulation of peroxisome dynamics. AB - Peroxisomes are single-membraned organelles ubiquitous to eukaryotic cells that house metabolic reactions that generate and destroy harmful oxidative intermediates. They are dynamic structures whose morphology, abundance, composition, and function depend on the cell type and environment. Perhaps due to the potentially damaging and protective metabolic roles of peroxisomes and their dynamic presence in the cell, peroxisome biogenesis is emerging as a process that involves complex underlying mechanisms of regulated formation and maintenance. There are roughly 30 known peroxins, proteins involved in peroxisome biogenesis, many of which have been conserved from yeast to mammals. This review focuses on the biogenesis of peroxisomes with an emphasis on the regulation of peroxisome formation and the import of peroxisomal matrix proteins in the model organism Saccharomyces cerevisiae. PMID- 19188057 TI - DNA-based amplified electrical bio-barcode assay for one-pot detection of two target DNAs. AB - A sensitive label-free bio-barcode assay provided a PCR-free method for quantitative detection of two nucleic acid targets (HTLV-I and HTLV-II) simultaneously. This DNA biosensor was fabricated with two-component oligonucleotide-modified gold nanoparticles (AuNPs) and two-component oligonucleotide-modified magnetic beads (MBs), which can sandwich a specific target. After liberating the adsorbed thiolated barcode DNA strands (poly A and poly G) from the AuNPs surface with dithiothreitol (DTT) and acidic dipurinization, the electrochemical measurements were directly performed based on the redox activity of guanine (G) and adenine (A) nucleobases. Under the optimal assembling and detection conditions, a good linearity for simultaneous detection was obtained in the range from 4.4x10(-11) to 2.0x10(-9) M, and the detection limit (3sigma) was estimated to be 1.71x10(-12) M for T(1)-DNA and 1.55x10(-12) M for T(2)-DNA. PMID- 19188058 TI - Label-free direct detection of MiRNAs with silicon nanowire biosensors. AB - MicroRNA (miRNA), an 18-24-nucleotide (nt) noncoding RNA molecule in the genes of humans, plants and animals, is emerging as a key player in gene regulation. As a result, label-free, rapid, and sensitive detection for miRNA is of great significance. In this work, a label-free and direct hybridization assay for ultrasensitive detection of miRNA using silicon nanowires (SiNWs) device has been developed. Peptide nucleic acids (PNAs), which serve as a receptor to recognize miRNA directly without labeling the target miRNA, are immobilized on the surface of the SiNW device. Resistance change measured before and after hybridization correlates directly to concentrations of the hybridized target miRNA. Concentration-dependent measurements indicate that a detection limit of 1 fM was obtained using the optimized assay. The technique enables identification of fully matched versus mismatched miRNA sequences. Furthermore, the SiNW device is capable of detecting miRNA in total RNA extracted from Hela cells. This approach paves a way for label-free, early detection of miRNA as a biomarker in cancer diagnostics with very high sensitivity and good specificity. PMID- 19188059 TI - Aptamer-based piezoelectric quartz crystal microbalance biosensor array for the quantification of IgE. AB - The aim of this study was to develop a rapid method to measure IgE in human serum by use of a direct aptamer-based biosensor based on a quartz crystal microbalance (QCM). An avidin monolayer was applied to immobilize aptamers specific for IgE on the gold surface of a quartz crystal. The frequency shifts (FS) of the QCM were measured and related to IgE concentrations. We could demonstrate that aptamers were able to detect IgE with high specificity and sensitivity in 15 min. A linear relationship existed between the FS (Hz) and the IgE concentrations from 2.5 to 200 microg/L in buffer and human serum. The regression equation was y = 1.03x - 0.06 for this QCM method and chemiluminescence in 50 clinical human serum samples. In addition, the aptamer receptors tolerated repeated affine layer regeneration after ligand binding and recycling of the biosensor with little loss of sensitivity. When stored for 3 weeks, the FS were all greater than 90% of those on the response at the first day. The QCM biosensor can measure IgE and offer advantages of high specificity, reusability, low detection limit, no label or sample pretreatment, and low sample requirement. The aptamer QCM biosensor was suitable for sensitive and specific protein detection, representing an innovative tool for future proteomics. PMID- 19188060 TI - Intravenous dexmedetomidine as an adjunct for labor analgesia and cesarean delivery anesthesia in a parturient with a tethered spinal cord. AB - For parturients desiring labor analgesia who have contraindications to neuraxial techniques, intravenous opioid-based patient-controlled analgesia (IVPCA) offers a reasonable alternative, although incomplete analgesia and maternal and neonatal respiratory depression can occur. Dexmedetomidine, a highly selective alpha(2) agonist with negligible placental transfer, may be a valuable adjunct to IVPCA by providing additional analgesia without the respiratory depression associated with increasing opioid usage. The successful use of a dexmedetomidine infusion as an adjunct to unsatisfactory fentanyl IVPCA is reported in a 31-year-old parturient with spina bifida occulta and a tethered spinal cord reaching L5-S1. Dexmedetomidine significantly improved the analgesic quality; increased sedation was observed, but the patient was easily rousable to verbal stimuli. No episodes of maternal hypotension or bradycardia, or fetal heart rate irregularities occurred. Cesarean delivery was required for prolonged first stage of labor and presumed chorioamnionitis; it was conducted under general anesthesia during which the dexmedetomidine infusion was continued. A healthy baby boy was delivered with normal Apgar scores and no discernible neurobehavioral or other deficits. PMID- 19188061 TI - A randomised Phase III trial of glufosfamide compared with best supportive care in metastatic pancreatic adenocarcinoma previously treated with gemcitabine. AB - PURPOSE: There are currently no approved therapies for patients with metastatic pancreatic adenocarcinoma previously treated with gemcitabine. This Phase III trial evaluated the efficacy and safety of glufosfamide as compared with best supportive care (BSC) in this patient population. METHODS: Patients were randomised to glufosfamide plus BSC or to BSC alone with baseline performance status as a stratification factor. The primary end-point was overall survival. RESULTS: Three hundred and three patients were randomised: 148 to glufosfamide plus BSC and 155 to BSC alone. There was an 18% increase in overall survival for glufosfamide that was not statistically significant: hazard ratio (HR) 0.85 (95% confidence interval (CI) 0.66-1.08, p=0.19). Median survival was 105 (range 5 875) days for glufosfamide and 84 (range 2+ to 761) days for BSC. Grade 3/4 creatinine increase occurred in 6 patients on glufosfamide, including 4 with dosing errors. CONCLUSION: These results suggest low activity of glufosfamide in this very refractory patient population. PMID- 19188062 TI - Isolation of bioactive peptides from tryptone that modulate lipase production in Yarrowia lipolytica. AB - In this work the effect of several organic nitrogen sources on lipase production in Yarrowia lipolytica LgX64.81 overproducing mutant was studied. Among them, tryptone and peptone showed the most prominent stimulatory effect. Interestingly, only tryptic and peptic casein digest were found to highly induce lipase biosynthesis while lipase production was very limited in the presence of casein digest from papain and pronase-catalysed hydrolysis and absent in case of chymotryptic digest. It was also demonstrated that the stimulatory peptides should be present in the culture medium at specific proportions and molecular size to match the physiological requirement of Yarrowia lipolytica strain for lipase biosynthesis. Herein, the lipase-production stimulatory peptides were isolated by ion exchange chromatography for the first time. These results had contributed to gain an insight on tryptone role in lipase production by Yarrowia lipolytica. Moreover the use of a chemically defined medium supplemented with the isolated peptides, will improve the efficiency of the process for lipase production in this yeast. PMID- 19188063 TI - Identification of novel and potent 2-amino benzamide derivatives as allosteric glucokinase activators. AB - The identification and structure-activity-relationships (SARs) of novel 2-amino benzamide glucokinase activators are described. Compounds in this series were developed to be potent GK activators, and their binding mode to the GK protein was determined by crystal structure analysis. In vivo pharmacokinetic and acute in vivo efficacy studies of compound 18 are also described. PMID- 19188064 TI - Polyketide synthase thioesterases catalyze rapid hydrolysis of peptidyl thioesters. AB - Polyketide synthase (PKS) thioesterases (TEs) catalyze the macrocyclization of linear acyl chains into macrolactones. Herein we show that peptide based substrates are processed by PKS TEs with greater catalytic efficiency than more native like acyl substrates. This result strengths the link between PKS and non ribosomal peptide synthetase systems and provides a new tool for studying PKS TEs. PMID- 19188065 TI - 3-Acrylamide-4-aryloxyindoles: synthesis, biological evaluation and metabolic stability of potent and selective EP3 receptor antagonists. AB - A series of potent and selective EP(3) receptor antagonists are described. Utilizing a pharmacophore model developed for the EP(3) receptor, a series of 3,4 disubstituted indoles were found to be efficient ligands for this target. These compounds showed high selectivity over IP, FP and other EP receptors. An optimized molecule 7c featured a sound profile and potency in the functional rat and human platelet aggregation assays. PMID- 19188066 TI - (1,2,3-Triazol-4-yl)benzenamines: synthesis and activity against VEGF receptors 1 and 2. AB - Derivatives of (1,2,3-triazol-4-yl)benzenamines are described as potent and ATP competitive inhibitors of vascular endothelial growth factor receptors I and II (VEGFR-1/2). A number of compounds exhibited VEGFR-2 and VEGFR-1 inhibitory activity comparable to that of Vatalanib in both HTRF enzymatic and cellular assays. PMID- 19188067 TI - Walking energy expenditure in able-bodied individuals: a comparison of common measures of energy efficiency. AB - Assessment of walking energy efficiency is potentially useful for objectively quantifying gait efficiency; however, issues regarding methodology, day-to-day variability, inter subject variability (age and size differences) and clinical relevance prevents the assessment of walking energy efficiency from being used as a standard assessment tool. Volume of oxygen (VO(2)) and heart rate were assessed to determine the intra-subject (day-to-day) variability, the impact of age and body parameters and the clinical relevance of gross cost, Net non-dimensional Cost (NNcost) and Energy Efficiency Index (EEI) in able-bodied individuals. Gross cost demonstrated the least amount of day-to-day variability, in comparison to NNcost and EEI. Age and body parameters contributed more to the variability in gross cost than NNcost and EEI. Only net non-dimensional oxygen consumption was related to the velocity of walking. EEI did not correlate with direct measures of oxygen cost. The results from this study demonstrate that utilization of NNcost, a measure that subtracts resting oxygen consumption from walking oxygen consumption and scales for body parameters, is preferable to the traditional measures for comparing individuals of different ages and sizes and is related to the speed that the individual walks. PMID- 19188068 TI - Magnetic resonance imaging of in vivo patellofemoral kinematics after total knee arthroplasty. AB - Simulated partial weight bearing during magnetic resonance imaging of the knee was used to measure patellar tilt, medial-lateral patellar shift, and patellofemoral contact area in three groups of subjects; patients with posterior cruciate retaining (PCR) TKA, patients with bicruciate substituting (BCS) TKA, and healthy controls. Contact stress was also calculated based on the contact area and body weight-based estimates of contact force. Contact stress was significantly (p<0.05) higher in PCR knees (2.5+/-3.0 MPa) than in BCS knees (0.2+/-0.1 MPa) when knees were fully extended, but this difference was not significant (3.7+/-3.5 MPa for PCR knees vs. 1.4+/-1.9 MPa for BCS knees; p>0.05) in early flexion. The results also indicate that patellar tilt (normal=2.4 degrees +/-4.8 degrees, BCS=5.5 degrees +/-5.5 degrees, PCR=-3.0 degrees +/-6.9 degrees change in lateral tilt when moving from full extension to early flexion) and contact area (full extension: normal=267+/-111 mm(2), BCS=344+/-201 mm(2), PCR=83+/-80 mm(2); early flexion: normal=723+/-306 mm(2), BCS=417+/-290 mm(2), PCR=246+/-108 mm(2)) in BCS TKA mimic those in the normal knees more closely than PCR knees do. These results suggest that the patellar component in BCS TKA may be expected to experience less wear than the patellar component in PCR TKA over time. PMID- 19188069 TI - Tibial component overhang following unicompartmental knee replacement--does it matter? AB - As implants are made in incremental sizes and usually do not fit perfectly, surgeons have to decide if it is preferable to over or undersize the components. This is particularly important for unicompartmental knee replacement (UKR) tibial components, as overhang may cause irritation of soft tissues and pain, whereas underhang may cause loosening. One hundred and sixty Oxford UKRs were categorised according to whether they had minor (<3 mm, 70%) or major (>or=3 mm, 9%) tibial overhang, or tibial underhang (21%). One year post surgery, there was no significant difference in outcome between the groups. Five years after surgery, those with major overhang had significantly worse Oxford Knee Scores (OKS) (p=0.001) and pain scores (p=0.001) than the others. The difference in scores was substantial (OKS=10 points). There was no difference between the 'minor overhang' and the 'underhang' group. We conclude that surgeons must avoid tibial component overhang of 3 mm or more, as this severely compromises the outcome. Although this study showed no difference between minor overhang or underhang, we would advise against significant underhang because of the theoretical risk of component subsidence and loosening. PMID- 19188070 TI - Synthesis and in vitro antitubercular activity of a series of quinoline derivatives. AB - A series of 33 quinoline derivatives have been synthesized and evaluated for their in vitro antibacterial activity against Mycobacterium tuberculosis H(37)Rv using the Alamar Blue susceptibility test and the activity expressed as the minimum inhibitory concentration (MIC) in microg/mL. Compounds 5e and 5f exhibited a significant activity at 6.25 and 3.12 microg/mL, respectively, when compared with first line drugs such as ethambutol and could be a good starting point to develop new lead compounds in the fight against multi-drug resistant tuberculosis. PMID- 19188072 TI - Synthesis and pharmacological activities of some sesquiterpene quinones and hydroquinones. AB - Synthesis of protected siphonodictyal C was achieved via drim-7-en-11-al. Some sesquiterpene quinones and hydroquinones were tested for their pharmacological activities in assays in search of antiproliferative, cytotoxic, antiphlogistic, antirheumatic and anti-inflammatory drugs. Wiedendiol B is a ten times stronger cyclooxygenase-2 inhibitor than the reference compound indomethacine. Cyclooxygenase-2 inhibitors are drugs with antiphlogistic and antirheumatic activity. PMID- 19188073 TI - Scanning electron microscopy of Ancylostoma spp. dog infective larvae captured and destroyed by the nematophagous fungus Duddingtonia flagrans. AB - The interaction between the nematode-trapping fungus Duddingtonia flagrans (isolate CG768) against Ancylostoma spp. dog infective larvae (L(3)) was evaluated by means of scanning electron microscopy. Adhesive network trap formation was observed 6h after the beginning of the interaction, and the capture of Ancylostoma spp. L(3) was observed 8h after the inoculation these larvae on the cellulose membranes colonized by the fungus. Scanning electron micrographs were taken at 0, 12, 24, 36 and 48 h, where 0 is the time when Ancylostoma spp. L(3) was first captured by the fungus. Details of the capture structure formed by the fungus were described. Nematophagous Fungus Helper Bacteria (NHB) were found at interactions points between the D. flagrans and Ancylostoma spp. L(3). The cuticle penetration by the differentiated fungal hyphae with the exit of nematode internal contents was observed 36 h after the capture. Ancylostoma spp. L(3) were completely destroyed after 48 h of interaction with the fungus. The scanning electron microscopy technique was efficient on the study of this interaction, showing that the nematode-trapping fungus D. flagrans (isolate CG768) is a potential exterminator of Ancylostoma spp. L(3). PMID- 19188071 TI - 64Cu-AMD3100--a novel imaging agent for targeting chemokine receptor CXCR4. AB - CXCR4 is a chemokine receptor which has been shown to be exploited by various tumors for increased survival, invasion, and homing to target organs. We developed a one step radiosynthesis for labeling the CXCR4-specific antagonist AMD3100 with Cu-64 to produce (64)Cu-AMD3100 with a specific activity of 11.28Ci/ micromol (417GBq/ micromol) at the end of radiosynthesis. Incorporation of Cu(II) ion into AMD3100 did not change its ability to inhibit cellular migration in response to the (only) CXCR4 ligand, SDF-1/CXCL12. (64)Cu-AMD3100 binding affinity to CXCR4 was found to be 62.7 microM. Biodistribution of (64)Cu-AMD3100 showed accumulation in CXCR4-expressing organs and tissues, a renal clearance pathway, and an anomalous specific accumulation in the liver. We conclude that (64)Cu-AMD3100 exhibits promise as a potential PET imaging agent for visualization of CXCR4-positive tumors and metastases and might be used to guide and monitor anti-CXCR4 tumor therapy. PMID- 19188074 TI - Stereological study of the effects of nandrolone decanoate on the mouse liver. AB - BACKGROUND: The beneficial potential of anabolic steroids comes with undesirable side effects. Short and long term side effects have been demonstrated in many organs, and the liver changes are associated with androgenic anabolic steroids use. Despite the widespread physiological, biochemical and pathological investigation of the effects of androgenic anabolic steroids on the liver, the stereological study of the effects of anabolic androgenic steroids on the liver histological structure has received less attention. AIMS: The present study investigates the nandrolone decanoate (ND) effects, on the liver. METHODS: The liver of the control and ND-treated mouse was fixed, processed, stained and studied using modern stereological methods. RESULTS: The analysis of the data revealed that liver weight and volume increased approximately 19% and approximately 35%, in ND-treated group in comparison with the control group respectively. The total volume of the hepatocytes and sinusoids increased approximately 59% and approximately 116% respectively in treated animals. The total volume of the central veins and portal triad veins decreased approximately 42% and approximately 70%, respectively. The total number of hepatocytes nuclei in experimental group increased approximately 20%. The mean total volume of the connective tissue, arteries and bile ductules in the portal tracts and the mean volume of the hepatocytes and their nuclei did not show any significant changes. CONCLUSION: ND can increase the volume of the liver, mainly due to hyperplasia of the hepatocytes. PMID- 19188075 TI - Second harmonic generation from thick leaves using the two-photon laser scanning microscope. AB - A laser-scanning microscope using second harmonic generation (SHG) as a probe is shown to produce high-resolution images of duckweed Lemna minuta leaves. These leaves are multi-cell layer thick. Second harmonic light is generated by a tightly focused short pulse laser beam and is collected by two objectives yielding forward and backward second harmonic digital images. This measurement shows that the signal of the second harmonic imaging in the forward and backward directions depends on the thickness of the chloroplast and that the forward-SH image was brighter than the backward-SH image. The image intensity also depended on the orientation of the chloroplast in relation to the illuminating polarization direction. Their light-induced re-orientation which was affected by the intensity of the illumination could be observed during the experiments. The novelty of this work is to establish new compact technique in which one can use the SH imaging to investigate the true architecture of the sensitive samples, the unknown samples and the samples which is not producing auto-fluorescence. Moreover, investigation of new or unknown samples needs a long time for looking at details of the sample. Thereby the sample will be exposed for long time to the laser radiation that will cause photobleaching and photodamage. Since the SHG does not undergo photobleaching and photodamage this will be the promising technique for investigating the sensitive and new unknown samples. Then one can move to acquire fluorescence images after complete investigation of the true architecture of the sample. The other advantage of SHG is that it has the ability to image highly ordered structural proteins without any exogenous labels. The SHG is an intrinsic and a coherent process. Imaging of intrinsic compounds avoids the complications of slicing and labeling, and samples can be investigated under physiological conditions. PMID- 19188076 TI - Accuracy of combined maxillary and mandibular repositioning and of soft tissue prediction in relation to maxillary antero-superior repositioning combined with mandibular set back A computerized cephalometric evaluation of the immediate postsurgical outcome using the TIOPS planning system. AB - AIM: The purpose of the present study was to evaluate the immediate postsurgical outcome of planned and predicted hard and soft tissue positional changes in relation to maxillary antero-superior repositioning combined with mandibular set back using the computerized, cephalometric, orthognathic surgical planning system (TIOPS). MATERIAL AND METHODS: Out of 100 prospectively and consecutively treated patients, 52 patients manifested dentofacial deformities requiring bimaxillary orthognathic surgery with maxillary antero-superior repositioning combined with mandibular set back and so were included. All patients were managed with rigid internal fixation (RIF) and without intermaxillary fixation (IMF). Preoperative cephalograms were analyzed and treatment plans and prediction tracings produced by computerized surgical interactive simulation. The planned horizontal and vertical hard tissue positional changes were transferred to model surgery on a three-dimensional articulator system (SAM) and finally to surgery. Five to six weeks after surgery, the actually obtained hard and soft tissue profile changes were cephalometricly assessed. RESULTS: The mean accuracy of the planned and predicted hard and soft tissue outcome was relatively high varying from 0.0mm to 0.5mm from one cephalometric reference point to another. At the cephalometric reference points where significant differences between planned/predicted, and actually obtained hard and soft tissue positional changes were demonstrated, these significant inaccuracies were, except for the predicted horizontal position of the lower lip, relatively small, varying from 0.2mm to 1.1mm. However, the variability of the predicted hard and soft tissue individual outcome was relatively high. CONCLUSION: The current study demonstrates from a mean point of view relatively high predictability of the immediate postsurgical hard and soft tissue outcome. However, as the variability of the predicted individual outcome seems to be relatively high, caution should be taken when presenting the planned and predicted hard and soft tissue positional changes to the individual patient, preoperatively. PMID- 19188077 TI - Neuropsychological function in children with cyanotic heart disease undergoing corrective cardiac surgery: effect of two different rewarming strategies. AB - OBJECTIVE: Hypothermia conventionally used in cardiopulmonary bypass necessitates rewarming to normothermic temperatures, which has been shown to be associated with neuropsychological injury. We studied the effects of two different rewarming strategies on postoperative neuropsychological function in cyanotic paediatric patients undergoing elective primary intracardiac repair of tetralogy of Fallot with the aid of cardiopulmonary bypass. METHODS: This was a randomised clinical study undertaken in the cardiothoracic centre of a tertiary level referral and teaching hospital. Eighty children, aged 6-15 years undergoing elective primary intracardiac repair of tetralogy of Fallot using cardiopulmonary bypass under moderate hypothermia at 28 degrees C were included in this study. The patients were randomly allocated into two groups of 40 each. Group 1 patients were rewarmed to a nasopharyngeal temperature of 33 degrees C while group 2 patients were rewarmed to a nasopharyngeal temperature of 37 degrees C before weaning them off cardiopulmonary bypass. The anaesthetic and bypass management was standardised for both the groups. All patients were assessed for neuropsychological function preoperatively and on the fifth postoperative day using the MISIC tests. The amount of blood loss and need for blood and blood product transfusion postoperatively, need for pacing, increased inotropes or vasodilator use and time to extubation were also noted. Serum s-100beta levels were measured post anaesthetic induction and at 24h postoperatively. RESULTS: There was a significant deterioration in neuropsychological function postoperatively in the patients in group 2 (37 degrees C) as compared to their preoperative function. This was associated with higher s-100beta levels 24h postoperatively in group 2 (37 degrees C) compared to group 1 (33 degrees C) patients. The time to extubation was longer in group 1 (33 degrees C). No significant differences were noted in the amount of postoperative blood loss, blood and blood product use, inotrope or vasodilator use and the need for pacing. CONCLUSION: Weaning off bypass at 33 degrees C is associated with lesser postoperative neuropsychological dysfunction compared to rewarming to 37 degrees C before weaning off bypass. This may be used as a tool to decrease neurologic morbidity following cardiac surgery in children with congenital cyanotic heart disease. PMID- 19188079 TI - Chest wall resection for lung cancer: indications and techniques. AB - Lung cancer invasion of the chest wall is not a common challenge and represents only about 5% of all patients resected for lung cancer. In T3N0M0 tumours, long term survival reaches 40-50%, provided certain conditions are fulfilled. The number of explorative thoracotomies and the rate of non-radical resections might be high due to the local extension of an aggressive tumour. Mortality after resection is as high as for pneumonectomy. For historical and anatomical reasons, we have to divide the patients into two groups: infiltration of, and above, the second rib (Pancoast) and tumours located caudally to the second rib. We have to define the two entities. There is a problem concerning correct diagnosis: many tumours reach the chest wall. If the lung is not adherent to the parietal pleura, a standard lobectomy can be performed. However, in the case of adhesions, the differentiation between tumour invasion and inflammation may be difficult. We do not want to perform over-treatment since lung resection en bloc with the chest wall has a higher morbidity and mortality than lobectomy. But we have to avoid opening the tumour intraoperatively or perform a non-radical resection. Therefore, we need a preoperative diagnostic tool answering the question of extrapulmonary infiltration. In this context, we will discuss whether extrapleural lung resection is acceptable in the case of pleural invasion without chest wall involvement. The prognosis of patients with tumours invading the chest wall and mediastinal lymph node metastasis is worse. But patients with ipsilateral supraclavicular lymph node metastasis are not excluded. Thus, careful clinical investigations are necessary. To achieve complete resection, the surgeon should use anatomical knowledge to choose the best form of access to make radical resection more feasible. The problem of pain after thoracotomy is accentuated after chest wall resection, especially after paravertebral resection. The use of modern pain treatment is very important. PMID- 19188080 TI - Pain therapy. AB - Cancer-related pain is a major issue of healthcare systems worldwide. The reported incidence, considering all stages of the disease, is 51%, which can increase to 74% in the advanced and terminal stages. For advanced cancer, pain is moderate to severe in about 40-50% and very severe or excruciating in 25-30% of cases. Pain is both a sensation and an emotional experience. Pain is always subjective; and may be affected by emotional, social and spiritual components thus it has been defined as "total pain". From a pathophysiological point of view, pain can be classified as nociceptive (somatic and visceral), neuropathic (central, peripheral, sympathetic) idiopathic or psychogenic. A proper pain assessment is fundamental for an effective and individualised treatment. In 1986 the World Health Organisation (WHO) published analgesic guidelines for the treatment of cancer pain based on a three-step ladder and practical recommendations. These guidelines serve as an algorithm for a sequential pharmacological approach to treatment according to the intensity of pain as reported by the patient. The WHO analgesic ladder remains the clinical model for pain therapy. Its clinical application should be employed only after a complete and comprehensive assessment and evaluation based on the needs of each patient. When applying the WHO guidelines, up to 90% of patients can find relief regardless of the settings of care, social and/or cultural environment. This is the standard treatment on a type C basis. Only when such an approach is ineffective are interventions such as spinal administration of opioid analgesics or neuroinvasive procedures recommended. PMID- 19188082 TI - Ultrafast shape recognition: evaluating a new ligand-based virtual screening technology. AB - Large scale database searching to identify molecules that share a common biological activity for a target of interest is widely used in drug discovery. Such an endeavour requires the availability of a method encoding molecular properties that are indicative of biological activity and at least one active molecule to be used as a template. Molecular shape has been shown to be an important indicator of biological activity; however, currently used methods are relatively slow, so faster and more reliable methods are highly desirable. Recently, a new non-superposition based method for molecular shape comparison, called Ultrafast Shape Recognition (USR), has been devised with computational performance at least three orders of magnitude faster than previously existing methods. In this study, we investigate the performance of USR in retrieving biologically active compounds through retrospective Virtual Screening experiments. Results show that USR performs better on average than a commercially available shape similarity method, while screening conformers at a rate that is more than 2500 times faster. This outstanding computational performance is particularly useful for searching much larger portions of chemical space than previously possible, which makes USR a very valuable new tool in the search for new lead molecules for drug discovery programs. PMID- 19188083 TI - Screening for X-linked creatine transporter (SLC6A8) deficiency via simultaneous determination of urinary creatine to creatinine ratio by tandem mass spectrometry. AB - High urinary creatine to creatinine ratio (U-CrCrtR) is a potential diagnostic marker of X-linked creatine transporter (SLC6A8) deficiency. We developed a tandem mass-spectrometry method to simultaneously determine urinary creatine and creatinine in 975 individuals (0-18 years). U-CrCrtR increased up to 8 years and decreased thereafter. U-CrCrtR was 2.29 and 2.12 (99th percentile: 1.87) in two males with subsequently confirmed SLC6A8 mutations. The frequency of SLC6A8 deficiency was 2.3% in 157 males at risk. PMID- 19188081 TI - Y-Family DNA polymerases may use two different dNTP shapes for insertion: a hypothesis and its implications. AB - Chemicals and radiation can damage DNA leading to the formation of adducts/lesions, which - if not removed by DNA repair pathways - usually block replicative DNA polymerases (DNAPs). To overcome such potentially lethal blockage, cells have lesion bypass DNAPs, which are often in the Y-Family and include several classes. One class includes human DNAP kappa and E. coli DNAP IV, and they insert dCTP in the non-mutagenic pathway opposite [+ta]-B[a]P-N(2)-dG, which is the major adduct formed by the environmental carcinogen benzo[a]pyrene. Another class includes hDNAP eta and ecDNAP V, and they insert dATP opposite [+ta]-B[a]P-N(2)-dG in the dominant G-->T mutagenic pathway. Herein we develop a hypothesis for why the IV/kappa-class preferentially does cellular dCTP insertion. On the minor groove side of the active site, Y-Family DNAPs have a cleft/hole that can be analyzed based on an analogy to a "chimney." Our models of DNAP IV show a large chimney opening from which the pyrene of [+ta]-B[a]P-N(2)-dG can protrude, which allows canonical adduct-dG:dCTP pairing. In contrast, our models of DNAP V have small chimney openings that forces adduct-dG downward in the active site such that canonical adduct-dG:dCTP pairing is not possible. Based on X-ray structures, sequence alignment and our modeled structures of Y-Family DNAPs, chimney opening size seems primarily controlled by one amino acid ("flue handle"), which dictates whether nearby amino acids ("flue") plug the chimney or not. Based on this analysis, a correlation is apparent: the flue is closed in V/eta-class DNAPs giving small chimney openings, while the flue is open for the IV/kappa-class giving large chimney openings. Secondarily, a hypothesis is developed for why the V/eta-class might preferentially do cellular dATP insertion opposite [+ta]-B[a]P-N(2)-dG: the small chimney forces adduct-dG lower in the active site, possibly leading to catalysis using a non-canonical dNTP shape that permits syn-adenine:adduct-dG base pairing. In summary, a hypothesize is developed that the pyrene moiety of [+ta]-B[a]P-N(2)-dG protrudes from the large chimney opening of DNAP IV, thus permitting canonical dCTP:adduct-dG pairing, while the small chimney opening of DNAP V forces [+ta]-B[a]P-N(2)-dG lower down in the active site, in which syn-adenine can pair with adduct-dG via a non canonical dNTP shape. PMID- 19188084 TI - Mycobacterium tuberculosis and Mycobacterium africanum in stools from children attending an immunization clinic in Ibadan, Nigeria. AB - BACKGROUND: Tuberculosis is a major cause of childhood morbidity and mortality in Nigeria. Diagnosis of childhood tuberculosis is a global challenge making early treatment a mirage. In this study we investigated the stools of children for the presence of mycobacteria. METHODS: Stool samples from children aged 3 days to 3 years who presented for postnatal immunization at a large university-based clinic in Nigeria, were subjected to Ziehl-Neelsen staining. Samples with acid-fast bacilli were further processed using mycobacterial culture, spoligotyping, and deletion typing. RESULTS: One hundred and ninety-two stool samples from different children were collected and processed. Thirty (15.6%) had acid-fast bacilli. Of these, eight had Mycobacterium tuberculosis and one had Mycobacterium africanum. CONCLUSIONS: Approximately 5% (9/192) of apparently well children had evidence of potentially serious tuberculosis infection. The usefulness of stool specimens for diagnosing pediatric tuberculosis warrants further investigation. PMID- 19188085 TI - Biering-Sorensen test scores in coal miners. AB - OBJECTIVES: Biering-Sorensen test is an isometric back endurance test. Biering Sorensen test scores have varied in different cultural and occupational groups. The aims of this study were to collect normative data on Biering-Sorensen holding times, to determine the discriminative ability of the Biering-Sorensen test in Turkish coal miners, and to examine the association between Biering-Sorensen test result and functional disability. METHODS: One hundred and fifty male coal miners participated in this study. Trunk extensor muscle strength was measured using the Biering-Sorensen test. Oswestry disability index was used to measure the functional disability level of low back pain. RESULTS: The mean Biering-Sorensen holding time for the total subject group was 107.3+/-22.5s. The mean time of Biering-Sorensen test of the subjects with and without low back pain were 99.9+/ 19.8 and 128.6+/-15.2 s, respectively. The difference between the subjects with and without low back pain was statistically significant (p<0.001). There was a statistically significant negative correlation between Oswestry functional disability score and Biering-Sorensen holding time (r=-0.824, p<0.001). CONCLUSIONS: Turkish coal miners have low mean back extensor endurance holding times. Biering-Sorensen test had a good discriminative ability in our study group. Trunk muscle strength has a significant effect on the disability level of low back pain. Thus trunk muscle endurance training exercise therapy may be effective for the reduction of disability in patients with low back pain. PMID- 19188086 TI - Cortical and interfacial bone changes around a non-cemented hip implant: simulations using a combined strain/damage remodelling algorithm. AB - Prostheses alter the loading pattern in bones. For example femoral hip implants cause the cortical bone to remodel and the implant/bone interface to react. To date, simulations of peri-prosthetic bone adaptations have succeeded for the bulk bone remodelling only. In this study we use an approach based on a combined strain/damage algorithm to simultaneously predict both bulk and interfacial peri prosthetic bone adaptation around a non-cemented hip prosthesis. The influence of stem stiffness is investigated; a flexible iso-elastic stem (20GPa), an titanium alloy stem (110GPa), and a stiff cobalt chrome (CoCrMo) stem (210GPa). The results predict that an iso-elastic stem reduces proximal bone loss because stress shielding is prevented but it increases proximal interface resorption due to damage-stimulated resorption. On the other hand, a stiff cobalt chrome stem increases proximal strain-stimulated resorption but does not induce proximal interfacial damage-stimulated resorption; however damage-stimulated resorption surrounding the distal tip is predicted to increase with stiffer stems. Simulations for the titanium stem were predicted to minimise both strain and damage related remodelling. We propose that this combined strain/damage remodelling algorithm can provide realistic simulations of the response of bone around load-bearing orthopaedic implants. PMID- 19188087 TI - Behaviour modification in the management of chronic habits of abnormal eye rubbing. AB - OBJECTIVES: To describe how and why many keratoconus patients do not comply with strong advice to control chronic habits of abnormal rubbing. To outline a behaviour modification approach for controlling chronic habits of abnormal rubbing. METHODS: Common reasons for chronic habits of abnormal rubbing have been reviewed as a basis for specifying a behavioural modification approach to habit reversal. RESULTS: The methods described are organized into the classic behavioural modification structure of: (1) habit awareness, (2) competing responses, (3) development of motivation, and (4) social support. This structure is supported by the application of social influence principles to achieve optimum compliance. CONCLUSIONS: The use of take-home written information in the form of an Abnormal Rubbing Guide is the basis for the development of motivation. Family social support is based upon a widening of the responsibility for avoiding eye rubbing to all family members. Some patients will need minimal application of these principles with patient education being sufficient intervention to achieve habit reversal. For patients with strong provocation to rubbing and/or by having a well established rubbing habit, a greater exposure to the habit reversal program described is indicated. Successful habit reversal may slow the rate of ectasia progression. Prophylactic application of the methods described for patients who are at risk for developing keratoconus, or post-laser assisted in situ keratomileusis keratectasia, may show that some forms of keratectasia are preventable. PMID- 19188088 TI - Tunability of anion binding strength based on acyl-thiourea receptors containing isatin group. AB - Some acyl-thiourea derivatives containing isatin group were synthesized and their interactions with anions were investigated using UV-vis spectroscopy and (1)H NMR titrations in DMSO and DMSO-d(6), respectively. These compounds have a same molecular framework, functionalising with different groups lead to different anion binding strength of these receptors. Receptor 1 showed a higher binding affinity for AcO(-) than for F(-), due to the cooperative multiple hydrogen bond interactions of AcO(-) with the acyl-thiourea group and N-H group in the indole unit of receptor 1. Displacing the N-H proton in the indole unit with -CH(3) group, receptor 2 showed no obviously discriminative responses for F(-), AcO(-) and H(2)PO(4)(-) due to lack of such additional binding. In the case of receptor 3, which was functionalised with strong electron-withdrawing group, it showed selectively chromogenic response for F(-) based on double deprotonation of the receptor in DMSO, whereas AcO(-) and H(2)PO(4)(-) induced single deprotonation only. PMID- 19188089 TI - Determination of ultra trace amounts of protein by 4-chlorosulfo-(2' hyaroxylphenylazo)-rhodanine-Ti(IV) complex [ClSARP-Ti(IV)] as the fluorescence spectral probe in AOT microemulsion. AB - Experiments indicated that protein can enhance the fluorescence of the 4 chlorosulfo-(2'-hydroxylophenylazo)-rhodanine-Ti(IV) complex [ClSARP-Ti(IV)] in the presence of bis(2-ethylhexyl)sulfosuccinate sodium salt (AOT) microemulsion. Based on this, a sensitive and reproducible fluorometric method for the determination of micro amount protein was developed. The calibration curves of four proteins were given. Under the optimum experimental conditions, the enhanced fluorescence intensity of the system was in proportional to the concentration of protein in the range of 0.1-11 microg mL(-1) for bovine serum albumin (BSA), 1.0 10 microg mL(-1) for human serum albumin (HSA), 1.0-50 microg mL(-1) for ovalbumin (Ova) and 2.5-18 microg mL(-1) for gamma-globulins (gamma-G). Their detection limits were 0.070, 0.071, 0.33 and 0.22 microg mL(-1), respectively. The ClSARP-Ti(IV) complex as a spectral probe can be used to the determination of protein in milk powder and oatmeal yielding with satisfactory results. Therefore, the proposed method is one of the most sensitive methods available. In addition, the interaction mechanism of this system is studied by multi-techniques. PMID- 19188091 TI - The use of teaching portfolios to promote excellence and scholarship in nurse education. AB - Achieving excellence in nurse education and demonstrating scholarship in teaching is a challenge for nurse educators who find themselves torn between maintaining high standards in nurse education and their own need for recognition and promotion in an environment that primarily favours research and publications over excellence in teaching. The use of a teaching portfolio is a way to display excellence in nurse education and to provide a public display of teaching scholarship. While not a new concept in nursing education, it has some originality in its application to practising nurse educators. However, the requirement for and guidelines on the development of a portfolio for nurse educators (other than those undertaking educational training) has been little explored. The aim of this paper is to explore the concept of a teaching portfolio as a vehicle to demonstrate excellence and scholarship among seasoned nurse educators. The process of developing a teaching portfolio at one university setting is outlined, so that other teachers are enlightened to the potential benefits of this activity. PMID- 19188090 TI - Molecular evidence of human T-cell lymphotropic virus types 1 and 2 (HTLV-1 and HTLV-2) infections in HTLV seroindeterminate individuals from Sao Paulo, Brazil. AB - BACKGROUND: Using enzyme immunoassays and Western blot (Wb) tests, HTLV serodiagnosis yields indeterminate results in a significant number of cases. OBJECTIVE: To determine the prevalence of HTLV infection among HTLV seroindeterminate individuals. STUDY DESIGN: We studied peripheral blood mononuclear cells from 65 anti-HTLV Wb-seroindeterminate individuals by attempting to amplify proviral DNA sequences (tax and pol) to identify HTLV-1 and HTLV-2 infections. RESULTS: These 65 specimens exhibited predominantly (43%) anti HTLV antibodies to gag-coded antigens in the absence of anti-p24 on Wb analysis. Tax proviral sequences were detected in 6 (9.2%) samples. According to restricted fragment polymorphism analysis (RFLP), we identified HTLV-1 proviral DNA in 4 samples, HTLV-2 in one and sequences from both in another. Nested PCR for the pol region was positive in 3 (4.6%) specimens, which were also positive for tax sequences. After hybridization HTLV-1 infection was confirmed in 2 samples (3.1%) and HTLV-2 in another (1.5%). Detection of a single HTLV DNA sequence may be due to infection by defective provirus, but its significance remains undefined. In this cohort, no Wb reactivity pattern was predictive of proviral detection. HTLV 1 infection was demonstrated in an individual who had Wb reactivity to gag-coded antigens only. CONCLUSIONS: This emphasizes the importance of clinical and laboratory follow-up of HTLV-seroindeterminate individuals from endemic areas. PMID- 19188092 TI - B-cell activating factor (BAFF) plays a role in the mechanism of action of a tolerogenic peptide that ameliorates lupus. AB - Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by dysregulated immune responses mediated by T and B cells. A tolerogenic peptide, designated hCDR1, ameliorated the serological and clinical manifestations of SLE in mouse models of lupus. We investigated the role of B-cell activating factor (BAFF) in the beneficial effects of hCDR1. BAFF production was reduced in hCDR1 treated mice in association with diminished production of dsDNA-specific autoantibodies and proteinuria levels. In addition, IFN-gamma and IL-10, which induce BAFF secretion, were down-regulated in hCDR1-treated mice. The reduced levels of BAFF correlated with a lower rate of maturation and differentiation of B cells, and with a decrease in integrin expression and anti-apoptotic gene expression by B cells. Moreover, BAFF signaling through the NF-kB pathways was inhibited in hCDR1-treated mice. Thus, down-regulation of BAFF plays a role in the mechanism of action by which hCDR1 ameliorates lupus manifestations. PMID- 19188093 TI - Comparisons of affinities, avidities, and complement activation of adalimumab, infliximab, and etanercept in binding to soluble and membrane tumor necrosis factor. AB - The TNF antagonists adalimumab, infliximab, and etanercept are effective treatments for rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, and psoriasis, but only adalimumab and infliximab have been found to be efficacious in Crohn's disease. The present studies evaluated the TNF-binding and complement-activating properties of adalimumab, infliximab, and etanercept to determine whether these properties may explain differences in their clinical efficacy profiles. Association and dissociation rates of binding to soluble TNF were measured by surface plasmon resonance, and were found to be similar for adalimumab, infliximab, and etanercept, as were their calculated binding affinities. Avidity of binding to soluble TNF, measured by KinExA technology, was 10- to 20-fold greater for soluble etanercept (K(D)=0.4 picomolars [pM]) than for soluble adalimumab or infliximab (K(D)=8.6 and 4.2 pM, respectively). (125)I adalimumab, -infliximab, and -etanercept bound to membrane TNF (mTNF) on mTNF transfected cells with similar affinities (K(D)=483, 468, and 445 pM, respectively) that were each lower than for soluble TNF. Complement-dependent cytotoxicity (CDC) was induced in mTNF-transfected cells by adalimumab and infliximab, but was not induced in activated normal human PBMC by any of the 3 agents. In conclusion, the binding properties of adalimumab, infliximab, and etanercept were similar for soluble TNF, and very similar for mTNF, yet none of the 3 was able to induce CDC in activated PBMC. These results suggest that the different clinical efficacy profiles of these agents are not explained by differences in either TNF-intrinsic binding properties or complement lysis. PMID- 19188094 TI - Mechanisms of adaptive supersensitivity in vas deferens. AB - Adaptive supersensitivity is a phenomenon characteristic of excitable tissues and discloses as a compensatory adjustment of tissue's response to unrelated stimulatory endogenous and exogenous substances after chronic interruption of excitatory neurotransmission. The mechanisms underlying such higher postjunctional sensitivity have been postulated for a variety of cell types. In smooth muscles, especially the vas deferens with its rich sympathetic innervation, the mechanisms responsible for supersensitivity are partly understood and appear to be different from one species to another. The present review provides a general understanding of adaptive supersensitivity and emphasizes early and recent information about the putative mechanisms involved in this phenomenon in rodent vas deferens. PMID- 19188095 TI - Discharge rate profiles of paratrigeminal nucleus neurons throughout a pressor event in non-anaesthetized rats. AB - Located in the lower brainstem, the paratrigeminal nucleus (Pa5) is related to cardiorespiratory autonomic reflex functions. To characterize the structures' role in blood pressure regulation and the cardiovascular reflex responses Pa5 unit activity was evaluated during a phenylephrine-produced pressor response in non-anaesthetized rats by means of simultaneous many-unit recording. Ninety five percent of the identified Pa5 responded to baroreceptor stimulation, 77% increasing and 23% decreasing firing rates. Cross-correlation analysis of neuron electrical behavior referenced to the heart beat event revealed that 65% of the featured cardiac cycle-locked rhythmic activity. The identification of neurons that change firing rates in response to increases of arterial pressure with cardiac cycle-locked rhythmic activity, further supports for a role for the nucleus in moment to moment control of blood pressure. The largest changes in firing rate occurred in the units with low resting firing rates in response to the ascending phase of the pressor event. Thus, the group displaying both cardiac cycle-locked and other rhythmic activities within the ranges of cardiac and respiratory rates or arterial pressure low frequencies, is probably the most influential regarding homoeostatic reflex responses. The findings advance the notion that the dynamic control of blood pressure involves lower brainstem integration of cardiac and respiratory reflexes. PMID- 19188096 TI - Direct electron transfer and electrocatalysis of myoglobin loaded in layer-by layer films assembled with nonionic poly(ethylene glycol) and ZrO(2) nanoparticles. AB - Nonionic poly(ethylene glycol) (PEG) and ZrO(2) nanoparticles were successfully assembled into {PEG/ZrO(2)}(n) layer-by-layer films on solid surfaces through coordination interaction between the ether oxygen groups in PEG and the Zr(IV) in ZrO(2) nanoparticles. The {PEG/ZrO(2)}(n) films were then immersed in myoglobin (Mb) solution at pH 5.0 to load Mb into the films, designated as {PEG/ZrO(2)}(n) Mb. Cyclic voltammetry (CV), quartz crystal microbalance (QCM), and scanning electron microscopy (SEM) were used to characterize both {PEG/ZrO(2)}(n) and {PEG/ZrO(2)}(n)-Mb films. Mb in the {PEG/ZrO(2)}(n)-Mb films fabricated on pyrolytic graphite (PG) electrodes showed direct and quasi-reversible CV response, which could be used to electrocatalyze reduction of oxygen and hydrogen peroxide. The interaction between Mb and {PEG/ZrO(2)}(n) films in loading was also discussed and explored. The results suggest that the electrostatic interaction is the main driving force for the loading of Mb into the {PEG/ZrO(2)}(n) films, while hydrogen bonding and/or hydrophobic interaction are also important factors for stabilizing {PEG/ZrO(2)}(n)-Mb films in blank buffers. The comparative experiments demonstrated that only those heme proteins whose dimension was smaller than the average pore size of the films were able to be loaded into the films and exhibited electroactivity. PMID- 19188097 TI - Determination of visnagin in rat plasma by liquid chromatography with tandem mass spectrometry and its application to in vivo pharmacokinetic studies. AB - A rapid and sensitive method for the determination of visnagin in rat plasma was developed using liquid chromatography tandem mass spectrometry (LC-MS/MS). The plasma samples were processed by protein precipitation with methanol:acetonitrile (25:75) and warfarin was used as the internal standard. After vortex mixing and centrifugation, the supernatants were diluted with water (1:5) and then directly injected onto a Phenomenex Synergi Max RP column (75 mm x 2.0 mm ID, 4 microm) with isocratic elution at a flow rate of 0.2 mL/min. The mobile phase consisted of water and methanol (15:85, v/v) containing 0.1% formic acid and 5mM ammonium acetate. The total run time (injection to injection) was less than 4 min. Detection of the analytes was achieved using positive ion electrospray in the selected reaction monitoring (SRM) mode. The linear standard curve ranged from 1.0 to 5000 ng/mL and the precision and accuracy (inter- and intra-run) were within 4.5% and 4.3%, respectively. The method, which is rapid, simple, and precise, was successfully used to support a visnagin pharmacokinetics study. PMID- 19188098 TI - Measurement of pentosidine in human plasma protein by a single-column high performance liquid chromatography method with fluorescence detection. AB - A rapid and sensitive single-column high-performance liquid chromatography method and application for the detection of protein bound pentosidine is described. Pentosidine, a cross-link between arginine and lysine, is a well-characterized advanced glycation endproduct. In order to detect protein-bound pentosidine, plasma proteins were hydrolysed in 6N HCl. Detection of pentosidine is done based on its own fluorescence characteristics using fluorimetric detection (E(x)=325 nm, E(m)=385 nm). Separation is done, with a run-to-run time of 30 min, on a C(18) Allspehere ODS-II column with a citric acid acetonitrile buffer. This detection enables sensitive and specific determination of protein bound pentosidine in plasma with a detection limit of 2.2 nmol/l or 0.02 pmol/mg protein (signal-to-noise: 6). The intra-assay coefficient variation is 6.5% at a plasma pentosidine concentration of 0.47 pmol/mg protein and 2.0% at a concentration of 1.27 pmol/mg protein. The inter-assay coefficient variation is 3.1% at a plasma pentosidine concentration of 0.43 pmol/mg protein and 1.6% at a concentration of 1.40 pmol/mg protein. Linearity is tested in 4 different plasma samples and showed linearity (0-200 nmol/l, r(2)>0.99). Recovery of pentosidine in 4 different plasma samples at different concentration levels is 102+/-10% (mean+/-SD). Using this method protein bound pentosidine concentration is investigated in healthy controls (n=24, age 67+/-9 years) and patients with end stage renal disease (n=24, age 65+/-10 years). Higher plasma concentrations of protein bound pentosidine are measured in the patient group as compared with the control group 3.05 (2.03-3.92)pmol/mg protein and 0.21 (0.19-0.33)pmol/mg protein, respectively (median (interquartile range), p<0.00001). These results are consistent with previously reported results. PMID- 19188099 TI - Extraction of PCR-ready DNA from Staphylococcus aureus bacteriophages using carboxyl functionalized magnetic nonporous microspheres. AB - Magnetic microspheres P(HEMA-co-EDMA) were used for PCR-ready phage DNA isolation from lysogenic strains of Staphylococcus aureus, including two new clinical isolates. The conditions of phage particle lysis were optimized. The quality of eluted phage DNA was evaluated by PCR. It was demonstrated that PCR-ready phage DNA can be isolated from small volumes of phage lysates (150 microl) by magnetic microspheres. The reported method is very expeditious without using toxic compounds such as phenol or chloroform. It can be used for phage identification and phage gene detection. PMID- 19188100 TI - Highly efficient and low-cost purification of lysozyme: a novel tris(hydroxymethyl)aminomethane immobilized affinity column. AB - A highly efficient and low-cost affinity chromatography strategy for lysozyme (LZM) purification is reported. Using tris(hydroxymethyl)aminomethane (Tris) as ligand and macroporous silica spheres as matrix, a novel affinity column was prepared. The high specificity, stability and repeatability of this Tris immobilized affinity column were proved by LZM separations from protein mixture solutions for 20 circles and 6 months test. LZM purified from chicken egg white on the Tris affinity column had even higher purity than the commercial standard and well-maintained activity of 8287 U/mg (activity of commercial LZM was 8171 U/mg). The efficient affinity process avoiding expensive or fragile ligand would bring advantages to the routine production of LZM from chicken egg white. PMID- 19188101 TI - Is BodyThink an efficacious body image and self-esteem program? A controlled evaluation with adolescents. AB - This study aimed to evaluate the efficacy of BodyThink, a widely disseminated body image and self-esteem program. Participants were 277, grade 7 students from 4 secondary schools in Australia. The intervention group (62 girls, 85 boys) participated in BodyThink during four 50-min lessons, while the control group (65 girls, 65 boys) received their usual classes. All participants completed baseline, postintervention and 3-month follow-up questionnaires. For girls, the intervention group reported higher media literacy and lower internalization of the thin ideal compared to the control group. For boys, the intervention group reported higher media literacy and body satisfaction than the control group. Although some positive outcomes were observed, it would be valuable to find ways to enhance the impact of BodyThink, especially in light of its wide dissemination. Suggestions for improving BodyThink are presented. PMID- 19188102 TI - Contributions of weight perceptions to weight loss attempts: differences by body mass index and gender. AB - Previous studies have consistently observed that women are more likely to perceive themselves as overweight compared to men. Similarly, women are more likely than men to report trying to lose weight. Less is known about the impact that self-perceived weight has on weight loss behaviors of adults and whether this association differs by gender. We conducted a cross-sectional analysis among an employee sample (n=899) to determine the association of self-perceived weight on evidence-based weight loss behaviors across genders, accounting for body mass index (BMI) and demographic characteristics. Women were more likely than men to consider themselves to be overweight across each BMI category, and were more likely to report attempting to lose weight. However, perceiving oneself to be overweight was a strong correlate for weight loss attempts across both genders. The effect of targeting accuracy of self-perceived weight status in weight loss interventions deserves research attention. PMID- 19188103 TI - Nucleation and growth of biomimetic apatite layers on 3D plotted biodegradable polymeric scaffolds: effect of static and dynamic coating conditions. AB - Apatite layers were grown on the surface of newly developed starch/polycaprolactone (SPCL)-based scaffolds by a 3D plotting technology. To produce the biomimetic coatings, a sodium silicate gel was used as nucleating agent, followed by immersion in a simulated body fluid (SBF) solution. After growing a stable apatite layer for 7 days, the scaffolds were placed in SBF under static, agitated (80 strokes min(-1)) and circulating flow perfusion (Q=4 ml min( 1); t(R)=15s) for up to 14 days. The materials were characterized by scanning electron microscopy/energy dispersive X-ray spectroscopy, Fourier transform infrared spectroscopy and thin-film X-ray diffraction. Cross-sections were obtained and the coating thickness was measured. The elemental composition of solution and coatings was monitored by inductively coupled plasma spectroscopy. After only 6 h of immersion in SBF it was possible to observe the formation of small nuclei of an amorphous calcium phosphate (ACP) layer. After subsequent SBF immersion from 7 to 14 days under static, agitated and circulating flow perfusion conditions, these layers grew into bone-like nanocrystalline carbonated apatites covering each scaffold fiber without compromising its initial morphology. No differences in the apatite composition/chemical structure were detectable between the coating conditions. In case of flow perfusion, the coating thickness was significantly higher. This condition, besides mimicking better the biological milieu, allowed for the coating of complex architectures at higher rates, which can greatly reduce the coating step. PMID- 19188104 TI - The effects of mega-dose ascorbic acid on skin flap survival: experimental study on rats. PMID- 19188105 TI - A nonlocal maximum likelihood estimation method for Rician noise reduction in MR images. AB - Postacquisition denoising of magnetic resonance (MR) images is of importance for clinical diagnosis and computerized analysis, such as tissue classification and segmentation. It has been shown that the noise in MR magnitude images follows a Rician distribution, which is signal-dependent when signal-to-noise ratio (SNR) is low. It is particularly difficult to remove the random fluctuations and bias introduced by Rician noise. The objective of this paper is to estimate the noise free signal from MR magnitude images. We model images as random fields and assume that pixels which have similar neighborhoods come from the same distribution. We propose a nonlocal maximum likelihood (NLML) estimation method for Rician noise reduction. Our method yields an optimal estimation result that is more accurate in recovering the true signal from Rician noise than NL means algorithm in the sense of SNR, contrast, and method error. We demonstrate that NLML performs better than the conventional local maximum likelihood (LML) estimation method in preserving and defining sharp tissue boundaries in terms of a well-defined sharpness metric while also having superior performance in method error. PMID- 19188106 TI - Algebraic decomposition of fat and water in MRI. AB - The decomposition of magnetic resonance imaging (MRI) data to generate water and fat images has several applications in medical imaging, including fat suppression and quantification of visceral fat. We introduce a novel algorithm to overcome some of the problems associated with current analytical and iterative decomposition schemes. In contrast to traditional analytical schemes, our approach is general enough to accommodate any uniform echo-shift pattern, any number of metabolites and signal samples. In contrast to region-growing method that use a smooth field-map initialization to resolve the ambiguities with the IDEAL algorithm, we propose to use an explicit smoothness constraint on the final field-map estimate. Towards this end, we estimate the number of feasible solutions at all the voxels, prior to the evaluation of the roots. This approach enables the algorithm to evaluate all the feasible roots, thus avoiding the convergence to the wrong solution. The estimation procedure is based on a modification of the harmonic retrieval (HR) framework to account for the chemical shift dependence in the frequencies. In contrast to the standard linear HR framework, we obtain the frequency shift as the common root of a set of quadratic equations. On most of the pixels with multiple feasible solutions, the correct solution can be identified by a simple sorting of the solutions. We use a region merging algorithm to resolve the remaining ambiguity and phase-wrapping. Experimental results indicate that the proposed algebraic scheme eliminates most of the difficulties with the current schemes, without compromising the noise performance. Moreover, the proposed algorithm is also computationally more efficient. PMID- 19188108 TI - Lossless compression of microarray images using image-dependent finite-context models. AB - The use of microarray expression data in state-of-the-art biology has been well established. The widespread adoption of this technology, coupled with the significant volume of data generated per experiment, in the form of images, has led to significant challenges in storage and query retrieval. In this paper, we present a lossless bitplane-based method for efficient compression of microarray images. This method is based on arithmetic coding driven by image-dependent multibitplane finite-context models. It produces an embedded bitstream that allows progressive, lossy-to-lossless decoding. We compare the compression efficiency of the proposed method with three image compression standards (JPEG2000, JPEG-LS, and JBIG) and also with the two most recent specialized methods for microarray image coding. The proposed method gives better results for all images of the test sets and confirms the effectiveness of bitplane-based methods and finite-context modeling for the lossless compression of microarray images. PMID- 19188107 TI - The validity of three-class Hotelling trace (3-HT) in describing three-class task performance: comparison of three-class volume under ROC surface (VUS) and 3-HT. AB - In order to describe multiclass classification performance, several figures of merit (FOM) have been proposed. Among the earliest and most widely known of these is the three-class Hotelling trace (3-HT). The goal of this paper is to present theoretical and empirical data demonstrating the failure of 3-HT as a measure of three-class task performance. To help do this, we contrast it to a newly proposed three-class FOM, the volume under the three-class receiver operating characteristic (ROC) surface (VUS). The VUS is obtained from a decision theory based three-class ROC analysis method which has been proved to extend the decision theoretic, linear discriminant analysis (LDA), and psychophysical foundations of binary ROC analysis to a three-class paradigm. We demonstrate empirically that the VUS and 3-HT do not have a monotonic relationship in general when describing three-class task performance. Numerical experiments demonstrated that the VUS provided reasonable results, while the 3-HT failed to distinguish between the case where all objects could be perfectly classified from the case where only one pair of the classes could be perfectly classified. We have provided theoretical explanations of this failure of 3-HT. The significance of this work goes beyond merely demonstrating the problems of the 3-HT, it demonstrates that a FOM that is mathematically correct and has a strong theoretical basis can provide results that violate a common sense understanding of three-class task performance. This fact raises the question of "how to evaluate a classification performance evaluation method?" We believe the answer to this question lies in the theoretical foundations of binary ROC analysis. We have thus contrasted the two FOMs in terms of three fundamental theories underlying binary ROC analysis: decision theory, binary linear discriminant analysis, and the equivalence of two psychophysical classification procedures. These theoretical investigations demonstrated the importance of extending and unifying all the fundamental theories of binary classification in the development of a three-class FOM; violating one of theses fundamental binary classification theories may, as it did for the L-HT, provide predictions of three-class task performance that do not agree with a common sense understanding of three-class task performance. PMID- 19188109 TI - Anatomy-guided lung lobe segmentation in X-ray CT images. AB - The human lungs are divided into five distinct anatomic compartments called the lobes, which are separated by the pulmonary fissures. The accurate identification of the fissures is of increasing importance in the early detection of pathologies, and in the regional functional analysis of the lungs. We have developed an automatic method for the segmentation and analysis of the fissures, based on the information provided by the segmentation and analysis of the airway and vascular trees. This information is used to provide a close initial approximation to the fissures, using a watershed transform on a distance map of the vasculature. In a further refinement step, this estimate is used to construct a region of interest (ROI) encompassing the fissures. The ROI is enhanced using a ridgeness measure, which is followed by a 3-D graph search to find the optimal surface within the ROI. We have also developed an automatic method to detect incomplete fissures, using a fast-marching based segmentation of a projection of the optimal surface. The detected incomplete fissure is used to extrapolate and smoothly complete the fissure. We evaluate the method by testing on data sets from normal subjects and subjects with mild to moderate emphysema. PMID- 19188110 TI - Lesion detection in dynamic FDG-PET using matched subspace detection. AB - We describe a matched subspace detection algorithm to assist in the detection of small tumors in dynamic positron emission tomography (PET) images. The algorithm is designed to differentiate tumors from background using the time activity curves (TACs) that characterize the uptake of PET tracers. TACs are modeled using linear subspaces with additive Gaussian noise. Using TACs from a primary tumor region of interest (ROI) and one or more background ROIs, each identified by a human observer, two linear subspaces are identified. Applying a matched subspace detector to these identified subspaces on a voxel-by-voxel basis throughout the dynamic image produces a test statistic at each voxel which on thresholding indicates potential locations of secondary or metastatic tumors. The detector is derived for three cases: using a single TAC with white noise of unknown variance, using a single TAC with known noise covariance, and detection using multiple TACs within a small ROI with known noise covariance. The noise covariance is estimated for the reconstructed image from the observed sinogram data. To evaluate the proposed method, a simulation-based receiver operating characteristic (ROC) study for dynamic PET tumor detection is designed. The detector uses a dynamic sequence of frame-by-frame 2-D reconstructions as input. We compare the performance of the subspace detectors with that of a Hotelling observer applied to a single frame image and of the Patlak method applied to the dynamic data. We also show examples of the application of each detection approach to clinical PET data from a breast cancer patient with metastatic disease. PMID- 19188111 TI - A model-based consecutive scanline tracking method for extracting vascular networks from 2-D digital subtraction angiograms. AB - We propose a new model-based algorithm for the automated tracking of vascular networks in 2-D digital subtraction angiograms. Consecutive scanline profiles are fitted by a parametric imaging model to estimate local vessel center point, radius, edge locations and direction. An adaptive tracking strategy is applied with appropriate termination criteria to track each vessel segment. When tracking stops, to prevent premature termination and to detect bifurcations, a look ahead detection scheme is used to search for possible continuation points of the same vessel segment or those of its bifurcated segments. The proposed algorithm can automatically extract the majority of the vascular network without human interaction other than initializing the start point and direction. Compared to other tracking methods, the proposed method highlights accurate estimation of local vessel geometry. Accurate geometric information and a hierarchical vessel network are obtained which can be used for further quantitative analysis of arterial networks to obtain flow conductance estimates. PMID- 19188112 TI - Suppression of metal artifacts in CT using a reconstruction procedure that combines MAP and projection completion. AB - Metal implants such as hip prostheses and dental fillings produce streak and star artifacts in the reconstructed computed tomography (CT) images. Due to these artifacts, the CT image may not be diagnostically usable. A new reconstruction procedure is proposed that reduces the streak artifacts and that might improve the diagnostic value of the CT images. The procedure starts with a maximum a posteriori (MAP) reconstruction using an iterative reconstruction algorithm and a multimodal prior. This produces an artifact-free constrained image. This constrained image is the basis for an image-based projection completion procedure. The algorithm was validated on simulations, phantom and patient data, and compared with other metal artifact reduction algorithms. PMID- 19188113 TI - Spatial characterization of FMRI activation maps using invariant 3-D moment descriptors. AB - A novel approach is proposed for quantitatively characterizing the spatial patterns of activation statistics in functional magnetic resonance imaging (fMRI) activation maps. Specifically, we propose using 3-D invariant moment descriptors, as opposed to the traditionally-employed magnitude-based features such as mean voxel statistics or percentage of activated voxels, to characterize the task specific spatial distribution of activation statistics within a given region of interest (ROI). The proposed method is applied to real fMRI data collected from 21 healthy subjects performing previously-learned right-handed finger tapping sequences that are either externally guided (EG) by a cue or internally guided (IG)--tasks expected to incur subtle differences in motor-related cortical and subcortical ROIs. Voxel-based activation statistics contrasting EG versus rest and IG versus rest are examined in multiple manually-drawn ROIs on unwarped brain images. Analyzing the activation statistics within each ROI using the proposed 3 D invariant moment descriptors detected significant group differences between the two tasks, thus quantitatively demonstrating that the spatial distribution of activation statistics within an ROI represent an important task-related attribute of brain activation. In contrast, conventional methods that solely rely on activation statistic magnitudes and disregard spatial information showed reduced discriminability. Normally, incorporating spatial information would merely increase inter-subject variability partly due to differences in brain size and subject's orientation in the scanner. Yet, our results suggest that the proposed spatial features, which are invariant to similarity transformations, can effectively account for such inter-subject variability, while enhancing the sensitivity in detecting task-specific activation. Thus, we argue that this novel quantitative description of the "3-D texture" of activation maps provides new directions to explore for ROI-based fMRI analysis. PMID- 19188114 TI - Deterministic and probabilistic tractography based on complex fibre orientation distributions. AB - We propose an integral concept for tractography to describe crossing and splitting fibre bundles based on the fibre orientation distribution function (ODF) estimated from high angular resolution diffusion imaging (HARDI). We show that in order to perform accurate probabilistic tractography, one needs to use a fibre ODF estimation and not the diffusion ODF. We use a new fibre ODF estimation obtained from a sharpening deconvolution transform (SDT) of the diffusion ODF reconstructed from q-ball imaging (QBI). This SDT provides new insight into the relationship between the HARDI signal, the diffusion ODF, and the fibre ODF. We demonstrate that the SDT agrees with classical spherical deconvolution and improves the angular resolution of QBI. Another important contribution of this paper is the development of new deterministic and new probabilistic tractography algorithms using the full multidirectional information obtained through use of the fibre ODF. An extensive comparison study is performed on human brain datasets comparing our new deterministic and probabilistic tracking algorithms in complex fibre crossing regions. Finally, as an application of our new probabilistic tracking, we quantify the reconstruction of transcallosal fibres intersecting with the corona radiata and the superior longitudinal fasciculus in a group of eight subjects. Most current diffusion tensor imaging (DTI)-based methods neglect these fibres, which might lead to incorrect interpretations of brain functions. PMID- 19188115 TI - Likelihood-based hypothesis tests for brain activation detection from MRI data disturbed by colored noise: a simulation study. AB - Functional magnetic resonance imaging (fMRI) data that are corrupted by temporally colored noise are generally preprocessed (i.e., prewhitened or precolored) prior to functional activation detection. In this paper, we propose likelihood-based hypothesis tests that account for colored noise directly within the framework of functional activation detection. Three likelihood-based tests are proposed: the generalized likelihood ratio (GLR) test, the Wald test, and the Rao test. The fMRI time series is modeled as a linear regression model, where one regressor describes the task-related hemodynamic response, one regressor accounts for a constant baseline and one regressor describes potential drift. The temporal correlation structure of the noise is modeled as an autoregressive (AR) model. The order of the AR model is determined from practical null data sets using Akaike's information criterion (with penalty factor 3) as order selection criterion. The tests proposed are based on exact expressions for the likelihood function of the data. Using Monte Carlo simulation experiments, the performance of the proposed tests is evaluated in terms of detection rate and false alarm rate properties and compared to the current general linear model (GLM) test, which estimates the coloring of the noise in a separate step. Results show that theoretical asymptotic distributions of the GLM, GLR, and Wald test statistics cannot be reliably used for computing thresholds for activation detection from finite length time series. Furthermore, it is shown that, for a fixed false alarm rate, the detection rate of the proposed GLR test statistic is slightly, but (statistically) significantly improved compared to that of the common GLM-based tests. Finally, simulations results reveal that all tests considered show seriously inferior performance if the order of the AR model is not chosen sufficiently high to give an adequate description of the correlation structure of the noise, whereas the effects of (slightly) overmodeling are observed to be less harmful. PMID- 19188116 TI - An EM approach to MAP solution of segmenting tissue mixtures: a numerical analysis. AB - This work presents an iterative expectation-maximization (EM) approach to the maximum a posteriori (MAP) solution of segmenting tissue mixtures inside each image voxel. Each tissue type is assumed to follow a normal distribution across the field-of-view (FOV). Furthermore, all tissue types are assumed to be independent from each other. Under these assumptions, the summation of all tissue mixtures inside each voxel leads to the image density mean value at that voxel. The summation of all the tissue mixtures' unobservable random processes leads to the observed image density at that voxel, and the observed image density value also follows a normal distribution (image data are observed to follow a normal distribution in many applications). By modeling the underlying tissue distributions as a Markov random field across the FOV, the conditional expectation of the posteriori distribution of the tissue mixtures inside each voxel is determined, given the observed image data and the current-iteration estimation of the tissue mixtures. Estimation of the tissue mixtures at next iteration is computed by maximizing the conditional expectation. The iterative EM approach to a MAP solution is achieved by a finite number of iterations and reasonable initial estimate. This MAP-EM framework provides a theoretical solution to the partial volume effect, which has been a major cause of quantitative imprecision in medical image processing. Numerical analysis demonstrated its potential to estimate tissue mixtures accurately and efficiently. PMID- 19188117 TI - Linear superposition electrical impedance tomography imaging with multiple electrical/biopsy probes. AB - In medical diagnostics, tissue is often examined with multiple discrete biopsies taken under ultrasound placement. In a previous theoretical study, we have suggested that the linear nature of the equations used in electrical impedance tomography (EIT) can be employed with the conventional practice of biopsy sampling to produce an image of the tissue between the biopsy samplings. Specifically, the biopsy probes can be used to record EIT-type electrical data during the discrete tissue sampling. The location of the discrete biopsy needle insertions available from the ultrasound placement of the probes can be combined with the electrical measurement data and used with linear superposition to produce a complete EIT image of the tissue between the sampled sites. In this study, we explore the concept experimentally using gel phantoms to simulate tissue and heterogeneities in the tissue. The experiments are performed in 2-D and 3-D configurations, and data are taken discretely, one at a time, through single electrical probe insertions. In the 2-D configuration, we were able to produce images of reasonable quality for heterogeneities with a diameter larger than 3 mm (conductivity ratio 1:5) and with relative conductivity differences above 50% (diameter 5 mm). PMID- 19188118 TI - Nonlinear stochastic regularization to characterize tissue residue function in bolus-tracking MRI: assessment and comparison with SVD, block-circulant SVD, and Tikhonov. AB - An accurate characterization of tissue residue function R(t) in bolus-tracking magnetic resonance imaging is of crucial importance to quantify cerebral hemodynamics. R(t) estimation requires to solve a deconvolution problem. The most popular deconvolution method is singular value decomposition (SVD). However, SVD is known to bear some limitations, e.g., R(t) profiles exhibit nonphysiological oscillations and take on negative values. In addition, SVD estimates are biased in presence of bolus delay and dispersion. Recently, other deconvolution methods have been proposed, in particular block-circulant SVD (cSVD) and Tikhonov regularization (TIKH). Here we propose a new method based on nonlinear stochastic regularization (NSR). NSR is tested on simulated data and compared with SVD, cSVD, and TIKH in presence and absence of bolus dispersion. A clinical case in one patient has also been considered. NSR is shown to perform better than SVD, cSVD, and TIKH in reconstructing both the peak and the residue function, in particular when bolus dispersion is considered. In addition, differently from SVD, cSVD, and TIKH, NSR always provides positive and smooth R(t). PMID- 19188119 TI - A multitask learning model for online pattern recognition. AB - This paper presents a new learning algorithm for multitask pattern recognition (MTPR) problems. We consider learning multiple multiclass classification tasks online where no information is ever provided about the task category of a training example. The algorithm thus needs an automated task recognition capability to properly learn the different classification tasks. The learning mode is "online" where training examples for different tasks are mixed in a random fashion and given sequentially one after another. We assume that the classification tasks are related to each other and that both the tasks and their training examples appear in random during "online training." Thus, the learning algorithm has to continually switch from learning one task to another whenever the training examples change to a different task. This also implies that the learning algorithm has to detect task changes automatically and utilize knowledge of previous tasks for learning new tasks fast. The performance of the algorithm is evaluated for ten MTPR problems using five University of California at Irvine (UCI) data sets. The experiments verify that the proposed algorithm can indeed acquire and accumulate task knowledge and that the transfer of knowledge from tasks already learned enhances the speed of knowledge acquisition on new tasks and the final classification accuracy. In addition, the task categorization accuracy is greatly improved for all MTPR problems by introducing the reorganization process even if the presentation order of class training examples is fairly biased. PMID- 19188120 TI - Sampled-data adaptive NN tracking control of uncertain nonlinear systems. AB - In this paper, for a class of single-input-single-output (SISO) uncertain nonlinear systems, adaptive neural tracking controllers designed for digital computer implementation are proposed. The overall scheme can be considered as a sampled-data adaptive neural control system. As an intermediate result, it is proven that, for a sufficiently small sampling period, the emulated adaptive neural controller i.e., the discrete implementation of the continuous-time adaptive neural network controller ensures semiglobal uniformly ultimate boundedness of the closed-loop system. Then, based on the exact discrete-time model, a controller redesign is proposed that performs efficiently for sampling periods for which the emulation controller fails. The redesigned controller consists of two terms: the emulated control law and an extra robustness term designed to increase the order of the perturbation (with respect to the sampling period) in the Lyapunov difference. In all cases, high-order neural networks are employed to approximate the unknown nonlinearities. Using Lyapunov techniques, it is proven that, for a sufficiently small sampling period, the proposed redesigned controller ensures the (semiglobal) boundedness of all the signals in the closed loop while the output of the system converges to a small neighborhood of the desired trajectory. Simulation results illustrate the superiority of the proposed scheme with respect to the emulation controller and verify the theoretical analysis. PMID- 19188121 TI - Geometry-driven distributed compression of the plenoptic function: performance bounds and constructive algorithms. AB - In this paper, we study the sampling and the distributed compression of the data acquired by a camera sensor network. The effective design of these sampling and compression schemes requires, however, the understanding of the structure of the acquired data. To this end, we show that the a priori knowledge of the configuration of the camera sensor network can lead to an effective estimation of such structure and to the design of effective distributed compression algorithms. For idealized scenarios, we derive the fundamental performance bounds of a camera sensor network and clarify the connection between sampling and distributed compression. We then present a distributed compression algorithm that takes advantage of the structure of the data and that outperforms independent compression algorithms on real multiview images. PMID- 19188122 TI - Geometry-based demosaicking. AB - Demosaicking is a particular case of interpolation problems where, from a scalar image in which each pixel has either the red, the green or the blue component, we want to interpolate the full-color image. State-of-the-art demosaicking algorithms perform interpolation along edges, but these edges are estimated locally. We propose a level-set-based geometric method to estimate image edges, inspired by the image inpainting literature. This method has a time complexity of O(S) , where S is the number of pixels in the image, and compares favorably with the state-of-the-art algorithms both visually and in most relevant image quality measures. PMID- 19188123 TI - Pencil back-projection method for SAR imaging. AB - We present a high-resolution method for spotlight mode SAR imaging that utilizes parametric modeling of projected target reflectivity density function and tomographic reconstruction. The method requires no polar-to-cartesian interpolation in spectral domain. Utilization of forward-backward total least squares bandpass matrix pencil method allows super resolution to be achieved in range for a single imaging angle. Hence, the quality of the image reconstructed by convolution back-projection is also improved. It is shown that the method is very resistant to noise and can generate images down to very low SNR values. Direct formulation in terms of physical quantities such as electric field and current density is another contribution of this paper. PMID- 19188124 TI - A fast multilevel algorithm for wavelet-regularized image restoration. AB - We present a multilevel extension of the popular "thresholded Landweber" algorithm for wavelet-regularized image restoration that yields an order of magnitude speed improvement over the standard fixed-scale implementation. The method is generic and targeted towards large-scale linear inverse problems, such as 3-D deconvolution microscopy. The algorithm is derived within the framework of bound optimization. The key idea is to successively update the coefficients in the various wavelet channels using fixed, subband-adapted iteration parameters (step sizes and threshold levels). The optimization problem is solved efficiently via a proper chaining of basic iteration modules. The higher level description of the algorithm is similar to that of a multigrid solver for PDEs, but there is one fundamental difference: the latter iterates though a sequence of multiresolution versions of the original problem, while, in our case, we cycle through the wavelet subspaces corresponding to the difference between successive approximations. This strategy is motivated by the special structure of the problem and the preconditioning properties of the wavelet representation. We establish that the solution of the restoration problem corresponds to a fixed point of our multilevel optimizer. We also provide experimental evidence that the improvement in convergence rate is essentially determined by the (unconstrained) linear part of the algorithm, irrespective of the type of wavelet. Finally, we illustrate the technique with some image deconvolution examples, including some real 3-D fluorescence microscopy data. PMID- 19188125 TI - Robust and efficient feature tracking for indoor navigation. AB - Robust feature tracking is a requirement for many computer vision tasks such as indoor robot navigation. However, indoor scenes are characterized by poorly localizable features. As a result, indoor feature tracking without artificial markers is challenging and remains an attractive problem. We propose to solve this problem by constraining the locations of a large number of nondistinctive features by several planar homographies which are strategically computed using distinctive features. We experimentally show the need for multiple homographies and propose an illumination-invariant local-optimization scheme for motion refinement. The use of a large number of nondistinctive features within the constraints imposed by planar homographies allows us to gain robustness. Also, the lesser computation cost in estimating these nondistinctive features helps to maintain the efficiency of the proposed method. Our local-optimization scheme produces subpixel accurate feature motion. As a result, we are able to achieve robust and accurate feature tracking. PMID- 19188126 TI - Software engineering and scale-free networks. AB - Complex-network theory is a new approach in studying different types of large systems in both the physical and the abstract worlds. In this paper, we have studied two kinds of network from software engineering: the component dependence network and the sorting comparison network (SCN). It is found that they both show the same scale-free property under certain conditions as complex networks in other fields. These results suggest that complex-network theory can be a useful approach to the study of software systems. The special properties of SCNs provide a more repeatable and deterministic way to study the evolution and optimization of complex networks. They also suggest that the closer a sorting algorithm is to the theoretical optimal limit, the more its SCN is like a scale-free network. This may also indicate that, to store and retrieve information efficiently, a concept network might need to be scale-free. PMID- 19188127 TI - [Is Turkish bath water potable?: The baths of Sidi-Bel-Abbes]. AB - In Algeria, large numbers of people regularly go to Turkish baths or "Hammams". The cold tap water of the baths in the town of Sidi-Bel-Abbes (Algeria) comes either from wells or from a mixture of potable waterworks water and well water. Its principal use is for personal hygiene (washing). However, the steam heat generates thirst that can cause users to drink cold water during the steam bath. In addition, the wells feeding the baths are often poorly protected and especially badly treated. To ascertain whether their water quality, particularly bacteriological, meets the requirements for drinking water, we studied the characteristics of water from ten Turkish baths in Sidi-Bel-Abbes. Bacteriological analyses of cold water showed signs of contamination of fecal origin in 50% of the samples analysed. Moreover two water points from two of the baths appeared to have permanent fecal contamination. The physicochemical analysis showed that the water was very high in calcium (up to 550 mg/L) and magnesium (up to 299 mg/L). The maximum nitrate level observed was 68 mg/L. This study thus showed the existence of a health risk due to deterioration in the quality of the bath water and demonstrated the need for protection of the wells, frequent purification, and regular microbiological testing. PMID- 19188128 TI - [Current bacterial causes of osteomyelitis in children with sickle cell disease]. AB - INTRODUCTION: Osteomyelitis in children with sickle cell disease has been caused mainly by salmonella species, but in recent years, greater variation has resulted in other bacteria sometimes heading this list. We conducted this survey to determine the present distribution of causative bacteria, to improve initial probabilistic antibiotic therapy. MATERIAL AND METHODS: This retrospective survey reviewed files of children hospitalised during the 8-year period from 1998 to 2005 for osteomyelitis who had haemoglobinopathy SS or SC. In all, we identified 32 patients (mean age: 6.5 years) - 22 boys and 10 girls, 18 with SS and 14 SC. Eleven children with haemoglobinopathy AS were not included. Thirteen children had acute osteomyelitis and 19 chronic osteomyelitis. Bacteriological research included blood cultures, needle aspiration or biopsy, and cytobacteriological examination of pus. RESULTS: Of 4 blood cultures, one was positive. Only one needle aspiration was performed, and it was positive. Pus was examined in 19 cases and positive in 16 (84%). No bacteriological research was performed in 8 of the cases (25%) of acute osteomyelitis. Staphylococcus accounted for 22% of the bacteria identified, salmonella for 19% and Pseudomonas aeruginosa for 6%. CONCLUSION: Initial probabilistic antibiotic therapy for osteomyelitis in children with sickle cell disease must target staphylococcus and salmonella, preferably by combining a third-generation cephalosporin and aminoglycoside. Systematic bacteriological testing will provide a more reliable selection and improve initial treatment. PMID- 19188129 TI - [Pediatric management of sickle cell disease: experience at the Charles de Gaulle University Children's Hospital in Ouagadougou (Burkina Faso)]. AB - Sickle cell disease is a genetic disease most common in blacks. We retrospectively collected records for patients with sickle cell disease who were seen from January 2002 through September 2006 to assess the care provided for this disease at Charles de Gaulle University Children's Hospital of Ouagadougou. In all, 88 patients were monitored quarterly at outpatient visits for sickle cell disease, in the absence of any crisis. Their age ranged from 6 months to 16 years, with an average age of 7. There were more boys than girls, with a sex ratio of 1.44. The distribution according to sickle cell genotype showed that SC accounted for 62% of cases, while SS forms were more frequent until the age of 5. All children have received the immunizations in the standard Expanded Programme on Immunization (EPI) [diphtheria, tetanus, pertussis, polio, measles and yellow fever]. The immunization rates for non-EPI vaccines including hepatitis B, Haemophilus influenzae B, Salmonella typhi, meningitis, pneumonia and the combined vaccine against measles, mumps and rubella ranged from 94 to 100%. A prophylactic anti-anaemic agent was made with folic acid often associated with iron. In addition, patients receive malaria chemoprophylaxis. Chloroquine was initially provided, and since 2006, children have been receiving sulfadoxine pyrimethamine. Our encouraging results deserve reinforcement in the short-term - at the local level by neonatal screening, the creation of an immunization unit, and the systematization of antibiotic prophylaxis, and in the medium-term by implementation of a National sickle cell disease programme to help meet the objective of a 40% reduction in mortality among affected children younger than 5 years by 2015, set by the Sickle Cell Disease International Organization. PMID- 19188130 TI - [Pelvic actinomycosis in Tunisia: five cases]. AB - Actinomycosis is a rare suppurative disease due to Actinomyces species. These Gram-positive, non-acid fast anaerobic filamentous bacteria are normal inhabitants of the human body, tending to reside in the oropharynx and bowel but are occasionally found in the vagina. Pelvic actinomycosis is a rare bacterial disease in women. Clinical manifestations are various and non specific and may be acute or chronic. No consensus exists for treatment. We reviewed files and identified all five cases of pelvic actinomycosis managed at Obstetrics and Gynaecology department "A" at the Maternity Center of Tunis over an eight-year period (1998-2005). The women's average age was 39.2 years. One patient was menopausal and consulted for bleeding. The other four patients were younger and had all been using an intrauterine device (IUD) for contraception. They presented with acute clinical manifestations. Their main symptom was pelvic pain. Three women had fever, and two presented with urinary tract obstruction. All patients had surgery. A pelvic abscess was found in four cases. Laparoscopic management was possible in only one case. Laparotomy was necessary in the other four. Four women had adnexectomies, two with hysterectomy. Digestive complications occurred in three cases. Actinomycosis was diagnosed only after surgery, by the histological examination. This series confirms the difficulties encountered in the management of pelvic actinomycosis. We review the recent literature and describe the diagnostic and therapeutic procedures currently recommended. The relationship between pelvic actinomycosis and IUDs, the most common method of contraception in Tunisia, is clearly established. Clinical diagnosis of pelvic actinomycosis is difficult because the symptoms are non-specific. Laboratory tests can help by showing serious inflammation, however. Imaging findings are also non-specific and may suggest an abscess or an inflammatory or neoplastic process. Interventional radiology, specifically CT- or ultrasound-guided aspiration biopsy can facilitate diagnosis, which in any case requires a histological examination. Treatment of pelvic actinomycosis is not standardized and depends on clinical form. Medical treatment is based on long-term penicillin G. It is always necessary and may be used alone in cases of preoperative diagnosis. Surgery should be offered only in resistant cases and should always be followed by long-term antibiotic treatment. The prognosis of correctly treated pelvic actinomycosis is generally good. PMID- 19188131 TI - [Imperforate hymen: Can it be treated without damaging the hymenal structure?]. AB - Imperforate hymen is a rare congenital anomaly, with an incidence of about 1 in 2000 female births. It is generally diagnosed during puberty. Treatment generally consists of a hymenotomy or a hymenectomy. Because the hymen is a symbol of virginity in some communities, its destruction can be source of social problems for some girls. OBJECTIVES: We discuss the diagnostic but especially therapeutic aspects of imperforate hymens and possible surgical techniques, in particular those that preserve the hymen. MATERIAL AND METHODS: We describe the cases of 5 girls treated in our department for imperforate hymen between 2001 and 2007. Two of them required the safeguarding of the normal architecture of their hymen to preserve the appearance of virginity. We analysed diagnostic features and surgical techniques. RESULTS: The average age of our patients was 14.8 years (range: 11 and 17 years). The most frequent reason for consultation was pelvic pain with primary amenorrhea. Inspection of the vulva revealed in all cases a dome-shaped purplish-red hymeneal membrane. Hymeneal incision allowed drainage of old previously blocked menstrual blood. Three patients were treated by radial incisions of the hymen. The parents of 2 patients demanded that their hymens be preserved. Accordingly, one had a simple excision of a central flange of the hymen and the other was treated by a similar technique that also used a Foley catheter . All five patients did well after surgical treatment. The techniques used to preserve the hymen resulted in an apparently intact annular hymen. CONCLUSION: Imperforate hymen is a rare anomaly. Its diagnosis is simple. The traditional technique of radial incisions is a simple procedure that yields good results. The technique using the Foley catheter is an adequate alternative when preservation of the hymen is required. PMID- 19188133 TI - [National coordination of the ambulatory treatment centers (ATC) in Gabon: a new process to conduct the scaling up of care for people living with HIV-AIDS]. AB - Gabonese authorities are strongly mobilized in the fight against AIDS. With a national seroprevalence of 5.9%, 54,000 people are living with HIV-AIDS. Starting from the experience conducted on three ambulatory treatment centers (ATC) [Libreville, Franceville, Port-Gentil] in collaboration with the French Red Cross, Gabonese authorities decided the scaling up of HIV patients' care to seven new ATC at a national level. The increasing number of structures conducted to standardize treatment's guidelines, training of caregivers, quality of care, coordination, monitoring and evaluation. Technical and medical supervision of the ten ATC were attributed to the French Red Cross by Gabonese authorities with the financial support of the French Agency for Development. The ten ATC, founded by the ministry of public health, were then organized within a network under the responsibility of a national coordination. This structure created in September 2007 represents the main organism to conduct, evaluate and follow-up activities and functioning of the ten ATC. All these activities are conducted in agreement with the national program of fight against AIDS. Within one year this structure of coordination allowed to organize the training of 208 caregivers, to elaborate a team of national and international experts, to start a process of national guidelines, to elaborate technical procedures and indicators for monitoring, follow-up and evaluation. In June 2007, 5 ATC were in function taking care of 7,062 PLWA. In November 2008, 9 ATC were in function taking care of 8,174 PLWA. This project is planned for four years. It might allow to structure and organize a national network of care for PLWA according to the national strategy. This procedure of scaling up under the responsibility of a national team of technical and operational coordination is a new process. It completes and strengthens the national organization process. PMID- 19188132 TI - [Ten years of commitment to persons living with HIV-AIDS: evaluation of the management in three ambulatory treatment centers of the French Red Cross in Africa]. AB - The French Red Cross (FRC) has developed a strategy for the follow-up of people living with HIV-AIDS in Africa by setting-up and managing healthcare centers specialized in the management of HIV infection. Nearly one hundred and seventy thousand patients have had access to care in fifteen Ambulatory Treatment Centers (ATCs). For the ten years anniversary of the FRC's AIDS programs, we present the results of the evaluation of three ATCs in Africa. These results which show a low mortality rate in the patients on antiretroviral therapy and a very low rate of second line patients (1.5%) confirm the efficiency of the management of people living with HIV in the ATCs and generally of the AIDS programs of the FRC. However, the mortality remains high in patients who have a CD4+ cell count of less than 50/mm3 when antiretrovirals (ARVs) initiation. Services and care dispensed in the ATCs and particularly the antiretroviral therapy have demonstrated their feasibility and efficiency. In ten years, the challenge of the management of HIV has changed. Today, there is a need to integrate additional actions in the area of the supply of primary healthcare, of training and of motivation of the care providers. In addition, the follow-up/evaluation of the patient management programs remains useful to determine the impact and long-term efficacy of ARVs in resource-limited countries. PMID- 19188134 TI - EGFR, pMAPK, pAkt and PTEN status by immunohistochemistry: correlation with clinical outcome in HER2-positive metastatic breast cancer patients treated with trastuzumab. AB - BACKGROUND: In an attempt to identify markers of resistance to trastuzumab, we evaluated both the profiling of human epidermal growth factor receptor 2 (HER2) positive tumor cells measuring the relative levels of EGFR, pMAPK, pAkt and PTEN and their correlations with clinical outcome in HER2-positive metastatic breast cancer patients treated with trastuzumab. PATIENTS AND METHODS: Tumor tissues for this retrospective analysis were available from 45 out of 76 patients with metastatic breast cancer treated from April 1999 to March 2006 with trastuzumab based therapy at our Institution. Evaluations of EGFR, pMAPK, pAkt and PTEN status by immunohistochemistry (IHC) were carried out on all 45 tissue samples and their correlations with response to trastuzumab, incidence of central nervous system (CNS) metastases, time to progression (TTP), overall survival from diagnosis of breast cancer (OS1), from diagnosis of metastatic disease (OS2) and from the start of trastuzumab (OS3) were analyzed. RESULTS: We observed that TTP (P = 0.001) and median OS2 and OS3 were significantly longer in patients responsive to trastuzumab-based regimen compared with nonresponsive patients. EGFR, pMAPK, pAkt and PTEN status by IHC were not significantly associated with response to trastuzumab, TTP, overall survival (OS1, OS2, OS3) and CNS metastases incidence. A trend for shorter OS3 was observed for pMAPK-positive patients compared with pMAPK-negative patients (22.8 versus 31.2 months; P = 0.076). Median OS1 resulted shorter in 22 pAkt-positive patients (69.8 months) compared with 23 pAkt-negative patients (108.2 months); P = 0.091. It is likely that high expression of pMAPK (pMAPK-positive status) or pAkt (pAkt-positive status) could identify a subgroup of HER2-positive tumors with high activity of proliferation and survival pathways and with resistance to trastuzumab. CONCLUSIONS: In HER2 positive metastatic breast cancers, EGFR, pMAPK, pAkt and PTEN status evaluated by IHC was not significantly associated with response to trastuzumab, TTP, OS and CNS metastases incidence. However, HER2 status determined by IHC and/or FISH assays may not be sufficient to predict response to trastuzumab-based therapy. PMID- 19188135 TI - Osteonecrosis of the jaw under bisphosphonate and antiangiogenic therapies: cumulative toxicity profile? PMID- 19188136 TI - A randomized, phase II trial of two dose schedules of carboplatin/paclitaxel/cetuximab in stage IIIB/IV non-small-cell lung cancer (NSCLC). AB - BACKGROUND: This trial investigated the efficacy and safety of weekly cetuximab combined with two different schedules of paclitaxel/carboplatin for stage IIIB/IV non-small-cell lung cancer (NSCLC). METHODS: A total of 168 patients with previously untreated stage IIIB/IV NSCLC were randomized to arm A, cetuximab (400 mg/m(2) day 1 followed by weekly 250 mg/m(2)) + paclitaxel (Taxol) (225 mg/m(2))/carboplatin (AUC6) day 1 every 3 weeks or arm B, same cetuximab regimen plus paclitaxel (100 mg/m(2)) days 1, 8, and 15 every 3 weeks and carboplatin (AUC6) day 1 every 4 weeks. Treatment continued for a four-cycle maximum. Patients with a complete response, partial response, or stable disease after four cycles could receive cetuximab 250 mg/m(2)/week until disease progression or unacceptable toxicity. The primary end point was to evaluate progression-free survival (PFS). RESULTS: Median PFS was 4.7 and 4.3 months for arms A and B, respectively (6-month PFS, 27.3% versus 30.9%). Median overall survival was 11.4 versus 9.8 months for arms A and B, respectively; estimated 1-year survival, 47.7% versus 39.3%; and objective response rate, 29.6% versus 25%. The regimen was well tolerated with rash and hematologic toxicity being most common. CONCLUSIONS: This study did not meet the prespecified benchmark of 35% 6-month PFS rate; both combination schedules of cetuximab plus paclitaxel/carboplatin were feasible and equivalent for treating advanced NSCLC. PMID- 19188137 TI - Protein classification combining surface analysis and primary structure. AB - In this work, we propose a method for protein classification that combines the features extracted from both the primary structure and the surface analysis of a given protein. The surface analysis is used to find the amino acids that belong to the surface of the proteins. The most important finding of this work is to show that the features extracted from the amino acids that belong to the surface are useful in the classification process, since their contribution is partially independent from that of the features extracted from the whole primary structure; this property is used to build an ensemble of classifiers. The experimental results demonstrate the effectiveness of the proposed system. The idea is validated using three different data sets and three different feature extraction methods: 2-gram; residue couple; pseudo amino acid composition. PMID- 19188138 TI - Stabilization and humanization of a single-chain Fv antibody fragment specific for human lymphocyte antigen CD19 by designed point mutations and CDR-grafting onto a human framework. AB - A single-chain Fv (scFv) fragment derived from the murine antibody 4G7, specific for human lymphocyte CD19, was engineered for stability and expression in Escherichia coli in view of future use as a therapeutic protein. We compared two orthogonal knowledge-based procedures. In one approach, we designed a mutant with 14 single amino-acid substitutions predicted to correct destabilizing residues in the 4G7-wt sequence to create 4G7-mut. In the second variant, the murine CDRs were grafted to the human acceptor framework huVkappa3-huV(H)3, with 11 additional point mutations introduced to obtain a better match between CDR graft and acceptor framework, to arrive at 4G7-graft. Compared to 4G7-wt, 4G7-mut showed greater thermodynamic stability in guanidinium chloride-induced equilibrium denaturation experiments and somewhat greater stability in human serum. The loop graft maintained the comparatively high stability of the murine loop donor, but did not improve it further. Our analysis indicates that this is due to subtle strain introduced between CDRs and framework, mitigating the otherwise highly favorable properties of the human acceptor framework. This slight strain in the loop graft is also reflected in the binding affinities for CD19 on leukemic cells of 8.4 nM for 4G7-wt, 16.4 nM for 4G7-mut and 30.0 nM for 4G7-graft. This comparison of knowledge-based mutation and loop-grafting-based approaches will be important, when moving molecules forward to therapeutic applications. PMID- 19188139 TI - Response to: Overweight and obesity among adolescents in Norway: a response from the UK. PMID- 19188140 TI - Overweight and obesity among adolescents in Norway: a response from the UK. PMID- 19188141 TI - International benchmarking. Option or illusion? PMID- 19188142 TI - Overcoming cancer immune tolerance and escape. AB - Although HER2/neu-targeted cancer vaccines have shown initial promise in the adjuvant setting, a therapeutic vaccine remains elusive due to the tumor escape mechanisms of established cancer. As described by Seavey et al. in this issue of CCR, a Listeria-delivered vaccine may help overcome immune tolerance, leading to an effective therapeutic vaccine. PMID- 19188143 TI - A new fork for clinical application: targeting forkhead transcription factors in cancer. AB - Forkhead O transcription factors (FOXO) play a pivotal role in the regulation of a myriad of cellular functions including cell cycle arrest, cell death, and protection from stress stimuli. Activation of cell survival pathways such as phosphoinositide-3-kinase/AKT/IKK or RAS/mitogen-activated protein kinase are known to phosphorylate FOXOs at different sites which cause FOXOs nuclear exclusion and degradation, resulting in the suppression of FOXO's transcriptional activity. Perturbation of FOXO's function leads to deregulated cell proliferation and accumulation of DNA damage, resulting in diseases such as cancer. Emerging evidence shows that active FOXO proteins are crucial for keeping cells in check; and inactivation of FOXO proteins is associated with tumorigenesis, including breast cancer, prostate cancer, glioblastoma, rhabdomyosarcoma, and leukemia. Moreover, clinically used drugs like paclitaxel, imatinib, and doxorubicin have been shown to achieve their therapeutic effects through activation of FOXO3a and FOXO3a targets. In this review, we will focus the novel functions of FOXOs revealed in recent studies and further highlight FOXOs as new therapeutic targets in a broad spectrum of cancers. PMID- 19188145 TI - An interstitial deletion at 3p21.3 results in the genetic fusion of MLH1 and ITGA9 in a Lynch syndrome family. AB - PURPOSE: Germline mutations in DNA mismatch repair genes, mainly MLH1 or MSH2, have been shown to predispose with high penetrance for the development of the clinical phenotype of hereditary nonpolyposis colorectal cancer (Lynch syndrome). Here, we describe the discovery and first functional characterization of a novel germline MLH1 mutant allele. EXPERIMENTAL DESIGN: A large kindred including 54 potential carriers was investigated at the molecular level by using different types of PCR experiments, gene cloning, transfection studies, Western blot experiments, and mismatch repair assays to identify and characterize a novel MLH1 mutant allele. Twenty-two of 54 putative carriers developed colon cancer or other tumors, including breast cancer. RESULTS: The identified MLH1 mutant allele emerged from an interstitial deletion on chromosome 3p21.3, leading to an in frame fusion of MLH1 (exons 1-11) with ITGA9 (integrin alpha 9; exons 17-28). The deleted area has a size of about 400 kb; codes for LRRFIP2 (leucine-rich repeat in flightless interaction protein 2), GOLGA4 (Golgi autoantigen, golgin subfamily a, 4), and C3orf35/APRG1 (chromosome 3 open reading frame 35/AP20 region protein 1); and partly disrupts the AP20 region implicated in major epithelial malignancies. Tumor cells lost their second MLH1 allele. The MLH1*ITGA9 fusion protein provides no capability for DNA mismatch repair. Murine fibroblasts, expressing a doxycycline-inducible MLH1*ITGA9 fusion gene, exhibit a loss-of contact inhibition phenotype. CONCLUSIONS: This is the first description of a functional gene fusion of the human MLH1 gene, resulting in the loss of mismatch repair capabilities. The MLH1*ITGA9 fusion allele, together with deletions of the AP20 region, presumably defines a novel subclass of Lynch syndrome patients, which results in an extended tumor spectrum known from hereditary nonpolyposis colorectal cancer and Muir-Torre syndrome patients. PMID- 19188144 TI - Combination chemoprevention for colon cancer targeting polyamine synthesis and inflammation. AB - Increased polyamine synthesis and inflammation have long been associated with colon carcinogenesis in both preclinical models and in humans. Recent experimental studies suggest that polyamines may be mechanistically involved in colonic inflammatory processes. Genetic epidemiology results indicate that a single nucleotide polymorphism influencing the expression of a polyamine biosynthetic gene is associated with both risk of colon polyp occurrence and recurrence, and the response to aspirin as a polyp preventive agent. A prospective, randomized, placebo-controlled clinical trial of combination difluoromethylornithine, a selective inhibitor of polyamine synthesis, and sulindac, a nonsteroidal anti-inflammatory drug, found that the 3-year treatment was associated with a 70% reduction of recurrence of all adenomas, and over a 90% reduction of recurrence of advanced and/or multiple adenomas, without evidence of serious toxicities. This proof-of-principle trial indicates that targeting polyamine synthesis and inflammation can be an effective strategy for preventing the occurrence of the advanced and/or multiple adenomas that are most closely associated with the development of colon cancers in humans. PMID- 19188146 TI - Functional B7.2 and B7-H2 molecules on myeloma cells are associated with a growth advantage. AB - PURPOSE: B7 family molecules expressed on antigen-presenting cells stimulate or inhibit normal immune responses. The aim of this study was to investigate whether functional B7.2 and B7-H2 molecules are expressed on myeloma cells and, if so, whether they are associated with pathophysiology in myeloma. EXPERIMENTAL DESIGN: The expression of B7.2 and B7-H2 molecules on normal plasma and neoplastic (myeloma) plasma cells was analyzed. The cell proliferation and immunomodulatory function of myeloma cells related to B7.2 and B7-H2 expression were examined. RESULTS: Human myeloma cell lines commonly expressed B7.2 and B7-H2 molecules. B7.2 expression on plasma cells was more common in myeloma patients (n = 35) compared with that in patients with monoclonal gammopathy of unknown significance (n = 12) or hematologically normal individuals (n = 10). Plasma cells expressing B7-H2 were observed in myeloma patients alone, although rarely. Patients whose myeloma cells showed high B7.2 expression were more anemic and thrombocytopenic than other myeloma patients. The expression of these molecules was induced or augmented by cultivating myeloma cells with autologous stroma cells or tumor necrosis factor-alpha, a key cytokine in myeloma biology. Cell proliferation was more rapid in the B7.2+ and B7-H2+ populations compared with the B7.2(-) and B7 H2(-) populations, respectively, in the human myeloma cell lines examined. B7.2 and B7-H2 molecules on myeloma cells induced normal CD4+ T cells to proliferate and produce soluble factors, including interleukin-10 that stimulate myeloma cell proliferation. CONCLUSIONS: Functional B7.2 and B7-H2 molecules detected on myeloma cells may be involved in the pathophysiology of myeloma. PMID- 19188147 TI - The macrophage colony-stimulating factor 1 response signature in breast carcinoma. AB - PURPOSE: Macrophages play an important role in breast carcinogenesis. The pathways that mediate the macrophage contribution to breast cancer and the heterogeneity that exists within macrophages are incompletely understood. Macrophage colony-stimulating factor 1 (CSF1) is the primary regulator of tissue macrophages. The purpose of this study was to define a novel CSF1 response signature and to evaluate its clinical and biological significance in breast cancer. EXPERIMENTAL DESIGN: We defined the CSF1 response signature by identifying genes overexpressed in tenosynovial giant cell tumor and pigmented villonodular synovitis (tumors composed predominantly of macrophages recruited in response to the overexpression of CSF1) compared with desmoid-type fibromatosis and solitary fibrous tumor. To characterize the CSF1 response signature in breast cancer, we analyzed the expression of CSF1 response signature genes in eight published breast cancer gene expression data sets (n = 982) and did immunohistochemistry and in situ hybridization for CSF1 response genes on a breast cancer tissue microarray (n = 283). RESULTS: In both the gene microarray and tissue microarray analyses, a consistent subset (17-25%) of breast cancers shows the CSF1 response signature. The signature is associated with higher tumor grade, decreased expression of estrogen receptor, decreased expression of progesterone receptor, and increased TP53 mutations (P < 0.001). CONCLUSIONS: Our data show that the CSF1 response signature is consistently seen in a subset of breast carcinomas and correlates with biological features of the tumor. Our findings provide insight into macrophage biology and may facilitate the development of personalized therapy for patients most likely to benefit from CSF1 targeted treatments. PMID- 19188149 TI - Identification and characterization of a novel melanoma tumor suppressor gene on human chromosome 6q21. AB - PURPOSE: By characterizing a complex chromosome rearrangement involving 6q and 17p in melanoma cell line UACC-930, we isolated a candidate tumor suppressor gene at 6q21, named prenyl diphosphate synthase subunit 2 (PDSS2), which was interrupted by an inversion breakpoint. The purpose of this study was to determine the tumor-suppressive potential of PDSS2 in the development of melanoma. EXPERIMENTAL DESIGN: To isolate the rearranged 6q in UACC-930 cells, a bacterial artificial chromosome clone (RP1-67A8) covering the breakpoint at 6q21 was digested with HindIII and each DNA fragment was used as the probe for the breakpoint in Southern blotting. The HindIII fragment probe covering the breakpoint was then used to screen an EcoRI-digested DNA library generated from UACC-930. To characterize the tumor-suppressive potential of PDSS2, PDSS2 was stably transfected into a highly tumorigenic melanoma cell line, UACC-903. The tumor-suppressive function of PDSS2 was shown by both in vitro and in vivo assays. The differential expression of PDSS2 in benign nevi and primary melanoma samples was also studied. RESULTS: Down-regulation of PDSS2 was observed in 59 of 87 (67.8%) primary melanomas, which was significantly higher than that in benign nevi (7 of 66, 10.6%; P < 0.001). In addition, an overexpression of the PDSS2 in UACC-903 cells could inhibit tumor cell growth, decrease the colony-forming ability in soft agar, and totally abrogate the tumorigenicity of UACC-903 in nude mice. CONCLUSIONS: Our results support the proposal that PDSS2 is a novel tumor suppressor gene that plays an important role in the development of malignant melanoma. PMID- 19188148 TI - Nuclear versus cytoplasmic localization of filamin A in prostate cancer: immunohistochemical correlation with metastases. AB - PURPOSE: We previously showed that nuclear localization of the actin-binding protein, filamin A (FlnA), corresponded to hormone-dependence in prostate cancer. Intact FlnA (280 kDa, cytoplasmic) cleaved to a 90 kDa fragment which translocated to the nucleus in hormone-naive cells, whereas in hormone-refractory cells, FlnA was phosphorylated, preventing its cleavage and nuclear translocation. We have examined whether FlnA localization determines a propensity to metastasis in advanced androgen-independent prostate cancer. EXPERIMENTAL DESIGN: We examined, by immunohistochemistry, FlnA localization in paraffin embedded human prostate tissue representing different stages of progression. Results were correlated with in vitro studies in a cell model of prostate cancer. RESULTS: Nuclear FlnA was significantly higher in benign prostate (0.6612 +/- 0.5888), prostatic intraepithelial neoplasia (PIN; 0.6024 +/- 0.4620), and clinically localized cancers (0.69134 +/- 0.5686) compared with metastatic prostate cancers (0.3719 +/- 0.4992, P = 0.0007). Cytoplasmic FlnA increased from benign prostate (0.0833 +/- 0.2677), PIN (0.1409 +/- 0.2293), localized cancers (0.3008 +/- 0.3762, P = 0.0150), to metastases (0.7632 +/- 0.4414, P < 0.00001). Logistic regression of metastatic versus nonmetastatic tissue yielded the area under the receiver operating curve as 0.67 for nuclear-FlnA, 0.79 for cytoplasmic FlnA, and 0.82 for both, indicating that metastasis correlates with cytoplasmic to nuclear translocation. In vitro studies showed that cytoplasmic localization of FlnA induced cell invasion whereas nuclear translocation of the protein inhibited it. FlnA dephosphorylation with the protein kinase A inhibitor H-89 facilitated FlnA nuclear translocation, resulting in decreased invasiveness and AR transcriptional activity, and induced sensitivity to androgen withdrawal in hormone-refractory cells. CONCLUSIONS: The data presented in this study indicate that in prostate cancer, metastasis correlates with cytoplasmic localization of FlnA and may be prevented by cleavage and subsequent nuclear translocation of this protein. PMID- 19188150 TI - Expression of a BTB/POZ protein, NAC1, is essential for the proliferation of normal cyclic endometrial glandular cells and is up-regulated by estrogen. AB - PURPOSE: The purpose of this study was to investigate the expression and localization of NAC1, a member of the BTB/POZ gene family in the human cyclic endometrium. EXPERIMENTAL DESIGN: NAC1 expression in normal cyclic endometrium was assessed by immunohistochemistry, and data on clinical variables were collected by retrospective chart review. To elucidate the molecular mechanisms of NAC1 expression in the normal endometrium endometrial carcinoma cell lines (Ishikawa, HHUA; ER+, PR+) and primary cultured normal endometria were tested in a sex steroid induction assay and a NAC1 knockdown assay using siRNA. RESULTS: Expression of NAC1 in glandular cells was significantly higher in the early and mid proliferative phases than in the other menstrual phases. Both NAC1 RNA and protein expression were up-regulated by treatment with 10 nmol/L 17beta-Estradiol (E2) in Ishikawa, HHUA and primary cultured normal endometrial cells. The estrogen receptor antagonist ICI 182,780 significantly attenuated E2-induced NAC1 expression. NAC1 gene knockdown inhibited cell growth and induced apoptosis in Ishikawa, HHUA, and normal endometria, all of which expressed NAC1. Furthermore, NAC1 siRNA significantly abrogated estrogen-driven cellular proliferation in Ishikawa, HHUA, and primary cultured normal endometrial cells, whereas the control siRNA had no effect on cell growth in any of these cells. CONCLUSIONS: These findings suggest that NAC1 is functionally involved in E2-induced cell growth of the normal endometrial glandular cells. Because NAC1 is thought to have oncogenic potential, the current findings may provide new insight into the mechanism of estrogen induced endometrial carcinogenesis. PMID- 19188151 TI - Prognostic implications of and relationship between CpG island hypermethylation and repetitive DNA hypomethylation in hepatocellular carcinoma. AB - PURPOSE: This study aims to determine the relationship between CpG island DNA hypermethylation and global genomic DNA hypomethylation and their prognostic implications in hepatocellular carcinoma. The association of DNA methylation changes with clinicopathologic factors and the chronological ordering of DNA methylation changes along multistep hepatocarcinogenesis were also assessed. EXPERIMENTAL DESIGN: Hepatocellular carcinoma (n = 20) and nonneoplastic liver samples (n = 72) were analyzed for their methylation status at 41 CpG island loci and 3 repetitive DNA elements (LINE-1, ALU, and SAT2) using MethyLight or combined bisulfite restriction analysis. After selection of 19 CpG island loci showing cancer-specific DNA methylation, another set of 99 hepatocellular carcinoma samples was analyzed for these loci. RESULTS: The number of methylated genes in hepatocellular carcinoma was significantly higher in hepatocellular carcinoma patients with a cirrhotic liver than in hepatocellular carcinoma patients with a noncirrhotic liver (9.9 versus 7.0, P = 0.001). Hepatocellular carcinoma from female patients showed a higher number of methylated genes than hepatocellular carcinoma from male patients (11.2 versus 8.4, P = 0.006). The genes CRABP1 and SYK showed significant association between CpG island hypermethylation and patients' poor survival. SAT2 hypomethylation occurred earlier than LINE-1 or ALU hypomethylation along the multistep hepatocarcinogenesis. Depending on the type of CpG island locus, a direct, inverse, or no relationship between CpG island hypermethylation and repetitive DNA hypomethylation was observed in hepatocellular carcinomas. CONCLUSION: The varying relationships between the hypermethylation of individual CpG island loci and the hypomethylation of repetitive elements suggests that they are not mechanically linked. SYK and CRABP1 hypermethylation may serve as useful tumor markers for prognostication of hepatocellular carcinoma patients. PMID- 19188152 TI - Grape seed proanthocyanidins inhibit the growth of human non-small cell lung cancer xenografts by targeting insulin-like growth factor binding protein-3, tumor cell proliferation, and angiogenic factors. AB - PURPOSE: Lung cancer is a leading cause of cancer-related deaths worldwide. Here, we assessed the chemotherapeutic effect of grape seed proanthocyanidins (GSPs) on human non-small cell lung cancer (NSCLC) cells in vitro and in vivo using a tumor xenograft model. EXPERIMENTAL DESIGN: The effects of GSPs on human NSCLC cell lines in terms of cellular proliferation were determined. The chemotherapeutic effects of a GSP- supplemented AIN76A control diet fed to nude mice bearing tumor xenografts (A549 and H1299) were evaluated in terms of biomarkers of cell proliferation and angiogenesis and on insulin-like growth factor binding protein 3 using immunohistochemical detection, ELISA, and Western blotting. RESULTS: In vitro treatment of NSCLC cells with GSPs resulted in inhibition of cellular proliferation. Administration of GSPs (0.1%, 0.2%, and 0.5%, w/w) as a supplement of an AIN76A control diet resulted in a dose-dependent inhibition of the growth of NSCLC (A549 and H1299) tumor xenografts in athymic nude mice (25-76%; P < 0.05 0.001). The growth-inhibitory effect of GSPs on the NSCLC xenograft tumors was associated with the enhancement of the levels of insulin-like growth factor binding protein-3 in the tumor microenvironment and plasma and antiproliferative, antiangiogenic, and proapoptotic effects. CONCLUSIONS: This preclinical study reveals for the first time that dietary GSPs have the ability to inhibit the growth of human NSCLC tumor xenografts grown in vivo in athymic nude mice. More studies are needed to develop GSPs as a pharmacologically safe agent for the prevention of lung cancer in humans. PMID- 19188153 TI - CAT-8015: a second-generation pseudomonas exotoxin A-based immunotherapy targeting CD22-expressing hematologic malignancies. AB - PURPOSE: To compare the in vitro and in vivo efficacy of CAT-8015, a second generation recombinant immunotoxin composed of disulfide-linked affinity matured V(H) and V(L) chains of the mouse anti-CD22 monoclonal antibody RFB4 fused to PE38, to the parental compound CAT-3888. EXPERIMENTAL DESIGN: The biological activity of CAT-8015 was examined in vitro using B-cell tumor lines and in vivo in a JD38-based s.c. tumor model in NCr athymic mice. Pharmacokinetics and interspecies scaling of CAT-8015 were evaluated in mice, rats, and cynomolgus monkeys. The potential toxicity of CAT-8015 was assessed in monkeys in a toxicologic study and compared with CAT-3888. RESULTS: The IC50 values of CAT 8015 in vitro using the EHEB, MEC1, Daudi, CA46, and JD38 cell lines ranged from 0.3 to 8.6 ng/mL. Pharmacokinetic studies with CAT-8015 were conducted in mouse, rat, and cynomolgus monkey. The t1/2 was calculated to be 0.42, 0.61, and 0.79 hours and the Vss was 1.37, 5.57, and 140.3 mL in mouse, rat, and monkey, respectively. In vivo, when JD38 tumor-bearing animals were treated with CAT-8015 at doses > or =75 microg/kg at 48-hour intervals for a total of three doses, a rapid reduction in tumor volume and in some cases complete remission in tumor growth was observed. The comparative toxicologic study showed comparable clinical and anatomic pathology changes for CAT-8015 and CAT-3888. CONCLUSIONS: CAT-8015 is a CD22-targeting immunotoxin that, in preclinical studies, has greatly improved efficacy compared with CAT-3888. PMID- 19188155 TI - In vivo model of follicular lymphoma resistant to rituximab. AB - PURPOSE: Follicular lymphoma (FL) is the most common subtype of indolent lymphomas. Rituximab is widely used alone or in combination therapy for the treatment of FL. Despite its well-established clinical efficacy, a subpopulation of patients does not respond to rituximab and most patients will relapse after therapy. The mechanisms of action and resistance to rituximab are not fully understood. EXPERIMENTAL DESIGN: To study these mechanisms we developed an in vivo model of FL resistant to rituximab. This model was developed using the human RL line, isolated from a patient with FL, grown as xenotransplants in severe combined immunodeficient mice, exposed weekly to rituximab in vivo, followed by serial reimplantation and reexposure to rituximab, until a resistant phenotype was obtained. RESULTS: RL-derived tumors unexposed to rituximab were grown as controls and compared with the resistant tumors. Although the expression of CD46 and CD55 antigens were not differently expressed in the resistant cells, the complement inhibitor CD59 was overexpressed in a subpopulation and CD20 was found to be expressed at a lower level in a minority of cells. Bcl-X(L) and YY1 were also found more highly expressed in rituximab-resistant cells. CONCLUSION: This model provides insight on potential in vivo resistance mechanisms to rituximab and could help contribute to the development of novel therapies in rituximab refractory diseases. PMID- 19188154 TI - Dual degradation of aurora A and B kinases by the histone deacetylase inhibitor LBH589 induces G2-M arrest and apoptosis of renal cancer cells. AB - PURPOSE: This study is aimed at investigating antineoplastic efficacy of histone deacetylase inhibitor (HDACI) LBH589 on renal cell carcinoma (RCC) and elucidating the novel molecular mechanisms involved in growth arrest and apoptosis by targeting the important nonhistone molecules. EXPERIMENTAL DESIGN: We analyzed the growth-inhibitory effect of LBH589 on RCC by 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay in vitro and antitumor efficacy by xenograft experiments in vivo. To verify the associated molecular mechanisms involved in LBH589-mediated cell death and cell cycle progression by Western blotting and fluorescence-activated cell sorting analysis. RESULTS: HDACI LBH589 induced degradation of both Aurora A and B kinases through a proteasome-mediated pathway by targeting HDAC3 and HDAC6. The dual degradation of Aurora A and B kinases mediated by LBH589 resulted in inducing G2-M arrest and apoptosis of renal cancer cell lines and our results also showed that LBH589 potently inhibited renal cancer cell growth in vitro and suppressed tumor formation in vivo. The Aurora A and B kinases and HDAC3 are overexpressed in the human RCC tumor tissues examined, which make them perfect targets for HDACI LBH589 treatment. CONCLUSIONS: Our in vitro and in vivo data showed that LBH589 has potent anticancer effect of renal cancer cells. LBH589 and other HDACI treatment resulted in inducing G2-M arrest and apoptosis of renal cancer cells through degradation of Aurora A and B kinases by inhibition of HDAC3 and HDAC6. The clinical efficacy of LBH589 in the treatment of patients with metastatic RCC, especially those with high Aurora kinase and HDAC expression, is worthy of further investigation. PMID- 19188156 TI - Antitumor alkyl ether lipid edelfosine: tissue distribution and pharmacokinetic behavior in healthy and tumor-bearing immunosuppressed mice. AB - PURPOSE: The present study investigates and compares the dose-dependent pharmacokinetics and oral bioavailability of edelfosine in healthy, immunodeficient, and tumor-bearing immunosuppressed mouse animal models, as well as edelfosine uptake and apoptotic activity in the Z-138 mantle cell lymphoma (MCL) cell line. EXPERIMENTAL DESIGN: Biodistribution study of edelfosine was done in both BALB/c and severe combined immune deficiency (SCID) mice, and then the in vivo behavior of the drug after i.v. and oral administration was monitored. RESULTS: We found that edelfosine is incorporated and induces apoptosis in the Z-138 human mantle cell lymphoma cell line, whereas normal resting peripheral blood human lymphocytes were not affected. In vivo biodistribution studies revealed that accumulation of edelfosine in the tumor of a MCL-bearing mouse animal model was considerably higher (P < 0.01) than in the other organs analyzed. Besides, no statistical differences were observed between the pharmacokinetic parameters of BALB/c and SCID mice. Edelfosine presented slow elimination and high distribution to tissues. Bioavailability for a single oral dose of edelfosine was <10%, but a multiple-dose oral administration increased this value up to 64%. CONCLUSION: Our results show that edelfosine is widely scattered across different organs, but it is preferentially internalized by the tumor both in vitro and in vivo. Our data, together with the apoptotic action of the drug on cancer cells, support a rather selective action of edelfosine in cancer treatment, and that multiple oral administration is required to increase oral bioavailability. PMID- 19188157 TI - Inhibition of micrometastatic prostate cancer cell spread in animal models by 213Bilabeled multiple targeted alpha radioimmunoconjugates. AB - PURPOSE: To investigate the therapeutic potential of 213Bilabeled multiple targeted alpha-radioimmunoconjugates for treating prostate cancer (CaP) micrometastases in mouse models. EXPERIMENTAL DESIGN: PC-3 CaP cells were implanted s.c., in the prostate, and intratibially in NODSCID mice. The expression of multiple tumor-associated antigens on tumor xenografts and micrometastases was detected by immunohistochemistry. Targeting vectors were two monoclonal antibodies, and a plasminogen activator inhibitor type 2 that binds to cell surface urokinase plasminogen activator, labeled with 213Bi using standard methodology. In vivo efficacy of multiple alpha conjugates (MTAT) at different activities was evaluated in these mouse models. Tumor growth was monitored during observations and local regional lymph node metastases were assessed at the end of experiments. RESULTS: The take rate of PC-3 cells was 100% for each route of injection. The tumor-associated antigens (MUC1, urokinase plasminogen activator, and BLCA-38) were heterogeneously expressed on primary tumors and metastatic cancer clusters at transit. A single i.p. injection of MTAT (test) at high and low doses caused regression of the growth of primary tumors and prevented local lymph node metastases in a concentration-dependent fashion; it also caused cancer cells to undergo necrosis and apoptosis. CONCLUSIONS: Our results suggest that MTAT can impede primary PC-3 CaP growth at three different sites in vivo through induction of apoptosis, and can prevent the spread of cancer cells and target lymph node micrometastases in a concentration-dependent manner. MTAT, by targeting multiple antigens, can overcome heterogeneous antigen expression to kill small CaP cell clusters, thus providing a potent therapy for micrometastases. PMID- 19188158 TI - Targeted photothermal ablation of murine melanomas with melanocyte-stimulating hormone analog-conjugated hollow gold nanospheres. AB - PURPOSE: To develop melanoma-targeted hollow gold nanospheres (HAuNS) and evaluate their potential utility for selective photothermal ablation in melanoma. EXPERIMENTAL DESIGN: A new class of photothermal coupling agents based on HAuNS was synthesized. HAuNS were stabilized with polyethylene glycol (PEG) coating and attached with alpha-melanocyte-stimulating hormone (MSH) analog, [Nle4,D Phe7]alpha-MSH (NDP-MSH), which is a potent agonist of melanocortin type-1 receptor overexpressed in melanoma. The intracellular uptake of the NDP-MSH conjugated PEGylated HAuNS (NDP-MSH-PEG-HAuNS) and the distribution of beta arrestin were examined in murine B16/F10 melanoma cells. The biodistribution of NDP-MSH-PEG-HAuNS was assessed at 4 hours post i.v. injection in tumor-bearing nude mice. Photothermal ablation effect of the nanoparticles was evaluated both histologically using excised tissue and functionally by [18F]fluorodeoxyglucose positron emission tomography. RESULTS: NDP-MSH-PEG-HAuNS consist only of a thin gold wall with hollow interior (outer diameter, 43.5 +/- 2.3 nm; shell thickness, 3-4 nm), which displays strong and tunable resonance absorption in near-IR region (peak, 808 nm). The nanoparticles were specifically taken up by melanoma cells, which initiated the recruitment of beta-arrestins, the adapters to link the activated G-protein-coupled receptors to clathrin, indicating the involvement of receptor-mediated endocytosis. This resulted in enhanced extravasation of NDP-MSH PEG-HAuNS from tumor blood vessels and their dispersion into tumor matrix compared with nonspecific PEGylated HAuNS. Successful selective photothermal ablation of B16/F10 melanoma with targeted HAuNS was confirmed by histologic and [18F]fluorodeoxyglucose positron emission tomography evaluation at 24 hours post near IR-region laser irradiation at a low-dose energy of 30 J/cm2. CONCLUSION: NDP-MSH-PEG-HAuNS have the potentials to mediate targeted photothermal ablation of melanoma. PMID- 19188159 TI - Potential therapeutic effect of glycogen synthase kinase 3beta inhibition against human glioblastoma. AB - PURPOSE: Glioblastoma represents the malignant brain tumor that is most refractory to treatment and in which the identification of molecular target(s) is urgently required. We investigated the expression, activity, and putative pathologic role of glycogen synthase kinase 3beta (GSK3beta), an emerging therapeutic target for neurodegenerative diseases, in human glioblastoma. EXPERIMENTAL DESIGN: The active fraction of GSK3beta that is phosphorylated at the tyrosine 216 residue (pGSK3betaY216) was identified in glioblastoma cell lines. GSK3beta activity for phosphorylating its substrate was detected in these cells by nonradioisotopic in vitro kinase assay. RESULTS: Higher expression levels of GSK3beta and pGSK3betaY216 were frequently detected in glioblastomas compared with nonneoplastic brain tissues. Inhibition of GSK3beta activity by escalating doses of a small-molecule inhibitor (AR-A014418) or inhibition of its expression by RNA interference induced the apoptosis and attenuated the survival and proliferation of glioblastoma cells in vitro. Inhibition of GSK3beta was associated with increased expression of p53 and p21 in glioblastoma cells with wild-type p53 and with decreased Rb phosphorylation and expression of cyclin dependent kinase 6 in all glioblastoma cell lines. Administration of AR-A014418 at a low dose significantly sensitized glioblastoma cells to temozolomide and 1 (4-amino-2-methyl-5-pyrimidinyl)methyl-3-(2-chloroethyl)-3-nitrosourea, chemotherapeutic agents used in the clinical setting, as well as to ionizing radiation. CONCLUSION: These results indicate that GSK3beta exerts a pathologic role by promoting the survival and proliferation of glioblastoma cells and by protecting them from apoptosis via the inactivation of p53- and/or Rb-mediated pathways. Consequently, we propose that GSK3beta provides a potential therapeutic target in glioblastoma. PMID- 19188160 TI - In vitro and in vivo anticancer effects of the novel vitamin E ether analogue RRR alpha-tocopheryloxybutyl sulfonic acid in prostate cancer. AB - PURPOSE: Among derivatives of alpha-vitamin E, alpha-vitamin E succinate (VES), has attracted much attention due to its potent anti-prostate cancer activity in vitro and in vivo. However, the in vivo antitumor activity of VES might be compromised if administrated orally due to the VES hydrolysis by esterases in the gastrointestinal tract. EXPERIMENTAL DESIGN: New nonhydrolyzable VES ether analogues were synthesized and their growth inhibition was screened by 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide growth assay. Among them, RRR-alpha-tocopheryloxybutyl sulfonic acid (VEBSA) was further characterized by terminal deoxyribonucleotidyl transferase-mediated dUTP nick end labeling apoptosis assay, soft agar assay, and in vivo tumor formation. RESULTS: VEBSA has potent antitumor ability, albeit to a lesser extent than VES, in in vitro cultured prostate cancer LNCaP and PC3 cells. Like VES, VEBSA induced apoptosis, repressed androgen receptor protein expression, and enhanced vitamin D receptor expression, suggesting that VEBSA can go through mechanisms similar to those used by VES to inhibit the growth of prostate cancer cells in vitro. However, 6 weeks of oral consumption of VEBSA, but not of VES, reduced the tumor burden in the xenografted prostate tumors in nude mice. Furthermore, oral intake of VEBSA for 20 weeks inhibited prostate tumor growth and progression more efficiently compared with VES in the prostate cancer tumor model of TRAMP mice. CONCLUSION: Oral consumption of VEBSA allows a greater anticancer activity compared with VES. Chemoprevention prefers the oral consumption of agents; the advantage of VEBSA over VES to be administrated orally will allow VEBSA to serve as an agent for both preventive and therapeutic purposes for prostate cancer. PMID- 19188161 TI - Addition of S-1 to the epidermal growth factor receptor inhibitor gefitinib overcomes gefitinib resistance in non-small cell lung cancer cell lines with MET amplification. AB - PURPOSE: Most non-small cell lung cancer (NSCLC) tumors with activating mutations in the epidermal growth factor receptor (EGFR) are initially responsive to EGFR tyrosine kinase inhibitors (EGFR-TKI) such as gefitinib and erlotinib, but they almost invariably develop resistance to these drugs. A secondary mutation in EGFR (T790M) and amplification of the MET proto-oncogene have been identified as mechanisms of such acquired resistance to EGFR-TKIs. We have now investigated whether addition of the oral fluoropyrimidine derivative S-1 to gefitinib might overcome gefitinib resistance in NSCLC cell lines. EXPERIMENTAL DESIGN: The effects of gefitinib on EGFR signaling and on the expression both of thymidylate synthase and of the transcription factor E2F-1 in gefitinib-resistant NSCLC cells were examined by immunoblot analysis. The effects of S-1 (or 5-fluorouracil) and gefitinib on the growth of NSCLC cells were examined in vitro as well as in nude mice. RESULTS: Gefitinib induced down-regulation of thymidylate synthase and E2F 1 in gefitinib-resistant NSCLC cells with MET amplification but not in those harboring the T790M mutation of EGFR. The combination of 5-fluorouracil and gefitinib synergistically inhibited the proliferation of cells with MET amplification, but not that of those with the T790M mutation of EGFR, in vitro. Similarly, the combination of S-1 and gefitinib synergistically inhibited the growth only of NSCLC xenografts with MET amplification. CONCLUSIONS: Our results suggest that the addition of S-1 to EGFR-TKIs is a promising strategy to overcome EGFR-TKI resistance in NSCLC with MET amplification. PMID- 19188162 TI - Ionizing radiation up-regulates telomerase activity in cancer cell lines by post translational mechanism via ras/phosphatidylinositol 3-kinase/Akt pathway. AB - PURPOSE: Telomerase is considered currently as a hallmark of cancer, and its inhibition is expected to become an important anticancer modality. In contrast to abundant data concerning the effect of cytotoxic drugs on telomerase activity (TA), there is scant information on the effect of radiation on telomerase. The mechanism of telomerase regulation by irradiation has never been evaluated in detail. In the present study, we investigated the effect of radiation on TA and its regulation in cancer cells. EXPERIMENTAL DESIGN: The effect of various radiation doses on TA in several malignant and nonmalignant cell lines was evaluated. All malignant cells exhibited similar telomerase response to radiation and its regulation was assessed at transcriptional and post-translational levels in K562 cells. Next step was the evaluation of the upstream signaling pathways leading to changes in TA using kinetics and specific inhibitors. RESULTS: Radiation up-regulated TA in dose-dependent manner only in cancer cells. Telomerase was activated by phosphorylation by Akt and by cytoplasmic-nuclear shift. Transcriptional processes were not involved in TA. This telomerase regulation is mediated by Ras/phosphatidylinositol 3-kinase/Akt pathway. The canonical membrane effectors of irradiation (epidermal growth factor receptor, insulin-like growth factor-I receptor, and Ca2+ influx) were not involved in this process. CONCLUSIONS: Radiation up-regulates telomerase activity specifically in cancer cells. This study adds to accumulating evidence pointing to post translational level as important mode of telomerase regulation. Telomerase activation due to radiation may be detrimental in treatment of cancer. Data described in this study may add to future interventions aiming at inhibition of telomerase activation during irradiation. PMID- 19188163 TI - A novel human Her-2/neu chimeric molecule expressed by Listeria monocytogenes can elicit potent HLA-A2 restricted CD8-positive T cell responses and impact the growth and spread of Her-2/neu-positive breast tumors. AB - PURPOSE: The aim of this study was to efficiently design a novel vaccine for human Her-2/neu-positive (hHer-2/neu) breast cancer using the live, attenuated bacterial vector Listeria monocytogenes. EXPERIMENTAL DESIGN: Three recombinant L. monocytogenes-based vaccines were generated that could express and secrete extracellular and intracellular fragments of the hHer-2/neu protein. In addition, we generated a fourth construct fusing selected portions of each individual fragment that contained most of the human leukocyte antigen (HLA) epitopes as a combination vaccine (L. monocytogenes-hHer-2/neu chimera). RESULTS: Each individual vaccine was able to either fully regress or slow tumor growth in a mouse model for Her-2/neu-positive tumors. All three vaccines could elicit immune responses directed toward human leukocyte antigen-A2 epitopes of hHer-2/neu. The L. monocytogenes-hHer-2/neu chimera was able to mimic responses generated by the three separate vaccines and prevent spontaneous outgrowth of tumors in an autochthonous model for Her-2/neu-positive breast cancer, induce tumor regression in transplantable models, and prevent seeding of experimental lung metastases in a murine model for metastatic breast cancer. CONCLUSION: This novel L. monocytogenes-hHer-2/neu chimera vaccine proves to be just as effective as the individual vaccines but combines the strength of all three in a single vaccination. These encouraging results support future clinical trials using this chimera vaccine and may be applicable to other cancer types expressing the Her 2/neu molecule such as colorectal and pancreatic cancer. PMID- 19188164 TI - MDM2 antagonist nutlin-3 displays antiproliferative and proapoptotic activity in mantle cell lymphoma. AB - PURPOSE: Mantle cell lymphoma (MCL) has one of the poorest prognoses of the non Hodgkin's lymphomas, and novel therapeutic approaches are needed. We wished to determine whether Nutlin-3, a novel small-molecule murine double minute 2 (MDM2) antagonist that efficiently activates TP53, might be effective in inducing cell death in MCL. EXPERIMENTAL DESIGN: MCL cell lines with known TP53 status were treated with Nutlin-3, and biological and biochemical consequences were studied. Synergies with the prototypic genotoxic agent doxorubicin and the novel proteasome inhibitor bortezomib were assessed. RESULTS: Nutlin-3 resulted in a reduction in cell proliferation/viability (IC50 < 10 micromol/L), an increase in the apoptotic fraction, and cell cycle arrest in wild-type (wt) TP53 Z-138 and Granta 519 cells. These effects were accompanied by TP53 accumulation and induction of TP53-dependent proteins p21, MDM2, Puma, and Noxa. Cell cycle arrest was characterized by suppression of S phase and an increase in the G0-G1 and G2-M fractions and accompanied by suppression of total and phosphorylated retinoblastoma protein and a decrease in G2-M-associated proteins cyclin B and CDC2. The combination of Nutlin-3 with doxorubicin or bortezomib was synergistic in wt-TP53 MCL cells. Nutlin-3 also induced cell cycle arrest and reduced cell viability in the mutant TP53 MINO cells but at a significantly higher IC50 (22.5 micromol/L). These effects were associated with induction of the TP53 homologue p73, slight increases in p21 and Noxa, and caspase activation. Nutlin-3 and bortezomib synergistically inhibited cell growth of MINO. CONCLUSION: These findings suggest that the MDM2 antagonist Nutlin-3 may be an effective agent in the treatment of MCL with or without wt-TP53. PMID- 19188165 TI - Genetically targeted T cells eradicate established breast cancer in syngeneic mice. AB - PURPOSE: The purpose of the present study was to evaluate the capacity and mechanisms of genetically modified erbB2-specific T cells to eradicate erbB2+ tumors in syngeneic mice. EXPERIMENTAL DESIGN: Primary mouse T cells were modified to target the breast tumor-associated antigen erbB2 through retroviral mediated transfer of a chimeric antigen receptor, termed single-chain antibody (scFv)-CD28-zeta. Antitumor efficacy of scFv-CD28-zeta-modified T cells was analyzed in mice bearing D2F2/E2 breast tumors. RESULTS: The scFv-CD28-zeta modified T cells were shown to specifically secrete T cytotoxic-1 cytokines and lyse erbB2+ breast tumor cells following receptor stimulation in vitro. Treatment with scFv-CD28-zeta-modified T cells was able to lead to long-term, tumor-free survival in mice bearing erbB2+ D2F2/E2 breast tumors. Importantly, the surviving mice developed a host memory response to D2F2/E2 tumor cells, and this host response was able to protect against a rechallenge with erbB2+ D2F2/E2 tumor cells and parental erbB2(-) D2F2 tumor cells. In addition, scFv-CD28-zeta T-cell expression of perforin and interferon-gamma were essential for complete antitumor efficacy. CONCLUSIONS: Treatment with scFv-CD28-zeta-modified T cells was able to induce a host antitumor immunity in syngeneic mice. Complete tumor elimination by scFv-CD28-zeta-modified T cells required T cell-derived interferon-gamma and perforin, indicating that cytotoxicity and cytokine secretion play a role in the in vivo response. PMID- 19188166 TI - Human-like mouse models for testing the efficacy and safety of anti-beta2 microglobulin monoclonal antibodies to treat myeloma. AB - PURPOSE: We showed recently that anti-beta2-microglobulin (beta2M) monoclonal antibodies (mAb) have remarkably strong apoptotic effects on myeloma cells in vitro and in SCID-hu mice. However, whether the mAbs will be therapeutic and safe in the treatment of myeloma patients, in whom every tissue expresses low densities of MHC class I molecules and elevated levels of soluble beta2M are present, remains to be determined. EXPERIMENTAL DESIGN: In this study, human-like myeloma mouse models (HLA-A2-transgenic NOD/SCID mice) were developed, which express mature and functional human MHC class I (HLA-A2 and human beta2M) on murine organs and present high levels of circulating human beta2M derived from human myeloma cells. Myeloma-bearing mice were treated intraperitoneally with anti-beta2M mAbs, and the distribution and effects of the mAbs on normal organs and established tumors were examined. RESULTS: Our results show that anti-beta2M mAbs were effective in suppressing myeloma growth in treated mice. The therapeutic efficacy of the mAbs in these mice are comparable with those observed in myeloma-bearing nontransgenic NOD/SCID mice in which no human MHC class I is expressed on murine organs. Furthermore, although the mAbs can be detected on different organs, no tissue damage or cell apoptosis was observed in the mice. CONCLUSION: Based on the antimyeloma efficacy and low toxicity in the mice, our study suggests that anti-beta2M mAbs may be safe and the tissue-expressing and soluble beta2M may not compromise their therapeutic effects in myeloma patients. This study provides further support for the future application of the mAbs as therapeutic agents for multiple myeloma. PMID- 19188167 TI - B7 blockade alters the balance between regulatory T cells and tumor-reactive T cells for immunotherapy of cancer. AB - PURPOSE: In prostate cancer-bearing host, regulatory T (Treg) cells restrain activity of tumor antigen-specific T cells. Because B7:CD28 interactions are needed for both function of CD4+CD25+ Treg cells and CD8+ effective T cells, targeting this pathway may help to overcome the immunotherapy barriers. EXPERIMENTAL DESIGN: The anti-B7-1/B7-2 monoclonal antibodies were administered to a transgenic mouse model of prostate cancer (TRAMP) ectopically expressing SV40 large T antigen in different tumor development stages for prevention and therapy of prostate cancer. The treatment was also tested in treating transplanted MC38 colon adenocarcinoma in mice. RESULTS: Here, we showed that short-term administration of anti-B7-1/B7-2 monoclonal antibodies in TRAMP mice leads to significant inhibited primary tumor growth and the size of metastatic lesions. The treatment is effective to inhibit MC38 colon cancer growth. Correspondingly, this treatment results in a transient reduction of Treg in both thymus and the periphery. In vivo cytotoxicity assay revealed T antigen-specific CTL effectors in anti-B7-treated but not control IgG-treated TRAMP mice. CONCLUSIONS: Transient blockade of B7-1/B7-2 alters the balance between Treg and cancer-reactive T cells to enhance cancer immunotherapy. PMID- 19188168 TI - Overexpression of PD-L1 significantly associates with tumor aggressiveness and postoperative recurrence in human hepatocellular carcinoma. AB - PURPOSE: The aberrant expression of programmed cell death 1 ligands 1 and 2 (PD Ls) on tumor cells dampens antitumor immunity, resulting in tumor immune evasion. In this study, we investigated the expression of PD-Ls in human hepatocellular carcinoma (HCC) to define their prognostic significance after curative surgery. EXPERIMENTAL DESIGN: Immunohistochemistry was used to investigate PD-Ls expression as well as granzyme B+ cytotoxic and FoxP3+ regulatory T cell infiltration on tissue microarrays containing 240 randomly selected HCC patients who underwent surgery. The results were further verified in an independent cohort of 125 HCC patients. PD-Ls expression on HCC cell lines was detected by Western blot assay. RESULTS: Patients with higher expression of PD-L1 had a significantly poorer prognosis than patients with lower expression. Although patients with higher expression of PD-L2 also had a poorer survival, the difference in recurrence was not statistically significant. Multivariate analysis identified tumor expression of PD-L1 as an independent predictor for postoperative recurrence. No correlation was found between PD-Ls expression and granzyme B+ lymphocyte infiltration, whereas a significant positive correlation was detected between PD-Ls expression and FoxP3+ lymphocyte infiltration. In addition, tumor infiltrating cytotoxic and regulatory T cells were also independent prognosticators for both survival and recurrence. The prognostic value of PD-L1 expression was validated in the independent data set. CONCLUSION: Our data suggest for the first time that PD-L1 status may be a new predictor of recurrence for HCC patients and provide the rationale for developing a novel therapy of targeting the PD-L1/PD-1 pathway against this fatal malignancy. PMID- 19188169 TI - Differential diagnosis of pulmonary carcinoma following head and neck cancer by genetic analysis. AB - PURPOSE: Patients with head and neck cancer often develop a lung tumor that can be diagnosed as distant metastasis (DM) or second primary tumor (SPT). In this study, we use TP53 mutation analysis for validation of an allelic loss marker panel and a decision algorithm for distinguishing between DM and SPT. EXPERIMENTAL DESIGN: Tumor pairs of 39 patients were analyzed for TP53 mutations, for patterns of allelic loss and immunohistochemical analysis of p53 expression. Results of these three analyses were compared, using mutation analysis as gold standard. RESULTS: Loss of heterozygosity (LOH) analysis indicated DM in 15 and SPT in 23 cases (one inconclusive). TP53 mutation analysis was informative in 15 cases. Based on the p53 mutation status alone, nine tumors were diagnosed as SPT and six as DM. In all 15 cases the LOH analysis was in concordance with the TP53 mutation analysis. Immunostaining for p53 showed promise as a first scan to diagnose lung tumors as SPT but cannot be used to diagnose DM. CONCLUSION: The TP53 mutation data validate the suitability of the LOH marker panel and decision algorithm for differential diagnosis of DM and SPT in the lung. LOH analysis can theoretically be exploited in almost all cases and is less laborious than TP53 mutation analysis. PMID- 19188170 TI - Diffusion-weighted magnetic resonance imaging for predicting and detecting early response to chemoradiation therapy of squamous cell carcinomas of the head and neck. AB - PURPOSE: The aim of this study was to investigate the utility of apparent diffusion coefficient (ADC) for prediction and early detection of treatment response in head and neck squamous cell carcinomas (HNSCC). EXPERIMENTAL DESIGN: Diffusion-weighted magnetic resonance imaging studies were performed on 40 patients with newly diagnosed HNSCC before, during, and after the end of chemoradiation therapy. Analysis was done on data from 33 patients after exclusion of 7 patients that had incomplete data. RESULTS: Pretreatment ADC value of complete responders (1.04 +/- 0.19 x 10(-3) mm2/s) was significantly lower (P < 0.05) than that from partial responders (1.35 +/- 0.30 x 10(-3) mm2/s). A significant increase in ADC was observed in complete responders within 1 week of treatment (P < 0.01), which remained high until the end of the treatment. The complete responders also showed significantly higher increase in ADC than the partial responders by the first week of chemoradiation (P < 0.01). When pretreatment ADC value was used for predicting treatment response, the area under the receiver operating characteristic curve was 0.80 with a sensitivity of 65% and a specificity of 86%. However, change in ADC within the first week of chemoradiation therapy resulted in an area under the receiver operating characteristic curve of 0.88 with 86% sensitivity and 83% specificity for prediction of treatment response. CONCLUSIONS: These results suggest that ADC can be used as a marker for prediction and early detection of response to concurrent chemoradiation therapy in HNSCC. PMID- 19188171 TI - The prognostic value of TP53 mutations in chronic lymphocytic leukemia is independent of Del17p13: implications for overall survival and chemorefractoriness. AB - PURPOSE: Del17p13 predicts poor outcome and chemorefractoriness in chronic lymphocytic leukemia (CLL). Conversely, it is unknown whether TP53 mutations carry any prognostic value independent of del17p13. We tested the independent prognostic value of TP53 mutations in CLL. EXPERIMENTAL DESIGN: The study was based on a consecutive series of 308 CLL. DNA sequencing of TP53 exons 2 to 10 and del17p13 interphase fluorescence in situ hybridization were done at CLL diagnosis. Study end points were survival and chemorefractoriness. RESULTS: At diagnosis, TP53 mutations (n = 32) occurred in 31 of 308 (10.0%) patients. Of all CLL showing TP53 disruption by either mutation and/or deletion (n = 44), 10 cases (22.7%) showed TP53 mutations in the absence of del17p13. Multivariate analysis selected TP53 mutations (hazard ratio, 3.20; P = 0.002) as an independent predictor of overall survival after adjustment for del17p13. Also, multivariate analysis selected TP53 mutations (hazard ratio, 3.97; P < 0.001) as an independent predictor of chemorefractoriness after adjustment for del17p13. Compared with cases without TP53 alterations, CLL harboring any type of TP53 disruption (mutation only, del17p13 only, or both mutation and del17p13) uniformly displayed a high prevalence of unfavorable prognosticators and poor outcome. Analysis of sequential CLL samples showed the acquisition of new or additional TP53 alterations at the time of chemorefractoriness. CONCLUSIONS: These data show that (a) TP53 mutations are an independent predictor of short survival and chemorefractoriness, and (b) that CLL presenting with TP53 mutations without del17p13 fare as poorly as CLL carrying del17p13. Because CLL harboring TP53 mutations without del17p13 are currently not recognized by conventional diagnostic strategies, these results may be relevant for a comprehensive prognostic characterization of CLL. PMID- 19188172 TI - Clinical significance of cytokeratin 20-positive circulating tumor cells detected by a refined immunomagnetic enrichment assay in colorectal cancer patients. AB - PURPOSE: Current immunomagnetic enrichment method can only detect general epithelial antigens of circulating tumor cells (CTC). Further characterization of the CTCs to provide specific information on the tumor type is not possible. We attempted to overcome this drawback by developing the methodology for using a gastrointestinal-specific anti-cytokeratin (CK) 20 antibody to detect CTCs in colorectal cancer patients' blood. EXPERIMENTAL DESIGN: The protocol was validated using a colorectal cancer SW480 cell line. The clinical significance of findings in colorectal cancer was investigated by detecting CK20-positive CTCs (pCTC) in patients with colorectal cancer, other common cancers, colorectal adenoma, benign colorectal diseases, and normal subjects. Moreover, the malignant nature of CK20 pCTCs was examined by comparing chromosome 17 aberration patterns with those from the corresponding primary tumors. RESULTS: The assay successfully showed CK20-positive SW480 cells. When applied in patient samples, the detection rates were 62% (132 colorectal cancer patients; median number = 11 CTCs), 0% (120 patients with other common cancers), 6% (50 colorectal adenoma patients), 0% (120 patients with benign colorectal diseases), and 0% (40 normal subjects). Furthermore, statistical analysis showed that CK20 pCTC numbers were associated with tumor-node-metastasis stage and lymph node status. Using the median CK20 pCTC numbers as the cutoff points, stratified groups of colorectal cancer patients had significant differences in their recurrence, metastasis, and survival. Finally, chromosome 17 aneusomy in 90% of colorectal cancer patients with CK20 pCTCs matched with those from the primary tumors. CONCLUSIONS: Detection of CK20 pCTCs using the new protocol could generate clinically important information for colorectal cancer patients. PMID- 19188173 TI - A simple and accurate model for prediction of cancer-specific mortality in patients treated with surgery for primary penile squamous cell carcinoma. AB - PURPOSE: Cancer-specific mortality (CSM) of patients with primary penile squamous cell carcinoma (PPSCC) may be quite variable. Recently, a nomogram was developed to provide standardized and individualized mortality predictions. Unfortunately, it relies on a large number (n = 8) of specific variables that are unavailable in routine clinical practice. We attempted to develop a simpler prediction rule with at least equal accuracy in predicting CSM after surgical removal of PPSCC. EXPERIMENTAL DESIGN: The predictive rule was developed on a cohort of 856 patients identified in the 1988 to 2004 Surveillance, Epidemiology and End Results (SEER) database. The predictors consisted of age, race, SEER stage (localized versus regional versus metastatic), tumor grade, type of surgery (excisional biopsy, partial penectomy, and radical penectomy), and of lymph node status (pN0 versus pN1-3 versus pNx). A look-up table based on Cox regression model-derived coefficients was used for prediction of 5-year CSM. The predictive rule accuracy was tested using the Harrell's modification of the area under the receiver operating characteristics curve. RESULTS: SEER stage and histologic grade achieved independent predictor status and qualified for inclusion in the model. The model achieved 73.8% accuracy for prediction of CSM at 5 years after surgery. Both predictors achieved independent predictor status in competing risk regression models addressing CSM, where other cause mortality was controlled for. CONCLUSION: Despite equivalent accuracy, our predictive rule predicting 5-year CSM in patients with PPSCC is substantially less complex (2 versus 8 variables) than the previously published model. PMID- 19188174 TI - Genetic variants in TAP are associated with high-grade cervical neoplasia. AB - PURPOSE: The transporter associated with antigen processing (TAP) is essential in assembling MHC-I proteins. Human papillomavirus (HPV) evades immune recognition by decreasing class I MHC cell surface expression through down-regulation of TAP1 levels. Consistent with heterogeneity in MHC expression is the individual variability in clearing detectable HPV infections. Genetic polymorphisms in TAP genes may affect protein structure, function, and the ability to clear HPV infection. EXPERIMENTAL DESIGN: Case-control study of women with cervical intraepithelial neoplasia (CIN) II or III (n = 114) and women without high-grade CIN (n = 366). Five nonsynonymous single nucleotide polymorphisms (SNP) in TAP1 and TAP2 were genotyped using DNA collected in cervicovaginal lavage samples using microsphere array technology (Luminex xMAP). HPV typing was done using a PCR-based system with MY09/MY11 primers. TAP1 and TAP2 SNPs were validated by direct sequencing. RESULTS: Differences in allele distribution between women with high-grade cervical neoplasia and women without was seen for TAP1 I333V (P = 0.02) and TAP1 D637G (P = 0.01). The odds ratios (OR) for CIN III were significantly lower among carriers of the TAP1 I333V polymorphism (OR, 0.28; 95% confidence interval, 0.1-0.8), and TAP1 D637G polymorphism (OR, 0.27; 95% confidence interval, 0.1-0.7). These associations remained significant even after restricting the evaluation to women who were positive for high-risk HPV types. CONCLUSIONS: In addition to the down-regulation of MHC-1 by oncogenic HPV, HPV pathogenesis might be facilitated by polymorphisms in the TAP proteins. Identifying TAP polymorphisms may potentially be used to identify women less susceptible to progression to high-grade CIN and cervical cancer. PMID- 19188176 TI - Cell-free tumor DNA in blood plasma as a marker for circulating tumor cells in prostate cancer. AB - PURPOSE: Circulating cell-free DNA in the blood of cancer patients harbors tumor specific aberrations. Here, we investigated whether this DNA might also reflect the presence of circulating tumor cells (CTC). EXPERIMENTAL DESIGN: To identify the source of cell-free DNA in blood, plasma derived from 81 patients with prostate cancer was examined for CTCs and cell-free DNA. An epithelial immunospot assay was applied for detection of CTCs, and a PCR-based fluorescence microsatellite analysis with a panel of 14 polymorphic markers was used for detection of allelic imbalances (AI). RESULTS: The plasma DNA levels significantly correlated with the diagnosis subgroups of localized (stage M0, n = 69) and metastasized prostate cancer (stage M1, n = 12; P = 0.03) and with the tumor stage of these patients (P < 0.005). AI was found on cell-free DNA in plasma from 45.0% and 58.5% of M0 and M1 patients, respectively. Detection of CTCs showed that 71.0% or 92.0% of the M0 and M1 patients harbored 1 to 40 CTCs in their blood, respectively. The occurrence of CTCs correlated with tumor stage (P < 0.03) and increasing Gleason scores (P = 0.04). Notably, significant associations of the number of CTCs with the AI frequencies at the markers D8S137 (P = 0.03), D9S171 (P = 0.04), and D17S855 (P = 0.02) encoding the cytoskeletal protein dematin, the inhibitor of the cyclin-dependent kinase CDKN2/p16 and BRCA1, respectively, were observed. CONCLUSIONS: These findings show, for the first time, a relationship between the occurrence of CTCs and circulating tumor associated DNA in blood, which, therefore, might become a valuable new source for monitoring metastatic progression in cancer patients. PMID- 19188175 TI - Cystatin B as a tissue and urinary biomarker of bladder cancer recurrence and disease progression. AB - PURPOSE: Using proteomic techniques, we sought to identify novel protein biomarkers in tissue and urine from patients with transitional cell carcinoma (TCC). EXPERIMENTAL DESIGN: Urinary and tissue proteomes were analyzed and differentially expressed proteins were identified by mass spectrometry. One of the proteins, cystatin B, was further analyzed in TCC tissue by immunohistochemistry and in urine by semiquantitative Western blot analysis. RESULTS: Cystatin B tissue staining intensity significantly increased concordantly with TCC grade (P = 0.0008). Elevated urinary cystatin B levels correlated with increasing tumor grade (P = 0.062) and stage (P = 0.0047). Patients with elevated levels of cystatin B had a shorter mean +/- SE time to disease recurrence (12 +/- 1.82 months) compared with patients who had low levels (28.8 +/- 2.26 months; P = 0.0047). Similarly, patients with elevated cystatin B levels had a shorter time to grade/stage progression compared with patients with low urinary cystatin B (P = 0.0007). By multivariate Cox regression analysis, an elevated cystatin B level was the most significant variable predicting disease recurrence (hazard ratio, 3.8; 95% confidence interval, 1.5-9.5; P = 0.0049) and grade/stage progression (hazard ratio, 10.4; 95% confidence interval, 1.6-201.5; P = 0.0104). CONCLUSIONS: Cystatin B is elevated in tissue and urine of bladder cancer patients. Cystatin B urine levels are positively correlated with tumor grade, stage, and shorter time to disease recurrence and progression. Consequently, cystatin B may be useful as a novel predictive biomarker in TCC of the bladder. PMID- 19188177 TI - Polymerase eta mRNA expression predicts survival of non-small cell lung cancer patients treated with platinum-based chemotherapy. AB - PURPOSE: The effect of translesion DNA synthesis system in conferring cellular tolerance to DNA-damaging agents has been recently described. DNA polymerase eta (Pol eta) is part of this machinery and in vitro models showed that it can overcome DNA damages caused by cisplatin and UV rays. The aim of the present study was to investigate the role of Pol eta mRNA expression levels in non-small cell lung cancer (NSCLC). EXPERIMENTAL DESIGN: Pol eta mRNA expression levels were evaluated by real-time PCR in (a) formalin-fixed paraffin-embedded biopsies of 72 NSCLC patients treated with platinum-based chemotherapy, (b) fresh snap frozen surgical specimens of tumor and corresponding normal lung tissue from 50 consecutive patients not treated with perioperative or postoperative chemotherapy, and (c) five NSCLC cell lines. RESULTS: High Pol eta expression levels were strongly associated with shorter survival at both univariate (6.9 versus 21.1 months; P = 0.003) and multivariate (hazard ratio, 3.18; 95% confidence interval, 1.73-5.84; P = 0.008) analysis in the group of platinum treated patients. By contrast, Pol eta expression was not significantly correlated with the prognosis in surgically resected patients (P = 0.54) and mRNA levels did not significantly differ in tumor versus normal lung (P = 0.82). Moreover, endogenous Pol eta mRNA expression was found to be inducible by cisplatin in three of five cell lines and significantly associated with in vitro sensitivity (P = 0.01). CONCLUSIONS: Taken together, these data indicate Pol eta as a predictive rather than prognostic marker worth of further investigation in NSCLC patients candidate to platinum-based chemotherapy. PMID- 19188178 TI - Immunomodulation of FOXP3+ regulatory T cells by the aromatase inhibitor letrozole in breast cancer patients. AB - PURPOSE: We have shown previously that tumor infiltration by FOXP3+ regulatory T cells (Treg) is associated with increased relapse and shorter survival of patients with both in situ and invasive breast cancer. Because estrogen regulates Treg numbers in mice and promotes the proliferation of human Tregs, we hypothesized that blocking estrogen receptor-alpha signaling would abrogate Tregs and be associated with response to hormonal therapy and increased survival. EXPERIMENTAL DESIGN: FOXP3+ Tregs were quantified in tumor samples collected at baseline by incisional biopsy and after 6 months at definitive surgery in 83 elderly breast cancer patients (T2-4 N0-1) enrolled in a randomized phase II trial based on 6 months of primary letrozole (2.5 mg/d) or 6 months of letrozole plus oral "metronomic" cyclophosphamide (50 mg/d). RESULTS: Treg number ranged from 0 to 380 (median, 30) before treatment and from 0 to 300 (median, 8) after treatment. There was a significant reduction in Tregs in letrozole and letrozole cyclophosphamide patients (P < 0.0001 and P < 0.002, respectively) after treatment. Treg number at residual histology was inversely related with response (P < 0.03 and P = 0.50, respectively) and a greater Treg reduction was observed in responding patients (P < 0.03). CONCLUSION: This study suggests that aromatase inhibitors may have an indirect antitumor mechanism of action through reducing Tregs in breast tumors and may be of use in estrogen receptor-alpha-negative tumors in combination with immunotherapy approaches. PMID- 19188179 TI - Increased intratumoral FOXP3-positive regulatory immune cells during interleukin 2 treatment in metastatic renal cell carcinoma. AB - PURPOSE: The administration of interleukin-2 (IL-2) may increase the frequency of peripherally circulating FOXP3-positive regulatory immune cells, thus potentially compromising this treatment option for patients with metastatic renal cell carcinoma. The impact of IL-2-based therapy on the accumulation of FOXP3-positive immune cells in the tumor microenvironment in metastatic renal cell carcinoma is unknown. EXPERIMENTAL DESIGN: Baseline (n = 58) and on-treatment (n = 42) tumor core biopsies were prospectively obtained from patients with clear cell metastatic renal cell carcinoma before and during IL-2-based immunotherapy. Immunohistochemical expression of FOXP3 was estimated by stereological counting technique and correlated with other immune cell subsets and overall survival. RESULTS: A significant increase in absolute intratumoral FOXP3-positive immune cells was observed comparing baseline (median 23 cells/mm2; range, 0-183) and on treatment biopsies (median, 89 cells/mm2; range, 11-388; P < 0.001). The relative increase in individual patients was median 4.7-fold, range 0.3 to 230. FOXP3 positive cells were positively correlated with CD3-positive, CD4-positive, and CD8-positive tumor-infiltrating immune cells at baseline and during treatment (P < 0.05 in all comparisons). All patients achieving high numbers (>180 cells/mm2) of on-treatment FOXP3-positive intratumoral immune cells were dead within 22 months (n = 11), whereas patients with low numbers (<180 cells/mm2) of on treatment FOXP3-positive cells (n = 31) had a 5-year survival rate of 19% (hazard ratio, 2.2; confidence interval, 1.03-4.5; P = 0.043). All long-term survivors were characterized by low-baseline FOXP3-positive cells and a modest absolute rise in FOXP3-positive cells. CONCLUSION: Intratumoral FOXP3-positive regulatory immune cells significantly increased during IL-2-based immunotherapy, and high numbers of on-treatment FOXP3-positive cells were correlated with poor prognosis in patients with metastatic renal cell carcinoma. PMID- 19188180 TI - Treatment of Philadelphia-positive chronic myeloid leukemia with imatinib: importance of a stable molecular response. AB - PURPOSE: The achievement of a major molecular response (MMolR) at 12 months is a surrogate marker of progression-free survival in chronic myeloid leukemia patients treated with imatinib. EXPERIMENTAL DESIGN: We evaluated the prognostic value of the long-term evolution of the molecular response based on a retrospective analysis of 130 late chronic phase chronic myeloid leukemia patients who achieved a complete cytogenetic response (CCgR) with 400 mg/d imatinib and have now a median follow-up of 72 months (range, 48-77). RESULTS: In 71 (55%) patients, molecular response was consistently major (stable MMolR); in 19 (15%) patients, molecular response was occasionally less than major (unstable MMolR); in 40 (30%) patients, MMolR was never achieved (never MMolR) during all the course of CCgR. Patients with stable MMolR had a longer CCgR duration and a significantly better progression-free survival compared with patients with absent or unstable MMolR. The achievement of a MMolR, if maintained continuously, conferred a marked long-term stability to the CCgR: patients with a stable MMolR have a significantly lower risk of losing the CCgR than patients with unstable and never MMolR (4% versus 21%, P = 0.03, and 4% versus 33%, P < 0.0001, respectively). Finally, if a MMolR is not maintained consistently, the risk of losing the CCgR is higher but not significantly than if it is never achieved (33% versus 21%, P = 0.5). CONCLUSIONS: These data confirm that achieving a MMolR is prognostically important but point out that the prognostic value of achieving a MMolR is greater if the response is confirmed and stable. PMID- 19188181 TI - Phase II trial of Gliadel plus O6-benzylguanine in adults with recurrent glioblastoma multiforme. AB - PURPOSE: This phase II trial was designed to define the efficacy of Gliadel wafers in combination with an infusion of O6-benzylguanine (O6-BG) that suppresses tumor O6-alkylguanine-DNA alkyltransferase (AGT) levels in patients with recurrent glioblastoma multiforme for 5 days and to evaluate the safety of this combination therapy. EXPERIMENTAL DESIGN: This was a phase II, open-label, single center trial. On gross total resection of the tumor, up to eight Gliadel wafers were implanted. Bolus infusion of O6-BG was administered at 120 mg/m2 over 1 hour on days 1, 3, and 5, along with a continuous infusion at 30 mg/m2/d. The primary end points were 6-month overall survival (OS) and safety, and the secondary end points were 1-year, 2-year, and median OS. RESULTS: Fifty-two patients were accrued. The 6-month OS was 82% [95% confidence interval (95% CI), 72-93%]. The 1- and 2-year OS rates were 47% (95% CI, 35-63%) and 10% (95% CI, 3 32%), respectively. The median OS was 50.3 weeks (95% CI, 36.1-69.4 weeks). Treatment-related toxicity with this drug combination included grade 3 hydrocephalus (9.6%), grade 3 cerebrospinal fluid (CSF) leak (19.2%), and grade 3 CSF/brain infection (13.4%). CONCLUSION: The efficacy of implanted Gliadel wafers may be improved with the addition of O6-BG. Although systemically administered O6 BG can be coadministered with Gliadel wafers safely, it may increase the risk of hydrocephalus, CSF leak, and CSF/brain infection. Future trials are required to verify that inhibition of tumor AGT levels by O6-BG results in increased efficacy of Gliadel wafers without added toxicity. PMID- 19188182 TI - A phase I study of samarium lexidronam/bortezomib combination therapy for the treatment of relapsed or refractory multiple myeloma. AB - PURPOSE: This open-label, phase I dose-escalation study assessed the safety, tolerability, and initial efficacy of Samariam 153 (153Sm)-lexidronam/bortezomib combination therapy for patients with relapsed/refractory multiple myeloma. EXPERIMENTAL DESIGN: Patients were enrolled in six cohorts and given bortezomib (1.0 or 1.3 mg/m2) on days 1, 4, 8, and 11 and 153Sm-lexidronam (0.25, 0.5, or 1.0 mCi/kg) on day 3 of a 56-day cycle (maximum of four cycles). The primary endpoints were safety and tolerability of the 153Sm-lexidronam/bortezomib regimen. RESULTS: Twenty-four patients were enrolled. Median values for age, time since diagnosis, and number of prior treatments were 63 years, 29 months, and three regimens, respectively. The most common toxicities were hematologic; during the first cycle, median neutrophil and platelet nadirs were 1,000/mm3 and 98,500/mm3, respectively, and observed generally 3 to 4 weeks post-treatment. The incidences of grade 4 neutropenia and thrombocytopenia were 12.5% and 8.3%, respectively, during treatment cycle 1. Dose-limiting toxicity, reached in cohort 6 as a result of hematologic toxicity, defined the maximum tolerated dose as 0.5 mCi/kg 153Sm-lexidronam in combination with 1.3 mg/m2 bortezomib. The maximum tolerated dose for 153Sm-lexidronam in combination with the 1.0 mg/m2 bortezomib was not reached. No nonhematologic dose-limiting toxicities were observed; both the incidence and the severity of peripheral neuropathy were low. Responses occurred in 5 (21%) patients, including 3 (12.5%) complete and 2 (8.3%) minimal responses. CONCLUSIONS: Bortezomib combined with 153Sm-lexidronam appears to be a well-tolerated regimen, which showed clinical activity in this phase I trial for patients with relapsed or refractory multiple myeloma. PMID- 19188184 TI - Induction of p53-specific immunity by a p53 synthetic long peptide vaccine in patients treated for metastatic colorectal cancer. AB - PURPOSE: The tumor-associated self-antigen p53 is commonly overexpressed in cancer, including colorectal cancer, and can serve as a target for immunotherapy. The safety and immunogenicity of a p53 synthetic long peptide (p53-SLP) vaccine were investigated in patients treated for metastatic colorectal cancer. EXPERIMENTAL DESIGN: Ten patients were vaccinated twice with a set of 10 overlapping p53-SLP in a phase I/II trial. Both the safety and the breadth, magnitude, and polarization of vaccine-induced p53-specific T cells was evaluated in blood samples drawn before and after vaccination by IFN-gamma enzyme-linked immunospot, proliferation, cytokine secretion, and multiparameter flow cytometry. The migratory capacity of p53-specific T cells was evaluated by assessing their presence in a biopsy of the second vaccination site. RESULTS: Toxicity was limited to grade 1/2, mostly at the vaccination site. p53-specific T-cell responses were induced in 9 of 10 colorectal cancer patients as measured by IFN gamma enzyme-linked immunospot, proliferation, and cytokine bead array. In 6 of 9 tested patients, p53-specific T-cell reactivity persisted at least 6 months. Furthermore, p53-specific T cells isolated from the vaccination site were characterized as CD4+ T cells producing both T-helper types 1 and 2 cytokines on stimulation with p53 peptide and p53 protein. Multiparameter flow cytometry revealed that only a minor population of the p53-specific CD4+ T cells was optimally polarized. CONCLUSIONS: The p53-SLP vaccine is safe and capable to induce p53-specific T-cell responses in patients treated for colorectal cancer. New trials should focus on improving the polarization of the p53-SLP vaccine induced T-cell response. PMID- 19188185 TI - Response to sunitinib malate in advanced alveolar soft part sarcoma. AB - PURPOSE: Alveolar soft part sarcoma (ASPS) is a rare, chemoresistant soft tissue sarcoma. ASPS harbors the t(17-X) (p11.2;q25) translocation, resulting in the ASPACR1-TFE3 fusion protein, causing MET autophosphorylation and activation of downstream signaling. The tumor vascular pattern prompted us to use sunitinib malate (SM), a tyrosine kinase inhibitor with antiangiogenic properties. EXPERIMENTAL DESIGN: Since July 2007, five patients with progressive metastatic ASPS have been treated with continuous SM 37.5 mg/d on a named basis. Four patients are evaluable for response. In four cases, cryopreserved material was available. Upstream and downstream targets of receptor tyrosine kinase (RTK) pathways, as well as mechanisms of activation, were investigated by biochemical profiles, including human phospho-receptor RTK antibody arrays and immunoprecipitation/Western blotting, molecular analyses, immunohistochemistry, and fluorescence in situ hybridization analyses. RESULTS: After 3 months, two patients had RECIST (response evaluation criteria in solid tumor) partial response, as well as positron emission tomography response and subjective improvement. One had a RECIST stable disease. One progressed and stopped treatment. One patient is still responding after 12 months. The upstream analysis showed activation of all the platelet-derived growth factor receptor (PDGFR) family members, as well as epidermal growth factor receptor, MET families, and RET. Vascular endothelial growth factor receptors (VEGFR1 and VEGFR2) were activated only in one case. The downstream target analysis showed strong activation of phosphatidylinositol 3-kinase/AKT, extracellular signal-regulated kinase 1/2, and mTOR and its targets (S6K and S6). The absence of any upstream mTOR effector deregulation and the presence of RTK cognate ligands support an autocrine-paracrine activation loop mechanism. CONCLUSION: SM may have antitumor activity in ASPS, possibly through a mechanism involving PDGFR and RET. The role of MET, epidermal growth factor receptor, and mTOR, as well as PDGFR inhibition, needs to be further explored. PMID- 19188183 TI - Expression of sorafenib targets in melanoma patients treated with carboplatin, paclitaxel and sorafenib. AB - BACKGROUND: Sorafenib, a multitarget kinase inhibitor, inhibits members of the mitogen-activated protein kinase (MAPK) pathway and receptor tyrosine kinases, including vascular endothelial growth factor receptor 2 (VEGF-R2). Sorafenib, carboplatin, and paclitaxel (SCP) has antitumor activity in melanoma patients, but no association was found between response and activating B-Raf V600E mutations. We assessed the expression of sorafenib targets in SCP-treated patient specimens and evaluated the association with response and progression-free survival. EXPERIMENTAL DESIGN: Using automated quantitative analysis, we quantified the expression of VEGF-R1, VEGF-R2, VEGF-R3, fibroblast growth factor receptor 1, platelet-derived growth factor receptor beta, c-Kit, B-Raf, C-Raf, meiosis-specific serine/threonine protein kinase 1, and extracellular regulated kinase 1/2 (ERK1/2) in pretreatment specimens from 46 patients. Furthermore, we assessed ERK1/2 expression in 429 archival melanomas. RESULTS: VEGF-R2 expression was significantly higher in patients with a complete or partial response (P = 0.0435), whereas ERK1/2 was higher in patients who did not respond (P = 0.0417). High ERK1/2 was an independent predictor of poor survival. High ERK1/2 was associated with decreased survival in the archival melanoma cohort, suggesting that high ERK1/2-expressing tumors are biologically more aggressive. All of the six patients with both high VEGF-R2 and low ERK1/2 responded to SCP. CONCLUSIONS: High VEGF-R2 expression is associated with response to SCP in melanoma, whereas high ERK1/2 is associated with resistance. Collection of specimens from SCP treated melanoma patients in a cooperative group phase III trial comparing this regimen with the chemotherapy alone is ongoing, and confirmation of these findings is necessary. These markers might be useful for predicting response to sorafenib when given with other chemotherapies and in other diseases, resulting in the possible elimination of unnecessary treatment of patients unlikely to respond. PMID- 19188186 TI - Genetic variants and family history predict prostate cancer similar to prostate specific antigen. AB - PURPOSE: Although prostate-specific antigen (PSA) is the best biomarker for predicting prostate cancer, its predictive performance needs to be improved. Results from the Prostate Cancer Prevention Trial revealed the overall performance measured by the areas under curve of the receiver operating characteristic at 0.68. The goal of the present study is to assess the ability of genetic variants as a PSA-independent method to predict prostate cancer risk. EXPERIMENTAL DESIGN: We systematically evaluated all prostate cancer risk variants that were identified from genome-wide association studies during the past year in a large population-based prostate cancer case-control study population in Sweden, including 2,893 prostate cancer patients and 1,781 men without prostate cancer. RESULTS: Twelve single nucleotide polymorphisms were independently associated with prostate cancer risk in this Swedish study population. Using a cutoff of any 11 risk alleles or family history, the sensitivity and specificity for predicting prostate cancer were 0.25 and 0.86, respectively. The overall predictive performance of prostate cancer using genetic variants, family history, and age, measured by areas under curve was 0.65 (95% confidence interval, 0.63-0.66), significantly improved over that of family history and age (0.61%; 95% confidence interval, 0.59-0.62; P = 2.3 x 10(-10)). CONCLUSION: The predictive performance for prostate cancer using genetic variants and family history is similar to that of PSA. The utility of genetic testing, alone and in combination with PSA levels, should be evaluated in large studies such as the European Randomized Study for Prostate Cancer trial and Prostate Cancer Prevention Trial. PMID- 19188189 TI - Ethical responsibility of phase 0 trials. PMID- 19188187 TI - Associations of high-grade prostate cancer with BRCA1 and BRCA2 founder mutations. AB - PURPOSE: Protein-truncating mutations in BRCA1 and in particular BRCA2 genes have been associated with prostate cancer. However, there is still uncertainty about the magnitude of association particularly with Gleason score, and family history of prostate, breast, and ovary cancers. EXPERIMENTAL DESIGN: To further examine associations between three founder mutations located in BRCA1 (185delAG, 5382insC) or BRCA2 (6174delT) genes and prostate cancer, we conducted a study of 979 prostate cancer cases and 1,251 controls among Ashkenazi Jewish men. Detailed information was obtained on prostate cancer pathology, age at diagnosis, and family history of all cancers. Odds ratios (OR) and 95% confidence intervals (CIs) were estimated using logistic regression models. RESULTS: Prostate cancer risk was increased (OR, 1.9; 95% CI 0.9-4.1) for BRCA2 mutation carriers but not for BRCA1 mutation carriers. BRCA2 mutation carriers had an OR of 3.2 (95% CI, 1.4-7.3) for Gleason score of 7 to 10, but no association was observed for Gleason score of < 7. Carriers of BRCA1-185delAG mutation also had an OR of 3.5 (95% CI, 1.2-10.3) for Gleason score of > or =7 tumors; however, the association of either BRCA1-185delAG or 5382insC mutation was not statistically significant. Associations between founder mutations and prostate cancer were stronger in men with no first-degree family history of breast and/or ovarian cancers but were unaffected by family history of prostate cancer. CONCLUSION: These results indicate that the BRCA2 founder mutation confers a 3-fold elevated risk of high grade prostate cancer. Although BRCA1 mutations were not associated with prostate cancer, the BRCA1-185delAG was associated with high Gleason score tumors. These findings should be carefully considered in genetic counseling and/or evaluating therapeutic options. PMID- 19188190 TI - Benchmarking regulatory network reconstruction with GRENDEL. AB - MOTIVATION: Over the past decade, the prospect of inferring networks of gene regulation from high-throughput experimental data has received a great deal of attention. In contrast to the massive effort that has gone into automated deconvolution of biological networks, relatively little effort has been invested in benchmarking the proposed algorithms. The rate at which new network inference methods are being proposed far outpaces our ability to objectively evaluate and compare them. This is largely due to a lack of fully understood biological networks to use as gold standards. RESULTS: We have developed the most realistic system to date that generates synthetic regulatory networks for benchmarking reconstruction algorithms. The improved biological realism of our benchmark leads to conclusions about the relative accuracies of reconstruction algorithms that are significantly different from those obtained with A-BIOCHEM, an established in silico benchmark. AVAILABILITY: The synthetic benchmark utility and the specific benchmark networks that were used in our analyses are available at http://mblab.wustl.edu/software/grendel/. PMID- 19188191 TI - SciMiner: web-based literature mining tool for target identification and functional enrichment analysis. AB - SciMiner is a web-based literature mining and functional analysis tool that identifies genes and proteins using a context specific analysis of MEDLINE abstracts and full texts. SciMiner accepts a free text query (PubMed Entrez search) or a list of PubMed identifiers as input. SciMiner uses both regular expression patterns and dictionaries of gene symbols and names compiled from multiple sources. Ambiguous acronyms are resolved by a scoring scheme based on the co-occurrence of acronyms and corresponding description terms, which incorporates optional user-defined filters. Functional enrichment analyses are used to identify highly relevant targets (genes and proteins), GO (Gene Ontology) terms, MeSH (Medical Subject Headings) terms, pathways and protein-protein interaction networks by comparing identified targets from one search result with those from other searches or to the full HGNC [HUGO (Human Genome Organization) Gene Nomenclature Committee] gene set. The performance of gene/protein name identification was evaluated using the BioCreAtIvE (Critical Assessment of Information Extraction systems in Biology) version 2 (Year 2006) Gene Normalization Task as a gold standard. SciMiner achieved 87.1% recall, 71.3% precision and 75.8% F-measure. SciMiner's literature mining performance coupled with functional enrichment analyses provides an efficient platform for retrieval and summary of rich biological information from corpora of users' interests. AVAILABILITY: http://jdrf.neurology.med.umich.edu/SciMiner/. A server version of the SciMiner is also available for download and enables users to utilize their institution's journal subscriptions. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 19188192 TI - Accurate identification of orthologous segments among multiple genomes. AB - MOTIVATION: The accurate detection of orthologous segments (also referred to as syntenic segments) plays a key role in comparative genomics, as it is useful for inferring genome rearrangement scenarios and computing whole-genome alignments. Although a number of algorithms for detecting orthologous segments have been proposed, none of them contain a framework for optimizing their parameter values. METHODS: In the present study, we propose an algorithm, named OSfinder (Orthologous Segment finder), which uses a novel scoring scheme based on stochastic models. OSfinder takes as input the positions of short homologous regions (also referred to as anchors) and explicitly discriminates orthologous anchors from non-orthologous anchors by using Markov chain models which represent respective geometric distributions of lengths of orthologous and non-orthologous anchors. Such stochastic modeling makes it possible to optimize parameter values by maximizing the likelihood of the input dataset, and to automate the setting of the optimal parameter values. RESULTS: We validated the accuracies of orthology mapping algorithms on the basis of their consistency with the orthology annotation of genes. Our evaluation tests using mammalian and bacterial genomes demonstrated that OSfinder shows higher accuracy than previous algorithms. AVAILABILITY: The OSfinder software was implemented as a C++ program. The software is freely available at http://osfinder.dna.bio.keio.ac.jp under the GNU General Public License. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 19188193 TI - High-performance gene name normalization with GeNo. AB - MOTIVATION: The recognition and normalization of textual mentions of gene and protein names is both particularly important and challenging. Its importance lies in the fact that they constitute the crucial conceptual entities in biomedicine. Their recognition and normalization remains a challenging task because of widespread gene name ambiguities within species, across species, with common English words and with medical sublanguage terms. RESULTS: We present GeNo, a highly competitive system for gene name normalization, which obtains an F-measure performance of 86.4% (precision: 87.8%, recall: 85.0%) on the BioCreAtIvE-II test set, thus being on a par with the best system on that task. Our system tackles the complex gene normalization problem by employing a carefully crafted suite of symbolic and statistical methods, and by fully relying on publicly available software and data resources, including extensive background knowledge based on semantic profiling. A major goal of our work is to present GeNo's architecture in a lucid and perspicuous way to pave the way to full reproducibility of our results. AVAILABILITY: GeNo, including its underlying resources, will be available from www.julielab.de. It is also currently deployed in the Semedico search engine at www.semedico.org. PMID- 19188194 TI - The natural soluble form of IL-18 receptor beta exacerbates collagen-induced arthritis via modulation of T-cell immune responses. AB - OBJECTIVE: IL-18 is a pluripotent cytokine that has been implicated in the development of rheumatoid arthritis. A soluble form of the IL-18 receptor accessory protein (sIL-18Rbeta) with unknown function has recently been identified. This study examined the ability of sIL-18Rbeta to inhibit IL-18 biological activities and to modulate immune responses during collagen-induced arthritis (CIA). METHODS: Adenoviruses encoding sIL-18Rbeta were administered intravenously in type II collagen-immunised DBA/1 mice. Humoral responses were analysed by determining anti-bovine collagen type II (BCII) antibody levels by ELISA. Cytokine production by splenic T cells and cytokine levels in serum were measured by Luminex multi-analyte technology. CD4(+)CD25(+)Foxp3(+) regulatory T cells (Treg) were measured by flow cytometry. RESULTS: Intravenous delivery of Ad5.sIL-18Rbeta in collagen-immunised mice led to enhanced transgene expression in splenic antigen-presenting cells (APC). A co-culture of these sIL-18Rbeta transduced APC with purified splenic CD3(+) T cells led to a marked inhibition of IL-18-induced IFNgamma, IL-4 and IL-17 production by CD3(+) T cells. Remarkably, systemic treatment with Ad5.sIL-18Rbeta caused an exacerbation of arthritis, and histological evaluation of knee joints showed increased cartilage and bone erosion. No significant differences were observed in anti-BCII antibodies, but the aggravation was accompanied by decreased IFNgamma (-30%) and IL-4 (-44%) and increased IL-17 (+84%) production by splenic CD3(+) T cells. In addition, reduced circulating levels of CD4(+)CD25(+)Foxp3(+) Treg and anti-inflammatory IL-10 were shown. CONCLUSION: This study identifies sIL-18Rbeta as a novel IL-18 inhibitor, which promotes CIA after intravenous overexpression by affecting Treg levels and supporting a T helper type 17 response. PMID- 19188195 TI - Solar maculopathy in a young child. PMID- 19188196 TI - Effect of village income and household income on sanitation facilities, hygiene behaviours and child undernutrition during rapid economic growth in a rural cross border area, Yunnan, China. AB - BACKGROUND: The study aimed to examine the effects of village income and household income on child nutrition status through basic sanitation and hygiene behaviours. METHODS: A survey was conducted in a rural cross-border area of Yunnan, China. Data on village income in 2002-2006 and household income in 2002 2007 were obtained from an official report and a household survey respectively. Anthropometric measurement of the children aged 6 months to 5 years (n = 1801) was used to determine their nutrition status. Child caretakers were interviewed about household sanitation facilities and their hygiene behaviours using a structured questionnaire. RESULTS: Households with incomes below the national poverty line decreased from 22% in 2002 to less than 8% in 2007. The coverage of safe drinking water and water-sealed latrines gradually increased, but was still inadequate. The prevalence of stunting and underweight in children was 37% and 17.5% respectively. Village income had a greater positive effect than household income on exclusive breastfeeding, drinking boiled water, handwashing with soap, as well as reducing the prevalence of stunting. Village income at one lag year had the greatest effect on the availability of basic sanitation compared with other lag years, while household income had a small but significant effect through all lag years. CONCLUSIONS: Rapid economic growth is not always followed by improved child nutrition status. Village income has a greater effect than household income on sanitation facilities, hygiene behaviours of caretakers and child nutrition status. PMID- 19188197 TI - HIV prone occupational exposures: epidemiology and factors associated with initiation of post-exposure prophylaxis. AB - BACKGROUND: Occupational exposures to bloodborne viruses are very common. Whilst occupational HIV transmissions are rare, the serious physical, psychological and cost implications of potential transmission make this an important public health topic. European and UK guidelines recommend HIV post-exposure prophylaxis (PEP) as a valuable tool of preventing occupational HIV infection. Yet one in five UK healthcare workers did not initiate PEP despite having been exposed to an HIV positive source patient. The aim of the study is to examine factors associated with PEP uptake behaviour. METHODS: The study is based on an analysis of the UK Health Protection Agency surveillance database of 'Significant Occupational Exposures to Bloodborne Viruses in Healthcare Workers'. Associations between possible predicting factors and PEP-uptake have been examined with univariate analysis and logistic regression modelling. RESULTS: Univariate analysis and logistic regression found significant associations between PEP-uptake and visible blood on the device (p<0.0001) and a linear relationship with increasing injury depth (p<0.0001). Doctors were significantly more likely to start PEP than nurses (OR 1.88, 1.16; 3.02). Multiple imputation of missing values did not significantly alter these results. CONCLUSIONS: PEP-uptake was associated with known transmission risk factors, suggesting awareness of current guidelines. The significant differences in PEP-uptake across occupation categories may be due to differential risk perceptions or other underlying factors. This is the first national study to examine PEP-uptake following occupational exposures to HIV. Further research and exploration of these findings are warranted, to understand the role of PEP-uptake behaviour in preventing occupational HIV transmission. PMID- 19188198 TI - Infantile cardiomyopathy caused by a mutation in the overlapping region of mitochondrial ATPase 6 and 8 genes. AB - BACKGROUND: Infantile cardiomyopathy is a genetically heterogeneous disorder with significant morbidity and mortality. METHODS: This study aimed to identify the mutation present in four unrelated patients who presented as infants with isolated hypertrophic cardiomyopathy. RESULTS: In all four, a novel mitochondrial m.8528T-->C mutation was identified. This results in a change of the initiation codon in ATPase 6 to threonine and a concurrent change from a highly conserved hydrophobic amino acid, tryptophan, at position 55 of ATPase 8 to a highly basic arginine. To our knowledge, this is the first report of a mutation affecting both mitochondrial genome-encoded complex V subunit proteins. Testing of the relatives of one patient indicated that the mutation is heteroplasmic and correlated with disease. CONCLUSION: Mitochondrial genome sequencing should be considered in patients with infantile hypertrophic cardiomyopathy. PMID- 19188199 TI - Risk of hypertension from exposure to road traffic noise in a population-based sample. AB - OBJECTIVES: To assess the association between hypertension and traffic noise. METHODS: The prevalence and incidence of hypertension were examined in a Swedish municipality partly affected by noise from a highway (20,000 vehicles/24 h) and a railway (200 trains/24 h). A-weighed 24 h average sound levels (L(Aeq,24h)) from road and railway traffic were calculated at each residential building using a geographical information system and a validated model. Physician-diagnosed hypertension, antihypertensive medication and background factors were evaluated in 1953 individuals using postal questionnaires (71% response rate). Prevalence ratios and odds ratios (ORs) were calculated for different noise categories. Based on year of moving into the residence and year of diagnosis, person-years and incidence rates of hypertension were estimated, as well as relative risks including covariates, using Poisson and Cox regression. RESULTS: When road traffic noise, age, sex, heredity and body mass index were included in logistic regression models, and allowing for >10 years of latency, the OR for hypertension was 1.9 (95% CI 1.1 to 3.5) in the highest noise category (56-70 dBA) and 3.8 (95% CI 1.6 to 9.0) in men. The incidence rate ratio was increased in this group of men, and the relative risk of hypertension in a Poisson regression model was 2.9 (95% CI 1.4 to 6.2). There were no clear associations in women or for railway noise. CONCLUSIONS: The study shows a positive association between residential road traffic noise and hypertension among men, and an exposure-response relationship. While prevalence ratios were increased, findings were more pronounced when incidence was assessed. PMID- 19188200 TI - Workplace social capital and co-occurrence of lifestyle risk factors: the Finnish Public Sector Study. AB - OBJECTIVE: The aim of this prospective study was to examine the link between individual and ecological workplace social capital and the co-occurrence of adverse lifestyle risk factors such as smoking, heavy drinking, physical inactivity and overweight. METHODS: Data on 25 897 female and 5476 male public sector employees were analysed. Questionnaire surveys conducted in 2000-2002 (baseline) and 2004-2005 (follow-up) were used to assess workplace social capital, lifestyle risk factors and other characteristics. Multilevel multinomial logistic regression analysis was used to examine associations between individual and ecological social capital and the co-occurrence of lifestyle risk factors. RESULTS: In the cross-sectional analysis adjusted for age, sex, marital status and employer, low social capital at work at both the individual and ecological level was associated with at least a 1.3 times higher odds of having more than two lifestyle risk factors versus having no risk factors. Similar associations were found in the prospective setting. However, additional adjustment for the co occurrence of risk factors and socioeconomic status at baseline attenuated the result to non-significant. CONCLUSION: Social capital at work seems to be associated with a lowered risk of co-occurrence of multiple lifestyle risk factors but does not clearly predict the future risk of this co-occurrence. PMID- 19188201 TI - Respiratory and skin effects of exposure to wood dust from the rubber tree Hevea brasiliensis. AB - OBJECTIVES: Potential health effects related to wood dust from the rubber tree, which produces natural rubber latex, have not been previously investigated. The main aim of this study was to investigate the relations of rubber tree dust exposure to respiratory and skin symptoms, asthma and lung function. METHODS: A cross-sectional study was conducted among 103 workers (response rate 89%) in a rubber tree furniture factory and 76 office workers (73%) in four factories in Thailand. All participants answered a questionnaire and performed spirometry. Inhalable dust levels were measured in different work areas. RESULTS: Factory workers showed increased risk of wheezing, nasal symptoms and asthma compared to office workers. There was a dose-dependent increase in wheeze and skin symptoms in relation to dust level. Significantly increased risks of nasal symptoms (adj OR 3.67, 95% CI 1.45 to 9.28) and asthma (8.41, 1.06 to 66.60) were detected in the low exposure category. Workers exposed to ethyl cyanoacrylate glue had significantly increased risk of cough, breathlessness and nasal symptoms. There was dose-dependent reduction in spirometric lung function with wood dust level. CONCLUSIONS: This study provides new evidence that workers exposed to wood dust from the rubber tree experience increased risk of nasal symptoms, wheeze, asthma and skin symptoms and have reduced spirometric lung function. Exposure to cyanoacrylate is related to significantly increased respiratory symptoms. Results suggest that the furniture industry using rubber tree wood should implement appropriate exposure control measures to reduce wood dust exposure and cyanoacrylate glue exposure to protect their employees. PMID- 19188202 TI - Anticontagionism between 1821 and 1867: The Fielding H. Garrison Lecture. 1948. PMID- 19188203 TI - Commentary: Ackerknecht and 'Anticontagionism': a tale of two dichotomies. PMID- 19188204 TI - Commentary: Epidemiology in context. PMID- 19188205 TI - Commentary: Disease etiology and political ideology: revisiting Erwin H Ackerknecht's Classic 1948 Essay, 'Anticontagionism between 1821 and 1867'. PMID- 19188206 TI - Commentary: Light drinking in pregnancy: can a glass or two hurt? PMID- 19188207 TI - Estimates of measles case fatality ratios: a comprehensive review of community based studies. AB - BACKGROUND: Global deaths from measles have decreased notably in past decades, due to both increases in immunization rates and decreases in measles case fatality ratios (CFRs). While some aspects of the reduction in measles mortality can be monitored through increases in immunization coverage, estimating the level of measles deaths (in absolute terms) is problematic, particularly since incidence-based methods of estimation rely on accurate measures of measles CFRs. These ratios vary widely by geographic and epidemiologic context and even within the same community from year-to-year. METHODS: To understand better the variations in CFRs, we reviewed community-based studies published between 1980 and 2008 reporting age-specific measles CFRs. RESULTS: The results of the search consistently document that measles CFRs are highest in unvaccinated children under age 5 years; in outbreaks; the lowest CFRs occur in vaccinated children regardless of setting. The broad range of case and death definitions, study populations and geography highlight the complexities in extrapolating results for global public health planning. CONCLUSIONS: Values for measles CFRs remain imprecise, resulting in continued uncertainty about the actual toll measles exacts. PMID- 19188208 TI - Age at menarche, total mortality and mortality from ischaemic heart disease and stroke: the Adventist Health Study, 1976-88. AB - BACKGROUND: Little is known about the relationship between age at menarche and total mortality and mortality from ischaemic heart disease and stroke. METHODS: A cohort study of 19 462 Californian Seventh-Day Adventist women followed-up from 1976 to 1988. A total of 3313 deaths occurred during follow-up, of which 809 were due to ischaemic heart disease and 378 due to stroke. RESULTS: An early menarche was associated with increased total mortality (P-value for linear trend <0.001), ischaemic heart disease (P-value for linear trend = 0.01) and stroke (P-value for linear trend = 0.02) mortality. There were, however, also some indications of an increased ischaemic heart disease mortality in women aged 16-18 at menarche (5% of the women). When assessed as a linear relationship, a 1-year delay in menarche was associated with 4.5% (95% CI 2.3-6.7) lower total mortality. The association was stronger for ischaemic heart disease [6.0% (95% CI 1.2-10.6)] and stroke [8.6% (95% CI 1.6-15.1)] mortality. CONCLUSIONS: The results suggest that there is a linear, inverse relationship between age at menarche and total mortality as well as with ischaemic heart disease and stroke mortality. PMID- 19188209 TI - Geographical information systems as a tool for monitoring tobacco industry advertising. AB - BACKGROUND: Although the use of a geographical information systems (GIS) approach is usually applied to epidemiological disease outbreaks and environmental exposure mapping, it has significant potential as a tobacco control research tool in monitoring point-of-purchase (POP) tobacco advertising. DESIGN: An ecological study design approach was applied so as to primarily evaluate and interpret the spatial density and intensity of POP and tobacco industry advertisements within <300 m to high schools in Greece with the application of GIS methodology combining mapping, photographing and global positioning data. RESULTS: The GIS approach identified 133 POP and 44 billboards within 300 m of the school gates of Heraklion schools. On average 13 POP (range 4-21) and 4.4 billboards (range 1-9) were located per school, and all had at least 1 POP within 20 m of the school gate. On average (SD) 9 (6) tobacco advertisements per POP (range 0-25) were noted, and 80% of them were below child height. The GIS protocol identified that kiosks, that were excepted from the Greek ban on tobacco advertising, in comparison to other POP, were found not only to be closer and visible from the school gates (44.1% vs 10.8%, p<0.001) but were also found to have more external advertisements (8 (5) vs 5 (3), p<0.001). CONCLUSIONS: This study demonstrates the effectiveness of a GIS system in monitoring tobacco industry advertising on a large population-based scale and implies its use as a standardised method for monitoring tobacco industry strategies and tobacco control efforts. PMID- 19188210 TI - Tobacco in prisons: a focus group study. AB - OBJECTIVE: To examine the role of tobacco use in prison and possible influences of the prison environment on smoking among inmates in the context of developing inmate smoking cessation programmes. METHOD: Qualitative study based on seven focus groups with prisoners and ex-prisoners. SETTINGS: A maximum security prison in rural New South Wales (NSW), Australia, and a community justice restorative centre and accommodation service for ex-prisoners in Sydney, NSW, Australia. PARTICIPANTS: 40 participants (28 men and 12 women) comprising nine prisoners (including four Indigenous inmates) and 31 ex-prisoners. RESULTS: Prisoners reported that tobacco serves as a de facto currency in correctional settings and can be exchanged for goods, used to pay debts and for gambling. Smoking helps manage the stressful situations such as transfers, court appearances and prison visits. Inmate smoking cessation programmes need to address the enmeshment of tobacco in prison life, improve availability of pharmacotherapies (for example, nicotine patches, bupropion) and the quitline (a free telephone helpline providing information on stopping smoking), provide non-smoking cells and areas within prisons, encourage physical activity for inmates and maintain monitoring of smoking cessation status after release. CONCLUSIONS: Tobacco is integrally bound up in the prison "culture". Our findings are relevant to inform prison health authorities concerned with improving the health of prisoners, and for support organisations attempting to facilitate smoking cessation both in prison and after release. Smoking cessation programmes in prisons should be tailored to the unique stresses of the prison environment. Programmes need to acknowledge the difficulties of quitting smoking in prison arising from the stresses posed by this setting. PMID- 19188211 TI - Associations between migrant status and sexually transmitted infections among female sex workers in Tijuana, Mexico. AB - OBJECTIVE: To examine associations between migration and sexually transmitted infection (STI) prevalence among Mexican female sex workers (FSW). METHODS: FSW aged 18 years and older in Tijuana, Baja California (BC) underwent interviews and testing for HIV, syphilis, gonorrhoea and chlamydia. Multivariate logistic regressions identified correlates of STI. RESULTS: Of 471 FSW, 79% were migrants to BC. Among migrant FSW, prevalence of HIV, syphilis, gonorrhoea, chlamydia and any STI was 6.6%, 13.2%, 7.8%, 16.3% and 31.1% compared with 10.9%, 18.2%, 13.0%, 19.0% and 42.4% among FSW born in BC. A greater proportion of migrant FSW were registered with local health services and were ever tested for HIV. Migrant status was protective for any STI in unadjusted models (unadjusted odds ratio 0.61, 95% CI 0.39 to 0.97). In multivariate models controlling for confounders, migrant status was not associated with an elevated odds of STI acquisition and trended towards a protective association. CONCLUSIONS: Unexpectedly, migrant status (vs native-born status) appeared protective for any STI acquisition. It is unclear which social or economic conditions may protect against STI and whether these erode over time in migrants. Additional research is needed to inform our understanding of whether or how geography, variations in health capital, or social network composition and information-sharing attributes can contribute to health protective behaviours in migrant FSW. By capitalising on such mechanisms, efforts to preserve protective health behaviours in migrant FSW will help control STI in the population and may lead to the identification of strategies that are generalisable to other FSW. PMID- 19188212 TI - Embolism of the pulmonary artery stump after right pneumonectomy. AB - We report a case of a 53-year-old male who presented with thoracodynia three months after right pneumonectomy. Chest CT-scan demonstrated thrombus at the pulmonary artery stump without any other abnormal finding. He was treated successfully with acenocoumarol. We present this case analyzing the possible causes and discussing the treatment. PMID- 19188213 TI - Risk factors for low colloid osmotic pressure during infant cardiopulmonary bypass with a colloidal prime. AB - Extensive variations of colloid osmotic pressure (COP) measured in the priming as well as during infant cardiopulmonary bypass motivated us to audit clinical and laboratory data to identify the risk factors for low COP at the end of bypass. Data of 73 consecutive infant patients with body weight <10 kg, who underwent elective, first time open-heart surgery between March 2005 and December 2006 were examined. The following variables were analyzed: COP, blood loss, transfusion requirements and hematological data. Univariate and multivariate analysis of risk factors for low COP (<15 mmHg) was performed. Forty-eight percent of patients had COP <15 mmHg at the end of bypass. Those patients had significantly lower COP before start of bypass, during, and at the end of the operation. Significant univariate predictors of low COP at the end of bypass were: lower patient weight; lower COP before start of bypass, lower priming COP and larger volume of cardioplegia received into the circulation. After multivariable analysis, lower patient COP before bypass remained the only significant predictor for low COP at the end of bypass. Pre-bypass crystalloid dilution during induction should be avoided, as this is the most important cause of low COP during the bypass. Priming COP and COP management strategy should be adapted to the individual patient demand. PMID- 19188215 TI - Conflicting recommendations. Checklist for guidelines. PMID- 19188214 TI - Outcome of depression in later life in primary care: longitudinal cohort study with three years' follow-up. AB - OBJECTIVES: To study the duration of depression, recovery over time, and predictors of prognosis in an older cohort (>or=55 years) in primary care. DESIGN: Longitudinal cohort study, with three years' follow-up. SETTING: 32 general practices in West Friesland, the Netherlands. PARTICIPANTS: 234 patients aged 55 years or more with a prevalent major depressive disorder. MAIN OUTCOME MEASURES: Depression at baseline and every six months using structured diagnostic interviews (primary care evaluation of mental disorders according to diagnoses in Diagnostic and Statistical Manual of Mental Disorders, fourth edition) and a measure of severity of symptoms (Montgomery Asberg depression rating scale). The main outcome measures were time to recovery and the likelihood of recovery at different time points. Multivariable analyses were used to identify variables predicting prognosis. RESULTS: The median duration of a major depressive episode was 18.0 months (95% confidence interval 12.8 to 23.1). 35% of depressed patients recovered within one year, 60% within two years, and 68% within three years. A poor outcome was associated with severity of depression at baseline, a family history of depression, and poorer physical functioning. During follow-up functional status remained limited in patients with chronic depression but not in those who had recovered. CONCLUSION: Depression among patients aged 55 years or more in primary care has a poor prognosis. Using readily available prognostic factors (for example, severity of the index episode, a family history of depression, and functional decline) could help direct treatment to those at highest risk of a poor prognosis. PMID- 19188216 TI - Do patients' preferences matter?. Patients' preference and mental capacity. PMID- 19188217 TI - Promotion of exercise by GPs. Results not strong enough for GPs to implement BMJ study. PMID- 19188218 TI - Patient reported trial outcomes. What about the cost to use the measures? PMID- 19188219 TI - Obesity and pregnancy. Obesity is a risk factor for thromboembolism. PMID- 19188220 TI - ECT's increasing evidence base and positive profile. PMID- 19188221 TI - Print formats for BMJ research. Different strokes may be needed for different folks. PMID- 19188222 TI - Print formats for BMJ research. BMJ pico--a window into the future? PMID- 19188223 TI - Fish oil and arrhythmias. Pro-arrhythmic effects of fish oils. PMID- 19188224 TI - GP performance website. A waste of time and money. PMID- 19188225 TI - Patients' attendance at clinics is worse with choose and book. PMID- 19188226 TI - Into the therapeutics void. Let us protest loudly. PMID- 19188227 TI - Into the therapeutics void. A lecturer from BSMS explains. PMID- 19188228 TI - Into the therapeutics void. A student from BSMS writes. PMID- 19188229 TI - Conflicting recommendations. Let's not forget AGREE. PMID- 19188230 TI - Case of Baby P. There are no "basics" in child protection. PMID- 19188233 TI - Number of breast cancer cases detected by screening has doubled in a decade in England. PMID- 19188235 TI - Obama drops funding from bill, alarming family planning groups. PMID- 19188236 TI - Health department sets up cost sharing deal for multiple myeloma drug. PMID- 19188237 TI - Female doctors are more likely to follow heart failure guidelines than male counterparts. PMID- 19188238 TI - Open access to MRI for general practitioners: 12 years' experience at one institution -- a retrospective analysis. AB - The aim of this study was to evaluate 12 years of general practitioner (GP) use of open access MRI services at a single London teaching hospital. A retrospective analysis of reports from all GP requests for MRI scans between 1994 and 2005 was performed. The date, scanned body part, and requester details from 1798 scans requested by 209 individual GPs over a continuous 12-year period were recorded. All scans were then graded into four categories based on the severity of reported findings from normal to gross abnormality. Over the study period, GP requests as a percentage of the total (MRI) department workload remained low at approximately 2.6%. Spine, knee and brain requests constituted 86% (n = 1546) of requested scans. 48% (n = 868) of scans were reported as normal or minor degenerative changes only. 26% (n = 466) of scans demonstrated serious pathology that was likely to warrant hospital consultant referral. There was a wide range of scans requested per requester, from 1 to 240 over the period, with an average of 8.5 scans per GP. In conclusion, any department wishing to set up open access to MRI services for GPs could cover the majority of requests by offering spine, knee and brain imaging. The percentage of normal report rates for GP requests is comparable with previous studies of outpatient referrals. A large variation in requesting patterns between GPs suggests the need for increased communication between GPs and imaging departments to optimise use of the service. PMID- 19188239 TI - Three-day regimen improves faecal tagging for minimal preparation CT examination of the colon. AB - This study set out to determine whether extending the length of oral contrast administration in minimal preparation CT of the colon improves faecal tagging. Two cohorts of 50 patients each were compared, one with a 2-day the other with a 3-day faecal tagging regimen. The degree of faecal tagging was graded by two blinded observers. The 3-day regimen showed significantly better tagging in the rectum and sigmoid colon (p = 0.006 and p = 0.009, respectively, using the Mann Whitney test). The percentage of patients who had faecal tagging in the sigmoid colon graded as "complete" was 64% for the 3-day regimen as opposed to 34% for the 2-day regimen. The corresponding percentages for the rectum were 64% for the 3-day regimen and 36% for the 2-day regimen. Extending the length of oral contrast administration from 2 to 3 days significantly improves the quality of faecal tagging in the rectum and sigmoid colon. PMID- 19188240 TI - The effect of treatment position, prone or supine, on dose-volume histograms for pelvic radiotherapy in patients with rectal cancer. AB - Patients undergoing radiotherapy for rectal cancer are generally treated in a prone position, with a full bladder, to reduce the volume of normal bowel in the high-dose volume. This position is difficult to maintain, and is not consistently reproducible. This study evaluates the volume of bowel and dose received in the prone and supine positions in patients undergoing pre-operative rectal cancer chemoradiation. Using CT planning, 19 consecutive patients with rectal cancer with a full bladder underwent CT scanning first in the prone position and then immediately afterwards in the supine position. The planning target volume was outlined for the prone position and transcribed to the supine scan using pre-set criteria. The bladder and small bowel were outlined in both positions. Radiotherapy was planned using three-dimensional conformal planning, and treatment was delivered using three fields with multileaf collimators in two phases: phase I, pelvis 45 Gy/25 fractions; and phase II, tumour 9 Gy/five fractions. For both positions, the volume of bowel receiving doses in 5 Gy increments from 5-45 Gy was calculated using dose-volume histograms. At 5 Gy and 10 Gy dose levels, a significantly higher volume of bowel was irradiated in the supine position (p<0.001). At 15 Gy, it was marginally significant (p = 0.018). From 20-45 Gy, there was no significant difference in the volume of bowel irradiated with each 5 Gy increment. This study demonstrates that the volume of bowel irradiated at doses associated with bowel toxicity in concurrent chemoradiation is not significantly higher in the supine position. This position could be adopted for patients undergoing pre-operative rectal cancer chemoradiation. PMID- 19188241 TI - Relationship between the extent of deep venous thrombosis and the extent of acute pulmonary embolism as assessed by CT angiography. AB - The aim of our study is to investigate prospectively the quantitative relationship between deep venous thrombosis (DVT) and acute pulmonary embolism (PE). 110 patients clinically suspected of having venous thromboembolic disease underwent combined CT pulmonary angiography (CTPA) and venography of lower limb veins. 44 patients presented with clinical signs of DVT and positive ultrasonography or ascending venography, but no clinical sign of PE (Group 1). 66 patients presented with clinical signs of PE and positive CTPA (Group 2). Clot load in lower limb veins and pulmonary arteries were scored by two independent readers, each using two separate systems for DVT and two for PE. 27 (61%) patients in Group 1 also had PE, and 55 (83%) patients in Group 2 also had DVT. Correlations between PE and DVT scores were weak but statistically significant in Group 2 (r(s) ranging from 0.470-0.520; p< or =0.001), but only some were significant in Group 1 (r(s) ranging from 0.253-0.318; p-values ranging from 0.035-0.097). In conclusion, although PE occurs in a majority of patients with DVT, and vice versa, the amount/burden of clot load in one condition does not necessarily indicate - or indicates only weakly - the degree of burden in the other condition. PMID- 19188242 TI - Radiation dose measurements for personnel performing 90Y-ibritumomab tiuxetan administration: a comparison between two injection methods for dose reduction. AB - The purpose of this study was to directly measure, using thermoluminescent dosimeters, the radiation doses received by radiation team members performing (90)Y-ibritumomab tiuxetan administration. The occupational doses associated with two injection methods for patient administration - an automatic syringe driver and an injection box - were compared. The associated risks, namely cancer induction and hereditary effect, were also estimated from the results and compared with risk factors recommended by the International Commission on Radiological Protection publication 103. The results showed that the doses received by the index and thumb of the right hand and the index finger of the left hand of the radiation oncologist were significantly reduced by using the injection box method. The difference in the dose received by the medical physicist using the two methods was not statistically significant. It was observed that three pairs of latex gloves could further reduce the dose to the hands. The radiological risks of cancer induction and hereditary effect were negligible: of the order of 10(-6) and 10(-7) per (90)Y-ibritumomab tiuxetan administration, respectively, for both methods. However, the results of our study also showed that it would be possible in a busy centre for pregnant women to receive a dose of (90)Y-ibritumomab tiuxetan that exceeds the recommended annual dose limit for the surface of the abdomen. PMID- 19188243 TI - Adult resuscitation: are we up to date? A study of staff resuscitation skills in the radiology department of a tertiary referral centre. AB - This study was undertaken to determine the current level of resuscitation skills amongst staff in the Department of Clinical Radiology, University Hospital of Wales, and to identify ways of improving it. Questionnaires, which were modified according to expected level of expertise, were distributed to all staff members. 66% of the staff responded. 75% had formal resuscitation training, but the validation of this training had lapsed in 66%. 11% were aware of the location of all the resuscitation equipment in the department. Only 10% were aware of the new Resuscitation Council guidelines (changed in April 2006) regarding chest compression to ventilation ratio and hand position during chest compression. Only 57% of the medical staff and radiographers could identify and manage an anaphylactic reaction. Only 55% of medical staff could identify and manage a pneumothorax; and correctly measure and insert an oropharyngeal airway. 35% could use a defibrillator, but only 6% were aware of the changes to the guidelines for use of this equipment. Only one staff member was aware of the all the relevant changes in the guidelines. There is a shortfall of resuscitation skills in the radiology department and a responsibility on all radiology staff to update their resuscitation skills. PMID- 19188244 TI - Liposoluble antioxidants provide an effective radioprotective barrier. AB - Ionising radiation causes the massive generation of reactive oxygen species and induces cellular DNA damage. The antioxidant, protective effects of several compounds against gamma-ray-induced chromosomal damage were determined by the micronucleus test, evaluating the reduction in the frequency of micronuclei in cytokinesis-blocked human lymphocytes. The compounds studied were added to human blood at 25 microM, 5 min before or after irradiation with 2 Gy of caesium-137. The results suggest that different protective mechanisms are operating in each case. When the phenolic compounds are added before gamma-irradiation, their protective antimutagenic activity is based on their scavenging capacity against superoxide anion (O(2)(.-)) and, especially, hydroxyl radical ((.)OH), regardless of whether they are oil- or water-soluble compounds. When the phenolic compounds are added after gamma-irradiation treatment, the protective effect relies on activity against reactive oxygen species present in cells, i.e. lipoperoxy radicals (R(-)OO(.)), which are mainly responsible for continuous chromosomal oxidative damage. In addition, ionising radiation enhances lysosomal enzyme secretion and arachidonate release from membranes through lipo-oxygenase, cyclo oxygenase and phospholipase activities, thus increasing the inflammatory cell response. Only oil-soluble compounds, such as carnosic acid, carnosol and delta tocopherol, provide a significant protective antimutagenic activity. The most powerful water-soluble antioxidants lack the capacity to protect against gamma ray-induced damage. The difference between anti-radical and anti-lipoperoxidant activities could explain the different behaviour of the compounds tested in terms of protecting against the lipid peroxidative processes. This anti-lipoperoxidant activity depends on several factors, but it is clear that only the lipo antioxidants are effective in protecting human cells against oxidative damage, even when administered after exposure to ionising radiation. PMID- 19188245 TI - A novel approach for reduction of brown fat uptake on FDG PET. AB - The aim of this study was to determine whether or not a single dose of a beta blocker, such as propranolol 40 mg, administered 60 min prior to (18)F fluorodeoxyglucose ((18)F-FDG) injection would help reduce brown fat uptake of (18)F-FDG. Patients who were referred for either a pre-treatment or a post treatment evaluation positron emission tomography (PET) scan and who showed (18)F FDG uptake in brown adipose tissue (BAT) were included in this study. The total number of patients who showed uptake in BAT and in whom a repeat study was carried out after propranolol injection was 40. A repeat PET scan was carried out after an interval of at least 48 h. Propranolol at a dose of 40 mg was given orally 60 min prior to the (18)F-FDG injection. A whole-body PET scan was performed on a dedicated whole-body PET scanner (ADVANCE, GE Medical Systems, Milwaukee, WI), using attenuation correction with 68-Ge external pin sources. We observed that (18)F-FDG uptake in BAT was absent in 36 (90%) patients post propranolol. We conclude that propranolol reduces the uptake of (18)F-FDG in BAT, and thus improves the accuracy of PET imaging. PMID- 19188246 TI - Effect of tube current on diagnostic image quality in paediatric cerebral multidetector CT images. AB - The aim of this study was to investigate the effect of tube current on diagnostic image quality in paediatric cerebral multidetector CT (MDCT) images in order to identify the minimum radiation dose required to reproduce acceptable levels of different diagnostic image qualities. Original digital scanning data (raw data) were selected retrospectively from routine MDCT brain examinations of 25 paediatric patients. All examinations had been performed using axial scanning on an eight-slice MDCT (LightSpeed Ultra, GE Healthcare). Their ages ranged from newborn to 15 years. Quantum noise was added artificially to the raw data representing dose reductions equivalent to steps of 20 mA. Patient identification information was removed. Three experienced radiologists blindly and randomly assessed the resulting images from two different levels of the brain with regard to reproduction of structures and overall image quality. Final data were evaluated using the non-parametric statistical approach of inter-scale concordance. The minimum value of tube current-time product (mAs) required to reproduce an image of sufficient diagnostic quality was established in relation to the age of the patient. The corresponding CT dose index values by volume (CTDI(vol) (mGy)) were also established. In conclusion, acceptable reproduction of low-contrast structures was possible at CTDI(vol) values down to 20 mGy (patients 1-5 years old). For acceptable reproduction of high-contrast structures, CTDI(vol) values down to 10 mGy were considered possible (patients 1 5 years old). The original image quality for patients under 6 months of age (15 mGy) was found to be inadequate for acceptable reproduction of low-contrast structures. PMID- 19188247 TI - Activity-dependent development of inhibitory synapses and innervation pattern: role of GABA signalling and beyond. AB - GABA-mediated synaptic inhibition is crucial in neural circuit operations. The development of GABAergic inhibitory synapses and innervation pattern in mammalian neocortex is a prolonged process, extending well into the postnatal period, and is regulated by neural activity and experience. Accumulating evidence supports the hypothesis that GABA signalling acts beyond synaptic transmission and regulates inhibitory synapse development; in other words, similar to glutamate signalling at developing excitatory synapses, GABA may coordinate pre- and post synaptic maturation at inhibitory synapses. These findings raise numerous questions regarding the underlying mechanisms, including the role of GABA receptors and their link to synaptic adhesion molecules. Since synapse formation is a crucial component of axon growth, GABA signalling may also shape the axon arbor and innervation pattern of inhibitory neurons. A mechanism unique to GABAergic neurons is activity-dependent GABA synthesis, largely mediated through activity-regulated transcription of the rate-limiting enzyme GAD67. Such cell wide as well as synaptic regulation of GABA signalling may constitute a mechanism by which input levels and patterns onto GABAergic neurons shape their innervation pattern during circuit development. PMID- 19188248 TI - A Ca(2+)-calmodulin-eEF2K-eEF2 signalling cascade, but not AMPK, contributes to the suppression of skeletal muscle protein synthesis during contractions. AB - Skeletal muscle protein synthesis rate decreases during contractions but the underlying regulatory mechanisms are poorly understood. It was hypothesized that there would be a coordinated regulation of eukaryotic elongation factor 2 (eEF2) and eukaryotic initiation factor 4E-binding protein 1 (4EBP1) phosphorylation by signalling cascades downstream of rises in intracellular [Ca(2+)] and decreased energy charge via AMP-activated protein kinase (AMPK) in contracting skeletal muscle. When fast-twitch skeletal muscles were contracted ex vivo using different protocols, the suppression of protein synthesis correlated more closely with changes in eEF2 than 4EBP1 phosphorylation. Using a combination of Ca(2+) release agents and ATPase inhibitors it was shown that the 60-70% suppression of fast twitch skeletal muscle protein synthesis during contraction was equally distributed between Ca(2+) and energy turnover-related mechanisms. Furthermore, eEF2 kinase (eEF2K) inhibition completely blunted increases in eEF2 phosphorylation and partially blunted (i.e. 30-40%) the suppression of protein synthesis during contractions. The 3- to 5-fold increase in skeletal muscle eEF2 phosphorylation during contractions in situ was rapid and sustained and restricted to working muscle. The increase in eEF2 phosphorylation and eEF2K activation were downstream of Ca(2+)-calmodulin (CaM) but not other putative activating factors such as a fall in intracellular pH or phosphorylation by protein kinases. Furthermore, blunted protein synthesis and 4EBP1 dephosphorylation were unrelated to AMPK activity during contractions, which was exemplified by normal blunting of protein synthesis during contractions in muscles overexpressing kinase-dead AMPK. In summary, in fast-twitch skeletal muscle, the inhibition of eEF2 activity by phosphorylation downstream of Ca(2+) CaM-eEF2K signalling partially contributes to the suppression of protein synthesis during exercise/contractions. PMID- 19188249 TI - The hungry fetus? Role of leptin as a nutritional signal before birth. AB - In adult animals, leptin is an adipose-derived hormone that is important primarily in the regulation of energy balance during short- and long-term changes in nutritional state. Expression of leptin and its receptors is widespread in fetal and placental tissues, although the role of leptin as a nutritional signal in utero is unclear. Before birth, leptin concentration correlates with several indices of fetal growth, and may be an endocrine marker of fetal size and energy stores in the control of metabolism and maturation of fetal tissues. In addition, leptin synthesis and plasma concentration can be modified by insulin, glucocorticoids, thyroid hormones and oxygen availability in utero, and therefore, leptin may be part of the hormonal response to changes in the intrauterine environment. Evidence is emerging to show that leptin has actions before birth that are tissue-specific and may occur in critical periods of development. Some of these actions are involved in the growth and development of the fetus and others have long-term consequences for the control of energy balance in adult life. PMID- 19188250 TI - Bradykinin-induced astrocyte-neuron signalling: glutamate release is mediated by ROS-activated volume-sensitive outwardly rectifying anion channels. AB - Glial cells release gliotransmitters which signal to adjacent neurons and glial cells. Previous studies showed that in response to stimulation with bradykinin, glutamate is released from rat astrocytes and causes NMDA receptor-mediated elevation of intracellular Ca(2+) in adjacent neurons. Here, we investigate how bradykinin-induced glutamate release from mouse astrocytes signals to neighbouring neurons in co-cultures. Astrocyte-to-neuron signalling and bradykinin-induced glutamate release from mouse astrocytes were both inhibited by the anion channel blocker 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS) and phloretin. Glutamate release was also sensitive to 4-(2-Butyl-6,7 dichlor-2-cyclopentylindan-1-on-5-yl) oxybutyric acid (DCPIB), a specific blocker of the volume-sensitive outwardly rectifying anion channel (VSOR). Astrocytes, but not neurons, responded to bradykinin with activation of whole-cell Cl- currents. Although astrocytes stimulated with bradykinin did not undergo cell swelling, the bradykinin-activated current exhibited properties typical of VSOR: outward rectification, inhibition by osmotic shrinkage, sensitivity to DIDS, phloretin and DCPIB, dependence on intracellular ATP, and permeability to glutamate. Bradykinin increased intracellular reactive oxygen species (ROS) in mouse astrocytes. Pretreatment of mouse astrocytes with either a ROS scavenger or an NAD(P)H oxidase inhibitor blocked bradykinin-induced activation of VSOR, glutamate release and astrocyte-to-neuron signalling. By contrast, pretreatment with BAPTA-AM or tetanus neurotoxin A failed to suppress bradykinin-induced glutamate release. Thus, VSOR activated by ROS in mouse astrocytes in response to stimulation with bradykinin, serves as the pathway for glutamate release to mediate astrocyte-to-neuron signalling. Since bradykinin is an initial mediator of inflammation, VSOR might play a role in glia-neuron communication in the brain during inflammation. PMID- 19188251 TI - Properties of distinct ventral tegmental area synapses activated via pedunculopontine or ventral tegmental area stimulation in vitro. AB - Anatomical studies indicate that synaptic inputs from many cortical and subcortical structures converge on neurons of the ventral tegmental area (VTA). Although in vitro electrophysiological studies have examined synaptic inputs to dopamine (DA) and non-DA neurons in the VTA, they have largely relied upon local electrical stimulation to activate these synapses. This provides little information regarding the distinct properties of synapses originating from different brain areas. Using whole-cell recordings in parasagittal rat brain slices that preserved subcortical axons from the pedunculopontine nucleus (PPN) to the VTA, we compared these synapses with those activated by intra-VTA stimulation. PPN-evoked currents demonstrated longer latencies than intra-VTA evoked currents, and both VTA and PPN responses were mediated by GABA(A) and AMPA receptors. However, unlike VTA-evoked currents, PPN currents were exclusively mediated by glutamate in 25-40% of the VTA neurons. Consistent with a cholinergic projection from the PPN to the VTA, nicotinic acetylcholine receptors (nAChR) were activated by endogenous acetylcholine released during PPN, but not VTA, stimulation. This was seen as a reduction of PPN-evoked, and not VTA-evoked, synaptic currents by the alpha7-nAChR antagonist methyllycaconitine (MLA) and the agonist nicotine. The beta2-nAChR subunit antagonist dihydro-beta-erythroidine had no effect on VTA- or PPN-evoked synaptic currents. The effects of MLA on PPN evoked currents were unchanged by the GABA(A) receptor blocker picrotoxin, indicating that alpha7-nAChRs presynaptically modulated glutamate and not GABA release. These differences in physiological and pharmacological properties demonstrate that ascending PPN and presumed descending inputs to VTA utilize distinct mechanisms to differentially modulate neuronal activity and encode cortical and subcortical information. PMID- 19188252 TI - Rapamycin administration in humans blocks the contraction-induced increase in skeletal muscle protein synthesis. AB - Muscle protein synthesis and mTORC1 signalling are concurrently stimulated following muscle contraction in humans. In an effort to determine whether mTORC1 signalling is essential for regulating muscle protein synthesis in humans, we treated subjects with a potent mTORC1 inhibitor (rapamycin) prior to performing a series of high-intensity muscle contractions. Here we show that rapamycin treatment blocks the early (1-2 h) acute contraction-induced increase ( approximately 40%) in human muscle protein synthesis. In addition, several downstream components of the mTORC1 signalling pathway were also blunted or blocked by rapamycin. For instance, S6K1 phosphorylation (Thr421/Ser424) was increased post-exercise 6-fold in the control group while being unchanged with rapamycin treatment. Furthermore, eEF2 phosphorylation (Thr56) was reduced by approximately 25% post-exercise in the control group but phosphorylation following rapamycin treatment was unaltered, indicating that translation elongation was inhibited. Rapamycin administration prior to exercise also reduced the ability of raptor to associate with mTORC1 during post-exercise recovery. Surprisingly, rapamycin treatment prior to resistance exercise completely blocked the contraction-induced increase in the phosphorylation of ERK1/2 (Thr202/Tyr204) and blunted the increase in MNK1 (Thr197/202) phosphorylation. However, the phosphorylation of a known target of MNK1, eIF4E (Ser208), was similar in both groups (P > 0.05) which is consistent with the notion that rapamycin does not directly inhibit MAPK signalling. We conclude that mTORC1 signalling is, in part, playing a key role in regulating the contraction-induced stimulation of muscle protein synthesis in humans, while dual activation of mTORC1 and ERK1/2 stimulation may be required for full stimulation of human skeletal muscle protein synthesis. PMID- 19188253 TI - Effect of intracellular lipid droplets on cytosolic Ca2+ and cell death during ischaemia-reperfusion injury in cardiomyocytes. AB - Lipid droplets (LD) consist of accumulations of triacylglycerols and have been proposed to be markers of ischaemic but viable tissue. Previous studies have described the presence of LD in myocardium surviving an acute coronary occlusion. We investigated whether LD may be protective against cell death secondary to ischaemia-reperfusion injury. The addition of oleate-bovine serum albumin complex to freshly isolated adult rat cardiomyocytes or to HL-1 cells resulted in the accumulation of intracellular LD detectable by fluorescence microscopy, flow cytometry and (1)H-nuclear magnetic resonance spectroscopy. Simulated ischaemia reperfusion of HL-1 cells (respiratory inhibition at pH 6.4 followed by 30 min of reperfusion) resulted in significant cell death (29.7+/-2.6% of total lactate dehydrogenase release). However, cell death was significantly attenuated in cells containing LD (40% reduction in LDH release compared with control cells, P=0.02). The magnitude of LD accumulation was inversely correlated (r(2)=0.68, P=0.0003) with cell death. The protection associated with intracellular LD was not a direct effect of the fatty acids used to induce their formation, because oleate added 30 min before ischaemia, during ischaemia or during reperfusion did not form LD and did not protect against cell death. Increasing the concentration of free oleate during reperfusion progressively decreased the protection afforded by LD. HL-1 cells labelled with fluo-4, a Ca(2+)-sensitive fluorochrome, fluorescence within LD areas increased more throughout simulated ischaemia and reperfusion than in the cytosolic LD-free areas of the same cells. As a consequence, cells with LD showed less cytosolic Ca(2+) overload than control cells. These results suggest that LD exert a protective effect during ischaemia-reperfusion by sequestering free fatty acids and Ca(2+). PMID- 19188254 TI - Sem1p and Ubp6p orchestrate telomeric silencing by modulating histone H2B ubiquitination and H3 acetylation. AB - Histone covalent modifications and 26S proteasome-mediated proteolysis modulate many regulatory events in eukaryotes. In Saccharomyces cerevisiae, heterochromatin mediates transcriptional silencing at telomeres, HM loci and rDNA array. Here, we show that proteasome-associated Sem1p and its interacting partner, Ubp6p (a deubiquitinating enzyme), are essential to maintain telomeric silencing. Simultaneous deletion of SEM1 and UBP6 induces dramatic silencing defect accompanied by significantly increased level of ubiquitinated-histone H2B and markedly reduced levels of acetylated-lysine 14 and 23 on histone H3 at the telomeres. Further, the loss of Sem1p and Ubp6p triggers relocation of silencing factors (e.g. Sir proteins) from telomere to HM loci and rDNA array. Such relocation of silencing factors enhances gene silencing at HM loci and rDNA array, but diminishes telomeric silencing. Interestingly, both Sem1p and Ubp6p participate in the proteolytic function of the proteasome. However, we find that the telomeric silencing is not influenced by proteolysis. Taken together, our data demonstrate that Sem1p and Ubp6p maintain telomeric heterochromatin structure (and hence silencing) through modulation of histone covalent modifications and association of silencing factors independently of the proteolytic function of the proteasome, thus offering a new regulatory mechanism of telomeric silencing. PMID- 19188255 TI - Effects of magnesium and related divalent metal ions in topoisomerase structure and function. AB - The catalytic steps through which DNA topoisomerases produce their biological effects and the interference of drug molecules with the enzyme-DNA cleavage complex have been thoroughly investigated both from the biophysical and the biochemical point of view. This provides the basic structural insight on how this family of essential enzymes works in living systems and how their functions can be impaired by natural and synthetic compounds. Besides other factors, the physiological environment is known to affect substantially the biological properties of topoisomerases, a key role being played by metal ion cofactors, especially divalent ions (Mg(2+)), that are crucial to bestow and modulate catalytic activity by exploiting distinctive chemical features such as ionic size, hardness and characteristics of the coordination sphere including coordination number and geometry. Indeed, metal ions mediate fundamental aspects of the topoisomerase-driven transphosphorylation process by affecting the kinetics of the forward and the reverse steps and by modifying the enzyme conformation and flexibility. Of particular interest in type IA and type II enzymes are ionic interactions involving the Toprim fold, a protein domain conserved through evolution that contains a number of acidic residues essential for catalysis. A general two-metal ion mechanism is widely accepted to account for the biophysical and biochemical data thus far available. PMID- 19188256 TI - Adenylation of plant miRNAs. AB - The modification or degradation of RNAs including miRNAs may play vital roles in regulating RNA functions. The polyadenylation- and exosome-mediated RNA decay is involved in the degradation of plant RNAs including the primary miRNA processing intermediates. However, plant miRNA levels are not affected by exosome depletion. Here, we report the cloning of a large number of 5' and/or 3' truncated versions of the known miRNAs from various tissues of Populus trichocarpa (black cottonwood). It suggests that plant miRNAs may be degraded through either 5' to 3' or 3' to 5' exonucleolytic digestion. We also show that a significant portion of the isolated miRNAs contains, at the 3'-end, one or a few post transcriptionally added adenylic acid residues, which are distinct in length from the polyadenylate tail added to other plant RNAs for exosome-mediated degradation. Using an in vitro miRNA degradation system, where synthesized miRNA oligos were degraded in extracts of P. trichocarpa cells, we revealed that the adenylated miRNAs were degraded slower than others without adenylation. It indicates that addition of adenylic acid residues on the 3'-end plays a negative role in miRNA degradation. Our results provide new information for understanding the mechanism of miRNA degradation. PMID- 19188257 TI - Model-based redesign of global transcription regulation. AB - Synthetic biology aims to the design or redesign of biological systems. In particular, one possible goal could be the rewiring of the transcription regulation network by exchanging the endogenous promoters. To achieve this objective, we have adapted current methods to the inference of a model based on ordinary differential equations that is able to predict the network response after a major change in its topology. Our procedure utilizes microarray data for training. We have experimentally validated our inferred global regulatory model in Escherichia coli by predicting transcriptomic profiles under new perturbations. We have also tested our methodology in silico by providing accurate predictions of the underlying networks from expression data generated with artificial genomes. In addition, we have shown the predictive power of our methodology by obtaining the gene profile in experimental redesigns of the E. coli genome, where rewiring the transcriptional network by means of knockouts of master regulators or by upregulating transcription factors controlled by different promoters. Our approach is compatible with most network inference methods, allowing to explore computationally future genome-wide redesign experiments in synthetic biology. PMID- 19188258 TI - Human DNA polymerase theta possesses 5'-dRP lyase activity and functions in single-nucleotide base excision repair in vitro. AB - DNA polymerase theta (Pol theta) is a low-fidelity DNA polymerase that belongs to the family A polymerases and has been proposed to play a role in somatic hypermutation. Pol theta has the ability to conduct translesion DNA synthesis opposite an AP site or thymine glycol, and it was recently proposed to be involved in base excision repair (BER) of DNA damage. Here, we show that Pol theta has intrinsic 5'-deoxyribose phosphate (5'-dRP) lyase activity that is involved in single-nucleotide base excision DNA repair (SN-BER). Full-length human Pol theta is a approximately 300-kDa polypeptide, but we show here that the 98-kDa C-terminal region of Pol theta possesses both DNA polymerase activity and dRP lyase activity and is sufficient to carry out base excision repair in vitro. The 5'-dRP lyase activity is independent of the polymerase activity, in that a polymerase inactive mutant retained full 5'-dRP lyase activity. Domain mapping of the 98-kDa enzyme by limited proteolysis and NaBH(4) cross-linking with a BER intermediate revealed that the dRP lyase active site resides in a 24-kDa domain of Pol theta. These results are consistent with a role of Pol theta in BER. PMID- 19188259 TI - AID can restrict L1 retrotransposition suggesting a dual role in innate and adaptive immunity. AB - Retrotransposons make up over 40% of the mammalian genome. Some copies are still capable of mobilizing and new insertions promote genetic variation. Several members of the APOBEC3 family of DNA cytosine deaminases function to limit the replication of a variety of retroelements, such as the long-terminal repeat (LTR) containing MusD and Ty1 elements, and that of the non-LTR retrotransposons, L1 and Alu. However, the APOBEC3 genes are limited to mammalian lineages, whereas retrotransposons are far more widespread. This raises the question of what cellular factors control retroelement transposition in species that lack APOBEC3 genes. A strong phylogenetic case can be made that an ancestral activation induced deaminase (AID)-like gene duplicated and diverged to root the APOBEC3 lineage in mammals. Therefore, we tested the hypothesis that present-day AID proteins possess anti-retroelement activity. We found that AID can inhibit the retrotransposition of L1 through a DNA deamination-independent mechanism. This mechanism may manifest in the cytoplasmic compartment co- or posttranslationally. Together with evidence for AID expression in the ovary, our data combined to suggest that AID has innate immune functions in addition to its integral roles in creating antibody diversity. PMID- 19188260 TI - A cellulose synthase-containing compartment moves rapidly beneath sites of secondary wall synthesis. AB - The woody secondary walls of plants represent the major sites of cellulose deposition. The polymerization of cellulose occurs at the plasma membrane by the secondary wall cellulose synthase complex (CSC). In the long, cylindrical cells that make up the xylem, secondary wall deposition is confined to discrete regions of the cell, and yellow fluorescent protein (YFP)-labeled CSCs are also localized to these regions. Using fluorescence loss in photobleaching (FLIP) of complete hoops containing YFP-CSCs, we demonstrate movement of the complexes beneath the nascent secondary wall in developing xylem vessels. We have devised a method for determining particle velocities for particles moving around a cylindrical object using data from FLIP. By applying this method to the hoops of YFP-CSCs of the developing vessels, we have obtained the first estimates of speed of these complexes. These speeds are calculated to be in excess of 7 microm s(-1) and are far higher than those speeds previously reported for the primary wall complex. These high speeds are unlikely to be consistent with CSC movement being attributed to cellulose synthesis alone, and suggest the existence of a highly motile compartment beneath the nascent secondary wall. PMID- 19188261 TI - Characterization of phosphatidylinositol phosphate kinases from the moss Physcomitrella patens: PpPIPK1 and PpPIPK2. AB - Phosphoinositides (PIs) play a major role in eukaryotic cells, despite being a minor component of most membranes. This is the first report on PI metabolism in a bryophyte, the moss Physcomitrella patens. Moss PI composition is similar to that of other land plants growing under normal conditions. In contrast to the large number of PIPK genes present in flowering plants, the P. patens genome encodes only two type I/II PIPK genes, PpPIPK1 and PpPIPK2, which are very similar at both the nucleotide and protein product levels. However, the expression of the two genes is differentially regulated, and in vitro biochemical characterization shows that the resulting enzymes have different substrate specificities. PpPIPK1 uses PtdIns4P and PtdIns3P with similar preference and also metabolizes PtdIns(3,4)P(2) to produce PtdIns(3,4,5)P(3), a PI not yet detected in intact plant cells. PpPIPK2 prefers PtdIns as substrate and is much less active towards PtdIns4P and PtdIns3P. Thus, PpPIPK2 shows properties reminiscent of both PtdInsP kinase and PtdIns-kinases. Moreover, a substitution of glutamic acid by alanine in the activation loop drastically reduced PpPIPK1 activity and altered the substrate specificity to PtdIns5P being the preferred substrate compared with PtdIns4P and PtdIns3P. These findings demonstrate that the substrate specificity of plant PIPKs is determined in a plant-specific manner, which provides new insights into the regulatory modes of PIPK activity in plants. PMID- 19188262 TI - ARGONAUTE1 acts in Arabidopsis root radial pattern formation independently of the SHR/SCR pathway. AB - The formation of radially symmetric tissue patterns is one of the most basic processes in the development of vascular plants. In Arabidopsis thaliana, plant specific GRAS-type transcription factors, SHORT-ROOT (SHR) and SCARECROW (SCR), are required for asymmetric cell divisions that separate two ground tissue cell layers, the endodermis and cortex, as well as for endodermal cell fate specification. While loss of SHR or SCR results in a single-layered ground tissue, radially symmetric cellular patterns are still maintained, suggesting that unknown regulatory mechanisms act independently of the SHR/SCR-dependent pathway. In this study, we identified a novel root radial pattern mutant and found that it is a new argonaute1 (ago1) allele. Multiple ago1 mutant alleles contained supernumerary ground tissue cell layers lacking a concentric organization, while expression patterns of SHR and SCR were not affected, revealing a previously unreported role for AGO1 in root ground tissue patterning. Analyses of ago1 scr double mutants demonstrated that the simultaneous loss of the two pathways caused a dramatic reduction in cellular organization and ground tissue identity as compared with the single mutants. Based on these results, we propose that highly symmetric root ground tissue patterns are maintained by the actions of two independent pathways, one using post-transcriptional regulation mediated by AGO1 and the other using the SHR/SCR transcriptional regulator. PMID- 19188263 TI - The correlation of evolutionary rate with pathway position in plant terpenoid biosynthesis. AB - Genes are expected to face stronger selective constraint and to evolve more slowly if they encode enzymes upstream as opposed to downstream in metabolic pathways, because upstream genes are more pleiotropic, being required for a wider range of end products. However, few clear examples of this trend in evolutionary rate variation exist. We examined whether genes involved in plant terpenoid biosynthesis exhibit such a pattern, using data for 40 genes from five fully sequenced angiosperms, Oryza, Vitis, Arabidopsis, Populus, and Ricinus. Our results show that d(N)/d(S) does in fact correlate with pathway position along pathways converting glucose to the terpenoid phytohormones abscissic acid, gibberellic acid (GA), and brassinosteroids. Upstream versus downstream rate variation is particularly strong in the GA pathway. In contrast, we found no or little apparent variation in d(N)/d(S) with gene copy number. We also introduce a new measure of pathway position, the Pathway Pleiotropy Index (PPI), which counts groups of enzymes between pathway branch points. We found that this measure is superior to pathway position in explaining variation in d(N)/d(S) along each pathway. Although at least 8 of the 40 genes showed evidence of positive selection, correlations of d(N)/d(S) with PPI remain significant when these genes are removed. Therefore, our results are consistent with the prediction that selective constraint is progressively relaxed along metabolic pathways. PMID- 19188264 TI - Quantification of adaptive evolution of genes expressed in avian brain and the population size effect on the efficacy of selection. AB - Whether protein evolution is mainly due to fixation of beneficial alleles by positive selection or to random genetic drift has remained a contentious issue over the years. Here, we use two genomewide polymorphism data sets collected from chicken populations, together with divergence data from >5,000 chicken-zebra finch gene orthologs expressed in brain, to assess the amount of adaptive evolution in protein-coding genes of birds. First, we show that estimates of the fixation index (FI, the ratio of fixed nonsynonymous-to-synonymous changes over the ratio of the corresponding polymorphisms) are highly dependent on the character of the underlying data sets. Second, by using polymorphism data from high-frequency alleles, to avoid the confounding effect of slightly deleterious mutations segregating at low frequency, we estimate that about 20% of amino acid changes have been brought to fixation through positive selection during avian evolution. This estimate is intermediate to that obtained in humans (lower) and flies as well as bacteria (higher), and is consistent with population genetics theory that stipulates a positive relationship between the efficiency of selection and the effective population size. Further, by comparing the FIs for common and all alleles, we estimate that approximately 20% of nonsynonymous variation segregating in chicken populations represent slightly deleterious mutations, which is less than in Drosophila. Overall, these results highlight the link between the effective population size and positive as well as negative selection. PMID- 19188265 TI - Measurement error and model specification in determining how duration of tasks affects level of occupational exposure. AB - Statistical modeling of determinants of exposure ascertained in large-scale surveys is an increasingly popular approach to both (i) identifying effective occupational exposure controls that arise in 'natural experiments' and (ii) predicting how altering some working conditions may impact exposure levels. This paper sheds light on two underappreciated methodological challenges of such studies. First, I examine the impact of measurement error in the observed determinant of exposure on an investigator's ability to correctly rank the determinants of exposure in terms of their exposure rate (one aspect of how important a give determinant is). Simultaneously, I consider the issue of whether empirical models fitted for the sake of statistical convenience actually reflect the physical reality that is being modeled and how this may affect the answer to the question about ranking determinants of exposure. These general issues are examined in the context of the 'time per task' determinant of exposure and true exposure model that states that exposure is equal to product of exposure rate and duration of a task. Simulation studies were conducted and their conclusions applied in re-examining the data on the impact of duration of some key task on exposure levels to flour dust among bakers. The simulation study demonstrated that bias due to measurement error in observed effects can be either positive or negative. The main conclusion is that the correct ranking of exposure rates can be obtained from both true and poorly specified exposure models, but can be severely distorted by errors in estimates of the duration of tasks performed. PMID- 19188266 TI - Food protein-induced enterocolitis syndrome: 16-year experience. AB - OBJECTIVE: The goal was to examine the demographic characteristics, causative foods, clinical features, treatments, and outcomes for children presenting with acute food protein-induced enterocolitis syndrome. METHODS: This was a retrospective study of children with food protein-induced enterocolitis syndrome who presented to the Children's Hospital at Westmead (Sydney, Australia) over 16 years. RESULTS: Thirty-five children experienced 66 episodes of food protein induced enterocolitis syndrome. The mean age at initial presentation was 5.5 months. Children frequently experienced multiple episodes before a correct diagnosis was made. Twenty-nine children reacted to 1 food, and 6 reacted to 2 foods. Causative foods for the 35 children were rice (n = 14), soy (n = 12), cow's milk (n = 7), vegetables and fruits (n = 3), meats (n = 2), oats (n = 2), and fish (n = 1). In the 66 episodes, vomiting was the most common clinical feature (100%), followed by lethargy (85%), pallor (67%), and diarrhea (24%). A temperature of <36 degrees C at presentation was recorded for 24% of episodes. A platelet count of >500 x 10(9) cells per L was recorded for 63% of episodes with blood count results. Only 2 of the 19 children who presented to an emergency department with their initial reactions were discharged with correct diagnoses. Additional investigations of food protein-induced enterocolitis syndrome episodes presenting to the hospital were common, with 34% of patients undergoing abdominal imaging, 28% undergoing a septic evaluation, and 22% having a surgical consultation. Prognosis was good, with high rates of resolution for the 2 most common food triggers (ie, rice and soy) by 3 years of age. CONCLUSIONS: Misdiagnosis and delays in diagnosis for children with food protein-induced enterocolitis syndrome were common, leading many children to undergo unnecessary, often painful investigations. Decreased body temperature and thrombocytosis emerge as additional features of the syndrome. PMID- 19188267 TI - Responding to compliance changes in a lung model during manual ventilation: perhaps volume, rather than pressure, should be displayed. AB - OBJECTIVE: The standard technique for positive-pressure ventilation is to regulate the breath size by varying the pressure applied to the bag. Investigators have argued that consistency of peak inspiratory pressure is important. However, research shows that excessive tidal volume delivered with excessive pressure injures preterm lungs, which suggests that inspiratory pressure should be varied during times of changing compliance, such as resuscitation of newborns or treatment after surfactant delivery. METHODS: We modified a computerized lung model (ASL5000 [IngMar Medical, Pittsburgh, PA]) to simulate the functional residual capacity of a 3-kg neonate with apnea and programmed it to change compliance during ventilation. Forty-five professionals were blinded to randomized compliance changes while using a flow-inflating bag, a self-inflating bag, and a T-piece resuscitator. We instructed subjects to maintain a constant inflation volume, first while blinded to delivered volume and then with volume displayed, with all 3 devices. RESULTS: Subjects adapted to compliance changes by adjusting inflation pressure more effectively when delivered volume was displayed. When only pressure was displayed, sensing of compliance changes occurred only with the self-inflating bag. When volume was displayed, adjustments to compliance changes occurred with all 3 devices, although the self-inflating bag was superior. CONCLUSIONS: In this lung model, volume display permitted far better detection of compliance changes compared with display of only pressure. Devices for administration of positive-pressure ventilation should display volume rather than pressure. PMID- 19188268 TI - Brain injury and fever: hospital length of stay and cost outcomes. AB - Fever has been shown to be related to extended hospital stays in neurologically injured patients. We performed meta-analyses of the impact of fever on length of stay (LOS) in the Intensive Care Unit (ICU) and for total hospital length of stay, including all recent scholarship published since 1/1/1995 pertaining to thermoregulation of neurogenic fever and length of hospital stay. We also developed estimates of the financial impact on hospital costs. Fever was shown to have a large, statistically significant impact on both ICU and hospital LOS. For ICU LOS, combined effect size g = .88, Z = 4.24, P < .0001. For hospital LOS, g = .79, Z = 2.2, P = .0278. Mean additional ICU days = 5.7 days; mean additional hospital days = 8.5 days. We estimate that fever added an average of $17,414 in hospital cost to total hospital stays; mean $13,672 (95% Confidence Interval [CI]: $10,074, $17,270) in additional ICU costs and mean $3,742 (CI: -$1,203, $8,820) in additional routine costs. PMID- 19188269 TI - Mycobacterium avium septicemia with ARDS in a patient with diabetes mellitus and no other known immune-compromising condition. AB - A 39-year-old diabetic male sustained 4 culture-proven episodes of disseminated Mycobacterium avium complex infection over a span of 6 years. The first, second, and fourth episodes were manifested clinically by osteomyelitis. The third episode was characterized by Mycobacterium avium complex septicemia (positive blood cultures for Mycobacterium avium complex) and acute respiratory failure with features of acute respiratory distress syndrome requiring mechanical ventilation and positive end-expiratory pressure for up to 30 days. Studies for impaired immunity including human immunodeficiency virus infection, idiopathic CD4+ T lymphocytopenia syndrome, and interferon deficiency were negative. Intensive treatment of the fourth episode with amikacin, clarithromycin, rifampin, isoniazide, ethambutol, and pyrazinamide for 2 months followed by 2 years of treatment with the same regimen without amikacin resulted in apparent cure. The patient has remained free of clinical infection for 13 years. The possible role of diabetes in pathogenesis is discussed. PMID- 19188270 TI - Review of a large clinical series: intrahospital transport of critically ill patients: outcomes, timing, and patterns. AB - The purpose of this study was to analyze the relationship of intrahospital transport patterns with patient throughput and outcomes in an oncological intensive care unit. We retrospectively reviewed all patients admitted to a closed medical-surgical intensive care unit at a cancer center between January 1, 2004 and December 31, 2005. We compared the clinical characteristics and outcomes of patients with and without transport and analyzed all intrahospital transports in relation to intensive care unit occupancy, length of stay, and intensive care unit and hospital outcomes. Transport patterns were also assessed by day of week, time of day, timing of the first transport to intensive care unit admission, and destination. Transported patients (n = 413, 43.5%) had significantly higher severity of illness scores on intensive care unit admission, greater use of vasopressors and mechanical ventilation, and longer intensive care unit and hospital length of stay and higher hospital mortality than nontransported patients (n = 535, 56.5%). Multiple transports (!2) occurred in 45% of the transported patients. The number of transports was directly proportional to intensive care unit length of stay. The highest transport rates and nearly half of all first transports occurred during the first 24 hours of intensive care unit admission. Transports were most common during weekdays and on afternoon and evening hours and most frequently to the computed tomography suite. Our study shows that intrahospital transport of the critically ill is a multifaceted process with important implications for intensive care unit resource analysis, workload and throughput. PMID- 19188271 TI - Bedside insertion of inferior vena cava filters by a medical intensivist: preliminary results. AB - The development of percutaneous techniques has allowed vena cava filters to be safely placed at the bedside. Such procedures appear uniquely suited for critically ill patients as they avoid the inherent risks associated with transportation. We report a "medical intensivist's" experience with the placement of 13 percutaneous vena cava filters. Vena cava filters were successfully placed in 12 of 13 patients as confirmed by postplacement venacavograms; 1 placement attempt was aborted because of a completely occluded inferior vena cava. These results suggest that with appropriate training and supervision, medical intensivists can safely insert vena cava filters in the intensive care unit setting. Such a practice appears safe, reduces patient risk associated with intrahospital transport, and may promote cost containment. PMID- 19188272 TI - The effects of ventilatory mode on lung aeration assessed with computer tomography: a randomized controlled study. AB - Maintenance of spontaneous breathing superimposed on mechanical ventilation is suggested to improve gas exchange in patients with acute lung injury. The aim of this study was to evaluate the long-term effects of airway pressure release ventilation with maintained unsupported spontaneous breathing (APRV) and synchronized intermittent mandatory ventilation with pressure support (SIMV) on the amount of lung collapse in acute lung injury patients. Thirty-seven patients with acute lung injury were studied in a trial comparing APRV or SIMV. Computer assisted tomography scannings (CT) were performed before randomization and at day 7. The change in the amount of nonaerated lung was comparable between groups; 14.7% (3.8-17.4) in APRV group (n = 13) and 9.6% (-1.4 to 18.62) in the SIMV group (n = 10), (P = .65, difference in mean 4.9%, 95% confidence interval -9.0% to 19.0%). The effects of APRV and SIMV on lung aeration are similar after 7 days of mechanical ventilation. PMID- 19188273 TI - Recombinant activated factor VII following pediatric cardiac surgery. AB - OBJECTIVE: Review the use of recombinant activated Factor VII following cardiac surgery. Specifically, we sought to define our current therapeutic practice indications and outcomes to assess the impact of recombinant activated factor VII on postoperative bleeding. DESIGN: Retrospective case series. SETTING: The study was conducted at the University affiliated pediatric intensive care unit. PATIENTS AND PARTICIPANTS: All postcardiac surgical patients who received recombinant activated Factor VII between June 2002 and July 2006. RESULTS: Cardiac surgery requiring cardiopulmonary bypass was performed on 1010 children during this period. In all, 25 (2.5%) children received recombinant activated factor VII for excessive bleeding. A single dose (180 microg/kg) of recombinant activated factor VII was given to 11 patients and 2 doses of 180 microg/kg to 14 patients. Intercostal drain losses were reduced from 12 (6.7-20.8) mL/kg/h to 3 (1-4.1) mL/kg/h, P = .018 following 1 dose of recombinant activated factor VII. In those receiving 2 doses; initial losses were 19.1 (7.5-31.7) mL/kg/h, after the first dose were 7.5 (3.6-13.7) mL/kg/h, P = .046, and after the second dose were 2 (1-2.9) mL/kg/h, P = .008. The plasma prothrombin time decreased in both the 1 dose, P = .003 and 2 dose, P = .009 groups. The activated partial thromboplastin time also decreased in the 1 dose group, P = .007 and 2 dose group, P = .03. There were no side effects attributable to recombinant activated factor VII. Annual rates of return to the operating theatre for excessive bleeding were coincidentally reduced in association with the routine use of recombinant activated factor VII from 4.3% to 1.5%, P = .019. CONCLUSIONS: Hemostasis occurred in 25 postoperative pediatric cardiac patients after recombinant activated Factor VII was given. In this setting, once conventional hemostatic therapy was optimized, recombinant activated Factor VII 180 microg/kg initially with intercostal losses greater than 10 mL/kg/h and a repeat dose after 2 hours if losses remained greater than 5 mL/kg/h was effective. No complications were found to have occurred and there was a coincidental reduction in annual returns to theatre for excessive bleeding. PMID- 19188274 TI - Thalamic input to distal apical dendrites in neocortical layer 1 is massive and highly convergent. AB - Input to apical dendritic tufts is now deemed crucial for associative learning, attention, and similar "feedback" interactions in the cerebral cortex. Excitatory input to apical tufts in neocortical layer 1 has been traditionally assumed to be predominantly cortical, as thalamic pathways directed to this layer were regarded relatively scant and diffuse. However, the sensitive tracing methods used in the present study show that, throughout the rat neocortex, large numbers (mean approximately 4500/mm(2)) of thalamocortical neurons converge in layer 1 and that this convergence gives rise to a very high local density of thalamic terminals. Moreover, we show that the layer 1-projecting neurons are present in large numbers in most, but not all, motor, association, limbic, and sensory nuclei of the rodent thalamus. Some layer 1-projecting axons branch to innervate large swaths of the cerebral hemisphere, whereas others arborize within only a single cortical area. Present data imply that realistic modeling of cortical circuitry should factor in a dense axonal canopy carrying highly convergent thalamocortical input to pyramidal cell apical tufts. In addition, they are consistent with the notion that layer 1-projecting axons may be a robust anatomical substrate for extensive "feedback" interactions between cortical areas via the thalamus. PMID- 19188275 TI - Patterns of nocturnal rehydration in root tissues of Vaccinium corymbosum L. under severe drought conditions. AB - Although roots in dry soil layers are commonly rehydrated by internal hydraulic redistribution during the nocturnal period, patterns of tissue rehydration are poorly understood. Rates of nocturnal rehydration were examined in roots of different orders in Vaccinium corymbosum L. 'Bluecrop' (Northern highbush blueberry) grown in a split-pot system with one set of roots in relatively moist soil and the other set of roots in dry soil. Vaccinium is noted for a highly branched and extremely fine root system. It is hypothesized that nocturnal root tissue rehydration would be slow, especially in the distal root orders because of their greater hydraulic constraints (smaller vessel diameters and fewer number of vessels). Vaccinium root hydraulic properties delayed internal water movement. Even when water was readily available to roots in the wet soil and transpiration was minimal, it took a whole night-time period of 12 h for the distal finest roots (1st to 4th order) under dry soil conditions to reach the same water potentials as fine roots in moist soil (1st to 4th order). Even though roots under dry soil equilibrated with roots in moist soil, the equilibrium point reached before sunrise was about -1.2 MPa, indicating that tissues were not fully rehydrated. Using a single-branch root model, it was estimated that individual roots exhibiting the lowest water potentials in dry soil were 1st order roots (distal finest roots of the root system). However, considered at the branch level, root orders with the highest hydraulic resistances corresponded to the lowest orders of the permanent root system (3rd-, 4th-, and 5th-order roots), thus indicating possible locations of hydraulic safety control in the root system of this species. PMID- 19188276 TI - Increased sensitivity to iron deficiency in Arabidopsis thaliana overaccumulating nicotianamine. AB - Nicotianamine (NA) is a non-protein amino acid derivative synthesized from S adenosyl L-methionine able to bind several metal ions such as iron, copper, manganese, zinc, or nickel. In plants, NA appears to be involved in iron availability and is essential for the plant to complete its biological cycle. In graminaceous plants, NA is also the precursor in the biosynthesis of phytosiderophores. Arabidopsis lines accumulating 4- and 100-fold more NA than wild-type plants were used in order to evaluate the impact of such an NA overaccumulation on iron homeostasis. The expression of iron-regulated genes including the IRT1/FRO2 iron uptake system is highly induced at the transcript level under both iron-sufficient and iron-deficient conditions. Nevertheless, NA overaccumulation does not interfere with the iron uptake mechanisms since the iron levels are similar in the NA-overaccumulating line and wild-type plants in both roots and leaves under both sufficient and deficient conditions. This observation also suggests that the translocation of iron from the root to the shoot is not affected in the NA-overaccumulating line. However, NA overaccumulation triggers an enhanced sensitivity to iron starvation, associated with a decrease in iron availability. This study draws attention to a particular phenotype where NA in excess paradoxically leads to iron deficiency, probably because of an increase of the NA apoplastic pool sequestering iron. This finding strengthens the notion that extracellular NA in the apoplast could be a major checkpoint to control plant iron homeostasis. PMID- 19188277 TI - Salicylic acid deficiency in NahG transgenic lines and sid2 mutants increases seed yield in the annual plant Arabidopsis thaliana. AB - Salicylic acid-deficient NahG transgenic lines and sid2 mutants were used to evaluate the role of this compound in the development of the short-lived, annual plant Arabidopsis thaliana, with a particular focus on the interplay between salicylic acid and other phytohormones. Low salicylic acid levels led to increased growth, as well as to smaller abscisic acid levels and reduced damage to PSII (as indicated by F(v)/F(m) ratios) during the reproductive stages in rosette leaves of NahG transgenic lines and sid2 mutants, compared with wild-type plants. Furthermore, salicylic acid deficiency highly influenced seed yield and composition. Seed production increased by 4.4-fold and 3.5-fold in NahG transgenic lines and sid2 mutants, respectively, compared to the wild type. Salicylic acid deficiency also improved seed composition in terms of antioxidant vitamin concentrations, seeds of salicylic acid-deficient plants showing higher levels of alpha- and gamma-tocopherol (vitamin E) and beta-carotene (pro-vitamin A) than seeds of wild-type plants. Seeds of salicylic acid-deficient plants also showed higher nitrogen concentrations than seeds of wild-type plants. It is concluded that (i) the sid2 gene, which encodes for isochorismate synthase, plays a central role in salicylic acid biosynthesis during plant development in A. thaliana, (ii) salicylic acid plays a role in the regulation of growth, senescence, and seed production, (iii) there is a cross-talk between salicylic acid and other phytohormones during plant development, and (iv) the concentrations of antioxidant vitamins in seeds may be influenced by the endogenous levels of salicylic acid in plants. PMID- 19188278 TI - Modulation in reproductive tissue redox profile in sexually receptive female rats after short-term exposure to male chemical cues. AB - It is well known that antioxidants play an important role in sperm fertility, but there is no data on the literature regarding the effect of male chemical cues in the antioxidant defenses of the female reproductive tract. Here, we evaluated oxidative parameters in ovaries and uterus of virgin female rats isolated from contact to males and exposed only to male-soiled bedding (MSB). Four-month-old Wistar (regular 4-day cyclic) virgin female rats were utilized from proestrus to estrus phase of the reproductive cycle for experimental exposure. In an isolated room, female rats were exposed for 90 min to MSB. For biochemical assays, female rats were killed by decapitation at 30, 90, 180, and 240 min after the end of exposure, and the ovaries and uterus were removed for further analysis. Antioxidant enzyme activities (superoxide dismutase, catalase, and glutathione peroxidase), the nonenzymatic antioxidant potential (total radical-trapping antioxidant parameter), and the oxidative damage parameters (thiobarbituric acid reactive species and carbonyl content) were analyzed. We observed an increase in the nonenzymatic antioxidant potential and diminished free radical oxidative damage in uterine tissue, 30 and 90 min after exposure. Furthermore, in ovaries, enzymatic defenses were modulated distinctly along the 240 min after exposure. MSB exposure modulates the antioxidant profile in ovaries and uterus of receptive female rats. It is possible that the modifications in the oxidative profile of the female genital tract may have important implications in the process of fertilization. PMID- 19188280 TI - A candidate pheromone receptor and two odorant receptors of the hawkmoth Manduca sexta. AB - In this study, we cloned and characterized three Manduca sexta odorant receptors (ORs). One receptor is a putative pheromone receptor expressed exclusively in a cell associated with male-specific type-I trichoid sensilla. We describe the results of real-time PCR (RT-PCR) and quantitative real-time PCR (qRT-PCR) experiments that show MsextaOR1 is expressed only in male antennae. In situ hybridization labels a single cell associated with type-1 trichoid sensilla, which houses two neurons that have been previously determined to respond to the major components of the pheromone blend. The second receptor, MsextaOR2, was discovered using degenerate primers designed to conserved motifs of a unique group ORs that share as much as 88% identity. Comparison of RT-PCR, qRT-PCR, and in situ hybridization results with those of ORs in the Drosophila melanogaster Or83b subfamily shows a strong sequence and expression pattern similarity. The third receptor, MsextaOR3, was found by 5'-end sequencing of a normalized and subtracted cDNA library from male M. sexta antennae. RT-PCR and qRT-PCR show that this receptor is expressed only in male and female antennae. These are the first ORs, including a putative pheromone receptor, to be described from M. sexta. PMID- 19188279 TI - A high-throughput method to measure NaCl and acid taste thresholds in mice. AB - To develop a technique suitable for measuring NaCl taste thresholds in genetic studies, we conducted a series of experiments with outbred CD-1 mice using conditioned taste aversion (CTA) and two-bottle preference tests. In Experiment 1, we compared conditioning procedures involving either oral self-administration of LiCl or pairing NaCl intake with LiCl injections and found that thresholds were the lowest after LiCl self-administration. In Experiment 2, we compared different procedures (30-min and 48-h tests) for testing conditioned mice and found that the 48-h test is more sensitive. In Experiment 3, we examined the effects of varying strength of conditioned (NaCl or LiCl taste intensity) and unconditioned (LiCl toxicity) stimuli and concluded that 75-150 mM LiCl or its mixtures with NaCl are the optimal stimuli for conditioning by oral self administration. In Experiment 4, we examined whether this technique is applicable for measuring taste thresholds for other taste stimuli. Results of these experiments show that conditioning by oral self-administration of LiCl solutions or its mixtures with other taste stimuli followed by 48-h two-bottle tests of concentration series of a conditioned stimulus is an efficient and sensitive method to measure taste thresholds. Thresholds measured with this technique were 2 mM for NaCl and 1 mM for citric acid. This approach is suitable for simultaneous testing of large numbers of animals, which is required for genetic studies. These data demonstrate that mice, like several other species, generalize CTA from LiCl to NaCl, suggesting that they perceive taste of NaCl and LiCl as qualitatively similar, and they also can generalize CTA of a binary mixture of taste stimuli to mixture components. PMID- 19188281 TI - Morphological characterization of as-received and in vivo orthodontic stainless steel archwires. AB - This study was undertaken to evaluate the material degradation of clinical bracket-archwire-contacting surfaces after in vivo orthodontic use. Twenty-four stainless steel multiloop edgewise archwires with two different cross sections (0.016 x 0.016 and 0.016 x 0.022 inches) were used for at least 6 months in the mouths of 14 patients. The surfaces of both as-received (cross-section of 0.016 x 0.016, 0.016 x 0.022, and 0.017 x 0.025 inches) and the in vivo wires were examined using scanning electron microscopy. The as-received wires exhibited an inhomogeneous surface with different surface irregularities resulting from the manufacturing process. For the in vivo archwires, an increase in the variety, type, and number of surface irregularities were observed. Crevice corrosion occurred not only at surface irregularities formed during manufacturing and orthodontic handling but also at the bracket-archwire-contacting surfaces and at the archwire surfaces coated with plaque and food remnants. This corrosion may be linked to the formation of a micro-environment at these locations. In addition, a limited number of signs of degradation induced during in vivo testing due to wear and friction were observed. PMID- 19188284 TI - A schizophrenia gene locus on chromosome 17q21 in a new set of families of Mexican and central american ancestry: evidence from the NIMH Genetics of schizophrenia in latino populations study. AB - OBJECTIVE: The present study investigated a new set of families of Latin American ancestry in order to detect the location of genes predisposing to schizophrenia and related psychotic disorders. METHOD: A genome-wide scan was performed for 175 newly recruited families with at least two siblings suffering from a psychotic disorder. Best-estimate consensus procedures were used to arrive at diagnoses, and nonparametric allele-sharing statistics were calculated to detect linkage. RESULTS: Genome-wide significant evidence for linkage for the phenotype of DSM-IV schizophrenia or schizoaffective disorder was found in a region on chromosome 17q21 (lod score, 3.33). A region on chromosome 15q22-23 showed suggestive evidence of linkage with this same phenotype (lod score, 2.11). Analyses using a broader model (any psychosis) yielded evidence of suggestive linkage for the 17q21 region only, and no region achieved genome-wide significance of linkage. CONCLUSIONS: The new set of 175 families of Mexican and Central American ancestry delineates two new loci likely to harbor predisposition genes for schizophrenia and schizoaffective disorder. The region with the strongest support for linkage in this sample, 17q21, has been implicated in meta-analyses of schizophrenia genome screens, but the authors found no previous reports of it as a locus for schizophrenia in specific population- or family-based studies, and it may represent the location of a schizophrenia predisposition gene (or genes) of special relevance in Mexican and Central American populations. PMID- 19188283 TI - Obsessions and compulsions in the community: prevalence, interference, help seeking, developmental stability, and co-occurring psychiatric conditions. AB - OBJECTIVE: It is unclear how many people in the community have obsessions and compulsions and associated levels of interference. It is also unknown what variables predict help-seeking for these symptoms, whether they are developmentally stable, and whether they increase the risk of mental disorders. METHOD: The authors analyzed data from the prospective longitudinal Dunedin study of an unselected birth cohort. The presence of obsessions and compulsions and mental disorders was assessed using the Diagnostic Interview Schedule (DIS) at ages 11, 26, and 32. Data on interference and help-seeking were obtained at ages 26 and 32. RESULTS: Obsessions and compulsions were frequent in individuals with mental disorders other than obsessive-compulsive disorder (OCD) and among people without mental disorders. Even in the latter group, these symptoms caused significant interference. The presence of anxiety/depression and of obsessions (particularly aggressive and shameful thoughts), but not compulsions, was associated with help-seeking. Harm/checking was the most prevalent symptom dimension. Symptom dimensions were temporally stable and associated with increased comorbidity. Obsessive-compulsive symptoms at age 11 predicted a high risk of an adult OCD diagnosis as well as elevated adult symptom dimensions. CONCLUSIONS: Obsessions and compulsions are common in the adult population, have their roots in childhood, and are associated with interference, risk for disorders, and help-seeking. Subclinical obsessions and compulsions should be taken into account in research, intervention, and DSM-V. PMID- 19188285 TI - Systematic review and meta-analysis of multiple-session early interventions following traumatic events. AB - OBJECTIVE: The authors sought to determine the efficacy of multiple-session psychological interventions to prevent and treat traumatic stress symptoms beginning within 3 months of a traumatic event. METHOD: Nine computerized databases were searched, and manual searches were conducted of reference lists of selected articles as well as two journals. In addition, key researchers in the field were contacted to determine whether they were aware of other relevant studies. The reviewers identified randomized controlled trials of multiple session psychological treatments aimed at preventing or reducing traumatic stress symptoms in individuals within 3 months of exposure to a traumatic event. Details of the studies were independently extracted by two reviewers, and outcome data were entered into the Review Manager software package. Quality assessment was also conducted by two researchers independently. RESULTS: Twenty-five studies examining a range of interventions were identified. For treatment of individuals exposed to a trauma irrespective of their symptoms, there was no significant difference between any intervention and usual care. For treatment of traumatic stress symptoms irrespective of diagnosis, trauma-focused cognitive-behavioral therapy (CBT) was more effective than waiting list or supportive counseling conditions. The difference was greatest for treatment of acute stress disorder and acute posttraumatic stress disorder. CONCLUSIONS: Trauma-focused CBT within 3 months of a traumatic event appears to be effective for individuals with traumatic stress symptoms, especially those who meet the threshold for a clinical diagnosis. PMID- 19188286 TI - Mixed depression: the importance of rediscovering subtypes of mixed mood States. PMID- 19188287 TI - Toward revising the ethical boundaries of research with noncompetent subjects. PMID- 19188288 TI - Connecting brain structure and function in schizophrenia. PMID- 19188289 TI - Peripheral olfactory system for clinical and basic psychiatry: a promising entry point to the mystery of brain mechanism and biomarker identification in schizophrenia. PMID- 19188290 TI - Publication bias and the efficacy of antidepressants. PMID- 19188291 TI - Psychosis in the elderly. PMID- 19188292 TI - Oregon State Hospital, 1883-2008. PMID- 19188293 TI - Examining housing policy for persons with severe mental illness. PMID- 19188294 TI - Housing first for those with chronic mental illness. PMID- 19188295 TI - Asperger's syndrome and Autistic disorder: clearly differentiating the diagnostic criteria. PMID- 19188296 TI - Add-on filgrastim during clozapine rechallenge in patients with a history of clozapine-related granulocytopenia/agranulocytosis. PMID- 19188297 TI - Memantine as an augmenting agent for severe pediatric OCD. PMID- 19188298 TI - In brief: adherence. PMID- 19188299 TI - Pulmonary hypertension. PMID- 19188300 TI - Adolescent immunizations. PMID- 19188301 TI - Cultural competence in clinician communication. PMID- 19188302 TI - Index of suspicion. PMID- 19188303 TI - Focus on diagnosis: the erythrocyte sedimentation rate and the C-reactive protein test. PMID- 19188304 TI - Complementary, holistic, and integrative medicine: fever. PMID- 19188306 TI - Forget the diffusion--do we need T2-weighted MR images to detect early central nervous system injury? AB - SUMMARY: T2- and diffusion-weighted (DW) magnetic resonance (MR) imaging examinations performed with techniques adopted from the brain are practical in the spinal cord. Results indicate that these modalities will be highly useful for characterizing radiation injury to the spinal cord and illuminate our understanding of the brain's response to radiation therapy. Further technical developments will be required to make high-spatial-resolution, high-quality axial DW imaging practical in routine clinical use. PMID- 19188307 TI - What is the current role of CT urography and MR urography in the evaluation of the urinary tract? AB - Technologic advances in both computed tomography (CT) and magnetic resonance (MR) imaging have resulted in the ability to image the urinary tract in ways that surpass the prior mainstay of urinary tract imaging, the intravenous urogram. In adults, for most, if not all, historical indications for intravenous urography, CT urography or MR urography is now the preferred examination. Although a variety of techniques for both examinations have been described, each test provides more diagnostic information than does intravenous urography. With the introduction of multidetector technology, CT urography, to date, has emerged as the initial heir apparent to intravenous urography; many years of experience have now clearly demonstrated that CT is the test of choice for many urologic problems, including urolithiasis, renal masses, urinary tract infection, trauma, and obstructive uropathy. CT urography provides a detailed anatomic depiction of each of the major portions of the urinary tract--the kidneys, intrarenal collecting systems, ureters, and bladder--and thus allows patients with hematuria to be evaluated comprehensively. MR urography can be used also to evaluate the urinary tract and has the advantage of not using ionizing radiation and the potential to provide more functional information than CT. However, MR urography is less established and less reliably results in diagnostic image quality relative to CT urography. Although both tests can be used to evaluate the urinary tract, several issues remain and include reaching a consensus on the optimal protocols and appropriate utilization in an era of cost containment and heightened concerns about radiation exposure. PMID- 19188308 TI - Unresectable hepatocellular carcinoma: serial early vascular and cellular changes after transarterial chemoembolization. PMID- 19188309 TI - CT venography: a necessary adjunct to CT pulmonary angiography or a waste of time, money, and radiation? PMID- 19188310 TI - Motion in cardiovascular MR imaging. AB - Modern rapid magnetic resonance (MR) imaging techniques have led to widespread use of the modality in cardiac imaging. Despite this progress, many MR studies suffer from image degradation due to involuntary motion during the acquisition. This review describes the type and extent of the motion of the heart due to the cardiac and respiratory cycles, which create image artifacts. Methods of eliminating or reducing the problems caused by the cardiac cycle are discussed, including electrocardiogram gating, subject-specific acquisition windows, and section tracking. Similarly, for respiratory motion of the heart, techniques such as breath holding, respiratory gating, section tracking, phase-encoding ordering, subject-specific translational models, and a range of new techniques are considered. PMID- 19188311 TI - Consensus review of discordant findings maximizes cancer detection rate in double reader screening mammography: Irish National Breast Screening Program experience. AB - PURPOSE: To assesses consensus review of discordant screening mammography findings in terms of its sensitivity, safety, and effect on overall performance in the first 6 years of operation of the Irish National Breast Screening Program (NBSP). MATERIALS AND METHODS: Women who participated in the Irish NBSP gave written informed consent for use of their data for auditing purposes. Local ethics committee approval was obtained. The study population consisted of women who participated in the Irish NBSP and underwent initial screening mammography at one of the two screening centers serving the eastern part of Ireland between 2000 and 2005. Independent double reading of mammograms was performed. When the readers disagreed regarding referral, the case was reviewed by a consensus panel. Of the 128 569 screenings performed, 1335 (1%) were discussed by consensus. RESULTS: Of the 1335 cases discussed by consensus, 606 (45.39%) were recalled for further assessment. This resulted in an overall recall rate of 4.41%. In those recalled to assessment, 71 cases of malignant disease were diagnosed (ductal carcinoma in situ, n = 24; invasive cancer, n = 47). The remaining 729 patients were returned to biennial screening. Of these 729 patients, seven had false negative findings that were identified in the subsequent screening round. Use of the highest reader recall method, in which a patient is recalled if her findings are deemed abnormal by either reader, could potentially increase the cancer detection rate by 0.6 per 1000 women screened but would increase the recall rate by 12.69% and the number of false-positive findings by 15.37%. CONCLUSION: The consensus panel identified 71 (7.33%) of 968 cancers diagnosed. Consensus review substantially reduced the number of cases recalled and was associated with a low false-negative rate. PMID- 19188312 TI - Nephrogenic systemic fibrosis: incidence, associations, and effect of risk factor assessment--report of 33 cases. AB - PURPOSE: To describe the presentation and clinical course of patients with nephrogenic systemic fibrosis (NSF) at a large acute-care hospital, to evaluate the overall incidence of NSF, and to assess the effect of a hospital-wide policy regarding gadolinium-based contrast agent (GBCA) use on NSF incidence. MATERIALS AND METHODS: A review of all cases of NSF observed at an institution from 2003 to 2008 was conducted. This HIPAA-compliant study was approved by the institutional review board. The informed consent requirement was waived. Demographics, medical history, and associated conditions were recorded. Radiologic procedures were evaluated if they were performed within 1 year prior to NSF onset. GBCA use was assessed by checking the electronic database for each procedure. The incidence of NSF was compared before and after implementation of an institutional policy designed to assess risk of NSF prior to GBCA use. RESULTS: All 33 patients with NSF (mean age, 49 years; age range, 15-78 years) had advanced renal failure (estimated glomerular filtration rate < 15 mL/min/1.73 m(2)) when the GBCA was injected. Twenty-six patients had severe chronic or end-stage renal disease, and seven had acute renal failure. The mean interval between contrast material injection and NSF onset was 29 days +/- 25 (standard deviation) (range, 4-112 days). The overall incidence of NSF was 36.5 cases per 100,000 gadolinium enhanced magnetic resonance (MR) examinations between 2003 and 2006 and four cases per 100,000 gadolinium-enhanced MR examinations between 2007 and 2008 after screening for NSF risk was instituted (Fisher exact test, P = .001). Five patients developed NSF in the peritransplant period, and four underwent a catheter-based radiographic procedure with administration of a GBCA. CONCLUSION: Common associations of GBCA MR imaging and NSF were acute and severe chronic renal failure and liver or renal transplantation. Screening procedures performed before MR imaging to determine which patients were at risk of developing NSF appear to reduce the incidence of this complication and further support the belief that NSF is associated with GBCA administration. PMID- 19188313 TI - Imaging nanoprobe for prediction of outcome of nanoparticle chemotherapy by using mammography. AB - PURPOSE: To prospectively predict the effectiveness of a clinically used nanochemotherapeutic agent by detecting and measuring the intratumoral uptake of an x-ray contrast agent nanoprobe by using digital mammography. MATERIALS AND METHODS: All animal procedures were approved by the institutional animal care and use committee. A long-circulating 100-nm-scale injectable liposomal probe encapsulating 155 mg/mL iodine was developed. Preliminary studies were performed to identify the agent dose that would result in adequate tumor enhancement without enhancement of the normal vasculature in rats. This dose was used to image a rat breast tumor (n = 14) intermittently for 3 days by using a digital mammography system; subsequently, the animals were treated with liposomal doxorubicin. The predictive capability of the probe was characterized by creating good- and bad-prognosis subgroups, on the basis of tumor enhancement found during imaging, and analyzing the tumor growth after treatment of the animals in these two subgroups. RESULTS: A dose of 455 mg of iodine per kilogram of body weight was found to produce an undetectable signal from the blood while achieving enough intratumoral accumulation of the probe to produce adequate signal for detection. The good- and bad-prognosis subgroups demonstrated differential tumor growth rates (P < .003). An inverse linear relationship between the contrast enhancement rate constant during imaging and the tumor growth rate constant during treatment was found (slope = -0.576, R(2) = 0.838). CONCLUSION: In this animal model, quantitative measurement of vascular permeability enabled prediction of therapeutic responsiveness of tumors to liposomal doxorubicin. PMID- 19188314 TI - Quantitative CT color mapping of the arterial enhancement fraction of the liver to detect hepatocellular carcinoma. AB - PURPOSE: To investigate the diagnostic value of quantitative determination and three-dimensional color mapping of the arterial enhancement fraction (AEF) of the liver for the detection of hepatocellular carcinoma (HCC). MATERIALS AND METHODS: This retrospective study was approved by the institutional review board; informed consent was waived. Eighty-two patients (mean age, 56.2 years +/- 10.7 [standard deviation]), 69 men (mean age, 57.1 years +/- 10.0) and 13 women (mean age, 51.6 years +/- 13.6), were followed-up for HCC or liver cirrhosis by using available multiphasic multidetector computed tomographic (CT) scans. Quantitative AEF color maps (the ratio of the attenuation increment during the arterial phase to the attenuation increment during the portal venous phase) of the whole liver were created from routine CT images by using prototypic software. Three radiologists independently analyzed a multiphasic CT image set; 4 weeks later, to avoid any potential recall bias, they independently analyzed a combined image set of the quantitative color maps and the multiphasic CT images. The additional diagnostic value of the color mapping was evaluated by means of jackknife alternative free response receiver-operating characteristic (JAFROC) analysis. RESULTS: The mean AEF of HCCs (75.6 +/- 13.3) was significantly higher than that of liver parenchyma (23.4 +/- 8.2) (P < .001). The average JAFROC figure of merit was 0.861 on the initial multiphasic CT image set. This increased to 0.933 on the second session with the combined image set (P < .05). The mean sensitivity for HCC detection increased from 71.7% with the multiphasic CT image set to 88.8% with the combined image set. CONCLUSION: Quantitative color mapping of the AEF can increase the sensitivity and diagnostic performance of multiphasic multidetector CT for detecting HCC. Supplementamaterial:http://radiology.rsnajnls.org/cgi/content/full/250/2/425/DC1h tp://radiology.rsnajnls.org/cgi/content/full/250/2/425/DC2http://radiology.rsnajn s.org/cgi/content/full/250/2/425/DC3 PMID- 19188316 TI - Comparison of four embolic materials at uterine artery embolization by using postprocedural MR imaging enhancement. AB - PURPOSE: To test the hypothesis that not all embolic materials are equivalent by using postprocedural magnetic resonance (MR) imaging enhancement of uterine fibroids in patients treated with uterine artery embolization (UAE). MATERIALS AND METHODS: Approval and a waiver of consent from the institutional human investigations committee was received for this study. The study was HIPAA compliant. A total of 84 women who underwent 6-month MR imaging follow-up constituted this retrospective study. Within this group, 25 women were treated with Contour polyvinyl alcohol (PVA) particles, 23 were treated with Contour SE particles, 19 were treated with Embosphere microspheres, and 17 were treated with Bead Block microspheres. Pre- and postprocedural MR imaging results were analyzed for the total number of fibroids present in the uterus of each patient and for the percentage of individual fibroid enhancement. Enhancement of individual fibroids was measured with quartile intervals. Greater than 25% residual enhancement of a fibroid after embolization was considered an incomplete infarction. The overall percentage change in enhancement was calculated for each patient. Bivariate analysis by using generalized linear modeling and one-way analysis of variance was used to assess differences in infarction with different embolic materials. RESULTS: Among patients treated with Contour and Embosphere, there was a mean reduction in enhancement by 76.60% and 83.07%, respectively, compared with a mean reduction of 52.53% and 49.78% in patients treated with Bead Block and Contour SE, respectively. There was a significant difference in postembolization enhancement between Bead Block and Embosphere, Bead Block and Contour, Contour SE and Embosphere, and Contour SE and Contour. CONCLUSION: Patients treated with Bead Block or Contour SE demonstrated a reduced degree of infarction at follow-up MR imaging compared with patients treated with Contour or Embosphere. PMID- 19188315 TI - Unresectable hepatocellular carcinoma: serial early vascular and cellular changes after transarterial chemoembolization as detected with MR imaging. AB - PURPOSE: To prospectively assess serial changes in contrast material-enhanced and diffusion-weighted (DW) magnetic resonance (MR) imaging values within 1 month after transarterial chemoembolization (TACE) in patients with unresectable hepatocellular carcinoma (HCC). MATERIALS AND METHODS: Institutional review board approval was obtained for this prospective HIPAA-compliant study. MR imaging was performed before and within 24 hours after TACE in 24 patients with HCC (21 male, three female; mean age, 59 years and 62 years, respectively). Serial MR imaging was subsequently performed 1, 2, 3, and 4 weeks after therapy. The imaging protocol included fast spin-echo T2-weighted MR imaging, breath-hold DW echo planar MR imaging, and breath-hold unenhanced and contrast-enhanced T1-weighted three-dimensional fat-suppressed gradient-recalled-echo MR imaging in the arterial and portal venous phases. Tumor size, enhancement, and apparent diffusion coefficient (ADC) values were recorded before and sequentially after treatment. Regression models for the correlated data were used to assess changes in these parameters over time after TACE. RESULTS: Mean tumor size was 7.5 cm and was unchanged up to 4 weeks after therapy. Reduction in tumor enhancement in the arterial phase occurred immediately after TACE, with a consistent reduction occurring 1-3 weeks after therapy (P = .001). Reduction in tumor enhancement in the portal venous phase also occurred immediately after TACE, with a consistent reduction occurring 1-3 weeks after therapy (P = .0003). The increase in tumor ADC value was significant 1-2 weeks after therapy (P = .004), borderline significant 3 weeks after therapy, and insignificant 24 hours and 4 weeks after therapy. CONCLUSION: Significant reduction in tumor enhancement occurred within 24 hours after TACE and persisted up to 4 weeks after TACE. Lesser changes in the ADC value appeared 1 week after TACE, persisted through 2 weeks after TACE, and became less apparent 3 and 4 weeks after TACE. No change in tumor size was recorded during the follow-up period. PMID- 19188317 TI - Impact of computer-aided detection on the cost-effectiveness of CT colonography. AB - PURPOSE: To analyze the cost-effectiveness of adding computer-aided detection (CAD) to a computed tomographic (CT) colonography screening program and to compare it with other options of colorectal cancer (CRC) prevention. MATERIALS AND METHODS: The cost-effectiveness of screening strategies by using CT colonography with and without CAD, flexible sigmoidoscopy (FS), and optical colonoscopy were compared by using a Markov-based computer model. In the model, a hypothetical population of 100,000 persons aged 50 years underwent colorectal screening every 10 years. Baseline sensitivities for both experienced and inexperienced readers and the incremental accuracy when adding CAD were estimated from a systematic review of the literature. RESULTS: At baseline, the addition of CAD resulted in 9% and 2% increases in CRC prevention rates for inexperienced and experienced readers, respectively, when compared with CT colonography without CAD. Assuming a CAD cost of $50 per CT colonography, the overall program costs increased by only 3%-5%, largely because of the substantial reduction in CRC related costs. The incremental cost-effectiveness of CT colonography with CAD compared with CT colonography without CAD was $8661 and $61,354 per life-year gained for inexperienced and experienced readers, respectively. Optical colonoscopy was not a cost-effective alternative to CT colonography with CAD performed by experienced readers, with an incremental cost-effectiveness of $498,668 per life-year gained. CT colonography with CAD for inexperienced readers was more clinically effective and cost-effective than FS. At analysis, sensitivity of CT colonography with CAD for polyps 6 mm or larger was the most meaningful variable. CONCLUSION: The addition of CAD to CT colonography screening improves the CRC prevention rate, resulting in advantageous cost-effectiveness for screening. SUPPLEMENTAL MATERIAL: http://radiology.rsnajnls.org/cgi/content/full/250/2/488/DC1. PMID- 19188318 TI - Three-dimensional isotropic shoulder MR arthrography: comparison with two dimensional MR arthrography for the diagnosis of labral lesions at 3.0 T. AB - PURPOSE: To compare the diagnostic accuracy of three-dimensional (3D) isotropic magnetic resonance (MR) arthrography with two-dimensional (2D) MR arthrography for the diagnosis of labral lesions of the shoulder performed by using a 3.0-T imager. MATERIALS AND METHODS: Institutional review board approval was given for this retrospective study and informed consent was waived. From March 2006 to January 2007, 100 patients underwent 3D isotropic and 2D MR shoulder arthrographic imaging by using a 3.0-T imager and subsequent arthroscopic surgery. The 3D isotropic sequences were performed by using fast gradient-echo imaging with fat suppression (voxel size, 0.6 x 0.6 x 0.6 mm; imaging time, 5 minutes 32 seconds) and were evaluated for the presence of superior, anterior, and posterior labral lesions by using 3D isotropic and 2D MR arthrography. The statistical differences between the sensitivity and specificity for both methods were analyzed by using the McNemar test, with arthroscopic findings regarded as reference standard. RESULTS: Surgical findings confirmed 53 superior labral anterior posterior (SLAP) lesions, 17 anterior labral lesions, and five posterior labral lesions. Respective sensitivity and specificity were 85% and 96% for SLAP lesions, 100% and 98% for anterior labral lesions, and 80% and 99% for posterior labral lesions by using 2D MR arthrography and 83% and 96% for SLAP lesions, 100% and 96% for anterior labral lesions, and 80% and 99% for posterior labral lesions by using 3D isotropic MR arthrography. There was no significant difference in sensitivities and specificities of both methods; however, the power of this study was limited and larger comparison is needed. CONCLUSION: Isotropic 3D shoulder MR arthrography combined with a multiplanar reconstruction technique can help in the diagnosis of shoulder labral lesions as does 2D MR arthrography, but with shorter imaging times. PMID- 19188319 TI - Adrenal nodules at FDG PET/CT in patients known to have or suspected of having lung cancer: a proposal for an efficient diagnostic algorithm. AB - PURPOSE: To develop an algorithm to maximize the diagnostic yield of positron emission tomography (PET)/computed tomography (CT) by using defined attenuation and standardized uptake value (SUV) criteria. MATERIALS AND METHODS: An IRB approved, HIPAA-compliant retrospective review with waiver of informed consent of data in 1388 consecutive patients who underwent PET/CT for known or suspected lung cancer was completed, and 187 adrenal nodules were identified in 147 patients. Nodules were defined histologically or by size change (malignant, n = 37) or stability for more than 1 year (benign, n = 58). Nodules not sampled for biopsy and with less than 1 year of follow-up were considered indeterminate (n = 92). Diameter, mean attenuation, SUV(max), and SUV ratio (nodule SUV(max)/liver SUV(avg)) were compared with t test and receiver operating characteristic analyses. Sensitivity, specificity, positive predictive value, and negative predictive value were calculated for diameter > 3 cm, mean attenuation > 10 HU, nodule SUV(max) > 3.1, and SUV ratio > 1.0. These were also calculated for higher SUV(max) and SUV ratio thresholds that were found to exclude all false-positives. Diagnostic accuracy was compared by using the McNemar test (P < .05). RESULTS: In the study group of 147 patients (aged 42-88 years; mean, 65.5 years; 59 women), combined PET/CT with mean attenuation > 10 HU and SUV(max) > 3.1 had 97.3% sensitivity and 86.2% specificity. Combined PET/CT with mean attenuation > 10 HU and SUV ratio > 1.0 had 97.3% sensitivity and 74.1% specificity. The accuracies of these threshold combinations (90.5% and 83.2%, respectively) were significantly different (P = .008). Applying a further cutoff of SUV ratio > 2.5 enabled identification of 22 of 37 metastatic lesions and exclusion of all fluorodeoxyglucose-avid benign nodules. CONCLUSION: Definitive identification of many metastases can be accomplished by applying an SUV ratio cutoff of greater than 2.5, allowing pragmatic management of adrenal nodules that initially test positive with the combined PET/CT criteria SUV(max) > 3.1 and mean attenuation > 10 HU. SUPPLEMENTAL MATERIAL: http://radiology.rsnajnls.org/cgi/content/full/250/2/523/DC1. PMID- 19188321 TI - Occupational health hazards in the interventional laboratory: time for a safer environment. AB - This document is a consensus statement by the major American societies of physicians who work in the interventional laboratory environment. It reviews available data on the prevalence of occupational health risks and summarizes ongoing epidemiologic studies designed to further elucidate these risks. Its purpose is to affirm that the interventional laboratory poses workplace hazards that must be acknowledged, better understood, and mitigated to the greatest extent possible. Vigorous efforts are advocated to reduce these hazards. Interventional physicians and their professional societies, working together with industry, should strive toward minimizing operator radiation exposure, eliminating the need for personal protective apparel, and ending the orthopedic and ergonomic consequences of the interventional laboratory work environment. PMID- 19188320 TI - Suspected appendicitis in children: diagnostic importance of normal abdominopelvic CT findings with nonvisualized appendix. AB - PURPOSE: To determine whether lack of visualization of the appendix on otherwise normal abdominopelvic computed tomographic (CT) images can help exclude appendicitis in the pediatric population. MATERIALS AND METHODS: The study was institutional review board approved and HIPAA compliant. One thousand one hundred thirty-nine children suspected of having appendicitis were referred for CT examination between July 2002 and December 2006. Exclusion criteria included CT diagnosis of appendicitis or other cause of symptoms and lack of clinical follow up. Consensus review was performed by two pediatric radiologists to determine normal examinations, leaving a final study group (nonvisualized appendix) of 156 patients (mean age, 9.6 years; boys, 7.2 years; girls, 10.2 years) and a control group (visualized appendix) of 421 patients (mean age, 11.0 years; boys, 9.8 years; girls, 11.2 years). In the control group, there were 168 subjects with a partially visualized (PV) appendix and 253 with a fully visualized (FV) appendix. Pericecal fat was graded according to published criteria. Diagnosis was confirmed at surgery or clinical follow-up. Negative predictive values were calculated with 95% confidence intervals (CIs). RESULTS: There were three false-negative findings (study group, two; control group, one [FV]). The negative predictive value of a normal CT examination in pediatric patients with a nonvisualized appendix was 98.7% (95% CI: 95.5%, 99.8%); that with a visualized appendix, 99.8% (95% CI: 98.7%, 99.99%); that with a PV appendix, 100% (95% CI: 97.8%, 100%); and that with a FV appendix, 99.6% (95% CI: 97.8%, 99.99%). CONCLUSION: Pediatric abdominopelvic CT images with nonvisualized appendix have a high negative predictive value, without significant difference from cases with a PV or even FV appendix. The false-negative rate was similar to those reported in two adult series. PMID- 19188322 TI - Effective dose during abdominal three-dimensional imaging with a flat-panel detector angiography system. AB - The purpose of this study was to measure the effective dose during abdominal three-dimensional imaging obtained with an angiography unit with a digital flat panel system on a phantom and to determine dose-area product (DAP)-to-effective dose conversion factors. DAPs and effective doses were evaluated for 163-cm-tall human-shaped phantoms with estimated body weights of 54, 64, and 77 kg, and the effective doses were 2.1, 3.2, and 4.2 mSv, respectively. The DAP-to-effective dose conversion factors were 0.28-0.29 mSv x Gy(-1) x cm(-2). In conclusion, the DAPs were useful for estimating the effective dose during abdominal three dimensional angiographic imaging. PMID- 19188323 TI - Enhancement of visual perception with use of dynamic cues. AB - Institutional review board approval and signed informed consent were not needed, as medical images included in public databases were used in this study. The purpose of this study was to improve the detection of microcalcifications on mammograms and lung nodules on chest radiographs by using the dynamic cues algorithm and the motion and flickering sensitivity of the human visual system (HVS). Different sets of mammograms from the Mammographic Image Analysis Society database and chest radiographs from the Japanese Society of Radiological Technology database were presented statically, as is standard, and in a video sequence generated with the dynamic cues algorithm. Nine observers were asked to rate the presence of abnormalities with a five-point scale (1, definitely not present; 5, definitely present). The data were analyzed with receiver operating characteristic (ROC) techniques and the Dorfman-Berbaum-Metz method. The video sequence generated with the dynamic cues algorithm increased the rate of detection of microcalcifications by 10.2% (P = .002) compared with that obtained with the standard static method, as measured by the area under the ROC curve. Similar results were obtained for lung nodules, with an increase of 12.3% (P = .0054). The increase in the rate of correct detection did not come just from the image contrast change produced by the algorithm but also from the fact that image frames generated with the dynamic cues algorithm were put together in a video sequence so that the motion sensitivity of the HVS could be used to facilitate the detection of low-contrast objects. SUPPLEMENTAL MATERIAL: http://radiology.rsnajnls.org/cgi/content/full/250/2/551/DC1. PMID- 19188324 TI - Pulmonary hypertension: ECG-gated 64-section CT angiographic evaluation of new functional parameters as diagnostic criteria. AB - PURPOSE: To evaluate, by using electrocardiographically (ECG)-gated multidetector computed tomography (CT), parameters such as right ventricular outflow tract (RVOT) systolic shortening and myocardial thickness and right pulmonary artery (RPA) distensibility, reported to be abnormal at echocardiography and/or magnetic resonance (MR) imaging in patients with pulmonary hypertension (PHT). MATERIALS AND METHODS: The institutional review board approved the study and waived informed consent. Forty-five patients, 21 with PHT (group 1; mean pulmonary arterial pressure [PAP], 42 mm Hg) and 24 without (group 2; mean PAP, 16.5 mm Hg), who had undergone thoracic ECG-gated 64-section CT and right heart catheterization as part of their diagnostic work-up were included in this study. Two independent observers measured RPA distensibility and RVOT myocardial thickness, diameter, and cross-sectional area during systole and diastole. Their values were compared in both groups (Mann-Whitney U test). The area under the receiver operating characteristic (ROC) curve and Spearman correlation with mean PAP were also obtained. RESULTS: Interobserver agreement was good for all measurements (R > 0.8) except for systolic RVOT wall thickness. Median values of RPA distensibility, diastolic RVOT wall thickness, and systolic RVOT diameter and cross-sectional area were significantly different between groups 1 and 2. The largest area under the ROC curve was obtained with RPA distensibility (0.951; 95% confidence interval: 0.89, 1) also showing the closest correlation with mean PAP (r = -0.79; P < .0001). CONCLUSION: Among all parameters evaluated, RPA distensibility shows the best diagnostic value for PHT and could be useful for risk stratification. PMID- 19188325 TI - Changes in regional airflow obstruction over time in the lungs of patients with asthma: evaluation with 3He MR imaging. AB - PURPOSE: To determine changes in regional airflow obstruction over time in the lungs of patients with asthma, as demonstrated with hyperpolarized helium 3 ((3)He) magnetic resonance (MR) imaging, and to assess correlations with disease severity and use of asthma medications. MATERIALS AND METHODS: Institutional review board approval and written informed consent were obtained for this HIPAA compliant study. Use of (3)He was approved by the U.S. Food and Drug Administration. Forty-three patients underwent 103 MR imaging studies in total; 26 were imaged twice within 42-82 minutes (same day), and 17 were imaged on 3 days between 1 and 476 days (multiday). Each day, spirometry was performed, disease severity was determined, and the use of asthma medications was recorded. Images were reviewed in a pairwise fashion to determine total ventilation defect number, defects in same location between imaging studies, and size. Parametric and nonparametric statistical methods were used. RESULTS: For the same-day examinations, the mean number of defects per image section was similar at baseline and repeat imaging (1.8 +/- 1.9 [standard deviation] vs 1.6 +/- 1.9, respectively; P = .15), with 75% of defects remaining in the same location and 71% of these not changing size. For the multiday examinations, the mean number of defects per section was higher for study 2 (2.4 +/- 1.5) than study 1 (1.7 +/- 0.9, P = .02), was lower for study 3 (1.5 +/- 1.1) than for study 2 (P < .01), and was similar for studies 1 and 3 (P = .56). Time between examinations was not associated with change in mean number of defects per section (median intrasubject correlation [r(m)] = 0.01, P = .64) or change in spirometric values (range of r(m) values: -0.56 to -0.31; range of P values: .09-.71). Defects in the same location decreased with time (r(m) = -0.83, P < .01), with 67% persisting between studies 1 and 2 (median interval, 31 days), 43% persisting between studies 2 and 3 (median interval, 41 days), and 38% persisting between studies 1 and 3 (median interval, 85 days); 46%-58% of defects remained unchanged in size. These trends were the same regardless of disease severity or medication use. CONCLUSION: In asthma, focal airflow impediment within the lungs can be markedly persistent over time, regardless of disease severity or treatment. PMID- 19188326 TI - Lung nodules: CT-guided placement of microcoils to direct video-assisted thoracoscopic surgical resection. AB - PURPOSE: To prospectively assess the safety and effectiveness of computed tomography (CT)-guided placement of fiber-coated microcoils used to guide video assisted thoracoscopic surgical (VATS) excision of small peripheral lung nodules, with successful excision as the primary outcome and successful CT-guided microcoil placement and procedural complications as secondary outcomes. MATERIALS AND METHODS: The institutional review board approved the study protocol. Informed consent was obtained from all 69 enrolled patients (30 men, 39 women; mean age, 60.7 years +/- 10.1 [standard deviation]) with 75 nodules. At CT, one end of an 80-mm long, 0.018-inch-diameter fiber-coated microcoil was placed deep to the small peripheral lung nodule, and the other end was coiled in the pleural space. VATS excision of lung tissue, nodules, and the microcoil was performed with fluoroscopic guidance. RESULTS: Seventy-three (97%) 4-24-mm nodules were successfully removed at fluoroscopically guided VATS excision; two nodules could not be removed. CT-guided microcoil placement was successful in all cases; however, two (3%) of 75 coils were displaced at VATS excision. Pneumothorax requiring chest tube placement occurred in two (3%) patients, and asymptomatic hemothorax occurred in one (1%) patient. The microcoil did not impede intraoperative frozen-section histopathologic analysis, which facilitated accurate clinical management in all patients. For 19 (28%) patients, the preoperative treatment plan based on bronchoscopy, needle biopsy, and positron emission tomography findings changed after VATS excision. CONCLUSION: Microcoil localization of small peripheral lung nodules enabled fluoroscopically guided VATS resection of 97% of the nodules, with a low rate of intervention (3%) for procedural complications. PMID- 19188327 TI - Intermittent claudication: clinical effectiveness of endovascular revascularization versus supervised hospital-based exercise training--randomized controlled trial. AB - PURPOSE: To compare clinical success, functional capacity, and quality of life during 12 months after revascularization or supervised exercise training in patients with intermittent claudication. MATERIALS AND METHODS: This study had institutional review board approval, and all patients gave written informed consent. Between September 2002 and September 2005, 151 consecutive patients who presented with symptoms of intermittent claudication were randomly assigned to undergo either endovascular revascularization (angioplasty-first approach) (n = 76) or hospital-based supervised exercise (n = 75). The outcome measures were clinical success, functional capacity, and quality of life after 6 and 12 months. Clinical success was defined as improvement in at least one category in the Rutherford scale above the pretreatment level. Significance of differences between the groups was assessed with the unpaired t test, chi(2) test, or Mann Whitney U test. To adjust outcomes for imbalances of baseline values, multivariable regression analysis was performed. RESULTS: Immediately after the start of treatment, patients who underwent revascularization improved more than patients who performed exercise in terms of clinical success (adjusted odds ratio [OR], 39; 99% confidence interval [CI]: 11, 131; P < .001), but this advantage was lost after 6 (adjusted OR, 0.9; 99% CI: 0.3, 2.3; P = .70) and 12 (adjusted OR, 1.1; 99% CI: 0.5, 2.8; P = .73) months. After revascularization, fewer patients showed signs of ipsilateral symptoms at 6 months compared with patients in the exercise group (adjusted OR, 0.4; 99% CI: 0.2, 0.9; P < .001), but no significant differences were demonstrated at 12 months. After both treatments, functional capacity and quality of life scores increased after 6 and 12 months, but no significant differences between the groups were demonstrated. CONCLUSION: After 6 and 12 months, patients with intermittent claudication benefited equally from either endovascular revascularization or supervised exercise. Improvement was, however, more immediate after revascularization. PMID- 19188329 TI - Case 142: Susac syndrome. PMID- 19188330 TI - Radiologists are physicians, not commodities. PMID- 19188331 TI - Effective dose and dose-length product in CT. PMID- 19188332 TI - Radiologists as physicians. PMID- 19188333 TI - A new era for sedation in ICU patients. PMID- 19188335 TI - NHERF1 regulates parathyroid hormone receptor desensitization: interference with beta-arrestin binding. AB - Type 1 parathyroid hormone receptor (PTH1R) activation, desensitization, internalization, and recycling proceed in a cyclical manner. The Na(+)/H(+) exchange regulatory factor 1 (NHERF1) is a cytoplasmic adapter protein that regulates trafficking and signaling of several G protein-coupled receptors (GPCRs) including the PTH1R. The mineral ion wasting and bone phenotype of NHERF1 null mice suggests that PTH1R may interact with NHERF1. The objective of this study was to examine the effect of NHERF1 on PTH1R desensitization. Using rat osteosarcoma T6-N4 cells expressing the endogenous PTH1R, in which NHERF1 expression could be induced by tetracycline, PTH1R desensitization was assessed by measuring adenylyl cyclase activity after successive PTH challenges. PTH1R mediated adenylyl cyclase responses were desensitized by repetitive PTH challenges in a concentration-dependent manner, and desensitization was inhibited by NHERF1. NHERF1 blocked PTH-induced dissociation of the PTH1R from Galpha(s). Blocking PTH1R endocytosis did not mitigate PTH1R desensitization. Reducing constitutive NHERF1 levels in human osteosarcoma SAOS2 cells, which express both endogenous PTH1R and NHERF1, with short hairpin RNA directed against NHERF1 restored PTH1R desensitization. Mutagenesis of the PDZ-binding domains or deletion of the NHERF1 MERM domain demonstrated that both are required for inhibition of receptor desensitization. A phosphorylation-deficient PTH1R exhibited reduced desensitization and interaction with beta-arrestin2 compared with wild-type PTH1R. NHERF1 inhibited beta-arrestin2 binding to wtPTH1R but had no effect on beta-arrestin2 association with pdPTH1R. Such an effect may protect against PTH resistance or PTH1R down-regulation in cells harboring NHERF1. PMID- 19188334 TI - Dexmedetomidine vs midazolam for sedation of critically ill patients: a randomized trial. AB - CONTEXT: Gamma-aminobutyric acid receptor agonist medications are the most commonly used sedatives for intensive care unit (ICU) patients, yet preliminary evidence indicates that the alpha(2) agonist dexmedetomidine may have distinct advantages. OBJECTIVE: To compare the efficacy and safety of prolonged sedation with dexmedetomidine vs midazolam for mechanically ventilated patients. DESIGN, SETTING, AND PATIENTS: Prospective, double-blind, randomized trial conducted in 68 centers in 5 countries between March 2005 and August 2007 among 375 medical/surgical ICU patients with expected mechanical ventilation for more than 24 hours. Sedation level and delirium were assessed using the Richmond Agitation Sedation Scale (RASS) and the Confusion Assessment Method for the ICU. INTERVENTIONS: Dexmedetomidine (0.2-1.4 microg/kg per hour [n = 244]) or midazolam (0.02-0.1 mg/kg per hour [n = 122]) titrated to achieve light sedation (RASS scores between -2 and +1) from enrollment until extubation or 30 days. MAIN OUTCOME MEASURES: Percentage of time within target RASS range. Secondary end points included prevalence and duration of delirium, use of fentanyl and open label midazolam, and nursing assessments. Additional outcomes included duration of mechanical ventilation, ICU length of stay, and adverse events. RESULTS: There was no difference in percentage of time within the target RASS range (77.3% for dexmedetomidine group vs 75.1% for midazolam group; difference, 2.2% [95% confidence interval {CI}, -3.2% to 7.5%]; P = .18). The prevalence of delirium during treatment was 54% (n = 132/244) in dexmedetomidine-treated patients vs 76.6% (n = 93/122) in midazolam-treated patients (difference, 22.6% [95% CI, 14% to 33%]; P < .001). Median time to extubation was 1.9 days shorter in dexmedetomidine-treated patients (3.7 days [95% CI, 3.1 to 4.0] vs 5.6 days [95% CI, 4.6 to 5.9]; P = .01), and ICU length of stay was similar (5.9 days [95% CI, 5.7 to 7.0] vs 7.6 days [95% CI, 6.7 to 8.6]; P = .24). Dexmedetomidine-treated patients were more likely to develop bradycardia (42.2% [103/244] vs 18.9% [23/122]; P < .001), with a nonsignificant increase in the proportion requiring treatment (4.9% [12/244] vs 0.8% [1/122]; P = .07), but had a lower likelihood of tachycardia (25.4% [62/244] vs 44.3% [54/122]; P < .001) or hypertension requiring treatment (18.9% [46/244] vs 29.5% [36/122]; P = .02). CONCLUSIONS: There was no difference between dexmedetomidine and midazolam in time at targeted sedation level in mechanically ventilated ICU patients. At comparable sedation levels, dexmedetomidine-treated patients spent less time on the ventilator, experienced less delirium, and developed less tachycardia and hypertension. The most notable adverse effect of dexmedetomidine was bradycardia. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00216190 Published online February 2, 2009 (doi:10.1001/jama.2009.56). PMID- 19188336 TI - Induction of overexpression of the 27- and 70-kDa heat shock proteins by bicyclol attenuates concanavalin A-Induced liver injury through suppression of nuclear factor-kappaB in mice. AB - Heat shock proteins (HSPs) are molecular chaperones critical for cell survival under adverse environmental conditions and for normal cellular homeostasis. Bicyclol, a novel antihepatitis drug, has been shown to protect against liver injury in animals. However, it is unclear how bicyclol protects against liver injury. We recently found that bicyclol is an inducer of HSPs. We wondered whether bicyclol regulated the expression of HSPs to produce a liver protection in vivo. Thus, this study was designed to address these questions using a mouse model with concanavalin A (ConA)-induced liver injury. Oral administration of bicyclol markedly alleviated ConA-caused liver injury in mice as indicated by the reduction of serum aminotransferases, liver necrosis, and the release of cytochrome c and apoptosis-inducing factor from mitochondria and hepatic DNA fragmentation. Correlated with this, bicyclol induced the increase of mRNA and protein levels of hepatic 27- and 70-kDa HSPs (HSP27 and HSP70) in the mice. Correspondingly, the elevated HSP27 and HSP70 suppressed inhibitor kappaB degradation and nuclear factor kappaB (NF-kappaB) activation that were caused by ConA. The protective effects of bicyclol on ConA-induced mouse liver injury were markedly attenuated by quercetin, an inhibitor of HSPs synthesis. Our results suggest that the antihepatitis drug bicyclol may protect against liver injury by inducing the expression of hepatic HSP27 and HSP70 and consequently inhibit the transcription factor NF-kappaB-mediated apoptosis and necrosis in liver tissue. PMID- 19188337 TI - Dynamic patterns of histone methylation are associated with ontogenic expression of the Cyp3a genes during mouse liver maturation. AB - Human cytochrome P450 3A (CYP3A) members are major drug-metabolizing enzymes in the liver. Two genes, CYP3A4 and CYP3A7, exhibit a developmental switch in gene expression during liver maturation. CYP3A4 is mainly expressed in adults, whereas CYP3A7 is dominantly expressed during the fetal and neonatal stages. Their ontogenic expression results in developmentally related changes in the capacity to metabolize endogenous and exogenous compounds. Thus, it is desirable to understand the mechanisms controlling the developmental switch. Mice also exhibit a developmental switch between Cyp3a16 (neonatal isoform) and Cyp3a11 (adult isoform) and may serve as a model to study the mechanisms controlling the developmental switch. Because the epigenetic code (e.g., DNA methylation and histone modifications) is implicated in regulating gene expression and cellular differentiation during development, the current study determined the status of DNA methylation, histone-3-lysine-4 dimethylation (H3K4me2) and histone-3-lysine 27 trimethylation (H3K27me3) around the mouse Cyp3a locus at various developmental ages from prenatal, through neonatal, to young adult. DNA was not hypermethylated in the Cyp3a locus at any age. However, increases in Cyp3a16 expression in neonatal livers and Cyp3a11 in adult livers were associated with increases of H3K4me2. Suppression of Cyp3a16 expression in adult livers coincided with decreases of H3K4me2 and increases of H3K27me3 around Cyp3a16. In conclusion, histone modifications of H3K4me2 and H3K27me3 are dynamically changed in a locus-specific manner along the Cyp3a locus. Developmental switch between Cyp3a11 and Cyp3a16 gene expression seems to be due to dynamic changes of histone modifications during postnatal liver maturation. PMID- 19188339 TI - Reported cancer screening practices of nephrologists: results from a national survey. AB - BACKGROUND: Cancer is becoming increasingly recognized as a complication of chronic kidney disease (CKD), and screening is a widely used strategy to reduce cancer risk and improve outcomes. This study aimed to describe cancer screening practices by nephrologists in Australia and New Zealand, and to identify reasons for a positive recommendation to screen. METHODS: Questionnaires were posted electronically to all nephrologists who were in adult clinical practice in 2007 using the Australia-New Zealand Society of Nephrology register. The survey instrument assessed nephrologists' reported practice towards colorectal, breast and cervical cancer screening across all stages of CKD (CKD 1-5, dialysis and transplantation). RESULTS: Of the 250 eligible members, 131 (52%) participated, with over 75% recommending breast cancer screening (usually 1-2 yearly using combined mammography and breast-self examination), 48% recommending colorectal cancer screening (1-2 yearly faecal occult blood test) and 86% recommending cervical cancer screening (1-2 yearly conventional cytology). Recommendations to screen did not vary appreciably with CKD status. Recommended screening strategies were more frequent, included more invasive tests, and were targeted at a broader age range than national cancer screening programmes in the general population. Increased cancer prevalence and cancer-specific mortality benefits for screening were the most commonly reported and influential criteria for making a positive recommendation. CONCLUSIONS: Most nephrologists recommend breast and cervical, but not colorectal cancer screening in people with CKD. Despite the lack of trial based evidence of benefits of screening in this setting, recommended screening practices by nephrologists are more intense than for the general population. Increased disease prevalence appears to be the most influential factor for making a positive recommendation to screen in the CKD population even though this is not an internationally accepted criterion for a screening programme, and is not relevant for breast cancer, which is not increased in the CKD population. PMID- 19188338 TI - Lithium increases synapse formation between hippocampal neurons by depleting phosphoinositides. AB - The mood-stabilizing effects of lithium are well documented, although its mechanism of action remains unknown. Increases in gray matter volume detected in patients with bipolar disorder who were treated with lithium suggest that changes in the number of synapses might underlie its therapeutic effects. We investigated the effects of lithium on the number of synaptic connections between hippocampal neurons in culture. Confocal imaging of neurons expressing postsynaptic density protein 95 fused to green fluorescent protein (PSD95-GFP) enabled visualization of synaptic sites. PSD95-GFP fluorescent puncta represented functional synapses, and lithium (4 h, 5 mM) increased their number by 150 +/- 12%. The increase was time- and concentration-dependent (EC(50) = 1.0 +/- 0.6 mM). Lithium induced a parallel increase in the presynaptic marker synaptophysin-GFP. Valproic acid, another mood stabilizer, also increased the number of fluorescent puncta at a clinically relevant concentration. Inhibition of postsynaptic glutamate receptors or presynaptic inhibition of neurotransmitter release significantly reduced lithium-induced synapse formation, indicating that glutamatergic synaptic transmission was required. Pretreatment with exogenous myo-inositol inhibited synapse formation, demonstrating that depletion of inositol was necessary to increase synaptic connections. In contrast, inhibition of glycogen synthase kinase 3beta did not mimic lithium-induced synapse formation. Pharmacological and lipid reconstitution experiments showed that new synapses formed as a result of depletion of phosphatidylinositol-4-phosphate rather than a build-up of polyphosphoinositides or changes in the activity of phospholipase C, protein kinase C, or phosphatidylinositol-3-kinase. Increased synaptic connections may underlie the mood-stabilizing effects of lithium in patients with bipolar disorder and could contribute to the convulsions produced by excessive doses of this drug. PMID- 19188340 TI - Rapid adaptation of the intrarenal resistance index after living donor kidney transplantation. AB - BACKGROUND: Limited data exist concerning changes of renal perfusion directly after kidney transplantation. Colour-coded duplex sonography is the accepted method to assess kidney perfusion after transplantation. A widely used, although unspecific, Doppler parameter is the intrarenal resistance index (RI). The aim of this study was to clarify the influence of different patient- and procedure related factors on RI before and immediately after living kidney transplantation. METHODS: In a prospective study, 80 living kidney transplantation donor-recipient pairs were included. RI was measured in the donor 1 to 3 days before nephrectomy and in the recipient during the first hour after transplantation to examine the influence of age, heart rate, duration of cold and warm ischaemia time and immunosuppressive medications. RESULTS: Mean RI did not differ between donors and recipients. RI correlated with age, both in donors (r = 0.58, P < 0.001) and recipients (r = 0.39, P < 0.001). In recipients, 10 or more years younger than their donors (n = 24), an average decrease of 0.05 in RI compared to the donors' value was observed (P = 0.01). Heart rate, cold and warm ischaemia time and immunosuppressive medications had no influence on the recipient RI. In patients with delayed graft function, a significant increase in RI within 14 days was observed. However, the initial RI was not predictive of graft function. CONCLUSIONS: The transplanted kidney seems to be able to adjust its RI within a short time despite several potential harmful factors that can occur during the transplantation. PMID- 19188341 TI - Oral charcoal adsorbent (AST-120) prevents progression of cardiac damage in chronic kidney disease through suppression of oxidative stress. AB - BACKGROUND: Chronic kidney disease (CKD) is an important risk factor for cardiovascular disease (CVD). Increased oxidative stress plays a role in the pathogenesis of CVD in CKD patients. The oral charcoal adsorbent AST-120 attenuates the progression of CKD possibly by removing uraemic toxins such as indoxyl sulfate (IS), and reduces oxidative stress. We investigated the relationship between oxidative stress and cardiac damage in CKD and its prevention by AST-120. METHODS: Male Lewis rats were administered adriamycin at 8 weeks of age, and the right kidney was removed at 12 weeks of age. From 14 weeks of age, the rats were treated daily with AST-120 (n = 8) or were untreated (control group, n = 8). At 34 weeks of age, the rats were killed and urinary and blood biochemical tests as well as cardiac histological analyses were performed. RESULTS: At 14 weeks of age, there were no significant differences in blood pressure, renal function (creatinine clearance: 1.54 +/- 0.28 mL/min versus 1.60 +/- 0.22 mL/min), oxidative stress markers or other biochemical data between the control and AST-120 groups. At 34 weeks, despite similar blood pressure and renal function (creatinine clearance: 0.78 +/- 0.46 mL/min versus 0.75 +/- 0.54 mL/min), serum concentrations of IS and urinary excretion of 8 hydroxydeoxyguanosine (8-OHdG), acrolein and IS were significantly lower in the AST-120 group than in the control group. Heart volume, left ventricular volume and cardiac fibrosis were significantly smaller in the experimental AST-120 group than in the control group. Immunohistological analysis revealed that the numbers of 8-OHdG- and acrolein-positive cardiomyocytes and the degrees of myocardial and perivascular fibrosis were ameliorated by AST-120 administration. The myocardial fibrosis score was significantly associated with the 8-OHdG- (r = 0.848, P < 0.001) and acrolein-positive (r = 0.812, P < 0.001) cell scores. The perivascular fibrosis score was also significantly associated with the 8-OHdG- (r = 0.906, P < 0.0001) and acrolein-positive (r = 0.789, P < 0.001) cell scores. CONCLUSIONS: Oxidative stress is suggested to play a key role in the development of cardiac hypertrophy and fibrosis in CKD. AST-120 may suppress oxidative stress and reduce cardiac damage in CKD. PMID- 19188342 TI - Fluvastatin prevents podocyte injury in a murine model of HIV-associated nephropathy. AB - BACKGROUND: Recent studies have reported that statins have renoprotective effects, independent from lowering plasma cholesterol. In this study, we examined whether statins were beneficial in a murine model of HIV-associated nephropathy (HIVAN). METHODS: We used conditional transgenic mice that express one of the HIV 1 accessory genes, vpr, selectively in podocytes using podocin promoter and the Tet-on system. These mice develop aggressive collapsing focal segmental glomerular sclerosis with massive proteinuria and deterioration of renal function within 4 weeks following heminephrectomy and doxycycline administration. Fluvastatin was administrated simultaneously with doxycycline, and the effect was compared with untreated controls after 4 weeks. RESULTS: Fluvastatin at 10 mg/kg/day significantly decreased urinary albumin excretion (87 versus 11 mg/day, P < 0.01) and glomerular sclerosis (2.4 versus 1.0, P < 0.01, assessed by semi quantitative scoring: 0-4). Fluvastatin also decreased serum creatinine and total cholesterol, but these differences were not statistically significant (0.36 versus 0.32 mg/dl, P = 0.35; 492 versus 378 mg/dl, P = 0.11, respectively). Phenotypic changes in podocytes, as indicated by the downregulation of nephrin, Wilms' tumour 1 and synaptopodin, along with upregulation of proliferating cell nuclear antigen, were attenuated by fluvastatin, suggesting its protective effects against podocyte injuries. In cultured podocytes, angiotensin II treatment decreased nephrin expression to 13% of basal levels, which was reversed to 58% by adding fluvastatin. CONCLUSIONS: In conclusion, fluvastatin was effective in treating experimental HIVAN. The beneficial effect of this drug might be caused, in part, by preserving nephrin expression in podocytes against angiotensin II-mediated injury. PMID- 19188344 TI - Survey of the characteristics of cats owned by households in the UK and factors affecting their neutered status. AB - Data were collected on 1260 cats owned by a random sample of UK households in 2007, by means of telephone questionnaires, which were completed by 33 per cent of the households contacted. Cats were owned by 26 per cent of these households, 42 per cent of which owned more than one cat. The owners of 622 female cats reported a total of 110 unplanned litters of kittens. Excluding cats with an unknown neutered status 92 per cent of the cats aged six months or more were neutered, but only 66 per cent of the cats aged six to 12 months were neutered. Multivariable logistic regression, based on data collected for one randomly selected cat aged four months or more in each cat-owning household, was used to identify factors affecting the cats' neutered status. Analysis of 48 sexually entire cats and 501 neutered cats showed that cats aged 10 months or more, with indoor access, that had been vaccinated within the last year or were currently registered with a veterinary practice, were more likely to be neutered than cats aged less than 10 months, without indoor access, that had been vaccinated more than one year ago or were not registered with a veterinary practice. PMID- 19188345 TI - Bilateral laryngeal paralysis associated with hepatic dysfunction and hepatic encephalopathy in six ponies and four horses. AB - Six ponies and four horses with a mean (sd) age of 15.9 (6.0) years developed sudden-onset bilateral laryngeal paralysis (BLP) in association with hepatic dysfunction. Nine of them had been referred for the investigation of respiratory distress, and one pony had been referred for weight loss before BLP developed. Nine of the animals had clinicopathological evidence of liver disease, and nine had histological evidence of liver disease. All of the animals had one or more of the following: hepatic encephalopathy (in eight), hyperammonaemia (in six) and endoscopic evidence of BLP (in nine). Three of the animals had signs of pituitary pars intermedia dysfunction, a diagnosis supported in two by endocrine function testing, and in two by histopathological examination. Histopathological examination of the intrinsic laryngeal musculature and recurrent laryngeal nerves of four of the horses and of the region of the nucleus ambiguus of two did not reveal any abnormalities. Three of the animals were euthanased after they had first been examined, and one improved temporarily before the condition recurred. A temporary tracheostomy was performed in six of the animals, five of which subsequently died or were euthanased; one pony recovered. PMID- 19188346 TI - Effects of intramuscular acepromazine and diazepam on tear production in rabbits. AB - Sixteen New Zealand white rabbits were divided randomly into two groups of eight. Schirmer II tear tests were performed on all 16 and then eight received an intramuscular dose of 1 mg/kg acepromazine and the other eight received an intramuscular dose of 1 mg/kg diazepam. The Schirmer tests were repeated after 15 and 25 minutes. There was a significant reduction in tear production by the rabbits treated with acepromazine, but no significant change in tear production by the rabbits treated with diazepam. PMID- 19188347 TI - Emerging canine angiostrongylosis in northern England: five fatal cases. PMID- 19188348 TI - Abscessation, osteomyelitis and fracture of the sternum in a cow. PMID- 19188349 TI - Chlamydophila psittaci in wild birds in the UK. PMID- 19188350 TI - Availability of leptospirosis vaccine. PMID- 19188351 TI - Cost and responsibility sharing. PMID- 19188352 TI - Autochthonous case of Dirofilaria repens in a dog in the Netherlands. PMID- 19188354 TI - Generation of antibody responses to pneumococcal capsular polysaccharides is independent of CD1 expression in mice. AB - Streptococcus pneumoniae is a bacterial microorganism that frequently causes serious infection, particularly in children and the elderly. Protection against infection with S. pneumoniae is based mainly on the generation of antibodies to the pneumococcal capsular polysaccharides (caps-PS), but the mechanisms responsible for the generation of anticapsular antibodies remain incompletely understood. The aim of the present study was to evaluate the role of CD1 restricted T cells in the antibody response to caps-PS. When immunized with Pneumo23, wild-type mice and CD1 knockout mice on BALB/c and C57BL/6 backgrounds generated immunoglobulin M (IgM) and IgG antibody responses to soluble caps-PS that were comparable. Similar results were obtained after immunization with heat inactivated S. pneumoniae. The IgM and IgG antibody response of wild-type mice to Pneumo23 was not affected by an antagonizing monoclonal anti-CD1 antibody treatment. In summary, our data provide evidence that the antibody response to caps-PS is generated independently of CD1 expression. PMID- 19188353 TI - Induction of persistent colitis by a human commensal, enterotoxigenic Bacteroides fragilis, in wild-type C57BL/6 mice. AB - Enterotoxigenic Bacteroides fragilis (ETBF) causes diarrhea and is implicated in inflammatory bowel diseases and colorectal cancer. The only known ETBF virulence factor is the Bacteroides fragilis toxin (BFT), which induces E-cadherin cleavage, interleukin-8 secretion, and epithelial cell proliferation. A murine model for ETBF has not been characterized. Specific pathogen-free (SPF) C57BL/6J or germfree 129S6/SvEv mice were orally inoculated with wild-type ETBF (WT-ETBF) strains, a nontoxigenic WT strain of B. fragilis (WT-NTBF), WT-NTBF overexpressing bft (rETBF), or WT-NTBF overexpressing a biologically inactive mutated bft (rNTBF). In SPF and germfree mice, ETBF caused colitis but was lethal only in germfree mice. Colonic histopathology demonstrated mucosal thickening with inflammatory cell infiltration, crypt abscesses, and epithelial cell exfoliation, erosion, and ulceration. SPF mice colonized with rETBF mimicked WT ETBF, whereas rNTBF caused no histopathology. Intestinal epithelial E-cadherin was rapidly cleaved in vivo in WT-ETBF-colonized mice and in vitro in intestinal tissues cultured with purified BFT. ETBF mice colonized for 16 months exhibited persistent colitis. BFT did not directly induce lymphocyte proliferation, dendritic cell stimulation, or Toll-like receptor activation. In conclusion, WT ETBF induced acute then persistent colitis in SPF mice and rapidly lethal colitis in WT germfree mice. Our data support the hypothesis that chronic colonization with the human commensal ETBF can induce persistent, subclinical colitis in humans. PMID- 19188355 TI - Modelling of infection by enteropathogenic Escherichia coli strains in lineages 2 and 4 ex vivo and in vivo by using Citrobacter rodentium expressing TccP. AB - Enteropathogenic Escherichia coli (EPEC) strains colonize the human gut mucosa via attaching-and-effacing (A/E) lesion formation, while in vitro they employ diverse strategies to trigger actin polymerization. Strains belonging to the EPEC 1 lineage trigger strong actin polymerization via tyrosine phosphorylation of the type III secretion system (T3SS) effector Tir, recruitment of Nck, and activation of N-WASP. Strains belonging to EPEC-2 and EPEC-4 can trigger strong actin polymerization by dual mechanisms, since while employing the Tir-Nck pathway they can additionally activate N-WASP via the T3SS effectors TccP2 and TccP, respectively. It is currently not known if the ability to trigger actin polymerization by twin mechanisms increases in vivo virulence or fitness. Since mice are resistant to EPEC infection, in vivo studies are frequently done using the murine model pathogen Citrobacter rodentium, which shares with EPEC-1 strains the ability to induce A/E lesions and trigger strong actin polymerization via the Tir:Nck pathway. In order to model infections with EPEC-2 and EPEC-4, we constructed C. rodentium strains expressing TccP. Using a mouse intestinal in vitro organ culture model and oral gavage into C57BL/6 mice, we have shown that TccP can cooperate with Tir of C. rodentium. The recombinant strains induced typical A/E lesions ex vivo and in vivo. Expression of TccP did not alter C. rodentium colonization dynamics or pathology. In competition with the wild-type strain, expression of TccP in C. rodentium did not confer a competitive advantage. PMID- 19188356 TI - RICK promotes inflammation and lethality after gram-negative bacterial infection in mice stimulated with lipopolysaccharide. AB - RICK (receptor-interacting protein-like interacting caspase-like apoptosis regulatory protein kinase), a serine-threonine kinase, functions downstream of the pattern recognition receptors Nod1 and Nod2 to mediate NF-kappaB and mitogen activated protein kinase (MAPK) activation in response to specific microbial stimuli. However, the function of RICK in the recognition and host defense of gram-negative bacteria remains poorly understood. We report here that infection of wild-type and RICK-deficient macrophages with Pseudomonas aeruginosa and Escherichia coli elicited comparable activation of NF-kappaB and MAPKs as well as secretion of proinflammatory cytokines. However, production of interleukin 6 (IL 6) and IL-1beta induced by these gram-negative bacteria was impaired in RICK deficient macrophages when the cells had previously been stimulated with lipopolysaccharide (LPS) or E. coli. The diminished proinflammatory response of RICK-deficient macrophages to bacteria was associated with reduced activation of NF-kappaB and MAPKs. Importantly, mutant mice deficient in RICK were less susceptible than wild-type mice to P. aeruginosa infection when the animals had previously been stimulated with LPS. The reduced lethality of RICK-deficient mice infected with P. aeruginosa was independent of pathogen clearance but was associated with diminished production of proinflammatory molecules in vivo. These results demonstrate that RICK contributes to the induction of proinflammatory responses and susceptibility to gram-negative bacteria after exposure to LPS, a condition that is associated with reduced Toll-like receptor signaling. PMID- 19188357 TI - Interconnections between Sigma B, agr, and proteolytic activity in Staphylococcus aureus biofilm maturation. AB - Staphylococcus aureus is a proficient biofilm former on host tissues and medical implants. We mutagenized S. aureus strain SH1000 to identify loci essential for ica-independent mechanisms of biofilm maturation and identified multiple insertions in the rsbUVW-sigB operon. Following construction and characterization of a sigB deletion, we determined that the biofilm phenotype was due to a lack of sigma factor B (SigB) activity. The phenotype was conserved in a sigB mutant of USA300 strain LAC, a well-studied community-associated methicillin-resistant S. aureus isolate. We determined that agr RNAIII levels were elevated in the sigB mutants, and high levels of RNAIII expression are known to have antibiofilm effects. By introducing an agr mutation into the SH1000 or LAC sigB deletion strain, S. aureus regained biofilm capacity, indicating that the biofilm phenotype was agr dependent. Protease activity is linked to agr activity and ica independent biofilm formation, and we observed that the protease inhibitors phenylmethylsulfonyl fluoride and alpha-macroglobulin could reverse the sigB biofilm defect. Similarly, inactivating genes encoding both the aureolysin and Spl extracellular proteases in the sigB mutant restored biofilm capacity. Due to the growing link between murein hydrolase activity and biofilm maturation, autolysin zymography was performed, which revealed an altered profile in the sigB mutant; again, the phenotype could be repaired through protease inactivation. These findings indicate that the lack of SigB activity results in increased RNAIII expression, thus elevating extracellular protease levels and altering the murein hydrolase activity profile. Altogether, our observations demonstrate that SigB is an essential regulator of S. aureus biofilm maturation. PMID- 19188358 TI - Arginine-induced germ tube formation in Candida albicans is essential for escape from murine macrophage line RAW 264.7. AB - The opportunistic fungal pathogen Candida albicans is a part of the normal flora but it also causes systemic candidiasis if it reaches the bloodstream. Upon being phagocytized by macrophages, an important component of innate immunity, C. albicans rapidly upregulates a set of arginine biosynthetic genes. Arginine, urea, and CO2 induced hyphae in a density-dependent manner in wild-type, cph1/cph1, and rim101/rim101 strains but not in efg1/efg1 or cph1/cph1 efg1/efg1 strains. Arginase (Car1p) converts arginine to urea, which in turn is degraded by urea amidolyase (Dur1,2p) to produce CO2, a signal for hyphal switching. We used a dur1,2/dur1,2 mutant (KWN6) and the complemented strain, KWN8 (dur1,2/dur1,2::DUR1,2/DUR1,2) to study germ tube formation. KWN6 could not make germ tubes in the presence of arginine or urea but did in the presence of 5% CO(2), which bypasses Dur1,2p. We also tested the effect of arginine on the interaction between the macrophage line RAW 264.7 and several strains of C. albicans. Arginine activated an Efg1p-dependent yeast-to-hypha switch, enabling wild-type C. albicans and KWN8 to escape from macrophages within 6 h, whereas KWN6 was defective in this regard. Additionally, two mutants that cannot synthesize arginine, BWP17 and SN152, were defective in making hyphae inside the macrophages, whereas the corresponding arginine prototrophs, DAY286 and SN87, formed germ tubes and escaped from macrophages. Therefore, metabolism of arginine by C. albicans controls hyphal switching and provides an important mechanism for escaping host defense. PMID- 19188359 TI - Different pathologies but equal levels of responsiveness to the recombinant F1 and V antigen vaccine and ciprofloxacin in a murine model of plague caused by small- and large-particle aerosols. AB - Presently there is a significant effort to develop and evaluate vaccines and antibiotics against the potential bioterrorism agent Yersinia pestis. The animal models used to test these countermeasures involve the deposition of small particles within the lung. However, deliberate aerosol release of Y. pestis will generate both small and large inhalable particles. We report in this study that the pathogenesis patterns of plague infections caused by the deposition of 1- and 12-microm-particle aerosols of Y. pestis in the lower and upper respiratory tracts (URTs) of mice are different. The median lethal dose for 12-mum particles was 4.9-fold greater than that for 1-microm particles. The 12-microm-particle infection resulted in the degradation of the nasal mucosa and nasal-associated lymphoid tissue (NALT) plus cervical lymphadenopathy prior to bacteremic dissemination. Lung involvement was limited to secondary pneumonia. In contrast, the 1-microm-particle infection resulted in primary pneumonia; in 40% of mice, the involvement of NALT and cervical lymphadenopathy were observed, indicating entry via both URT lymphoid tissues and lungs. Despite bacterial deposition in the gastrointestinal tract, the involvement of Peyer's patches was not observed in either infection. Although there were major differences in pathogenesis, the recombinant F1 and V antigen vaccine and ciprofloxacin protected against plague infections caused by small- and large-particle aerosols. PMID- 19188360 TI - Independence of Anaplasma marginale strains with high and low transmission efficiencies in the tick vector following simultaneous acquisition by feeding on a superinfected mammalian reservoir host. AB - Strain superinfection occurs when a second pathogen strain infects a host already carrying a primary strain. Anaplasma marginale superinfection occurs when the second strain carries a variant repertoire different from that of the primary strain, and the epidemiologic consequences depend on the relative efficiencies of tick-borne transmission of the two strains. Following strain superinfection in the reservoir host, we tested whether the presence of two A. marginale (sensu lato) strains that differed in transmission efficiency altered the transmission phenotypes in comparison to those for single-strain infections. Dermacentor andersoni ticks were fed on animals superinfected with the Anaplasma marginale subsp. centrale vaccine strain (low transmission efficiency) and the A. marginale St. Maries strain (high transmission efficiency). Within ticks that acquired both strains, the St. Maries strain had a competitive advantage and replicated to significantly higher levels than the vaccine strain. The St. Maries strain was subsequently transmitted to naive hosts by ticks previously fed either on superinfected animals or on animals singly infected with the St. Maries strain, consistent with the predicted transmission phenotype of this strain and the lack of interference due to the presence of a competing low-efficiency strain. The vaccine strain was not transmitted by either singly infected or coinfected ticks, consistent with the predicted transmission phenotype and the lack of enhancement due to the presence of a high-efficiency strain. These results support the idea that the strain predominance in regions of endemicity is mediated by the intrinsic transmission efficiency of specific strains regardless of occurrence of superinfection. PMID- 19188361 TI - Modulation of cell wall structure and antimicrobial susceptibility by a Staphylococcus aureus eukaryote-like serine/threonine kinase and phosphatase. AB - It is well established that prokaryotes and eukaryotes alike utilize phosphotransfer to regulate cellular functions. One method by which this occurs is via eukaryote-like serine/threonine kinase (ESTK)- and phosphatase (ESTP) regulated pathways. The role of these enzymes in Staphylococcus aureus has not yet been examined. This resilient organism is a common cause of hospital-acquired and community-associated infections, infecting immunocompromised and immunocompetent hosts alike. In this study, we have characterized a major functional ESTK (STK) and ESTP (STP) in S. aureus and found them to be critical modulators of cell wall structure and susceptibility to cell wall-acting beta lactam antibiotics. By utilizing gene knockout strategies, we created S. aureus N315 mutants lacking STP and/or STK. The strain lacking both STP and STK displayed notable cell division defects, including multiple and incomplete septa, bulging, and irregular cell size, as observed by transmission electron microscopy. Mutants lacking STP alone displayed thickened cell walls and increased resistance to the peptidoglycan-targeting glycylglycine endopeptidase lysostaphin, compared to the wild type. Additionally, mutant strains lacking STK or both STK and STP displayed increased sensitivity to cell wall-acting cephalosporin and carbapenem antibiotics. Together, these results indicate that S. aureus STK- and STP-mediated reversible phosphorylation reactions play a critical role in proper cell wall architecture, and thus the modulation of antimicrobial resistance, in S. aureus. PMID- 19188362 TI - USP17 regulates Ras activation and cell proliferation by blocking RCE1 activity. AB - The proto-oncogene Ras undergoes a series of post-translational modifications at its carboxyl-terminal CAAX motif that are essential for its proper membrane localization and function. One step in this process is the cleavage of the CAAX motif by the enzyme Ras-converting enzyme 1 (RCE1). Here we show that the deubiquitinating enzyme USP17 negatively regulates the activity of RCE1. We demonstrate that USP17 expression blocks Ras membrane localization and activation, thereby inhibiting phosphorylation of the downstream kinases MEK and ERK. Furthermore, we show that this effect is caused by the loss of RCE1 catalytic activity as a result of its deubiquitination by USP17. We also show that USP17 and RCE1 co-localize at the endoplasmic reticulum and that USP17 cannot block proliferation or Ras membrane localization in RCE1 null cells. These studies demonstrate that USP17 modulates Ras processing and activation, at least in part, by regulating RCE1 activity. PMID- 19188363 TI - Extracellular microfibrils in vertebrate development and disease processes. PMID- 19188364 TI - Identification of myo-inositol-3-phosphate synthase isoforms: characterization, expression, and putative role of a 16-kDa gamma(c) isoform. AB - Myo-inositol is an important constituent of membrane phospholipids and is a precursor for the phosphoinositide signaling pathway. It is synthesized from glucose 6-phosphate by myo-inositol-3-phosphate synthase (IP synthase), a homotrimer composed of a 68-kDa polypeptide in most mammalian tissues. It is a putative target for mood-stabilizing drugs such as lithium and valproate. Here, we show that the rat gene (Isyna1) encoding this enzyme generates a number of alternatively spliced transcripts in addition to the fully spliced form that encodes the 68-kDa subunit (the alpha isoform). Specifically, we identify a small 16-kDa subunit (the gamma(c) isoform) derived by an intron retention mechanism and provide evidence for its existence in rat tissues. The gamma(c) isoform is highly conserved in mammals, but it lacks the catalytic domain while retaining the NAD(+) binding domain. Both alpha and gamma(c) isoforms are predominantly expressed in many rat tissues and display apparent stoichiometry in purified enzyme preparations. An IP synthase polyclonal antibody not only detects the alpha and gamma(c) isoforms but also several other isoforms in pancreas, intestine, and testis suggesting that the holoenzyme is composed of unique subunits in various tissues. Interestingly, the alpha isoform is not expressed in the intestine. IP synthase activity assays using purified alpha and gamma(c) isoforms indicate that the latter negatively modulates alpha isoform activity, possibly by competing for NAD(+) molecules. Our findings have important ramifications for understanding the mood stabilization process and suggest that inositol biosynthesis is a highly regulated and dynamic process. PMID- 19188365 TI - The Endoplasmic Reticulum-associated Degradation of Transthyretin Variants Is Negatively Regulated by BiP in Mammalian Cells. AB - Amyloid fibril formation of mutant transthyretin (TTR) that causes familial amyloid polyneuropathy occurs in the extracellular space. Thus, secretion of TTR variants contributes to the pathogenesis of amyloidosis. However, the molecular mechanisms underlying the endoplasmic reticulum (ER) exit or retention and subsequent degradation of TTR variants remain unclear. Here, we demonstrated that the nonsecreted TTR variants, such as D18G TTR and amyloidogenic TTRs with introduced monomeric mutation (M-TTRs), stably interact with the ER chaperone BiP in mammalian cells. These proteins were co-secreted with the secreted form of BiP in which the KDEL signal was removed, indicating that BiP partially contributes to the ER retention of nonsecreted TTR variants. More interestingly, the degradation efficiency of nonsecreted TTRs was increased when BiP was down regulated by small interfering RNA. Thus, BiP protects the TTR variants from immediate degradation. Additionally, we showed that the stability of nonsecreted TTR variants is not disturbed in the coat complex II-deficient conditions, which are enough to inhibit the ER export of secreted TTR variants, including wild-type TTR. Therefore, the post-ER retrieval mechanism might not contribute to the ER associated degradation of nonsecreted TTR variants. These findings suggest that the affinity to the ER-resident protein BiP regulates the fate of TTR variants in the ER. PMID- 19188366 TI - Two dermatan sulfate epimerases form iduronic acid domains in dermatan sulfate. AB - A second dermatan sulfate epimerase (DS-epi2) was identified as a homolog of the first epimerase (DS-epi1), which was previously described by our group. DS-epi2 is 1,222 amino acids long and has an approximately 700-amino acid N-terminal epimerase domain that is highly conserved between the two enzymes. In addition, the C-terminal portion is predicted to be an O-sulfotransferase domain. In this study we found that DS-epi2 has epimerase activity, which involves conversion of d-glucuronic acid to l-iduronic acid (EC 5.1.3.19), but no O-sulfotransferase activity was detected. In dermatan sulfate, iduronic acid residues are either clustered together in blocks or alternating with glucuronic acid, forming hybrid structures. By using a short interfering RNA approach, we found that DS-epi2 and DS-epi1 are both involved in the biosynthesis of the iduronic acid blocks in fibroblasts and that DS-epi2 can also synthesize the hybrid structures. Both iduronic acid-containing domains have been shown to bind to several growth factors, many of which have biological roles in brain development. DS-epi2 has been genetically linked to bipolar disorder, which suggests that the dermatan sulfate domains generated by a defective enzyme may be involved in the etiology of the disease. PMID- 19188367 TI - A function for the RING finger domain in the allosteric control of MDM2 conformation and activity. AB - The MDM2 oncoprotein plays multiple regulatory roles in the control of p53 dependent gene expression. A picture of MDM2 is emerging where structurally discrete but interdependent functional domains are linked through changes in conformation. The domain structure includes: (i) a hydrophobic pocket at the N terminus of MDM2 that is involved in both its transrepressor and E3-ubiqutin ligase functions, (ii) a central acid domain that recognizes a ubiquitination signal in the core DNA binding domain of p53, and (iii) a C-terminal C2H2C4 RING finger domain that is required for E2 enzyme-binding and ATP-dependent molecular chaperone activity. Here we show that the binding affinity of MDM2s hydrophobic pocket can be regulated through the RING finger domain and that increases in pocket affinity are reflected by a gain in MDM2 transrepressor activity. Thus, mutations within the RING domain that affect zinc coordination, but not one that inhibits ATP binding, produce MDM2 proteins that have a higher affinity for the BOX-I transactivation domain of p53 and a reduced I(0.5) for p53 transrepression. An allosteric model for regulation of the hydrophobic pocket is supported by differences in protein conformation and pocket accessibility between wild-type and the RING domain mutant MDM2 proteins. Additionally the data demonstrate that the complex relationship between different domains of MDM2 can impact on the efficacy of anticancer drugs directed toward its hydrophobic pocket. PMID- 19188368 TI - Evolutionary basis for the coupled-domain motions in Thermus thermophilus leucyl tRNA synthetase. AB - Aminoacyl-tRNA synthetases are multidomain proteins that catalyze the covalent attachment of amino acids to their cognate transfer RNA. Various domains of an aminoacyl-tRNA synthetase perform their specific functions in a highly coordinated manner to maintain high accuracy in protein synthesis in cells. The coordination of their function, therefore, requires communication between domains. In this study we explored the relevance of enzyme motion in domain domain communications. Specifically, we attempted to probe whether the communication between distantly located domains of a multidomain protein is accomplished through a coordinated movement of structural elements. We investigated the collective motion in Thermus thermophilus leucyl-tRNA synthetase by studying the low frequency normal modes. We identified the mode that best described the experimentally observed conformational changes of T. thermophilus leucyl-tRNA synthetase upon substrate binding and analyzed the correlated and anticorrelated motions between different domains. Furthermore, we used statistical coupling analysis to explore if the amino acid pairs and/or clusters whose motions are thermally coupled have also coevolved. Our study demonstrates that a small number of residues belong to the category whose coupled thermal motions correspond to evolutionary coupling as well. These residue clusters constitute a distinguished set of interacting networks that are sparsely distributed in the domain interface. Residues of these networking clusters are within van der Waals contact, and we suggest that they are critical in the propagation of long range mechanochemical motions in T. thermophilus leucyl-tRNA synthetase. PMID- 19188369 TI - Different roles of C-terminal cassettes in the trafficking of full-length NR1 subunits to the cell surface. AB - N-Methyl-d-aspartate (NMDA) receptors are glutamate-gated ion channels composed of NR1 and NR2 subunits. When expressed alone, the most prevalent NR1 splice variant and all NR2 subunits are retained in the endoplasmic reticulum (ER), whereas other NR1 splice variants reach the cell surface to varying degrees. Because similar trafficking patterns have been seen for single transmembrane domain chimeric proteins with appended C termini of NMDA receptor subunits, these chimeric proteins have been used as a model for studying the mechanisms underlying the ER retention and surface trafficking of NMDA receptors. Using this approach, an RRR motif in the C1 cassette has been identified as a major ER retention signal present in NR1 subunits, and the surface localization of other NR1 splice variants has been explained by the absence of the C1 cassette or by the presence of a PDZ/coatomer protein complex II-binding domain in the C2' cassette. However, when we tested these conclusions using full-length NR1 constructs, a more complex role of the C-terminal cassettes in the trafficking of NR1 subunits emerged. Our experiments showed that two independent ER retention motifs in the C1 cassette, KKK and RRR, are the signals mediating ER retention of the full-length NR1 subunits and that the C2 cassette has an additional inhibitory effect on the forward trafficking of NR1 subunits. On the other hand, C0 and C2' cassettes had an enhancing effect on the trafficking of NR1 subunits to the cell surface. Our observations identify the unique roles of C-terminal cassettes in the trafficking of full-length NR1 subunits. PMID- 19188371 TI - Improving diabetes care and health measures among hispanics using community health workers: results from a randomized controlled trial. AB - The increasing prevalence of diabetes and obesity, growing health disparities, and shortage of bilingual and culturally trained health care professionals underscore the role of trained community health workers (CHWs) to provide economically sustainable and culturally relevant services. This prospective randomized design evaluated the relative effectiveness of a CHW intervention among Hispanic persons with newly diagnosed type 2 diabetes, as compared with usual clinic practice in three inner-city health centers. In sum, 189 Hispanic patients newly diagnosed with type 2 diabetes were randomly assigned to one of three 6-month diabetes management approaches--CHW, case management, and standard provider care--and assessed for diabetes-related health measures and clinical indicators at baseline and postintervention. Participants in the CHW group achieved greater improvements than did the controls in program measures: health status, emergency department utilization, dietary habits, physical activity, and medication adherence. They also had 2.9 times greater odds of decreasing body mass index. PMID- 19188370 TI - A toc159 import receptor mutant, defective in hydrolysis of GTP, supports preprotein import into chloroplasts. AB - The heterotrimeric Toc core complex of the chloroplast protein import apparatus contains two GTPases, Toc159 and Toc34, together with the protein-conducting channel Toc75. Toc159 and Toc34 are exposed at the chloroplast surface and function in preprotein recognition. Together, they have been shown to facilitate the import of photosynthetic proteins into chloroplasts in Arabidopsis. Consequently, the ppi2 mutant lacking atToc159 has a non-photosynthetic albino phenotype. Previous mutations in the conserved G1 and G3 GTPase motifs abolished the function of Toc159 in vivo by disrupting targeting of the receptor to chloroplasts. Here, we demonstrate that a mutant in a conserved G1 lysine (atToc159 K868R) defective in GTP binding and hydrolysis can target and assemble into Toc complexes. We show that atToc159 K868R can support protein import into isolated chloroplasts, albeit at lower preprotein binding and import efficiencies compared with the wild-type receptor. Considering the absence of measurable GTPase activity in the K868R mutant, we conclude that GTP hydrolysis at atToc159 is not strictly required for preprotein translocation. The data also indicate that preprotein import requires at least one additional GTPase other than Toc159. PMID- 19188372 TI - Estimates of intraclass correlation for variables related to behavioral HIV/STD prevention in a predominantly African American and Hispanic sample of young women. AB - Studies designed to evaluate HIV and STD prevention interventions often involve random assignment of groups such as neighborhoods or communities to study conditions (e.g., to intervention or control). Investigators who design group randomized trials (GRTs) must take the expected intraclass correlation coefficient (ICC) into account in sample size estimation to have adequate power; however, few published ICC estimates exist for outcome variables related to HIV and STD prevention. The Prevention Options for Women Equal Rights (POWER) study was a GRT designed to evaluate a campaign to increase awareness and use of condoms among young African American and Hispanic women. The authors used precampaign and postcampaign data from the POWER study to estimate ICCs (unadjusted and adjusted for covariates) for a variety of sexual behavior and other variables. To illustrate the impact of ICCs on power, the authors present sample-size calculations and demonstrate how ICCs of differing magnitude will affect estimates of required sample size. PMID- 19188375 TI - In vitro activities of fluconazole and voriconazole against clinical isolates of Candida spp. determined by disk diffusion testing in Turin, Italy. AB - The in vitro activities of fluconazole and voriconazole against 1,024 clinical isolates of Candida spp. were determined by the agar disk diffusion test using the Clinical and Laboratory Standards Institute (CLSI) M44-A guidelines. The results of this investigation demonstrated the broad-spectrum in vitro activity of voriconazole, relative to that of fluconazole, against yeasts tested, in particular fluconazole-resistant isolates, such as Candida krusei that showed high susceptibility to voriconazole. The situation in Turin, Italy, is quite similar to that of the rest of Italy, reflecting the worldwide trend. PMID- 19188376 TI - Azithromycin in Pseudomonas aeruginosa biofilms: bactericidal activity and selection of nfxB mutants. AB - Azithromycin (AZM) has shown promising results in the treatment of Pseudomonas aeruginosa chronic lung infections such as those occurring in cystic fibrosis (CF) patients. We evaluated the effect of hypermutation and alginate hyperproduction on the bactericidal activity and resistance development to AZM in P. aeruginosa biofilms. Strains PAO1, its microcA mutant (PAOMA), and their respective mutS-deficient hypermutable derivatives (PAOMS and PAOMSA) were used. Biofilms were incubated with several AZM concentrations for 1, 2, 4, or 7 days, and the numbers of viable cells were determined. During the first 2 days, AZM showed bactericidal activity for all the strains, but in extended AZM incubation for strain PAOMS and especially strain PAOMSA, a marked increased in the number of viable cells was observed, particularly at 4 microg/ml. Biofilms formed by the lineages recovered from the 7-day experiments showed enhanced AZM resistance. Furthermore, most of the independent lineages studied, including those obtained from biofilms treated with AZM concentrations as low as 0.5 microg/ml, showed MexCD-OprJ hyperexpression and mutations in nfxB. The role of nfxB mutation in AZM resistance was further confirmed through the characterization of nfxB and mexD knockout mutants. Results from this work show that, although AZM exhibits bactericidal activity against P. aeruginosa biofilms, resistant mutants are readily selected and that, furthermore, they frequently show cross-resistance to other unrelated antipseudomonal agents such as ciprofloxacin or cefepime but hypersusceptibility to others such as imipenem or tobramycin. Therefore, these results should help guide the selection of appropriate antipseudomonal therapies in CF patients under AZM maintenance treatment. PMID- 19188377 TI - blaCTX-M genes in escherichia coli strains from Croatian Hospitals are located in new (blaCTX-M-3a) and widely spread (blaCTX-M-3a and blaCTX-M-15) genetic structures. AB - CTX-M-producing Escherichia coli isolates from three Croatian hospitals were analyzed. All bla(CTX-M-15) genes and one bla(CTX-M-3a) gene resided in widely spread ISEcp1 transposition modules, but other bla(CTX-M-3a) genes were in a new configuration with two IS26 copies, indicating a new event of gene mobilization from a Kluyvera ascorbata genome. The study confirmed the role of the E. coli ST131 clonal group with IncFII-type plasmids in the spread of bla(CTX-M-15) and of IncL/M pCTX-M3-type plasmids in the dissemination of bla(CTX-M-3a). PMID- 19188378 TI - Breakpoints for susceptibility testing should not divide wild-type distributions of important target species. AB - The fluconazole MIC distributions for Candida glabrata from testing 34 different clinical isolates and performing 51 tests on a single isolate mirrored each other. Since what is perceived as biological variation in isolates without resistance mechanisms is mainly methodological variation, breakpoints which divide this distribution not only lack a sound biological basis but also result in poor reproducibility of susceptibility characterization. This makes 2, 4, 8, and possibly 16 microg/ml unsuitable breakpoints for C. glabrata and fluconazole. PMID- 19188379 TI - Low plasma membrane expression of the miltefosine transport complex renders Leishmania braziliensis refractory to the drug. AB - Miltefosine (hexadecylphosphocholine, MLF) is the first oral drug with recognized efficacy against both visceral and cutaneous leishmaniasis. However, some clinical studies have suggested that MLF shows significantly less efficiency against the cutaneous leishmaniasis caused by Leishmania braziliensis. In this work, we have determined the cellular and molecular basis for the natural MLF resistance observed in L. braziliensis. Four independent L. braziliensis clinical isolates showed a marked decrease in MLF sensitivity that was due to their inability to internalize the drug. MLF internalization in the highly sensitive L. donovani species requires at least two proteins in the plasma membrane, LdMT, a P type ATPase involved in phospholipid translocation, and its beta subunit, LdRos3. Strikingly, L. braziliensis parasites showed highly reduced levels of this MLF translocation machinery at the plasma membrane, mainly because of the low expression levels of the beta subunit, LbRos3. Overexpression of LbRos3 induces increased MLF sensitivity not only in L. braziliensis promastigotes but also in intracellular amastigotes. These results further highlight the importance of the MLF translocation machinery in determining MLF potency and point toward the development of protocols to routinely monitor MLF susceptibility in geographic areas where L. braziliensis might be prevalent. PMID- 19188380 TI - Transmission of IncN plasmids carrying blaCTX-M-1 between commensal Escherichia coli in pigs and farm workers. AB - CTX-M-1-producing Escherichia coli were isolated from 56 pigs, three farm personnel, two manure samples, and two air samples from two Danish pig farms where an association between prophylactic ceftiofur use and the occurrence of cephalosporin resistance was previously demonstrated. Human, animal, and environmental strains displayed high genetic diversity but harbored indistinguishable or closely related IncN plasmids carrying bla(CTX-M-1), indicating that IncN plasmids mediating cephalosporin resistance were transmitted between pigs and farm workers across multiple E. coli lineages. PMID- 19188381 TI - Gallium maltolate treatment eradicates Pseudomonas aeruginosa infection in thermally injured mice. AB - Gallium (Ga) is a semimetallic element that has demonstrated therapeutic and diagnostic-imaging potential in a number of disease settings, including cancer and infectious diseases. Gallium's biological actions stem from its ionic radius being almost the same as that of ferric iron (Fe(3+)), whereby it can replace iron (Fe) in Fe(3+)-dependent biological systems, such as bacterial and mammalian Fe transporters and Fe(3+)-containing enzymes. Unlike Fe(3+), ionic gallium (Ga(3+)) cannot be reduced, and when incorporated, it inactivates Fe(3+) dependent reduction and oxidation processes that are necessary for bacterial and mammalian cell proliferation. Most pathogenic bacteria require Fe for growth and function, and the availability of Fe in the host or environment can greatly enhance virulence. We examined whether gallium maltolate (GaM), a novel formulation of Ga, had antibacterial activity in a thermally injured acute infection mouse model. Dose-response studies indicated that a GaM dose as low as 25 mg/kg of body weight delivered subcutaneously was sufficient to provide 100% survival in a lethal P. aeruginosa-infected thermally injured mouse model. Mice treated with 100 mg/kg GaM had undetectable levels of Pseudomonas aeruginosa in their wounds, livers, and spleens, while the wounds of untreated mice were colonized with over 10(8) P. aeruginosa CFU/g of tissue and their livers and spleens were colonized with over 10(5) P. aeruginosa CFU/g of tissue. GaM also significantly reduced the colonization of Staphylococcus aureus and Acinetobacter baumannii in the wounds of thermally injured mice. Furthermore, GaM was also therapeutically effective in preventing preestablished P. aeruginosa infections at the site of the injury from spreading systemically. Taken together, our data suggest that GaM is potentially a novel antibacterial agent for the prevention and treatment of wound infections following thermal injury. PMID- 19188382 TI - Interactions between triazoles and amphotericin B in treatment of disseminated murine infection by Fusarium oxysporum. AB - We have evaluated and compared the efficacies of high doses of amphotericin B (AMB; 3 mg/kg of body weight/day), voriconazole (60 mg/kg), and posaconazole (PSC; 100 mg/kg) alone and combined in a murine model of disseminated infection by Fusarium oxysporum. The combination of AMB with PSC showed the best results, prolonging the survival of mice and reducing their organ fungal loads. This combination might constitute a therapeutic option for those infections where monotherapies fail. PMID- 19188383 TI - Chlamydia trachomatis laboratory strains versus recent clinical isolates: implications for routine microbicide testing. AB - A topical microbicide that women can use to prevent sexually transmitted diseases (STDs) is essential, and many microbicide candidates are being tested for activity against human immunodeficiency virus and other STDs, including Chlamydia trachomatis. Screening assays for assessing the activity of microbicides against C. trachomatis are typically done with laboratory-adapted strains, but it is possible that recent clinical isolates may have different susceptibilities to microbicides, as has been seen with Neisseria gonorrhoeae and Lactobacillus spp. (B. J. Moncla and S. L. Hillier, Sex. Transm. Dis. 32:491-494, 2005). We utilized three types of microbicides to help define this aspect of our assay to test microbicides against C. trachomatis in vitro. To simulate conditions of transmission, we used an assay that we previously developed in which we exposed chlamydial elementary bodies to microbicides prior to contact with epithelial cells. We first determined the toxicity of microbicides to the cells used to culture Chlamydia trachomatis in the assay and, if necessary, modified the assay to eliminate toxicity at the concentrations tested. We compared the sensitivities of recent clinical isolates of Chlamydia trachomatis versus laboratory strains of the same serovar and found major differences in sensitivity to nonoxynol-9 (non 9), but only minor differences were seen with the other microbicides. We thus conclude that when assessing activity of potential topical microbicides versus the obligate intracellular bacteria C. trachomatis, the use of recent clinical isolates may not be necessary to draw a conclusion about a microbicide's effectiveness. However, it is important to keep in mind that differences (like those seen with non-9) are possible and that clinical isolates could be included in later stages of testing. PMID- 19188384 TI - Contribution of the autolysin AtlA to the bactericidal activity of amoxicillin against Enterococcus faecalis JH2-2. AB - The bactericidal activity of amoxicillin was investigated against Enterococcus faecalis JH2-2 and against an isogenic mutant deficient in the production of the N-acetylglucosaminidase AtlA. Comparison of the two strains indicated that this autolysin contributes to killing by amoxicillin both in vitro and in a rabbit model of experimental endocarditis. PMID- 19188385 TI - Antistaphylococcal activity of dihydrophthalazine antifolates, a family of novel antibacterial drugs. AB - For a panel of 153 Staphylococcus aureus clinical isolates (including 13 vancomycin-intermediate or heterogeneous vancomycin-intermediate and 4 vancomycin resistant strains), MIC(50)s and MIC(90)s of three novel dihydrophthalazine antifolates, BAL0030543, BAL0030544, and BAL0030545, were 0.03 and 0.25 microg/ml, respectively, for methicillin-susceptible strains and 0.03 and 128 microg/ml), although rates of endogenous resistance development were much lower for the dihydrophthalazines than for trimethoprim. Single-step platings of naive staphylococci onto media containing dihydrophthalazine antifolates indicated considerable variability among strains with respect to preexistent subpopulations nonsusceptible to dihydrophthalazine antifolates. PMID- 19188386 TI - Efficacy of piperacillin combined with the Penem beta-lactamase inhibitor BLI-489 in murine models of systemic infection. AB - The in vivo efficacy of piperacillin in combination with the penem inhibitor BLI 489 was determined using acute lethal systemic infections in mice. On the basis of preliminary results with various ratios, a dosing ratio of 8:1 was found to be optimal for retention of enhanced efficacy. Piperacillin-BLI-489 dosed at an 8:1 ratio was efficacious against murine infections caused by class A (including extended-spectrum beta-lactamases), class C (AmpC), and class D beta-lactamase expressing pathogens. PMID- 19188387 TI - Steady-state pharmacokinetics of abacavir in plasma and intracellular carbovir triphosphate following administration of abacavir at 600 milligrams once daily and 300 milligrams twice daily in human immunodeficiency virus-infected subjects. AB - Abacavir (ABC) is administered either at 600 mg once daily (ABC 600 mg QD) or 300 mg twice daily (ABC 300 mg BID) in anti-human immunodeficiency virus (anti-HIV) combination therapy. Although ABC plasma pharmacokinetics following each regimen has been well defined, no study has directly compared the regimens with respect to pharmacokinetics of ABC's active intracellular anabolite, carbovir triphosphate (CBV-TP). In an open-label, two-period, crossover study, 34 HIV infected male and female subjects stabilized on antiretroviral regimens containing either ABC 600 mg QD or ABC 300 mg BID received their usual doses on days -1 and 1 and then switched regimens for days 2 to 11. Serial blood samples collected on days 1 and 11 were assayed for plasma ABC and intracellular CBV-TP concentrations using validated high-performance liquid chromatography-tandem mass spectrometry methods. Pharmacokinetic parameters were calculated using noncompartmental methods. Analysis of variance with a mixed-effect model was performed for treatment and gender comparisons. In 27 evaluable subjects, the regimens provided bioequivalent ABC daily areas under the concentration-time curve from 0 to 24 h (AUC(0-24)) and comparable CBV-TP concentrations at the end of the dosing interval (C(tau)). As expected, ABC QD resulted in 109% higher ABC maximum concentrations of drug in plasma (C(max)) than did ABC BID. ABC QD also resulted in 32% higher CBV-TP AUC(0-24) and 99% higher CBV-TP C(max) than did ABC BID. Females had a 38% higher weight-adjusted ABC AUC(0-24) and 81% higher weight adjusted CBV-TP AUC(0-24) than did males. Virologic suppression was maintained during regimen switch, and no tolerability differences between regimens were observed. In conclusion, this study showed that ABC 600 mg QD and ABC 300 mg BID regimens led to similar intracellular CBV-TP C(tau) values, thus providing pharmacokinetic support for the interchangeability of these two regimens. Women had higher intracellular CBV-TP exposure than did men. PMID- 19188389 TI - Sitafloxacin and garenoxacin may overcome the antibiotic resistance of Helicobacter pylori with gyrA mutation. PMID- 19188388 TI - Role of known molecular markers of resistance in the antimalarial potency of piperaquine and dihydroartemisinin in vitro. AB - Using a range of laboratory-adapted and genetically modified Plasmodium falciparum parasite isolates, we investigated the interaction between dihydroartemisinin and piperaquine (PIP), the individual components of an artemisinin combination therapy currently under development, in addition to the role of known drug resistance genes in parasite susceptibility in vitro. All but one parasite line investigated displayed an interaction of dihydroartemisinin and PIP that was antagonistic, although the degree of antagonism was isolate dependent. In terms of resistance markers, the pfcrt haplotypes CVIET and SVMNT were positively associated with reduced sensitivity to PIP, with parasites carrying the South American CQR (SVMNT) allele being generally less sensitive than CVIET parasites. Parasites carrying the CQS (CVMNK) allele displayed a further increase in PIP sensitivity compared with CVIET and SVMNT parasites. Our data indicate that PIP sensitivity was not affected by pfmdr1 sequence status, despite positive correlations between the structurally related compound amodiaquine and pfmdr1 mutations in other studies. In contrast, neither the pfcrt nor pfmdr1 sequence status had any significant impact on susceptibility to dihydroartemisinin. PMID- 19188390 TI - P-glycoprotein-mediated transport of moxifloxacin in a Calu-3 lung epithelial cell model. AB - Moxifloxacin (MXF) is a fluoroquinolone antibiotic that is effective against respiratory infections. However, the mechanisms of MXF lung diffusion are unknown. Active transport in other tissues has been suggested for several members of the fluoroquinolone family. In this study, transport of MXF was systematically investigated across a Calu-3 lung epithelial cell model. MXF showed polarized transport, with the secretory permeability being twice as high as the absorptive permeability. The secretory permeability was concentration dependent (apparent P(max) = 13.6 x 10(-6) cm x s(-1); apparent K(m) = 147 microM), suggesting saturated transport at concentrations higher than 350 microg/ml. The P glycoprotein inhibitor PSC-833 inhibited MXF transport in both directions, whereas probenecid, a multidrug resistance-related protein inhibitor, appeared to have no effect in the Calu-3 model. Moreover, rifampin, a known inducer of efflux transport proteins, upregulated the expression of P-glycoprotein in Calu-3 cells and enhanced MXF active transport. In conclusion, this study clearly indicates that MXF is subject to P-glycoprotein-mediated active transport in the Calu-3 model. This P-glycoprotein-dependent secretion may lead to higher MXF epithelial lining fluid concentrations than those in plasma. Furthermore, drug-drug interactions may be expected when MXF is combined with other P-glycoprotein substrates or modulators. PMID- 19188391 TI - Urinary bactericidal activity of Doripenem versus that of levofloxacin in patients with complicated urinary tract infections or pyelonephritis. AB - The aim of this study was to investigate the urinary bactericidal titers (UBTs) and 24-h area under the UBT-versus-time curve (AUBT) of intravenous doripenem (500 mg every 8 h [q8h]), a new carbapenem, versus those of intravenous levofloxacin (250 mg q24h) in patients with complicated urinary tract infections (cUTIs) or pyelonephritis. UBTs and AUBTs are pharmacokinetic/pharmacodynamic parameters able to reflect the activity of an antimicrobial substance in the urine. Doripenem and levofloxacin show comparable urinary excretion of approximately 80% and are therefore registered for the treatment of UTIs. In order to assess and compare the urinary antimicrobial activities of the two substances, UBTs were investigated for 24 patients (10 treated with doripenem and 14 with levofloxacin) for 31 uropathogens and one control strain. Eight strains were tested for all patients and 27 only in the urine of the corresponding patient. Median UBTs (AUBTs) of doripenem for the uropathogens tested ranged between 1.5 and 65,536 (224 and 909,312) and were significantly higher than median UBTs (AUBTs) of levofloxacin, ranging between 0 and 128 (0 and 2,208). Eight microbiological failures were observed, three after doripenem treatment and five after levofloxacin treatment. For levofloxacin, microbiological failures correlated well with low UBTs and AUBTs, whereas for doripenem there was no correlation. From this study, a calculated target attainment rate for levofloxacin predicting therapeutic success in patients with UTIs approximated mean UBTs of 100 over 24 h or AUBTs of 2,240. Doripenem demonstrated excellent urinary bactericidal activity with the dose administered and appears to be a good alternative in the empirical treatment of cUTI. PMID- 19188392 TI - Pharmacokinetics of hydroxychloroquine and its clinical implications in chemoprophylaxis against malaria caused by Plasmodium vivax. AB - Hydroxychloroquine (HCQ) is an antimalarial drug used as chemoprophylaxis against malaria caused by Plasmodium vivax in the Republic of Korea Army (ROKA). In this study, we evaluated the pharmacokinetics (PK) of HCQ and its metabolites and the relationship between the PK of HCQ and the effect of treatment of HCQ on vivax malaria in South Koreans. Three PK studies of HCQ were conducted with 91 healthy subjects and patients with vivax malaria. Plasma concentrations were analyzed by noncompartmental and mixed-effect modeling approaches. A two-compartment model with first-order absorption best described the data. The clearance and the central and peripheral volumes of distribution were 15.5 liters/h, 733 liters, and 1,630 liters, respectively. We measured the plasma concentrations of HCQ in patients with prophylactic failure of HCQ and compared them with the prediction intervals of the simulated concentrations for HCQ from the final PK model built in this study. In 71% of the patients with prophylactic failure, the plasma concentrations of HCQ were below the lower bounds of the 95% prediction interval, while only 8% of them showed higher levels than the upper bounds of the 95% prediction interval. We report that a significant cause of prophylactic failure among the individuals in ROKA was ascribed to plasma concentrations of HCQ lower than those predicted by the PK model. However, prophylactic failure despite sufficient plasma concentrations of HCQ was confirmed in several individuals, warranting continued surveillance to monitor changes in the HCQ susceptibility of Plasmodium vivax in the Republic of Korea. PMID- 19188393 TI - Intracellular activity of antibiotics in a model of human THP-1 macrophages infected by a Staphylococcus aureus small-colony variant strain isolated from a cystic fibrosis patient: pharmacodynamic evaluation and comparison with isogenic normal-phenotype and revertant strains. AB - Small-colony variant (SCV) strains of Staphylococcus aureus show reduced antibiotic susceptibility and intracellular persistence, potentially explaining therapeutic failures. The activities of oxacillin, fusidic acid, clindamycin, gentamicin, rifampin, vancomycin, linezolid, quinupristin-dalfopristin, daptomycin, tigecycline, moxifloxacin, telavancin, and oritavancin have been examined in THP-1 macrophages infected by a stable thymidine-dependent SCV strain in comparison with normal-phenotype and revertant isogenic strains isolated from the same cystic fibrosis patient. The SCV strain grew slowly extracellularly and intracellularly (1- and 0.2-log CFU increase in 24 h, respectively). In confocal and electron microscopy, SCV and the normal-phenotype bacteria remain confined in acid vacuoles. All antibiotics tested, except tigecycline, caused a net reduction in bacterial counts that was both time and concentration dependent. At an extracellular concentration corresponding to the maximum concentration in human serum (total drug), oritavancin caused a 2-log CFU reduction at 24 h; rifampin, moxifloxacin, and quinupristin-dalfopristin caused a similar reduction at 72 h; and all other antibiotics had only a static effect at 24 h and a 1-log CFU reduction at 72 h. In concentration dependence experiments, response to oritavancin was bimodal (two successive plateaus of -0.4 and -3.1 log CFU); tigecycline, moxifloxacin, and rifampin showed maximal effects of -1.1 to -1.7 log CFU; and the other antibiotics produced results of -0.6 log CFU or less. Addition of thymidine restored intracellular growth of the SCV strain but did not modify the activity of antibiotics (except quinupristin-dalfopristin). All drugs (except tigecycline and oritavancin) showed higher intracellular activity against normal or revertant phenotypes than against SCV strains. The data may help rationalizing the design of further studies with intracellular SCV strains. PMID- 19188394 TI - Population pharmacokinetics of high-dose, prolonged-infusion cefepime in adult critically ill patients with ventilator-associated pneumonia. AB - A population pharmacokinetic model of cefepime was constructed from data from adult critical care patients with ventilator-associated pneumonia (VAP). A total of 32 patients treated with high-dose cefepime, 2 g every 8 h (3-h infusion) or a renal function-adjusted equivalent dose, were randomized into two groups--26 for the initial model and 6 for model validation. Serum samples of cefepime were collected at steady state. Nonparametric adaptive grid population modeling was employed using a two-compartment K(slope) pharmacokinetic model relating the elimination rate constant (K(10)) to renal function, as defined by creatinine clearance (CL(CR)), and central distribution volume (V(1)) to total body weight (TBW). The final model was described by the following equations: K(10) = 0.0027 x CL(CR) + 0.071 h(-1) and V(1) = TBW x 0.21 liter/kg. The median intercompartmental transfer constants K(12) and K(21) were 0.780 h(-1) and 0.472 h(-1), respectively. Using these median parameter estimates, the bias, precision, and coefficient of determination for the initial model were 11.3 microg/ml, 24.0 microg/ml, and 26%, respectively. The independent validation group displayed a bias, precision, and coefficient of determination of -1.64 microg/ml, 17.1 microg/ml, and 62%, respectively. Time-concentration profiles were assessed for various dosing regimens, using 5,000-patient Monte Carlo simulations. Among the regimens, the likelihoods of 2 g every 8 h (3-h infusion) achieving free drug concentrations above the MIC for 50% of the dosing interval were 91.8%, 78.1%, and 50.3% for MICs of 8, 16, and 32 microg/ml, respectively. This study provides a pharmacokinetic model capable of predicting cefepime concentrations in critically ill patients with VAP. PMID- 19188395 TI - High cerebrospinal fluid (CSF) penetration and potent bactericidal activity in CSF of NZ2114, a novel plectasin variant, during experimental pneumococcal meningitis. AB - Plectasin is the first defensin-type antimicrobial peptide isolated from a fungus and has potent activity against gram-positive bacteria. By using an experimental meningitis model, the penetration of plectasin into the cerebrospinal fluid (CSF) of infected and uninfected rabbits and the bactericidal activities in CSF of the plectasin variant NZ2114 and ceftriaxone against a penicillin-resistant Streptococcus pneumoniae strain (NZ2114 and ceftriaxone MICs, 0.25 and 0.5 microg/ml, respectively) were studied. Pharmacokinetic analysis showed that there was a significantly higher level of CSF penetration of NZ2114 through inflamed than through noninflamed meninges (area under the concentration-time curve for CSF/area under the concentration-time curve for serum, 33% and 1.1%, respectively; P = 0.03). The peak concentrations of NZ2114 in purulent CSF were observed approximately 3 h after the infusion of an intravenous bolus of either 20 or 40 mg/kg of body weight and exceeded the MIC >10-fold for a 6-h study period. Treatment with NZ2114 (40 and 20 mg/kg at 0 and 5 h, respectively; n = 11) caused a significantly higher reduction in CSF bacterial concentrations than therapy with ceftriaxone (125 mg/kg at 0 h; n = 7) at 3 h (median changes, 3.7 log(10) CFU/ml [interquartile range, 2.5 to 4.6 log(10) CFU/ml] and 2.1 log(10) CFU/ml [interquartile range, 1.7 to 2.6 log(10) CFU/ml], respectively; P = 0.001), 5 h (median changes, 5.2 log(10) CFU/ml [interquartile range, 3.6 to 6.1 log(10) CFU/ml] and 3.1 log(10) CFU/ml [interquartile range, 2.6 to 3.7 log(10) CFU/ml], respectively; P = 0.01), and 10 h (median changes, 5.6 log(10) CFU/ml [interquartile range, 5.2 to 5.9 log(10) CFU/ml] and 4.2 log(10) CFU/ml [interquartile range, 3.6 to 5.0 log(10) CFU/ml], respectively; P = 0.03) after the start of therapy as well compared to the CSF bacterial concentrations in untreated rabbits with meningitis (n = 7, P < 0.05). Also, significantly more rabbits had sterile CSF at 5 and 10 h when they were treated with NZ2114 than when they were treated with ceftriaxone (67% [six of nine rabbits] and 0% [zero of seven rabbits], respectively, at 5 h and 75% [six of eight rabbits] and 14% [one of seven rabbits], respectively, at 10 h; P < 0.05). Due to its excellent CSF penetration and potent bactericidal activity in CSF, the plectasin variant NZ2114 could be a promising new option for the treatment of CNS infections caused by gram-positive bacteria, including penicillin-resistant pneumococcal meningitis. PMID- 19188396 TI - Antibiotic susceptibility and characteristics of Neisseria meningitidis isolates from the African meningitis belt, 2000 to 2006: phenotypic and genotypic perspectives. AB - Up-to-date information regarding the antibiotic susceptibility of Neisseria meningitidis strains from African countries is highly limited. Our aim was to comprehensively describe the antibiotic susceptibilities of a selection of N. meningitidis isolates recovered between 2000 and 2006 from 18 African countries, mainly those within the meningitis belt. Susceptibilities to 11 antibiotics were determined using Etest for 137 N. meningitidis isolates (stringently selected from 693 available isolates). The isolates were also characterized by serogrouping, multilocus sequence typing, genosubtyping, and penA allele identification. All N. meningitidis isolates were susceptible to ceftriaxone, chloramphenicol, and ciprofloxacin. No isolate produced beta-lactamase. Only three isolates (2%) displayed reduced susceptibility to penicillin G. The two isolates with the highest penicillin G MICs were the only isolates showing reduced susceptibility to ampicillin and cefuroxime. One of these isolates was also resistant to penicillin V. One percent of isolates displayed reduced susceptibility to rifampin, while 52% of the isolates were resistant to tetracycline, 74% were resistant to erythromycin, and 94% were resistant to sulfadiazine. The MICs of rifampin and tetracycline seemed to be associated with the serogroup of the isolates. In total, 18 sequence types (STs), 10 genosubtypes, and 8 different penA alleles were identified; the most common were ST-7, P1.20,9,35-1, and penA4, respectively. A high level of correlation was found between ST, genosubtype, and penA allele. In conclusion, N. meningitidis isolates from the African meningitis belt remain highly susceptible to the antibiotics used. Regarding beta-lactam antibiotics, rare isolates showed a reduced susceptibility to penicillins, but the expanded-spectrum cephalosporins are not affected at present. PMID- 19188397 TI - Intracellular activity of antibiotics in a model of human THP-1 macrophages infected by a Staphylococcus aureus small-colony variant strain isolated from a cystic fibrosis patient: study of antibiotic combinations. AB - In a companion paper (H. A. Nguyen et al., Antimicrob. Agents Chemother. 53:1434 1442, 2009), we showed that vancomycin, oxacillin, fusidic acid, clindamycin, linezolid, and daptomycin are poorly active against the intracellular form of a thymidine-dependent small-colony variant (SCV) strain isolated from a cystic fibrosis patient and that the activity of quinupristin-dalfopristin, moxifloxacin, rifampin, and oritavancin remains limited (2- to 3-log CFU reduction) compared to their extracellular activity. Antibiotic combination is a well-known strategy to improve antibacterial activity, which was examined here against an intracellular SCV strain using combinations with either rifampin or oritavancin. Time-kill curve analysis using either concentrations that caused a static effect for each antibiotic individually or concentrations corresponding to the maximum concentration in human serum showed largely divergent effects that were favorable when antibiotics were combined with rifampin at low concentrations only and with oritavancin at both low and high concentrations. The nature of the interaction between rifampin, oritavancin, and moxifloxacin was further examined using the fractional maximal effect method, which allows categorization of the effects of combinations when dose-effect relationships are not linear. Rifampin and oritavancin were synergistic at all concentration ratios investigated. Oritavancin and moxifloxacin were also synergistic but at high oritavancin concentrations only. Rifampin and moxifloxacin were additive. This approach may help in better assessing and improving the activity of antibiotics against intracellular SCV strains. PMID- 19188398 TI - Surveillance of carbapenem-resistant Pseudomonas aeruginosa isolates from Puerto Rican Medical Center Hospitals: dissemination of KPC and IMP-18 beta-lactamases. AB - During a 6-month period, 37/513 (7.2%) Pseudomonas aeruginosa isolates belonging to 13 pulsed-field gel electrophoresis (PFGE) groups from Puerto Rican hospitals were carbapenem nonsusceptible. Seven of 37 isolates from four PFGE groups carried bla(IMP-18), and 25/37 isolates from seven PFGE groups carried bla(KPC). The results indicated the clonal spread of bla(KPC)-positive P. aeruginosa isolates into several Puerto Rican hospitals and the dissemination of bla(IMP-18) and bla(KPC) into genetically unrelated isolates. PMID- 19188400 TI - A home away from home: the patient-centered health home. PMID- 19188399 TI - New high-throughput screening assay to reveal similarities and differences in inhibitory sensitivities of multidrug ATP-binding cassette transporters. AB - Cdr1p is the major ATP-binding cassette multidrug transporter conferring resistance to azoles and other antifungals in Candida albicans. In this study, the identification of new Cdr1p inhibitors by use of a newly developed high throughput fluorescence-based assay is reported. The assay also allowed monitoring of the activity and inhibition of the related transporters Pdr5p and Snq2p of Saccharomyces cerevisiae, which made it possible to compare its performance with those of previously established procedures. A high sensitivity, resulting from a wide dynamic range, was achieved upon high-level expression of the Cdr1p, Pdr5p, and Snq2p transporters in an S. cerevisiae strain in which the endogenous interfering activities were further reduced by genetic manipulation. An analysis of a set of therapeutically used and newly synthesized phenothiazine derivatives revealed different pharmacological profiles for Cdr1p, Pdr5p, and Snq2p. All transporters showed similar sensitivities to M961 inhibition. In contrast, Cdr1p was less sensitive to inhibition by fluphenazine, whereas phenothiazine selectively inhibited Snq2p. The inhibition potencies measured by the new assay reflected the ability of the compounds to potentiate the antifungal effect of ketoconazole (KTC), which was detoxified by the overproduced transporters. They also correlated with the 50% inhibitory concentration for inhibition of Pdr5p-mediated transport of rhodamine 6G in isolated plasma membranes. The most active derivative, M961, potentiated the activity of KTC against an azole-resistant CDR1-overexpressing C. albicans isolate. PMID- 19188401 TI - Gingival retraction. PMID- 19188402 TI - Inspired. PMID- 19188403 TI - A second opinion. PMID- 19188404 TI - 'Periodontal maintenance'. PMID- 19188405 TI - Quitting tobacco use. PMID- 19188406 TI - Biology of esthetics. PMID- 19188407 TI - Full-face images. PMID- 19188408 TI - Rating smiles. PMID- 19188409 TI - Study finds link between oral inflammatory diseases and Epstein-Barr virus. PMID- 19188410 TI - Gene therapy stops periodontal disease development. PMID- 19188411 TI - Comorbidities affect survival of patients with head, neck cancer. PMID- 19188412 TI - A randomized trial of resin-based restorations in class I and class II beveled preparations in primary molars: 24-month results. AB - PURPOSE: The authors conducted a randomized clinical trial to evaluate the survival rate of esthetic restorations in Class I and Class II beveled preparations in primary molars 24 months after placement. The null hypothesis was that there is no difference among survival rates of the restorative materials used. METHODS: Forty-eight children (mean age, 5 years 9 months) received 141 restorations in beveled cavosurface margins in primary molars randomly assigned by lottery method: 46 received treatment with Vitremer Tri-Cure Glass Ionomer System (3M ESPE Dental Products, St. Paul, Minn.) (33 Class I and 13 Class II restorations), 51 received treatment with Freedom (SDI, Bayswater, Victoria, Australia) (36 Class I and 15 Class II restorations); 44 received treatment with TPH Spectrum (Dentsply, Petropolis, Rio de Janeiro, Brazil) (30 Class I and 14 Class II restorations). Two examiners whose technique had been calibrated (weight kappa > 0.85) evaluated the restorations using modified U.S. Public Health Service criteria and Visible Plaque Index score at baseline and at 12, 18 and 24 months. RESULTS: After two years, the authors censored data for 17 restorations, considered 101 restorations to be clinically successful and deemed 23 restorations failed because of loss of marginal integrity, anatomical form discrepancies and secondary caries. For Class I and Class II restorations, the cumulative survival rates were higher than 80 percent and 55 percent, respectively, for all materials (life table, Gehan-Wilcoxon Test, P > .05; P > .05). CONCLUSIONS: At the 24-month clinical recall, the authors found no differences among materials in Class I (P > .05) or Class II beveled preparations (P > .05) in primary molars, but all materials showed higher survival rates in Class I than in Class II restorations. PMID- 19188413 TI - Atrial fibrillation: pathogenesis, medical-surgical management and dental implications. AB - BACKGROUND: Atrial fibrillation (AF) is a cardiac rhythm disturbance arising from disorganized electrical activity in the atria, and it is accompanied by an irregular and often rapid ventricular response. It is the most common clinically significant dysrhythmia in the general and older population. TYPES OF STUDIES REVIEWED: The authors conducted a MEDLINE search using the key terms "atrial fibrillation," "epidemiology," "pathophysiology," "treatment" and "dentistry." They selected contemporaneous articles published in peer-reviewed journals and gave preference to articles reporting randomized controlled trials. CLINICAL IMPLICATIONS: The anticoagulant warfarin frequently is prescribed to prevent stroke caused by cardiogenic thromboemboli arising from stagnant blood in poorly contracting atria. Most dental procedures and a limited number of surgical procedures can be performed without altering warfarin dosage if the international normalized ratio value is within the therapeutic range of 2.0 to 3.0. Certain analgesic agents, antibiotic agents, antifungal agents and sedative hypnotics, however, should not be prescribed without consultation with the patient's physician because these medications may alter the patient's risk of hemorrhage and stroke. CONCLUSIONS: AF affects nearly 2.5 million Americans, most of who are older than 60 years. Consultation with the patient's physician to discuss the planned dental treatment often is appropriate, especially for people who frequently have comorbid diseases such as coronary artery disease, congestive heart failure, diabetes and thyrotoxicosis, which are treated with multiple drug regimens. PMID- 19188414 TI - Potential mechanisms underpinning the nutritional modulation of periodontal inflammation. AB - BACKGROUND: Periodontitis results from an inappropriate host response to pathogenic biofilms. Because traditional management approaches have failed to reduce disease prevalence, the research focus has shifted toward managing host mediated inflammation. In this article, the author reviews the role of nutrition in the development and resolution of inflammation. METHODS: The author reviewed the biomedical literature to elucidate mechanisms by which dietary factors affect inflammatory processes and to establish what evidence exists for macronutritional and micronutritional modulation of inflammation at a cellular and molecular level. CONCLUSIONS: Hyperinflammation characterizes the periodontitis phenotype, and oxidative stress is a key orchestration point for the diverse signaling pathways, which control inflammation. Oxidative stress is modulated by diet, as well as by infection. Recent research has demonstrated that subtle shifts in nutritional status are associated independently with the prevalence of periodontitis. Moreover, the results of contemporary animal and human studies have demonstrated the role of specific micronutrients in the modulation of the host's inflammatory response by reducing inflammatory biomarkers and bone loss. CLINICAL IMPLICATIONS: The scientific community is starting to realize the health benefits of diets containing foods naturally rich in antioxidants and omega-3 polyunsaturated fatty acids, as well as the dangers of diets that are high in refined carbohydrates. Nutritional intervention studies in patients with inflammatory periodontitis are needed to evaluate the effect of nutritional approaches to periodontal management. PMID- 19188415 TI - Asymptomatic pebbly lesions. PMID- 19188416 TI - Evaluation of two nitrous oxide scavenging systems using infrared thermography to visualize and control emissions. AB - BACKGROUND: The authors conducted a study to determine the effectiveness of two waste anesthetic gas-scavenging systems. They also evaluated one of the systems to determine the effect of work practices in controlling waste nitrous oxide (N2O). METHODS: The authors collected a minimum of 13 data sets in each phase of the study that included infrared thermography, digital videography and real-time air analysis for ambient concentrations of waste N2O. Surgeon 1, who had experience using both systems, used the Safe Sedate Dental Mask (Airgas, Radnor, Pa.) system (system I) in phase I and the Porter Nitrous Oxide Sedation System (Porter Instruments, Hatfield, Pa.) (system II) in phase II. Surgeon 2, who did not have experience using system I, used it in phase III. To evaluate each system's effectiveness, the authors collected N2O air concentration data from phases I and II and compared the data with the National Institute for Occupational Safety and Health Recommended Exposure Limit (NIOSH REL). They also compared phases I and III to determine the effect of work practices on the systems' effectiveness. RESULTS: Surgeon 1 controlled occupational exposure to N2O significantly better using system I than using system II. Mean N2O air concentration levels during phases I and II were 61.6 parts per million (ppm) and 225.6 ppm, respectively. Surgeon 2 did not achieve results comparable to those of surgeon 1 in phase I using system I. Infrared thermography and air concentration data suggested that key work practices and patient and surgical variables accounted for the different results obtained in phases I and III. CONCLUSIONS: Although neither system was able to control occupational exposure of N2O oxide below the NIOSH REL, system I met the American Conference of Governmental Industrial Hygienists threshold limit value of less than 50 ppm during an eight hour day and performed significantly better than did system II. CLINICAL IMPLICATIONS: System I achieved maximal efficiency when combined with consistent best work practices. PMID- 19188417 TI - An evaluation of replacement rates for posterior resin-based composite and amalgam restorations in U.S. Navy and marine corps recruits. AB - BACKGROUND: Restoration replacement is a clinical concern that has not been studied among military personnel. The authors determined the prevalence of placement of posterior amalgam and resin-based composite restorations and the incidence of replacement among U.S. Navy and Marine Corps personnel. METHODS: The authors analyzed dental records from 2,780 personnel to determine the relative risk of replacement for initially sound restorations during subjects' first years of military service. RESULTS: At the initial examination, 964 (15.2 percent) of amalgam restorations and 199 (17.4 percent) of resin-based composite restorations required re-treatment. Of those judged clinically acceptable, 14.2 percent of amalgam and 16.7 percent of resin-based composite restorations required replacement during the observation period. The authors found significant increases in replacement rates for resin-based composite restorations compared with amalgam restorations for replacement due to all causes (adjusted hazard ratio, 1.28; P < .05), as well as for replacement due to restoration failure (adjusted hazard ratio, 1.64; P < .01). CONCLUSIONS: About 30 percent of posterior restorations required replacement, either at the initial examination or during the subjects' first years of military service. In a young military population, significantly more resin-based composite restorations in place at the initial examination will require replacement than will amalgam restorations. Multi-surface restorations had higher rates of replacement than did one-surface restorations, and subjects at high caries risk experienced significantly higher replacement rates than did those at low caries risk. CLINICAL IMPLICATIONS: The number of surfaces restored and subjects' caries risk status may influence the longevity of resin-based composite and amalgam restorations. PMID- 19188418 TI - Providing dental care to pregnant patients: a survey of Oregon general dentists. AB - BACKGROUND: A growing number of studies and reports indicate preventive, routine and emergency dental procedures can be provided safely to pregnant patients to alleviate dental problems and promote oral health of mothers and children. METHODS: In 2006 and 2007, the authors conducted a survey of 1,604 general dentists in Oregon. The survey asked dentists about their attitudes, beliefs and practices regarding dental care for pregnant patients. The authors compared the responses with 2006 guidelines from a New York State Department of Health expert panel. RESULTS: The response rate was 55.2 percent. Most respondents (91.7 percent) agreed that dental treatment should be part of prenatal care. Two-thirds of respondents (67.7 percent) were interested in receiving continuing dental education (CDE) regarding the care of pregnant patients. Comparisons of self reported knowledge and practice with the aforementioned guidelines revealed several points of difference; the greatest regarded obtaining full-mouth radiographs, providing nitrous oxide, administering long-acting anesthetic injections and use of over-the-counter pain medications. CONCLUSIONS: Dentists need pregnancy-specific education to provide up-to-date preventive and curative care to pregnant patients. The results of the study identified specific skills and misinformation that could be addressed through CDE. CLINICAL IMPLICATIONS: Comprehensive dental care provided during pregnancy is needed to ensure the oral health of all women at risk of experiencing pregnancy-specific problems, as well as the prevention of early childhood caries. PMID- 19188419 TI - Clinical characteristics and diagnosis of atypical odontalgia: implications for dentists. AB - BACKGROUND: Atypical odontalgia (AO) is a poorly understood and commonly misdiagnosed condition for which patients often undergo multiple unsuccessful dental or surgical procedures. The authors conducted a study to determine the prevalence and describe the characteristics of patients with AO seen at the University of Southern California Orofacial Pain and Oral Medicine Center (USC OFP-OM Center), Los Angeles. METHODS: The authors conducted a retrospective record review from a database of more than 3,000 patient records from June 2003 to August 2007 to identify patients diagnosed with AO. RESULTS: The authors identified 64 patients (44 women and 20 men) between the ages of 26 and 93 years as having a diagnosis of AO. Of those 64 patients, 71 percent initially consulted a dentist regarding their pain, and 79 percent had undergone dental treatment that failed to resolve the pain. The pain of 64 percent of the patients had no known cause. CONCLUSIONS: Dentists, who often are the first health care providers to see patients with AO, must be aware of this condition and must follow the appropriate steps to determine its diagnosis. CLINICAL IMPLICATIONS: Dentists and physicians should understand the implications and importance of early diagnosis of patients with AO and of referral to pain specialists for treatment. PMID- 19188420 TI - The development and testing of a survey instrument for benchmarking dental plan performance: using insured patients' experiences as a gauge of dental care quality. AB - BACKGROUND: There is no standard, nonproprietary method for providing national benchmarks of dental care quality as described by patients. The purpose of this research was to develop such a tool following guidelines of the Consumer Assessment of Healthcare Providers and Systems (CAHPS) initiative. METHODS: The authors identified domains of dental care quality via qualitative methods, including a literature review, stakeholder interviews and focus groups with beneficiaries, and they cognitively tested draft questions with patients to yield a pilot survey. Psychometric analyses of pilot data (n = 3,264) identified summary indexes and guided survey revisions. The authors used two waves of subsequent data collection (n = 4,221) to test the validity of the revised survey. RESULTS: The mean response rate across three rounds of data collection was 51 percent. Statistical analysis indicated that 17 questions could be reliably collapsed into three composite measures: "Care From Dentist and Staff" (reliability = 0.89, scaling success = 100 percent); "Access to Dental Care" (reliability = 0.78, scaling success = 100 percent); and "Dental Plan Coverage/Service" (reliability = 0.84, scaling success = 100 percent). CONCLUSIONS: The validity of the survey was supported in mail and Internet modes for the American English language, and the instrument was approved by the CAHPS consortium for distribution as the CAHPS Dental Plan Survey. PRACTICE IMPLICATIONS. A tool is available now for assessing dental care quality by measuring adult patients' experiences with their dental care and coverage. The authors tested this instrument only in a population with third-party coverage, however, which is a potential limitation that should be considered. PMID- 19188421 TI - Restorative dentistry for times of economic distress. PMID- 19188422 TI - Understanding the practice management component of dental implants. PMID- 19188423 TI - What are the ethical issues I should consider when contemplating exclusive territory agreements? PMID- 19188424 TI - Munc18c depletion selectively impairs the sustained phase of insulin release. AB - OBJECTIVE: The Sec1/Munc18 protein Munc18c has been implicated in Syntaxin 4 mediated exocytosis events, although its purpose in exocytosis has remained elusive. Given that Syntaxin 4 functions in the second phase of glucose stimulated insulin secretion (GSIS), we hypothesized that Munc18c would also be required and sought insight into the possible mechanism(s) using the islet beta cell as a model system. RESEARCH DESIGN AND METHODS: Perifusion analyses of isolated Munc18c- (-/+) or Munc18c-depleted (RNAi) mouse islets were used to assess biphasic secretion. Protein interaction studies used subcellular fractions and detergent lysates prepared from MIN6 beta-cells to determine the mechanistic role of Munc18c in Syntaxin 4 activation and docking/fusion of vesicle-associated membrane protein (VAMP)2-containing insulin granules. Electron microscopy was used to gauge changes in granule localization. RESULTS: Munc18c (-/+) islets secreted approximately 60% less insulin selectively during second-phase GSIS; RNAi-mediated Munc18c depletion functionally recapitulated this in wild-type and Munc18c (-/+) islets in a gene dosage-dependent manner. Munc18c depletion ablated the glucose-stimulated VAMP2-Syntaxin 4 association as well as Syntaxin 4 activation, correlating with the deficit in insulin release. Remarkably, Munc18c depletion resulted in aberrant granule localization to the plasma membrane in response to glucose stimulation, consistent with its selective effect on the second phase of secretion. CONCLUSIONS: Collectively, these studies demonstrate an essential positive role for Munc18c in second-phase GSIS and suggest novel roles for Munc18c in granule localization to the plasma membrane as well as in triggering Syntaxin 4 accessibility to VAMP2 at a step preceding vesicle docking/fusion. PMID- 19188425 TI - MicroRNAs induced during adipogenesis that accelerate fat cell development are downregulated in obesity. AB - OBJECTIVE: We investigated the regulation and involvement of microRNAs (miRNAs) in fat cell development and obesity. RESEARCH DESIGN AND METHODS: Using miRNA microarrays, we profiled the expression of >370 miRNAs during adipogenesis of preadipocyte 3T3-L1 cells and adipocytes from leptin deficient ob/ob and diet induced obese mice. Changes in key miRNAs were validated by RT-PCR. We further assessed the contribution of the chronic inflammatory environment in obese adipose tissue to the dysregulated miRNA expression by tumor necrosis factor (TNF)-alpha treatment of adipocytes. We functionally characterized two adipocyte enriched miRNAs, miR-103 and miR-143, by a gain-of-function approach. RESULTS: Similar miRNAs were differentially regulated during in vitro and in vivo adipogenesis. Importantly, miRNAs that were induced during adipogenesis were downregulated in adipocytes from both types of obese mice and vice versa. These changes are likely associated with the chronic inflammatory environment, since they were mimicked by TNF-alpha treatment of differentiated adipocytes. Ectopic expression of miR-103 or miR-143 in preadipocytes accelerated adipogenesis, as measured both by the upregulation of many adipogenesis markers and by an increase in triglyceride accumulation at an early stage of adipogenesis. CONCLUSIONS: Our results provide the first experimental evidence for miR-103 function in adipose biology. The remarkable inverse regulatory pattern for many miRNAs during adipogenesis and obesity has important implications for understanding adipose tissue dysfunction in obese mice and humans and the link between chronic inflammation and obesity with insulin resistance. PMID- 19188426 TI - Specific local cardiovascular changes of Nepsilon-(carboxymethyl)lysine, vascular endothelial growth factor, and Smad2 in the developing embryos coincide with maternal diabetes-induced congenital heart defects. AB - OBJECTIVE: Embryos exposed to a diabetic environment in utero have an increased risk to develop congenital heart malformations. The mechanism behind the teratogenicity of diabetes still remains enigmatic. Detrimental effects of glycation products in diabetic patients have been well documented. We therefore studied a possible link between glycation products and the development of congenital cardiovascular malformations. Furthermore, we investigated other possible mechanisms involved in this pathogenesis: alterations in the levels of vascular endothelial growth factor (VEGF) or phosphorylated Smad2 (the latter can be induced by both glycation products and VEGF). RESEARCH DESIGN AND METHODS: We examined the temporal spatial patterning of the glycation products Nepsilon(carboxymethyl)lysine (CML) and methylglyoxal (MG) adducts, VEGF expression, and phosphorylated Smad2 during cardiovascular development in embryos from normal and diabetic rats. RESULTS: Maternal diabetes increased the CML accumulation in the areas susceptible to diabetes-induced congenital heart disease, including the outflow tract of the heart and the aortic arch. No MG adducts could be detected, suggesting that CML is more likely to be indicative for increased oxidative stress than for glycation. An increase of CML in the outflow tract of the heart was accompanied by an increase in phosphorylated Smad2, unrelated to VEGF. VEGF showed a time-specific decrease in the outflow tract of embryos from diabetic dams. CONCLUSIONS: From our results, we can conclude that maternal diabetes results in transient and localized alterations in CML, VEGF expression, and Smad2 phosphorylation overlapping with those regions of the developing heart that are most sensitive to diabetes-induced congenital heart disease. PMID- 19188427 TI - Interleukin-6 attenuates insulin-mediated increases in endothelial cell signaling but augments skeletal muscle insulin action via differential effects on tumor necrosis factor-alpha expression. AB - OBJECTIVE: The cytokine interleukin-6 (IL-6) stimulates AMP-activated protein kinase (AMPK) and insulin signaling in skeletal muscle, both of which result in the activation of endothelial nitric oxide synthase (eNOS). We hypothesized that IL-6 promotes endothelial cell signaling and capillary recruitment in vivo, contributing to increased glucose uptake. RESEARCH DESIGN AND METHODS: The effect of IL-6 with and without insulin on AMPK, insulin, and eNOS signaling in and nitric oxide (NO) release from human aortic endothelial cells (HAECs) was examined. The physiological significance of these in vitro signaling events was assessed by measuring capillary recruitment in rats during control and euglycemic hyperinsulinemic clamps with or without IL-6 infusion. RESULTS: IL-6 blunted increases in insulin signaling, eNOS phosphorylation (Ser1177), and NO production and reduced phosphorylation of AMPK in HAEC in vitro and capillary recruitment in vivo. In contrast, IL-6 increased Akt phosphorylation (Ser473) in hindlimb skeletal muscle and enhanced whole-body glucose disappearance and glucose uptake during the clamp. The differences in endothelial cell and skeletal muscle signaling were mediated by the cell-specific, additive effects of IL-6 and insulin because this treatment markedly increased tumor necrosis factor (TNF) alpha protein expression in HAECs without any effect on TNF-alpha in skeletal muscle. When HAECs were incubated with a TNF-alpha-neutralizing antibody, the negative effects of IL-6 on eNOS signaling were abolished. CONCLUSIONS: In the presence of insulin, IL-6 contributes to aberrant endothelial cell signaling because of increased TNF-alpha expression. PMID- 19188428 TI - Weight loss may reverse blunted sympathetic neural responsiveness to glucose ingestion in obese subjects with metabolic syndrome. AB - OBJECTIVE: The purpose of this study was to examine the effects of weight loss on sympathetic nervous system responsiveness to glucose ingestion in obese subjects with metabolic syndrome, in whom such responses are reportedly blunted. RESEARCH DESIGN AND METHODS: Thirty four subjects, 19 insulin resistant and 15 insulin sensitive and aged 55 +/- 1 years (mean +/- SE) with BMI 31.6 +/- 0.6 kg/m2, who fulfilled the Adult Treatment Panel III criteria for metabolic syndrome participated. Simultaneous measurements of whole-body norepinephrine spillover rate, calf blood flow, and intra-arterial blood pressure were made at times 0, 30, 60, 90, and 120 min postglucose (75 g). The experiment was repeated after a 3 month hypocaloric diet with or without an exercise program. RESULTS: Body weight decreased by 8.1 +/- 0.9 and 8.4 +/- 1.1 kg and resting norepinephrine spillover by 94 +/- 31 and 166 +/- 58 ng/min (all P < or = 0.01) in insulin-resistant and insulin-sensitive subjects, respectively. Weight loss was accompanied by a marked increase in sympathetic responsiveness after glucose but only in insulin resistant subjects. In this subgroup, comparative increases in norepinephrine spillover rates at baseline and after weight loss averaged -3 +/- 25 versus 73 +/ 24 ng/min at 30 min (P = 0.039), 36 +/- 21 versus 115 +/- 28 ng/min at 60 min (P = 0.045), 9 +/- 21 versus 179 +/- 50 ng/min at 90 min (P < 0.001), and 40 +/- 48 versus 106 +/- 39 ng/min at 120 min (P = 0.24). CONCLUSIONS: Weight loss reverses blunted sympathetic responsiveness to glucose ingestion in insulin-resistant subjects with metabolic syndrome, which is relevant to postprandial energy utilization and body weight homeostasis. PMID- 19188429 TI - Protective effect of perindopril on diabetic retinopathy is associated with decreased vascular endothelial growth factor-to-pigment epithelium-derived factor ratio: involvement of a mitochondria-reactive oxygen species pathway. AB - OBJECTIVE: This study aimed to verify whether the decreased vascular endothelial growth factor (VEGF)-to-pigment epithelium-derived factor (PEDF) ratio can serve as an indicator for the protective effect of angiotensin-converting enzyme inhibitors (ACEIs) on diabetic retinopathy (DR) and to investigate the role of mitochondrial reactive oxygen species (ROS) in the downregulated VEGF-to-PEDF ratio. RESEARCH DESIGN AND METHODS: Diabetic rats and control animals were randomly assigned to receive perindopril or vehicle for 24 weeks, and bovine retinal capillary endothelial cells (BRECs) were incubated with normal or high glucose with or without perindopril. VEGF, PEDF, PPARgamma, and uncoupling protein-2 (UCP-2) in the rat retinas or BREC extracts were examined by Western blotting and real-time RT-PCR. The levels of VEGF and PEDF in cell culture media were examined by ELISA. Mitochondrial membrane potential (Deltapsim) and ROS production were assayed using JC-1 or CM-H2DCFDA. RESULTS: The VEGF-to-PEDF ratio was increased in the retina of diabetic rats; perindopril lowered the increased VEGF-to-PEDF ratio in diabetic rats and ameliorated the retinal damage. In BRECs, perindopril lowered the hyperglycemia-induced elevation of VEGF-to-PEDF ratio by reducing mitochondrial ROS. We found the decreased ROS production was a result of perindopril-induced upregulation of PPARgamma and UCP-2 expression and the subsequent decrease of Deltapsim. CONCLUSIONS: It is concluded that the protective effect of ACEI on DR is associated with a decreased VEGF-to-PEDF ratio, which involves the mitochondria-ROS pathway through PPARgamma-mediated changes of UCP-2. This study paves a way for future application of ACEI in treatment of DR. PMID- 19188430 TI - Liver-specific loss of lipolysis-stimulated lipoprotein receptor triggers systemic hyperlipidemia in mice. AB - OBJECTIVE: In mammals, proper storage and distribution of lipids in and between tissues is essential for the maintenance of energy homeostasis. In contrast, aberrantly high levels of triglycerides in the blood ("hypertriglyceridemia") represent a hallmark of the metabolic syndrome and type 2 diabetes. As hypertriglyceridemia has been identified as an important risk factor for cardiovascular complications, in this study we aimed to identify molecular mechanisms in aberrant triglyceride elevation under these conditions. RESEARCH DESIGN AND METHODS: To determine the importance of hepatic lipid handling for systemic dyslipidemia, we profiled the expression patterns of various hepatic lipid transporters and receptors under healthy and type 2 diabetic conditions. A differentially expressed lipoprotein receptor was functionally characterized by generating acute, liver-specific loss- and gain-of-function animal models. RESULTS: We show that the hepatic expression of lipid transporter lipolysis stimulated lipoprotein receptor (LSR) is specifically impaired in mouse models of obesity and type 2 diabetes and can be restored by leptin replacement. Experimental imitation of this pathophysiological situation by liver-specific knockdown of LSR promotes hypertriglyceridemia and elevated apolipoprotein (Apo)B and E serum levels in lean wild-type and ApoE knockout mice. In contrast, genetic restoration of LSR expression in obese animals to wild-type levels improves serum triglyceride levels and serum profiles in these mice. CONCLUSIONS: The dysregulation of hepatic LSR under obese and diabetic conditions may provide a molecular rationale for systemic dyslipidemia in type 2 diabetes and the metabolic syndrome and represent a novel target for alternative treatment strategies in these patients. PMID- 19188431 TI - Prenatal stress or high-fat diet increases susceptibility to diet-induced obesity in rat offspring. AB - OBJECTIVE: Perturbations to the prenatal environment have been associated with the development of adult chronic disease, findings that gave rise to the "Barker Hypothesis" or the "developmental origins of adult disease" concept. In this study, we used an animal model to determine the metabolic consequences of maternal prenatal stress and high-fat feeding on the developing offspring. RESEARCH DESIGN AND METHODS: Pregnant female Sprague-Dawley rats were maintained on standard chow or 60% high-fat diet throughout gestation and lactation. Half of each group were exposed to a novel variable stress paradigm during the 3rd week of gestation, whereas control dams were left undisturbed. Body weight, body composition, glucose tolerance, and endocrine parameters were measured in offspring through early adulthood. RESULTS: Male and female pups from dams that experienced prenatal stress and/or were on a high-fat diet weighed more beginning on postnatal day 7 compared with standard chow-control pups. Access to high-fat diet at weaning increased the body weight effect through early adulthood and was attributable to greater adiposity. Pups weaned onto standard chow diet showed no significant difference in glucose clearance or insulin secretion. However, pups weaned onto high-fat diet had impaired glucose tolerance if their dams were on a high-fat diet, experienced prenatal stress, or both. CONCLUSIONS: Our data demonstrate that prenatal stress and/or high-fat diet during the intrauterine or postnatal environment affects offspring in a manner that increases their susceptibility to diet-induced obesity and leads to secondary adverse metabolic consequences. PMID- 19188432 TI - Breast-feeding modifies the association of PPARgamma2 polymorphism Pro12Ala with growth in early life: the Generation R Study. AB - OBJECTIVE: We examined whether the PPARgamma2 Ala12 allele influences growth in early life and whether this association is modified by breast-feeding. RESEARCH DESIGN AND METHODS: This study was embedded in the Generation R Study, a prospective cohort study from early fetal life onward. PPARgamma2 was genotyped in DNA obtained from cord blood samples in 3,432 children. Information about breast-feeding was available from questionnaires. Weight, head circumference, and femur length were repeatedly measured in second and third trimesters of pregnancy, at birth, and at the ages of 1.5, 6, 11, 14, and 18 months. RESULTS: Genotype frequency distribution was 77.6% (Pro12Pro), 20.7% (Pro12Ala), and 1.7% (Ala12Ala). Growth rates in weight from second trimester of pregnancy to 18 months were higher for Pro12Ala and Ala12Ala than for Pro12Pro carriers (differences 1.11 g/week [95% CI 0.47-1.74] and 2.65 g/week [0.45-4.87], respectively). We found an interaction between genotype and breast-feeding duration (P value for interaction <0.0001). In infants who were breast-fed for > or =4 months, PPARgamma2 Pro12Ala was not associated with growth rate. When breast-feeding duration was <2 months or 2-4 months, growth rate was higher in Ala12Ala than Pro12Pro carriers (differences 9.80 g/week [3.97-15.63] and 6.32 g/week [-1.04 to 13.68], respectively). CONCLUSIONS: The PPARgamma2 Ala12 allele is associated with an increased growth rate in early life. This effect may be influenced by breast-feeding duration. Further studies should replicate these findings, identify the underlying mechanisms, and assess whether these effects persist into later life. PMID- 19188433 TI - Do non-HLA genes influence development of persistent islet autoimmunity and type 1 diabetes in children with high-risk HLA-DR,DQ genotypes? AB - OBJECTIVE: Specific alleles of non-HLA genes INS, CTLA-4, and PTPN22 have been associated with type 1 diabetes. We examined whether some of these alleles influence development of islet autoimmunity or progression from persistent islet autoimmunity to type 1 diabetes in children with high-risk HLA-DR,DQ genotypes. RESEARCH DESIGN AND METHODS: Since 1993, the Diabetes Autoimmunity Study in the Young (DAISY) has followed 2,449 young children carrying HLA-DR,DQ genotypes associated with type 1 diabetes. Of those, 112 have developed islet autoimmunity (persistent autoantibodies to insulin, GAD65, and/or IA-2), and 47 of these have progressed to type 1 diabetes. The influence of polymorphisms of INS(-23Hph1), CTLA-4(T17A), and PTPN22(R620W) on development of persistent islet autoimmunity and progression to type 1 diabetes was evaluated by parametric models and by survival analyses. RESULTS: PTPN22(R620W) allele T was associated with development of persistent islet autoimmunity (hazard ratio 1.83 [95% CI 1.27 2.63]) controlling for ethnicity, presence of HLA-DR3/4,DQB1*0302, and having a first-degree relative with type 1 diabetes. Survival analyses showed a significantly (P = 0.002) higher risk of persistent islet autoimmunity by age 10 years for the TT genotype (27.3%) than for the CT or CC genotype (7.9 and 5.3%, respectively). Cumulative risk of persistent islet autoimmunity was slightly higher (P = 0.02) for the INS(-23Hph1) AA genotype (7.8%) than for the AT or TT genotype (4.2 and 6.4% risk by age 10 years, respectively). CONCLUSIONS: Whereas the HLA-DR3/4,DQB1*0302 genotype had a dramatic influence on both development of islet autoimmunity and progression to type 1 diabetes, the PTPN22(R620W) T allele significantly influences progression to persistent islet autoimmunity in the DAISY cohort. PMID- 19188434 TI - High glucose suppresses epidermal growth factor receptor/phosphatidylinositol 3 kinase/Akt signaling pathway and attenuates corneal epithelial wound healing. AB - OBJECTIVE: Patients with diabetes are at an increased risk for developing corneal complications and delayed wound healing. This study investigated the effects of high glucose on epidermal growth factor receptor (EGFR) signaling and on epithelial wound healing in the cornea. RESEARCH DESIGN AND METHODS: Effects of high glucose on wound healing and on EGFR signaling were investigated in cultured porcine corneas, human corneal epithelial cells, and human corneas using Western blotting and immunofluorescence. Effects of high glucose on reactive oxygen species (ROS) and glutathione levels and on EGFR pathways were assessed in porcine and primary human corneal epithelial cells, respectively. The effects of EGFR ligands and antioxidants on high glucose-delayed epithelial wound healing were assessed in cultured porcine corneas. RESULTS: High glucose impaired ex vivo epithelial wound healing and disturbed cell responses and EGFR signaling to wounding. High glucose suppressed Akt phosphorylation in an ROS-sensitive manner and decreased intracellular glutathione in cultured porcine corneas. Exposure to high glucose for 24 h resulted in an increase in ROS-positive cells in primary human corneal epithelial cells. Whereas heparin-binding EGF-like growth factor and antioxidant N-acetylcysteine had beneficial effects on epithelial wound closure, their combination significantly accelerated high glucose-delayed wound healing to a level similar to that seen in control subjects. Finally, Akt signaling pathway was perturbed in the epithelia of human diabetic corneas, but not in the corneas of nondiabetic, age-matched donors. CONCLUSIONS: High glucose, likely through ROS, impairs the EGFR-phosphatidylinositol 3-kinase/Akt pathway, resulting in delayed corneal epithelial wound healing. Antioxidants in combination with EGFR ligands may be promising potential therapeutics for diabetic keratopathy. PMID- 19188435 TI - Functional targets of the monogenic diabetes transcription factors HNF-1alpha and HNF-4alpha are highly conserved between mice and humans. AB - OBJECTIVE: The evolutionary conservation of transcriptional mechanisms has been widely exploited to understand human biology and disease. Recent findings, however, unexpectedly showed that the transcriptional regulators hepatocyte nuclear factor (HNF)-1alpha and -4alpha rarely bind to the same genes in mice and humans, leading to the proposal that tissue-specific transcriptional regulation has undergone extensive divergence in the two species. Such observations have major implications for the use of mouse models to understand HNF-1alpha- and HNF 4alpha-deficient diabetes. However, the significance of studies that assess binding without considering regulatory function is poorly understood. RESEARCH DESIGN AND METHODS: We compared previously reported mouse and human HNF-1alpha and HNF-4alpha binding studies with independent binding experiments. We also integrated binding studies with mouse and human loss-of-function gene expression datasets. RESULTS: First, we confirmed the existence of species-specific HNF 1alpha and -4alpha binding, yet observed incomplete detection of binding in the different datasets, causing an underestimation of binding conservation. Second, only a minor fraction of HNF-1alpha- and HNF-4alpha-bound genes were downregulated in the absence of these regulators. This subset of functional targets did not show evidence for evolutionary divergence of binding or binding sequence motifs. Finally, we observed differences between conserved and species specific binding properties. For example, conserved binding was more frequently located near transcriptional start sites and was more likely to involve multiple binding events in the same gene. CONCLUSIONS: Despite evolutionary changes in binding, essential direct transcriptional functions of HNF-1alpha and -4alpha are largely conserved between mice and humans. PMID- 19188436 TI - Kinetics of GLUT4 trafficking in rat and human skeletal muscle. AB - OBJECTIVE: In skeletal muscle, insulin stimulates glucose transport activity three- to fourfold, and a large part of this stimulation is associated with a net translocation of GLUT4 from an intracellular compartment to the cell surface. We examined the extent to which insulin or the AMP-activated protein kinase activator AICAR can lead to a stimulation of the exocytosis limb of the GLUT4 translocation pathway and thereby account for the net increase in glucose transport activity. RESEARCH DESIGN AND METHODS: Using a biotinylated photoaffinity label, we tagged endogenous GLUT4 and studied the kinetics of exocytosis of the tagged protein in rat and human skeletal muscle in response to insulin or AICAR. Isolated epitrochlearis muscles were obtained from male Wistar rats. Vastus lateralis skeletal muscle strips were prepared from open muscle biopsies obtained from six healthy men (age 39 +/- 11 years and BMI 25.8 +/- 0.8 kg/m2). RESULTS: In rat epitrochlearis muscle, insulin exposure leads to a sixfold stimulation of the GLUT4 exocytosis rate (with basal and insulin stimulated rate constants of 0.010 and 0.067 min(-1), respectively). In human vastus lateralis muscle, insulin stimulates GLUT4 translocation by a similar sixfold increase in the exocytosis rate constant (with basal and insulin stimulated rate constants of 0.011 and 0.075 min(-1), respectively). In contrast, AICAR treatment does not markedly increase exocytosis in either rat or human muscle. CONCLUSIONS: Insulin stimulation of the GLUT4 exocytosis rate constant is sufficient to account for most of the observed increase in glucose transport activity in rat and human muscle. PMID- 19188437 TI - Ape1/Ref-1 induces glial cell-derived neurotropic factor (GDNF) responsiveness by upregulating GDNF receptor alpha1 expression. AB - Apurinic/apyrimidinic endonuclease 1 (Ape1/Ref-1) dysregulation has been identified in several human tumors and in patients with a variety of neurodegenerative diseases. However, the function of Ape1/Ref-1 is unclear. We show here that Ape1/Ref-1 increases the expression of glial cell-derived neurotropic factor (GDNF) receptor alpha1 (GFRalpha1), a key receptor for GDNF. Expression of Ape1/Ref-1 led to an increase in the GDNF responsiveness in human fibroblast. Ape1/Ref-1 induced GFRalpha1 transcription through enhanced binding of NF-kappaB complexes to the GFRalpha1 promoter. GFRalpha1 levels correlate proportionally with Ape1/Ref-1 in cancer cells. The knockdown of endogenous Ape1/Ref-1 in pancreatic cancer cells markedly suppressed GFRalpha1 expression and invasion in response to GNDF, while overexpression of GFRalpha1 restored invasion. In neuronal cells, the Ape1/Ref-1-mediated increase in GDNF responsiveness not only stimulated neurite outgrowth but also protected the cells from beta-amyloid peptide and oxidative stress. Our results show that Ape1/Ref-1 is a novel physiological regulator of GDNF responsiveness, and they also suggest that Ape1/Ref-1-induced GFRalpha1 expression may play important roles in pancreatic cancer progression and neuronal cell survival. PMID- 19188438 TI - Myocilin is a modulator of Wnt signaling. AB - It is well documented that mutations in the MYOCILIN gene may lead to juvenile- and adult-onset primary open-angle glaucoma. However, the functions of wild-type myocilin are still not well understood. To study the functions of human myocilin and its two proteolytic fragments, these proteins were expressed in HEK293 cells. Conditioned medium from myocilin-expressing cells, as well as purified myocilin, induced the formation of stress fibers in primary cultures of human trabecular meshwork or NIH 3T3 cells. Stress fiber-inducing activity of myocilin was blocked by antibodies against myocilin, as well as secreted inhibitors of Wnt signaling, secreted Frizzled-related protein 1 (sFRP1) or sFRP3, and beta-catenin small interfering RNA. Interaction of myocilin with sFRP1, sFRP3, and several Frizzled receptors was confirmed by immunoprecipitation experiments and by binding of myocilin to the surface of cells expressing cysteine-rich domains of different Frizzled and sFRPs. Treatment of NIH 3T3 cells with myocilin and its fragments induced intracellular redistribution of beta-catenin and its accumulation on the cellular membrane but did not induce nuclear accumulation of beta-catenin. Overexpression of myocilin in the eye angle tissues of transgenic mice stimulated accumulation of beta-catenin in these tissues. Myocilin and Wnt proteins may perform redundant functions in the mammalian eye, since myocilin modulates Wnt signaling by interacting with components of this signaling pathway. PMID- 19188439 TI - MicroRNA-1 negatively regulates expression of the hypertrophy-associated calmodulin and Mef2a genes. AB - Calcium signaling is a central regulator of cardiomyocyte growth and function. Calmodulin is a critical mediator of calcium signals. Because the amount of calmodulin within cardiomyocytes is limiting, the precise control of calmodulin expression is important for the regulation of calcium signaling. In this study, we show for the first time that calmodulin levels are regulated posttranscriptionally in heart failure. The cardiomyocyte-restricted microRNA miR 1 inhibited the translation of calmodulin-encoding mRNAs via highly conserved target sites within their 3' untranslated regions. In keeping with its effect on calmodulin expression, miR-1 downregulated calcium-calmodulin signaling through calcineurin to NFAT. miR-1 also negatively regulated the expression of Mef2a and Gata4, key transcription factors that mediate calcium-dependent changes in gene expression. Consistent with the downregulation of these hypertrophy-associated genes, miR-1 attenuated cardiomyocyte hypertrophy in cultured neonatal rat cardiomyocytes and in the intact adult heart. Our data indicate that miR-1 regulates cardiomyocyte growth responses by negatively regulating the calcium signaling components calmodulin, Mef2a, and Gata4. PMID- 19188440 TI - Association of C-terminal ubiquitin hydrolase BRCA1-associated protein 1 with cell cycle regulator host cell factor 1. AB - Protein ubiquitination provides an efficient and reversible mechanism to regulate cell cycle progression and checkpoint control. Numerous regulatory proteins direct the addition of ubiquitin to lysine residues on target proteins, and these are countered by an army of deubiquitinating enzymes (DUBs). BRCA1-associated protein-1 (Bap1) is a ubiquitin carboxy-terminal hydrolase and is frequently mutated in lung and sporadic breast tumors. Bap1 can suppress growth of lung cancer cells in athymic nude mice and this requires its DUB activity. We show here that Bap1 interacts with host cell factor 1 (HCF-1), a transcriptional cofactor found in a number of important regulatory complexes. Bap1 binds to the HCF-1 beta-propeller using a variant of the HCF-binding motif found in herpes simplex virus VP16 and other HCF-interacting proteins. HCF-1 is K48 and K63 ubiquitinated, with a major site of linkage at lysines 1807 and 1808 in the HCF 1(C) subunit. Expression of a catalytically inactive version of Bap1 results in the selective accumulation of K48 ubiquitinated polypeptides. Depletion of Bap1 using small interfering RNA results in a modest accumulation of HCF-1(C), suggesting that Bap1 helps to control cell proliferation by regulating HCF-1 protein levels and by associating with genes involved in the G(1)-S transition. PMID- 19188441 TI - Distinct protein arginine methyltransferases promote ATP-dependent chromatin remodeling function at different stages of skeletal muscle differentiation. AB - Temporal regulation of gene expression is a hallmark of cellular differentiation pathways, yet the mechanisms controlling the timing of expression for different classes of differentiation-specific genes are not well understood. We previously demonstrated that the class II arginine methyltransferase Prmt5 was required for skeletal muscle differentiation at the early stages of myogenesis (C. S. Dacwag, Y. Ohkawa, S. Pal, S. Sif, and A. N. Imbalzano, Mol. Cell. Biol. 27:384-394, 2007). Specifically, when Prmt5 levels were reduced, the ATP-dependent SWI/SNF chromatin-remodeling enzymes could not interact with or remodel the promoter of myogenin, an essential early gene. Here we investigated the requirement for Prmt5 and the class I arginine methyltransferase Carm1/Prmt4 in the temporal control of myogenesis. Both arginine methyltransferases could bind to and modify histones at late-gene regulatory sequences. However, the two enzymes showed sequential requirements for gene expression. Prmt5 was required for early-gene expression but dispensable for late-gene expression. Carm1/Prmt4 was required for late- but not for early-gene expression. The reason for the requirement for Carm1/Prmt4 at late genes was to facilitate SWI/SNF chromatin-remodeling enzyme interaction and remodeling at late-gene loci. Thus, distinct arginine methyltransferases are employed at different times of skeletal muscle differentiation for the purpose of facilitating ATP-dependent chromatin-remodeling enzyme interaction and function at myogenic genes. PMID- 19188442 TI - KSR1 modulates the sensitivity of mitogen-activated protein kinase pathway activation in T cells without altering fundamental system outputs. AB - Mitogen-activated protein kinase (MAPK) cascades are evolutionarily conserved signaling pathways that regulate cell fate decisions. They generate a wide range of signal outputs, including graded and digital responses. In T cells, MAPK activation is digital in response to T-cell-receptor stimulation; however, whether other receptors on T cells that lead to MAPK activation are graded or digital is unknown. Here we evaluate MAPK activation in T cells at the single cell level. We show that T cells responded digitally to stimulation with superantigen-loaded antigen-presenting cells, whereas they responded in a graded manner to the chemokine SDF-1, demonstrating that the system output of the MAPK module is highly plastic and determined by components upstream of the MAPK module. These findings also confirm that different MAPK system outputs are used by T cells to control discrete biological functions. Scaffold proteins are essential for proper MAPK signaling and function as they physically assemble multiple components and regulators of MAPK cascades. We found that the scaffold protein KSR1 regulated the threshold required for MAPK activation in T cells without affecting the nature of the response. We conclude that KSR1 plays a central role in determining the sensitivity of T-cell responses and is thus well positioned as a key control point. PMID- 19188443 TI - Evidence for regulation of mitotic progression through temporal phosphorylation and dephosphorylation of CK2alpha. AB - Proper mitotic progression is crucial for maintenance of genomic integrity in proliferating cells and is regulated through an intricate series of events, including protein phosphorylation governed by a complex network of protein kinases. One kinase family implicated in the regulation of mitotic progression is protein kinase CK2, a small family of enzymes that is overexpressed in cancer and induces transformation in mice and cultured fibroblasts. CK2alpha, one isoform of the catalytic subunits of CK2, is maximally phosphorylated at four sites in nocodazole-treated cells. To investigate the effects of CK2alpha phosphorylation on mitotic progression, we generated phosphospecific antibodies against its mitotic phosphorylation sites. In U2OS cells released from S-phase arrest, these antibodies reveal that CK2alpha is most highly phosphorylated in prophase and metaphase. Phosphorylation gradually decreases during anaphase and becomes undetectable during telophase and cytokinesis. Stable expression of phosphomimetic CK2alpha (CK2alpha-4D, CK2alpha-4E) results in aberrant centrosome amplification and chromosomal segregation defects and loss of mitotic cells through mitotic catastrophe. Conversely, cells expressing nonphosphorylatable CK2alpha (CK2alpha-4A) show a decreased ability to arrest in mitosis following nocodazole treatment, suggesting involvement in the spindle assembly checkpoint. Collectively, these studies indicate that reversible phosphorylation of CK2alpha requires precise regulation to allow proper mitotic progression. PMID- 19188444 TI - Two classes of dosage compensation complex binding elements along Caenorhabditis elegans X chromosomes. AB - Dosage compensation equalizes X-linked gene products between the sexes. In Caenorhabditis elegans, the dosage compensation complex (DCC) binds both X chromosomes in XX animals and halves the transcription from each. The DCC is recruited to the X chromosomes by a number of loci, rex sites, and is thought to spread from these sites by an unknown mechanism to cover the rest of the chromosome. Here we describe a novel class of DCC-binding elements that we propose serve as "way stations" for DCC binding and spreading. Both rex sites and way stations comprise strong foci of DCC binding on the native X chromosome. However, rex sites maintain their ability to bind large amounts of DCC even on X duplications detached from the native X, while way stations do not. These results suggest that two distinct classes of DCC-binding elements facilitate recruitment and spreading of the DCC along the X chromosome. PMID- 19188445 TI - APE1/Ref-1 interacts with NPM1 within nucleoli and plays a role in the rRNA quality control process. AB - APE1/Ref-1 (hereafter, APE1), a DNA repair enzyme and a transcriptional coactivator, is a vital protein in mammals. Its role in controlling cell growth and the molecular mechanisms that fine-tune its different cellular functions are still not known. By an unbiased proteomic approach, we have identified and characterized several novel APE1 partners which, unexpectedly, include a number of proteins involved in ribosome biogenesis and RNA processing. In particular, a novel interaction between nucleophosmin (NPM1) and APE1 was characterized. We observed that the 33 N-terminal residues of APE1 are required for stable interaction with the NPM1 oligomerization domain. As a consequence of the interaction with NPM1 and RNA, APE1 is localized within the nucleolus and this localization depends on cell cycle and active rRNA transcription. NPM1 stimulates APE1 endonuclease activity on abasic double-stranded DNA (dsDNA) but decreases APE1 endonuclease activity on abasic single-stranded RNA (ssRNA) by masking the N terminal region of APE1 required for stable RNA binding. In APE1-knocked-down cells, pre-rRNA synthesis and rRNA processing were not affected but inability to remove 8-hydroxyguanine-containing rRNA upon oxidative stress, impaired translation, lower intracellular protein content, and decreased cell growth rate were found. Our data demonstrate that APE1 affects cell growth by directly acting on RNA quality control mechanisms, thus affecting gene expression through posttranscriptional mechanisms. PMID- 19188447 TI - p31 deficiency influences endoplasmic reticulum tubular morphology and cell survival. AB - p31, the mammalian orthologue of yeast Use1p, is an endoplasmic reticulum (ER) localized soluble N-ethylmaleimide-sensitive factor attachment protein (SNAP) receptor (SNARE) that forms a complex with other SNAREs, particularly syntaxin 18. However, the role of p31 in ER function remains unknown. To determine the role of p31 in vivo, we generated p31 conditional knockout mice. We found that homozygous deletion of the p31 gene led to early embryonic lethality before embryonic day 8.5. Conditional knockout of p31 in brains and mouse embryonic fibroblasts (MEFs) caused massive apoptosis accompanied by upregulation of ER stress-associated genes. Microscopic analysis showed vesiculation and subsequent enlargement of the ER membrane in p31-deficient cells. This type of drastic disorganization in the ER tubules has not been demonstrated to date. This marked change in ER structure preceded nuclear translocation of the ER stress-related transcription factor C/EBP homologous protein (CHOP), suggesting that ER stress induced apoptosis resulted from disruption of the ER membrane structure. Taken together, these results suggest that p31 is an essential molecule involved in the maintenance of ER morphology and that its deficiency leads to ER stress-induced apoptosis. PMID- 19188446 TI - A tautomerase-null macrophage migration-inhibitory factor (MIF) gene knock-in mouse model reveals that protein interactions and not enzymatic activity mediate MIF-dependent growth regulation. AB - Macrophage migration-inhibitory factor (MIF) is an upstream regulator of innate immunity and a potential molecular link between inflammation and cancer. The unusual structural homology between MIF and certain tautomerases, which includes both a conserved substrate-binding pocket and a catalytic N-terminal proline (Pro1), has fueled speculation that an enzymatic reaction underlies MIF's biologic function. To address the functional role of the MIF tautomerase activity in vivo, we created a knock-in mouse in which the endogenous mif gene was replaced by one encoding a tautomerase-null, Pro1-->Gly1 MIF protein (P1G-MIF). While P1G-MIF is completely inactive catalytically, it maintains significant, albeit reduced, binding to its cell surface receptor (CD74) and to the intracellular binding protein JAB1/CSN5. P1G-MIF knock-in mice (mif(P1G/P1G)) and cells derived from these mice show a phenotype in assays of growth control and tumor induction that is intermediate between those of the wild type (mif(+/+)) and complete MIF deficiency (mif(-)(/)(-)). These data provide genetic evidence that MIF's intrinsic tautomerase activity is dispensable for this cytokine's growth-regulatory properties and support a role for the N-terminal region in protein-protein interactions. PMID- 19188448 TI - The essential N terminus of the Pta1 scaffold protein is required for snoRNA transcription termination and Ssu72 function but is dispensable for pre-mRNA 3' end processing. AB - Saccharomyces cerevisiae Pta1 is a component of the cleavage/polyadenylation factor (CPF) 3'-end processing complex and functions in pre-mRNA cleavage, poly(A) addition, and transcription termination. In this study, we investigated the role of the N-terminal region of Pta1 in transcription and processing. We report that a deletion of the first 75 amino acids (pta1-Delta75) causes thermosensitive growth, while the deletion of an additional 25 amino acids is lethal. The pta1-Delta75 mutant is defective for snoRNA termination, RNA polymerase II C-terminal domain Ser5-P dephosphorylation, and gene looping but is fully functional for mRNA 3'-end processing. Furthermore, different regions of Pta1 interact with the CPF subunits Ssu72, Pti1, and Ysh1, supporting the idea that Pta1 acts as a scaffold to organize CPF. The first 300 amino acids of Pta1 are sufficient for interactions with Ssu72, which is needed for pre-mRNA cleavage. By the degron-mediated depletion of Pta1, we show that the removal of this essential region leads to a loss of Ssu72, yet surprisingly, in vitro cleavage and polyadenylation remain efficient. In addition, a fragment containing amino acids 1 to 300 suppresses 3'-end processing in wild-type extracts. These findings suggest that the amino terminus of Pta1 has an inhibitory effect and that this effect can be neutralized through the interaction with Ssu72. PMID- 19188449 TI - hSirT1-dependent regulation of the PCAF-E2F1-p73 apoptotic pathway in response to DNA damage. AB - The NAD(+)-dependent histone deacetylase hSirT1 regulates cell survival and stress responses by inhibiting p53-, NF-kappaB-, and E2F1-dependent transcription. Here we show that the hSirT1/PCAF interaction controls the E2F1/p73 apoptotic pathway. hSirT1 represses E2F1-dependent P1p73 promoter activity in untreated cells and inhibits its activation in response to DNA damage. hSirT1, PCAF, and E2F1 are corecruited in vivo on theP1p73 promoter. hSirT1 deacetylates PCAF in vitro and modulates PCAF acetylation in vivo. In cells exposed to apoptotic DNA damage, nuclear NAD(+) levels decrease and inactivate hSirT1 without altering the hSirT1 interaction with PCAF and hSirT1 binding to the P1p73 promoter. The reactivation of hSirT1 by pyruvate that increases the [NAD(+)]/[NADH] ratio completely abolished the DNA damage-induced activation of TAp73 expression, thus linking the modulation of chromatin-bound hSirT1 deacetylase activity by the intracellular redox state with P1p73 promoter activity. The release of PCAF from hSirT1 repression favors the assembly of transcriptionally active PCAF/E2F1 complexes onto the P1p73 promoter and p53 independent apoptosis. Our results identify hSirT1 and PCAF as potential targets to modulate tumor cell survival and chemoresistance irrespective of p53 status. PMID- 19188450 TI - Dax-1 and steroid receptor RNA activator (SRA) function as transcriptional coactivators for steroidogenic factor 1 in steroidogenesis. AB - The nuclear receptor steroidogenic factor 1 (SF-1) is essential for adrenal development and steroidogenesis. The atypical orphan nuclear receptor Dax-1 binds to SF-1 and represses SF-1 target genes. Paradoxically, however, loss-of-function mutations of Dax-1 also cause adrenal hypoplasia, suggesting that Dax-1 may function as an SF-1 coactivator under some circumstances. Indeed, we found that Dax-1 can function as a dosage-dependent SF-1 coactivator. Both SF-1 and Dax-1 bind to steroid receptor RNA activator (SRA), a coactivator that functions as an RNA. The coactivator TIF2 also associates with Dax-1 and synergistically coactivates SF-1 target gene transcription. A naturally occurring Dax-1 mutation inhibits this transactivation, and the mutant Dax-1-TIF2 complex mislocalizes in living cells. Coactivation by Dax-1 is abolished by SRA knockdown. The expression of the steroidogenic gene products steroidogenic acute regulatory protein (StAR) and melanocortin 2 receptor is reduced in adrenal Y1 cells following the knockdown of endogenous SRA. Similarly, the knockdown of endogenous Dax-1 downregulates the expression of the steroidogenic gene products CYP11A1 and StAR in both H295R adrenal and MA-10 Leydig cells. These findings reveal novel functions of SRA and Dax-1 in steroidogenesis and adrenal biology. PMID- 19188451 TI - Estrogen receptor alpha represses transcription of early target genes via p300 and CtBP1. AB - The regulation of gene expression by nuclear receptors controls the phenotypic properties and diverse biologies of target cells. In breast cancer cells, estrogen receptor alpha (ERalpha) is a master regulator of transcriptional stimulation and repression, yet the mechanisms by which agonist-bound ERalpha elicits repression are poorly understood. We analyzed early estrogen-repressed genes and found that ERalpha is recruited to ERalpha binding sites of these genes, albeit more transiently and less efficiently than for estrogen-stimulated genes. Of multiple cofactors studied, only p300 was recruited to ERalpha binding sites of repressed genes, and its knockdown prevented estrogen-mediated gene repression. Because p300 is involved in transcription initiation, we tested whether ERalpha might be trying to stimulate transcription at repressed genes, with ultimately failure and a shift to a repressive program. We found that estrogen increases transcription in a rapid but transient manner at early estrogen-repressed genes but that this is followed by recruitment of the corepressor CtBP1, a p300-interacting partner that plays an essential role in the repressive process. Thus, at early estrogen-repressed genes, ERalpha initiates transient stimulation of transcription but fails to maintain the transcriptional process observed at estrogen-stimulated genes; rather, it uses p300 to recruit CtBP1-containing complexes, eliciting chromatin modifications that lead to transcriptional repression. PMID- 19188455 TI - MARQulS: quality improvement strategies for European cross-border healthcare. PMID- 19188452 TI - Stimulation of polo-like kinase 3 mRNA decay by tristetraprolin. AB - Polo-like protein kinase 3 (Plk3) has been proposed to regulate entry into S phase and promote apoptosis in response to oxidative stress. Its mRNA contains three AU-rich elements (AREs) in its 3' untranslated region (3'-UTR) that can contribute to the rapid degradation of labile transcripts. We investigated the possibility that tristetraprolin (TTP), a tandem CCCH zinc finger protein, could promote the decay of Plk3 transcripts. TTP is known to stimulate the deadenylation and decay of mRNAs possessing one or more copies of the consensus nonamer motif UUAUUUAUU. In stable mouse fibroblast cell lines derived from wild type and TTP knockout littermates, the decay of Plk3 transcripts after serum stimulation was slowed in the absence of TTP. The specificity of TTP for promoting the degradation of Plk3 was demonstrated by the unaltered decay of Plk3 mRNA in cell lines deficient in the TTP family members ZFP36L1 and ZFP36L2. We also found that the AREs present in the Plk3 transcript were essential for both the binding of TTP to the 3'-UTR and promoting the destruction of target transcripts in cotransfection experiments. The regulation of Plk3 mRNA stability by TTP may influence the control of the cell cycle by this protein kinase. PMID- 19188456 TI - Quality requirements for cross-border care in Europe: a qualitative study of patients', professionals' and healthcare financiers' views. AB - BACKGROUND: In the past decade the issue of patient mobility has emerged on the European health policy agenda. Although the volume of patients crossing borders to obtain healthcare is low, it is increasing continuously and, due to its legal, financial and medical implications, has generated considerable interest among health policy and other decision makers. However, there is little information available on the safety and patient-centredness of cross-border care and neither governments nor citizens have an explicit basis for comparing healthcare delivery in Europe. METHODS: This study investigated the viewpoints of patients, professionals and healthcare financiers on the safety and patient-centredness of cross-border care. Qualitative interviews were carried out during 2005 and early 2006 with 40 patients, 30 professionals (doctors, nurses and managers) and 3 healthcare-financing bodies. RESULTS: Although cross-border care has become a common issue in many European countries, there remain uncertainties on the side of each of the parties addressed--patients, professionals and financiers--with regard to the provision of cross-border care. One of the most striking results of this project is the current lack of research on systematic knowledge on the quality of cross-border care. CONCLUSION: Many of the issues identified through this research may have a potential impact on the quality and safety of cross border care and will support further investigation and help shape the health policy agenda on patients crossing borders in European Union countries. PMID- 19188457 TI - National quality improvement policies and strategies in European healthcare systems. AB - OBJECTIVE: This survey provides an overview of the development of policies and strategies for quality improvement in European healthcare systems, by mapping quality improvement policies and strategies, progress in their implementation, and early indications of their impact. STUDY DESIGN: A survey of quality improvement policies and strategies in healthcare systems of the European Union was conducted in 2005 for the first phase of the Methods of Assessing Response to Quality Improvement Strategies (MARQuIS) project. PARTICIPANTS: The survey, completed by 68 key experts in quality improvement from 24 European Union member states, represents their views and accounts of quality improvement policies and strategies in their healthcare systems. PRINCIPAL FINDINGS: There are substantial international and intra-national variations in the development of healthcare quality improvement. Legal requirements for quality improvement strategies are an important driver of progress, along with the activities of national governments and professional associations and societies. Patient and service user organisations appear to have less influence on quality improvement. Wide variation in voluntary and mandatory coverage of quality improvement policies and strategies across sectors can potentially lead to varying levels of progress in implementation. Many healthcare organisations lack basic infrastructure for quality improvement. CONCLUSIONS: Some convergence can be observed in policies on quality improvement in healthcare. Nevertheless, the growth of patient mobility across borders, along with the implications of free market provisions for the organisation and funding of healthcare systems in European Union member states, require policies for cooperation and learning transfer. PMID- 19188459 TI - Cross-border care and healthcare quality improvement in Europe: the MARQuIS research project. AB - Citizens are increasingly crossing borders within the European Union (EU). Europeans have always been free to travel to receive care abroad, but if they wished to benefit from their statutory social protection scheme, they were subject to their local or national legislation on social protection. This changed in 1991 with the European Court of Justice defining healthcare as a service, starting a debate on the right balance between different principles in European treaties: movement of persons, goods and services, versus the responsibility of member states to organise their healthcare systems. Simultaneously, cross-border cooperation has developed between member states. In this context, patient mobility has become a relevant issue on the EU's agenda. The EU funded a number of Scientific Support to Policies (SSP) activities within the Sixth Framework Programme, to provide the evidence needed by EU policy makers to deal with issues that European citizens face due to enhanced mobility in Europe. One SSP project "Methods of Assessing Response to Quality Improvement Strategies" (MARQuIS), focused on cross-border care. It aimed to assess the value of different quality strategies, and to provide information needed when: (1) countries contract care for patients moving across borders; and (2) individual hospitals review the design of their quality strategies. This article describes the European context related to healthcare, and its implications for cross-border healthcare in Europe. The background information demonstrates a need for further research and development in this area. PMID- 19188458 TI - Application of quality improvement strategies in 389 European hospitals: results of the MARQuIS project. AB - CONTEXT: This study was part of the Methods of Assessing Response to Quality Improvement Strategies (MARQuIS) research project investigating the impact of quality improvement strategies on hospital care in various countries of the European Union (EU), in relation to specific needs of cross-border patients. AIM: This paper describes how EU hospitals have applied seven quality improvement strategies previously defined by the MARQuIS study: organisational quality management programmes; systems for obtaining patients' views; patient safety systems; audit and internal assessment of clinical standards; clinical and practice guidelines; performance indicators; and external assessment. METHODS: A web-based questionnaire was used to survey acute care hospitals in eight EU countries. The reported findings were later validated via on-site survey and site visits in a sample of the participating hospitals. Data collection took place from April to August 2006. RESULTS: 389 hospitals participated in the survey; response rates varied per country. All seven quality improvement strategies were widely used in European countries. Activities related to external assessment were the most broadly applied across Europe, and activities related to patient involvement were the least widely implemented. No one country implemented all quality strategies at all hospitals. There were no differences between participating hospitals in western and eastern European countries regarding the application of quality improvement strategies. CONCLUSIONS: Implementation varied per country and per quality improvement strategy, leaving considerable scope for progress in quality improvements. The results may contribute to benchmarking activities in European countries, and point to further areas of research to explore the relationship between the application of quality improvement strategies and actual hospital performance. PMID- 19188460 TI - Differentiating between hospitals according to the "maturity" of quality improvement systems: a new classification scheme in a sample of European hospitals. AB - AIM: This study, part of the Methods of Assessing Response to Quality Improvement Strategies (MARQuIS) research project focusing on cross-border patients in Europe, investigated quality policies and improvement in healthcare systems across the European Union (EU). The aim was to develop a classification scheme for the level of quality improvement (maturity) in EU hospitals, in order to evaluate hospitals according to the maturity of their quality improvement activities. METHODS: A web-based questionnaire survey designed to measure quality improvement in EU hospitals was used as the basis for the classification scheme. Items included for the development of an evaluation tool--the maturity index- were considered important contributors to quality improvement. The four-stage quality cycle (plan, do, check and act) was used to determine the level of maturity of the various items. Psychometric properties of the classification scheme were assessed, and validation analyses were performed. RESULTS: A total of 389 hospitals participated in a questionnaire survey; response rates varied by country. For a final sample of 349 hospitals, it was possible to construct a quality improvement maturity index which consisted of seven domains and 113 items. The results of independent analyses sustained the validity of the index, which was useful in differentiating between hospitals in the research sample according to the maturity of their quality improvement system (defined as the total of all quality improvement activities). DISCUSSION: Further research is recommended to develop an instrument which for use in the future as a practical tool to evaluate the maturity of hospital quality improvement systems. PMID- 19188461 TI - Is patient-centredness in European hospitals related to existing quality improvement strategies? Analysis of a cross-sectional survey (MARQuIS study). AB - BACKGROUND: There is growing recognition of patients' contributions to setting objectives for their own care, improving health outcomes and evaluating care. OBJECTIVE: To quantify the extent to which European hospitals have implemented strategies to promote a patient-centred approach, and to assess whether these strategies are associated with hospital characteristics and the development of the hospital's quality improvement system. DESIGN: Cross-sectional survey of 351 European hospital managers and professionals. MAIN OUTCOME MEASURES: Patients' rights, patient information and empowerment, patient involvement in quality management, learning from patients, and patient hotel services at the hospital and ward level were assessed. The hypothesis that the implementation of strategies to improve patient-centredness is associated with hospital characteristics, including maturity of the hospital's quality management system, was tested using binary logistic regression. RESULTS: In general, hospitals reported high implementation of policies for patients' rights (85.5%) and informed consent (93%), whereas strategies to involve patients (71%) and learn from their experience (66%) were less frequently implemented. For 13 out of 18 hospital strategies, institutions with a more developed quality improvement system consistently reported better results (percentage differences within maturity classification ranged from 12.4% to 46.6%). The strength of association between implementation of patient-centredness strategies and the quality improvement system, however, seemed lower at the ward than at the hospital level. Some associations (OR 2.1 to 5.1) disappeared or were weaker after adjustment for potential confounding variables (OR 2.2 to 3.7). CONCLUSIONS: Although quality improvement systems seem to be effective with regard to the implementation of selected patient-centredness strategies, they seem to be insufficient to ensure widespread implementation of patient-centredness throughout the organisation. PMID- 19188462 TI - Do European hospitals have quality and safety governance systems and structures in place? AB - Internal systems for quality and safety were assessed in 89 hospitals in six European states, by external teams using standardised criteria and procedures, as part of the Methods of Assessing Response to Quality Improvement Strategies (MARQuIS) project. The assessments were made primarily to identify the current use of quality management systems in the sample hospitals, and also to demonstrate a potential tool for comparable assessment of hospitals in general. The large majority of the hospitals had a formal, documented infrastructure to manage quality and safety, but a significant minority had no designated mission, programme or coordination. In two-thirds of hospitals, the governing body was active in defining policy and programmes for improvement, and received reports on quality, safety and patient satisfaction at least once a year. The brief on-site assessments identified systematic variations, within and between countries, in structures and processes of governance and to document the uptake of best practice. Unacceptable variations in practice could be reduced, to the benefit of consumers and providers, by developing and publishing basic organisational standards relevant to all European states. The simple assessment criteria designed for this project could be developed into a practical tool for self assessment, peer review or benchmarking of hospitals across national borders. This assessment, combined with explicit, relevant and achievable standards, could provide a vehicle to promote the voluntary uptake of best practice and consistency in quality and safety among hospitals in Europe. PMID- 19188463 TI - Implementation of patient safety strategies in European hospitals. AB - CONTEXT: This study is part of the Methods of Assessing Response to Quality Improvement Strategies (MARQuIS) research project on cross-border care, investigating quality improvement strategies in healthcare systems across the European Union (EU). AIM: To explore to what extent a sample of acute care European hospitals have implemented patient safety strategies and mechanisms and whether the implementation is related to the type of hospital. METHODS: Data were collected on patient safety structures and mechanisms in 389 acute care hospitals in eight EU countries using a web-based questionnaire. Subsequently, an on-site audit was carried out by independent surveyors in 89 of these hospitals to assess patient safety outputs. This paper presents univariate and bivariate statistics on the implementation and explores the associations between implementation of patient safety strategies and hospital type using the chi(2) test and Fisher exact test. RESULTS: Structures and plans for safety (including responsibilities regarding patient safety management) are well developed in most of the hospitals that participated in this study. The study found greater variation regarding the implementation of mechanisms or activities to promote patient safety, such as electronic drug prescription systems, guidelines for prevention of wrong patient, wrong site and wrong surgical procedure, and adverse events reporting systems. In the sample of hospitals that underwent audit, a considerable proportion do not comply with basic patient safety strategies--for example, using bracelets for adult patient identification and correct labelling of medication. PMID- 19188464 TI - Impact of quality strategies on hospital outputs. AB - CONTEXT: This study was part of the Methods of Assessing Response to Quality Improvement Strategies (MARQuIS) research project on patients crossing borders, a study to investigate quality improvement strategies in healthcare systems across the European Union (EU). AIM: To explore the association between the implementation of quality improvement strategies in hospitals and hospitals' success in meeting defined quality requirements that are considered intermediate outputs of the care process. METHODS: Data regarding the implementation of seven quality improvement strategies (accreditation, organisational quality management programmes, audit and internal assessment of clinical standards, patient safety systems, clinical practice guidelines, performance indicators and systems for obtaining patients' views) and four dimensions of outputs (clinical, safety, patient-centredness and cross-border patient-centredness) were collected from 389 acute care hospitals in eight EU countries using a web-based questionnaire. In a second phase, 89 of these hospitals participated in an on-site audit by independent surveyors. Pearson correlation and linear regression models were used to explore associations and relations between quality improvement strategies and achievement of outputs. RESULTS: Positive associations were found between six internal quality improvement strategies and hospital outputs. The quality improvement strategies could be reasonably subsumed under one latent index which explained about half of their variation. The analysis of outputs concluded that the outputs can also be considered part of a single construct. The findings indicate that the implementation of internal as well as external quality improvement strategies in hospitals has beneficial effects on the hospital outputs studied here. CONCLUSION: The implementation of internal quality improvement strategies as well as external assessment systems should be promoted. PMID- 19188465 TI - Learning from MARQuIS: future direction of quality and safety in hospital care in the European Union. AB - This article summarises the significant lessons to be drawn from, and the policy implications of, the findings of the Methods of Assessing Response to Quality Improvement Strategies (MARQuIS) project--a part of the suite of research projects intended to support policy established by the European Commission through its Sixth Framework Programme. The article first reviews the findings of MARQuIS and their implications for healthcare providers (and particularly for hospitals), and then addresses the broader policy implications for member states of the European Union (EU) and for the commission itself. Against the background of the European Commission's Seventh Framework Programme, it then outlines a number of future areas for research to inform policy and practice in quality and safety in Europe. The article concludes that at this stage, a unique EU-wide quality improvement system for hospitals does not seem to be feasible or effective. Because of possible future community action in this field, attention should focus on the use of existing research on quality and safety strategies in healthcare, with the aim of combining soft measures to accelerate mutual learning. Concrete measures should be considered only in areas for which there is substantial evidence and effective implementation can be ensured. PMID- 19188467 TI - Volume and diagnosis: an approach to cross-border care in eight European countries. AB - OBJECTIVES: Mobility of patients is a pertinent issue on the European Union's agenda. This study aimed to estimate the volume and main diagnoses of cross border care in eight European countries, in order to provide policy makers with background information about the nature of patient mobility in Europe. METHODS: This article reports the combined findings from three independent studies that compiled self-reported information on admissions data and main diagnoses from more than 200 hospitals in eight European countries. RESULTS: The average volume of cross-border patients accounted for less than 1% of total admissions in the hospitals studied here. Diseases of the circulatory system (mainly acute myocardial infarction) and fractures were the most common reasons for hospitalisation of European patients abroad. Deliveries and other diagnoses related to pregnancy, pneumonia, appendicitis and other diseases of the digestive system, aftercare procedures, and disorders of the eye and adnexa were also common diagnoses for this population. CONCLUSIONS: Hospitals should reinforce their efforts to adapt the care provided to the needs of foreign patients in treatment areas that cover the most frequent pathologies identified in this population. PMID- 19188469 TI - Intimal hyperplasia in rats after subcutaneous injection of a somatostatin analog. AB - The somatostatin analog octreotide was administered to male and female Sprague Dawley rats by subcutaneous injection for thirteen weeks at 0 (saline control), 0 (placebo control [mannitol and lactic acid; pH 4.2]), 1.25 mg/kg/day and 2.5 mg/kg/day to explore its potential effect on cutaneous vascular morphology. The placebo caused an increase in the incidence of intimal hyperplasia compared to saline controls in female rats; octreotide increased the incidence and severity of intimal hyperplasia in males and females. Intimal hyperplasia consisted of increased numbers of cells located between the endothelial cell layer and the internal elastic lamina. Severity was based on the degree of compromise of the vascular lumen (regardless of vessel size and number), with severely affected vessels having no visible lumen. Intimal hyperplasia in rats treated with octreotide was considered to be an unexpected and adverse finding, given that this compound and other somatostatin analogs have been investigated as reducers of intimal proliferation or restenosis after angioplasty in humans and that no such lesion has been reported in the literature for this class of compound to date. The induction of intimal hyperplasia by the placebo is also a notable finding; this may be because of the low pH of the formulation. PMID- 19188468 TI - Phytol-induced hepatotoxicity in mice. AB - Phytanic acid is a branched-chain, saturated fatty acid present in high concentrations in dairy products and ruminant fat. Some other dietary fats contain lower levels of phytol, which is readily converted to phytanic acid after absorption. Phytanic acid is a peroxisome proliferator binding the nuclear transcription factor peroxisome proliferator-activated receptor alpha (PPARalpha) to induce expression of genes encoding enzymes of fatty acid oxidation in peroxisomes and mitochondria. Administration of dietary phytol (0.5% or 1%) to normal mice for twelve to eighteen days caused consistent PPARalpha-mediated responses, such as lower body weights, higher liver weights, peroxisome proliferation, increased catalase expression, and hepatocellular hypertrophy and hyperplasia. Female mice fed 0.5% phytol and male and female mice fed 1% phytol exhibited midzonal hepatocellular necrosis, periportal hepatocellular fatty vacuolation, and corresponding increases in liver levels of the phytol metabolites phytanic acid and pristanic acid. Hepatic expression of sterol carrier protein-x (SCP-x) was five- to twelve-fold lower in female mice than in male mice. These results suggest that phytol may cause selective midzonal hepatocellular necrosis in mice, an uncommon pattern of hepatotoxic injury, and that the greater susceptibility of female mice may reflect a lower capacity to oxidize phytanic acid because of their intrinsically lower hepatic expression of SCP-x. PMID- 19188470 TI - The battle of the clot. PMID- 19188471 TI - Intensive therapy of type 2 diabetes (ACCORD trial) (OCTOBER 2008). PMID- 19188472 TI - Intensive therapy of type 2 diabetes (ACCORD trial) (OCTOBER 2008). PMID- 19188473 TI - Long QT syndrome: (AUGUST 2008). PMID- 19188474 TI - A facial cutaneous horn. PMID- 19188475 TI - Autosomal dominant polycystic kidney disease: emerging concepts of pathogenesis and new treatments. AB - Some of the mystery of autosomal dominant polycystic kidney disease (ADPKD) is starting to clear. Basic research is shedding light on its pathogenesis, and new treatments are in clinical trials. This paper reviews some of these advances and what they mean to patients. PMID- 19188476 TI - Role of barium esophagography in evaluating dysphagia. AB - Patients with dysphagia can initially undergo either endoscopic or radiologic evaluation, depending on the clinical history and physician preference. We usually recommend that most patients with dysphagia initially undergo barium esophagography, and in this paper we discuss its role in evaluating dysphagia. PMID- 19188477 TI - Anticoagulants and pregnancy: when are they safe? AB - Prescribing anticoagulants to pregnant women can be difficult and stressful. Fortunately, low-molecular-weight heparins (LMWHs) and unfractionated heparin are quite safe and efficacious when properly selected, dosed, and monitored. Maternal and fetal concerns must be considered at all times, with a careful assessment of the risks and benefits of anticoagulant therapy in each patient. Further research should help to clarify who should receive thromboprophylaxis, how to prevent adverse pregnancy outcomes in women with various thrombophilias, and how best to treat pregnant women who have a prosthetic heart valve. PMID- 19188478 TI - Cancer and clots: all cases of venous thromboembolism are not treated the same. AB - Idiopathic venous thromboembolism (VTE) can be the first sign of cancer, although how extensively one should search for cancer in a patient with idiopathic VTE is not clear. Treating VTE is more complex in cancer patients than in those without cancer. The authors discuss their approach to searching for undiagnosed cancer in patients with idiopathic VTE and to managing VTE in patients with cancer. PMID- 19188480 TI - Lupus update: perspective and clinical pearls. AB - Patients with systemic lupus erythematosus (SLE, lupus) have a markedly better survival rate today than they did 50 years ago, but they face a greater risk of cancer, cardiovascular disease, and osteoporosis at early ages. With better understanding of the immunological mechanisms of the disease, new avenues of therapy are emerging. PMID- 19188481 TI - Dehydrocostuslactone, a medicinal plant-derived sesquiterpene lactone, induces apoptosis coupled to endoplasmic reticulum stress in liver cancer cells. AB - This study is the first to investigate the anticancer effect of dehydrocostuslactone [DHE (3aS,6aR,9aR,9bS)-decahydro-3,6,9-tris(methylene) azuleno[4,5-b]furan-2(3H)-one)], a medicinal plant-derived sesquiterpene lactone, on hepatocellular carcinoma. Our results showed that DHE inhibits the proliferation of HepG2 and PLC/PRF/5 cells by inducing apoptosis. DHE induces up regulation of Bax and Bak, down-regulation of Bcl-2 and Bcl-XL, and nuclear relocation of the mitochondrial factors apoptosis-inducing factor (AIF) and endonuclease G (Endo G). DHE triggered endoplasmic reticulum (ER) stress, as indicated by changes in cytosol-calcium levels, double-stranded RNA-activated protein kinase-like endoplasmic reticulum kinase phosphorylation, inositol requiring protein 1 (IRE1) and CHOP/GADD153 up-regulation, X-box transcription factor-1 mRNA splicing, and caspase-4 activation. Enhancement of ER stress by DHE is through p38 and extracellular signal-regulated kinase 1/2-dependent manners and subsequently causes c-Jun NH(2)-terminal kinase activation, resulting in AIF and Endo G nuclear relocation. Both of IRE1 small interfering RNA transfection and 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester pretreatment inhibit DHE-mediated apoptosis, supporting the hypothesis that DHE induces cell death through ER stress. It is noteworthy that animal studies have revealed a dramatic 50% reduction in tumor volume after 45 days of treatment. This study demonstrates that DHE may be a novel anticancer agent for the treatment of liver cancer. PMID- 19188482 TI - Glycogen synthase kinase-3beta mediates endoplasmic reticulum stress-induced lysosomal apoptosis in leukemia. AB - Glycogen synthase kinase (GSK)-3beta may modulate endoplasmic reticulum (ER) stress-induced apoptosis; however, the mechanism remains unclear. Our data showed that human monocytic leukemia/lymphoma U937 and acute myeloid leukemia HL-60, but not chronic myeloid leukemia K562, cells were susceptible to apoptosis induced by ER stressor tunicamycin, a protein glycosylation inhibitor. Tunicamycin caused early activation of caspase-2, -3, -4, and -8, followed by apoptosis, whereas caspase-9 was slowly activated. Inhibiting caspase-2 reduced activation of caspase-8 and -3 but had no effect on caspase-4. Tunicamycin induced apoptosis independently of the mitochondrial pathway but caused lysosomal destabilization followed by lysosomal membrane permeabilization (LMP), cathepsin B relocation from lysosomes to the cytosol, and caspase-8 and -3 activation. It is notable that caspase-2 mediated lysosomal destabilization. Inhibiting GSK-3beta comprehensively reduced lysosomal apoptosis after caspase-2 inhibition. Unlike U937 and HL-60 cells, K562 cells showed nonresponsive ER stress and failure of activation of GSK-3beta and caspase-2 in response to tunicamycin. Activating GSK 3beta caused K562 cells to be susceptible to tunicamycin-induced apoptosis. Taken together, we show that GSK-3beta exhibits a mechanism of ER stress-induced lysosomal apoptosis in leukemia involving caspase-2-induced LMP and cathepsin B relocation, which result in caspase-8 and -3 activation. PMID- 19188483 TI - Colchicine: a novel positive allosteric modulator of the human 5 hydroxytryptamine3A receptor. AB - The actions of colchicine were examined with the two-electrode voltage-clamp technique and radioligand binding assays in mouse and human 5 hydroxytryptamine(3A) receptors (5-HT(3A)Rs) expressed in Xenopus laevis oocytes. Colchicine inhibited 5-hydroxytryptamine (5-HT)-evoked currents in oocytes expressing mouse 5-HT(3A)Rs, with an IC(50) of 59.5 +/- 3 microM. In contrast to the mouse receptor, coapplication of colchicine with 5-HT (<1 microM) strongly enhanced 5-HT-evoked currents in oocytes expressing human 5-HT(3A)Rs. Colchicine applied alone did not induce a detectable current. In the presence of 0.5 microM 5-HT, the potentiation was concentration-dependent and reached the maximum (approximately 100%) when 750 microM colchicine was applied. However, colchicine dependent inhibition can be observed at 5-HT concentrations > 1 microM. In oocyte membranes expressing mouse or human receptors, binding studies with colchicine (25 nM-1 mM) revealed no displacement of 1-methyl-N-((1R,3r,5S)-9-methyl-9 azabicyclo [3.3.1]nonan-3yl)-1H-indazole-3 carboxamide ([(3)H]BRL-43694), suggesting that actions of colchicine do not occur at the ligand binding domain. Functional effects of colchicine on both receptors occurred in the absence of preincubation and after cold temperature incubation, suggesting that the microtubule-depolymerizing effects of colchicine play no role in modulation of receptor function. Studies with interspecies chimeric receptors demonstrated that the distal one third of the N terminus is responsible for the bidirectional modulation by colchicine. Collectively, these results suggest that colchicine modulates receptor function through loops C and/or F through a gating mechanism. PMID- 19188484 TI - Migration inhibitory factor (MIF) released by macrophages upon recognition of immune complexes is critical to inflammation in Arthus reaction. AB - Deposition of immune complexes (IC) triggers Fc gamma R-dependent inflammation, leading to tissue damage in rheumatoid arthritis, systemic lupus erythematous, immune glomerulonephritis, and several immune vasculitides. Evidences support a role for macrophage migration inhibitory factor (MIF) in a number of inflammatory diseases, but the triggering of its secretion and its physiopathological role upon IC deposition remain elusive. Herein, we show that human macrophages secreted MIF after IC recognition, which in turn controlled the secretion of TNF. Macrophages from Mif-/- mice produced smaller amounts of TNF when stimulated with IgG-opsonized erythrocytes than wild-type (WT) cells. Using passive reverse Arthus reaction in the peritoneum and lungs as a model for IC-induced inflammation, we demonstrated that Mif-/- mice had a milder response, observed by reduced neutrophil recruitment, vascular leakage, and secretion of TNF, MIP-2, and keratinocyte-derived chemokine compared with WT controls. Adoptive transfer of alveolar macrophages from WT to Mif-/- mice rescued pulmonary neutrophil recruitment and TNF production upon passive reverse Arthus reaction. Our study indicates that Arthus inflammatory reaction is largely dependent on MIF and poses macrophages as a source of the MIF released upon IC recognition. These results give experimental support to the proposition that blockade of MIF might constitute an adjunctive, therapeutic approach to IC disease. PMID- 19188485 TI - Lipocalin2 expressions correlate significantly with tumor differentiation in epithelial ovarian cancer. AB - We recently identified lipocalin2 (LCN2) as being upregulated in ovarian cancer cell lines. The purpose of this study was to validate LCN2 upregulation in ovarian cancers and to investigate its potential as a serum biomarker. We assayed LCN2 expression in ovarian cancers using real-time PCR and IHC. To evaluate the potential of LCN2 as a biomarker, we measured serum LCN2 levels in 54 ovarian cancers, 15 borderline and 53 benign ovarian tumors, and 90 healthy controls. SYBR green PCR and IHC showed LCN2 overexpression in ovarian cancers. LCN2 immunoreactivity was significantly associated with tumor differentiation (p=0.009), as well-differentiated tumors showed the highest LCN2 expression. Serum LCN2 level in ovarian cancer was significantly higher than in the other study groups (p<0.001), and in accordance with IHC results, it also correlated with tumor differentiation, with well-differentiated tumors having the highest value. The sensitivity and specificity of LCN2 in detecting ovarian cancer was 72.2% and 50.4%, respectively. By Cox univariate analysis, LCN2 positivity was an independent prognostic factor for overall survival (hazard ratio = 1.47, p=0.012). In conclusion, LCN2 expressions are upregulated and related to tumor differentiation in ovarian cancers and should be included in future research assessing potential biomarkers for ovarian cancer. PMID- 19188486 TI - Eukaryotic initiation factors (eIF) 2alpha and 4E expression, localization, and phosphorylation in brain tumors. AB - Increased protein synthesis is regulated, in part, by two eukaryotic translation initiation factors (eIFs): eIF4E and eIF2alpha. One or both of these factors are often overexpressed in several types of cancer cells; however, no data are available at present regarding eIF4E and eIF2alpha levels in brain tumors. In this study, we analyzed the expression, subcellular localization and phosphorylation states of eIF4E and eIF2alpha in 64 brain tumors (26 meningiomas, 16 oligodendroglial tumors, and 22 astrocytomas) and investigated the correlation with the expression of MIB-1, p53, and cyclin D1 proteins as well. There are significant differences in the phosphorylated eIF4E levels between the tumors studied, being the highest in meningiomas and the lowest in the oligodendroglial tumors. Relative to subcellular localization, eIF4E is frequently found in the nucleus of the oligodendroglial tumors and rarely in the same compartment of the meningiomas, whereas eIF2alpha showed an inverse pattern. Finally, cyclin D1 levels directly correlate with the phosphorylation status of both factors. The different expression, phosphorylation, or/and subcellular distribution of eIF2alpha and eIF4E within the brain types of tumors studied could indicate that different pathways are activated for promoting cell cycle proliferation, for instance, leading to increased cyclin D1 expression. PMID- 19188487 TI - Mobilization of neural stem cells and generation of new neurons in 6-OHDA lesioned rats by intracerebroventricular infusion of liver growth factor. AB - Neural stem cells with self-renewal and multilineage potential persist in the subventricular zone of the adult mammalian forebrain. These cells remain relatively quiescent but, under certain conditions, can be stimulated, giving rise to new neurons. Liver growth factor (LGF) is a mitogen for liver cells that shows biological activity in extrahepatic sites and is useful for neuroregenerative therapies. The aim of this study was to investigate the potential neurogenic activity of LGF in the 6-hydroxydopamine rat model of Parkinson's disease. Proliferation was significantly increased in the subventricular zone and denervated striatum of rats receiving ICV LGF infusions, and 25% of the proliferating cells were doublecortin-positive neurons. Doublecortin-positive cells with the morphology of migrating neuroblasts were also observed in the dorsal and ventral regions of the striatum of LGF-infused animals. Moreover, some newly generated cells were neuronal nuclei-positive mature neurons. LGF also stimulated microglia and induced astrogliosis, both phenomena associated with generation and migration of new neurons in the adult brain. In summary, our study shows that LGF stimulates neurogenesis when applied intraventricularly in 6-hydroxydopamine-lesioned rats. Considering that this factor also promotes neuronal migration into damaged tissue, we propose LGF as a novel factor useful for neuronal replacement in neurodegenerative diseases. PMID- 19188488 TI - In situ zymography and immunolabeling in fixed and decalcified craniofacial tissues. AB - In situ zymography is a very important technique that shows the proteolytic activity in sections and allows researchers to observe the specific sites of proteolysis in tissues or cells. It is normally performed in non-fixed frozen sections and is not routinely performed in calcified tissues. In this study, we describe a technique that maintains proteolytic activity in fixed and decalcified sections obtained after routine paraffin sectioning in conventional microtome and cryostat sections. We used adult rat hemimandibles, which presented bone, enamel, and dentine matrices; the substrate used was dye-quenched-gelatin. Gelatinolytic activity was colocalized with MMP-2 using fluorescent antibodies. Specific proteolytic activity was observed in all sections, compatible with metalloproteinase activity, particularly in dentine and bone. Furthermore, matrix metalloproteinase-2 was colocalized to the sites of green fluorescence in dentine. In conclusion, the technique presented here will allow in situ zymography reactions in fixed, decalcified, and paraffin-embedded tissues, and we showed that paraformaldehyde-lysine-periodate-fixed cryostat sections are suitable for colocalization of gelatinolytic activity and protein labeling with antibodies. PMID- 19188489 TI - Enzyme activity and immunohistochemical localization of dipeptidyl peptidase 8 and 9 in male reproductive tissues. AB - The mRNA expression pattern of dipeptidyl peptidase (DPP) 8 and DPP9, two DPP4 homologs, was studied previously and showed a broad tissue distribution. In this study, protein expression and activity of DPP8 and DPP9 were investigated in male reproductive tissues of different mammals. Based on specific DPP activities and inhibition profiles, the proline-selective DPP activity in the bovine and rat testis could predominantly be attributed to DPP8/9 and not to DPP4. This is in contrast to the epididymis, where most of the activity was caused by DPP4. Bovine sperm preparations had very low or undetectable DPP8/9 activity. After characterization of polyclonal antibodies specific for DPP8 or DPP9, we could localize both enzymes in seminiferous tubules of the testis. A specific staining for DPP9 was found associated with spermatozoids embedded in the epithelium, just before their release into the lumen, and in spermatids. DPP8 was localized in spermatozoids in an earlier stage of maturation. These findings help to provide insight into the physiological role of DPP4-like enzymes in the male reproductive system. This manuscript contains online supplemental material at http://www.jhc.org. Please visit this article online to view these materials. PMID- 19188490 TI - The mitotic spindle mediates inheritance of the Golgi ribbon structure. AB - The mammalian Golgi ribbon disassembles during mitosis and reforms in both daughter cells after division. Mitotic Golgi membranes concentrate around the spindle poles, suggesting that the spindle may control Golgi partitioning. To test this, cells were induced to divide asymmetrically with the entire spindle segregated into only one daughter cell. A ribbon reforms in the nucleated karyoplasts, whereas the Golgi stacks in the cytoplasts are scattered. However, the scattered Golgi stacks are polarized and transport cargo. Microinjection of Golgi extract together with tubulin or incorporation of spindle materials rescues Golgi ribbon formation. Therefore, the factors required for postmitotic Golgi ribbon assembly are transferred by the spindle, but the constituents of functional stacks are partitioned independently, suggesting that Golgi inheritance is regulated by two distinct mechanisms. PMID- 19188491 TI - Mesenchymal cells reactivate Snail1 expression to drive three-dimensional invasion programs. AB - Epithelial-mesenchymal transition (EMT) is required for mesodermal differentiation during development. The zinc-finger transcription factor, Snail1, can trigger EMT and is sufficient to transcriptionally reprogram epithelial cells toward a mesenchymal phenotype during neoplasia and fibrosis. Whether Snail1 also regulates the behavior of terminally differentiated mesenchymal cells remains unexplored. Using a Snai1 conditional knockout model, we now identify Snail1 as a regulator of normal mesenchymal cell function. Snail1 expression in normal fibroblasts can be induced by agonists known to promote proliferation and invasion in vivo. When challenged within a tissue-like, three-dimensional extracellular matrix, Snail1-deficient fibroblasts exhibit global alterations in gene expression, which include defects in membrane type-1 matrix metalloproteinase (MT1-MMP)-dependent invasive activity. Snail1-deficient fibroblasts explanted atop the live chick chorioallantoic membrane lack tissue invasive potential and fail to induce angiogenesis. These findings establish key functions for the EMT regulator Snail1 after terminal differentiation of mesenchymal cells. PMID- 19188492 TI - Kinetochore stretching inactivates the spindle assembly checkpoint. AB - The spindle assembly checkpoint (SAC) monitors the attachment of microtubules to the kinetochore and inhibits anaphase when microtubule binding is incomplete. The SAC might also respond to tension; however, how cells can sense tension and whether its detection is important to satisfy the SAC remain controversial. We generated a HeLa cell line in which two components of the kinetochore, centromere protein A and Mis12, are labeled with green and red fluorophores, respectively. Live cell imaging of these cells reveals repetitive cycles of kinetochore extension and recoiling after biorientation. Under conditions in which kinetochore stretching is suppressed, cells fail to silence the SAC and enter anaphase after a delay, regardless of centromere stretching. Monitoring cyclin B levels as a readout for anaphase-promoting complex/cyclosome activity, we find that suppression of kinetochore stretching delays and decelerates cyclin B degradation. These observations suggest that the SAC monitors stretching of kinetochores rather than centromeres and that kinetochore stretching promotes silencing of the SAC signal. PMID- 19188493 TI - From fish to amphibians to mammals: in search of novel strategies to optimize cardiac regeneration. AB - Different vertebrate species have different cardiac regeneration rates: high in teleost fish, moderate in urodele amphibians, and almost negligible in mammals. Regeneration may occur through stem and progenitor cell differentiation or via dedifferentiation with residual cardiomyocytes reentering the cell cycle. In this review, we will examine the ability of zebrafish and newts to respond to cardiac damage with de novo cardiogenesis, whereas rodents and humans respond with a marked fibrogenic response and virtually no cardiomyocyte regeneration. Concerted strategies are needed to overcome this evolutionarily imposed barrier and optimize cardiac regeneration in mammals. PMID- 19188494 TI - Dendritic LSm1/CBP80-mRNPs mark the early steps of transport commitment and translational control. AB - Messenger RNA (mRNA) transport to neuronal dendrites is crucial for synaptic plasticity, but little is known of assembly or translational regulation of dendritic messenger ribonucleoproteins (mRNPs). Here we characterize a novel mRNP complex that is found in neuronal dendrites throughout the central nervous system and in some axonal processes of the spinal cord. The complex is characterized by the LSm1 protein, which so far has been implicated in mRNA degradation in nonneuronal cells. In brain, it associates with intact mRNAs. Interestingly, the LSm1-mRNPs contain the cap-binding protein CBP80 that associates with (pre)mRNAs in the nucleus, suggesting that the dendritic LSm1 complex has been assembled in the nucleus. In support of this notion, neuronal LSm1 is partially nuclear and inhibition of mRNA synthesis increases its nuclear localization. Importantly, CBP80 is also present in the dendrites and both LSm1 and CBP80 shift significantly into the spines upon stimulation of glutamergic receptors, suggesting that these mRNPs are translationally activated and contribute to the regulated local protein synthesis. PMID- 19188495 TI - Antagonistic regulation of Fus2p nuclear localization by pheromone signaling and the cell cycle. AB - When yeast cells sense mating pheromone, they undergo a characteristic response involving changes in transcription, cell cycle arrest in early G1, and polarization along the pheromone gradient. Cells in G2/M respond to pheromone at the transcriptional level but do not polarize or mate until G1. Fus2p, a key regulator of cell fusion, localizes to the tip of the mating projection during pheromone-induced G1 arrest. Although Fus2p was expressed in G2/M cells after pheromone induction, it accumulated in the nucleus until after cell division. As cells arrested in G1, Fus2p was exported from the nucleus and localized to the nascent tip. Phosphorylation of Fus2p by Fus3p was required for Fus2p export; cyclin/Cdc28p-dependent inhibition of Fus3p during late G1 through S phase was sufficient to block exit. However, during G2/M, when Fus3p was activated by pheromone signaling, Cdc28p activity again blocked Fus2p export. Our results indicate a novel mechanism by which pheromone-induced proteins are regulated during the transition from mitosis to conjugation. PMID- 19188496 TI - An essential role for p120-catenin in Src- and Rac1-mediated anchorage independent cell growth. AB - p120-catenin regulates epithelial cadherin stability and has been suggested to function as a tumor suppressor. In this study, we used anchorage-independent growth (AIG), a classical in vitro tumorigenicity assay, to examine the role of p120 in a different context, namely oncogene-mediated tumorigenesis. Surprisingly, p120 ablation by short hairpin RNA completely blocked AIG induced by both Rac1 and Src. This role for p120 was traced to its activity in suppression of the RhoA-ROCK pathway, which appears to be essential for AIG. Remarkably, the AIG block associated with p120 ablation was completely reversed by inhibition of the downstream RhoA effector ROCK. Harvey-Ras (H-Ras)-induced AIG was also dependent on suppression of the ROCK cascade but was p120 independent because its action on the pathway occurred downstream of p120. The data suggest that p120 modulates oncogenic signaling pathways important for AIG. Although H-Ras bypasses p120, a unifying theme for all three oncogenes is the requirement to suppress ROCK, which may act as a gatekeeper for the transition to anchorage independence. PMID- 19188497 TI - CD4+ regulatory T cells require CTLA-4 for the maintenance of systemic tolerance. AB - Cytotoxic T lymphocyte antigen-4 (CTLA-4) plays a critical role in negatively regulating T cell responses and has also been implicated in the development and function of natural FOXP3(+) regulatory T cells. CTLA-4-deficient mice develop fatal, early onset lymphoproliferative disease. However, chimeric mice containing both CTLA-4-deficient and -sufficient bone marrow (BM)-derived cells do not develop disease, indicating that CTLA-4 can act in trans to maintain T cell self tolerance. Using genetically mixed blastocyst and BM chimaeras as well as in vivo T cell transfer systems, we demonstrate that in vivo regulation of Ctla4(-/-) T cells in trans by CTLA-4-sufficient T cells is a reversible process that requires the persistent presence of FOXP3(+) regulatory T cells with a diverse TCR repertoire. Based on gene expression studies, the regulatory T cells do not appear to act directly on T cells, suggesting they may instead modulate the stimulatory activities of antigen-presenting cells. These results demonstrate that CTLA-4 is absolutely required for FOXP3(+) regulatory T cell function in vivo. PMID- 19188498 TI - The antigen-specific CD8+ T cell repertoire in unimmunized mice includes memory phenotype cells bearing markers of homeostatic expansion. AB - Memory T cells exhibit superior responses to pathogens and tumors compared with their naive counterparts. Memory is typically generated via an immune response to a foreign antigen, but functional memory T cells can also be produced from naive cells by homeostatic mechanisms. Using a recently developed method, we studied CD8 T cells, which are specific for model (ovalbumin) and viral (HSV, vaccinia) antigens, in unimmunized mice and found a subpopulation bearing markers of memory cells. Based on their phenotypic markers and by their presence in germ-free mice, these preexisting memory-like CD44(hi) CD8 T cells are likely to arise via physiological homeostatic proliferation rather than a response to environmental microbes. These antigen-inexperienced memory phenotype CD8 T cells display several functions that distinguish them from their CD44(lo) counterparts, including a rapid initiation of proliferation after T cell stimulation and rapid IFN-gamma production after exposure to proinflammatory cytokines. Collectively, these data indicate that the unprimed antigen-specific CD8 T cell repertoire contains antigen-inexperienced cells that display phenotypic and functional traits of memory cells. PMID- 19188499 TI - Down-regulation of Gfi-1 expression by TGF-beta is important for differentiation of Th17 and CD103+ inducible regulatory T cells. AB - Growth factor independent 1 (Gfi-1), a transcriptional repressor, is transiently induced during T cell activation. Interleukin (IL) 4 further induces Gfi-1, resulting in optimal Th2 cell expansion. We report a second important function of Gfi-1 in CD4 T cells: prevention of alternative differentiation by Th2 cells, and inhibition of differentiation of naive CD4 T cells to either Th17 or inducible regulatory T (iTreg) cells. In Gfi1(-/-) Th2 cells, the Rorc, Il23r, and Cd103 loci showed histone 3 lysine 4 trimethylation modifications that were lacking in wild-type Th2 cells, implying that Gfi-1 is critical for epigenetic regulation of Th17 and iTreg cell-related genes in Th2 cells. Enforced Gfi-1 expression inhibited IL-17 production and iTreg cell differentiation. Furthermore, a key inducer of both Th17 and iTreg cell differentiation, transforming growth factor beta, repressed Gfi-1 expression, implying a reciprocal negative regulation of CD4 T cell fate determination. Chromatin immunoprecipitation showed direct binding of the Gfi-1-lysine-specific demethylase 1 repressive complex to the intergenic region of Il17a/Il17f loci and to intron 1 of Cd103. T cell-specific Gfi1 conditional knockout mice displayed a striking delay in the onset of experimental allergic encephalitis correlated with a dramatic increase of Foxp3(+)CD103(+) CD4 T cells. Thus, Gfi-1 plays a critical role both in enhancing Th2 cell expansion and in repressing induction of Th17 and CD103(+) iTreg cells. PMID- 19188502 TI - Vascular endothelial growth factor-B induces myocardium-specific angiogenesis and arteriogenesis via vascular endothelial growth factor receptor-1- and neuropilin receptor-1-dependent mechanisms. AB - BACKGROUND: New revascularization therapies are urgently needed for patients with severe coronary heart disease who lack conventional treatment options. METHODS AND RESULTS: We describe a new proangiogenic approach for these no-option patients using adenoviral (Ad) intramyocardial vascular endothelial growth factor (VEGF)-B186 gene transfer, which induces myocardium-specific angiogenesis and arteriogenesis in pigs and rabbits. After acute infarction, AdVEGF-B186 increased blood vessel area, perfusion, ejection fraction, and collateral artery formation and induced changes toward an ischemia-resistant myocardial phenotype. Soluble VEGF receptor-1 and soluble neuropilin receptor-1 reduced the effects of AdVEGF B186, whereas neither soluble VEGF receptor-2 nor inhibition of nitric oxide production had this result. The effects of AdVEGF-B186 involved activation of neuropilin receptor-1, which is highly expressed in the myocardium, via recruitment of G-protein-alpha interacting protein, terminus C (GIPC) and upregulation of G-protein-alpha interacting protein. AdVEGF-B186 also induced an antiapoptotic gene expression profile in cardiomyocytes and had metabolic effects by inducing expression of fatty acid transport protein-4 and lipid and glycogen accumulation in the myocardium. CONCLUSIONS: VEGF-B186 displayed strikingly distinct effects compared with other VEGFs. These effects may be mediated at least in part via a G-protein signaling pathway. Tissue-specificity, high efficiency in ischemic myocardium, and induction of arteriogenesis and antiapoptotic and metabolic effects make AdVEGF-B186 a promising candidate for the treatment of myocardial ischemia. PMID- 19188501 TI - Intracellular calcium dynamics and acceleration of sinus rhythm by beta adrenergic stimulation. AB - BACKGROUND: Recent evidence indicates that membrane voltage and Ca2+ clocks jointly regulate sinoatrial node (SAN) automaticity. Here we test the hypothesis that sinus rate acceleration by beta-adrenergic stimulation involves synergistic interactions between these clock mechanisms. METHODS AND RESULTS: We simultaneously mapped intracellular calcium (Ca(i)) and membrane potential in 25 isolated canine right atrium, using previously described criteria of the timing of late diastolic Ca(i) elevation (LDCAE) relative to the action potential upstroke to detect the Ca2+ clock. Before isoproterenol, the earliest pacemaking site occurred in the inferior SAN, and LDCAE was observed in only 4 of 25 preparations. Isoproterenol infusion (1 micromol/L) increased sinus rate and shifted pacemaking site to superior SAN, concomitant with the appearance of LDCAE preceding the action potential upstroke by 98+/-31 ms. Caffeine had similar effects, whereas sarcoplasmic reticulum Ca2+ depletion with ryanodine and thapsigargin prevented isoproterenol-induced LDCAE and blunted sinus rate acceleration. Ca(i) transient relaxation time during isoproterenol was shorter in superior SAN (124+/-34 ms) than inferior SAN (138+/-24 ms; P=0.01) or right atrium (164+/-33 ms; P=0.001) and was associated with a lower sarcoplasmic reticulum Ca2+ ATPase pump to phospholamban protein ratio in SAN than in right atrium. Hyperpolarization-activated pacemaker current (I(f)) blockade with ZD 7288 modestly blunted but did not prevent LDCAE or sinus rate acceleration by isoproterenol. CONCLUSIONS: Acceleration of the Ca2+ clock in the superior SAN plays an important role in sinus acceleration during beta-adrenergic stimulation, interacting synergistically with the voltage clock to increase sinus rate. PMID- 19188504 TI - Cardiac Remote Ischemic Preconditioning in Coronary Stenting (CRISP Stent) Study: a prospective, randomized control trial. AB - BACKGROUND: Myocyte necrosis as a result of elective percutaneous coronary intervention (PCI) occurs in approximately one third of cases and is associated with subsequent cardiovascular events. This study assessed the ability of remote ischemic preconditioning (IPC) to attenuate cardiac troponin I (cTnI) release after elective PCI. METHODS AND RESULTS: Two hundred forty-two consecutive patients undergoing elective PCI with undetectable preprocedural cTnI were recruited. Subjects were randomized to receive remote IPC (induced by three 5 minute inflations of a blood pressure cuff to 200 mm Hg around the upper arm, followed by 5-minute intervals of reperfusion) or control (an uninflated cuff around the arm) before arrival in the catheter laboratory. The primary outcome was cTnI at 24 hours after PCI. Secondary outcomes included renal dysfunction and major adverse cardiac and cerebral event rate at 6 months. The median cTnI at 24 hours after PCI was lower in the remote IPC compared with the control group (0.06 versus 0.16 ng/mL; P=0.040). After remote IPC, cTnI was <0.04 ng/mL in 44 patients (42%) compared with 24 in the control group (24%; P=0.01). Subjects who received remote IPC experienced less chest discomfort (P=0.0006) and ECG ST segment deviation (P=0.005) than control subjects. At 6 months, the major adverse cardiac and cerebral event rate was lower in the remote IPC group (4 versus 13 events; P=0.018). CONCLUSIONS: Remote IPC reduces ischemic chest discomfort during PCI, attenuates procedure-related cTnI release, and appears to reduce subsequent cardiovascular events. PMID- 19188503 TI - AMP-activated protein kinase deficiency enhances myocardial ischemia/reperfusion injury but has minimal effect on the antioxidant/antinitrative protection of adiponectin. AB - BACKGROUND: Diabetes increases the morbidity/mortality of ischemic heart disease, but the underlying mechanisms are incompletely understood. Deficiency of both AMP activated protein kinase (AMPK) and adiponectin occurs in diabetes, but whether AMPK is cardioprotective or a central mediator of adiponectin cardioprotection in vivo remains unknown. METHODS AND RESULTS: Male adult mice with cardiomyocyte specific overexpression of a mutant AMPKalpha2 subunit (AMPK-DN) or wild-type (WT) littermates were subjected to in vivo myocardial ischemia/reperfusion (MI/R) and treated with vehicle or adiponectin. In comparison to WT, AMPK-DN mice subjected to MI/R endured greater cardiac injury (larger infarct size, more apoptosis, and poorer cardiac function) likely as a result of increased oxidative stress in these animals. Treatment of AMPK-DN mice with adiponectin failed to phosphorylate cardiac acetyl-CoA carboxylase as it did in WT mouse heart. However, a significant portion of the cardioprotection of adiponectin against MI/R injury was retained in AMPK-DN mice. Furthermore, treatment of AMPK-DN mice with adiponectin reduced MI/R-induced cardiac oxidative and nitrative stress to the same degree as that seen in WT mice. Finally, treating AMPK-DN cardiomyocytes with adiponectin reduced simulated MI/R-induced oxidative/nitrative stress and decreased cell death (P<0.01). CONCLUSIONS: Collectively, our results demonstrated that AMPK deficiency significantly increases MI/R injury in vivo but has minimal effect on the antioxidative/antinitrative protection of adiponectin. PMID- 19188506 TI - Comparison of early surgery versus conventional treatment in asymptomatic severe mitral regurgitation. AB - BACKGROUND: The optimal timing of surgical intervention in asymptomatic patients with severe mitral regurgitation is unclear. We therefore compared the long-term results of early surgery with a conventional treatment strategy. METHODS AND RESULTS: From 1996 to 2005, 447 consecutive asymptomatic patients (253 men, age 50+/-15 years) with severe degenerative mitral regurgitation and preserved left ventricular function were evaluated prospectively. The end point was defined as the composite of operative mortality, cardiac death, repeat mitral valve surgery, and urgent admission due to congestive heart failure during follow-up. Early surgery was performed on 161 patients (operated group), and the conventional treatment strategy was used for 286 patients (conventional treatment group). There were no significant differences between the 2 groups in terms of age, gender, euroSCORE (European System for Cardiac Operative Risk Evaluation), or ejection fraction. During a median follow-up of 1988 days, there were 2 repeat surgeries and no cardiac deaths or operative mortality in the operated group compared with 12 cardiac deaths, 1 repeat surgery, and 22 admissions for congestive heart failure in the conventional treatment group. The estimated actuarial 7-year cardiac mortality rate was 0% in the operated group and 5+/-2% in the conventional treatment group (P=0.008), and for 127 propensity score matched pairs, the estimated actuarial 7-year event-free survival rate was significantly higher in the operated than in the conventional treatment group (99+/-1% versus 85+/-4%, P=0.007). In the conventional treatment group, baseline grade of pulmonary hypertension (hazard ratio 1.87, 95% CI 1.22 to 2.87, P=0.003), age (hazard ratio 1.02, 95% CI 1.01 to 1.04, P=0.005), and effective regurgitant orifice area (hazard ratio 2.06, 95% CI 1.11 to 3.82, P=0.02) were independent variables that predicted late development of surgical indications or congestive heart failure on Cox multivariate analysis. CONCLUSIONS: Compared with conservative management, the strategy of early surgery was associated with an improved long-term event rate by decreasing cardiac mortality and congestive heart failure hospitalization more effectively in patients with severe degenerative mitral regurgitation. Early surgery may therefore further improve clinical outcomes in asymptomatic severe mitral regurgitation with preserved left ventricular systolic function and a high likelihood of mitral valve repair. PMID- 19188505 TI - Electrophysiological effects of late percutaneous coronary intervention for infarct-related coronary artery occlusion: the Occluded Artery Trial Electrophysiological Mechanisms (OAT-EP). AB - BACKGROUND: The Occluded Artery Trial-Electrophysiological Mechanisms (OAT-EP) tested the hypothesis that opening a persistently occluded infarct-related artery by percutaneous coronary intervention and stenting (PCI) after the acute phase of myocardial infarction compared with optimal medical therapy alone reduces markers of vulnerability to ventricular arrhythmias. METHODS AND RESULTS: Between April 2003 and December 2005, 300 patients with an occluded native infarct-related artery 3 to 28 days (median, 12 days) after myocardial infarction were randomized to PCI or optimal medical therapy. Ten-minute digital Holter recordings were obtained before randomization, at 30 days, and at 1 year. The primary end point was the change in alpha1, a nonlinear heart rate variability parameter, between baseline and 1 year. Major secondary end points were the changes in the filtered QRS duration on the signal-averaged ECG and variability in T-wave morphology (T wave variability) between baseline and 1 year. There were no significant differences in the changes in alpha1 (-0.04; 95% CI, -0.12 to 0.04), filtered QRS (2.2 ms; 95% CI, -1.4 to 5.9 ms), or T-wave variability (3.0 microV; 95% CI, -4.8 to 10.7 microV) between the PCI and medical therapy groups (medical therapy change minus PCI change). Multivariable analysis revealed that the results were unchanged after adjustment for baseline clinical variables and medication treatments during the Holter recordings. CONCLUSIONS: PCI with stenting of a persistently occluded infarct-related artery during the subacute phase after myocardial infarction compared with medical therapy alone had no significant effect on changes in heart rate variability, the time-domain signal-averaged ECG, or T-wave variability during the first year after myocardial infarction. These findings are consistent with the lack of clinical benefit, including no reduction in sudden death, with PCI for stable patients with persistently occluded infarct related arteries after myocardial infarction in the main OAT. PMID- 19188507 TI - Long-term clinical outcome after a first angiographically confirmed coronary stent thrombosis: an analysis of 431 cases. AB - BACKGROUND: There are limited data on the long-term clinical outcome after an angiographically confirmed (definite) stent thrombosis (ST). METHODS AND RESULTS: Four hundred thirty-one consecutive patients with a definite ST were enrolled in this multicenter registry. The primary end point was the composite of cardiac death and definite recurrent ST. Secondary end points were all-cause death, cardiac death, definite recurrent ST, definite and probable recurrent ST, any myocardial infarction, and any target-vessel revascularization. The primary end point occurred in 111 patients after a median follow-up of 27.1 months. The estimated cumulative event rates at 30 days and 1, 2, and 3 years were 18.0%, 23.6%, 25.2%, and 27.9%, respectively. The cumulative incidence rates of definite recurrent ST, definite or probable recurrent ST, any myocardial infarction, and any target-vessel revascularization were 18.8%, 20.1%, 21.3%, and 32.0%, respectively, at the longest available follow-up. Independent predictors for the primary end point were diabetes mellitus, total stent length, severe calcification, American College of Cardiology/American Heart Association B2-C lesions, TIMI (Thrombolysis In Myocardial Infarction) flow grade <3 after percutaneous coronary intervention, and left ventricular ejection fraction <45%. The implantation of an additional coronary stent during the first ST was also associated with unfavorable outcome. Clinical outcome was not affected by the type of previously implanted stent (drug-eluting or bare-metal stent) or the category of ST (early versus late). CONCLUSIONS: The long-term clinical outcome after a first definite ST is unfavorable, with a high mortality and recurrence rate. Diabetes mellitus, left ventricular ejection fraction <45%, long total stent length, complex coronary lesions, TIMI flow grade <3 after percutaneous coronary intervention, and implantation of an additional coronary stent during the emergent percutaneous coronary intervention for the ST were associated with this unfavorable outcome. PMID- 19188508 TI - Characteristics of children hospitalized with infective endocarditis. AB - BACKGROUND: Infective endocarditis in children is rare, and most reports describe the experience in referral centers. The purpose of our study was to assess the characteristics of children with infective endocarditis in a large national sample. METHODS AND RESULTS: We analyzed hospital discharge records with International Classification of Diseases, ninth revision, codes indicating infective endocarditis among admissions of patients <21 years of age in the Kids' Inpatient Databases 2000 and 2003; analyses for the 2 years were combined. In 1588 hospitalizations, the age distribution was bimodal, with peaks in infancy and late adolescence. The organism was coded in 632 admissions; Staphylococcus aureus was most common (57%), followed by the viridans group of streptococci (20%). Preexisting heart disease was present in 662 patients admitted (42%), among whom 81% had congenital heart disease, 8% had prosthetic valve endocarditis, and 5% had rheumatic heart disease. In-hospital mortality occurred in 84 patients (5%), 38 with preexisting heart disease. Death occurred in 12 of 25 patients (48%) with tetralogy of Fallot and pulmonary atresia, and 4 of 54 (8%) with prosthetic valve endocarditis. Among 46 deaths without preexisting heart disease, S aureus was the causative organism in 13 of 14 patients (93%) beyond infancy; among 32 infants who died, 10 (31%) were premature. CONCLUSIONS: In 2000 and 2003, we found a continuing shift in the epidemiology of pediatric infective endocarditis toward a higher proportion of children without preexisting heart disease. Risk factors for mortality included some forms of congenital heart disease and, among patients without preexisting heart disease, premature/neonatal age and S aureus as an etiologic agent. PMID- 19188509 TI - Diabetes mellitus, preexisting coronary heart disease, and the risk of subsequent coronary heart disease events in patients infected with human immunodeficiency virus: the Data Collection on Adverse Events of Anti-HIV Drugs (D:A:D Study). AB - BACKGROUND: Although guidelines in individuals not infected with the human immunodeficiency virus (HIV) consider diabetes mellitus (DM) to be a coronary heart disease (CHD) equivalent, there is little information on its association with CHD in those infected with HIV. We investigated the impact of DM and preexisting CHD on the development of a new CHD episode among 33,347 HIV-infected individuals in the Data Collection on Adverse Events of Anti-HIV Drugs (D:A:D Study). METHODS AND RESULTS: Over 159,971 person-years, 698 CHD events occurred. After adjustment for gender, age, cohort, HIV transmission, ethnicity, family history of CHD, smoking, and calendar year, the rate of a CHD episode was 7.52 times higher (Poisson regression, 95% CI 6.02 to 9.39, P=0.0001) in those with preexisting CHD than in those without preexisting CHD, but it was only 2.41 times higher (95% CI 1.91 to 3.05, P=0.0001) in those with preexisting DM compared with those without DM. No statistical interactions were apparent between either diagnosis and sex; although older people with DM had an increased CHD rate compared with younger people, older people with preexisting CHD had a lower event rate. A statistically significant interaction between preexisting DM and CHD (P=0.003) suggested that the CHD rate in those with preexisting CHD and DM is lower than expected on the basis of the main effects alone. CONCLUSIONS: DM and preexisting CHD are both important risk factors for CHD events in HIV-infected individuals. There is a need for targeted interventions to reduce the risk of CHD in both high-risk groups of HIV-infected individuals. PMID- 19188510 TI - Fasting blood glucose and the risk of stroke and myocardial infarction. AB - BACKGROUND: Although diabetes is a well-known risk factor of atherosclerotic cardiovascular diseases, the cardiovascular disease risk of glycemia below the current diabetic threshold remains uncertain. METHODS AND RESULTS: A total of 652,901 Korean men aged 30 to 64 years from the Korean National Health Insurance System were categorized into 8 groups by fasting blood glucose (FBG) level at baseline and were followed up for cardiovascular diseases occurrence during 1992 2001. Over the follow-up period of 8.8 years, 10,954 stroke and 3766 myocardial infarction events occurred. In age-adjusted analyses, there was evidence of linear associations between FBG and myocardial infarction, ischemic stroke, and intracerebral hemorrhagic stroke. However, with additional adjustment for socioeconomic position, behaviors, and other cardiovascular disease risk factors, the associations with myocardial infarction and intracerebral hemorrhagic stroke were markedly attenuated with increased risk only at the highest FBG levels (> or =7.5 mmol/L). With full adjustment, the association with ischemic stroke persisted; a linear increase in the risk of ischemic stroke was observed from FBG level of 5.6 mmol/L. When the analyses were repeated with those persons who had been diagnosed with diabetes removed, there was no evidence of associations of FBG with intracerebral hemorrhagic stroke, but the association with ischemic stroke persisted. CONCLUSIONS: In this Korean male population, the association with high FBG differed between ischemic stroke, intracerebral hemorrhagic stroke, and myocardial infarction. The linear increase in the risk of ischemic stroke, independently of other cardiovascular risk factors, was observed at a level below the current FBG criteria for impaired fasting glucose (> or =5.6 mmol/L). However, for other cardiovascular diseases, the current cutoff for diagnosing diabetes appropriately identified Korean men at risk. PMID- 19188511 TI - Mechanism of purinergic activation of endothelial nitric oxide synthase in endothelial cells. AB - BACKGROUND: Decreased endothelial nitric oxide (NO) synthase (eNOS) activity and NO production are critical contributors to the endothelial dysfunction and vascular complications observed in many diseases, including diabetes mellitus. Extracellular nucleotides activate eNOS and increase NO generation; however, the mechanism of this observation is not fully clarified. METHODS AND RESULTS: To elucidate the signaling pathway(s) leading to nucleotide-mediated eNOS phosphorylation at Ser-1177, human umbilical vein endothelial cells were treated with several nucleotides, including ATP, UTP, and ADP, in the presence or absence of selective inhibitors. These experiments identified P2Y1, P2Y2, and possibly P2Y4 as the purinergic receptors involved in eNOS phosphorylation and demonstrated that this process was adenosine independent. Nucleotide-induced eNOS phosphorylation and activity were inhibited by BAPTA-AM (an intracellular free calcium chelator), rottlerin (a protein kinase Cdelta inhibitor), and protein kinase Cdelta siRNA. In contrast, blockade of AMP-activated protein kinase, calcium/calmodulin-dependent kinase II, calcium/calmodulin-dependent kinase kinase, serine/threonine protein kinase B, protein kinase A, extracellular signal regulated kinase 1/2, and p38 mitogen-activated protein kinase did not affect nucleotide-mediated eNOS phosphorylation. CONCLUSIONS: The present study indicates that extracellular nucleotide-mediated eNOS phosphorylation is calcium and protein kinase Cdelta dependent. This newly identified signaling pathway opens new therapeutic avenues for the treatment of endothelial dysfunction. PMID- 19188513 TI - Smoking cessation early in pregnancy and birth weight, length, head circumference, and endothelial nitric oxide synthase activity in umbilical and chorionic vessels: an observational study of healthy singleton pregnancies. AB - BACKGROUND: Reduced production of the vasodilator nitric oxide (NO) in fetal vessels in pregnant smokers may lower the blood flow to the fetus and result in lower birth weight, length, and head circumference. The present study measured endothelial NO synthase (eNOS) activity in fetal umbilical and chorionic vessels from nonsmokers, smokers, and ex-smokers and related the findings to the fetal outcome. METHODS AND RESULTS: Of 266 healthy, singleton pregnancies, 182 women were nonsmokers, 43 were smokers, and 41 stopped smoking early in pregnancy. eNOS activity and concentration were quantified in endothelial cells of the fetal vessels. Cotinine, lipid profiles, estradiol, l-arginine, and dimethylarginines that may affect NO production were determined in maternal and fetal blood. Serum cotinine verified self-reported smoking. Newborns of smokers had a lower weight (P< or =0.001) and a smaller head circumference (P< or =0.041) and were shorter (P< or =0.001) than newborns of nonsmokers and ex-smokers. eNOS activity in umbilical veins of smokers was 36% lower (P<0.001), eNOS concentration was 47% lower (P<0.001), and the fetal plasma level of high-density lipoprotein was 18% lower (P<0.001) than those of nonsmokers, whereas the same levels were found in umbilical veins from ex-smokers and nonsmokers. The same patterns in eNOS activity and concentration were found in umbilical arteries and chorionic vessels. Fetal plasma levels of estradiol, l-arginine, dimethylarginines, total cholesterol, and triglycerides were similar for nonsmokers, smokers, and ex smokers. CONCLUSIONS: The findings suggest that maternal smoking reduces eNOS activity in the fetal vascular bed, contributing to retarded fetal growth caused by the reduction of vasodilatory capacity, and suggest that smoking cessation early in pregnancy prevents these effects in newborns. PMID- 19188512 TI - Ionizing radiation in cardiac imaging: a science advisory from the American Heart Association Committee on Cardiac Imaging of the Council on Clinical Cardiology and Committee on Cardiovascular Imaging and Intervention of the Council on Cardiovascular Radiology and Intervention. PMID- 19188514 TI - Images in Cardiovascular Medicine. Acute pericarditis assessed with magnetic resonance imaging: a new approach. PMID- 19188515 TI - Letter by ElBardissi regarding article, "Outcome of alcohol septal ablation for obstructive hypertrophic cardiomyopathy". PMID- 19188518 TI - Could dirty air cause diabetes? PMID- 19188517 TI - Severity of myocardial infarction: new insights on an elusive construct. PMID- 19188519 TI - Intracardiac echocardiography during interventional and electrophysiological cardiac catheterization. PMID- 19188520 TI - Optimizing trial design: sequential, adaptive, and enrichment strategies. PMID- 19188522 TI - Cardiovascular problems in pregnant women with marfan syndrome. PMID- 19188523 TI - Images in cardiovascular medicine. Anterior origin of the main pulmonary artery from the arterial valvar sinus: unusual truncus arteriosus. PMID- 19188521 TI - Prevention of atrial fibrillation: report from a national heart, lung, and blood institute workshop. AB - The National Heart, Lung, and Blood Institute convened an expert panel April 28 to 29, 2008, to identify gaps and recommend research strategies to prevent atrial fibrillation (AF). The panel reviewed the existing basic scientific, epidemiological, and clinical literature about AF and identified opportunities to advance AF prevention research. After discussion, the panel proposed the following recommendations: (1) enhance understanding of the epidemiology of AF in the population by systematically and longitudinally investigating symptomatic and asymptomatic AF in cohort studies; (2) improve detection of AF by evaluating the ability of existing and emerging methods and technologies to detect AF; (3) improve noninvasive modalities for identifying key components of cardiovascular remodeling that promote AF, including genetic, fibrotic, autonomic, structural, and electrical remodeling markers; (4) develop additional animal models reflective of the pathophysiology of human AF; (5) conduct secondary analyses of already-completed clinical trials to enhance knowledge of potentially effective methods to prevent AF and routinely include AF as an outcome in ongoing and future cardiovascular studies; and (6) conduct clinical studies focused on secondary prevention of AF recurrence, which would inform future primary prevention investigations. PMID- 19188524 TI - Transient receptor potential vanilloid type 4-deficient mice exhibit impaired endothelium-dependent relaxation induced by acetylcholine in vitro and in vivo. AB - Agonist-induced Ca2+ entry is important for the synthesis and release of vasoactive factors in endothelial cells. The transient receptor potential vanilloid type 4 (TRPV4) channel, a Ca2+-permeant cation channel, is expressed in endothelial cells and involved in the regulation of vascular tone. Here we investigated the role of TRPV4 channels in acetylcholine-induced vasodilation in vitro and in vivo using the TRPV4 knockout mouse model. The expression of TRPV4 mRNA and protein was detected in both conduit and resistance arteries from wild type mice. In small mesenteric arteries from wild-type mice, the TRPV4 activator 4alpha-phorbol-12,13-didecanoate increased endothelial [Ca2+]i in situ, which was reversed by the TRPV4 blocker ruthenium red. In wild-type animals, acetylcholine dilated small mesenteric arteries that involved both NO and endothelium-derived hyperpolarizing factors. In TRPV4-deficient mice, the NO component of the relaxation was attenuated and the endothelium-derived hyperpolarizing factor component was largely eliminated. Compared with their wild-type littermates, TRPV4-deficient mice demonstrated a blunted endothelial Ca2+ response to acetylcholine in mesenteric arteries and reduced NO release in carotid arteries. Acetylcholine (5 mg/kg, IV) decreased blood pressure by 37.0+/-6.2 mm Hg in wild type animals but only 16.6+/-2.7 mm Hg in knockout mice. We conclude that acetylcholine-induced endothelium-dependent vasodilation is reduced both in vitro and in vivo in TRPV4 knockout mice. These findings may provide novel insight into mechanisms of Ca2+ entry evoked by chemical agonists in endothelial cells. PMID- 19188525 TI - Systolic swing of the pendulum: relation between hypertension and heart failure revisited. PMID- 19188526 TI - Whites excrete a water load more rapidly than blacks. AB - A recent report demonstrated a racial difference in response to furosemide compatible with increased ion reabsorption in the thick ascending limb of the loop of Henle in blacks. Urinary dilution is another function of the loop diuretic-sensitive Na,K,2Cl cotransporter in the thick ascending limb, and racial differences in urinary diluting capacity have not been reported previously. We assessed diluting segment (cortical thick ascending limb and distal convoluted tubule) function in black and white normotensives in 2 studies using a water loading approach. In both studies, we found that whites excreted a water load more rapidly than blacks. In the first study, the final free water clearance rates (mean+/-SD) were 7.3+/-4.7 mL/min in whites (n=17, 7 females and 10 males) and 3.8+/-3.6 mL/min in blacks (n=14, 9 females and 5 males; P<0.03). In the second study, final free water clearance rates were 8.3+/-2.6 mL/min in whites (n=17, 8 females and 9 males) and 6.4+/-1.8 mL/min in blacks (n=11, 8 females and 3 males; P<0.01). We found no evidence of a racial difference in renal proximal tubular fluid reabsorption as assessed by renal endogenous lithium clearance or in plasma vasopressin level that could explain the difference in free water excretion. We conclude that our observations are most consistent with a lower capacity of ion reabsorption in the renal diluting segment in blacks. Slower excretion of an acute water load may have been an advantage during natural selection of humans living in arid, hot climates. PMID- 19188527 TI - Isolated systolic hypertension and incident heart failure in older adults: a propensity-matched study. AB - The association between isolated systolic hypertension (ISH) and incident heart failure (HF) has not been prospectively studied in a propensity-matched population of ambulatory older adults. Of the 5795 participants in the public-use copy of the Cardiovascular Health Study data set, 5248 had diastolic blood pressure <90 mm Hg and were free of HF at baseline. Of these, 2000 (38%) had ISH, defined as average seated systolic blood pressure > or =140 mm Hg. Propensity scores for baseline ISH were calculated for each participant (based on 64 baseline covariates) and were used to match 1260 pairs of participants with and without ISH. Matched Cox regression models were used to estimate the association of ISH with incident HF during a mean follow-up of 8.7 years. Matched participants (n=2520) had a mean (+/-SD) age of 74 (+/-6) years, 60% were women, 16% were nonwhites, 18% developed new-onset HF, and 35% died. Incident HF developed in 20% (rate: 242/10,000 person-years) and 16% (rate: 194/10,000 person years) of participants with and without ISH, respectively (matched hazard ratio when ISH was compared with no ISH: 1.26; 95% CI: 1.04 to 1.51; P=0.016). Prematch unadjusted, multivariable-adjusted, and propensity-adjusted hazard ratios (95% CIs) for ISH-associated incident HF were, respectively, 1.72 (1.51 to 1.97; P<0.0001), 1.35 (1.18 to 1.56; P<0.0001), and 1.22 (1.04 to 1.44; P=0.016). ISH had no association with all-cause mortality (matched hazard ratio: 1.03; 95% CI: 0.88 to 1.19; P=0.732). In conclusion, in a propensity-matched cohort of community-dwelling older adults who were well balanced in 64 baseline covariates, ISH was associated with increased risk of incident HF but had no association with all-cause mortality. PMID- 19188528 TI - Lifetime history of major depression predicts the development of the metabolic syndrome in middle-aged women. AB - OBJECTIVE: To prospectively examine the association of major depression with incidence of the metabolic syndrome in women. METHODS: Data were drawn from one of seven sites of the Study of Women's Health Across the Nation (SWAN), a prospective cohort study of the menopausal transition. Participants were 429 (34.5% African-American) women. Major depression and comorbid diagnoses were assessed via the Structured Clinical Interview for the Diagnostic and Statistical Manual of Mental Disorders, 4th Edition Axis I Disorders at baseline and seven annual follow-up evaluations. The metabolic syndrome was measured at baseline and each follow-up evaluation (except the second) based on National Cholesterol Education Program (NCEP) criteria. RESULTS: Longitudinal generalized estimating equations (GEE) models indicated that, in women who were free of the metabolic syndrome at baseline, a lifetime major depression history or current major depressive episode at baseline was significantly associated with the onset and presence of the metabolic syndrome during the follow-up (odds ratio = 1.82; 95% Confidence Interval (CI) = 1.06-3.14). Survival analyses showed that, in women who were free of the metabolic syndrome at baseline, a lifetime major depression history or current major depressive episode at baseline predicted increased risk of developing the metabolic syndrome during the follow-up (hazard ratio = 1.66; 95% CI = 0.99-3.75). Lifetime history of alcohol abuse or dependence predicted incident metabolic syndrome and attenuated the association between depression and the metabolic syndrome in both models. CONCLUSIONS: This study documents that major depression is a significant predictor of the onset of the metabolic syndrome. Intervention studies targeting depression may prevent the development of the metabolic syndrome in women. PMID- 19188529 TI - The Bypassing the Blues treatment protocol: stepped collaborative care for treating post-CABG depression. AB - OBJECTIVE: To present the design of the Bypassing the Blues (BtB) study to examine the impact of a collaborative care strategy for treating depression among patients with cardiac disease. Coronary artery bypass graft (CABG) surgery is one of the most common and costly medical procedures performed in the US. Up to half of post-CABG patients report depressive symptoms, and they are more likely to experience poorer health-related quality of life (HRQoL), worse functional status, continued chest pains, and higher risk of cardiovascular morbidity independent of cardiac status, medical comorbidity, and the extent of bypass surgery. METHODS: BtB was designed to enroll 450 post-CABG patients from eight Pittsburgh-area hospitals including: (1) 300 patients who expressed mood symptoms preceding discharge and at 2 weeks post hospitalization (Patient Health Questionnaire (PHQ-9) >or=10); and (2) 150 patients who served as nondepressed controls (PHQ-9 <5). Depressed patients were randomized to either an 8-month course of nurse-delivered telephone-based collaborative care supervised by a psychiatrist and primary care expert, or to their physicians' "usual care." The primary hypothesis will test whether the intervention can produce an effect size of >or=0.5 improvement in HRQoL at 8 months post CABG, as measured by the SF-36 Mental Component Summary score. Secondary hypotheses will examine the impact of our intervention on mood symptoms, cardiovascular morbidity, employment, health services utilization, and treatment costs. RESULTS: Not applicable. CONCLUSIONS: This effectiveness trial will provide crucial information on the impact of a widely generalizable evidence-based collaborative care strategy for treating depressed patients with cardiac disease. PMID- 19188531 TI - Associations of depression with C-reactive protein, IL-1, and IL-6: a meta analysis. AB - OBJECTIVE: To assess the magnitude and direction of associations of depression with C-reactive protein (CRP), interleukin (IL)-1, and IL-6 in community and clinical samples. METHODS: Systematic review of articles published between January 1967 and January 2008 in the PubMed and PsycINFO electronic databases was performed. Effect sizes were calculated as stat d and meta-analyzed, using random effects models. RESULTS: Each inflammatory marker was positively associated with depression; CRP, d = 0.15 (95% CI = 0.10, 0.21), p < .001; IL-6, d = 0.25 (95% CI = 0.18, 0.31), p < .001; IL-1, d = 0.35 (95% CI = 0.03, 0.67), p = .03; IL-1ra, d = 0.25 (95% CI = 0.04, 0.46), p = .02. Associations were strongest in clinically depressed patient samples--but were also significant in community-based samples- and when clinical interviews were used. Studies adjusting for body mass index (BMI) had smaller associations, albeit significant. Relationships were inconsistent with respect to age, medication, and sex. Depression was related to CRP and IL-6 among patients with cardiac disease or cancer. CONCLUSIONS: Depression and CRP, IL-1, and IL-6 are positively associated in clinical and community samples and BMI is implicated as a mediating/moderating factor. Continuity in clinic- and community-based samples suggests there is a dose response relationship between depression and these inflammatory markers, lending strength to the contention that the cardiac (or cancer) risk conferred by depression is not exclusive to patient populations. Available evidence is consistent with three causal pathways: depression to inflammation, inflammation to depression, and bidirectional relationships. PMID- 19188530 TI - Interactive effects of race and depressive symptoms on calcification in African American and white women. AB - OBJECTIVE: To examine the cross-sectional associations among race, depressive symptoms, and aortic and coronary calcification in a sample of middle-aged women. Depressive symptoms have been associated with atherosclerotic indicators of coronary heart disease (CHD) in white women. Few studies have examined these associations in samples including African American women, or explored whether any observed associations differ by race. METHODS: Participants were 508 (38% African American, 62% white) women. Aortic and coronary calcification were measured by electron beam tomography and depressive symptoms were assessed with the Center for Epidemiologic Studies Depression Scale (CES-D). Multivariable linear and logistic regression models were conducted to test associations. RESULTS: In linear regression models adjusted for race, depressive symptoms were associated with a greater amount of aortic calcification (beta = 0.03, p = .01), and there was a significant race x depressive symptoms interaction (beta = 0.07, p = .006). Findings for depressive symptoms (odds ratio (OR) = 1.03, 95% Confidence Interval (CI) = 1.0-1.06, p = .07), and the race x depressive symptoms interaction (OR = 1.1, 95% CI = 1.01-1.18, p = .01) were similar in race-adjusted multinomial logistic regression models predicting high levels of aortic calcification. Race specific models revealed a significant association between depressive symptoms and aortic calcification in African American, but not white women. Additional adjustments for education, study site, and CHD risk factors did not alter these results. Depressive symptoms were not associated with coronary calcification for women of either racial group. CONCLUSIONS: African American women may be particularly vulnerable to the effects of depressive symptoms on early atherosclerotic disease. PMID- 19188532 TI - Cumulative childhood stress and autoimmune diseases in adults. AB - OBJECTIVE: To examine whether childhood traumatic stress increased the risk of developing autoimmune diseases as an adult. METHODS: Retrospective cohort study of 15,357 adult health maintenance organization members enrolled in the Adverse Childhood Experiences (ACEs) Study from 1995 to 1997 in San Diego, California, and eligible for follow-up through 2005. ACEs included childhood physical, emotional, or sexual abuse; witnessing domestic violence; growing up with household substance abuse, mental illness, parental divorce, and/or an incarcerated household member. The total number of ACEs (ACE Score range = 0-8) was used as a measure of cumulative childhood stress. The outcome was hospitalizations for any of 21 selected autoimmune diseases and 4 immunopathology groupings: T- helper 1 (Th1) (e.g., idiopathic myocarditis); T-helper 2 (Th2) (e.g., myasthenia gravis); Th2 rheumatic (e.g., rheumatoid arthritis); and mixed Th1/Th2 (e.g., autoimmune hemolytic anemia). RESULTS: Sixty-four percent reported at least one ACE. The event rate (per 10,000 person-years) for a first hospitalization with any autoimmune disease was 31.4 in women and 34.4 in men. First hospitalizations for any autoimmune disease increased with increasing number of ACEs (p < .05). Compared with persons with no ACEs, persons with >or=2 ACEs were at a 70% increased risk for hospitalizations with Th1, 80% increased risk for Th2, and 100% increased risk for rheumatic diseases (p < .05). CONCLUSIONS: Childhood traumatic stress increased the likelihood of hospitalization with a diagnosed autoimmune disease decades into adulthood. These findings are consistent with recent biological studies on the impact of early life stress on subsequent inflammatory responses. PMID- 19188533 TI - Bernagchen Mahakala. PMID- 19188534 TI - Translating scientific opportunity into public health impact: a strategic plan for research on mental illness. AB - CONTEXT: Research has transformed many areas of medicine, with profound effects on morbidity and mortality. Exciting advances in neuroscience and genomics have transformed research but have not yet been translated to public health impact in psychiatry. Current treatments are necessary but not sufficient for most patients. OBJECTIVES: To improve outcomes we will need to (1) identify the neural circuitry of mental disorders, (2) detect the earliest manifestations of risk or illness even before cognition or behavior appear abnormal, (3) personalize care based on individual responses, and (4) implement broader use of effective psychosocial interventions. RESULTS: To address these objectives, NIMH, working with its many stakeholders, developed a strategic plan for research. The plan calls for research that will (1) define the pathophysiology of disorders from genes to behavior, (2) map the trajectory of illness to determine when, where, and how to intervene to preempt disability, (3) develop new interventions based on a personalized approach to the diverse needs and circumstances of people with mental illnesses, and (4) strengthen the public health impact of NIMH-supported research by focusing on dissemination science and disparities in care. CONCLUSIONS: The NIMH is shifting its funding priorities to close the gap between basic biological knowledge and effective mental health care, paving the way for prevention, recovery, and cure. PMID- 19188535 TI - Association of variants in DISC1 with psychosis-related traits in a large population cohort. AB - CONTEXT: There is an abundance of data from human genetic studies and animal models that implies a role for the disrupted in schizophrenia 1 gene (DISC1) in the etiology of schizophrenia and other major mental illnesses. OBJECTIVE: To study the effect of previously identified risk alleles of DISC1 on quantitative intermediate phenotypes for psychosis in an unselected population. DESIGN: We examined 41 single-nucleotide polymorphisms within DISC1 and performed tests of association with 4 quantitative phenotypes. SETTING: Academic research. PARTICIPANTS: Individuals from an unselected birth cohort in Finland. Originally, everyone born in the catchment area in 1966 (N = 12 058) was included in the study. Of these, 4651 (38.6%) attended the 31-year follow-up and could be included in the study. MAIN OUTCOME MEASURES: Scores on 4 psychometric instruments selected to function as proxies for positive and negative aspects of psychotic disorders, including the Perceptual Aberration Scale, Revised Social Anhedonia Scale, Revised Physical Anhedonia Scale, and Schizoidia Scale by Golden and Meehl. RESULTS: Carriers of the minor allele of marker rs821577 had significantly higher scores on social anhedonia (P < .001). The minor allele of marker rs821633 was strongly associated with lower scores on social anhedonia when analyzed dependent on the absence of the minor alleles of markers rs1538979 and rs821577 (P < .001). CONCLUSIONS: Variants in DISC1 affect the level of social anhedonia, a cardinal symptom of schizophrenia in the general population. DISC1 might be more central to human psychological functioning than previously thought, as it seems to affect the degree to which people enjoy social interactions. PMID- 19188536 TI - Influence of genes and environment on brain volumes in twin pairs concordant and discordant for bipolar disorder. AB - CONTEXT: Structural neuroimaging studies suggest the presence of subtle abnormalities in the brains of patients with bipolar disorder. The influence of genetic and/or environmental factors on these brain abnormalities is unknown. OBJECTIVE: To investigate the contribution of genetic and environmental factors on brain volume in bipolar disorder. DESIGN: Magnetic resonance imaging (1.5 T) brain scans of monozygotic (MZ) or dizygotic (DZ) twins concordant and discordant for bipolar disorder were compared with healthy twin pairs. SETTING: Subjects were recruited from the population, the Netherlands Twin Register, and the twin pair cohort at the University Medical Center Utrecht, Utrecht, The Netherlands. PARTICIPANTS: A total of 234 subjects including 50 affected twin pairs (9 MZ concordant; 15 MZ discordant; 4 DZ concordant; 22 DZ discordant) and 67 healthy twin pairs (39 MZ and 28 DZ) were included. MAIN OUTCOME MEASURES: Volumes of the intracranium, cerebrum, cerebellum, lateral and third ventricle, and gray and white matter from the cerebrum and frontal, parietal, temporal, and occipital lobes, both with and without correction for lithium use. To estimate the influence of additive genetic, common, and unique environmental factors, structural equation modeling was applied. RESULTS: Bipolar disorder was associated with a decrease in total cortical volume. Decreases in white matter were related to the genetic risk of developing bipolar disorder (bivariate heritability, 77%; 95% confidence interval, 38% to 100%). Significant environmental correlations were found for cortical gray matter. These relationships all became more pronounced when data were corrected for lithium use. CONCLUSIONS: Focusing on genes controlling white matter integrity may be a fruitful strategy in the quest to discover genes implicated in bipolar disorder. Elucidating the mechanism by which lithium attenuates brain matter loss may lead to the development of neuroprotective drugs. PMID- 19188537 TI - The intricate link between violence and mental disorder: results from the National Epidemiologic Survey on Alcohol and Related Conditions. AB - CONTEXT: The relationship between mental illness and violence has a significant effect on mental health policy, clinical practice, and public opinion about the dangerousness of people with psychiatric disorders. OBJECTIVE: To use a longitudinal data set representative of the US population to clarify whether or how severe mental illnesses such as schizophrenia, bipolar disorder, and major depression lead to violent behavior. DESIGN: Data on mental disorder and violence were collected as part of the National Epidemiologic Survey on Alcohol and Related Conditions (NESARC), a 2-wave face-to-face survey conducted by the National Institute on Alcohol Abuse and Alcoholism. PARTICIPANTS: A total of 34 653 subjects completed NESARC waves 1 (2001-2003) and 2 (2004-2005) interviews. Wave 1 data on severe mental illness and risk factors were analyzed to predict wave 2 data on violent behavior. MAIN OUTCOME MEASURES: Reported violent acts committed between waves 1 and 2. RESULTS: Bivariate analyses showed that the incidence of violence was higher for people with severe mental illness, but only significantly so for those with co-occurring substance abuse and/or dependence. Multivariate analyses revealed that severe mental illness alone did not predict future violence; it was associated instead with historical (past violence, juvenile detention, physical abuse, parental arrest record), clinical (substance abuse, perceived threats), dispositional (age, sex, income), and contextual (recent divorce, unemployment, victimization) factors. Most of these factors were endorsed more often by subjects with severe mental illness. CONCLUSIONS: Because severe mental illness did not independently predict future violent behavior, these findings challenge perceptions that mental illness is a leading cause of violence in the general population. Still, people with mental illness did report violence more often, largely because they showed other factors associated with violence. Consequently, understanding the link between violent acts and mental disorder requires consideration of its association with other variables such as substance abuse, environmental stressors, and history of violence. PMID- 19188538 TI - Risk of postpartum depressive symptoms with elevated corticotropin-releasing hormone in human pregnancy. AB - CONTEXT: Postpartum depression (PPD) is common and has serious implications for the mother and her newborn infant. A possible link between placental corticotropin-releasing hormone (pCRH) and PPD incidence has been hypothesized, but empirical evidence is lacking. OBJECTIVE: To determine whether accelerated increases in pCRH throughout pregnancy are associated with PPD symptoms. DESIGN: Pregnant women were recruited into this longitudinal cohort study. Blood samples were obtained at 15, 19, 25, 31, and 37 weeks' gestational age (GA) for assessment of pCRH, cortisol, and adrenocorticotropic hormone (ACTH). Depressive symptoms were assessed with a standardized questionnaire at the last 4 pregnancy visits and post partum. SETTING: Subjects were recruited from 2 southern California medical centers, and visits were conducted in research laboratories. PARTICIPANTS: One hundred adult women with a singleton pregnancy. Main Outcome Measure Symptoms of PPD were assessed at a mean (SD) of 8.7 (2.94) weeks after delivery with the Edinburgh Postnatal Depression Scale. RESULTS: Sixteen women developed PPD symptoms. At 25 weeks' GA, pCRH was a strong predictor of PPD symptoms (R(2) = 0.21; beta = 0.46 [P < .001]), an effect that remained significant after controlling for prenatal depressive symptoms. No significant associations were found for cortisol and ACTH. Receiver operating characteristic curve analyses revealed that pCRH at 25 weeks' GA is a possible diagnostic tool (area under the curve, 0.78 [P = .001]). Sensitivity (0.75) and specificity (0.74) at the ideal cutoff point (pCRH, 56.86 pg/mL) were moderate. Growth curve analyses indicated that the trajectories of pCRH in women with PPD symptoms are significantly accelerated from 23 to 26 weeks' GA. CONCLUSIONS: At a critical period in midpregnancy, pCRH is a sensitive and specific early diagnostic test for PPD symptoms. If replicated, these results have implications for the identification and treatment of pregnant women at risk for PPD. PMID- 19188539 TI - Neural bases of social anxiety disorder: emotional reactivity and cognitive regulation during social and physical threat. AB - CONTEXT: Social anxiety disorder is thought to involve emotional hyperreactivity, cognitive distortions, and ineffective emotion regulation. While the neural bases of emotional reactivity to social stimuli have been described, the neural bases of emotional reactivity and cognitive regulation during social and physical threat, and their relationship to social anxiety symptom severity, have yet to be investigated. OBJECTIVE: To investigate behavioral and neural correlates of emotional reactivity and cognitive regulation in patients and controls during processing of social and physical threat stimuli. DESIGN: Participants were trained to implement cognitive-linguistic regulation of emotional reactivity induced by social (harsh facial expressions) and physical (violent scenes) threat while undergoing functional magnetic resonance imaging and providing behavioral ratings of negative emotion experience. SETTING: Academic psychology department. PARTICIPANTS: Fifteen adults with social anxiety disorder and 17 demographically matched healthy controls. MAIN OUTCOME MEASURES: Blood oxygen level-dependent signal and negative emotion ratings. RESULTS: Behaviorally, patients reported greater negative emotion than controls during social and physical threat but showed equivalent reduction in negative emotion following cognitive regulation. Neurally, viewing social threat resulted in greater emotion-related neural responses in patients than controls, with social anxiety symptom severity related to activity in a network of emotion- and attention-processing regions in patients only. Viewing physical threat produced no between-group differences. Regulation during social threat resulted in greater cognitive and attention regulation related brain activation in controls compared with patients. Regulation during physical threat produced greater cognitive control-related response (ie, right dorsolateral prefrontal cortex) in patients compared with controls. CONCLUSIONS: Compared with controls, patients demonstrated exaggerated negative emotion reactivity and reduced cognitive regulation-related neural activation, specifically for social threat stimuli. These findings help to elucidate potential neural mechanisms of emotion regulation that might serve as biomarkers for interventions for social anxiety disorder. PMID- 19188540 TI - Association between media use in adolescence and depression in young adulthood: a longitudinal study. AB - CONTEXT: Although certain media exposures have been linked to the presence of psychiatric conditions, few studies have investigated the association between media exposure and depression. OBJECTIVE: To assess the longitudinal association between media exposure in adolescence and depression in young adulthood in a nationally representative sample. DESIGN: Longitudinal cohort study. SETTING AND PARTICIPANTS: We used the National Longitudinal Survey of Adolescent Health (Add Health) to investigate the relationship between electronic media exposure in 4142 adolescents who were not depressed at baseline and subsequent development of depression after 7 years of follow-up. Main Outcome Measure Depression at follow up assessed using the 9-item Center for Epidemiologic Studies-Depression Scale. RESULTS: Of the 4142 participants (47.5% female and 67.0% white) who were not depressed at baseline and who underwent follow-up assessment, 308 (7.4%) reported symptoms consistent with depression at follow-up. Controlling for all covariates including baseline Center for Epidemiologic Studies-Depression Scale score, those reporting more television use had significantly greater odds of developing depression (odds ratio [95% confidence interval], 1.08 [1.01-1.16]) for each additional hour of daily television use. In addition, those reporting more total media exposure had significantly greater odds of developing depression (1.05 [1.0004-1.10]) for each additional hour of daily use. We did not find a consistent relationship between development of depressive symptoms and exposure to videocassettes, computer games, or radio. Compared with young men, young women were less likely to develop depression given the same total media exposure (odds ratio for interaction term, 0.93 [0.88-0.99]). CONCLUSION: Television exposure and total media exposure in adolescence are associated with increased odds of depressive symptoms in young adulthood, especially in young men. PMID- 19188541 TI - Risks and predictors of readmission for a mental disorder during the postpartum period. AB - CONTEXT: It has been suggested that the risk of inpatient psychiatric readmissions is elevated during the postpartum period. To our knowledge, no prior study has compared mothers and nonmothers to determine whether the risk of readmission differs between these 2 groups of women. OBJECTIVES: To compare mothers and nonmothers to assess whether childbirth increases the risk for psychiatric readmission and to identify predictors of psychiatric readmission during the postpartum period. DESIGN: A population-based cohort study merging data from the Danish Civil Registration System and the Danish Psychiatric Central Register. SETTING: The population of Denmark. PARTICIPANTS: Two partly overlapping study populations included a total of 28 124 women, 10 218 of whom were mothers, who were followed up from January 1, 1973, through June 30, 2005. Main Outcome Measure Readmission rates to psychiatric hospitals during the 12 months after childbirth (first live-born child). RESULTS: The period of highest risk of psychiatric readmission in new mothers was 10 to 19 days post partum (relative risk [RR], 2.71; 95% confidence interval [CI], 1.68-4.37), and the period of lowest risk was during pregnancy (0.54; 0.43-0.69). Childbirth was associated with an increased risk of readmission during the first postpartum month, after which risk for readmission was higher among nonmothers (RR, 1.53; 95% CI, 1.31-1.80). A previous diagnosis of bipolar affective disorder was the strongest predictor of readmissions 10 to 19 days post partum (RR, 37.22; 95% CI, 13.58-102.04). In all, 26.9% of mothers with this diagnosis were readmitted within the first postpartum year. CONCLUSIONS: Mothers with mental disorders have lower readmission rates compared with women with mental disorders who do not have children. However, the first month after childbirth is associated with increased risk of psychiatric readmission, and women with a history of bipolar affective disorder are at particular risk of postpartum psychiatric readmissions. PMID- 19188542 TI - Relationship of type 1 cannabinoid receptor availability in the human brain to novelty-seeking temperament. AB - CONTEXT: Brain neurochemistry can partially account for personality traits as a variance of normal human behavior, as has been demonstrated for monoamine neurotransmission. Positron emission tomography using fluorine 18-labeled MK-9470 now enables quantification of type 1 cannabinoid receptors (CB1R) in the brain. OBJECTIVE: To investigate whether there is a relationship between human temperament traits and regional cerebral CB1R availability. DESIGN: Forty-seven [(18)F]MK-9470 baseline scanning sessions were performed and correlated with the temperament dimensions and subdimensions of the 240-item Cloninger Temperament and Character Inventory. SETTING: Academic brain imaging center. PARTICIPANTS: Forty-seven nonsmoking, healthy volunteers (paid). Main Outcome Measure Voxel based correlation of temperament variables of the inventory with regional CB1R availability. RESULTS: Novelty seeking was inversely correlated with global CB1R availability (r = -0.33, P = .02), with the most significant correlation in the left amygdala (r = -0.41, P = .005). In particular, the subdimension extravagance showed a highly significant inverse correlation to global CB1R availability (r = 0.53, P <.001), most pronounced in the amygdala, anterior cingulate, parietal cortex, and precuneus. Also, disorderliness was inversely correlated with global CB1R availability (r = -0.31, P = .04). CONCLUSIONS: Low baseline cerebral CB1R availability is related to a high novelty-seeking personality, in particular to extravagance, most pronounced in the amygdala. Further investigation of the functional role of the CB1R is warranted in pathological behavior known to be strongly related to novelty seeking, such as addiction and eating disorders. PMID- 19188543 TI - Responsiveness to drug cues and natural rewards in opiate addiction: associations with later heroin use. AB - CONTEXT: Although drug cues reliably activate the brain's reward system, studies rarely examine how the processing of drug stimuli compares with natural reinforcers or relates to clinical outcomes. OBJECTIVES: To determine hedonic responses to natural and drug reinforcers in long-term heroin users and to examine the utility of these responses in predicting future heroin use. DESIGN: Prospective design examining experiential, expressive, reflex modulation, and cortical/attentional responses to opiate-related and affective stimuli. The opiate-dependent group was reassessed a median of 6 months after testing to determine their level of heroin use during the intervening period. SETTING: Community drug and alcohol services and a clinical research facility. PARTICIPANTS: Thirty-three opiate-dependent individuals (mean age, 31.6 years) with stabilized opiate-substitution pharmacotherapy and 19 sex- and age-matched healthy non-drug users (mean age, 30 years). MAIN OUTCOME MEASURES: Self-ratings, facial electromyography, startle-elicited postauricular reflex, and event-related potentials combined with measures of heroin use at baseline and follow-up. RESULTS: Relative to the control group, the opiate-dependent group rated pleasant pictures as less arousing and showed increased corrugator activity, less postauricular potentiation, and decreased startle-elicited P300 attenuation while viewing pleasant pictures. The opiate-dependent group rated the drug-related pictures as more pleasant and arousing, and demonstrated greater startle-elicited P300 attenuation while viewing them. Although a startle-elicited P300 amplitude response to pleasant (relative to drug-related) pictures significantly predicted regular (at least weekly) heroin use at follow-up, subjective valence ratings of pleasant pictures remained the superior predictor of use after controlling for baseline craving and heroin use. CONCLUSIONS: Heroin users demonstrated reduced responsiveness to natural reinforcers across a range of psychophysiological measures. Subjective rating of pleasant pictures robustly predicted future heroin use. Our findings highlight the importance of targeting anhedonic symptoms within clinical treatment settings. PMID- 19188544 TI - Racial differences in visit duration of outpatient psychiatric visits. AB - CONTEXT: Substantial racial disparities exist in the delivery of some health care services. Whether racial disparities exist in the duration of office visits to psychiatrists is not known. OBJECTIVE: To compare the duration of visits to office-based psychiatrists by white and African American patients. DESIGN, SETTING, AND PARTICIPANTS: Analysis of a nationally representative sample of visits to office-based psychiatrists between 2001 and 2006. Visits were grouped by patient race as non-Hispanic African American (n = 504) or non-Hispanic white (n = 7094). Main Outcome Measure Duration of face-to-face contact between patient and psychiatrist. RESULTS: Unadjusted mean duration of psychiatric outpatient visits by African Americans (mean duration, 28.3 minutes) were 4.4 minutes shorter than visits by whites (32.7 minutes) (P = .02), although the difference narrowed (3.5 minutes; P = .07) following adjustment for potentially confounding patient, psychiatrist, and practice characteristics. A gap was evident in 2001 2003 (7.4 minutes; P <.001) but negligible in 2004-2006 (0.1 minute; P = .94). In stratified regressions that combined time periods and controlled for several relevant characteristics, significant racial differences in visit duration were observed among visits with the following characteristics: adjustment disorder diagnosis (10.0 minutes; P <.001), female patient sex (5.4 minutes; P = .008), depressive disorder diagnosis (5.2 minutes; P = .04), solo practice (5.2 minutes; P = .04), psychotherapy provision (5.1 minutes; P = .01), practices with high patient volume (5.0 minutes; P = .03), Medicare payment (3.5 minutes; P = .02), and absence of psychiatric comorbidity (3.3 minutes; P = .04). CONCLUSIONS: In recent years, progress has been made in closing a racial gap in the length of psychiatric office-based outpatient visits by African American vs white patients. Against a backdrop of persisting racial disparities in other areas of mental health care, ongoing attention to reducing disparities will be necessary to sustain and extend these gains. PMID- 19188545 TI - Impact of preoperative chemotherapy on pulmonary function tests in resectable early-stage non-small cell lung cancer. AB - BACKGROUND: Several chemotherapy agents, including gemcitabine and paclitaxel, have been reported to cause interstitial pneumonitis. The incidence of pulmonary toxicity from the combination of gemcitabine and paclitaxel is reported to be approximately 5%. In this report, pulmonary function test (PFT) results were analyzed from two similar randomized phase 2 trials that tested platinum and nonplatinum regimens preoperatively in patients with stage I or II non-small cell lung cancer (NSCLC). METHODS: The regimens included gemcitabine plus carboplatin, paclitaxel, or cisplatin. PFT and dyspnea scores were obtained at baseline and postchemotherapy, and were compared to one of several secondary end points, including ability to undergo surgical resection. RESULTS: Baseline PFT scores varied with smoking status. Mean levels of diffusing capacity of the lung for carbon monoxide (Dlco) adjusted for hemoglobin declined 8% from pre- to postinduction (Wilcoxon signed rank test, p < 0.0001). Changes in FVC, FEV(1), and total lung capacity were not statistically significant after chemotherapy. Although 27% of patients in the study had some reduction in PFT results, only 2 of the 85 eligible patients did not undergo surgery due to PFT reduction following chemotherapy. One patient in the study experienced a clinically significant respiratory toxicity (grade 3 dyspnea). Pulmonary toxicity was only statistically associated with male gender. CONCLUSION: In the preoperative setting, gemcitabine-based chemotherapy was well tolerated. The most commonly affected PFT parameter postchemotherapy was the Dlco. Although 15% of patients had a significant reduction in the Dlco postchemotherapy, it did not correlate with clinical symptoms or affect the ability to undergo surgical resection. PMID- 19188546 TI - American College of Chest Physicians/La Societe de Reanimation de Langue Francaise statement on competence in critical care ultrasonography. AB - OBJECTIVE: To define competence in critical care ultrasonography (CCUS). DESIGN: The statement is sponsored by the Critical Care NetWork of the American College of Chest Physicians (ACCP) in partnership with La Societe de Reanimation de Langue Francaise (SRLF). The ACCP and the SRLF selected a panel of experts to review the field of CCUS and to develop a consensus statement on competence in CCUS. RESULTS: CCUS may be divided into general CCUS (thoracic, abdominal, and vascular), and echocardiography (basic and advanced). For each component part, the panel defined the specific skills that the intensivist should have to be competent in that aspect of CCUS. CONCLUSION: In defining a reasonable minimum standard for CCUS, the statement serves as a guide for the intensivist to follow in achieving proficiency in the field. PMID- 19188547 TI - A randomized controlled trial of standard vs endobronchial ultrasonography-guided transbronchial needle aspiration in patients with suspected sarcoidosis. AB - BACKGROUND: Endobronchial ultrasound (EBUS)-guided transbronchial needle aspiration (TBNA) of mediastinal lymph nodes has been found to be more accurate than standard TBNA in the setting of malignancy. In patients with suspected sarcoidosis, the smaller ultrasound needle may yield inadequate material to make a histologic diagnosis of granulomatous inflammation. The aim of this study was to compare the diagnostic yield of EBUS-guided TBNA to TBNA performed with a standard 19-gauge needle in patients with mediastinal adenopathy and a clinical suspicion of sarcoidosis. METHODS: A randomized controlled trial was performed in a university medical center, enrolling 50 patients (of 61 screened, 2 declined, and 9 did not meet entry criteria) with hilar and/or mediastinal adenopathy and a clinical suspicion of sarcoidosis. Twenty-four patients were randomized to undergo EBUS-guided TBNA and 26 to undergo TBNA using a standard 19-gauge needle. RESULTS: The primary outcome measure of diagnostic yield was 53.8% vs 83.3% in favor of the EBUS-guided TBNA group, an absolute increase of 29.5% (p < 0.05; 95% confidence interval [CI], 8.6 to 55.4%). After blinded research pathology review, diagnostic yield was 73.1% vs 95.8%, in favor of the EBUS-guided TBNA group, an absolute increase of 22.7% (p = 0.05; 95% CI, 1.9 to 42.2%). Sensitivity and specificity were 60.9% and 100%, respectively, in the standard TBNA group, and 83.3% and 100%, respectively, in the EBUS-guided TBNA group (absolute increase in sensitivity, 22.5%; p = 0.085; 95% CI, 3.2 to 44.9%). CONCLUSIONS: The diagnostic yield of EBUS-guided TBNA is superior to TBNA using a standard 19-gauge needle for sampling of mediastinal lymph nodes in patients with a clinical suspicion of sarcoidosis. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT00373555. PMID- 19188548 TI - Failure of medical therapy in pulmonary arterial hypertension. Is there an alternative diagnosis? AB - BACKGROUND: Despite the development of targeted therapies for pulmonary arterial hypertension (PAH), some patients fail to respond to medical therapy. There are a number of types of PAH, one of which is pulmonary venoocclusive disease (PVOD). Unlike other PAH types, the hallmark pathology of PVOD is fibrous occlusion of the small venules, potentially with arterial involvement. It was hypothesized that a lack of response to medical therapy in clinically diagnosed PAH can be explained by misdiagnosed PVOD. METHODS: This study reviewed cases of 14 patients with clinically diagnosed PAH who had failed to respond to medical therapy and had lung tissue available from autopsy or explant. Control samples (n = 6) were obtained from lungs explanted for other causes, and a previous transthoracic echocardiogram excluded pulmonary hypertension. Comprehensive vessel morphometry was performed, and the clinical data reviewed. RESULTS: Vessel analysis confirmed 86% of case patients had PVOD and 14% had arterial-only PAH (artPAH). In the PVOD case patients, significant pathology was present in all vessel types with similar obstructive, intimal fibrosis in the venules (p < 0.0001) and arterioles (p < 0.0001). This was considerably different from the intimal concentric laminar smooth muscle proliferation in the artPAH cases. There were no significant differences in clinical presentation between PVOD and artPAH case patients. Pulmonary edema was not observed in the PVOD cases. CONCLUSIONS: PVOD is an underdiagnosed and commonly misdiagnosed cause of pulmonary hypertension, which may present with a failure to respond to medical therapy. Further studies on PVOD may help confirm whether a change in its classification as a type of PAH is necessary. PMID- 19188549 TI - Identification of early acute lung injury at initial evaluation in an acute care setting prior to the onset of respiratory failure. AB - BACKGROUND: Despite being a focus of intensive investigation, acute lung injury (ALI) remains a major cause of morbidity and mortality. However, the current consensus definition impedes identification of patients with ALI before they require mechanical ventilation. To establish a definition of early ALI (EALI), we carried out a prospective cohort study to identify clinical predictors of progression to ALI. METHODS: Potential cases of EALI were identified by daily screening of chest radiographs (CXRs) for all adult emergency department and new medicine service admissions at Stanford University Hospital. RESULTS: Of 1,935 screened patients with abnormal CXRs, we enrolled 100 patients admitted with bilateral opacities present < 7 days and not due exclusively to left atrial hypertension. A total of 33 of these 100 patients progressed to ALI requiring mechanical ventilation during their hospitalization. Progression to ALI was associated with immunosuppression, the modified Rapid Emergency Medicine Score, airspace opacities beyond the bases, systemic inflammatory response syndrome, and the initial oxygen requirement (> 2 L/min). On multivariate analysis, only an initial oxygen requirement > 2 L/min predicted progression to ALI (odds ratio, 8.1; 95% confidence interval, 2.7 to 24.5). A clinical diagnosis of EALI, defined by hospital admission with bilateral opacities on CXR not exclusively due to left atrial hypertension and an initial oxygen requirement of > 2 L/min, was 73% sensitive and 79% specific for progression to ALI. CONCLUSIONS: A new clinical definition of EALI may have value in identifying patients with ALI early in their disease course. PMID- 19188550 TI - Bosentan decreases pulmonary vascular resistance and improves exercise capacity in acute hypoxia. AB - BACKGROUND: Altitude exposure is associated with mild pulmonary hypertension and decreased exercise capacity. We tested the hypothesis that pulmonary vascular resistance (PVR) contributes to decreased exercise capacity in hypoxic healthy subjects. METHODS: An incremental cycle ergometer cardiopulmonary exercise test and echocardiographic estimation of pulmonary artery pressure (Ppa) and cardiac output to calculate total PVR were performed in 11 healthy volunteers in normoxia and after 1 h of hypoxic breathing (12% O(2)). The measurements were performed in a random order at 1-week intervals after the receiving either a placebo or bosentan, following a double-blind randomized crossover design. Bosentan was administered twice a day for 3 days, 62.5 mg on the first day and 125 mg on the next 2 days. RESULTS: Hypoxic breathing decreased the mean (+/- SE) pulse oximetric saturation (Spo(2)) from 99 +/- 1% to 3 +/- 1% and increased the mean PVR from 5.6 +/- 0.3 to 7.2 +/- 0.5 mm Hg/L/min/m(2), together with a decrease in mean maximum O(2) uptake (Vo(2)max) from 47 +/- 2 to 35 +/- 2 mL/kg/min. Bosentan had no effect on normoxic measurements and did not affect hypoxic Spo(2), but decreased PVR to 5.6 +/- 0.3 mm Hg/L/min/m(2) (p < 0.01) and increased Vo(2)max to 39 +/- 2 mL/kg/min (p < 0.01) in hypoxia. Bosentan therapy, on average, restored 30% of the hypoxia-induced decrease in Vo(2)max. Bosentan-induced changes in Ppa and Vo(2)max were correlated (p = 0.01). CONCLUSIONS: We conclude that hypoxic pulmonary hypertension partially limits exercise capacity in healthy subjects, and that bosentan therapy can prevent it. PMID- 19188552 TI - Ultrasound assessment for extravascular lung water in patients undergoing hemodialysis. Time course for resolution. AB - BACKGROUND: Sonographic B-lines, also known as lung comets, have been shown to correlate with the presence of extravascular lung water (EVLW). Absent in normal lungs, these sonographic findings become prominent as interstitia and alveoli fill with fluid. Characterization of the dynamics of B-lines, specifically their rate of disappearance as volume is removed, has not been previously described. In this study, we describe the dynamics of B-line resolution in patients undergoing hemodialysis. METHODS: Patients undergoing hemodialysis underwent three chest ultrasound examinations: before, at the midpoint, and after dialysis. We followed a previously described chest ultrasound protocol that counts the number of B lines visualized in 28 lung zones. Baseline demographics, assessment of ejection fraction, time elapsed, net volume of fluid removed, and subjective degree of shortness of breath were recorded for each patient. RESULTS: Forty of 45 patients completed full dialysis runs and had all three lung scans performed; 6 of 40 patients had zero or one B-line predialysis, and none of these 6 patients gained B-lines during dialysis. Thirty-four of 40 patients had statistically significant reductions in the number of B-lines from predialysis to the midpoint scan and from predialysis to postdialysis with a p value < 0.001. There was no association between subjective dyspnea scores and number of B-lines removed. CONCLUSIONS: B line resolution appears to occur real-time as fluid is removed from the body, and this change was statistically significant. These data support thoracic ultrasound as a useful method for evaluating real-time changes in EVLW and in assessing a patient's physiologic response to the removal of fluid. TRIAL REGISTRATION: Massachusetts General Hospital trial registration protocol No. 2007P 002226. PMID- 19188551 TI - Association of the metabolic syndrome with pulmonary venous hypertension. AB - BACKGROUND: Pulmonary venous hypertension (PVH) is a well-described cause of pulmonary hypertension (PH) in patients with left heart disease associated with elevated left heart filling pressure. PVH results from a number of processes, including left-sided valvular disease, constrictive pericardial disease, restrictive cardiomyopathies, and left ventricular (LV) systolic dysfunction. PVH in patients with normal LV systolic function, commonly referred to as diastolic dysfunction, is not well characterized. We observed that many patients with PH due to PVH have obesity, hypertension, diabetes mellitus, and hypercholesterolemia, which are clinical features of the metabolic syndrome (MS), a previously identified cause for systemic vascular disease. METHODS: We evaluated 122 consecutive patients referred for diagnosis and treatment of PH and compared the prevalence of features of the MS between patients with PVH and those with pulmonary arterial hypertension (PAH). We also compared clinical and hemodynamic characteristics between these two groups. RESULTS: Compared to patients with PAH, patients with PVH had a higher frequency of hypertension, obesity, diabetes mellitus, and hyperlipidemia. Two or more features of the MS were found in 16 of 17 patients with PVH (94.1%) compared with 34.3% of patients with PAH (p < 0.001; odds ratio, 30.7; 95% confidence interval, 3.6 to 260.0). PH was substantial, but less severe overall, in patients with PVH compared to those with PAH (mean pulmonary artery pressure, 45 +/- 17 mm Hg [range, 26 to 71 mm Hg] vs 53 +/- 10 [range, 33 to 72 mm Hg], respectively [p = 0.041]; and pulmonary vascular resistance, 4.4 +/- 2.9 units [range, 1.2 to 10.8 units] vs 10.8 +/- 4.7 units [range, 4.8 to 21.9 units], respectively [p < 0.001]). CONCLUSION: PVH is highly associated with the MS. Our results suggest that the MS may predispose patients to develop pulmonary vascular disease. PMID- 19188553 TI - Race/Ethnicity differences in the inpatient management of acute asthma in the United States. AB - BACKGROUND: The only published US study on racial/ethnic disparities in inpatient asthma management was performed in 1989-1990 at a single center. The authors reported that black and Hispanic children were provided substandard care at hospital discharge compared to white children. The purpose of the present analysis was to reexamine this important issue in a large multicenter study. METHODS: A cohort study of 1,232 children and adults hospitalized with a physician diagnosis of acute asthma at 30 hospitals in 22 US states was used. RESULTS: The cohort included 562 children (age range, 2 to 17 years; 39% white; 42% black; and 19% Hispanic) and 670 adults (age range, 18 to 54 years; 44% white; 44% black; and 12% Hispanic). There were no significant racial/ethnic differences in the choice of inpatient medications or length of stay among either children or adults. At hospital discharge, Hispanic children were less likely to receive an asthma action plan (37%) compared to white children (60%) or black children (63%; p < 0.001). Multivariate adjustment for eight variables (including socioeconomic status, hospital admissions for asthma in the past year, medication use prior to presentation, physical examination findings, and hospital admission location) attenuated the statistical significance of this association (odds ratio, 0.5; 95% confidence interval, 0.1 to 2.5). CONCLUSIONS: We did not identify racial/ethnic disparities in the inpatient treatment and outcomes for children or adults with acute asthma. At hospital discharge, Hispanic children were less likely to receive an asthma action plan compared to white or black children, possibly due to language or socioeconomic differences. PMID- 19188554 TI - Cigarette smoking induces overexpression of a fat-depleting gene AZGP1 in the human. AB - BACKGROUND: Smokers weigh less and have less body fat than nonsmokers. Increased body fat and weight gain are observed following smoking cessation. To assess a possible molecular mechanism underlying the inverse association between smoking and body weight, we hypothesized that smoking may induce the expression of a fat depleting gene in the airway epithelium, the cell population that takes the brunt of the stress of cigarette smoke. METHODS: To assess whether smoking up-regulates expression in the airway epithelium of genes associated with weight loss, microarray analysis was used to evaluate genes associated with fat depletion in large airway epithelial samples obtained by fiberoptic bronchoscopy from healthy smokers and healthy nonsmokers. As a candidate gene we further evaluated the expression of alpha(2)-zinc-glycoprotein 1 (AZGP1), a soluble protein that stimulates lipolysis, induces a reduction in body fat in mice, is associated with the cachexia related to cancer, and is known to be expressed in secretory cells of lung epithelium. AZGP1 protein expression was assessed by Western analysis and localization in the large airway epithelium by immunohistochemistry. RESULTS: Both microarray and TaqMan analysis demonstrated that AZGP1 messenger RNA levels were higher in the large airway epithelium of healthy smokers compared to healthy nonsmokers (p < 0.05, all comparisons). Western analysis of airway biopsy specimens from smokers compared with those from nonsmokers demonstrated up regulation of AZGP1 at the protein level, and immunohistochemical analysis demonstrated up-regulation of AZGP1 in secretory as well as neuroendocrine cells of smokers. CONCLUSIONS: In the context that AZGP1 is involved in lipolysis and fat loss, its overexpression in the airway epithelium of chronic smokers may represent one mechanism for the weight difference in smokers vs nonsmokers. PMID- 19188555 TI - Peroxynitrite elevation in exhaled breath condensate of COPD and its inhibition by fudosteine. AB - BACKGROUND: Peroxynitrite (PN) formed by the reaction of nitric oxide and superoxide is a powerful oxidant/nitrosant. Nitrative stress is implicated in COPD pathogenesis, but PN has not been detected due to a short half-life (< 1 s) at physiologic condition. Instead, 3-nitrotyrosine has been measured as a footprint of PN release. METHOD: PN was measured using oxidation of 2',7' dichlorofluorescein (DCDHF) in exhaled breath condensate (EBC) collected in high pH and sputum cells. The PN scavenging effect was also evaluated by the same system as PN-induced bovine serum albumin (BSA) nitration. RESULTS: The mean (+/- SD) PN levels in EBC of COPD patients (7.9 +/- 3.0 nmol/L; n = 10) were significantly higher than those of healthy volunteers (2.0 +/- 1.1 nmol/L; p < 0.0001; n = 8) and smokers (2.8 +/- 0.9 nmol/L; p = 0.0017; n = 6). There was a good correlation between PN level and disease severity (FEV(1)) in COPD (p = 0.0016). Fudosteine (FDS), a unique mucolytic antioxidant, showed a stronger scavenging effect of PN than N-acetyl-cysteine on DCDHF oxidation in vitro and in sputum macrophages, and also on PN-induced BSA nitration. FDS (0.1 mmol/L) reduced PN-enhanced interleukin (IL)-1beta-induced IL-8 release and restored corticosteroid sensitivity defected by PN more potently than those induced by H(2)O(2) in A549 airway epithelial cells. CONCLUSION: This noninvasive PN measurement in EBC may be useful for monitoring airway nitrative stress in COPD. Furthermore, FDS has the potential to inhibit PN-induced events in lung by its scavenging effect. PMID- 19188556 TI - Clara cell protein (CC16), a marker of lung epithelial injury, is decreased in plasma and pulmonary edema fluid from patients with acute lung injury. AB - BACKGROUND: Acute lung injury (ALI) and ARDS are common clinical syndromes that are underdiagnosed. Clara cell secretory protein (CC16) is an antiinflammatory protein secreted by the Clara cells of the distal respiratory epithelium that has been proposed as a biomarker of lung epithelial injury. We tested the diagnostic and prognostic utility of CC16 in patients with non-trauma-related ALI/ARDS compared to a control group of patients with acute cardiogenic pulmonary edema (CPE). METHODS: Plasma and pulmonary edema fluid samples were obtained from medical and surgical patients with ALI/ARDS or CPE requiring intubation for mechanical ventilation. The etiology of pulmonary edema was determined using consensus clinical criteria for ALI/ARDS and CPE and the edema fluid-to-plasma protein ratio. Plasma and edema fluid CC16 levels were measured by sandwich enzyme-linked immunosorbent assay. CC16 levels were log transformed for analysis, and comparisons were made by the Student t test or Chi(2) as appropriate. RESULTS: Compared to patients with CPE (n = 9), patients with ALI/ARDS (n = 23) had lower median CC16 levels in plasma (22 ng/mL [interquartile range (IQR), 9 to 44 ng/mL] vs 55 ng/mL [IQR, 18 to 123 ng/mL], respectively; p = 0.053) and pulmonary edema fluid (1,950 ng/mL [IQR, 1,780 to 4,024 ng/mL] vs 4,835 ng/mL [IQR, 2,006 to 6,350 ng/mL], respectively; p = 0.044). Relative to total pulmonary edema fluid protein concentration, the median CC16 level was significantly lower in patients with ALI/ARDS (45 ng CC16/mg total protein [IQR, 4 to 64 ng CC16/mg total protein] vs 120 ng CC16/mg total protein [IQR, 87 to 257 ng CC16/mg total protein], respectively; p = 0.005). Neither plasma nor edema fluid CC16 levels predicted mortality, the number of days of unassisted ventilation, or ICU length of stay. CONCLUSION: CC16 is a promising diagnostic biomarker for helping to discriminate ALI from CPE. Larger scale validation is warranted to better characterize the utility of CC16 in the diagnosis of this underrecognized syndrome. PMID- 19188557 TI - Specific IgE response to trichophyton and asthma severity. AB - BACKGROUND: Sensitization to Trichophyton, a major dermatophyte, has been associated with asthma. Whether such sensitization is generally associated with the severity of asthma, like other molds such as Alternaria, is unknown. METHODS: We compared 258 patients with asthma, which was classified by severity as mild (n = 123), moderate (101), or severe (34), and 114 healthy control subjects, with regard to specific IgE titers against Trichophyton rubrum and other common allergens such as mixed molds, house-dust mite, cat dander, dog dander, Japanese cedar pollen, mixed Graminea pollens and mixed weed pollens. RESULTS: Positive rate of Trichophyton-specific IgE was higher in the patients with moderate asthma (15.8%) than in the control subjects (7.0%, p = 0.04) and patients with mild asthma (4.9%, p < 0.006), and it was also higher in the patients with severe asthma (32.4%) than in control subjects (p = 0.0001), and patients with mild asthma (p < 0.0001) and moderate asthma (p = 0.04), but it did not differ between the control subjects and patients with mild asthma. The positive rates of mixed molds, cat dander, and dog dander were almost invariably higher in patients in all asthma subgroups than in the control subjects but did not differ among patients in the three asthma subgroups. The positive rates of other allergens were not different in all groups. Reanalysis of positive rate of Trichophyton specific IgE after excluding 52 subjects with positive results for mixed molds showed a similar statistical trend to that of the original cohort. This may negate the potential effect of cross-reactivity to these molds. Multivariate analysis of asthma subgroups identified positive IgE results for Trichophyton as an independent determinant of asthma severity. CONCLUSIONS: Specific IgE response to Trichophyton may be associated with more severe asthma. PMID- 19188558 TI - Comparative study of navigated versus freehand osteochondral graft transplantation of the knee. AB - BACKGROUND: Osteochondral lesions are a common sports-related injury for which osteochondral grafting, including mosaicplasty, is an established treatment. Computer navigation has been gaining popularity in orthopaedic surgery to improve accuracy and precision. HYPOTHESIS: Navigation improves angle and depth matching during harvest and placement of osteochondral grafts compared with conventional freehand open technique. STUDY DESIGN: Controlled laboratory study. METHODS: Three cadaveric knees were used. Reference markers were attached to the femur, tibia, and donor/recipient site guides. Fifteen osteochondral grafts were harvested and inserted into recipient sites with computer navigation, and 15 similar grafts were inserted freehand. The angles of graft removal and placement as well as surface congruity (graft depth) were calculated for each surgical group. RESULTS: The mean harvesting angle at the donor site using navigation was 4 degrees (standard deviation, 2.3 degrees ; range, 1 degrees -9 degrees ) versus 12 degrees (standard deviation, 5.5 degrees ; range, 5 degrees -24 degrees ) using freehand technique (P < .0001). The recipient plug removal angle using the navigated technique was 3.3 degrees (standard deviation, 2.1 degrees ; range, 0 degrees -9 degrees ) versus 10.7 degrees (standard deviation, 4.9 degrees ; range, 2 degrees -17 degrees ) in freehand (P < .0001). The mean navigated recipient plug placement angle was 3.6 degrees (standard deviation, 2.0 degrees ; range, 1 degrees -9 degrees ) versus 10.6 degrees (standard deviation, 4.4 degrees ; range, 3 degrees -17 degrees ) with freehand technique (P = .0001). The mean height of plug protrusion under navigation was 0.3 mm (standard deviation, 0.2 mm; range, 0-0.6 mm) versus 0.5 mm (standard deviation, 0.3 mm; range, 0.2 1.1 mm) using a freehand technique (P = .0034). CONCLUSION: Significantly greater accuracy and precision were observed in harvesting and placement of the osteochondral grafts in the navigated procedures. Clinical studies are needed to establish a benefit in vivo. CLINICAL RELEVANCE: Improvement in the osteochondral harvest and placement is desirable to optimize clinical outcomes. Navigation shows great potential to improve both harvest and placement precision and accuracy, thus optimizing ultimate surface congruity. PMID- 19188559 TI - Biomechanics of supination ankle sprain: a case report of an accidental injury event in the laboratory. PMID- 19188560 TI - Management of tendinopathy. AB - Overuse disorders of tendons, or tendinopathies, present a challenge to sports physicians, surgeons, and other health care professionals dealing with athletes. The Achilles, patellar, and supraspinatus tendons are particularly vulnerable to injury and often difficult to manage successfully. Inflammation was believed central to the pathologic process, but histopathologic evidence has confirmed the failed healing response nature of these conditions. Excessive or inappropriate loading of the musculotendinous unit is believed to be central to the disease process, although the exact mechanism by which this occurs remains uncertain. Additionally, the location of the lesion (for example, the midtendon or osteotendinous junction) has become increasingly recognized as influencing both the pathologic process and subsequent management. The mechanical, vascular, neural, and other theories that seek to explain the pathologic process are explored in this article. Recent developments in the nonoperative management of chronic tendon disorders are reviewed, as is the rationale for surgical intervention. Recent surgical advances, including minimally invasive tendon surgery, are reviewed. Potential future management strategies, such as stem cell therapy, growth factor treatment, and gene transfer, are also discussed. PMID- 19188561 TI - A prospective comparison of 3 hamstring ACL fixation devices--Rigidfix, BioScrew, and Intrafix--randomized into 4 groups with 2 years of follow-up. AB - BACKGROUND: New devices for graft fixation in anterior cruciate ligament reconstruction are released to clinical use without clinical follow-up data. HYPOTHESIS: There is similar clinical outcome after either cross-pin or absorbable interference screw fixation in anterior cruciate ligament reconstruction with hamstring tendons. STUDY DESIGN: Randomized controlled clinical trial; Level of evidence, 1. METHODS: A total of 120 patients were randomized into 4 different groups (30 each) for anterior cruciate ligament reconstruction with hamstring tendons: femoral Rigidfix cross-pin and Intrafix tibial expansion sheath with a tapered expansion screw; Rigidfix femoral and BioScrew interference screw tibial fixation, BioScrew femoral and Intrafix tibial fixation; or BioScrew fixation into both tunnels. The evaluation methods were clinical examination, knee scores, and instrumented laxity measurements. RESULTS: Ten patients were completely lost to follow-up and 3 revisions were done before the 2-year follow-up, leaving 107 of 120 (89%) patients for analysis. No statistically significant differences between the groups were seen 2 years postoperatively, and all but 2 patients in the Rigidfix/Intrafix and Rigidfix/BioScrew groups, respectively, were classified into International Knee Documentation Committee A or B categories. A revision reconstruction was performed before the 2-year follow-up in 2 cases after a high-energy injury caused a rerupture (1 in Rigidfix/Intrafix and 1 in BioScrew/BioScrew groups). In addition, there were 4 nontraumatic failures revised before the 2-year follow-up (2 in Rigidfix/Intrafix and 1 each in Rigidfix/BioScrew and BioScrew/BioScrew). CONCLUSION: There were no statistically or clinically relevant differences in the results 2 years postoperatively, and all 4 techniques improved patient performance. PMID- 19188562 TI - Arthroscopic versus open acromioplasty: a meta-analysis. AB - BACKGROUND: To address persisting controversy in the literature concerning the efficacy of arthroscopic compared to open acromioplasty, a meta-analysis was performed to evaluate the treatment effect after both approaches. HYPOTHESIS: The final clinical outcomes will be the same after both open and arthroscopic acromioplasty. However, the arthroscopic technique results in faster recovery and less postoperative morbidity as reflected by faster return to work and decreased hospital stays. STUDY DESIGN: Meta-analysis; Level of evidence, 3. METHODS: We performed our search of published English language literature using PubMed. We also searched the proceedings from 4 major orthopaedic meetings convened from 2000 to 2007. Furthermore, the reference sections of all relevant articles were reviewed for pertinent studies and presentations. Nine studies met the inclusion criteria that directly compared arthroscopic versus open acromioplasty with minimum follow-up of 1 year. The analysis focused on 1-year clinical outcome and included comparison of the objective 100-point score, hospital stay, time until return to work, operative time, and complications. RESULTS: No significant differences were found in clinical outcomes or complications for the 2 groups. However, open acromioplasty was associated with longer hospital stays (2.3 days, P = .05) and a greater length in time until return to work (65.1 days) compared with the arthroscopic technique (48.6 days) (P < .05). CONCLUSION: Arthroscopic and open acromioplasty have equivalent ultimate clinical outcomes, operative times, and low complication rates. However, arthroscopic acromioplasty results in faster return to work and fewer hospital inpatient days compared with the open technique. PMID- 19188563 TI - Dynamic optimization of chronic migraine treatment: current and future options. AB - Comparator studies that assess treatment effects in a clinical setting have improved the understanding of the efficacy and tolerability of prophylactic treatments for chronic migraine (CM). It is premature to recommend device-based treatments, such as occipital nerve stimulation, vagal nerve stimulation, and patent foramen ovale closure for CM, because clinical trials are in the preliminary stages. Physical therapy techniques, like applying heat or cold packs, ultrasonography, and electrical stimulation, have been shown to lessen pain. Nonpharmacologic treatments, including cognitive behavioral therapy, stress management, and biofeedback, have been investigated and proved effective in some areas of pain management, including migraine. However, pharmacologic interventions may be necessary for effective, long-term prophylaxis. Several medications under investigation, including topiramate, gabapentin, tizanidine, and amitriptyline, have proved efficacious in reducing the number of migraine episodes and the pain associated with migraine, although adverse events may prevent continued use of some agents. Evidence supports the use of botulinum toxin type A (BoNT-A) for CM, with or without medication overuse, to achieve a significant reduction in headache episodes. Efficacy of BoNT-A for CM is comparable with or better than that of valproate and topiramate, with better tolerability. Predictors of response to BoNT-A for CM appear to include predominantly unilateral location of the headache and the presence of cutaneous or muscle allodynia. BoNT-A has been demonstrated to be safe and well tolerated, with rare discontinuations due to adverse events. Recent clinical trials indicate that rational combination therapy may have a place in treating refractory CM. Well-controlled multicenter trials are awaited. PMID- 19188564 TI - Tracing transformation: chronic migraine classification, progression, and epidemiology. AB - Migraine attacks sometimes increase in frequency over time. Headache experts conceptualize this process with a model that envisions transition into and out of four distinct states: no migraine, low-frequency episodic migraine (<10 headaches per month), high-frequency episodic migraine (10-14 headaches per month), and chronic migraine (CM, >or=15 headaches per month). Transitions may be in the direction of increasing or decreasing headache frequency and are influenced by specific risk factors. Overall, population studies estimate that patients who have low-frequency episodic migraine or high-frequency episodic migraine will transition to CM at the rate of about 2.5% per year. Two longitudinal population studies, the Frequent Headache Epidemiology study and the ongoing American Migraine Prevalence and Prevention (AMPP) study provide longitudinal population data that has defined the rates of and risk factors for transition. Launched in 2004, the AMPP study has followed a sample of >10,000 migraine sufferers annually for 4 years. Cross-sectional data from the Frequent Headache Epidemiology study and the AMPP study show that patients with chronic daily headaches have lower levels of education and household income. In addition, epidemiologic profiles show that CM sufferers tend to be older and have higher body mass indexes. These studies have also assessed a number of potential risk factors associated with the transition to CM. These include baseline high attack frequency, obesity, stressful life events, snoring, and overuse of certain classes of medication. In particular, opiate and barbiturate combination products contribute to migraine progression, and nonsteroidal anti-inflammatory agents are protective in patients with <10 headache days per month. The influence of medication is modified by both headache attack frequency and frequency of medication use. Although depression and anxiety are associated with an increased risk of new-onset CM, the influence of depression is accounted for by migraine disability assessment scale score, whereas the effect of anxiety may be independent of migraine disability assessment scale score. Emerging data on the longitudinal risk of CM suggest that, in a population at risk, CM may be a preventable disorder. PMID- 19188565 TI - Spectrum of illness: understanding biological patterns and relationships in chronic migraine. AB - Chronic migraine (CM) typically evolves from episodic migraine (EM) over months to years in susceptible individuals. Headaches increase in frequency over time, becoming less intense but more disabling and less responsive to treatment. The results of electrophysiologic and functional imaging studies indicate that CM is associated with abnormalities in the periaqueductal gray matter that may be progressive. In addition, CM is associated with a greater degree of impairment in cortical processing of sensory stimuli than EM, perhaps because of more pervasive or persistent cortical hyperexcitability. These findings fit with the model of migraine as a spectrum disorder, in which the clinical and pathophysiologic features of migraine may progress over time. This progression is postulated to result from changes in nociceptive thresholds and ensuing central sensitization caused by recurrent migraine in susceptible individuals, for whom risk factors have been described. Also, progression may lead to changes in baseline neurologic function between episodes of headache, evident in electrophysiologic and functional imaging studies and as an increase in depression, anxiety, nonhead pain, fatigue, gastrointestinal disorders, and other somatic complaints that may occur after years of EM. From the available research and migraine models, a concept of CM is emerging that identifies relatively permanent and pervasive central changes warranting novel, tolerable treatments. This model also implies that prevention of CM is an important goal in the management of EM, particularly for individuals who exhibit risk factors for chronic transformation. PMID- 19188566 TI - International issues: of saints and sickness: a neurology elective in India. PMID- 19188567 TI - Teaching NeuroImage: idiopathic hypertrophic spinal pachymeningitis. PMID- 19188569 TI - Natalizumab: bound to rebound? PMID- 19188570 TI - Cortical malformations: looking behind the cortex. PMID- 19188571 TI - Effect of plasma exchange in accelerating natalizumab clearance and restoring leukocyte function. AB - BACKGROUND: Accelerating the clearance of therapeutic monoclonal antibodies (mAbs) from the body may be useful to address uncommon but serious complications from treatment, such as progressive multifocal leukoencephalopathy (PML). Treatment of PML requires immune reconstitution. Plasma exchange (PLEX) may accelerate mAb clearance, restoring the function of inhibited proteins and increasing the number or function of leukocytes entering the CNS. We evaluated the efficacy of PLEX in accelerating natalizumab (a therapy for multiple sclerosis [MS] and Crohn disease) clearance and alpha4-integrin desaturation. Restoration of leukocyte transmigratory capacity was evaluated using an in vitro blood-brain barrier (ivBBB). METHODS: Twelve patients with MS receiving natalizumab underwent three 1.5-volume PLEX sessions over 5 or 8 days. Natalizumab concentrations and alpha4-integrin saturation were assessed daily throughout PLEX and three times over the subsequent 2 weeks, comparing results with the same patients the previous month. Peripheral blood mononuclear cell (PBMC) migration (induced by the chemokine CCL2) across an ivBBB was assessed in a subset of six patients with and without PLEX. RESULTS: Serum natalizumab concentrations were reduced by a mean of 92% from baseline to 1 week after three PLEX sessions (p < 0.001). Although average alpha4-integrin saturation was not reduced after PLEX, it was reduced to less than 50% when natalizumab concentrations were below 1 mug/mL. PBMC transmigratory capacity increased 2.2 fold after PLEX (p < 0.006). CONCLUSIONS: Plasma exchange (PLEX) accelerated clearance of natalizumab, and at natalizumab concentrations below 1 mug/mL, desaturation of alpha4-integrin was observed. Also, CCL2-induced leukocyte transmigration across an in vitro blood-brain barrier was increased after PLEX. Therefore, PLEX may be effective in restoring immune effector function in natalizumab-treated patients. PMID- 19188572 TI - Assessment of potential drug interactions in patients with epilepsy: impact of age and sex. AB - OBJECTIVES: To understand and quantify the exposure to concomitant medications other than antiepileptic drugs (AEDs) within an age-diverse group of men and women with epilepsy and explore the likelihood of relevant drug interactions as a result. METHODS: The PharMetrics medical and pharmaceutical claims database was used to extract data for commercially insured adult patients with a diagnosis of epilepsy and treated with any AED during the period from July 1, 2001, to December 31, 2004. Data were analyzed for concomitant non-AEDs used after initiating AEDs in six age groups, spanning the ages 18 to 85+ years, in both men and women. RESULTS: Use of concomitant medications occurred in every age group and increased with age for both men and women (mean number of non-AEDs ranging from 2.41 to 7.67 in males aged 18-34 and 85+ years and from 4.04 to 7.05 in females aged 18-34 and 85+ years; p < 0.001 for age trend). beta-Hydroxy-beta methylglutaryl-coenzyme A reductase inhibitors (statins), calcium channel blockers (CCBs), and selective serotonin reuptake inhibitors (SSRIs) were the most commonly used non-AED medications with the potential for adverse drug interactions. SSRIs use was substantial in all age groups and greater than for statins or CCBs in patients aged 18-54 years. Use of antipsychotics, tricyclic antidepressants, and warfarin was also noted in more than 10% of patients across different age groups. CONCLUSIONS: Polypharmacy with non-antiepileptic drug (AED) medications is common in both men and women, and is not a situation unique to only elderly patients with epilepsy. In particular, use of potentially interacting, enzyme inducing AEDs was common. These findings suggest that clinicians must be mindful of potential AED-non-AED drug interactions, in patients of all age groups. PMID- 19188573 TI - Automatic detection of preclinical neurodegeneration: presymptomatic Huntington disease. AB - BACKGROUND: Treatment of neurodegenerative diseases is likely to be most beneficial in the very early, possibly preclinical stages of degeneration. We explored the usefulness of fully automatic structural MRI classification methods for detecting subtle degenerative change. The availability of a definitive genetic test for Huntington disease (HD) provides an excellent metric for judging the performance of such methods in gene mutation carriers who are free of symptoms. METHODS: Using the gray matter segment of MRI scans, this study explored the usefulness of a multivariate support vector machine to automatically identify presymptomatic HD gene mutation carriers (PSCs) in the absence of any a priori information. A multicenter data set of 96 PSCs and 95 age- and sex-matched controls was studied. The PSC group was subclassified into three groups based on time from predicted clinical onset, an estimate that is a function of DNA mutation size and age. RESULTS: Subjects with at least a 33% chance of developing unequivocal signs of HD in 5 years were correctly assigned to the PSC group 69% of the time. Accuracy improved to 83% when regions affected by the disease were selected a priori for analysis. Performance was at chance when the probability of developing symptoms in 5 years was less than 10%. CONCLUSIONS: Presymptomatic Huntington disease gene mutation carriers close to estimated diagnostic onset were successfully separated from controls on the basis of single anatomic scans, without additional a priori information. Prior information is required to allow separation when degenerative changes are either subtle or variable. PMID- 19188574 TI - Incidence and remaining lifetime risk of Parkinson disease in advanced age. AB - OBJECTIVE: To estimate the incidence and lifetime risk (LTR) of Parkinson disease (PD) in a large cohort of men. BACKGROUND: Age is the strongest risk factor for PD, but whether its incidence continues to increase after age 80 years remains unclear. METHODS: Prospective cohort of 21,970 US male physicians aged 40-84 years at baseline who did not report PD before study entry. Participants self reported PD on yearly follow-up questionnaires, and all deaths were confirmed. We calculated incidence rates and cumulative incidence using a modified Kaplan-Meier analysis. LTR was estimated by adjusting cumulative incidence for competing risks of death. RESULTS: Five hundred sixty-three cases of PD were identified over 23 years of follow-up. The crude incidence rate of PD was 121 cases/100,000 person years. Age-specific incidence rates increased sharply beginning at age 60 years, peaked in those aged 85-89 years, and declined beginning at age 90 years. Cumulative incidence substantially overestimated the long-term risk of PD, particularly in those aged 80 years and older. Cumulative incidence was 9.9% (95% confidence interval [CI] 8.48%-11.30%) from ages 45 to 100 years, whereas LTR for the same period was 6.7% (95% CI 6.01%-7.43%). The incidence and LTR of PD decreased with increasing exposure to smoking. CONCLUSIONS: Our study provides evidence that the incidence of Parkinson disease (PD) in men increases through age 89 years. Whether the subsequent decline represents a true decrease in risk remains to be established. A history of smoking substantially decreased the incidence and lifetime risk of PD. Incidence studies that do not adjust for competing risks of death may overestimate the true risk of PD in the elderly. PMID- 19188576 TI - Clinical and electrodiagnostic correlates of peroneal intraneural ganglia. AB - OBJECTIVE: Intraneural ganglia (IG) are an underappreciated but treatable cause of common peroneal neuropathy (CPN). This study was designed to determine if there are clinical measures that distinguish CPN caused by IG from CPN without a clear proximate cause. METHODS: Clinical and electrodiagnostic features of 22 cases of IG were compared in a case-control study to 11 cases of CPN with imaging negative for IG. RESULTS: The IG group had a greater body mass index (30 vs 24; p < 0.005), more pain at the knee (52% of 22 vs 0% of 11; p < 0.005) or in the peroneal distribution (76% of 21 vs 27% of 1; p < 0.02), more frequent fluctuating weakness (48% of 21 vs 4% of 29; p < 0.01) with weight bearing (38%, p < 0.05), or a palpable mass (47% of 20, p < 0.01) at the fibular head. The IG group was less likely to present with a history of weight loss (0% vs 36%; p < 0.01), immobility (0% vs 21%; p < 0.03), or leg crossing (0% vs 80%; p < 0.05). There were no significant electrophysiologic differences. CONCLUSIONS: Presenting clinical features increase the likelihood of intraneural ganglia and may assist selection of patients with common peroneal neuropathy for diagnostic peroneal nerve imaging. PMID- 19188575 TI - Randomized, double-blind, placebo-controlled study of XP13512/GSK1838262 in patients with RLS. AB - OBJECTIVE: To assess the efficacy and tolerability of the nondopaminergic agent XP13512/GSK1838262 in adults with moderate to severe primary restless legs syndrome (RLS). METHODS: Patient Improvements in Vital Outcomes following Treatment in Restless Legs Syndrome I was a 12-week, multicenter, randomized, double-blind, placebo-controlled trial of XP13512 1,200 mg or placebo taken once daily at 5:00 pm with food. Coprimary endpoints were mean change from baseline International Restless Legs Scale (IRLS) total score and proportion of investigator-rated responders (very much improved or much improved on the Clinical Global Impression-Improvement scale) at week 12 (last observation carried forward). Tolerability was assessed using adverse events, vital signs, and clinical laboratory parameters. RESULTS: A total of 222 patients were randomized (XP13512 = 114, placebo = 108) and 192 patients (XP13512 = 100, placebo = 92) completed the study. At week 12, the mean change from baseline IRLS total score was greater with XP13512 (-13.2) compared with placebo (-8.8). Analysis of covariance, adjusted for baseline score and pooled site, demonstrated a mean treatment difference of -4.0 (95% confidence interval [CI], -6.2 to -1.9; p = 0.0003). More patients treated with XP13512 (76.1%) were responders compared with placebo (38.9%; adjusted OR 5.1; 95% CI, 2.8 to 9.2; p < 0.0001). Significant treatment effects for both coprimary measures were identified at week 1, the earliest time point measured. The most commonly reported adverse events were somnolence (XP13512 27%, placebo 7%) and dizziness (XP13512 20%, placebo 5%), which were mild to moderate in intensity and generally remitted. CONCLUSIONS: XP13512 1,200 mg, taken once daily, significantly improved restless legs syndrome (RLS) symptoms compared with placebo and was generally well tolerated in adults with moderate to severe primary RLS. PMID- 19188577 TI - Executive dysfunction in frontotemporal dementia and corticobasal syndrome. AB - OBJECTIVE: To determine the pattern of executive dysfunction in frontotemporal dementia (FTD) and corticobasal syndrome (CBS) and to determine the brain areas associated with executive dysfunction in these illnesses. METHOD: We administered the Delis-Kaplan Executive Function System (D-KEFS), a collection of standardized executive function tests, to 51 patients with behavioral-variant FTD and 50 patients with CBS. We also performed a discriminant analysis on the D-KEFS to determine which executive function tests best distinguished the clinical diagnoses of FTD and CBS. Finally, we used voxel-based morphometry (VBM) to determine regional gray matter volume loss associated with executive dysfunction. RESULTS: Patients with FTD and patients with CBS showed executive dysfunction greater than memory dysfunction. Executive function was better preserved in the patients with CBS than the patients with FTD with the exception of tests that required motor, visuospatial ability, or both. In patients with CBS, dorsal frontal and parietal and temporal-parietal cortex was associated with executive function. In FTD, tests with a language component (Verbal Fluency) were associated with left perisylvian cortex, sorting with the left dorsolateral prefrontal cortex, and reasoning (the Twenty Questions task) with the left anterior frontal cortex. The Twenty Questions test best distinguished the clinical diagnoses of CBS and FTD. CONCLUSIONS: The neuroanatomic findings (especially in frontotemporal dementia [FTD]) agree with the previous literature on this topic. Patients with FTD and patients with corticobasal syndrome (CBS) show disparate performance on higher-order executive functions, especially the Twenty Questions test. It may be difficult to distinguish motor and visuospatial ability from executive function in patients with CBS using tests with significant motor and visuospatial demands such as Trail Making. PMID- 19188579 TI - Ocular motor and lid apraxia as initial symptom of anti-Ma1/Ma2-associated encephalitis. PMID- 19188580 TI - Atypical voluntary nystagmus. PMID- 19188578 TI - Educational attainment and cognitive decline in old age. AB - BACKGROUND: Level of education is a well-established risk factor for Alzheimer disease but its relation to cognitive decline, the principal clinical manifestation of the disease, is uncertain. METHODS: More than 6,000 older residents of a community on the south side of Chicago were interviewed at approximately 3-year intervals for up to 14 years. The interview included administration of four brief tests of cognitive function from which a previously established composite measure of global cognition was derived. We estimated the associations of education with baseline level of cognition and rate of cognitive change in a series of mixed-effects models. RESULTS: In an initial analysis, higher level of education was related to higher level of cognition at baseline, but there was no linear association between education and rate of change in cognitive function. In a subsequent analysis with terms to allow for nonlinearity in education and its relation to cognitive decline, rate of cognitive decline at average or high levels of education was slightly increased during earlier years of follow-up but slightly decreased in later years in comparison to low levels of education. Findings were similar among black and white participants. Cognitive performance improved with repeated test administration, but there was no evidence that retest effects were related to education or attenuated education's association with cognitive change. CONCLUSIONS: The results suggest that education is robustly associated with level of cognitive function but not with rate of cognitive decline and that the former association primarily accounts for education's correlation with risk of dementia in old age. PMID- 19188581 TI - Direct visualization of remyelination in multiple sclerosis using T2-weighted high-field MRI. PMID- 19188582 TI - The ALS/PDC syndrome of Guam and the cycad hypothesis. PMID- 19188583 TI - Is it time for neurohospitalists? PMID- 19188584 TI - American fast food isn't all corn-based. PMID- 19188585 TI - Thymic stromal lymphopoietin overproduced by keratinocytes in mouse skin aggravates experimental asthma. AB - Atopic dermatitis (AD) is often the initial step in the "atopic march," given that more than half of AD patients with moderate to severe AD develop asthma later in life. Both AD and asthma share a similar "atopy" phenotype that includes T helper type 2 inflammation with eosinophilia and hyper-IgE immunoglobulinemia, but the molecular mechanisms underlying the "atopic march" remain elusive. In the present study, we show that induced expression of thymic stromal lymphopoietin (TSLP) in mouse epidermal keratinocytes upon topical application of MC903 (a low calcemic analogue of vitamin D3) not only triggers AD as we previously reported but also aggravates experimental allergic asthma induced by ovalbumin sensitization and challenge. Our study, which provides a mouse model to study human "atopic march," indicates that keratinocyte-produced TSLP may represent an important factor in the link of atopic dermatitis to asthma. PMID- 19188587 TI - Climate change and health costs of air emissions from biofuels and gasoline. AB - Environmental impacts of energy use can impose large costs on society. We quantify and monetize the life-cycle climate-change and health effects of greenhouse gas (GHG) and fine particulate matter (PM(2.5)) emissions from gasoline, corn ethanol, and cellulosic ethanol. For each billion ethanol equivalent gallons of fuel produced and combusted in the US, the combined climate change and health costs are $469 million for gasoline, $472-952 million for corn ethanol depending on biorefinery heat source (natural gas, corn stover, or coal) and technology, but only $123-208 million for cellulosic ethanol depending on feedstock (prairie biomass, Miscanthus, corn stover, or switchgrass). Moreover, a geographically explicit life-cycle analysis that tracks PM(2.5) emissions and exposure relative to U.S. population shows regional shifts in health costs dependent on fuel production systems. Because cellulosic ethanol can offer health benefits from PM(2.5) reduction that are of comparable importance to its climate change benefits from GHG reduction, a shift from gasoline to cellulosic ethanol has greater advantages than previously recognized. These advantages are critically dependent on the source of land used to produce biomass for biofuels, on the magnitude of any indirect land use that may result, and on other as yet unmeasured environmental impacts of biofuels. PMID- 19188586 TI - Loss of cryptochrome reduces cancer risk in p53 mutant mice. AB - It is commonly thought that disruption of the circadian clock increases the cancer incidence in humans and mice. However, it was found that disruption of the clock by the Cryptochrome (Cry) mutation in mice did not increase cancer rate in the mutant mice even after exposing the animals to ionizing radiation. Therefore, in this study we tested the effect of the Cry mutation on carcinogenesis in a mouse strain prone to cancer because of a p53 mutation, with the expectation that clock disruption in this sensitized background would further increase cancer risk. Paradoxically, we find that the Cry mutation protects p53 mutant mice from the early onset of cancer and extends their median lifespan approximately 50%, in part by sensitizing p53 mutant cells to apoptosis in response to genotoxic stress. These results suggest alternative therapeutic approaches in management of cancers associated with a p53 mutation. PMID- 19188588 TI - Kernel energy method applied to vesicular stomatitis virus nucleoprotein. AB - The kernel energy method (KEM) is applied to the vesicular stomatitis virus (VSV) nucleoprotein (PDB ID code 2QVJ). The calculations employ atomic coordinates from the crystal structure at 2.8-A resolution, except for the hydrogen atoms, whose positions were modeled by using the computer program HYPERCHEM. The calculated KEM ab initio limited basis Hartree-Fock energy for the full 33,175 atom molecule (including hydrogen atoms) is obtained. In the KEM, a full biological molecule is represented by smaller "kernels" of atoms, greatly simplifying the calculations. Collections of kernels are well suited for parallel computation. VSV consists of five similar chains, and we obtain the energy of each chain. Interchain hydrogen bonds contribute to the interaction energy between the chains. These hydrogen bond energies are calculated in Hartree-Fock (HF) and Moller-Plesset perturbation theory to second order (MP2) approximations by using 6-31G** basis orbitals. The correlation energy, included in MP2, is a significant factor in the interchain hydrogen bond energies. PMID- 19188589 TI - Metamaterials and the Landau-Lifshitz permeability argument: large permittivity begets high-frequency magnetism. AB - Homogeneous composites, or metamaterials, made of dielectric or metallic particles are known to show magnetic properties that contradict arguments by Landau and Lifshitz [Landau LD, Lifshitz EM (1960) Electrodynamics of Continuous Media (Pergamon, Oxford, UK), p 251], indicating that the magnetization and, thus, the permeability, loses its meaning at relatively low frequencies. Here, we show that these arguments do not apply to composites made of substances with Im square root(epsilon(S)) >> lambda/l or Re square root(epsilon(S)) approximately lambda/l (epsilon(S) and l are the complex permittivity and the characteristic length of the particles, and lambda >> l is the vacuum wavelength). Our general analysis is supported by studies of split rings, one of the most common constituents of electromagnetic metamaterials, and spherical inclusions. An analytical solution is given to the problem of scattering by a small and thin split ring of arbitrary permittivity. Results reveal a close relationship between epsilon(S) and the dynamic magnetic properties of metamaterials. For |square root(epsilon(S))| << lambda/a (a is the ring cross-sectional radius), the composites exhibit very weak magnetic activity, consistent with the Landau Lifshitz argument and similar to that of molecular crystals. In contrast, large values of the permittivity lead to strong diamagnetic or paramagnetic behavior characterized by susceptibilities whose magnitude is significantly larger than that of natural substances. We compiled from the literature a list of materials that show high permittivity at wavelengths in the range 0.3-3000 microm. Calculations for a system of spherical inclusions made of these materials, using the magnetic counterpart to Lorentz-Lorenz formula, uncover large magnetic effects the strength of which diminishes with decreasing wavelength. PMID- 19188590 TI - Aberrant miR-182 expression promotes melanoma metastasis by repressing FOXO3 and microphthalmia-associated transcription factor. AB - The highly aggressive character of melanoma makes it an excellent model for probing the mechanisms underlying metastasis, which remains one of the most difficult challenges in treating cancer. We find that miR-182, member of a miRNA cluster in a chromosomal locus (7q31-34) frequently amplified in melanoma, is commonly up-regulated in human melanoma cell lines and tissue samples; this up regulation correlates with gene copy number in a subset of melanoma cell lines. Moreover, miR-182 ectopic expression stimulates migration of melanoma cells in vitro and their metastatic potential in vivo, whereas miR-182 down-regulation impedes invasion and triggers apoptosis. We further show that miR-182 over expression promotes migration and survival by directly repressing microphthalmia associated transcription factor-M and FOXO3, whereas enhanced expression of either microphthalmia-associated transcription factor-M or FOXO3 blocks miR-182's proinvasive effects. In human tissues, expression of miR-182 increases with progression from primary to metastatic melanoma and inversely correlates with FOXO3 and microphthalmia-associated transcription factor levels. Our data provide a mechanism for invasion and survival in melanoma that could prove applicable to metastasis of other cancers and suggest that miRNA silencing may be a worthwhile therapeutic strategy. PMID- 19188591 TI - Disentangling prenatal and inherited influences in humans with an experimental design. AB - Exposure to adversity in utero at a sensitive period of development can bring about physiological, structural, and metabolic changes in the fetus that affect later development and behavior. However, the link between prenatal environment and offspring outcomes could also arise and confound because of the relation between maternal and offspring genomes. As human studies cannot randomly assign offspring to prenatal conditions, it is difficult to test whether in utero events have true causal effects on offspring outcomes. We used an unusual approach to overcome this difficulty whereby pregnant mothers are either biologically unrelated or related to their child as a result of in vitro fertilization (IVF). In this sample, prenatal smoking reduces offspring birth weight in both unrelated and related offspring, consistent with effects arising through prenatal mechanisms independent of the relation between the maternal and offspring genomes. In contrast, the association between prenatal smoking and offspring antisocial behavior depended on inherited factors because association was only present in related mothers and offspring. The results demonstrate that this unusual prenatal cross-fostering design is feasible and informative for disentangling inherited and prenatal effects on human health and behavior. Disentangling these different effects is invaluable for pinpointing markers of prenatal adversity that have a causal effect on offspring outcomes. The origins of behavior and many common complex disorders may begin in early life, therefore this experimental design could pave the way for identifying prenatal factors that affect behavior in future generations. PMID- 19188592 TI - Risk assessment in man and mouse. AB - Human and mouse subjects tried to anticipate at which of 2 locations a reward would appear. On a randomly scheduled fraction of the trials, it appeared with a short latency at one location; on the complementary fraction, it appeared after a longer latency at the other location. Subjects of both species accurately assessed the exogenous uncertainty (the probability of a short versus a long trial) and the endogenous uncertainty (from the scalar variability in their estimates of an elapsed duration) to compute the optimal target latency for a switch from the short- to the long-latency location. The optimal latency was arrived at so rapidly that there was no reliably discernible improvement over trials. Under these nonverbal conditions, humans and mice accurately assess risks and behave nearly optimally. That this capacity is well-developed in the mouse opens up the possibility of a genetic approach to the neurobiological mechanisms underlying risk assessment. PMID- 19188593 TI - Scoring diverse cellular morphologies in image-based screens with iterative feedback and machine learning. AB - Many biological pathways were first uncovered by identifying mutants with visible phenotypes and by scoring every sample in a screen via tedious and subjective visual inspection. Now, automated image analysis can effectively score many phenotypes. In practical application, customizing an image-analysis algorithm or finding a sufficient number of example cells to train a machine learning algorithm can be infeasible, particularly when positive control samples are not available and the phenotype of interest is rare. Here we present a supervised machine learning approach that uses iterative feedback to readily score multiple subtle and complex morphological phenotypes in high-throughput, image-based screens. First, automated cytological profiling extracts hundreds of numerical descriptors for every cell in every image. Next, the researcher generates a rule (i.e., classifier) to recognize cells with a phenotype of interest during a short, interactive training session using iterative feedback. Finally, all of the cells in the experiment are automatically classified and each sample is scored based on the presence of cells displaying the phenotype. By using this approach, we successfully scored images in RNA interference screens in 2 organisms for the prevalence of 15 diverse cellular morphologies, some of which were previously intractable. PMID- 19188594 TI - Conditional deletion of the MHC class I-related receptor FcRn reveals the sites of IgG homeostasis in mice. AB - The MHC class I-related receptor FcRn regulates the levels and persistence of IgG in vivo. This receptor salvages IgG from lysosomal degradation within cells, and the binding properties of an IgG for FcRn correlate with in vivo half-life. FcRn is expressed at multiple different sites throughout adult life. However, the cell types and sites at which FcRn maintains IgG homeostasis are not well defined. Toward understanding the sites of FcRn function, we have generated a mouse strain in which this Fc receptor can be conditionally deleted. In combination with mice that express Cre recombinase under the control of the Tie2 promoter (Tie2-Cre), the effect of site-specific deletion of floxed FcRn in endothelial and hematopoietic cells on IgG persistence was analyzed. The pharmacokinetics and steady-state levels of IgG in Tie2-Cre mice that are homozygous for the floxed FcRn allele reveal a complete loss of FcRn function in regulating the half-lives of wild-type IgG. The primary sites for the maintenance of endogenous IgGs in mice are therefore endothelial and hematopoietic cells. PMID- 19188595 TI - Genome-wide association analysis reveals a SOD1 mutation in canine degenerative myelopathy that resembles amyotrophic lateral sclerosis. AB - Canine degenerative myelopathy (DM) is a fatal neurodegenerative disease prevalent in several dog breeds. Typically, the initial progressive upper motor neuron spastic and general proprioceptive ataxia in the pelvic limbs occurs at 8 years of age or older. If euthanasia is delayed, the clinical signs will ascend, causing flaccid tetraparesis and other lower motor neuron signs. DNA samples from 38 DM-affected Pembroke Welsh corgi cases and 17 related clinically normal controls were used for genome-wide association mapping, which produced the strongest associations with markers on CFA31 in a region containing the canine SOD1 gene. SOD1 was considered a regional candidate gene because mutations in human SOD1 can cause amyotrophic lateral sclerosis (ALS), an adult-onset fatal paralytic neurodegenerative disease with both upper and lower motor neuron involvement. The resequencing of SOD1 in normal and affected dogs revealed a G to A transition, resulting in an E40K missense mutation. Homozygosity for the A allele was associated with DM in 5 dog breeds: Pembroke Welsh corgi, Boxer, Rhodesian ridgeback, German Shepherd dog, and Chesapeake Bay retriever. Microscopic examination of spinal cords from affected dogs revealed myelin and axon loss affecting the lateral white matter and neuronal cytoplasmic inclusions that bind anti-superoxide dismutase 1 antibodies. These inclusions are similar to those seen in spinal cord sections from ALS patients with SOD1 mutations. Our findings identify canine DM to be the first recognized spontaneously occurring animal model for ALS. PMID- 19188596 TI - Ocean acidification impairs olfactory discrimination and homing ability of a marine fish. AB - The persistence of most coastal marine species depends on larvae finding suitable adult habitat at the end of an offshore dispersive stage that can last weeks or months. We tested the effects that ocean acidification from elevated levels of atmospheric carbon dioxide (CO(2)) could have on the ability of larvae to detect olfactory cues from adult habitats. Larval clownfish reared in control seawater (pH 8.15) discriminated between a range of cues that could help them locate reef habitat and suitable settlement sites. This discriminatory ability was disrupted when larvae were reared in conditions simulating CO(2)-induced ocean acidification. Larvae became strongly attracted to olfactory stimuli they normally avoided when reared at levels of ocean pH that could occur ca. 2100 (pH 7.8) and they no longer responded to any olfactory cues when reared at pH levels (pH 7.6) that might be attained later next century on a business-as-usual carbon dioxide emissions trajectory. If acidification continues unabated, the impairment of sensory ability will reduce population sustainability of many marine species, with potentially profound consequences for marine diversity. PMID- 19188597 TI - Prevention of autoimmune disease by induction of tolerance to Toll-like receptor 7. AB - Activation of Toll-like receptors (TLR) contributes to the initiation and maintenance of chronic inflammation in autoimmune diseases, yet repeated exposure to a TLR agonist can induce hyporesponsiveness to subsequent TLR stimulation. Here, we used a synthetic TLR7 agonist, 9-benzyl-8-hydroxy-2-(2-methoxyethoxy) adenine (SM360320, 1V136) to study TLR7 induced attenuation of inflammatory responses and its application to autoimmune diseases. Repeated low dose administration of this TLR7 agonist induced hyporesponsiveness or tolerance to TLR2, -7, and -9 activators and limited the course of neural inflammation in an experimental allergic encephalomyelitis model. The hyporesponsiveness did not depend on T or B lymphocytes, but did require bone marrow derived cells. In addition, TLR7 tolerance reduced inflammation in a passive antibody mediated arthritis model. TLR7 tolerance did not cause global immunosuppression, because susceptibility to Listeria monocytogenes infection was not altered. The mechanism of TLR7 tolerance involved the up-regulation of 2 inhibitors of TLR signaling: Interleukin 1 Receptor Associated Kinase (IRAK) M, and Src homology 2 domain containing inositol polyphosphate phosphatase (SHIP)-1. These findings suggest that induction of TLR7 tolerance might be a new therapeutic approach to subdue inflammation in autoimmune diseases. PMID- 19188599 TI - Defined tuberculosis vaccine, Mtb72F/AS02A, evidence of protection in cynomolgus monkeys. AB - The development of a vaccine for tuberculosis requires a combination of antigens and adjuvants capable of inducing appropriate and long-lasting T cell immunity. We evaluated Mtb72F formulated in AS02A in the cynomolgus monkey model. The vaccine was immunogenic and caused no adverse reactions. When monkeys were immunized with bacillus Calmette-Guerin (BCG) and then boosted with Mtb72F in AS02A, protection superior to that afforded by using BCG alone was achieved, as measured by clinical parameters, pathology, and survival. We observed long-term survival and evidence of reversal of disease progression in monkeys immunized with the prime-boost regimen. Antigen-specific responses from protected monkeys receiving BCG and Mtb72F/AS02A had a distinctive cytokine profile characterized by an increased ratio between 3 Th1 cytokines, IFN-gamma, TNF, and IL-2 and an innate cytokine, IL-6. To our knowledge, this is an initial report of a vaccine capable of inducing long-term protection against tuberculosis in a nonhuman primate model, as determined by protection against severe disease and death, and by other clinical and histopathological parameters. PMID- 19188598 TI - Female rats learn trace memories better than male rats and consequently retain a greater proportion of new neurons in their hippocampi. AB - Learning increases the survival of new cells that are generated in the hippocampal formation before the training experience, especially if the animal learns to associate stimuli across time [Gould E, Beylin A, Tanapat P, Reeves A, Shors TJ (1999) Nat Neurosci 2:260-265]. All relevant studies have been conducted on male rats, despite evidence for sex differences in this type of learning. In the present study, we asked whether sex differences in learning influence the survival of neurons generated in the adult hippocampus. Male and female adult rats were injected with one dose of bromodeoxyuridine (BrdU; 200 mg/kg), to label one population of dividing cells. One week later, half of the animals were trained with a temporal learning task of trace eyeblink conditioning, while the other half were not trained. Animals were killed 1 day after training (12 days after the BrdU injection). Hippocampal tissue was stained for BrdU and a marker of immature neurons, doublecortin. Both sexes learned to emit the conditioned eyeblink response during the trace interval. As a consequence, more new neurons remained in their hippocampi than in sex-matched controls. In individual animals, the number of surviving cells correlated positively with asymptotic performance; those that expressed more learned responses retained more new neurons. However, animals that learned very well retained even more new cells if they required many trials to do so. Because females emitted more learned responses than males did, they retained nearly twice as many new cells per unit volume of tissue. This effect was most evident in the ventral region of the hippocampal formation. Thus, sex differences in learning alter the anatomical structure of the hippocampus. As a result, male and female brains continue to differentiate in adulthood. PMID- 19188600 TI - The transactivating function 1 of estrogen receptor alpha is dispensable for the vasculoprotective actions of 17beta-estradiol. AB - Full-length 66-kDa estrogen receptor alpha (ERalpha) stimulates target gene transcription through two activation functions (AFs), AF-1 in the N-terminal domain and AF-2 in the ligand binding domain. Another physiologically expressed 46-kDa ERalpha isoform lacks the N-terminal A/B domains and is consequently devoid of AF-1. Previous studies in cultured endothelial cells showed that the N terminal A/B domain might not be required for estradiol (E2)-elicited NO production. To evaluate the involvement of ERalpha AF-1 in the vasculoprotective actions of E2, we generated a targeted deletion of the ERalpha A/B domain in the mouse. In these ERalphaAF-1(0) mice, both basal endothelial NO production and reendothelialization process were increased by E2 administration to a similar extent than in control mice. Furthermore, exogenous E2 similarly decreased fatty streak deposits at the aortic root from both ovariectomized 18-week-old ERalphaAF 1(+/+) LDLr(-/-) (low-density lipoprotein receptor) and ERalphaAF-1(0) LDLr (-/-) mice fed with a hypercholesterolemic diet. In addition, quantification of lesion size on en face preparations of the aortic tree of 8-month-old ovariectomized or intact female mice revealed that ERalpha AF-1 is dispensable for the atheroprotective action of endogenous estrogens. We conclude that ERalpha AF-1 is not required for three major vasculoprotective actions of E2, whereas it is necessary for the effects of E2 on its reproductive targets. Thus, selective ER modulators stimulating ERalpha with minimal activation of ERalpha AF-1 could retain beneficial vascular actions, while minimizing the sexual effects. PMID- 19188601 TI - Predicting human resting-state functional connectivity from structural connectivity. AB - In the cerebral cortex, the activity levels of neuronal populations are continuously fluctuating. When neuronal activity, as measured using functional MRI (fMRI), is temporally coherent across 2 populations, those populations are said to be functionally connected. Functional connectivity has previously been shown to correlate with structural (anatomical) connectivity patterns at an aggregate level. In the present study we investigate, with the aid of computational modeling, whether systems-level properties of functional networks- including their spatial statistics and their persistence across time--can be accounted for by properties of the underlying anatomical network. We measured resting state functional connectivity (using fMRI) and structural connectivity (using diffusion spectrum imaging tractography) in the same individuals at high resolution. Structural connectivity then provided the couplings for a model of macroscopic cortical dynamics. In both model and data, we observed (i) that strong functional connections commonly exist between regions with no direct structural connection, rendering the inference of structural connectivity from functional connectivity impractical; (ii) that indirect connections and interregional distance accounted for some of the variance in functional connectivity that was unexplained by direct structural connectivity; and (iii) that resting-state functional connectivity exhibits variability within and across both scanning sessions and model runs. These empirical and modeling results demonstrate that although resting state functional connectivity is variable and is frequently present between regions without direct structural linkage, its strength, persistence, and spatial statistics are nevertheless constrained by the large-scale anatomical structure of the human cerebral cortex. PMID- 19188602 TI - MENepsilon/beta noncoding RNAs are essential for structural integrity of nuclear paraspeckles. AB - Recent transcriptome analyses have shown that thousands of noncoding RNAs (ncRNAs) are transcribed from mammalian genomes. Although the number of functionally annotated ncRNAs is still limited, they are known to be frequently retained in the nucleus, where they coordinate regulatory networks of gene expression. Some subnuclear organelles or nuclear bodies include RNA species whose identity and structural roles are largely unknown. We identified 2 abundant overlapping ncRNAs, MENepsilon and MENbeta (MENepsilon/beta), which are transcribed from the corresponding site in the multiple endocrine neoplasia (MEN) I locus and which localize to nuclear paraspeckles. This finding raises the intriguing possibility that MENepsilon/beta are involved in paraspeckle organization, because paraspeckles are, reportedly, RNase-sensitive structures. Successful removal of MENepsilon/beta by a refined knockdown method resulted in paraspeckle disintegration. Furthermore, the reassembly of paraspeckles disassembled by transcriptional arrest appeared to be unsuccessful in the absence of MENepsilon/beta. RNA interference and immunoprecipitation further revealed that the paraspeckle proteins p54/nrb and PSF selectively associate with and stabilize the longer MENbeta, thereby contributing to the organization of the paraspeckle structure. The paraspeckle protein PSP1 is not directly involved in either MENepsilon/beta stabilization or paraspeckle organization. We postulate a model for nuclear paraspeckle body organization where specific ncRNAs and RNA binding proteins cooperate to maintain and, presumably, establish the structure. PMID- 19188603 TI - Gastrointestinal differentiation marker Cytokeratin 20 is regulated by homeobox gene CDX1. AB - CDX1 is a transcription factor that plays a key role in intestinal development and differentiation. However, the downstream targets of CDX1 are less well defined than those of its close homologue, CDX2. We report here the identification of downstream targets of CDX1 using microarray gene-expression analysis and other approaches. Keratin 20 (KRT20), a member of the intermediate filament and a well-known marker of intestinal differentiation, was initially identified as one of the genes likely to be directly regulated by CDX1. CDX1 and KRT20 mRNA expression were significantly correlated in a panel of 38 colorectal cancer cell lines. Deletion and mutation analysis of the KRT20 promoter showed that the minimum regulatory region for the control of KRT20 expression by CDX1 is within 246 bp upstream of the KRT20 transcription start site. ChIP analysis confirmed that CDX1 binds to the predicted CDX elements in this region of the KRT20 promoter in vivo. In addition, immunohistochemistry showed expression of CDX1 parallels that of KRT20 in the normal crypt, which further supports their close relationship. In summary, our observations strongly imply that KRT20 is directly regulated by CDX1, and therefore suggest a role for CDX1 in maintaining differentiation in intestinal epithelial cells. Because a key feature of the development of a cancer is an unbalanced program of proliferation and differentiation, dysregulation of CDX1 may be an advantage for the development of a colorectal carcinoma. This could, therefore, explain the relatively frequent down regulation of CDX1 in colorectal carcinomas by hypermethylation. PMID- 19188604 TI - Early childhood stress is associated with elevated antibody levels to herpes simplex virus type 1. AB - It is well known that children need solicitous parenting and a nurturing rearing environment to ensure their normal behavioral development. Early adversity often negatively impacts emotional and mental well-being, but it is less clearly established how much the maturation and regulation of physiological systems is also compromised. The following research investigated the effect of 2 different types of adverse childhood experiences, early deprivation through institutionalization and physical abuse, on a previously unexplored outcome: the containment of herpes simplex virus (HSV). The presence of HSV-specific antibody in salivary specimens was determined in 155 adolescents, including 41 postinstitutionalized, 34 physically-abused, and 80 demographically-similar control youth. Across 4 school and home days, HSV antibody was higher in both postinstitutionalized and physically-abused adolescents when compared with control participants. Because the prevalence of HSV infection was similar across the groups, the elevated antibody was likely indicative of viral recrudescence from latency. Total secretory Ig-A secretion was associated with HSV, but did not account for the group differences in HSV-specific antibody. These findings are likely caused by a failure of cellular immune processes to limit viral reactivation, indicating a persistent effect of early rearing on immune functioning. The fact that antibody profiles were still altered years after adoption into a more benevolent setting with supportive families suggests these results were not caused by contemporaneous factors, but rather reflect a lingering influence of earlier life experiences. PMID- 19188605 TI - Evidence of cacao use in the Prehispanic American Southwest. AB - Chemical analyses of organic residues in fragments of ceramic vessels from Pueblo Bonito in Chaco Canyon, New Mexico, reveal theobromine, a biomarker for cacao. With an estimated 800 rooms, Pueblo Bonito is the largest archaeological site in Chaco Canyon and was the center of a large number of interconnected towns and villages spread over northwestern New Mexico. The cacao residues come from pieces of vessels that are likely cylinder jars, special containers occurring almost solely at Pueblo Bonito and deposited in caches at the site. This first known use of cacao drinks north of the Mexican border indicates exchange with cacao cultivators in Mesoamerica in a time frame of about A.D. 1000-1125. The association of cylinder jars and cacao beverages suggests that the Chacoan ritual involving the drinking of cacao was tied to Mesoamerican rituals incorporating cylindrical vases and cacao. The importance of Pueblo Bonito within the Chacoan world likely lies in part with the integration of Mesoamerican ritual, including critical culinary ingredients. PMID- 19188606 TI - Alignment-free genome comparison with feature frequency profiles (FFP) and optimal resolutions. AB - For comparison of whole-genome (genic + nongenic) sequences, multiple sequence alignment of a few selected genes is not appropriate. One approach is to use an alignment-free method in which feature (or l-mer) frequency profiles (FFP) of whole genomes are used for comparison-a variation of a text or book comparison method, using word frequency profiles. In this approach it is critical to identify the optimal resolution range of l-mers for the given set of genomes compared. The optimum FFP method is applicable for comparing whole genomes or large genomic regions even when there are no common genes with high homology. We outline the method in 3 stages: (i) We first show how the optimal resolution range can be determined with English books which have been transformed into long character strings by removing all punctuation and spaces. (ii) Next, we test the robustness of the optimized FFP method at the nucleotide level, using a mutation model with a wide range of base substitutions and rearrangements. (iii) Finally, to illustrate the utility of the method, phylogenies are reconstructed from concatenated mammalian intronic genomes; the FFP derived intronic genome topologies for each l within the optimal range are all very similar. The topology agrees with the established mammalian phylogeny revealing that intron regions contain a similar level of phylogenic signal as do coding regions. PMID- 19188607 TI - The feeding biomechanics and dietary ecology of Australopithecus africanus. AB - The African Plio-Pleistocene hominins known as australopiths evolved a distinctive craniofacial morphology that traditionally has been viewed as a dietary adaptation for feeding on either small, hard objects or on large volumes of food. A historically influential interpretation of this morphology hypothesizes that loads applied to the premolars during feeding had a profound influence on the evolution of australopith craniofacial form. Here, we test this hypothesis using finite element analysis in conjunction with comparative, imaging, and experimental methods. We find that the facial skeleton of the Australopithecus type species, A. africanus, is well suited to withstand premolar loads. However, we suggest that the mastication of either small objects or large volumes of food is unlikely to fully explain the evolution of facial form in this species. Rather, key aspects of australopith craniofacial morphology are more likely to be related to the ingestion and initial preparation of large, mechanically protected food objects like large nuts and seeds. These foods may have broadened the diet of these hominins, possibly by being critical resources that australopiths relied on during periods when their preferred dietary items were in short supply. Our analysis reconciles apparent discrepancies between dietary reconstructions based on biomechanics, tooth morphology, and dental microwear. PMID- 19188608 TI - Substitutional alloy of Ce and Al. AB - The formation of substitutional alloys has been restricted to elements with similar atomic radii and electronegativity. Using high-pressure at 298 K, we synthesized a face-centered cubic disordered alloy of highly dissimilar elements (large Ce and small Al atoms) by compressing the Ce(3)Al intermetallic compound >15 GPa or the Ce(3)Al metallic glass >25 GPa. Synchrotron X-ray diffraction, Ce L(3)-edge absorption spectroscopy, and ab initio calculations revealed that the pressure-induced Kondo volume collapse and 4f electron delocalization of Ce reduced the differences between Ce and Al and brought them within the Hume Rothery (HR) limit for substitutional alloying. The alloy remained after complete release of pressure, which was also accompanied by the transformation of Ce back to its ambient 4f electron localized state and reversal of the Kondo volume collapse, resulting in a non-HR alloy at ambient conditions. PMID- 19188610 TI - From the Cover: Strong profiling is not mathematically optimal for discovering rare malfeasors. AB - The use of profiling by ethnicity or nationality to trigger secondary security screening is a controversial social and political issue. Overlooked is the question of whether such actuarial methods are in fact mathematically justified, even under the most idealized assumptions of completely accurate prior probabilities, and secondary screenings concentrated on the highest-probability individuals. We show here that strong profiling (defined as screening at least in proportion to prior probability) is no more efficient than uniform random sampling of the entire population, because resources are wasted on the repeated screening of higher probability, but innocent, individuals. A mathematically optimal strategy would be "square-root biased sampling," the geometric mean between strong profiling and uniform sampling, with secondary screenings distributed broadly, although not uniformly, over the population. Square-root biased sampling is a general idea that can be applied whenever a "bell-ringer" event must be found by sampling with replacement, but can be recognized (either with certainty, or with some probability) when seen. PMID- 19188609 TI - Protection of synapses against Alzheimer's-linked toxins: insulin signaling prevents the pathogenic binding of Abeta oligomers. AB - Synapse deterioration underlying severe memory loss in early Alzheimer's disease (AD) is thought to be caused by soluble amyloid beta (Abeta) oligomers. Mechanistically, soluble Abeta oligomers, also referred to as Abeta-derived diffusible ligands (ADDLs), act as highly specific pathogenic ligands, binding to sites localized at particular synapses. This binding triggers oxidative stress, loss of synaptic spines, and ectopic redistribution of receptors critical to plasticity and memory. We report here the existence of a protective mechanism that naturally shields synapses against ADDL-induced deterioration. Synapse pathology was investigated in mature cultures of hippocampal neurons. Before spine loss, ADDLs caused major downregulation of plasma membrane insulin receptors (IRs), via a mechanism sensitive to calcium calmodulin-dependent kinase II (CaMKII) and casein kinase II (CK2) inhibition. Most significantly, this loss of surface IRs, and ADDL-induced oxidative stress and synaptic spine deterioration, could be completely prevented by insulin. At submaximal insulin doses, protection was potentiated by rosiglitazone, an insulin-sensitizing drug used to treat type 2 diabetes. The mechanism of insulin protection entailed a marked reduction in pathogenic ADDL binding. Surprisingly, insulin failed to block ADDL binding when IR tyrosine kinase activity was inhibited; in fact, a significant increase in binding was caused by IR inhibition. The protective role of insulin thus derives from IR signaling-dependent downregulation of ADDL binding sites rather than ligand competition. The finding that synapse vulnerability to ADDLs can be mitigated by insulin suggests that bolstering brain insulin signaling, which can decline with aging and diabetes, could have significant potential to slow or deter AD pathogenesis. PMID- 19188612 TI - Spatiotemporal structures in aging and rejuvenating glasses. AB - Complex spatiotemporal structures develop during the process of aging glasses after cooling and of rejuvenating glasses on heating. The key to understanding these structures is the interplay between the activated reconfiguration events that generate mobility and the transport of mobility. These effects are both accounted for by combining the random first-order transition theory of activated events with mode coupling theory in an inhomogeneous setting. The predicted modifications by mobility transport of the time course of the aging regime are modest. In contrast, the rejuvenation process is strongly affected through the propagation of fronts of enhanced mobility originating from the initial reconfiguration events. The structures in a rejuvenating glass resemble flames. An analysis along the lines of combustion theory provides an estimate of the front propagation speed. Heterogeneous rejuvenation naturally should occur for glasses with free surfaces. The analogy with combustion also provides a way of looking at the uptake of diluents by glasses described by case II and super case II diffusion. PMID- 19188613 TI - Proxy inconsistency and other problems in millennial paleoclimate reconstructions. PMID- 19188611 TI - Melanin-concentrating hormone neurons discharge in a reciprocal manner to orexin neurons across the sleep-wake cycle. AB - Neurons containing melanin-concentrating hormone (MCH) are codistributed with neurons containing orexin (Orx or hypocretin) in the lateral hypothalamus, a peptide and region known to be critical for maintaining wakefulness. Evidence from knockout and c-Fos studies suggests, however, that the MCH neurons might play a different role than Orx neurons in regulating activity and sleep-wake states. To examine this possibility, neurons were recorded across natural sleep wake states in head-fixed rats and labeled by using the juxtacellular technique for subsequent immunohistochemical identification. Neurons identified as MCH+ did not fire during wake (W); they fired selectively during sleep, occasionally during slow wave sleep (SWS) and maximally during paradoxical sleep (PS). As W Off/Sleep-On, the MCH neurons discharged in a reciprocal manner to the W-On/Sleep Off Orx neurons and could accordingly play a complementary role to Orx neurons in sleep-wake state regulation and contribute to the pathophysiology of certain sleep disorders, such as narcolepsy with cataplexy. PMID- 19188615 TI - Get excess salt out of our diet. PMID- 19188616 TI - Multidrug-resistant Neisseria gonorrhoeae. PMID- 19188617 TI - Proper studies of selective serotonin reuptake inhibitors are needed for youth with depression. PMID- 19188619 TI - Rapidly rising clinical trial costs worry researchers. PMID- 19188618 TI - Transparency in the delivery of lawful abortion services. PMID- 19188620 TI - Drug development cost estimates hard to swallow. PMID- 19188621 TI - Public health insurance in Uganda still only a dream. PMID- 19188622 TI - Legal consensus eludes Europe. PMID- 19188624 TI - Trends in the profile of the Canadian physician pool. PMID- 19188626 TI - Prevalence of and risk factors for quinolone-resistant Neisseria gonorrhoeae infection in Ontario. AB - BACKGROUND: Quinolone-resistant Neisseria gonorrhoeae has swiftly emerged in Canada. We sought to determine its prevalence in the province of Ontario and to investigate risk factors for quinolone-resistant N. gonorrhoeae infection in a Canadian setting. METHODS: We used records from the Public Health Laboratory of the Ontario Agency for Health Protection and Promotion in Toronto, Ontario, and the National Microbiology Laboratory in Winnipeg, Manitoba, to generate epidemic curves for N. gonorrhoeae infection. We extracted limited demographic data from 2006 quinolone-resistant N. gonorrhoeae isolates and from a random sample of quinolone-susceptible isolates. We also extracted minimum inhibitory concentrations for commonly tested antibiotics. RESULTS: Between 2002 and 2006, the number of N. gonorrhoeae infections detected by culture decreased by 26% and the number of cases detected by nucleic acid amplification testing increased 6 fold. The proportion of N. gonorrhoeae isolates with resistance to quinolones increased from 4% to 28% over the same period. Analysis of 695 quinolone resistant N. gonorrhoeae isolates and 688 quinolone-susceptible control isolates from 2006 showed a higher proportion of men (odds ratio [OR] 3.1, 95% confidence interval [CI] 2.3-4.1) and patients over 30 years of age (OR 3.1, 95% CI 2.4-3.8) in the quinolone-resistant group. The proportion of men who have sex with men appeared to be relatively similar in both groups (OR 1.4, 95% CI 1.1-1.8). Quinolone-resistant strains were more resistant to penicillin (p < 0.001), tetracycline (p < 0.001) and erythromycin (p < 0.001). All isolates were susceptible to cefixime, ceftriaxone, azithromycin and spectinomycin. INTERPRETATION: During 2006 in Ontario, 28% of N. gonorrhoeae isolates were resistant to quinolones. Infections in heterosexual men appear to have contributed significantly to the quinolone resistance rate. Medical practitioners should be aware of the widespread prevalence of quinolone-resistant N. gonorrhoeae and avoid quinolone use for empiric therapy. PMID- 19188627 TI - Selective serotonin reuptake inhibitors and risk of suicide: a systematic review of observational studies. AB - BACKGROUND: It is unclear whether the use of selective serotonin reuptake inhibitors (SSRIs) and other antidepressant drugs reduce the risk of suicide in people with depression. We explored the association between exposure to SSRIs and risk of suicide completion or attempt. METHODS: We conducted a systematic review of observational studies that reported completed or attempted suicide in depressed individuals who were exposed to SSRIs compared with those who were not exposed to antidepressants. We assessed the overall risk of completed or attempted suicide. RESULTS: Eight studies involving more than 200 000 patients with moderate or severe depression were included in the meta-analysis. Although exposure to SSRIs increased the risk of completed or attempted suicide among adolescents (odds ratio [OR] 1.92, 95% confidence interval [CI] 1.51-2.44), the risk was decreased among adults (OR 0.57, 95% CI 0.47-0.70). Among people aged 65 or more years, exposure to SSRIs had a protective effect (OR 0.46, 95% CI 0.27 0.79). Sensitivity analyses did not change these findings. In particular, for studies that used completed suicide as an outcome, exposure to SSRIs was associated with increased risk among adolescents (OR 5.81, 95% CI 1.57-21.51) and decreased risk among adults (OR 0.66, 95% CI 0.52-0.83) and older people (OR 0.53, 95% CI 0.26-1.06). INTERPRETATION: Based on data from observational studies, use of SSRIs may be associated with a reduced risk of suicide in adults with depression. Among adolescents, use of SSRIs may increase suicidality. PMID- 19188628 TI - Clandestine induced abortion: prevalence, incidence and risk factors among women in a Latin American country. AB - BACKGROUND: Clandestine induced abortions are a public health problem in many developing countries where access to abortion services is legally restricted. We estimated the prevalence and incidence of, and risk factors for, clandestine induced abortions in a Latin American country. METHODS: We conducted a large population-based survey of women aged 18-29 years in 20 cities in Peru. We asked questions about their history of spontaneous and induced abortions, using techniques to encourage disclosure. RESULTS: Of 8242 eligible women, 7992 (97.0%) agreed to participate. The prevalence of reported induced abortions was 11.6% (95% confidence interval [CI] 10.9%-12.4%) among the 7962 women who participated in the survey. It was 13.6% (95% CI 12.8%-14.5%) among the 6559 women who reported having been sexually active. The annual incidence of induced abortion was 3.1% (95% CI 2.9%-3.3%) among the women who had ever been sexually active. In the multivariable analysis, risk factors for induced abortion were higher age at the time of the survey (odds ratio [OR] 1.11, 95% CI 1.07-1.15), lower age at first sexual intercourse (OR 0.87, 95% CI 0.84-0.91), geographic region (highlands: OR 1.56, 95% CI 1.23-1.97; jungle: OR 1.81, 95% CI 1.41-2.31 [v. coastal region]), having children (OR 0.82, 95% CI 0.68-0.98), having more than 1 sexual partner in lifetime (2 partners: OR 1.61, 95% CI 1.23-2.09; > or = 3 partners: OR 2.79, 95% CI 2.12-3.67), and having 1 or more sexual partners in the year before the survey (1 partner: OR 1.36, 95% CI 1.01-1.72; > or = 2 partners: OR 1.54, 95% CI 1.14-2.02). Overall, 49.0% (95% CI 47.6%-50.3%) of the women who reported being currently sexually active were not using contraception. INTERPRETATION: The incidence of clandestine, potentially unsafe induced abortion in Peru is as high as or higher than the rates in many countries where induced abortion is legal and safe. The provision of contraception and safer-sex education to those who require it needs to be greatly improved and could potentially reduce the rate of induced abortion. PMID- 19188630 TI - Cuba's system of maternal health and early childhood development: lessons for Canada. PMID- 19188629 TI - Neurobiological mechanisms in major depressive disorder. PMID- 19188631 TI - Abdominal aortic pseudocoarctation. PMID- 19188632 TI - Pneumococcal vaccination and myocardial infarction. PMID- 19188633 TI - Pneumococcal vaccination and myocardial infarction. PMID- 19188638 TI - Superior vena cava syndrome. PMID- 19188639 TI - Chief thieves. PMID- 19188640 TI - Update on the State Children's Health Insurance Program. PMID- 19188642 TI - Minimal risk in pediatric research as a function of age. AB - The minimal risk standard allows institutional review boards (IRBs) to approve pediatric research when the risks do not exceed the risks children face in daily life or during routine examinations. The fact that the risks children face in daily life vary with age raises the question of whether IRBs should implement this standard by adopting 1, 2, 3, or even more risk standards. The level of research risks to which it is appropriate to expose children depends on their maturity level. While younger children should be exposed to only very low research risks, it can be appropriate to expose older children to somewhat higher risks. This analysis supports 2 thresholds for minimal risk in pediatric research: one for research with younger children and a distinct standard for older children who can understand and who agree to participate. PMID- 19188641 TI - Racial differences in parents' distrust of medicine and research. AB - OBJECTIVE: To assess and compare the attitudes and trust that African American and white parents have toward their children participating in research. DESIGN: Self-administered, cross-sectional survey of a convenience sample of parents. SETTING: Primary Care Center at Children's Hospital of Pittsburgh from August 2004 through April 2005. PARTICIPANTS: One hundred ninety parents (140 African American and 50 white parents). OUTCOME MEASURE: Parental distrust of medical research as measured by a summative score of distrusting responses to 8 questions assessing trust in research. RESULTS: African American parents had significantly greater distrust than white parents (67% vs 50%, P = .04). Education was also associated with having significantly greater distrust (74% of those with or = 3 pulmonary hemorrhage rates were < or = 1.5% for all arms despite 9% of patients receiving therapeutic anticoagulation. CONCLUSION: Combining bevacizumab (7.5 or 15 mg/kg) with CG significantly improved PFS and objective response rate. Bevacizumab plus platinum-based chemotherapy offers clinical benefit for bevacizumab-eligible patients with advanced NSCLC. PMID- 19188681 TI - Highly purified Th17 cells from BDC2.5NOD mice convert into Th1-like cells in NOD/SCID recipient mice. AB - Th17 cells are involved in the pathogenesis of many autoimmune diseases, but it is not clear whether they play a pathogenic role in type 1 diabetes. Here we investigated whether mouse Th17 cells with specificity for an islet antigen can induce diabetes upon transfer into NOD/SCID recipient mice. Induction of diabetes in NOD/SCID mice via adoptive transfer of Th1 cells from BDC2.5 transgenic mice was prevented by treatment of the recipient mice with a neutralizing IFN-gamma specific antibody. This result suggested a major role of Th1 cells in the induction of disease in this model of type 1 diabetes. Nevertheless, transfer of highly purified Th17 cells from BDC2.5 transgenic mice caused diabetes in NOD/SCID recipients with similar rates of onset as in transfer of Th1 cells. However, treatment with neutralizing IL-17-specific antibodies did not prevent disease. Instead, the transferred Th17 cells, completely devoid of IFN-gamma at the time of transfer, rapidly converted to secrete IFN-gamma in the NOD/SCID recipients. Purified Th17 cells also upregulated Tbet and secreted IFN-gamma upon exposure to IL-12 in vitro and in vivo in NOD/SCID recipients. These results indicate substantial plasticity of Th17 commitment toward a Th1-like profile. PMID- 19188682 TI - Macrophage-derived human resistin exacerbates adipose tissue inflammation and insulin resistance in mice. AB - Resistin is an adipokine that contributes to insulin resistance in mice. In humans, however, studies investigating the link between resistin and metabolic disease are conflicting. Further complicating the matter, human resistin is produced mainly by macrophages rather than adipocytes. To address this important issue, we generated mice that lack adipocyte-derived mouse resistin but produce human resistin in a pattern similar to that found in humans, i.e., in macrophages (humanized resistin mice). When placed on a high-fat diet, the humanized resistin mice rapidly developed accelerated white adipose tissue (WAT) inflammation, leading to increased lipolysis and increased serum free fatty acids. Over time, these mice accumulated lipids, including diacylglycerols, in muscle. We found that this resulted in increased Pkcq pathway activity, leading to increased serine phosphorylation of Irs-1 and insulin resistance. Thus, although the site of resistin production differs between species, human resistin exacerbates WAT inflammation and contributes to insulin resistance. PMID- 19188683 TI - Mitochondrial H2O2 emission and cellular redox state link excess fat intake to insulin resistance in both rodents and humans. AB - High dietary fat intake leads to insulin resistance in skeletal muscle, and this represents a major risk factor for type 2 diabetes and cardiovascular disease. Mitochondrial dysfunction and oxidative stress have been implicated in the disease process, but the underlying mechanisms are still unknown. Here we show that in skeletal muscle of both rodents and humans, a diet high in fat increases the H(2)O(2)-emitting potential of mitochondria, shifts the cellular redox environment to a more oxidized state, and decreases the redox-buffering capacity in the absence of any change in mitochondrial respiratory function. Furthermore, we show that attenuating mitochondrial H(2)O(2) emission, either by treating rats with a mitochondrial-targeted antioxidant or by genetically engineering the overexpression of catalase in mitochondria of muscle in mice, completely preserves insulin sensitivity despite a high-fat diet. These findings place the etiology of insulin resistance in the context of mitochondrial bioenergetics by demonstrating that mitochondrial H(2)O(2) emission serves as both a gauge of energy balance and a regulator of cellular redox environment, linking intracellular metabolic balance to the control of insulin sensitivity. PMID- 19188684 TI - Somatic mutation and functional polymorphism of a novel regulatory element in the HGF gene promoter causes its aberrant expression in human breast cancer. AB - The HGF gene is transcriptionally silenced in normal differentiated breast epithelial cells, but its repression fails to occur in mammary carcinoma tissues and cell lines. The molecular mechanisms underpinning aberrant HGF expression in breast cancer cells are unknown. Here we report the discovery of a DNA element located 750 bp upstream from the transcription start site in the human HGF promoter that acts as a transcriptional repressor and is a target of deletion mutagenesis in human breast cancer cells and tissues. This HGF promoter element consists of a mononucleotide repeat of 30 deoxyadenosines (30As), which we have termed "deoxyadenosine tract element" (DATE). Functional studies revealed that truncation mutations within DATE have profound local and global effects on the HGF promoter region by modulating chromatin structure and DNA-protein interactions, leading to constitutive activation of the HGF promoter in human breast carcinoma cell lines. We found that 51% of African Americans and 15% of individuals of mixed European descent with breast cancer harbor a truncated DATE variant (25As or fewer) in their breast tumors and that the truncated allele is associated with cancer incidence and aberrant HGF expression. Notably, breast cancer patients with the truncated DATE variant are substantially younger than those with a wild-type genotype. We also suggest that DATE may be used as a potential genetic marker to identify individuals with a higher risk of developing breast cancer. PMID- 19188686 TI - Telomeres and telomerase. Preface. PMID- 19188685 TI - Antioxidant or neurotrophic factor treatment preserves function in a mouse model of neovascularization-associated oxidative stress. AB - In several disease states, abnormal growth of blood vessels is associated with local neuronal degeneration. This is particularly true in ocular diseases such as retinal angiomatous proliferation (RAP) and macular telangiectasia (MacTel), in which, despite the absence of large-scale leakage or hemorrhage, abnormal neovascularization (NV) is associated with local neuronal dysfunction. We describe here a retinal phenotype in mice with dysfunctional receptors for VLDL (Vldlr-/- mice) that closely resembles human retinal diseases in which abnormal intra- and subretinal NV is associated with photoreceptor cell death. Such cell death was evidenced by decreased cone and, to a lesser extent, rod opsin expression and abnormal electroretinograms. Cell death in the region of intraretinal vascular abnormalities was associated with an increased presence of markers associated with oxidative stress. Oral antioxidant supplementation protected against photoreceptor degeneration and preserved retinal function, despite the continued presence of abnormal intra- and subretinal vessels. What we believe to be novel, Muller cell-based, virally mediated delivery of neurotrophic compounds specifically to sites of NV was also neuroprotective. These observations demonstrate that neuronal loss secondary to NV can be prevented by the use of simple antioxidant dietary measures or cell-based delivery of neurotrophic factors, even when the underlying vascular phenotype is not altered. PMID- 19188688 TI - Crosstalk between chromatin structure, nuclear compartmentalization, and telomere biology. AB - Telomeres are heterochromatic structures essential for the maintenance of genomic stability and the proliferative potential of human somatic cells. A minimal length of telomeric DNA repeats and proper binding of specialized proteins are required for the maintenance of telomere structure and function. Activation of telomere length maintenance mechanisms is considered a hallmark of cancer, while attrition of telomeres is a known contributor to the phenotypes associated with the aging process. However, the molecular mechanisms responsible for telomere homeostasis are not completely understood. Compelling data indicates that the epigenetic status of telomeric and subtelomeric chromatin also plays a role in the regulation of telomere biology. In addition, genomic regions are not randomly distributed within the nucleus, but rather compartmentalized into spatial and temporal domains whose functional implications for the cell are only beginning to be unraveled. Recent studies indicate that alterations in some of the mechanisms responsible for nuclear organization associate with defects in chromatin structure. These observations suggest a link between these two processes, raising the question of whether nuclear compartmentalization of telomeres might also impact on telomere biology. PMID- 19188687 TI - The telomeric transcriptome and SMG proteins at the crossroads. AB - Telomeres constitute the heterochromatic structures at the ends of linear eukaryotic chromosomes and are essential for the maintenance of genome stability. The longstanding belief that telomeres are transcriptionally silent has recently been overturned by the discovery of a telomeric transcriptome comprising telomeric repeat-containing RNA (TERRA) molecules. In mammalian cells, TERRA remains associated with telomeres after its transcription, and this association is regulated by SMG proteins, renowned for their involvement in several RNA metabolism pathways. The three SMG factors UPF1, hEST1A and SMG1, which have been associated with DNA stability networks before, also seem to be crucial to guarantee telomere integrity. Here, we review the current knowledge available of TERRA and on the role of SMG proteins in maintaining genome stability. We also propose an integrative model illustrating a putative link between telomere integrity, TERRA and SMG proteins. PMID- 19188689 TI - Cellular senescence, telomere recombination and maintenance. AB - Cellular senescence can be activated by various types of stressful stimuli, including telomere shortening, oncogenic or tumor suppressor signals, and DNA damage. Progressive telomere shortening in successive cell divisions induces senescence due to the loss of terminal sequences during DNA replication. Maintenance of the telomere sequences at human chromosome ends is essential for immortalized cells to escape from the normal limitations of the proliferation capacity. In this article, the molecular and functional details of telomere maintenance and cellular senescence are reviewed, including the signals that trigger senescence, telomere capping, and the telomere length maintenance mechanisms. PMID- 19188690 TI - Telomeric repeats far from the ends: mechanisms of origin and role in evolution. AB - In addition to their location at terminal positions, telomeric-like repeats are also present at internal sites of the chromosomes (intrachromosomal or interstitial telomeric sequences, ITSs). According to their sequence organization and genomic location, two different kinds of ITSs can be identified: (1) heterochromatic ITSs (het-ITSs), large (up to hundreds of kb) stretches of telomeric-like DNA localized mainly at centromeres, and (2) short ITSs (s-ITSs), short stretches of telomeric hexamers distributed at internal sites of the chromosomes. Het-ITSs have been only described in some vertebrate species and they probably represent the remnants of evolutionary chromosomal rearrangements. On the contrary, s-ITSs are probably present in all mammalian genomes although they have been described in detail only in some completely sequenced genomes. Sequence database analysis revealed the presence of 83, 79, 244 and 250 such s ITSs in the human, chimpanzee, mouse and rat genomes, respectively. Analysis of the flanking sequences suggested that s-ITSs were inserted during the repair of DNA double-strand breaks that occurred in the course of evolution. An extensive comparative analysis of the s-ITS loci and their orthologous 'empty' loci confirmed this hypothesis and suggested that the repair event involved the direct action of telomerase. Whereas het-ITSs seem to be intrinsically prone to breakage, the instability of s-ITSs is more controversial. This observation is consistent with the hypothesis that s-ITSs are probably not themselves prone to breakage but represent 'scars' of ancient breakage that may have occurred within fragile regions. This study will review the current knowledge on these two types of ITS, their molecular organization, how they arose during evolution, their implications for chromosomal instability and their potential applications as phylogenetic/forensic markers. PMID- 19188691 TI - Studying telomere replication by Q-CO-FISH: the effect of telomestatin, a potent G-quadruplex ligand. AB - Telomere replication is a critical process for preserving genome integrity. The telomere replication fork proceeds unidirectionally from the last subtelomeric origin towards the end of the chromosome, replicating the 5'-3' G-rich strand by lagging mechanisms and the complementary C-rich strand by leading mechanisms. It has been proposed that the G-rich nature of telomeres may favor the formation of secondary structures such as G-quadruplexes during replication and that specific mechanisms must prevent this to allow the fork to progress unimpeded. The potential of G-quadruplex formation by telomeric sequences has been clearly demonstrated in vitro but it is not known whether these structures form in vivo. We tested the effect of a potent and specific G-quadruplex ligand, telomestatin (TMS), on telomere replication using a novel quantitative approach applied to CO FISH. We show that TMS, although it penetrates and persists within cells, does not affect telomere replication after short or long-term treatments of mouse embryonic fibroblasts. It does however affect the hybridization efficiency of FISH telomeric probes that recognize the G-rich strand. Our work illustrates the use of a novel technique to measure telomere replication efficiency and suggests that G-quadruplex ligands do not affect telomere replication in a non tumoral context. PMID- 19188692 TI - Laser confocal microscopy analysis of human interphase nuclei by three dimensional FISH reveals dynamic perinucleolar clustering of telomeres. AB - Nuclear functions are strongly dependent on the three-dimensional organization of the interphase nucleus. Given the importance of telomeres in the behaviour and stability of chromosomes, we have investigated the architecture of the human nucleus from the telomere perspective by 3D-FISH and laser confocal microscopy. We observed a randomly scattered telomere distribution in all confocal sections of the interphase nuclear volume with various levels of telomere clustering in different cell types. This distribution is independent of H2AX presence or phosphorylation status. We also observed that telomeres usually cluster at the periphery of the nucleolus following its cell cycle dependent dynamic formation but are never present in the interior of the nucleolus. These perinucleolar telomeric clusters contain the telomeres of the short arms of acrocentric chromosomes, explaining the p-arm association of acrocentric chromosomes frequently found in metaphase. Thus, chromosome positioning in metaphase spreads is tightly connected to the three-dimensional architecture of the interphase nucleus. PMID- 19188693 TI - Telomerase recruitment to telomeres. AB - The ability of most cancer cells to grow indefinitely relies on the presence of functional telomerase to maintain telomeres, thus circumventing normal cellular senescence. A key feature of telomerase functionality is the localization of the enzyme complex to telomeres, a process which is highly regulated. A number of recent studies have reported data with significant implications for our understanding of telomerase recruitment to telomeres. A picture is emerging that this process is governed by a number of factors including telomeric binding proteins, structural features of the enzyme complex, cell cycle regulated processes such as DNA replication, and components of the DNA damage response pathway. In this review we summarize recent findings relating to this fundamental process in eukaryotes. PMID- 19188694 TI - Telomerase: cellular immortalization and neoplastic transformation. Multiple functions of a multifaceted complex. AB - The telomerase complex allows telomere length maintenance, which is required for an unlimited cellular proliferation. Telomerase is virtually absent in normal human somatic cells, which are characterized by a definite proliferation potential, while it is present in the vast majority of tumors (around 90%). Restoring telomerase activity in normal somatic cells can indefinitely prolong cellular life span. However, evidence has been reported that this event can be associated with the acquisition of characteristics typical of cellular transformation. Moreover, analysis of telomerase immortalized cells, as well as of tumor cells in which telomerase is inactivated, has highlighted multiple functions of telomerase in tumorigenesis, besides telomere lengthening. In this paper, we will review telomerase immortalization of somatic cells, together with its possible consequences, and we will examine the complex role of telomerase in tumorigenesis. PMID- 19188695 TI - Telomerase regulation at the crossroads of cell fate. AB - Telomeres are specialized structures at the ends of eukaryotic chromosomes and are crucial for genome stability, cell growth control and carcinogenesis. Normally, they protect chromosomes from end to end fusion, degradation and recombination. Telomerase is a ribonucleoprotein essential for maintenance of telomeres and it is active in germ cells, stem cells and approximately 90% of cancers but not in most normal somatic cells. Human telomerase catalytic protein subunit hTERT is crucial for telomerase activity. Although hTERT expression is sufficient to immortalize normal human cells in culture, spontaneous immortalization is extremely rare which suggests that its expression is under strong negative control. Characterization of the hTERT promoter has allowed for the analysis of potential mechanisms of hTERT expression and regulation. The hTERT promoter is very complex and contains a great number of canonical and non canonical sequences that bind or potentially bind a variety of transcription factors. In this review we focus on the positive and negative regulators of hTERT transcription and their role in normal cell growth and immortalization. PMID- 19188696 TI - Dissecting the non-canonical functions of telomerase. AB - It is now well established that the canonical function of telomerase protects the telomere repeats from erosion and the consequent induction of replicative senescence or apoptosis. In the absence of key cell cycle checkpoint proteins, the canonical function of telomerase also prevents chromosome fusions and immortalizes human cells. The canonical function of telomerase requires both the telomerase reverse transcriptase enzyme (TERT) which adds telomere (TTAGGG) repeats to the chromosome ends and the telomerase RNA component (TERC), which provides the template for TERT. However, there is growing evidence that telomerase has other (non-canonical) functions. These functions can be divided further into those that require telomerase activity but not telomere lengthening (non-canonical I or NC I) and those that require neither telomerase activity nor telomere lengthening (non-canonical II or NC II). NC I functions are associated with the induction of neoplasia in both epidermis and mammary gland, the correct response to DNA damage, and insensitivity to transforming growth factor beta. In contrast, NC II functions are not sufficient for the induction of neoplasia and are associated with the activation of the WNT and MYC signaling pathways in keratinocytes and a more general resistance to the induction of apoptosis by a variety of stimuli. The overexpression of either TERT or TERC appears to be capable of providing NC I functions but NC II functions require neither TERC nor the integrity of the TERT catalytic site. The molecular mechanisms underpinning both NC I and NC II are largely obscure but transcriptional profile changes have been reported by some groups. In this article, we will discuss the proposed mechanisms of NC I and NC II and their relevance to normal and neoplastic cell functions. PMID- 19188697 TI - Telomere length maintenance--an ALTernative mechanism. AB - The Alternative Lengthening of Telomeres (ALT) mechanism is utilised by approximately 10% of human tumours and a higher proportion of some types of sarcomas. ALT+ cell lines and tumours show heterogeneous telomere length, extra chromosomal circular and linear telomeric DNA, ALT associated promyelocytic bodies (APBs), a high frequency of post-replication exchanges in telomeres (designated as telomere-sister chromatid exchanges, T-SCE) and high instability at a GC-rich minisatellite, MS32 (D1S8). It is clear that there is a link between the minisatellite instability and the mechanism that underpins ALT, however currently the nature of this relationship is uncertain. Single molecule analysis of telomeric DNA from ALT+ cell lines and tumours has revealed complex telomere mutations that have not been seen in cell lines or tumours that express telomerase. These complex telomere mutations cannot be explained by T-SCE but must arise by another inter-molecular process. The break-induced replication (BIR) model that may explain the observed high frequency of T-SCE and the presence of complex telomere mutations is reviewed. PMID- 19188698 TI - A possible role for telomerase RNA and telomere length in global mitotic recombination. AB - Telomeres are specialised structures at the ends of mammalian chromosomes with many unique properties. Recombinational events at telomeres are more frequent than in the remainder of the genome by several orders of magnitude. This study examined the influence of telomerase status and telomere length on genome-wide recombination assessed by genomic sister chromatid exchange (G-SCE). Telomerase deficiency per se appears to increase G-SCE frequencies in splenocytes but as telomeres shorten through subsequent generations of mTerc(-/-) mice this increase is progressively lost. Telomerase status and telomere length also influences the induction of G-SCE by UV light. Even when mitotic recombination is affected by PARP deficiency, mTerc and telomere length interact to further affect G-SCE frequencies. Taken together the data presented here demonstrate that telomerase status and telomere length can affect recombination frequencies genome-wide. PMID- 19188700 TI - Mechanisms of telomeric instability. AB - Telomeres are the nucleoprotein structures that cap the ends of linear chromosomes. The length of telomeric DNA is an important determinant of function; with short telomeres triggering either replicative senescence or, in the absence of a functional DNA damage response, telomere fusion. Telomere fusion can trigger cycles of anaphase-bridging, breakage and fusion that can lead to genomic arrangements, of the type that are frequently found in cancer. Telomere erosion as a function of ongoing cell division results in the gradual loss of sequences from the terminus; this is superimposed by additional mutational mechanisms that generate large-scale, apparently sporadic, telomeric deletions. These events occur in normal human cells, irrespective of telomerase activity, and because they create telomeres that are capable of fusion, they represent a significant cell-intrinsic mutational mechanism. Here I review some of the potential mechanisms that may result in sporadic telomeric deletion. PMID- 19188701 TI - The number of dysfunctional telomeres in a cell: one amplifies; more than one translocate. AB - Chromosomal instability is increasingly appreciated as a key component of tumorigenesis in humans. A combination of abnormal telomere shortening and cell cycle checkpoint deficiency has been proposed as the initial lesions causing destabilizing chromatin bridges in proliferative cells. We examined the participation of the different types of end-to-end fusions in generating instable karyotypes in non-transformed human breast epithelial cells. We concluded that short dysfunctional telomeres represent an initiating substrate for post replicative rejoining of sister chromatids and are likely to make an important contribution to the formation of chromosomal rearrangements and the amplification of chromosome arm segments in breast epithelial cells. We propose that there is a chronological order in the participation of the different types of end-to-end fusions in the generation of chromosomal instability. Thus, intrachromosomal post replicative joining would proceed mainly in the early stages after overcoming growth arrest, when telomere dysfunction is limited and affects only one chromosome end in a cell. The absence of a second substrate for end joining will conduct the cell with the uncapped chromosome to replicate its DNA and fuse the uncapped sister chromatids after replication. Later, since telomeres shorten progressively with each DNA replication round, the uncapping will affect many more chromosome ends, and fusions between the uncapped ends from different chromosomes will be produced. While the fusion of sister chromatids will produce chromosome segment amplification and terminal deletions in the daughter cells, interchromosomal fusion will produce unbalanced rearrangements other than chromosome segment amplifications. PMID- 19188702 TI - Impaired telomerase activity in human cells expressing GFP-Ku86 fusion proteins. AB - The Ku heterodimer is a DNA end-binding protein that promotes the non-homologous end joining (NHEJ) pathway of DNA double strand break (DSB) repair by recruiting the catalytic subunit of the DNA-dependent protein kinase (DNA-PKcs). Ku is also a normal component of telomeres where it is required for telomere maintenance, interacting not only with the DNA but also with various telomere proteins including telomerase. The way in which Ku simultaneously plays such distinct roles, end-joining at DSBs and end-maintenance at telomeres, is unclear. One way to address this is to study cells in which the NHEJ and telomeric roles of Ku have been separated. Here we describe human cells that express fusions between the large human Ku subunit (Ku86) and a fluorescent protein tag. These cells have reduced telomerase activity and increased sensitivity to ionizing radiation (IR) but no change in their DNA-PK activity or in the DNA end-binding of endogenous Ku. Cells with particularly large amounts of one fusion protein undergo progressive telomere shortening and cellular senescence. These data are consistent with models in which Ku recruits telomerase to telomeres or activates recruited telomerase and suggest that the Ku86 fusion proteins specifically block this role. PMID- 19188699 TI - Telomeres, histone code, and DNA damage response. AB - Genomic stability is maintained by telomeres, the end terminal structures that protect chromosomes from fusion or degradation. Shortening or loss of telomeric repeats or altered telomere chromatin structure is correlated with telomere dysfunction such as chromosome end-to-end associations that could lead to genomic instability and gene amplification. The structure at the end of telomeres is such that its DNA differs from DNA double strand breaks (DSBs) to avoid nonhomologous end-joining (NHEJ), which is accomplished by forming a unique higher order nucleoprotein structure. Telomeres are attached to the nuclear matrix and have a unique chromatin structure. Whether this special structure is maintained by specific chromatin changes is yet to be thoroughly investigated. Chromatin modifications implicated in transcriptional regulation are thought to be the result of a code on the histone proteins (histone code). This code, involving phosphorylation, acetylation, methylation, ubiquitylation, and sumoylation of histones, is believed to regulate chromatin accessibility either by disrupting chromatin contacts or by recruiting non-histone proteins to chromatin. The histone code in which distinct histone tail-protein interactions promote engagement may be the deciding factor for choosing specific DSB repair pathways. Recent evidence suggests that such mechanisms are involved in DNA damage detection and repair. Altered telomere chromatin structure has been linked to defective DNA damage response (DDR), and eukaryotic cells have evolved DDR mechanisms utilizing proficient DNA repair and cell cycle checkpoints in order to maintain genomic stability. Recent studies suggest that chromatin modifying factors play a critical role in the maintenance of genomic stability. This review will summarize the role of DNA damage repair proteins specifically ataxia telangiectasia mutated (ATM) and its effectors and the telomere complex in maintaining genome stability. PMID- 19188703 TI - BRCA1 knock-down causes telomere dysfunction in mammary epithelial cells. AB - A breast cancer predisposing gene, BRCA1, is a major suppressor of chromosomal instability and its dysfunction affects multiple pathways involved in DNA damage response. There is increasing evidence that the mechanisms involved in maintenance of telomeres, specialized structures at chromosome ends, are linked with DNA damage response. We therefore investigated whether BRCA1 dysfunction affects telomere maintenance. To achieve this we knocked-down BRCA1 in two mammary epithelial cell lines using RNA interference. Subsequent analysis by immunofluorescence, RT-PCR and Western blotting revealed that a short interfering RNA oligonucleotide used was able to knock-down BRCA1 efficiently. This knock down did not have any effect on telomerase enzyme activity and telomere length. However, BRCA1 knock-down correlated with the increase in chromatin bridges in anaphase cells which usually reflect telomere dysfunction. Therefore, this study suggests that BRCA1 knockdown in mammary epithelial cells causes telomere dysfunction. PMID- 19188704 TI - Increased genomic alteration complexity and telomere shortening in B-CLL cells resistant to radiation-induced apoptosis. AB - B-cell chronic lymphocytic leukemia (B-CLL) results in an accumulation of mature CD5(+)/CD23(+) B cells due to an uncharacterized defect in apoptotic cell death. B-CLL is not characterized by a unique recurrent genomic alteration but rather by genomic instability giving rise frequently to several chromosomal aberrations. Besides we reported that approximately 15% of B-CLL patients present malignant B cells resistant to irradiation-induced apoptosis, contrary to approximately 85% of patients and normal human lymphocytes. Telomere length shortening is observed in radioresistant B-CLL cells. Using fluorescence in situ hybridization (FISH) and multicolour FISH, we tested whether specific chromosomal aberrations might be associated with the radioresistance of a subset of B-CLL cells and whether they are correlated with telomere shortening. In a cohort of 30 B-CLL patients, all of the radioresistant B-CLL cell samples exhibited homozygous or heterozygous deletion of 13q14.3 in contrast to 52% of the radiosensitive samples. In addition to the 13q14.3 deletion, ten out of the 11 radioresistant B-cell samples had another clonal genomic alteration such as trisomy 12, deletion 17p13.1, mutation of the p53 gene or translocations in contrast to only three out of 19 radiosensitive samples. Telomere fusions and non-reciprocal translocations, hallmarks of telomere dysfunction, are not increased in radioresistant B-CLL cells. These findings suggest (i) that the 13q14.3 deletion accompanied by another chromosomal aberration is associated with radioresistance of B-CLL cells and (ii) that telomere shortening is not causative of increased clonal chromosomal aberrations in radioresistant B-CLL cells. PMID- 19188705 TI - Genomic imbalances in key ion channel genes and telomere shortening in sudden cardiac death victims. AB - Sudden cardiac death (SCD) can be caused by a number of reasons. Previous works have identified the genetic causes, such as alterations in the DNA sequence, for many of these diseases. We hypothesize that some patients may show genomic imbalances and changes in the gene copy number leading to genetic instability. To clarify this, we analysed DNA samples from SCD victims using comparative genomic hybridization (CGH), a molecular cytogenetic technique that permits the genome wide screening of chromosomal imbalances, and telomere length measurement. DNA derived from peripheral blood and heart tissue of 14 SCD cases and six apparently healthy control individuals were subjected to CGH analysis. Telomere length measurements were done by the Southern blotting method. Eight out of 14 SCD cases exhibited changes in DNA/gene copy number. CGH analysis showed variation in the gene copy number of some of the genes associated with potassium (KCNAB1, KCNH2, and KCNA4) and calcium (RyR2, ATP2A2) ions which are involved in maintaining the ionic balance of the heart. Alterations in TERC and TERT genes were also detected in SCD victims. In nine SCD victims shorter telomeres were detected. This might have resulted from excessive cellular proliferation and/or oxidative stress in these individuals. Copy number changes observed and telomere shortening detected in SCD cases would possibly explain at least some of the causes of SCD at early ages in humans. Identification of biomarkers of SCD is of great importance and thus the present study will facilitate the identification of some of the biomarkers. PMID- 19188707 TI - Telomere capping and cellular checkpoints: clues from fruit flies. AB - In most organisms, telomeres consist of repetitive G-rich sequences that are elongated by a specific reverse transcriptase, telomerase. A large number of proteins are recruited by these terminal repeats, forming specialized structures that regulate telomerase activity and protect telomeres from degradation and recombination. Drosophila lacks telomerase and telomere length is maintained by transposition of three specialized retrotransposons. In addition, unlike yeast and mammals, Drosophila telomeres are epigenetically determined, sequence independent structures. However, several proteins required for Drosophila telomere behavior are evolutionarily conserved. These include the Mre11-Rad50-Nbs (MRN) complex and the Ataxia Telangiectasia Mutated (ATM) kinase, which are required to prevent telomeric fusions. In addition, recent studies have provided evidence that Drosophila uncapped telomeres elicit a DNA damage response (DDR) just as dysfunctional yeast and human telomeres. Uncapped Drosophila telomeres also activate the spindle assembly checkpoint (SAC) by recruiting the SAC kinase BubR1. Telomere-induced DDR and SAC both require the wild type function of the MRN complex. In addition, while DDR is mediated by ATR kinase, SAC activation requires both the ATM and ATR activities. These results indicate that the DNA repair systems play multiple roles at Drosophila telomeres, highlighting the importance of this model organism for investigations on the relationships between DNA repair and telomere maintenance. PMID- 19188706 TI - Regulation of telomere length in Drosophila. AB - Telomeres in all organisms must perform the same vital functions to ensure cell viability: to act as a protective chromosome cap that distinguishes natural chromosome ends from DNA double strand breaks, and to balance the loss of DNA from the chromosome end due to incomplete DNA replication. Most eukaryotes rely on a specialized reverse transcriptase, telomerase, to generate short repeats at the chromosome end to maintain chromosome length. Drosophila, however, uses retrotransposons that target telomeres. Transposition of these elements may be controlled by small RNAs and spreading of silent chromatin from the telomere associated sequence, both of which limit the retrotransposon expression level. Proteins binding to the retrotransposon array, such as HP1 and PROD, may also modulate transcription. It is not clear however, that simply increasing transcript levels of the telomeric retrotransposons is sufficient to increase transposition. The chromosome cap may control the ability of the telomere specific elements to attach to chromosome ends. As in other organisms, chromosomes can be elongated by gene conversion. Although the mechanism is not known, HP1, a component of the cap, and the Ku proteins are key components in this pathway. PMID- 19188708 TI - Cytogenetics for the study of telomere function in plants. AB - The ends of linear chromosomes of the vast majority of eukaryotic species possess specialized nucleo-protein structures called telomeres. Regardless of many exceptions, the structure and function of telomeres share high degrees of similarity between various eukaryotes. The underlying DNA structure of telomeres determines the particular setup of telomere chromatin and protein complexes as are telomere-associated proteins and a number of repair and cell cycle checkpoint agents. The structure of telomeres is highly dynamic during the cell's growth, replication, differentiation, senescence, or neoplastic transformation. Although the bulk of our knowledge about telomere function comes from molecular and biochemical studies in model organisms such as yeast and mouse, we want to show- with special emphasis on plants--in this short review that classical methods of plant cytogenetics can significantly complement the above experimental approaches and help in our understanding of telomere functions. PMID- 19188709 TI - Two faces of Solanaceae telomeres: a comparison between Nicotiana and Cestrum telomeres and telomere-binding proteins. AB - While most Solanaceae genera (e.g.Solanum, Nicotiana) possess Arabidopsis-type telomeres of (TTTAGGG)n maintained by telomerase, the genera Cestrum, Vestia and Sessea (Cestrum group) lack these telomeres. Here we show that in the Cestrum group the activity of telomerase has been lost. Nevertheless, proteins binding the single-stranded G-rich strand of the Arabidopsis-type and related human-type (TTAGGG)n telomeric sequences are present in nuclear extracts of both Nicotiana and Cestrum species. These proteins may have a role in telomere function or other cellular activities. In addition to characterizing DNA binding specificity and molecular weights of these proteins, we searched in both N. tabacum (tobacco) and C. parqui for the presence of POT1-like proteins, involved in telomere capping and telomerase regulation. Analysis of POT1-like proteins available on public databases and cloned by us from C. parqui, revealed the N-terminal OB folds typical for this protein family and a novel, plant-specific conserved C-terminal OB-fold domain (CTOB). We propose that CTOB is involved in protein-protein interactions. PMID- 19188710 TI - Genome stability in Arabidopsis cells exhibiting alternative lengthening of telomeres. AB - Indefinite proliferation of eukaryotic cells depends on telomerase that counteracts the depletion of DNA from chromosome termini due to the end replication problem. The requirement for telomerase can, under certain conditions, be circumvented by employing homologous recombination-based mechanisms for telomere maintenance. Whereas yeast and mammalian cells lacking telomerase appear to readily adopt alternative telomere lengthening (ALT), in Arabidopsis ALT is inhibited by the Ku heterodimer. Although we failed to establish ALT cell lines from Ku-proficient lines, ALT can be efficiently induced in cells derived from mutants deficient for telomerase reverse transcriptase (TERT) and Ku70. In this study, we describe the growth performance, genome stability and long-term maintenance of telomeric DNA in Arabidopsis ku70 tert ALTcultures. ALT activation increases karyotype stability in the majority of tert ku70 cell lines, which contrasts with ongoing chromosomal rearrangements detected in survival tert cultures that lack any detectable telomeric sequences. Curiously, ku70 tert ALT lines and tert survivor cultures proliferate at a similar rate, although the latter display remarkable chromosomal abnormalities, including giant circular and linear megachromosomes that seem to arise by fusions of multiple chromosomes. This observation underlies the extraordinary tolerance of plant cells to telomere dysfunction. PMID- 19188711 TI - Distribution of (TTAGGG)n telomeric sequences in karyotypes of the Xenopus species complex. AB - The chromosomal distribution of the conserved vertebrate telomeric (TTAGGG)(n) sequence was studied by fluorescence in situ hybridization (FISH) in four Xenopus species and the triploid Silurana tropicalis. As expected, hybridization signals were observed at the distal ends of every chromosome in all species. In addition, the hybridization pattern demonstrates varied organization of (TTAGGG)(n) sequences in the different karyotypes. In X. borealis and X. muelleri hybridization signals intensely labeled one end of a homologous chromosome pair that coincides with the sites containing ribosomal RNA gene clusters. The karyotype of X. clivii remarkably differs from other Xenopus karyotypes in displaying numerous interstitial telomeric sites (ITS). C-banding analysis shows that the non-telomeric sites appear to correspond to the interstitially located constitutive heterochromatin. This suggests that interstitial telomeric sites in X. clivii do not necessarily represent the relic of ancestral telomeres resulting from the fusion of chromosomes, but their occurrence is due to the fact that (TTAGGG)(n) repeat arrays may be a constituent of highly repetitive DNA. PMID- 19188712 TI - Antimicrobial susceptibility pattern of bacterial pathogens causing urinary tract infections in a Saudi Arabian hospital. AB - The study describes the pathogens causing urinary tract infections (UTI) and the antimicrobial susceptibility pattern of health care-associated (HCA) and community-acquired (CA) isolates. Of the total of 3,989 nonrepetitive urinary cultures, 4.7% (n=188) were HCA and 95.3% (n=3,801) were CA isolates. Gram negative organisms totaled 3,607 (90.4%) vs. 9.6% Gram-positive organisms. In CA UTI, Escherichia coli, Klebsiella pneumoniae and Enterococcus faecalis constituted 66, 11.4, and 5.4%, respectively. HCA-UTI episodes were caused by E. coli (37.8%), K. pneumoniae (14.4%) and Pseudomonas aeruginosa (6.4%). The susceptibility rates of E. coli to ciprofloxacin, trimethoprim-sulfamethoxazole (TMP-SMX), nitrofurantoin and ceftriaxone in HCA and CA isolates were 53.3 and 74.1%, 42.2 and 62.7%, 32.5 and 97.0%, and 62.3 and 92.7%, respectively. Extended spectrum beta-lactamase production was detected in 8.1 and 7.4% isolates, respectively, of CA and HCA isolates of E. coli. CA isolates of K. pneumoniae were more susceptible to TMP-SMX (89.2 vs. 60.4%), ciprofloxacin (92.6 vs. 67.9%) and cefuroxime (93.8 vs. 24.5%) than HCA isolates. The susceptibility of HCA and CA isolates of E. faecalis to penicillin G and nitrofurantoin were 34.8 and 80%, and 78.3 and 93.6%, respectively (p<0.001). In conclusion, antimicrobial resistance is high to commonly used oral agents, rendering them inappropriate for empirical use. PMID- 19188713 TI - Activity of nilotinib (AMN-107) alone in advanced gastrointestinal stromal tumors progressing on imatinib and sunitinib. Case report. AB - Nilotinib is a novel tyrosine kinase inhibitor targeting KIT (CD117), PDGFR and BCR-ABL and inhibiting the proliferation of both imatinib-sensitive and imatinib resistant cells in vitro. Nilotinib, alone or in combination with imatinib, has promising activity in imatinib-resistant patients with advanced gastrointestinal stromal tumors (GISTs), including those who progressed on sunitinib and other tyrosine kinase inhibitors. We describe the beneficial effect of nilotinib 400 mg b.i.d. orally in a 53-year-old patient with metastatic GISTs who had radiologically confirmed disease progression on both imatinib (800 mg/day) and sunitinib (50 mg/day). A positron emission tomography-computerized tomography evaluation showed marked decreases in (18)F-fluorodeoxyglucose uptake in the liver and mesentery region after 3 months of therapy with nilotinib. Nilotinib is an option for patients with advanced GISTs progressing on both imatinib and sunitinib. PMID- 19188714 TI - Innovations in interventional endoscopy. Introduction. PMID- 19188715 TI - Endoscopic ablation of Barrett's esophagus using the Halo system. AB - There is increasing interest in the endoscopic treatment of Barrett's esophagus. Endoscopic treatment has been utilized for many years, but in the past, no specific method has emerged as an appealing treatment option with appropriate safety, efficacy and ease of treatment for both patients and physicians. Recently there has been a growing literature related to the endoscopic ablation of Barrett's esophagus using radiofrequency ablation (RFA) (Halo system). In order to discuss when RFA is indicated for Barrett's, one needs to know: (1) What is the 'histology' of the Barrett's? Does the patient have intestinal metaplasia, low-grade dysplasia, high-grade dysplasia or intramucosal carcinoma? (2) What are the endoscopic options to be considered as opposed to RFA? What are the advantages and disadvantages of each? (3) What additional variables need to be examined? PMID- 19188716 TI - Endoscopic resection of early esophageal and gastric neoplasias. AB - The advent of endoscopic resection (ER) techniques has enabled gastroenterologists to remove premalignant or neoplastic lesions throughout the gastrointestinal tract. This review discusses the indications and the several techniques of ER in early carcinomas of the esophagus and stomach. Before ER is performed an accurate evaluation of patients and careful staging of lesions is mandatory. After ER of the neoplasia histological assessment of the entire specimen with detailed histological analysis of layer infiltration is crucial. First long-term follow-up studies of large numbers of patients confirm the excellent effectiveness of ER for well-differentiated mucosal lesions without lymphangitic invasions. PMID- 19188717 TI - Modern endoscopic therapy of peptic ulcer bleeding. AB - Acute upper gastrointestinal bleed (UGIB) remains a challenging clinical problem owing to significant patient morbidity and costs involved with management. Peptic ulcer bleeding (PUB) contributes to the majority of causes of UGIB with a growing concern of its impact on the elderly and the increasing use of non-steroidal anti inflammatory drugs as precipitating bleeding episodes. Apart from initial critical care, endoscopy is the preferred first-line management of PUB. Early use of empirical high-dose proton pump inhibitor therapy prior, during and after endoscopy is cost-effective and reduces the need for endotherapy. Current endoscopic modalities, both thermal and non-thermal, offer a wide range of choices in high-risk PUB (active arterial bleeding or non-bleeding visible vessel). Combinations of injection (epinephrine) along with thermal therapy or endoclips are recommended for better clinical outcomes. The role of endotherapy for adherent clots is controversial. A second-look endoscopy may be beneficial in high-risk patients. A multidisciplinary team approach should be part of all treatment protocols for the ideal management of UGIB. PMID- 19188718 TI - Efficacy, complications and clinical outcomes of endoscopic sclerotherapy with N butyl-2-cyanoacrylate for bleeding gastric varices. AB - BACKGROUND: Bleeding gastric varices (BGV) are a severe complication of portal hypertension. Although the obliteration of gastric varices using cyanoacrylate has been shown effective to eradicate gastric varices, there are still few data available on its effectiveness for acutely BGV, cyanoacrylate-related complications and follow-up. AIMS: To report on the feasibility, efficacy and outcomes of N-buytl-2-cyanoacrylate sclerotherapy in patients with BGV. METHODS: This is a retrospective cohort study of consecutive patients who were evaluated for BGV at University of Magdeburg Medical Center. Data abstracted for analysis from the prospectively collected database included patient's demographics, etiology of gastric varices, Child-Pugh score, effectiveness of endoscopic treatment, clinical and endoscopic follow-up. Immediate hemostasis was defined as visible and clinical arrest of bleeding. The following were considered as potential complications of the procedure: aspiration, fever, worsening hemorrhage due to the sclerotherapy and systemic embolization of cyanoacrylate. RESULTS: 33 patients (17 F, 16 M; mean age 54 years, range 18-81) with BGV (29 had active bleeding and 4 had stigmata of recent bleeding) underwent endoscopic injections of N-butyl-2-cyanoacrylate for hemostasis over a 5-year period. Mean follow-up: 9 months (range: 1 month to 5 years). Initial hemostasis was achieved in 88%. Treatment-related complications occurred in 5 (15.1%), the most common being deep ulcerations, which also resulted in recurrent bleeding. One patient presented with distal cyanoacrylate embolization. During long-term follow-up, 6 patients died, 5 as a consequence of the underlying liver disease and 1 as a consequence of complications due to systemic cyanoacrylate embolization. CONCLUSIONS: N-butyl 2-cyanoacrylate sclerotherapy is highly effective for the treatment of BGV, with rare complications occurring both acutely and in the long term. PMID- 19188719 TI - Laparoscopic fundoplication for GERD: are we there yet? AB - Although proton pump inhibitor treatment remains the first-line treatment for gastroesophageal reflux disease (GERD), the panorama of an indefinitely prolonged medical treatment faces patients and physicians with the need to look for an equivalent, more definitive therapy. Facing global results, after many series have demonstrated the safety of the laparoscopic antireflux surgery procedure in experienced hands, with a low incidence of intra- and postoperative morbidity and a short postoperative hospital stay (averaging 2 days), the immediate benefit as well as the long-term alleviation of symptoms (very good: 82%, good: 92%) seem to justify the place of surgical fundoplication in the therapeutic armamentarium for GERD as a valid alternative to medical treatment. Careful patient selection for surgery, a complete and thorough preoperative workup, an intervention in experienced hands, and a patient well informed about the pros and cons of both medical and surgical therapy are mandatory. PMID- 19188720 TI - Therapeutic balloon-assisted enteroscopy. AB - Since the introduction of the first balloon-based enteroscopic technique in 2001, therapeutic balloon-assisted enteroscopy (BAE) using either the single or double balloon enteroscopy technique (respectively SBE and DBE) has evolved rapidly. Argon plasma coagulation (APC), polypectomy, dilation therapy of strictures, and therapy of the pancreatico-biliary system in patients with surgical altered proximal intestinal anatomy: all have been successfully introduced to treat pathological findings in all segments of the small bowel. The clinical impact of treatment of vascular malformations, strictures caused by chronic inflammation (especially Crohn's disease) and polypectomy therapy (especially in the Peutz Jeghers syndrome) seems evident. The decrease of, often repeated, surgical therapy after successful therapeutic BAE in the latter 2 patient groups appears to be a big step forward in treatment. The development of newer enteroscopes, specialized equipment and improved sedation of patients adds positively to the clinical management of undergoing therapeutic BAE. The overall complication rate of therapeutic BAE seems acceptable, but is higher compared to therapeutic colonoscopy which needs further attention in future. PMID- 19188721 TI - Endoscopic balloon dilation of anastomotic strictures occurring after laparoscopic gastric bypass for morbid obesity. AB - BACKGROUND: Gastrojejunal anastomotic stenosis of laparoscopic Roux-en-Y gastric bypass (LRYGB) for morbid obesity occurs in 3-25% of cases. The aim of this report was to evaluate the utility of endoscopic balloon dilation for the therapy of anastomotic strictures after LRYGB. PATIENTS AND METHODS: 111 consecutive patients were treated with endoscopic dilation under sedation with propofol. Dilations were performed with through-the-scope over-the-wire balloons, with sizes ranging from 6 to 18 mm. The outcomes of the procedure were analyzed. RESULTS: 200 endoscopic balloon dilations were performed in 111 patients. Repeated dilations were necessary in patients with complex stenosis. In 75% of the patients it was possible to dilate to 12 mm during the first session. Only in 2% of the cases was it impossible to introduce the endoscope through the stenosed anastomosis after dilation. On follow-up a repeat dilation was necessary in 26% of the cases. Minor complications occurred in 2.7% of patients (2 concealed perforations and 1 hematoma of the esophagus). These were treated conservatively. None of the patients required operation. CONCLUSIONS: Endoscopic balloon dilation is a safe and effective therapy for anastomotic strictures occurring after LRYGB. PMID- 19188723 TI - ERCP with the balloon-assisted enteroscopy technique: a systematic review. AB - BACKGROUND: Endoscopic retrograde cholangiopancreatography (ERCP) is generally difficult with conventional instruments in patients with surgically altered gastrointestinal anatomy. Balloon-assisted enteroscopy is a relatively new technique that allows for access to the papilla of Vater or a biliodigestive anastomosis in postsurgical reconstructions. AIM: To systematically review the utility of balloon-enteroscope-assisted ERCP for patients with altered gastrointestinal anatomy. MATERIAL AND METHODS: Systematic search of the literature on diagnostic and therapeutic interventions in the pancreaticobiliary system using balloon-assisted enteroscopy in patients with previous intestinal surgery indexed in of MEDLINE, Current Contents, PubMed, and references from relevant articles using the search terms 'balloon', 'enteroscopy', 'endoscopy', 'ERCP' and 'cholangiography'. Only papers published as 'full text' in English were included. RESULTS: In total, 16 relevant reports are available on balloon assisted procedures performed in 63 patients with a variety of anatomical configurations. Double balloon procedures as well as single balloon techniques have been reported. Successful interventions in these patients include balloon dilation of stenotic biliodigestive anastomoses, sphincterotomy, extraction of bile stones and stent placement. No serious complications have yet been reported. CONCLUSIONS: Balloon-assisted enteroscopy is a safe and feasible technique to obtain biliary or pancreatic access in patients with surgically altered anatomical configurations. The diagnostic and therapeutic potential of balloon assisted ERCP is large, and could be further improved if customized accessories become more widely available. PMID- 19188722 TI - Small bowel endoscopy using the double-balloon technique: four-year results in a tertiary referral hospital in Brazil. AB - BACKGROUND: Double-balloon enteroscopy (DBE) allows evaluation and therapy for various small bowel diseases. In this series the outcome of a 4-year experience in a tertiary hospital school in Brazil is reported. METHODS: A total of 457 consecutive DBE were performed in 418 patients from August 2004 to August 2008. 93 patients with several indications, whose aim was not the evaluation of suspected diseases of the small bowel mucosa, were excluded, therefore leaving 364 DBE in 325 patients for analysis. Data were retrospectively collected with regard to clinical, endoscopic findings, therapy and complications. RESULTS: Among the 364 DBE performed in 325 patients, 143/325 were males (44%) and 182/325 females (56%) with a mean age of 48.6 +/- 15.7 years (range 17-89). Mean investigation time was 64 +/- 22 min (range 35-135). The depth of insertion beyond the ligament of Treitz was 230 +/- 85 cm (range 30-500) by the antegrade approach and 140 +/- 75 cm (range 0-320) by the retrograde approach. Total enteroscopy was achieved in 41.66% of the attempts (30 of 72 patients). Overall diagnostic yield was 54.95% (200 of 364 procedures) ranging from 0 to 100% in this series, depending on the indication. Angiodysplasia was the main diagnosis in 24.5% (49 of 200 procedures) and endoscopic treatment, including biopsies, hemostasis, tattooing and polypectomy were performed in 65.38% (238 of 364 procedures). No major complications were reported. CONCLUSIONS: DBE is a feasible, safe and well-tolerated procedure allowing endoscopic therapy. Selection of indications increases its diagnostic yield. PMID- 19188724 TI - ERCP using double-balloon enteroscopy in patients with Roux-en-Y anatomy. AB - Double-balloon enteroscopy (DBE) is a useful method for endoscopic retrograde cholangiopancreatography (ERCP) in patients with Roux-en-Y anatomy. Depending on the distorted anatomy, endoscopic therapies with conventional scopes were very difficult or impossible before the advent of DBE and patients had to be submitted to a percutaneous or surgical approach. The case of 6 patients with different types of Roux-en-Y-altered anatomy in which DBE-ERCP was performed with 83.3% successful rate (5/6) is reported confirming recent data in the literature on the feasibility of this method. PMID- 19188725 TI - Colonic stents for the palliation of malignant colonic obstruction. AB - Colorectal self-expanding metal stents have revolutionized the palliative treatment of patients with advanced malignancies and those patients who present with acute colon obstruction and who were previously treated with emergent surgical decompression via formation of colostomy. The placement of stents can be fluoroscopically or non-fluoroscopically guided. The use of stents is associated with significant reduction in hospital duration, mortality and medical complications compared to surgery. The most frequent complications are stent malposition and migration, bleeding, colonic perforation and tumor ingrowth and/or overgrowth. PMID- 19188726 TI - Polypectomy techniques for difficult colon polyps. AB - A difficult polyp is any flat or raised colonic mucosal lesion that given its size, shape or location makes it difficult for the colonoscopist to remove. Although many 'difficult polyps' will be an easy target for the advanced endoscopist, polyps that are >15 mm, have a large pedicle, are flat and extended, are difficult to see or are located in the cecum or any angulated portion of the colon should be always considered difficult. Post-polypectomy complications are more common in the presence of difficult polyps. This review describes several useful tips and tricks to deal with such polyps. PMID- 19188727 TI - Interventional endoscopic ultrasound. AB - Endoscopic ultrasonography (EUS), since its development in the 1980s, has undergone a great deal of technological advancement in imaging, scopes with larger channels and accessories. This has given the endosonographer an immense reach in terms of accessing difficult areas with fewer complications. Currently, it is possible not only to obtain tissue for histological diagnosis but also to perform therapeutic procedures that hitherto would have needed surgery with its attendant morbidity. EUS has evolved over the years and EUS-guided fine needle aspiration for histological diagnosis has become standard practice. A wide array of interventional procedures are performed under EUS guidance including, celiac plexus neurolysis, drainage of pancreatic and pelvic fluid collections, drainage of obstructive biliary/pancreatic ducts, and implantation of fiducial markers/radioactive seeds into gastrointestinal tumors. Also, various experimental procedures are underway evaluating the role of EUS in radiofrequency ablation, vascular therapy, and natural orifice transluminal endoscopic surgery. In this review, we examine the various EUS-guided interventions currently undertaken and explore the potential of other emerging experimental techniques. PMID- 19188728 TI - Prospective assessment of diagnostic utility and complications of endoscopic ultrasound-guided fine needle aspiration. Results from a newly developed academic endoscopic ultrasound program. AB - BACKGROUND: It is currently unknown whether a newly developed endoscopic ultrasound (EUS) program can produce results similar to those of experienced EUS centers. We therefore prospectively evaluated the diagnostic accuracy and major complications of EUS-guided fine needle aspiration (EUS-FNA) in a newly developed EUS program. METHODS: All procedures were performed by a single endosonographer in the presence of a cytopathologist. Reference standard for classification of final disease included: surgical resection, death from disease progression and repeat radiologic and/or clinical follow-up. Major complications were defined as oversedation, and those that resulted in a physician or emergency department visits, hospitalization, or death. RESULTS: 540 patients (median age 63 years, 77% white) underwent EUS-FNAs of 656 lesions: lymph nodes (LNs, n = 248), solid pancreatic masses (SPMs, n = 229), cystic pancreatic masses (CPM, n = 57), mural lesions (n = 41), bile duct/gallbladder (n = 28), liver (n = 17), mediastinum/lung (n = 17), adrenal (n = 15), spleen (n = 3) and kidney (n = 1). SPMs and bile duct/gallbladder lesions were more likely to have suspicious/atypical cytology when compared to other lesions (8.7 vs. 4.6%; p = 0.04) and required more passes to achieve a tissue diagnosis (p < 0.001). The overall sensitivity, specificity, PPV, NPV and accuracy of EUS-FNA was 91.7, 97.1, 98.1, 87.7 and 93.8%, respectively. Six patients (1.1%) (95% CI 0.4-2.4) experienced a major complication. One patient died shortly after the procedure due to preexisting pulmonary embolus (0.18%). CONCLUSION: EUS-FNA is highly accurate and safe in sampling a variety of peri-intestinal organs and lymph nodes. With adequate third-tier training, a newly developed program can produce results similar to those from expert EUS centers. PMID- 19188729 TI - Management of infected and sterile pancreatic necrosis by programmed endoscopic necrosectomy. AB - INTRODUCTION: Necrosectomy is the gold standard treatment for infected pancreatic necrosis (IPN). A percutaneous and endoscopic approach has been accepted in selected cases. Endoscopic drainage (ED) of IPN can be performed by using transpapillary or transmural procedures, or a combination of both with or without endoscopic ultrasound. AIMS: The aim of this study was to determine the indications, complications, success rate, and the importance of assessment of main pancreatic duct integrity by endoscopic retrograde pancreatography (ERP) in patients with IPN. METHODS: Records of all patients who underwent endoscopic necrosectomy from January 2002 to December 2007 at Rio de Janeiro Federal University Hospital were reviewed. A total of 56 patients were included. ED was performed using daily transmural and transpapillary drainage. A diagnostic pancreatogram (ERP) to search for communications between the pancreatic duct and the collection were performed in all cases and in cases where communication existed. A pre-cut needle knife was used to puncture the cyst wall, aspirate the content and then enter at the cyst cavity (contrast was injected to ensure opacification of the cyst and subsequent drainage). Sphincterotomy catheter or balloons were used to enlarge and ensure a wide cystoenterostomy. All patients were followed with computerized tomography scans or ultrasound to ensure clinical resolution. Mean follow-up was 21 months. RESULTS: 49/56 patients could be successfully treated. ED was successful in 49 patients (87%) and in 3 (13%) it failed. Mean follow-up was 21 months. During this period, there were 2 (10.5%) pseudocyst recurrences and only 1 (5.2%) recurrence of new episodes of pancreatic necrosis, and all were managed clinically and/or endoscopically. No mortality was related to the procedure. CONCLUSION: ED with daily necrosectomy is a useful method to remove infected and sterile pancreatic necrosis. PMID- 19188730 TI - Single-step EUS-guided endoscopic treatment for sterile pancreatic collections: a single-center experience. AB - BACKGROUND AND AIMS: Endoscopic ultrasound (EUS) is useful for the treatment of sterile pancreatic fluid collections (PFC), either by means of transmural drainage or by complete aspiration. The aim of this study was to evaluate the efficacy and safety of single-step EUS-guided endoscopic approaches for treatment of sterile PFC. PATIENTS AND METHODS: During a 3-year period, 77 consecutive patients with symptomatic, persistent sterile PFC were evaluated and treated with the linear EUS. We excluded patients with grossly purulent collections, chronic pseudocyst and those whose cytology diagnostic was neoplastic cyst of pancreas. 44 patients received a single 10-Fr plastic straight stent under EUS or fluoroscopic control (group I) and 33 of these underwent a single-step complete aspiration with a 19-gauge needle (group II). RESULTS: The mean size of the sterile PFC was 48 mm in group I and 28 mm in group II (p < 0.001). Overall, endoscopic treatment was successful in 70 (90.9%) patients. The mean volume aspirated was 25 (18-65) ml. The total number of procedures was 50 in group I and 41 punctures in group II. After a mean follow-up of 64 +/- 15.6 weeks there were 6 complications (13.6%): 2 recurrences (referred to surgery), 2 developing abscesses (submitted a new EUS-guided endoscopic drainage with success), 1 perforation that died (2.2%), and 1 case of bleeding (sent to surgery) in group I. In group II there were only 6 (18.1%) recurrences (submitted a new EUS-guided aspiration). None of the patients undergoing single-step aspiration developed infections, perforation or hemorrhage. CONCLUSION: The recurrence of pancreatic pseudocysts after endoscopic treatment was similar, either by means of plastic stents or by complete single-step aspiration. PMID- 19188731 TI - Role of repeated endoscopic ultrasound-guided fine needle aspiration in small solid pancreatic masses with previous indeterminate and negative cytological findings. AB - INTRODUCTION: Despite advances in imaging techniques, the differentiation between pancreatic cancer and benign lesions remains difficult. Endoscopic ultrasound guided fine needle aspiration (EUS-FNA) is an effective method for providing tissue diagnosis, but problems occur when lesions are small or the cytological diagnosis is indeterminate. AIM: To prospectively evaluate the utility of EUS-FNA in patients with small solid pancreatic lesions and those with initial indeterminate or negative cytological diagnosis. METHODS: During the study period we performed a total of 119 EUS-FNA procedures on 46 patients (mean age 56.3 years) for 47 small solid pancreatic lesions (range 7-30 mm, mean 17.2 mm in diameter). FNAs were performed in the presence of a cytopathologist. If cytological diagnoses were indeterminate, EUS-FNA was repeated within 3 weeks. Diagnoses were confirmed histologically or by follow-up (clinical and imaging: EUS +/- FNA and CT). RESULTS: Localization of the lesions: head 28 (60%), uncinate process 4 (9%), body 11 (23%) and tail 4 (9%). On average, 3.7 passes were performed. We observed no complications. Initial cytological findings were: malignant 17 (36%), benign 21 (45%), and indeterminate 9 (19%). 8 (78%) of the indeterminate findings were confirmed to be malignant on repeated procedures. A diagnosis of pancreatic cancer was subsequently confirmed in 1 patient who had a benign cytological finding. 19 patients underwent surgery. Histology confirmed a neoplasm in all cases. Sensitivity, specificity, positive predictive value, negative predictive value and diagnostic accuracy were 68, 100, 100, 73 and 83%, respectively. After repeated EUS-FNAs of indeterminate findings sensitivity, negative predictive value and diagnostic accuracy rose to 92, 77 and 96%, respectively. CONCLUSION: EUS-FNA is a highly effective method for providing tissue diagnosis in patients with small solid pancreatic masses. Repeated procedures enhanced diagnostic accuracy in indeterminate findings, among which was high percentage of malignancies. EUS-FNA reduced the number of operations in patients with pancreatic solid masses. PMID- 19188732 TI - Secondary lymphangiectasia of the small bowel: utility of double balloon enteroscopy for diagnosis and management. AB - Sporadic lymphangiectasias are commonly found throughout the small bowel and are considered to be normal. Not uncommonly, lymphangiectasias are pathologic and can lead to mid-gastrointestinal bleeding, abdominal pain and protein-losing enteropathy. Pathologic lymphangiectasias of the small bowel include primary lymphangiectasia, secondary lymphangiectasia and lymphaticovenous malformations. In this report we present three different cases of small bowel lymphangiectasia detected by double balloon enteroscopy. The patients were diagnosed with South American blastomycosis, tuberculosis and primary small bowel lymphangioma. PMID- 19188733 TI - An unusual cause of abdominal pain. PMID- 19188734 TI - A 'double-balloon enteroscopy worth the money': endoscopic removal of a coin lodged in the small bowel. AB - Double-balloon enteroscopy (DBE) has become the standard endoscopic method to diagnose and treat disorders of the small bowel. The most common therapeutic applications of DBE are hemostasis, polypectomy and stricture dilation. DBE has also been used to place stents in the small bowel and to retrieve foreign bodies such as retained capsule endoscopes. In this report, we describe the removal of a coin that had remained lodged in the jejunum for 8 days. PMID- 19188735 TI - Diagnostic and therapeutic applications of endoscopic ultrasound-guided punctures. AB - Endoscopic ultrasound (EUS) has emerged as an excellent tool for the imaging of the gastrointestinal wall and surrounding structures. EUS-guided fine needle aspiration has broadened the applicability of this tool by allowing tissue sampling of a variety of lesions within or accessible from the gastrointestinal (GI) tract. In particular, EUS became the test of choice for evaluating pancreatic cysts and mass lesions, biliary strictures and masses, abnormal adenopathy accessible from the GI tract, and GI submucosal lesions. There appears to be an increasing role for EUS in the staging of lung cancer, evaluating adrenal lesions and GI mesenchymal tumors. More recently, EUS has acquired a new dimension in interventional applications. This includes, but is not limited to, celiac plexus block and celiac neurolysis for the management of pain associated with chronic pancreatitis and locally advanced pancreaticobiliary malignancies, respectively. PMID- 19188736 TI - Valproate-induced insulin resistance and obesity in children. AB - BACKGROUND: Valproic acid (VPA), a widely used antiepileptic drug, has broad spectrum activity against both generalized and partial epilepsy. Among the side effects of VPA, weight gain is frequently reported, although the real incidence and magnitude of this problem is unknown. Its pathogenesis is most likely multifactorial, and is controversial. METHODS: In order to evaluate the role of hyperinsulinemia and related hormonal abnormalities in VPA-induced obesity, data from the existing literature have been analyzed and discussed critically. RESULTS: Patients suffering from weight gain show various metabolic and endocrinologic abnormalities. The most frequent are hyperinsulinemia and insulin resistance, hyperleptinemia and leptin resistance, and an increase in the availability of long-chain free fatty acids. Significant weight gain is associated with increased levels of insulin and leptin, suggesting a close relationship between obesity-induced hyperinsulinemia and hyperleptinemia. VPA can directly stimulate pancreatic beta-cells and indirectly enhance insulin resistance by suppressing insulin-mediated peripheral glucose uptake. Leptin activation seems to be similar in obese VPA-treated subjects to that seen in otherwise obese subjects. CONCLUSIONS: The mechanisms of hyperinsulinemia in VPA induced weight gain remain unclear, although it is likely that obesity is the cause of hyperinsulinemia and all related metabolic changes. However, this heterogeneous metabolic disorder requires further research. PMID- 19188737 TI - Mutations of the gene for the aryl hydrocarbon receptor-interacting protein in pituitary adenomas. AB - Heterozygous germline mutations in the gene encoding the aryl hydrocarbon receptor-interacting protein (AIP) were first described in two Finnish families with pituitary adenomas. The gene is involved in about 15% of familial isolated pituitary adenomas (FIPA), in about 50% of cases of familial acromegaly and in a small proportion of acromegalic patients with sporadic presentation. This review describes the genetic and clinical features of published patients with AIP, with either familial or sporadic pituitary tumors. A genotype-phenotype correlation is proposed: patients with AIP mutations resulting in a truncated protein are significantly younger than those bearing a mutation which preserves the structure of the C-terminal end of the protein (22.7 +/- 9.6 vs. 29.8 +/- 10.9 years). Pituitary tumors linked to AIP mutations are almost exclusively somatotropic (87.5%, n = 56/64) or lactotropic (9.4%, n = 6). Patients with AIP mutations are mostly men (70%, 44 M/19 F), suffer macroadenomas (97%) and are younger at diagnosis (24.4 +/- 10.5 years) than unselected patients with pituitary tumors. Thus, AIP is involved in the development of pituitary tumors, especially involving the somatomammotroph lineage. Genetic testing could be discussed for FIPAs and in young acromegalic patients with a sporadic presentation. Functional studies are needed to understand AIP-induced tumorigenesis. PMID- 19188738 TI - Short-term growth in children with congenital adrenal hyperplasia. AB - OBJECTIVE: To describe short-term growth patterns in children with congenital adrenal hyperplasia (CAH). METHODS: Height was measured daily in 5 children (1 boy) aged 3.9-9.7 years over 9-16 months. Kernel regression analysis was used to characterise short-term growth. The results were compared with data from 43 normal prepubertal children. RESULTS: Growth was characterised by growth spurts with intervening periods of no discernible growth (stasis). Height gain was positively correlated with the mean amplitude of growth spurts (r = 0.9, p < 0.05). Patients with CAH spent significantly less time in stasis than normal children (5 +/- 4.8 vs. 11.4 +/- 7.2% of study period; p < 0.05), the mean length of growth spurts was significantly longer (110.4 +/- 28.3 vs. 54.0 +/- 13.1 days; p < 0.05) and the mean amplitude significantly lower (0.022 +/- 0.008 vs. 0.037 +/- 0.001 cm/day; p < 0.01). CONCLUSIONS: Compared with normal controls, short term growth in children with CAH is characterised by long-duration low amplitude growth spurts with reduced periods of growth stasis. Better growth was correlated with the amplitude of growth spurts. The relatively smooth short-term growth in children with CAH suggests that if significant variations in growth rate are seen, they are more likely to be a consequence of under- or over-treatment rather than non-linear growth itself. PMID- 19188739 TI - Determinants of interleukin-6 and C-reactive protein vary in polycystic ovary syndrome, as do effects of short- and long-term metformin therapy. AB - OBJECTIVE, DESIGN AND METHODS: Circulating levels of interleukin-6 (IL-6) and C reactive protein (CRP) were determined in 59 women with polycystic ovary syndrome, of whom 37 were retested after receiving metformin for 6 weeks and 6 months, to ascertain the response of these inflammatory markers to weight loss and insulin sensitization. RESULTS: IL-6 levels correlated significantly with body mass index (BMI; r = 0.55), homoeostasis model insulin resistance index (HOMA-R; r = 0.47) and CRP (r = 0.59). On multiple regression analysis, adiposity rather than IL-6 accounted for most of the variability of CRP independently of insulin resistance, while IL-6 remained closely with adiposity. After 6 weeks of metformin therapy (n = 37) there was a significant reduction in BMI (p < 0.001), waist (p < 0.001) and CRP (p < 0.01), while at 6 months there was not only a significant reduction in BMI and waist but also in HOMA-R (p = 0.01) and IL-6 levels (p < 0.01) with no further reduction of CRP. CONCLUSIONS: In polycystic ovary syndrome, obesity rather than IL-6 or insulin resistance is the main determinant of CRP. While short-term metformin therapy facilitates weight loss with a concomitant reduction in CRP, long-term therapy results in a reduction of IL-6 and insulin resistance. Metformin-associated reduction of CRP levels prior to any significant changes in insulin resistance or IL-6 perhaps involves different mechanisms of action. PMID- 19188740 TI - Prevalence of thyroid dysfunction in obese children and adolescents before and after weight reduction and its relation to other metabolic parameters. AB - AIM: To establish the prevalence of elevated thyroid-stimulating hormone (TSH) levels in obese children and adolescents, and identify the relationship between changes in TSH levels and other metabolic and hormonal variables before and after weight reduction. METHODS: 207 obese participants aged 5-18 years were evaluated for anthropometric, biochemical, metabolic and hormonal variables before and after a weight reduction. RESULTS: At baseline, 46 participants (22.2%) had hyperthyrotropinemia (> or =4.0 mIU/l). Free T(4) levels were normal in all cases. Triglyceride levels were significantly higher in participants with hyperthyrotropinemia than in those with normal thyroid function (p = 0.011). Baseline TSH was significantly correlated with triglyceride levels (r = 0.261, p < 0.001), but not with age, anthropometric, or laboratory variables. Of the 142 participants who completed the intervention, 27 (19 %) had hyperthyrotropinemia. There was no significant relationship between changes in TSH level and changes in body mass index-standard deviation score. A significant correlation was found between the final TSH level and triglyceride level (r = 0.167, p = 0.045), and between the decrease in TSH level and the decrease in waist circumference (r = 0.291, p = 0.013). CONCLUSIONS: In obese children, hyperthyrotropinemia with normal free T(4) levels appears to be frequent. The correlation of hyperthyrotropinemia with waist circumference and higher triglyceride levels raises the question of the necessity to treat the elevated TSH levels. PMID- 19188741 TI - Glucocorticoid receptor gene polymorphism is less frequent in children born small for gestational age without catch-up growth. AB - BACKGROUND/AIMS: Glucocorticoids are important regulators of many processes involved in embryonal growth and development and fat and glucose metabolism. Glucocorticoids exert their effect through the glucocorticoid receptor (GR). The aim of this study was to investigate possible associations between 4 well-known GR gene haplotypes and size at birth. METHODS: We investigated associations between GR haplotypes and size at birth in a Dutch reference cohort. This reference cohort consisted of 222 young healthy Caucasian subjects. Associations between size at birth and glucocorticoid receptor gene haplotypes were tested. Furthermore, we investigated a group of 119 children born small for gestational age (SGA), without catch-up growth. Prevalence of the different GR haplotypes was compared between the SGA group and the reference cohort. RESULTS: No associations were found between any of the GR haplotypes and birth weight or birth length in the reference group. The prevalence of GR haplotype 2 (Bcl1) was significantly lower in the SGA group compared to controls. CONCLUSION: Genetic variance in the GR seems not to be associated with intrauterine growth in the general population. However, GR haplotype might play a role in growth of children born SGA, reflected by the decreased prevalence of GR haplotype 2 (Bcl1) in this group. PMID- 19188742 TI - Height gains in response to growth hormone treatment to final height are similar in patients with SHOX deficiency and Turner syndrome. AB - BACKGROUND: Patients with mutations or deletions of the Short Stature Homeobox containing(SHOX) gene have variable degrees of growth impairment, with or without mesomelic skeletal dysplasia. If untreated, short patients with SHOX deficiency remain short in adulthood. Growth hormone (GH) treatment improves short-term linear growth; however, there are no data on GH treatment effects on final height. PATIENTS: In a retrospective study, we assessed the relative effects of GH on final height gain in patients with SHOX deficiency (n = 14; 12 females) and Turner syndrome (TS) (n = 158). Patients were included if they fulfilled the following criteria: genetically-confirmed SHOX deficiency or TS, baseline height SDS <1.5, GH treatment started at Tanner stage < or =2, duration of GH treatment >2 years, and final height attained. RESULTS: Both groups of patients were short at baseline (height SDS [mean +/- SD]: SHOX deficiency, -3.3 +/- 0.9; TS, -2.9 +/ 0.8). Height SDS gain from baseline to final height was significant for each patient group (SHOX deficiency, 1.1 +/- 0.7; TS, 1.2 +/- 0.8; p < 0.001); however, it was not significantly different between groups (p = 0.708). CONCLUSIONS: Patients with SHOX deficiency receive similar final height benefit from GH treatment to those with TS. PMID- 19188743 TI - Analysis of craniofacial and extremity growth in patients with growth hormone deficiency during growth hormone therapy. AB - BACKGROUND/AIMS: There are many controversies regarding side effects on craniofacial and extremity growth due to growth hormone (GH) treatment. Our aim was to estimate GH action on craniofacial development and extremity growth in GH deficient patients. METHODS: Twenty patients with GH deficiency with a chronological age ranging from 4.6 to 24.3 years (bone age from 1.5 to 13 years) were divided in 2 groups: group 1 (n = 6), naive to GH treatment, and group 2 (n = 14), ongoing GH treatment for 2-11 years. GH doses (0.1-0.15 U/kg/day) were adjusted to maintain insulin-like growth factor 1 and insulin-like growth factor binding protein 3 levels within the normal range. Anthropometric measurements, cephalometric analyses and facial photographs to verify profile and harmony were performed annually for at least 3 years. RESULTS: Two patients with a disharmonious profile due to mandibular growth attained harmony, and none of them developed facial disharmony. Increased hand or foot size (>P97) was observed in 2 female patients and in 4 patients (1 female), respectively, both not correlated with GH treatment duration and increased levels of insulin-like growth factor 1. CONCLUSIONS: GH treatment with standard doses in GH-deficient patients can improve the facial profile in retrognathic patients and does not lead to facial disharmony although extremity growth, mainly involving the feet, can occur. PMID- 19188744 TI - Familial tumoral calcinosis caused by a novel FGF23 mutation: response to induction of tubular renal acidosis with acetazolamide and the non-calcium phosphate binder sevelamer. AB - Hyperphosphatemic familial tumoral calcinosis (HFTC) is an uncommon disease characterized by periarticular calcifications produced by the deposition of amorphous extraosseous calcifications of hydroxyapatite. It is associated with hyperphosphatemia due to increased tubular phosphate reabsorption, despite normal renal function and normal plasma PTH levels. The disease can be caused by inactivating mutations in either the fibroblast growth factor 23 (FGF23) gene, the UDP-N-acetyl-D-galactosamine:polypeptide N-acetylgalactosaminyltransferase 3 (GALNT3) gene or in human KLOTHO (KL) gene. Herein, we describe a Caucasian 3 year-old girl with tumoral calcinosis who presented with elevated serum phosphorus levels and a large calcified mass at her left elbow which led to ulceration of the skin. Treatment with the phosphate binder sevelamer and the carbonic anhydrase inhibitor acetazolamide successfully reduced the serum phosphate levels and led to a reduction of the calcified mass. This medical management has not been described previously. Her 7-month-old sister also had elevated serum phosphate levels, but did not have ectopic calcifications. Sequencing analysis revealed a novel homozygous FGF23 missense mutation (c.367G>T, p.Gly123Trp) in both siblings while the parents were carriers of the mutation. PMID- 19188745 TI - Research involving biological material from forensic autopsies: legal and ethical issues. AB - Recommendations and laws do not always contain specific and clear provisions on the use of cadaveric material in research, and even more rarely do they address explicitly the ethical issues related to research on material obtained during forensic autopsy. In this article we analyse existing legal frameworks in Europe by comparing the legal provisions in 2 European Countries which are member states of the Council of Europe, the UK and Switzerland. They were chosen because they have distinct legal frameworks that make comparisons interesting. In addition, the detailed laws of the UK and a specific law project and national ethical recommendations in Switzerland permit us to define more clearly the legal range of options for researchers using cadaveric material obtained during forensic investigations. The Human Tissue Act 2004 in England, Wales and Northern Ireland, its Scottish equivalent with the same title (2006) and the national ethical guidelines in Switzerland all require consent from the deceased person, an appropriate relative or a person with power of attorney for healthcare decisions before cadaveric biological material can be obtained and used for research. However, if the purpose of the autopsy is purely forensic, no such authorization will be sought to carry out the autopsy and related analyses, which might include genetic testing. In order to be allowed to carry out future research projects, families need to be approached for informed consent, unless the deceased person had left written directives including permission to use his or her tissues for research. PMID- 19188746 TI - Modulation of endothelium and endothelial progenitor cell function by low-density lipoproteins: implication for vascular repair, angiogenesis and vasculogenesis. AB - The endothelium regulates vascular homeostasis and is responsible for angiogenesis, a process mediated by the sprouting of endothelial cells from pre existing vessels. Several lines of evidence indicate that endothelial progenitor cells (EPCs) also play a role in adult neovascularization as well as in the maintenance of endothelial integrity and function. Hypercholesterolemia is associated with increased cardiovascular risk by inducing a cascade of events leading to endothelial dysfunction and injury. Growing evidence indicates that low-density lipoproteins (LDLs) impair endothelial reparative processes by inducing endothelial cell apoptosis but also by reducing the number and function of EPCs. The involvement of LDLs in mechanisms associated with vascular repair and neovascularization is also suggested by data from studies using lipid lowering drugs (statins). This review is focused on the central role of the cholesterol pathway in the biology of the endothelium and EPCs. PMID- 19188747 TI - Changes of the alternative splicing variants of human telomerase reverse transcriptase during gastric carcinogenesis. AB - OBJECTIVE: We attempted to reveal the changes of the human telomerase reverse transcriptase (hTERT) alternative splicing pattern in gastric carcinogenesis. METHODS: Three alternative splicing sites (alpha, beta, gamma) were selected and designed PCR primer. The expression of 8 hTERT alternative splicing variants (ASVs) in normal gastric mucosa, precancerous lesions and gastric cancer were detected by seminested RT-PCR. The expression of beta-site remaining ASV (beta(+) ASV) in specimens of precancerous lesions and specimens of gastric cancer was detected by SYBER Green real-time PCR. RESULTS: The positive rate of alpha(+)beta(+)gamma(+) ASV was significantly higher in gastric cancer than in precancerous lesions and normal mucosa (94.7 vs. 40.0% and 0%, p < 0.05). The positive rates of other ASVs were not different among the 3 groups (p > 0.05). The positive rates of beta(+) ASVs (including alpha(+)beta(+)gamma(+) ASV, alpha deletion ASV, gamma-deletion ASV, alphagamma-deletion ASV) were 11.1% in normal mucosa, 40.0% in precancerous lesions and 94.7% in gastric cancer (p < 0.05). SYBR Green real-time RT-PCR showed that the expression level of beta(+) ASV was 6.99 times higher in gastric cancer than in precancerous lesions. CONCLUSION: hTERT alternative splicing pattern is different during gastric carcinogenesis. beta(+) ASV was widely expressed in gastric carcinogenesis and may provide some information for diagnosis of gastric cancer or precancerous lesions. PMID- 19188749 TI - COX-2 expression does not correlate with microvessel density in breast cancer. AB - BACKGROUND: Cyclooxygenase-2 (COX-2), implicated in carcinogenesis and tumour progression in many cancers including breast cancer, is hypothesised to cause progression by promoting angiogenesis. The exact mechanism of such action is not known and the clinical evidence of such interaction is weak. We studied COX-2 expression and microvessel density (MVD) in malignant breast tissues. METHODS: COX-2 expression was analysed by immunohistochemistry in 89 breast cancer cases. MVD was assessed by CD31 immunohistochemistry using the Chalkey count method. COX 2 expression and MVD data were correlated with each other and with other prognostic factors. RESULTS: COX-2 expression, observed in 70 (79%) cases, correlated positively with tumour type (p = 0.037) and tumour grade (p = 0.045), but negatively with oestrogen receptor (p = 0.013). It did not correlate with tumour size, axillary lymph node status, progesterone receptor and HER-2 status. MVD varied from 2.09 to 40.38, correlated positively with tumour grade (p = 0.050) and tumour size (p = 0.044), but negatively with progesterone receptor (p = 0.040). MVD did not correlate with tumour type, axillary lymph node status, oestrogen receptor and HER-2. There was no correlation between COX-2 expression and MVD (p = 0.702). CONCLUSIONS: COX-2 expression does not correlate with angiogenesis in breast cancer. Angiogenesis in breast cancer may be dependent on multiple genes, rather than on COX-2 alone. PMID- 19188748 TI - Immunohistochemical analysis of neuroendocrine differentiation in prostate cancer. AB - OBJECTIVE: To clarify the significance of neuroendocrine differentiation in prostate cancer. METHODS: We immunohistochemically examined 96 samples of prostatic cancers obtained from radical prostatectomies using a specific neuroendocrine marker and various neuropeptides, as well as markers for cell proliferation, angiogenesis and androgen-receptor expression. RESULTS: We frequently found neuroendocrine cells in atrophic glands with or without chronic inflammation in nontumorous tissues. Neuroendocrine cells were detected in 36.5% of prostate cancer samples overall, but had no significant correlation to angiogenesis, cell proliferation or biochemical recurrence. However, patients with a high frequency of neuroendocrine cells (9.4%) tended to undergo preoperative hormonal therapy (p = 0.060), which led to their cancers being atrophic with inflammation. The neuroendocrine cells in these patients contained calcitonin-positive cells (p /=65 years old) with AMI admitted to Coronary Care Unit. METHODS: Elderly patients with AMI were retrospectively stratified for the presence and absence of PrA, and for quartiles of BMI and physical activity. In hospital outcomes (death, cardiogenic shock, and reinfarction and creatine kinase MB peak) were evaluated. RESULTS: In-hospital mortality of 1014 elderly patients with AMI was 19.2% in those with PrA and 22.7% in those without (P=0.18, NS). Mortality further decreased with increased physical activity and reduced BMI, a trend that was not observed in patients without PrA. When physical activity and BMI were considered together, lowest in-hospital mortality was observed in patients with highest physical activity and normal BMI (from 18.2 to 9.6%; P<0.01) with the greatest reduction observed in patients with PrA (from 18.3 to 5.1%; P=0.02). Multivariate analysis showed that PrA did not exert a protective effect in all patients irrespective of physical activity and BMI. A protective role was, however, observed in patients with highest physical activity or normal BMI and reached a maximum protective role in patients who showed both highest physical activity and normal BMI [odds ratio=0.08; 95% confidence interval=0.02 0.72; P<0.01]. CONCLUSION: The cardioprotective effect of PrA was preserved in elderly patients who showed the highest physical activity and a normal BMI. PMID- 19188808 TI - Blunted vascular responses but preserved endothelial vasodilation after submaximal exercise in chronic heart failure. AB - BACKGROUND: Patients with chronic heart failure (CHF) have abnormal vascular responses to acute exercise; however, regular aerobic training improves endothelial function in these patients. We hypothesized that, because of their attenuated vascular responses, CHF patients would present abnormal endothelium dependent vasodilatation after a single exercise session. METHODS: Thirteen CHF patients and 13 healthy controls participated in two experiments, on different days, using a cross-over design: (i) control (25-min seated at rest), and (ii) a submaximal exercise session (25-min cycling). Measurements of heart rate, blood pressure (BP), venous occlusion plethysmography forearm blood flow (FBF), and reactive hyperemia were made before and after (immediately, 10, 30, 60 min, and 24 h) each experimental condition. RESULTS: CHF patients had no changes in the mean BP throughout the protocols, whereas mean BP was reduced up to 60 min after exercise in controls. In CHF patients, FBF was increased and forearm vascular resistance was reduced up to 10 min after exercise, whereas these changes were sustained up to 30 min after exercise in controls. Reactive hyperemia was significantly increased up to 30 min after exercise in both groups. CONCLUSION: Patients with CHF have increased postexercise FBF and decreased forearm vascular resistance; however, these responses last longer in healthy individuals. Despite the attenuated postexercise vascular responses, patients with CHF respond to a single-cycle exercise session with improved forearm endothelium-dependent vasodilation. PMID- 19188810 TI - Prevalence, awareness, treatment and control of high blood pressure in a Swiss city general population: the CoLaus study. AB - BACKGROUND: This study is aimed to assess the prevalence of awareness, treatment and control of high blood pressure (HBP) and associated factors in a Swiss city. DESIGN: Population-based cross-sectional study of 6182 participants (52.5% women) aged 35-75 years living in Lausanne, Switzerland. METHODS: HBP was defined as blood pressure >/=140/90 mmHg or current antihypertensive medication. RESULTS: The overall prevalence of HBP was 36% (95% confidence interval: 35-38%). Among participants with HBP, 63% were aware of it. Among participants aware of HBP, 78% were treated, and among those treated, 48% were controlled (BP <140/90 mmHg). In multivariate analysis, HBP prevalence was associated with older age, male sex, low educational level, high alcohol intake, awareness of diabetes or dyslipidaemia, obesity and parental history of myocardial infarction. HBP awareness was associated with older age, female sex, awareness of diabetes or dyslipidaemia, obesity and parental history of myocardial infarction. HBP control was associated with younger age, higher educational level and no alcohol intake. Alone or in combination, sartans were the most often prescribed antihypertensive medication category (41%), followed by diuretics, beta-blockers, angiotensin converting enzyme inhibitors and calcium channel blockers. Only 31% of participants treated for HBP were taking >/=2 antihypertensive medications. CONCLUSION: Although more than half of all participants with HBP were aware and more than three-quarters of them received a pharmacological treatment, less than half of those treated were adequately controlled. PMID- 19188809 TI - Outcome of patients with acute myocardial infarction admitted in hospitals with or without catheterization laboratory: results from the HELIOS registry. AB - AIMS: To compare the treatment and outcomes of myocardial infarction patients in hospitals with and without catheterization laboratory. METHODS AND RESULTS: The Hellenic Infarction Observation Study was a countrywide registry of acute myocardial infarction, conducted during 2005-2006. The registry enrolled 1840 patients with myocardial infarction from 31 hospitals with a proportional representation of all types of hospitals and of all geographical areas. Out of these patients, 645 (35%) were admitted in 11 hospitals with and 1195 (65%) in 20 hospitals without catheterization laboratory. Patients admitted in hospitals with catheterization laboratory in comparison with patients admitted in hospitals without were younger (66+/-14 vs. 68+/-13, P<0.004) with less diabetes (27 vs. 33%, P<0.001), but without other baseline differences (female 27 vs. 25%, prior myocardial infarction 20 vs. 17%, Killip class>1 22 vs. 23%). Reperfusion rates for ST-segment elevation myocardial infarction were 67% (43% lytic, 24% primary percutaneous coronary interventions) versus 56% (55% lytic, 1% percutaneous coronary interventions; P<0.01). In-hospital outcomes in hospitals with versus in hospitals without laboratory were: mortality 6.5 versus 8.3% (NS), stroke 2.2 versus 1.1% (NS), major bleeding 1.1 versus 0.6% (NS), and heart failure 11 versus 16% (P<0.01). In multivariate regression analysis, being admitted in a hospital without catheterization laboratory was not an independent predictor of increased in-hospital mortality (odds ratio=1.18, 95% confidence interval: 0.72 1.93, P=0.505). CONCLUSION: Although the majority of acute myocardial infarction patients was admitted in hospitals without catheterization laboratory, these patients do not have a survival disadvantage, provided they are treated with lytic therapy, medical secondary prevention drugs, and eventual revascularization according to current guidelines. PMID- 19188811 TI - The association of serum lipids with renal function: the Korea Medical Institute Study. AB - BACKGROUND: Both serum lipids and renal dysfunction are well-known risk factors for cardiovascular diseases. Little data are, however, available on the relation of serum lipids and glomerular filteration rates (GFRs) with low levels of serum cholesterol in Asian populations. METHODS: We performed a large cross-sectional study of 93 228 Korean men and women enrolled in the Korea Medical Institute Study. Serum creatinine was used to estimate the GFR through the simplified modification of diet in renal disease equation. RESULTS: In multivariate logistic regression analysis, those in the highest quartiles of all the lipid biomarkers showed significant associations with the risk of having a GFR of less than 60 ml/min per 1.73 m, compared with those in the lowest quartiles except high density lipoprotein (HDL) in women. In men, odds ratios (ORs) by triglycerides (TGs) [OR: 1.99; 95% confidence interval (CI): 1.65-2.40], and TG/HDL (OR: 1.98; 95% CI: 1.64-2.38) were higher than others. In women, low-density lipoprotein (OR: 1.91; 95% CI: 1.42-2.56) and non-HDL (OR: 1.84; 95% CI: 1.35-2.50) showed the highest ORs. These associations were stronger among male participants with diabetes; total cholesterol (OR: 2.03; 95% CI: 1.04-3.96), TG (OR: 18.08; 95% CI: 2.46-133.13) and TG/HDL (OR: 17.97; 95% CI: 2.44-132.30), respectively. CONCLUSION: We conclude that potentially modifiable lipid biomarkers are elevated in the setting of less than 90 ml/min per 1.73 m of GFR and there will be the link between renal insufficiency and the increased risk for cardiovascular events in the Korean population. PMID- 19188812 TI - SRY gene increases the risk of developing gonadoblastoma and/or nontumoral gonadal lesions in Turner syndrome. AB - The presence of Y-chromosome material in patients with dysgenetic gonads increases the risk of gonadal tumors and/or nontumoral androgen-producing lesions. The patients' prognosis can vary, depending on their karyotype. The objective of this study was to investigate the presence of Y-chromosome mosaicism in Turner syndrome patients and its association with the development of gonadal tumors and/or nontumoral androgen-producing lesions. Eighty-seven Turner syndrome patients were studied. Genomic DNA was extracted from peripheral blood and genes SRY and TSPY and DYZ3 repeat of the Y chromosome were amplified by polymerase chain reaction. To the Y-positive patients, prophylactic gonadectomy was offered. The data disclosed hidden Y-chromosome mosaicism in 16 (18.5%) of the patients. SRY sequence was detected in all of the 16 patients, and 4 (4.6%) of them presented DYZ3 repeat region and TSPY gene. Eleven of the patients with Y positive sequences agreed to undergo the prophylactic surgery. In 2 cases, bilateral gonadoblastoma was found and, in another case, the histopathologic study of the gonads revealed hilus cell hyperplasia. In a further case, there were hilus cell hyperplasia and a stromal luteoma. In conclusion, a systematic search for hidden Y-chromosome mosaicism, especially SRY, in Turner syndrome patients is justified by the possibility of preventing gonadal lesions. PMID- 19188813 TI - Ovarian and paraovarian squamous-lined cysts (epidermoid cysts): a clinicopathologic study of 18 cases with comparison to mature cystic teratomas. AB - The origin of ovarian epidermoid cyst is unknown. Sixteen ovarian and 2 paraovarian squamous-lined cysts unassociated with teratomatous elements were studied. The ovarian cysts represented 1.5% of consecutive ovarian surface epithelial tumors examined and were one-nineteenth as common as mature cystic teratoma. The mean patient age was 57 years. All tumors were unilateral and the majority were incidental findings. The mean tumor size was 1.75 cm and the median, 3.0 cm. In comparison to 120 consecutive patients with mature cystic teratoma who had a mean age of 41 years and a mean tumor size of 6 cm, the ovarian epidermoid cysts were significantly smaller and occurred at a significantly older age (P<0.01). All cysts displayed mature squamous epithelium with a granular layer, with hyperkeratosis in 14 and parakeratosis in 4. In 2 patients, there were contralateral mature cystic teratomas, and in 2 others the cysts contained rare hairs. Two displayed foci of Brenner tumor and 1 appeared to arise in endometriosis. In summary, 7 of 16 ovarian epidermoid cysts displayed features suggesting they reflected insufficiently sampled teratomas, Brenner tumors with squamous metaplasia, or a metaplastic change in endometriosis. Epidermoid cyst of the ovary as defined by histology is a heterogeneous group; pure epidermoid cyst, if it exists at all, probably represents less than 1% of ovarian surface epithelial tumors. PMID- 19188814 TI - Fascin expression in serous tumors of ovary correlates with aggressiveness of malignancy. AB - Ovarian serous tumors make up about one-fourth of all ovarian tumors. There is a spectrum of proliferation and cellular atypia in these tumors with benign serous cystadenoma, borderline tumors, and low grade or type I serous carcinoma at the lower end and type II or high-grade serous papillary cystadenocarcinoma at the higher end. Fascin is a globular actin cross-linking protein involved in cell motility that has been shown to be upregulated in many human neoplasms and associated with the aggressiveness of malignancy. The aim of this study was to investigate fascin expression in serous tumors of ovary and to evaluate its relationship with the aggressiveness of tumor. Sections from a total of 66 serous tumors of ovary were collected including 26 serous carcinomas, 20 borderline serous tumors, and 20 benign serous cystadenomas. Ten benign ovaries with inclusion cysts were used as controls. Sections were immunostained with fascin. Fascin expression was significantly increased in borderline (13/20, 65%) and malignant serous tumors (22/26, 84%) compared with benign serous cystadenoma (0/20) (P<0.001). There was increased quantitative expression of fascin in carcinoma compared with borderline tumors (diffuse versus patchy). Fascin expression also correlated well with the tumor grade in serous carcinoma cases with 8/12 (66%) of grade I/II tumors staining positive compared with all 14 (100%) of grade III tumors showing fascin expression (P<0.05). Our findings suggest that upregulation of fascin plays a role in increasing aggressiveness of serous ovarian tumors and could potentially be a molecular therapeutic target and a prognostic marker. PMID- 19188815 TI - Increased p16 expression in high-grade serous and undifferentiated carcinoma compared with other morphologic types of ovarian carcinoma. AB - There are several morphologic types of ovarian carcinoma. It has been shown that p16 is overexpressed in high-grade serous carcinoma but there has been little detailed comparison of p16 expression in the common types of ovarian carcinoma. The aim of this study was to compare p16 expression in ovarian carcinomas of serous, endometrioid, clear cell, and mucinous type with a view to ascertaining whether high expression in a primary ovarian carcinoma is specific for a serous neoplasm. We included problematic cases, which are difficult to type, such as poorly differentiated and undifferentiated carcinomas and serous carcinomas with clear cells. In these problematic groups, we compared p16 expression with that of WT1, which is known to be relatively specific for a serous phenotype. Cases of ovarian high-grade serous carcinoma (n=38), endometrioid carcinoma (n=15), clear cell carcinoma (n=12), and mucinous carcinoma (n=10) were stained with p16. Cases were scored both with respect to distribution of immunoreactivity (0-5) and intensity (0-3). An immunohistochemical composite score was also calculated (0 15) by multiplying the distribution and intensity scores. Serous carcinomas typically exhibited high p16 expression; there was statistically significant higher p16 expression in serous carcinomas compared with the other morphologic types. There was high p16 and WT1 expression in most undifferentiated carcinomas and in serous carcinomas with clear cells, suggesting that these represent variants of serous carcinoma. We have demonstrated that p16 is highly expressed in high-grade serous and undifferentiated carcinomas compared with other morphologic types of ovarian carcinoma. This may be useful, in conjunction with WT1, in the classification of problematic neoplasms. p16 may be involved in the pathogenesis of high-grade ovarian serous carcinomas, possibly through inactivation of retinoblastoma protein. PMID- 19188816 TI - P63 expression in gestational trophoblastic disease: correlation with proliferation and apoptotic dynamics. AB - p63 regulates cell growth and differentiation and contributes to tumorigenesis through its complex isoforms. Gestational trophoblastic disease encompasses a heterogeneous family of lesions with different malignant potential that arise from various trophoblast subpopulations. This study investigated the expression of p63 isoforms in various trophoblastic diseases and correlated with clinical progress, proliferation, and apoptotic activities. 4A4 and anti-p40 antibodies were applied to assess expressions of total and DeltaNp63 isoforms in 20 placentas, 62 hydatidiform moles, 9 choriocarcinomas, 5 placenta site trophoblastic tumors, and 2 epithelioid trophoblastic tumors immunohistochemically. The immunoreactivity of p63 was localized to the nuclei of cytotrophoblast, villous, and chorionic-type intermediate trophoblasts with significant correlation between 2 p63 indices (P<0.001). p63 indices were significantly lower in placentas of advanced gestational age (P<0.001). Hydatidiform moles demonstrated significantly higher p63 indices than normal placentas (P<0.001). Epithelioid trophoblastic tumors displayed the highest p63 indices (45%-80%) whereas immunoreactivity was only focal in choriocarcinoma (0% 5.62%) and was essentially absent in placenta site trophoblastic tumors. There was no significant correlation between p63 indices and subsequent development of trophoblastic neoplasia in hydatidiform moles (P>0.05). Both p63 indices positively correlated with the proliferative index (Ki67) (P<0.05), apoptotic index (M30) (P<0.005), p53 (P<0.005), and p21(WAF1/CIP1) expression (P<0.005). Our results indicate that DeltaNp63, the dominant isoforms expressed in trophoblasts, display heterogeneous expression patterns in relation to trophoblast subtypes. We also demonstrate for the first time the possible role of p63 in the pathogenesis of gestational trophoblastic disease (GTD) through its interaction with p53-dependent proliferation and apoptotic activities. PMID- 19188817 TI - Myometrial myxoidosis: a report of 2 cases of a distinctive type of secondary myometrial hypertrophy in patients with lupus erythematosus. AB - Myxoid mesenchymal lesions of the uterus are generally restricted to tumors, but non-neoplastic myxoid mesenchymal lesions of the uterus have not received much attention in the literature. We analyzed the clinicopathologic features of 2 patients with lupus erythematosus (ages 43 and 52 yr, respectively) in whom myometrial myxoidosis produced a markedly enlarged uterus with myometrial thickening ("secondary myometrial hypertrophy"). Both patients underwent a hysterectomy for presumed leiomyomas, and intraoperatively an enlarged uterus was noted. On gross examination, the uteri measured 13.5 x 13.5 x 11.5 cm and 14.5 x 11.5 x 9.5 cm, respectively. The significantly thickened myometrium was due to marked expansion of the interstitial compartment of the myometrium, in which non neoplastic smooth muscle fascicles were widely separated by abundant extracellular mucin producing a striking myxoid appearance ("myxoidosis"). These histologic findings are akin to the pattern of dermal mucin deposition seen in lupus erythematosus. The lesion in each case diffusely involved the entire myometrium. Histochemical stains were performed and showed the following results: mucicarmine-diffusely but weakly positive; periodic acid-schiff (PAS)-negative; colloidal iron-diffuse positive; alcian blue, pH 2.5 (without hyaluronidase digestion)-diffuse positive, and alcian blue, pH 2.5 (with hyaluronidase digestion)-negative. These histochemical findings are consistent with hyaluronic acid. Follow-up in 1 case was not available. In the other case, the patient presented to clinical attention 5 weeks after surgery because of ascites, which after an extensive clinical evaluation was interpreted as being of unknown etiology. To the best of our knowledge, this rare and unusual non-neoplastic myometrial lesion has not been previously described. Pathologists should be aware of its existence because of the distinctive appearance and as it may prompt consideration of various myxoid neoplasms of the uterus in the differential diagnosis. Patients with myometrial myxoidosis should be evaluated for lupus erythematosus. PMID- 19188818 TI - Expression of vascular endothelial growth factor (VEGF), hypoxia inducible factor 1alpha (HIF-1alpha), and microvessel density in endometrial tissue in women with adenomyosis. AB - Adenomyosis is a disease with a mysterious pathogenesis, defined by an abnormal displacement of the eutopic endometrium deeply and haphazardly inside the myometrium. Angiogenesis has been indicated to play an important role and our aim was to investigate whether vascular endothelial growth factor (VEGF) and hypoxia inducible factor-1alpha (HIF-1alpha) expression and microvessel density (MVD) were different in women with and without adenomyosis. Immunohistochemistry was performed in endometrial tissues in 23 patients who underwent radical hysterectomy for adenomyosis (14) and for ovarian cysts and fibroids (9) at an Academic Hospital. Compared to women without the disease, VEGF expression was increased in endometrium with a normal location in patients with adenomyosis, although not associated to a significant increase of HIF-1alpha and MVD. Moreover, the endometrium with an abnormal location in patients with adenomyosis showed an increased VEGF and HIF-1alpha expression, particularly in the epithelial cells, associated to an increase of MVD, compared with the endometrium in a normal location in the same group of patients. Our present findings suggest that VEGF-mediated angiogenesis might be associated with the development of adenomyosis. In the ectopic foci the abnormal location might contribute to increased HIF-1a expression, stimulation of VEGF production, and increased vessel formation. In endometrium with a normal location, instead, where VEGF increased expression seems not to be correlated with HIF-1alpha increased expression nor with an increased MVD, other mechanisms might be reasonably postulated. Additional studies are required to explore new targeted and more effective treatment modalities. PMID- 19188819 TI - Expression of cyclooxygenase-2 and matrix metalloproteinase-2 in adenomyosis and endometrial polyps and its correlation with angiogenesis. AB - This study investigates the expression of cyclooxgenase (COX)-2 and matrix metalloproteinase (MMP)-2 in patients with adenomyosis or endometrial polyps and their possible relation to microvascular density in these lesions. The subjects were 25 patients with adenomyosis, 30 patients with endometrial polyps, and 20 female controls. The expression of COX-2, MMP-2, and CD34 was studied immunohistochemically. Microvesseldensity (MVD) was calculated by the counting of CD34-positive vascular endothelial cells. The quantity and intensity of COX-2 expression in endometrium did not vary during the menstrual cycle in the control group and in patients with endometrial polyps. In patients with adenomyosis, it was higher in the secretory phase. MMP-2 expression in stromal cells in adenomyotic foci and endometrial polyps were higher than in normal endometrium. In the proliferative phase, MVD in adenomyosis foci was higher than in normal endometrium and endometrial polyps. In the secretory phase, MVD in adenomyotic foci and endometrial polyps was higher than in normal endometrium. Overexpression of stromal MMP-2 may play a role in the development of adenomyosis and endometrial polyps. Aberrant COX-2 expression in eutopic endometrium during the luteal phase may be associated with the pathogenesis of adenomyosis; however, expression of COX-2 does not seem to play a role in the development of endometrial polyps. MVD was high in both lesions, but there was no significant correlation between MVD and the expression of MMP-2 or COX-2. Mechanisms other than COX-2 and MMP-2 may contribute to the promotion of angiogenesis in these lesions. PMID- 19188820 TI - The significance of neuroendocrine expression in undifferentiated carcinoma of the endometrium. AB - Carcinomas with neuroendocrine (NE) differentiation have been associated with poor outcome in different organs. The purpose of this study was to evaluate the presence and significance of NE expression in a series of 46 undifferentiated endometrial carcinomas diagnosed between 1988 and 2005. NE expression was studied by immunohistochemistry including synaptophysin, chromogranin, and/or CD56. The patients' age ranged from 30 to 84 years (mean 55). Staging information was available for 45 cases and according to the International Federation of Gynecology and Obstetrics system they were distributed as follows: stage I (9 cases), stage II (2 cases), stage III (9 cases), and stage IV (25 cases). NE expression was present in 19/46 (41%) cases; however, it was diffuse in only 9% of the tumors. The median survival for patients without NE expression was 7 months (95% confidence interval 4-10 mo) and for patients with NE expression was 12 months (95% confidence interval 6-27 mo). The survival curves do not differ significantly (P=0.49). NE expression is common in undifferentiated carcinoma of the endometrium, as it was found in 41% of our cases. In most cases, NE expression is only focally present (< or =10% of the cells). There is no difference in overall survival in patients with or without NE expression. PMID- 19188821 TI - Endocervical adenocarcinoma in situ, serous type. PMID- 19188822 TI - Metastatic HPV-related cervical adenocarcinomas presenting with thromboembolic events (Trousseau Syndrome): clinicopathologic characteristics of 2 cases. AB - Two cases of systemic thromboembolism (Trousseau syndrome) associated with metastatic human papillomavirus (HPV)-related endocervical adenocarcinomas are reported. The first patient, age 36, presented with bilateral lower extremity deep vein thromboses, pulmonary embolism, and supraclavicular and cervical lymphadenopathy. Lymph node biopsy revealed metastatic mucinous adenocarcinoma with focal signet ring cell differentiation. Imaging studies demonstrated metastatic disease without a defined primary site. Acute renal and respiratory failure developed and the patient expired shortly after initiation of chemotherapy, 7 weeks after presentation. Autopsy examination revealed widespread metastatic adenocarcinoma with a 2 cm cervical adenocarcinoma. The second patient, age 43, presented with left internal jugular vein thrombosis, acute thrombophlebitis, and bilateral axillary lymphadenopathy. She developed progressive venous thrombosis despite anticoagulation. Imaging studies demonstrated widespread lymphadenopathy and an adnexal mass. Diagnostic laparoscopy with biopsies and left oophorectomy revealed metastatic mucinous adenocarcinoma with signet ring cell differentiation involving peritoneum, ovary, cervix, and bladder without a defined primary site. Progressive thromboembolic disease with acute renal failure and multiple cerebral infarcts developed and the patient expired shortly after initiation of chemotherapy, 2 months after presentation. No autopsy was performed. HPV DNA was detected by in situ hybridization in the lymph node metastasis in the first case and in the cervical and ovarian tumor specimens in the second case. High-risk HPV-related endocervical adenocarcinomas occasionally exhibit signet ring cell differentiation and can present with Trousseau syndrome. These features more commonly suggest metastatic adenocarcinoma of upper gastrointestinal tract origin but the presence of HPV DNA within the tumors establishes them as cervical in origin. PMID- 19188823 TI - Upregulation of MUC4 in cervical squamous cell carcinoma: pathologic significance. AB - MUC4 is a transmembrane glycoprotein more highly expressed in cervical dysplasia than benign cervical epithelium. We sought to determine whether MUC4 expression differs between benign and malignant cervical tissue. Fifty-eight patients with benign, dysplastic, or malignant cervical pathology were identified retrospectively, and representative sections were stained with a mouse monoclonal anti-MUC4 antibody. Semiquantitative analysis was performed on benign, dysplastic, and malignant regions by scoring staining intensity (0: negative, 1: weak, 2: moderate, and 3: strong) and distribution (focal <10%, multifocal=10% 60%, diffuse > or =60%). In samples with benign glycogenated squamous epithelium, only the parabasal cells had MUC4 staining, and 48.5% had an intensity of 2 or 3. All samples with immature squamous metaplasia were positive through the entire epithelial thickness. Cervical intraepithelial neoplasia (CIN) 1 samples had variable staining with an intensity similar to glycogenated squamous epithelium but distribution similar to squamous metaplasia. All CIN 3 (n=21) and invasive squamous cell carcinomas (n=17) had increased MUC4 staining intensity (P<0.001 and P<0.001) and increased diffuse staining (P<0.001 and P<0.001) compared with the limited staining in glycogenated squamous epithelium. In contrast, no differences in staining were observed between benign endocervical glands, adenocarcinoma in situ, and invasive adenocarcinoma. These expression patterns suggest that MUC4 is a lineage marker in benign cervical tissue that may have aberrant expression in squamous dysplasia and carcinoma. Further studies may elucidate the role of MUC4 in the development of squamous cell cervical cancer. PMID- 19188824 TI - HPV determination in the control after LEEP due to CIN II-III: prospective study and predictive model. AB - To evaluate the usefulness of the endocervical cytology immediately after loop electrosurgical excision procedure (LEEP) and viral human papilloma virus (HPV) determination as predictive factors of persistence/recurrence (P/R) of squamous intraepithelial lesion (SIL) in the remaining cervix of patients treated with LEEP due to cervical intraepithelial neoplasia (CIN) II or III. We retrospectively selected 105 samples from 100 patients with histologic diagnosis of CIN II-III with cytology immediately after LEEP with a first control post-LEEP between 2 and 6 months after treatment that included HPV determination, a minimal follow-up period of 12 months and maximum of 24 months, and at least 2 colpocytologic controls post-LEEP. In 71 of the 105 patients (67.6%), the determination was negative, finding the presence of HPV in 34 patients (32.4 %). Almost two-thirds of the patients had negative endocervical cytology and absence of HPV. During the follow-up, 20 P/R were detected, 16 (15.2%) of them being high grade squamous intraepithelial lesions (7 CIN II and 9 CIN III). The combination of endocervical cytology and HPV determination in the remaining cervix seems to be a good strategy to predict the risk of SIL after conization. The probability of P/R can be estimated based on the results of these 2 variables. The negativity of HPV determination together with a negative endocervical cytology guarantees enough security and a favorable evolution. PMID- 19188825 TI - A pilot evaluation of a novel immunohistochemical assay for topoisomerase II alpha and minichromosome maintenance protein 2 expression (ProEx C) in cervical adenocarcinoma in situ, adenocarcinoma, and benign glandular mimics. AB - The histopathologic distinction of cervical adenocarcinoma in situ (AIS) and invasive adenocarcinoma (AC) from some benign endocervical lesions can be challenging. The ProEx C antibody reagent targets nuclear proteins (minichromosome maintenance protein 2, MCM2 and topoisomerase II-alpha, TOP2A), which are over expressed during the aberrant S-phase induction of HPV infected and neoplastic cells. In this immunohistochemical study the utility of the ProEx C reagent in distinguishing AIS and AC from a variety of non-neoplastic glandular lesions was examined. ProEx C immunohistochemical staining was performed on sections from formalin-fixed, paraffin-embedded tissue of 65 cervical tissues including 48 non-neoplastic cervices (normal [n=10], microglandular hyperplasia [n=10], tubal metaplasia [n=11], cervical endometriosis [n=7], reactive endocervix [n=10]) and 17 cervices with glandular malignancy (AIS [n=12] and AC [n=5]). Both intensity and prevalence of immunoreactivity was scored. The median and distribution of scores for both prevalence and intensity was compared for AIS versus each of the 5 benign cervical lesions using a Mann-Whitney U test. The median and distribution of prevalence of immunohistochemical staining for AIS was different from all benign mimics, but the intensity of staining for AIS did overlap with some mimics as it was not significantly different from endometriosis, microglandular hyperplasia, and reactive endocervix. ProEx C reagent has potential as an adjunctive testing tool in the histopathologic diagnosis of both AIS and AC, particularly in difficult cases with small biopsies or foci of disease. PMID- 19188826 TI - P16 immunostaining patterns in microglandular hyperplasia of the cervix and their significance. AB - P16 immunostaining is an important adjunct in the differential diagnosis of difficult squamous and glandular intraepithelial lesions of the cervix. However, unexpected staining of epithelium other than the target lesion can pose a problem in the interpretation. This study examined a common entity in the cervix, microglandular hyperplasia (MGH), that is associated with proliferations of both columnar and squamous epithelial cells-and ascertained the frequency of p16 staining, its pattern, and relationship to human papillomavirus. Fifty-seven cases of MGH were analyzed; 25 scored strongly immunopositive (44%). In 18, staining of the superficial columnar epithelium was patchy, involving 10% to 20% of cells on the surface; in 4 cases, 30% to 40% of cells; and in another 3, over 50% of the cells in a given area were strongly positive. Staining involved both nucleus and cytoplasm of columnar cells. P16 positivity did not colocalize with either cyclin E or MIB-1. Adjacent non-MGH-related columnar epithelium scored negative for p16. Of 25 p16-positive columnar epithelia analyzed, all were human papillomavirus -negative. In conclusion, benign columnar epithelium in the setting of MGH can be expected to stain strongly for p16. Practitioners should be aware of this when evaluating diagnostically difficult squamous or glandular epithelial changes occurring in the setting of MGH or when interpreting cytologic preparations stained with p16. PMID- 19188827 TI - History of gynecological pathology: XXII. Dr. Johann Veit, M.D. PMID- 19188828 TI - Effects of combined treatment with angiotensin II type 1 receptor blocker and statin on stent restenosis. AB - Angiotensin II type I receptor blocker (ARB) or statin has been reported to be effective in preventing stent restenosis, but little is known about the combined effect on stent restenosis. The objective of this study was to determine the effect of combination therapy with ARB and statin on restenosis rate after coronary stenting and on proliferation and migration of and reactive oxygen species (ROS) production by human coronary artery smooth muscle cells (SMCs) in vitro. Clinical data were collected from 330 consecutive patients who underwent coronary stenting for de novo lesions. Six months after stenting, quantitative coronary angiography was performed. Combined therapy with the ARB and statin significantly inhibited stent restenosis (P < 0.05) compared with the effect of the ARB or statin alone. In an in vitro study, platelet-derived growth factor (PDGF)-induced proliferation was significantly inhibited by combined treatment with CV11974, an ARB, and simvastatin (P < 0.01), but the inhibitory effect was not significantly greater than that of simvastatin alone. Migration of human coronary artery SMCs was significantly inhibited by the ARB + statin compared with the effect of the ARB or statin alone (P < 0.01). PDGF-induced production of ROS was also inhibited significantly by the ARB + statin compared with the effect of the ARB or statin alone (P < 0.01). These results indicate that inhibitory effects of combined therapy on PDGF-induced migration of and ROS production by SMCs play an important role in reduction of restenosis rate. Combined treatment with ARB and statin after stenting is useful for preventing stent restenosis. PMID- 19188829 TI - ETA receptor blockade with atrasentan prevents hypertension with the multitargeted tyrosine kinase inhibitor ABT-869 in telemetry-instrumented rats. AB - ABT-869 is a novel multitargeted inhibitor of vascular endothelial growth factor and platelet-derived growth factor receptor tyrosine kinases (RTKs) with potent antiangiogenic properties that slow tumor progression. Vascular endothelial growth factor receptor blockade has been shown to produce hypertension. Atrasentan is a potent and selective endothelin (ETA) receptor antagonist that lowers blood pressure and affects tumor growth. To assess the utility of ETA receptor blockade in controlling hypertension with RTK inhibition, we evaluated the ability of atrasentan to block hypertension with ABT-869 in conscious, telemetry-instrumented rats. Changes in mean arterial pressure (MAP) and heart rate (HR) were evaluated using mean values and the area under the curve (AUC). Atrasentan (0.5, 1.5, and 5.0 mg kg(-1) d(-1) for 5 days) elicited dose-dependent decreases in MAP-AUC (-16.7 +/- 1.3, -20.94 +/- 3.68, and -30.12 +/- 3.57 mm Hg x day, respectively) compared with vehicle. ABT-869 (1, 3, 10, 30 mg kg(-1) d(-1) for 5 days) increased MAP compared with vehicle (MAP-AUC values of -5.52 +/- 3.75, 12.7 +/- 8.4, 37.5 +/- 4.4, and 63.8 +/- 3.3 mm Hg x day, respectively). Pretreatment with atrasentan (5 mg/kg for 5 days) prevented and abolished the hypertensive effects of ABT-869. Thus, ETA receptor blockade effectively alleviated hypertension with RTK inhibition and may serve a dual therapeutic role by preventing hypertension and slowing tumor progression. PMID- 19188830 TI - Involvement of NO and KATP channel in adenosine A2B receptors induced cardiovascular regulation in the posterior hypothalamus of rats. AB - Previous reports have suggested that the posterior hypothalamic adenosine A2 receptors may play a role in central cardiovascular regulation. In this study, we examined the influence of posterior hypothalamic adenosine A2B receptors on the regulation of blood pressure and heart rate. Drugs were injected into the posterior hypothalamus of anesthetized, artificially ventilated, male Sprague Dawley rats. Four nanomoles of 5'-N-ethylcarboxamidoadenosine (NECA), an adenosine A 2A receptor agonist, decreased arterial blood pressure and heart rate, whereas 5 nmol of alloxazine, an adenosine A2B receptor antagonist, blocked the depressor and bradycardiac effects of 4 nmol NECA. We examined the role of nitric oxide (NO) and K+ channels on cardiovascular regulation by adenosine A2B receptors in the posterior hypothalamus. Pretreatment with 40 nmol of NG-nitro-L arginine methyl ester, a NO synthase inhibitor, significantly attenuated the effects of NECA, and 10 nmol of sodium nitroprusside, a NO releaser, strengthened the action of drug. In addition, posterior hypothalamic administration of 20 nmol of glipizide, an K ATP blocker, blocked the cardiovascular depression elicited by NECA. These results suggest that NO mediates cardiovascular regulation by activation of A2B receptors in the posterior hypothalamus. Additionally, ATP sensitive K+ channels modulate the action of adenosine A2B receptors. PMID- 19188831 TI - Mechanisms of cardiac muscle insensitivity to a novel acetylcholinesterase inhibitor C-547. AB - We compared the effects of the novel acetylcholinesterase (AChE) inhibitor C-547 on action potential configuration and sinus rhythm in the isolated right atrium preparation of rat with those of armin and neostigmine. Both armin (10(-7), 10( 6), and 10(-5) M) and neostigmine (10(-7), 10(-6), and 5 x 10(-6) M) produced a marked decrease in action potential duration and slowing of sinus rate. These effects were abolished by atropine and are attributable to the accumulation of acetylcholine in the myocardium. The novel selective AChE inhibitor C-547 (10(-9) to 10(-7) M), an alkylammonium derivative of 6-methyluracil, had no such effects. The inhibition constant of C-547 on cardiac AChE is 40-fold higher than that on extensor digitorum longus muscle AChE. These results suggest that C-547 might be employed to treat diseases such as myasthenia gravis or Alzheimer disease, without having unwanted effects on the heart. PMID- 19188832 TI - Intermittent inotropic infusions combined with prophylactic oral amiodarone for patients with decompensated end-stage heart failure. AB - BACKGROUND: Concern has been raised regarding the mortality and ethics related to the treatment of patients with end-stage chronic heart failure with chronic intermittent intravenous inotropic agents. We examined whether intermittent inotropic agents combined with oral amiodarone to prevent the proarrhythmic effect of inotropic agents results in better outcomes. METHODS: The study included 162 patients with decompensated end-stage chronic heart failure, who could be weaned from an initial 72-hour infusion of intravenous inotropes. Group 1 included 140 patients, who entered a 6-month program of weekly intermittent intravenous inotropic agents plus oral amiodarone, 200 mg twice a day. Group 2 included 22 patients, who were treated with optimal conventional therapy and were hospitalized for administration of intravenous medications as needed. RESULTS: The baseline characteristics of groups 1 versus 2, including New York Heart Association functional class (IV in both groups), admission systolic arterial blood pressure (99 +/- 14 vs. 97 +/- 13 mm Hg), right atrial pressure (13 +/- 6 vs. 14 +/- 6 mm Hg), pulmonary capillary wedge pressure (28 +/- 7 vs. 31 +/- 10 mm Hg), serum sodium (136 +/- 7 vs. 139 +/- 6 mEq/L) and serum creatinine (1.7 +/ 0.8 vs. 1.8 +/- 1.8 mg/dL), were similar. The 6-month (51% vs. 18%) and 1-year (36% vs. 9%) survival rates were significantly higher (P = 0.001 for both) in group 1 than in group 2. In addition, patients treated with intermittent intravenous inotropic agents improved their functional and hemodynamic status. CONCLUSIONS: Intermittent intravenous inotropic agents combined with prophylactic oral amiodarone seem to improve the outcomes of patients with end-stage chronic heart failure. Further research is warranted to elucidate whether this treatment strategy should be considered as a standard therapy in patients with refractory end-stage heart failure. PMID- 19188833 TI - Effect of oxidized low-density lipoprotein on survival and function of endothelial progenitor cell mediated by p38 signal pathway. AB - This study was designed to investigate whether oxidized low-density lipoprotein (oxLDL) affects the survival and activity of endothelial progenitor cell (EPC) mediated by p38 mitogen-activated protein kinase (MAPK). EPCs were isolated from human peripheral blood in endothelial cell growth medium-2. Incubation with oxLDL at 100 microg/mL decreased EPC number. Treated with oxLDL resulted in increase of EPC apoptosis and in decrease of EPC proliferation. Treatment with oxLDL resulted in a significantly reduced migratory rate of EPCs and reduced adhesion to fibronectin. Treatment with oxLDL impaired the in vitro angiogenesis ability of EPCs. However, all the detrimental effects on EPC were attenuated by pretreatment of EPCs with SB203580, an inhibitor of the p38 MAPK. In addition, the inhibition of the p38-kinase by SB203580 also significantly improved basal number and functions of EPCs. Western blot analysis revealed that oxLDL induced dose- and time-dependent activation of the p38 MAPK. These results demonstrated that p38 MAPK plays a critical role in regulating the number and functions of EPCs in vitro. SB203580 can improve the number and functions of EPCs under basal conditions and prevent the negative effects of oxLDL on the number and functions of EPCs and may be useful to improve the number and function of EPCs for potential cell therapy. PMID- 19188834 TI - Effect of 3-thienylalanine-ornithine-proline, new sulfur-containing angiotensin converting enzyme inhibitor on blood pressure and oxidative stress in spontaneously hypertensive rats. AB - Sulfur-containing angiotensin-converting enzyme (ACE) inhibitors have an edge over other inhibitors in improving endothelial dysfunction and reducing oxidative stress. In this study, effect of new sulfur-containing ACE inhibitor, 3 thienylalanine-ornithine-proline (TOP), was studied on blood pressure (BP) and oxidative stress in Spontaneously Hypertensive Rats (SHRs). Acute oral administration of 5 mg/kg of TOP significantly lowered systolic blood pressure for longer periods (18 +/- 0.5 hours, P < 0.05 vs. 16 +/- 0.5 hours) than captopril (20 mg/kg). Thiophene ring of TOP is devoid of free sulfur and may avoid early oxidation resulting in longer duration of action. Chronic oral administration of TOP (5 mg/kg twice a day) for 7 days lowered BP from 189.1 +/- 2.5 to 161.7 +/- 2.6 mm Hg (P < 0.05), decreased the ACE activity, and increased the nitrite levels in various tissues (P < 0.05) and serum (P 0.05). In the multivariate analysis, incontinence (prevalence odds ratio [POR], 1.67; 95% CI, 1.02-2.73), vaginal dryness (POR, 3.97; 95% CI, 2.49-6.31), vaginal itching (POR, 2.44; 95% CI, 1.55-3.83), loss of libido (POR, 3.08; 95% CI, 1.92-4.94), and partnership (POR, 2.22; 95% CI, 1.29-3.82) remained associated with dyspareunia. CONCLUSIONS: Our results agree with previous studies regarding the potential association between health and lifestyle factors and dyspareunia. Additional studies of female sexual dysfunction in Puerto Rican women are highly warranted. PMID- 19188848 TI - Exercise program affects body composition but not weight in postmenopausal women. AB - The aim of this study was to investigate the effect of a 12-month moderate-to vigorous exercise program combining aerobic and muscle strength training on body composition among sedentary, postmenopausal women. METHODS: A randomized controlled trial was conducted. A total of 189 sedentary postmenopausal women (age 50-69 y, body mass index 22-40 kg/m2) were randomly assigned to an exercise (n = 96) or a control group (n = 93). Study parameters measured at baseline, 4 months, and 12 months were as follows: body weight and body height (body mass index), waist and hip circumference (body fat distribution), and dual-energy x ray absorptiometry (total body fat and lean mass). Differences in changes in study parameters between exercise and control group were examined with generalized estimating equations analysis. RESULTS: The exercise program did not result in significant effects on weight, body mass index, and hip circumference. The exercise group experienced a statistically significant greater loss in total body fat, both absolute (-0.33 kg) (borderline) as in a percentage (-0.43%) compared with the control group. In addition, lean mass increased significantly (+0.31 kg), whereas waist circumference (-0.57 cm) decreased significantly compared with the control group. CONCLUSIONS: We conclude that a 12-month exercise program combining aerobic and muscle strength training did not affect weight but positively influenced body composition of postmenopausal women. Affecting body fat distribution and waist circumference may have important health implications because it is an independent risk factor in obese but also in nonobese people. Therefore, this study gives further credence to efforts of public health and general practitioners aiming to increase physical activity levels of postmenopausal women. PMID- 19188849 TI - Temporal associations of hot flashes and depression in the transition to menopause. AB - OBJECTIVE: The aim of this study was to evaluate associations between hot flashes and depressed mood in the menopausal transition and associations of these symptoms with reproductive hormone changes. METHODS: A 10-year follow-up in a population-based cohort of women who had no experience of hot flashes or depressed mood at baseline was conducted. RESULTS: The incidence of hot flashes significantly increased compared with the incidence of depressed mood in the 10 year follow-up (P < 0.001). Sixty-seven percent of the women reported hot flashes, 50% reported depressed mood, and 41% reported both symptoms during the study interval. Reporting of both hot flashes and depressed mood was greater than expected if the processes operated independently (P < 0.001). Of the women who experienced both symptoms, depressed mood was more likely to precede hot flashes (relative risk = 2.1; 95% CI, 1.5-2.9). Within-woman increases in follicle stimulating hormone levels were associated with the onset of depressed mood in unadjusted analysis (P = 0.05). Increased follicle-stimulating hormone levels, decreased inhibin B levels, and the variability of estradiol were significantly associated with hot flashes. Follicle-stimulating hormone and inhibin B remained significantly associated with hot flashes in the final multivariable models (P < 0.001). CONCLUSIONS: Both hot flashes and depressive symptoms occur early in the menopausal transition in women with no previous experience of these symptoms. Depressive symptoms are more likely to precede hot flashes in women who report both symptoms. The findings support the concept that the changing hormonal milieu of the menopausal transition is one of multiple factors associated with the onset of symptoms. PMID- 19188850 TI - Efficacy of nonestrogenic hot flash therapies among women stratified by breast cancer history and tamoxifen use: a pooled analysis. AB - OBJECTIVE: Various nonestrogenic therapies have been found to be effective in mitigating hot flashes, but it has been unclear whether the efficacy varies by whether women have had breast cancer and/or were taking tamoxifen. METHODS: This study used data from Mayo Clinic/North Central Cancer Treatment Group clinical trials that evaluated the efficacy of any nonestrogenic agent for hot flashes and had information on breast cancer history or tamoxifen use. Statistically significant changes from the fourth treatment week versus the baseline week, using individual patient data, were assessed using Student's t test. RESULTS: A total of 1,396 women from 20 hot flash studies were eligible for analysis. Overall, women without breast cancer had a similar percentage of baseline hot flash score at week 4, as did those with breast cancer (53% vs 50%, P = 0.92). Women who were not taking tamoxifen had a significantly lower percentage of hot flash score at week 4 as compared with those who used tamoxifen (54% vs 61%, P = 0.01). However, this was due to a higher reduction in hot flash scores in the placebo arms among women not receiving tamoxifen; the percentage reduction in hot flash scores at week 4 from baseline in the active therapy arms of the randomized placebo-controlled trials (ie, excluding placebo arms) was similar among the tamoxifen users and nonusers (difference in mean percentage reduction, 5.7; 95% CI, -1.76 to 13.16). CONCLUSIONS: Some nonestrogenic therapies seem to be useful for reducing hot flashes, irrespective of the etiology of hot flashes. PMID- 19188851 TI - Serum antimullerian hormone predicts ovarian reserve in a monkey model. AB - OBJECTIVE: A monkey model of the menopausal transition (perimenopause) would facilitate efforts to understand better the effect of hormonal fluctuations during this life phase on the initiation of chronic diseases associated with the postmenopausal years. Antimullerian hormone (AMH) is a promising marker of ovarian reserve (primordial follicle number) in women. Here, we describe the relationship between AMH and ovarian reserve in cynomolgus monkeys (Macaca fascicularis) estimated to be 12 to 15 years of age (approximately 36-45 y in women). METHODS AND RESULTS: The results of daily vaginal swabbing (to detect menses) and thrice weekly blood sampling for 12 weeks indicate that AMH is relatively stable across the menstrual cycle (intraclass correlation, approximately 0.80), with a slight although significant (P < 0.02) reduction (approximately 1.4-fold) on days 2 to 5 postovulation. Substantial interindividual variation in AMH concentrations were observed between monkeys, with values ranging from 4.46 +/- 0.17 to 18.80 +/- 0.71 ng/mL (mean +/- SE). Antimullerian hormone concentrations were reduced by approximately 63% after the removal of one ovary (7.6 +/- 0.77 vs 2.75 +/- 0.37 ng/mL; P < 0.001; n = 19) and were below the level of detection after the removal of both ovaries (5.8 +/- 0.42 to <0.05 ng/mL; P < 0.001; n = 84), suggesting that the ovary is likely to be either the major or the sole source of AMH in the monkey. Finally, we examined the association between AMH and primordial, primary, and secondary follicles in 29 monkeys and found significant associations with all follicle types (r = 0.78, r = 0.66, and r = 0.80, respectively; P < 0.01). CONCLUSIONS: The relationship between AMH and ovarian reserve in the monkey is similar to that in women, suggesting that monkeys may be a useful model for studying hormonal fluctuations across the menopausal transition. PMID- 19188852 TI - Duration of vasomotor symptoms in middle-aged women: a longitudinal study. AB - OBJECTIVE: Vasomotor symptoms adversely affect the quality of life and functional status of most women during the menopausal transition, but little is known about how long these symptoms last. The most effective treatment, hormone therapy (HT), carries risks and benefits that depend on the timing and duration of use. In this study we sought to estimate the duration of vasomotor symptoms in a longitudinal study. METHODS: We reanalyzed primary data from 438 women in the longitudinal cohort of the population-based Melbourne Women's Midlife Health Project. Two hundred and five women who had completed 13 years of follow-up were included in the analyses. The onset and cessation of vasomotor symptoms were reported, stratifying analyses according to ever use of HT. Symptom duration was calculated as the time between the first and last bothersome hot flush reported. RESULTS: The mean (SD) duration of bothersome menopausal symptoms for women who completed 13 years of follow-up and who never used HT was estimated to be 5.2 (3.8) years (median, 4 years). If women who used HT were included, the mean (SD) duration was 5.5 (4.0) years (median, 4 years). The estimated duration of symptoms varied according to the duration of longitudinal follow-up, with a mean estimate of 3.4 years (median, 3 years) when only 8 years of follow-up data were analyzed. The only factor associated with duration of hot flushes was regular exercise-more exercise was associated with shorter symptom duration. CONCLUSIONS: The average duration of vasomotor symptoms in this sample is more than 5 years, substantially longer than had been previously reported. PMID- 19188853 TI - Is there an association between vasomotor symptoms and both low bone density and cardiovascular risk? PMID- 19188855 TI - Replacing hormone therapy--is the decline in prescribing sustained, and are nonhormonal drugs substituted? AB - OBJECTIVES: After two cautioning landmark studies in 2002 and 2003, a dramatic decrease in hormonal therapy (HT) prescribing for menopausal symptoms was seen. Our objectives were to (1) determine whether this decline in HT prescribing sustained until 2007 and (2) investigate nonhormonal drug prescribing for women who stopped HT. METHODS: We analyzed drug dispensing data from community pharmacies in The Netherlands. For the first objective, we analyzed exposure prevalence of HT. For our second objective, we selected all women who were dispensed HT prescriptions between August 2002 and August 2003. From this study cohort, we defined our index group as all women who stopped HT shortly after the studies (n = 1,254) and a reference group as all women who continued HT (n = 839). We calculated the incidence of nonhormonal therapies for both groups, reporting risk ratio (RR) and 95% CI. Kaplan-Meier curves were also constructed. RESULTS: Mean exposure prevalence of HT (per 1,000 women) pre 2002 versus post 2004 was 30.6 versus 15.3 (50.0% decline) for 40 to 49 years, 79.2 versus 25.5 (67.7% decline) for 50 to 59 years, and 28.4 versus 11.6 (59.1% decline) for 60 to 69 years. HT exposure remained low until 2007. HT stoppers receive more clonidine, RR 3.48 (2.36-5.13); anxiolytics or sedatives, RR 1.46 (1.15-1.87); and osteoporosis prophylaxis and treatment, RR 2.04 (1.14-3.66). Young stoppers (40-49 y) received more antidepressants, RR 2.70 (1.41-5.11), whereas older stoppers (60-69 y) received less antidepressants, RR 0.43 (0.18-1.05). Kaplan Meier curves showed that nonhormonal drug prescribing occurred soon after HT was stopped. CONCLUSIONS: This study shows the dramatic and sustained impact of the cautioning landmark studies on HT prescribing. HT stoppers received more nonhormonal therapies for menopausal symptoms compared with those who continued HT. PMID- 19188856 TI - Impaired theta phase-resetting underlying auditory N1 suppression in chronic alcoholism. AB - It has been suggested that chronic alcoholism may lead to altered neural mechanisms related to inhibitory processes. Here, we studied auditory N1 suppression phenomena (i.e. amplitude reduction with repetitive stimuli) in chronic alcoholic patients as an early-stage information-processing brain function involving inhibition by the analysis of the N1 event-related potential and time-frequency computation (spectral power and phase-resetting). Our results showed enhanced neural theta oscillatory phase-resetting underlying N1 generation in suppressed N1 event-related potential. The present findings suggest that chronic alcoholism alters neural oscillatory synchrony dynamics at very early stages of information processing. PMID- 19188857 TI - The effects of emotional intensity on ERP correlates of recognition memory. AB - The effects of negative emotional intensity on memory-related brain activity were tested by using human scalp event-related potentials (ERP). A neural index of memory function--the electrophysiological 'Old-New' effect--was obtained from participants undertaking a memory recognition test of previously studied ('old') and unstudied ('new') pictures of variable levels of negative emotional intensity. The magnitude of the old-new effect was compared across four different levels of linearly increasing stimulus emotional intensity. Results revealed an inverted-U-shaped effect of emotional intensity on the magnitude of ERP old-new differences starting at 300 ms after stimulus onset. These results suggest that moderate negative emotions can enhance memory brain function, whereas extreme levels of emotional intensity have the potential of inhibiting memory function. Results are discussed in terms of their implications for neurobiological and psychological models of emotion-memory interactions. PMID- 19188858 TI - L-dopa-induced desensitization depends on 5-hydroxytryptamine imbalance in hemiparkinsonian rats. AB - We have shown before that 2-week intrastriatal L-3,4-dihydroxyphenylalanine (L dopa) infusion significantly decreased contralateral rotations induced by acute intraperitoneal L-dopa/carbidopa and increased striatal tryptophan hydroxylase in 6-hydroxydopamine-lesioned rats. Here, we examined the effect of acutely administered L-dopa (10 microg) into 6-hydroxydopamine-lesioned rat striata under the inhibition of tryptophan hydroxylase by 4-chloro-DL-phenylalanine. Acute intrastriatal L-dopa infusion significantly decreased contralateral rotations induced by intraperitoneal L-dopa/carbidopa (10/30 mg/kg) 1 and 7 days after intrastriatal L-dopa. This desensitization to L-dopa occurred only when there was a striatal 5-hydroxytryptamine (5-HT) imbalance, not when 5-HT levels in the intact and lesioned sides were similar, either very low (day 1 postinfusion) or similarly recovered (day 7 postinfusion). We conclude that 5-HT plays a significant role in the striatal dopaminergic imbalance that evokes the rotational behavior. PMID- 19188860 TI - Sonic hedgehog signaling in the chick retina accelerates Meis2 downregulation simultaneously with retinal ganglion cell genesis. AB - The spatial-temporal expression and function of Drosophila melanogaster homothorax (hth) and its vertebrate homologue Meis2 in the eye anlage show a high degree of evolutionary conservation. Both are expressed by progenitor cells ahead of the neurogenic wave front, promote rapid proliferation of these cells, and are downregulated before cells exit the cell cycle and differentiate. Here, we show that downregulation of Meis2 accompanies, but is not required, for retinal differentiation. In addition, we provide evidence that the mechanisms that terminate expression of both genes differ. Hth expression in the fly eye imaginal disc is primarily repressed by the transforming growth factor beta family protein decapentaplegic, whereas Meis2 expression in the chick eye cup is terminated in response to sonic hedgehog signaling. PMID- 19188859 TI - In-vitro effects of brain-derived neurotrophic factor on neural progenitor/stem cells from rat hippocampus. AB - Mounting evidences from in-vivo studies showed that brain-derived neurotrophic factor (BDNF) plays an important role in the neurogenesis, but the effects of BDNF are controversial because of neurogenesis affected by many factors in vivo. In this study, we investigated the effects of BDNF on the survival, proliferation, and differentiation of the neural progenitor/stem cells (NPCs) in vitro. The results showed that 40 ng/ml of BDNF significantly increased the number and diameter of neurospheres formed by NPCs; meanwhile, the TUNEL rates and lactate dehydrogenase release of NPCs were also inhibited. Tuj-1+ immunostaining showed that BDNF obviously induced the NPCs to differentiate into neurons and elongated the neurite. Our results implied that BDNF promotes the proliferation of NPCs and induces them to differentiate into neurons, and enhancement of the survival of NPCs probably is one of the mechanisms. PMID- 19188861 TI - N1 component reflects difference of terminal chords in three-chord sequences. AB - We understand from experience that musical contexts are formed when chords are combined according to the rules of harmony. In this study, the N1 component of the auditory-evoked potential was measured using comparable three-chord sequences; these sequences were constructed as a consecutive task (cf. C-C-C vs. Cm-Cm-Cm; control) and a cadence task (cf. C-G-C vs. C-G-Cm). In the cadence task, compared with cadences ending with a major chord (anticipated chord), those ending with a minor chord (unanticipated chord) showed a significantly larger amplitude of N1 waves. These components of auditory-evoked potentials reflect the effect of chord progression in musical perception and suggest that the musical context is recognized at least 100 ms after a chord is played. PMID- 19188862 TI - Lateralized sex differences in stress-induced dopamine release in the rat. AB - This study examined the possibility that hemispheric differences in stress induced brain activation vary as a function of sex. Using in-vivo voltammetry, increases in extracellular dopamine release in response to predator odour and tail pinch stress were recorded bilaterally and simultaneously in either the infralimbic cortex or basolateral amygdala. In both stress-sensitive brain regions, significant sex x hemisphere interactions were observed, with males and females showing greater dopamine activation in right-brain and left-brain structures, respectively. Cortical asymmetries in dopamine release also showed sex-specific correlations with stress-induced neuroendocrine activation. Given the intriguing human parallels, we suggest that differential cerebral lateralization may be highly relevant to the disproportionately high incidence of stress-related disorders such as depression and anxiety seen in women. PMID- 19188863 TI - Induction of antioxidant enzymes by curcumin and its analogues in human islets: implications in transplantation. AB - OBJECTIVES: The survival of transplanted human islets is hampered by the quality of islets, which is affected by oxidative stress during isolation. The objective of this study was to determine if curcumin and its analogues could induce antioxidant enzymes in beta cells of human islets. METHODS: The expression of antioxidant enzymes in isolated human islets exposed to curcuminoids was determined at the messenger RNA levels by real-time quantitative reverse transcription-polymerase chain reaction using Taqman probes and at the protein level by Western blot analysis. Double immunofluorescent staining of islets was carried out to determine the induction of antioxidant enzymes in beta cells. RESULTS: Curcuminoids induced the expression of heme oxygenase 1; modulatory subunit of gamma-glutamyl-cysteine ligase; and NAD(P)H:quinone oxidoreductase 1 at the messenger RNA levels by 2- to 12-fold and at the protein levels by 2- to 6 fold in human islets. Increased expression of antioxidant enzymes was seen in beta cells of islets as shown by immunofluorescent staining. Curcuminoids also increased the islet content of glutathione (a product of the modulatory subunit of gamma-glutamyl-cysteine ligase) and the basal insulin secretion and protected them from oxidative stress. CONCLUSIONS: Our observations suggest that curcumin or its analogues could be used to induce cellular defense against oxidative stress and improve islet transplantation outcomes. PMID- 19188864 TI - Inhibition of transforming growth factor beta signaling by halofuginone as a modality for pancreas fibrosis prevention. AB - OBJECTIVES: Chronic pancreatitis is characterized by inflammation and fibrosis. We evaluated the efficacy of halofuginone, an inhibitor of collagen synthesis and myofibroblast activation, in preventing cerulein-induced pancreas fibrosis. METHODS: Collagen synthesis was evaluated by in situ hybridization and staining. Levels of prolyl 4-hydroxylase beta (P4Hbeta), cytoglobin/stellate cell activation-associated protein (Cygb/STAP), transgelin, tissue inhibitors of metalloproteinases, serum response factor, transforming growth factor beta (TGFbeta), Smad3, and pancreatitis-associated protein 1 (PAP-1) were determined by immunohistochemistry. Metalloproteinase activity was evaluated by zymography. RESULTS: Halofuginone prevented cerulein-dependent increase in collagen synthesis, collagen cross-linking enzyme P4Hbeta, Cygb/STAP, and tissue inhibitors of metalloproteinase 2. Halofuginone did not affect TGFbeta levels in cerulein-treated mice but inhibited serum response factor synthesis and Smad3 phosphorylation. In culture, halofuginone inhibited pancreatic stellate cell (PSC) proliferation and TGFbeta-dependent increase in Cygb/STAP and transgelin synthesis and metalloproteinase 2 activity. Halofuginone increased c-Jun N terminal kinase phosphorylation in PSCs derived from cerulein-treated mice. Halofuginone prevented the increase in acinar cell proliferation and further increased the cerulein-dependent PAP-1 synthesis. CONCLUSIONS: Halofuginone inhibits Smad3 phosphorylation and increases c-Jun N-terminal kinase phosphorylation, leading to the inhibition of PSC activation and consequent prevention of fibrosis. Halofuginone increased the synthesis of PAP-1, which further reduces pancreas fibrosis. Thus, halofuginone might serve as a novel therapy for pancreas fibrosis. PMID- 19188865 TI - Serine protease inhibitor 8 is a novel immunohistochemical marker for neuroendocrine tumors of the pancreas. AB - OBJECTIVES/METHODS: The intracellular serine protease inhibitor 8 (SERPINB8) is expressed by squamous epithelium, monocytes, and a subset of neuroendocrine cells. Using immunohistochemistry, we now have further investigated the expression of SERPINB8 in normal neuroendocrine cells and its potential use as a marker to identify neuroendocrine tumors of the pancreas. RESULTS: In normal neuroendocrine tissues, strongest SERPINB8 expression was detected in islets of Langerhans of the pancreas. Moderate SERPINB8 expression was observed in neuroendocrine cells of the thyroid, adrenal cortex, colon, and pituitary gland. Fluorescent double staining revealed that in the pancreas, SERPINB8 is specifically expressed by insulin-producing beta cells. In a panel of 20 patients with pancreatic islet cell tumors, however, SERPINB8 was broadly expressed and not restricted to insulinomas. In islet cell tumors, SERPINB8 had a similar diagnostic sensitivity as compared with the widely used neuroendocrine markers chromogranin A and synaptophysin. When SERPINB8 was combined with these 2 markers, an even higher diagnostic sensitivity was reached. In contrast, exocrine adenocarcinomas of the pancreas showed no SERPINB8 expression. CONCLUSIONS: The SERPINB8 is expressed in normal neuroendocrine cells of several organs as well as in neuroendocrine tumors of the pancreas, where it can be used as an additional diagnostic immunohistochemical marker. PMID- 19188866 TI - Physical compatibility of neonatal total parenteral nutrition admixtures containing organic calcium and inorganic phosphate salts in a simulated infusion at 37 degrees C. AB - OBJECTIVE: Within an incubator environment at 37 degrees C, we evaluated the physical compatibility of calcium (Ca) and phosphate (P) in simulated neonatal total parenteral nutrition (TPN) infusions. DESIGN: Representative neonatal TPN formulas containing four concentrations of amino acids [AA] (1-4 g/dL) and two levels of dextrose [D] (5 and 10 g/dL) were monitored for calcium-phosphate precipitate formation over 24 hours using a laser-based particle counting technique. SETTING: Research Laboratory. SUBJECTS: None. MEASUREMENTS AND MAIN RESULTS: Of the eight AA-D combinations studied, two formulas containing 1 g/dL AA with either 5 g/dL or 10 g/dL of D and high amounts of Ca and P resulted in significant increases in particle counts and visibly evident precipitation during the 24-hour infusion. Precipitation was noted only after the infusion passed through the incubator, but not in the TPN bags, demonstrating the heightened risk of calcium-phosphate incompatibility with increased temperatures. CONCLUSIONS: Low AA concentrations and high temperatures are significant risk factors for the precipitation of the insoluble dibasic calcium phosphate that may be fatal upon intravenous infusion. When thermoregulation is required, the AA concentration of the TPN formula should not be less than 1.5 g/dL (and ideally > or =3 g/dL) when high intakes of calcium (60 mg/dL) and phosphate (46.5 mg/dL) are prescribed. PMID- 19188867 TI - Efficacy of sedation regimens to facilitate mechanical ventilation in the pediatric intensive care unit: a systematic review. AB - OBJECTIVE: Children admitted to pediatric intensive care units (PICUs) often receive sedatives to facilitate mechanical ventilation. However, despite their widespread use, data supporting appropriate dosing, safety, and optimal regimens for sedation during mechanical ventilation are lacking. Therefore, we conducted a systematic review of published data regarding efficacy of sedation to facilitate mechanical ventilation in PICU patients. Our primary objective was to identify and evaluate the quality of evidence supporting sedatives used in PICUs for this purpose. DATA SOURCES: We searched MEDLINE, EMBASE, and The Cochrane Registry of Clinical Trials from 1966 to June 2008 to identify published articles evaluating sedation regimens to facilitate mechanical ventilation in PICU patients. STUDY SELECTION: We included only those studies of intubated PICU or pediatric cardiac intensive care unit patients receiving pharmacologic agents to facilitate mechanical ventilation that reported quality of sedation as an outcome. DATA EXTRACTION: We analyzed studies separately for study type and by agents being studied. Studies were appraised using criteria of particular importance for reviews evaluating sedatives. DATA SYNTHESIS: Our search strategy yielded 39 studies, including 3 randomized trials, 15 cohort studies, and 21 cases series or reports. The 39 studies evaluated a total of 39 different sedation regimens, with 21 different scoring systems, in a total of 901 PICU/cardiac intensive care unit patients ranging in age from 3 days to 19 years old. Most of the studies were small (<30 patients), and only four studies compared one or more agents to another. Few studies thoroughly evaluated drug safety, and only one study met all quality criteria. CONCLUSIONS: Despite the widespread use of sedatives to facilitate mechanical ventilation in the PICU, we found that high-quality evidence to guide clinical practice is still limited. Pediatric randomized, controlled trials with reproducible methods and assessment of drug safety are needed. PMID- 19188868 TI - Critical appraisal of: Koeman M, van der Ven AJ, Hak E, et al: Oral decontamination with chlorhexidine reduces the incidence of ventilator-associated pneumonia. Am J Respir Crit Care Med 2006; 173:1348-1355. PMID- 19188869 TI - Dexmedetomidine sedation for pediatric post-Fontan procedure patients. AB - OBJECTIVE: The hemodynamic, respiratory, and sedative effects of dexmedetomidine (DEX) for pediatric patients post-Fontan surgery. DESIGN: Retrospective. SETTING: Single institutional intensive care unit. PARTICIPANTS: Fourteen patients undergoing Fontan-type surgery. RESULT: A retrospective review was conducted on 14 pediatric patients who had undergone a Fontan procedure for congenital heart disease. A vital component of postoperative management of these patients is to prevent an increase in pulmonary vascular resistance (PVR) that may lead to a serious reduction in cardiac output. DEX an alpha-2 adrenergic receptor agonist might offer an advantage over current sedation methods in preventing a rise in PVR. Nine patients received sedation with DEX and five patients in a control group were administered standard regimens of sedation and analgesia. The DEX group exhibited no evidence of an increased partial pressure of arterial carbon dioxide postoperatively as opposed to the control group. This lack of respiratory depression made the DEX group less likely to increase their PVR. However, the DEX group did experience a significant incidence of bradycardia that required the use of a cardiac pacemaker. CONCLUSIONS: The results of this retrospective review of the role of DEX in the management of the post-Fontan surgical pediatric patient indicate some potential advantages. PMID- 19188870 TI - Exposure to the pharmaceutical excipients benzyl alcohol and propylene glycol among critically ill neonates. AB - OBJECTIVE: To document neonatal exposures to the potentially harmful pharmaceutical excipients benzyl alcohol (BA) and propylene glycol (PG) present in parenteral medications routinely administered in the intensive care unit. DESIGN: Retrospective, observational study. SETTING: Neonatal and pediatric intensive care units of a tertiary care, university hospital. PATIENTS: Randomly selected sample of 170 episodes of exposure to parenteral medications containing BA (n = 88) or PG (n = 82). MEASUREMENTS: We identified all medication sources of BA or PG administered to study neonates during hospitalization, and calculated cumulative doses (mg/kg/day and mg/day) of BA or PG received as a result of exposure to those medications. MAIN RESULTS: We observed a wide range in the cumulative excipient dose received by neonates. Median (range) cumulative dose was 4.5 mg/kg/day (0.6-319.5 mg/kg/day) for BA, and 204.9 mg/kg/day (17.3-9472.7 mg/kg/day) for PG. Patients who received medications via continuous infusion received significantly higher excipient doses than patients who received medications intermittently (p < 0.0001). In this subset of patients, median cumulative excipient doses (BA, 106.3 mg/kg/day and PG, 4554.5 mg/kg/day) were approximately 21 and 180 times the acceptable daily intakes of BA and PG (5 and 25 mg/kg/day), respectively, and exceeded the doses above which toxicity has been reported in infants. No significant correlation between duration of medication administration and cumulative excipient exposure was identified for BA or PG. Midazolam and lorazepam were involved in over two-thirds of BA and PG exposures, respectively. CONCLUSIONS: Critically ill neonates, especially those receiving medications by continuous infusion, are at risk of being exposed to BA and PG at potentially toxic doses during routine medication administration. Given the serious adverse reactions known to be associated with BA and PG, future studies are warranted to determine the clinical consequences associated with this degree of excipient exposure. PMID- 19188871 TI - Nosocomial infections in pediatric cardiovascular surgery patients: a 4-year survey. AB - OBJECTIVE: To determine the prevalence, characteristics, and risk factors for nosocomial infections (NIs) in children postcardiac surgery and hospitalized in a pediatric intensive care unit (PICU). DESIGN: Case control study. SETTING: PICU of a tertiary care university-affiliated medical center. PATIENTS AND CONTROLS: Between January 1, 1999 and December 31, 2002, 356 children underwent cardiac surgery and were admitted to the PICU (381 admissions). There were 146 episodes (91 patients) of NI according to the Centers for Disease Control and Prevention criteria. The control group (92 patients) was drawn as a random sample from admissions with no evidence of NI. MEASUREMENTS AND MAIN RESULTS: Data retrieved from medical records included demographic information, type of operation, complexity score, extrinsic risk factors (invasive devices, postoperative complications, etc.), specific pathogens, therapeutic interventions, and outcome. There were 146 episodes of NI during 381 admissions, yielding a nosocomial infection ratio of 38.3%, and a prevalence of PICU-acquired infection of 24.4% (93 admissions with NI out of a total of 381 admissions). The most common NI sites were the bloodstream and the lower respiratory tract (65.8% and 16.4%, respectively). The main causative organisms were Coagulase negative Staphylococcus, Klebsiella pneumonia, and Candida spp. (18.8%, 16.7%, and 15%, respectively). Multivariate analysis revealed the following risk factors for NI: age < 2 months, congenital malformations, post operative complications, and open chest procedure. The crude mortality rate for patients with NI was 23.7%, compared with 2.2% for patients without NI (p < 0.001). Predictors for mortality in patients with NI were post operative complications, open-chest procedure, sepsis, and urinary tract infection. CONCLUSIONS: Our results draw attention to the importance of NI and their influence on survival in pediatric patients undergoing cardiac surgery. Prevention and control measures may reduce these infections and subsequently reduce morbidity and mortality in this vulnerable population. PMID- 19188872 TI - Derivation and validation of an equation for adjustment of neuron-specific enolase concentrations in hemolyzed serum. AB - OBJECTIVE: To derive and validate a formula to allow for adjustment of serum neuron-specific enolase (NSE) concentrations based on the amount of hemolysis in the sample. To compare the accuracy of qualitative and quantitative assessment of hemolysis. DESIGN: Derivation and validation of a formula. SUBJECTS: Eighty-three children <11 years of age were used for the derivation. Twenty-three children <11 years without brain injury and 30 with brain injury were used for the validation. INTERVENTIONS: We evaluated the amount of hemolysis quantitatively using a hemocue and qualitatively by visualization. We measured NSE concentrations by enzyme-linked immunosorbent assay. Using a subset of subjects, we used linear regression analysis to derive an equation to adjust serum NSE concentrations based on the degree of hemolysis. We prospectively validated the formula in a different group. NSE < or =11.5 ng/mL was considered normal. RESULTS: The formula derived was as follow: Adjusted NSE = Unadjusted NSE - (1.659) (square root of the hemocue). In the validation, 96% of children without brain injury had a normal adjusted NSE; 93% of children with brain injury had an abnormal adjusted NSE. Qualitative analysis of hemolysis was inconsistent and both overassessed and under-assessed hemolysis as quantified by hemocue. CONCLUSIONS: We retrospectively derived and prospectively validated an equation for adjusting serum NSE concentrations based on the amount of hemolysis in the sample. Use of this formula will allow for accurate measurement of NSE even in hemolyzed sample and may improve its usefulness as a marker of brain injury in children. Qualitative assessment of the degree of hemolysis is not accurate and should not be used. PMID- 19188873 TI - Vasopressin for in-hospital pediatric cardiac arrest: results from the American Heart Association National Registry of Cardiopulmonary Resuscitation. AB - OBJECTIVE: To describe the landscape of vasopressin uses reported to the American Heart Association National Registry of cardiopulmonary resuscitation (CPR) and test the hypothesis that vasopressin use will be associated with improved return of a sustained circulation (ROSC) following in-hospital pediatric cardiac arrest. DESIGN: Multicentered, national registry of in-hospital CPR. SETTING: One hundred seventy-six North American Hospitals reporting to registry from October 1999 to November 2004. PATIENTS: Totally, 1293 consecutive pediatric patients with pulseless cardiac arrest meeting criteria for analysis identified from a registry of all patients resuscitated for cardiac arrest. Inclusion criteria were age <18 years, chest compressions and/or defibrillation, in-hospital location, and documented resuscitation record. INTERVENTIONS: None. MEASUREMENTS AND OUTCOMES: Prearrest, event, cardiopulmonary resuscitation, and postresuscitation variables were collected. Primary outcome variable was ROSC >20 minutes. Secondary survival outcomes included 24 hour, discharge and favorable neurologic survival on hospital discharge. Descriptive, univariate, and multivariable analysis to evaluate the association of vasopressin with survival outcomes were performed. RESULTS: Only 5% of patients received vasopressin in this review. Vasopressin was most often given in a pediatric hospital (57%) and in and intensive care setting (76.6%). Patients who were given vasopressin had longer arrest duration (median 37 minutes) vs. those who did not (24 minutes) (p = 0.004). In multivariate analysis, vasopressin was associated with worse ROSC but no difference in 24 hours or discharge survival. CONCLUSION: Vasopressin was given infrequently in in hospital cardiac arrest. It was most likely to be given in an intensive care setting, and in a pediatric hospital. Multivariate analysis shows an association with vasopressin use and worse ROSC. PMID- 19188874 TI - Successful placement of postpyloric enteral tubes using electromagnetic guidance in critically ill children. AB - OBJECTIVES: Initiation of postpyloric feeding is often delayed by difficulties in placement of enteral tubes. We evaluated the effectiveness of bedside postpyloric enteral tube (PET) placement using an electromagnetic (EM)-guided device. We hypothesized that: 1) EM-guided placement of PETs would be successful more often than standard blind placement with a shorter total time to successful placement and 2) the EM-guided technique would have similar overall costs to the standard technique. DESIGN: Prospective cohort trial with serial control groups in a pediatric intensive care unit at a tertiary care children's hospital. INTERVENTIONS: We collected data on a cohort of consecutive pediatric intensive care unit patients who underwent PET placement by standard blind technique followed by a cohort who underwent EM-guided placement. The primary outcome measure was successful placement determined by abdominal radiography. MEASUREMENTS AND MAIN RESULTS: One hundred seven patients were evaluated in the trial: 57 in the standard group and 50 in the EM-guided group. Demographic data, percent intubated, and admission diagnosis were similar in both groups. Forty-one of 50 patients (82%) in the EM-guided group had successful placement compared with 22 of 57 in the standard group (38%) (p < 0.0001). The average time to successful placement was 1.7 vs. 21 hours in the EM-guided group and standard group, respectively (p < 0.0001). Children in the EM-guided group received fewer radiographs (p = 0.007) and were given more prokinetic drugs (p = 0.045). There were no episodes of pneumothorax in either group. After controlling for prokinetic drug use, EM-guided placement was more likely to result in successful placement than the standard blind technique (odds ratio 6.4, 95% confidence interval 2.5-16.3). An annual placement rate of 250 PETs by EM guidance, based on our institution's current utilization rates, is associated with a cost savings of $55.46 per PET placed. CONCLUSION: EM guidance is an efficient and cost-effective method of bedside PET placement. PMID- 19188875 TI - The effect of aprotinin, tranexamic acid, and aminocaproic acid on blood loss and use of blood products in major pediatric surgery: a meta-analysis. AB - OBJECTIVE: Aprotinin reduces the blood loss and transfusion of blood products in children undergoing major surgery. Aprotinin has been associated with severe side effects in adults, and tranexamic acid and aminocaproic acid have been found to be safer alternatives in adults. This systematic review addresses the question of whether tranexamic acid and aminocaproic acid are equally effective as aprotinin for reducing blood loss and transfusion in children undergoing major surgery. DATA SOURCES: A systematic review of the literature was conducted to identify all randomized controlled trials of aprotinin, tranexamic acid, and aminocaproic acid involving children undergoing cardiac or scoliosis surgery. STUDY SELECTION AND DATA EXTRACTION: Twenty-three cardiac studies, totaling 1893 patients, met the inclusion criteria. None of the studies directly compared aprotinin to an alternative antifibrinolytic. Five scoliosis studies, totaling 207 patients, met the inclusion criteria. Data on blood loss and use of blood products in the first 24 postoperative hours were extracted. Only homogenously distributed outcomes were pooled. DATA SYNTHESIS: Tranexamic acid showed a homogeneously distributed reduction of blood loss by 11 mL/kg (95% confidence interval [CI] 9-13 mL/kg). Outcomes of blood loss reduction by aprotinin and aminocaproic acid were too heterogeneously distributed to be pooled, so the effect on blood loss could not be evaluated. Both aprotinin and tranexamic acid significantly reduced packed red cell transfusion (4 mL/kg, 95% CI 2-7 mL/kg and 7 mL/kg, 95% CI 5-10 mL/kg, respectively). Type of antifibrinolytic was not a determining factor that explained differences in outcome among trials in a meta-regression analysis. In the scoliosis studies, aprotinin and tranexamic acid significantly reduced blood loss compared with placebo (385 mL, 95% CI 727-42 mL and 682 mL, 95% CI 1149-214 mL, respectively). CONCLUSIONS: There is no evidence that suggests that, compared with aprotinin, alternative antifibrinolytics such as tranexamic acid were less effective in reducing blood loss in major pediatric surgery. PMID- 19188876 TI - A multi-institutional high-fidelity simulation "boot camp" orientation and training program for first year pediatric critical care fellows. AB - OBJECTIVE: Simulation training has been used to integrate didactic knowledge, technical skills, and crisis resource management for effective orientation and patient safety. We hypothesize multi-institutional simulation-based training for first year pediatric critical care (PCC) fellows is feasible and effective. DESIGN: Descriptive, educational intervention study. SETTING: The simulation facility at the host institution. INTERVENTIONS: A multicentered simulation-based orientation training "boot camp" for first year PCC fellows was held at a large simulation center. Immediate posttraining evaluation and 6-month follow-up surveys were distributed to participants. MEASUREMENTS AND MAIN RESULTS: A novel simulation-based orientation training for first year PCC fellows was facilitated by volunteer faculty from seven institutions. The two and a half day course was organized to cover common PCC crises. High-fidelity simulation was integrated into each session (airway management, vascular access, resuscitation, sepsis, trauma/traumatic brain injury, delivering bad news). Twenty-two first year PCC fellows from nine fellowship programs attended, and 13 faculty facilitated, for a total of 15.5 hours (369 person-hours) of training. This consisted of 2.75 hours for whole group didactic sessions (17.7%), 1.08 hours for a small group interactive session (7.0%), 4.67 hours for task training (30.1%), and 7 hours for training (45.2%) with high-fidelity simulation and crisis resource management. A "train to success" approach with repetitive practice of critical assessment and interventional skills yielded higher scores in training effectiveness in the end of-course evaluation. A follow-up survey revealed this training was highly effective in improving clinical performance and self-confidence. CONCLUSIONS: The first PCC orientation training integrated with simulation was effective and logistically feasible. The train to success concept with repetitive practice was highly valued by participants. Continuation and expansion of this novel multi institutional training is planned. PMID- 19188877 TI - Preventing corneal abrasions in critically ill children receiving neuromuscular blockade: a randomized, controlled trial. AB - OBJECTIVE: To determine the incidence of corneal abrasions in critically ill children requiring neuromuscular blockade (NMB) and to determine whether a moisture chamber over the eye is more effective in preventing corneal abrasions compared with standard therapy. DESIGN: Randomized controlled clinical trial. SETTING: Three pediatric intensive care units at two free-standing, university affiliated children's hospitals. PATIENTS: Consecutive intubated, mechanically ventilated patients receiving NMB <36 hours, 2 weeks to 18 years of age. INTERVENTION: After confirming the absence of a corneal abrasion, patients' eyes were randomized to either control (Q6H lubrication and eye closure) or treatment (Q6H lubrication, eyelid closure and moisture chamber). Eyes were examined daily for 3 days then every other day until NMB was discontinued, a corneal abrasion developed, or on study day 9. Data were analyzed on an intention-to-treat basis. MEASUREMENTS AND MAIN RESULTS: Two hundred thirty-seven patients were enrolled. Thirty patients exited the study before randomization (17 upon confirmation of a corneal abrasion on initial examination [7%; 95% confidence interval 4%-11%]; 12 upon discontinuation of NMB; 1 patient death). The remaining 207 patients were randomized and evaluated twice (median; interquartile range [IQR] 1-4 observations). Twenty-one patients developed a corneal abrasion over the course of the study (10%; 95%confidence interval 6%-15%). Median time from enrollment to abrasion was 2 days (IQR 1-3 days). The incidence of corneal abrasion was not different between the patient's control and treatment eyes; specifically, eight corneal abrasions developed in the control eye, five corneal abrasions in the treatment eye, and eight corneal abrasions in both eyes (McNemar's test; p = 0.58). CONCLUSIONS: The occurrence of corneal abrasions in critically ill children receiving NMB is not trivial. The additional use of a moisture chamber over the eye was no more effective than Q6H lubrication and eye closure alone in preventing corneal abrasions in this at-risk patient group. PMID- 19188878 TI - Simulation at the point of care: reduced-cost, in situ training via a mobile cart. AB - OBJECTIVE: The rapid growth of simulation in health care has challenged traditional paradigms of hospital-based education and training. Simulation addresses patient safety through deliberative practice of high-risk low-frequency events within a safe, structured environment. Despite its inherent appeal, widespread adoption of simulation is prohibited by high cost, limited space, interruptions to clinical duties, and the inability to replicate important nuances of clinical environments. We therefore sought to develop a reduced-cost low-space mobile cart to provide realistic simulation experiences to a range of providers within the clinical environment and to serve as a model for transportable, cost-effective, widespread simulation-based training of bona-fide workplace teams. DESIGN: Descriptive study. SETTING: A tertiary care pediatric teaching hospital. MEASUREMENTS AND MAIN RESULTS: A self-contained mobile simulation cart was constructed at a cost of $8054 (mannequin not included). The cart is compatible with any mannequin and contains all equipment needed to produce a high quality simulation experience equivalent to that of our on-site center--including didactics and debriefing with videotaped recordings complete with vital sign overlay. Over a 3-year period the cart delivered 57 courses to 425 participants from five pediatric departments. All individuals were trained among their native teams and within their own clinical environment. CONCLUSIONS: By bringing all pedagogical elements to the actual clinical environment, a mobile cart can provide simulation to hospital teams that might not otherwise benefit from the educational tool. By reducing the setup cost and the need for dedicated space, the mobile approach provides a mechanism to increase the number of institutions capable of harnessing the power of simulation-based education internationally. PMID- 19188879 TI - Lessons learned in the life of a pediatric intensivist. AB - The metamorphosis from a medical student to a professor is a complex process. It involves not only the acquisition of new knowledge but also the impact of countless human interactions throughout one's life. By necessity therefore, this evolution is a never ending process of enrichment of the mind and the soul. An important contributor to this process is learning from the perspectives developed by others based on their experiences. I describe seven valuable lessons learned during my life that may be helpful to the developing pediatric intensivists. PMID- 19188880 TI - Cost analysis of two approaches to parenteral nutrition in critically ill children. AB - OBJECTIVE: In pediatric intensive care, two approaches to parenteral nutrition are available: individualized admixtures or commercial standard solutions. Even though individualized admixtures can be tailored to sometimes highly intricate requirements, standard solutions are able to meet the demands of the majority of pediatric patients. To address the growing importance of costs in intensive care, we investigated whether relevant differences between individualized admixtures and standard solutions in terms of costs can be found. DESIGN: Retrospective analysis. SETTING: University-affiliated intensive care unit. PATIENTS: Fifty consecutively admitted pediatric patients requiring parenteral nutrition. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Comparison of actual costs of individualized admixtures and theoretical costs of standard solutions. Mean actual costs of individualized admixtures of Euro 81.78 (+/- 16.33) per patient and day were significantly higher than the theoretical costs of standard solutions of Euro 61.21 (+/- 6.55). Cost differences increased with patients' body weights. CONCLUSIONS: Parenteral nutrition with standard solutions offers the potential of a relevant cost reduction compared with individualized admixtures in critically ill children. PMID- 19188881 TI - Impact of a pediatric donation after cardiac death program. AB - OBJECTIVES: To determine the impact of a pediatric donation after cardiac death (DCD) program on organ donation. DESIGN: Retrospective case series. SETTING: A free-standing children's hospital. PATIENTS: All ventilated pediatric intensive care unit patients who died between September 1, 2005 and April 30, 2007. INTERVENTIONS: Institution of a DCD program. MEASUREMENTS AND MAIN RESULTS: Data collected included clinical features, medical eligibility and consent for organ donation, as well as outcome for donation among eligible patients. One hundred ten patients who died in the pediatric intensive care unit and were treated with mechanical ventilation immediately before death were identified. Thirty-one patients met exclusion criteria, 26 patients were not referred, and 53 patients were evaluated for potential DCD by the organ procurement agency. The majority of patients had anoxia or trauma as their primary diagnosis. Family members initiated discussions regarding donation in 17% of evaluations. Sixty-eight percent of patients were deemed medically ineligible by the organ procurement agency. Of the 17 medically eligible patients, consent for donation was given in 9 cases (53%). Of these, 7 patients (41%) successfully donated; accounting for 37% of organ donors during the study period. Two families gave consent for donation which did not occur; one child did not die within the required time period and one could not be matched with any recipients. CONCLUSIONS: Although a small percentage of dying patients are eligible for and will undergo DCD, such a program can markedly increase the number of organ donors at a children's hospital. PMID- 19188882 TI - [Cytokines as markers of osteolysis in the diagnostics of patients with bone metastases]. AB - In normal bone there are two essential processes of bone turnover, resorption and formation, which are disrupted by bone metastases. Two types of bone metastases are known, i.e. osteolytic lesions with dominant bone resorption and osteosclerotic tumors with enhanced osteoblastic bone formation. Numerous cytokines and growth factors regulate the activity of osteoclasts and/or osteoblasts in endo- or paracrine ways, playing crucial roles in the processes of bone turnover. Bone metastases are often the consequences of certain malignant tumors, such as breast, prostate, lung, and renal cancer. The diagnosis of bone metastasis is essential for a determination of the clinical stage of cancer and appropriate treatment. Tumor markers are useful in diagnosis, prognosis, staging, and, especially, monitoring treatment. Tumor markers are also useful in detecting bone metastases. There is growing evidence that various cytokines, especially M CSF, TGFbeta, TNFalpha, and IL-6 and IL-7, may be new tumor markers useful in the diagnosis of neoplastic disease. The processes of bone turnover in normal bone and metastatic tumors as well as the significance of the most important cytokines in the development of osteolytic metastases and the possibility of their use in the diagnosis of the most frequent cancers presenting bone metastases are described in this article. PMID- 19188883 TI - [Recognition of pathogens by innate immunity]. AB - The contributions of NK (natural killer) cells, DCs (dendritic cells), PRRs (pathogen recognition receptors), and PAMPs (pathogen-associated molecular patterns) recognized by PRRs as well as the participation of intracellular signaling pathways in innate immunity are shown. The possible involvement of a newly described group of immune cells, IKDCs (interferon-producing DCs), in innate immunity which show both NK and DC activity is presented. This review describes the mechanisms of innate immunity with the likely participation of autophagy initiated after endosomal TLR7 (Toll-like receptor 7) activation. Autophagy is able to remove intracellular bacteria or viruses by stimulating type I IFNs (interferons). A balance between PRR, especially TLR, activation and inactivation is crucial to avoid an excessive inflammatory response, as occurs in autoimmune and infectious diseases. Apart from SOCS (suppressor of cytokine signaling proteins) or Tollip (Toll-interacting protein), the possible contribution of microRNA in the regulation of innate immunity is described. The discovery of miRNA is one of the major scientific milestones in recent years and has revolutionized the way we look at many different modes of gene regulation. PMID- 19188884 TI - [Transforming growth factor beta (TGF-beta): its structure, function, and role in the pathogenesis of systemic lupus erythematosus]. AB - TGF-beta is a cytokine of great importance in many common diseases because it takes part in many physiological processes such as angiogenesis and stimulation of the synthesis and degradation of extracellular matrix proteins. It also regulates the entrance of cells to the apoptotic pathway and can stimulate the division of mesenchymal cells and inhibit hemopoietic, endothelial, and lymphatic cells. There are five genes which encode TGF-beta in vertebrates, of which only three are present in mammals. The best known member of the family of TGF-beta proteins is TGF-beta 1. TGF-beta is synthetized as a precursor protein which, after enzymatic modification, is present as a small or large complex. Three membrane receptors, serine/threonine kinase, are arranged for signal transduction with TGF-beta. Smad proteins are responsible for sending the signal into the cell nucleus; its influence on different transcriptive factors in the cell nucleus promotes the expressions of different genes. Disturbances in TGF-beta expression have been noted in many diseases. Current results clearly indicate an important role of this cytokine in autoimmunological disorders, especially in systemic lupus erythematosus. Studies on an animal model revealed that endogenic TGF-beta can control the progression of systemic lupus erythematosus. PMID- 19188885 TI - [Defensins and cathelicidins as natural peptide antibiotics]. AB - Defensins and cathelicidins belong to antimicrobial peptides (AMP), called also the natural antibiotics. They are found in Prokaryotes and Eukaryotes, also are synthesized in plants. These molecules were described in bacteria, invertebrates, vertebrates, also in mammals including humans. The amino-acids sequence is determined for about 880 antimicrobial peptides, and their classification is difficult and complex. These oligo- and polypeptides, are killing the microbes or inhibit their growth, are bactericidal to Gram-negative, Gram-positive bacteria, neutralizing toxins, demonstrated antiviral activity. AMP are multifunctional molecules, playing the first line of defense in human organism as effectors of unspecific immunity, with a broad spectrum of activity against pathogens, are characterized by controlling the physiological bacterial flora, often acting synergistically, are produced in different cells as defense against mechanisms of microbial resistance. They are chemotactic to neutrophiles and T cells, activate the dendritic cells. In human organism defensins are expressed in several cells and tissues, in neutrophiles, platelets, epithelial cells, in liver, skin, eye conjunctiva. Deficiency of defensins or cathelicidins cause pathological symptoms in atopic allergies or in Kostman disease. Several synthetic peptides are currently at the final phases of clinical trials regarding treatment of various bacterial infections. Defensins or cathelicidins could be also efficient as adjuvants or carriers for vaccines, in dairy industry as preservatives. This work presents some problems concerning the occurrence of AMP, especially defensins and cathelicidins of mammals, their classification, structure, various functions and mechanisms of bactericidal activity, and also their use in treatment of some infectious diseases. PMID- 19188886 TI - [T-cell receptor rearrangement excision circles (TRECs) as a marker of thymic function]. AB - The thymus is crucial in the reconstitution of the T-cell compartment following lymphodepletion and also in establishing a normal, diverse T-cell receptor (TCR) repertoire after immune response to antigens. TCRalphabeta diversity is generated through rearrangements of the TCR alpha and TCR beta chain genes. The TCR delta chain locus lies within the TCR alpha chain locus and its excision forms the first step in TCR alpha chain gene rearrangement. The intervening excised DNA is circularized by the formation of a "signal joint" forming a DNA episome, termed a signal-joint T-cell receptor excision circle (sjTREC). Approximately 70% of T cells emerging from the thymus contain one or two sjTRECs--depending on whether one or both TCRalpha loci genes are rearranged. As these long-lived naive T cells mature and proliferate, their sjTRECs are stable and do not divide. The thymus contributes naive T (CD45RA+) cells with TRECs to the peripheral immune system, but memory T cells (CD45RO+) contain few if any detectable TRECs. Quantification of thymic function is clinically relevant in settings with immunodeficiencies, after HSCT, during HIV therapy as well as in autoimmune patients. In humans there is no specific phenotypic marker for recent thymic emigrants, therefore the use of real-time quantitative PCR methods for absolute TREC quantification provides a good tool for estimating recent thymic function in different clinical situations. PMID- 19188887 TI - [Dendrimers in biomedical sciences and nanotechnology]. AB - Dendrimers are relatively new, hyper-branched polymers that have many interesting abilities. Dendrimers could be used, for example, as drug or gene carriers, contrast agents, sensors for different metal ions, and in developing innovation technology. These spherical polymers are also characterized by pharmacological activity against different bacterial and viral diseases. Dendrimers are currently being intensively investigated as anti-prion and anti-amyloid fibril agents. They can be used to build specific dendrimer films to be applied in modern technology. This review describes different uses of dendrimer particles in biomedical sciences and nanotechnology and shows advantages of their application. PMID- 19188888 TI - Genetic limits of sport performance: Quo vadis? PMID- 19188889 TI - Biomechanical and physiological comparison of barefoot and two shod conditions in experienced barefoot runners. AB - AIM: The first aim of this study was to assess how changes in the mechanical characteristics of the foot/shoe-ground interface affect spatio-temporal variables, ground pressure distribution, sagittal plane kinematics, and running economy in 8 experienced barefoot runners. The second aim was to assess if a special lightweight shoe (Vibram Fivefingers) was effective in mimic the experience of barefoot running. METHODS: By using an instrumented treadmill, barefoot running, running with the Fivefingers, and running with standard running shoe were compared, analyzing a large numbers of consecutive steps. Foot/shoe ground interface pressure distribution, lower limb kinematics, V.O(2) and heart rate data were simultaneously collected. RESULTS: Compared to the standard shod condition when running barefoot the athletes landed in more plantarflexion at the ankle. This caused reduced impact forces and changes in stride kinematics. In particular, significantly shorter stride length and contact times and higher stride frequency were observed (P<0.05). Compared to standard shod condition, V.O(2) and peak impact forces were significantly lower with Fivefingers (P<0.05) and much closer to barefoot running. Lower limb kinematics with Fivefingers was similar to barefoot running with a foot position which was significantly more plantarflexed than in control shoe (P<0.05). CONCLUSIONS: The data of this study support the assumption that changes in the foot-ground interface led to changes in running pattern in a group of experienced barefoot runners. The Fivefingers model seems to be effective in imitating the barefoot conditions while providing a small amount of protection. PMID- 19188890 TI - Polynomial modeling for the identification of lactate minimum velocity by different methods. AB - AIM: The lactate minimum (LM) protocol has been used to assess aerobic fitness and to predict exercise intensity associated with the maximal blood lactate steady state. The aim of this study was to compare different methods to identify the lactate minimum velocity (LMV) on cycling. METHODS: Fourteen male cyclists (26.8+/-4.5 years; 173.2+/-6.1 cm; 67.3+/-5.2 kg; 5,8+/-2.9 years of training) performed the LM test in a velodrome. The protocol consisted of an all out 2 km time trial to elevate blood lactate (bLAC), followed by 8 min of recovery and then 6 bouts of 2 km starting 5 kmxh(-1) below the individual mean velocity for the 6 km performance. The velocity was incremented by 1 kmxh(-1) at each bout with 25 microL of capillary blood being collected for bLAC measurements (YSI 2700 STAT). The LMV was identified visually (vLMV), and by applying a second grade polynomial function on 6 (pLMV(6)) and 3 (pLMV(3)) incremental bouts. Additionally, a method where the bLACx work velocity(-1) quotients (LMVQ) were plotted against the correspondent velocity during the incremental test, identified the LMV by considering 6 (LMVQ(6)) or 3 bouts (LMVQ(3)). RESULTS: ANOVA showed no differences between vLMV (33.1+/-2.5 kmxh(-1)), pLMV(6) (32.9+/ 2.5 kmxh(-1)), pLMV(3) (33.2+/-2.3 kmxh(-1)), LMVQ(6) (32.8+/-2.5 kmxh(-1)) and LMVQ(3) (33.4+/-2.3 kmxh(-1)), with high correlation among them. CONCLUSIONS: It was possible to identify the LMV by the methods proposed in the present study, even when the results of only 3 bouts of the test were modeled by polynomial function. Such an approach enables a more practical and economical test in addition to minimizing the discomfort due to several blood collections. PMID- 19188891 TI - Heart rate and blood lactate evaluation in bouldering elite athletes. AB - AIM: Bouldering contests consist of a series of short routes, called ''problems'', which shall be climbed without ropes. The purpose of this study was to determine blood lactate [Lac] concentration and heart rate (HR) adaptations both during an official bouldering competition and a simulated contest. METHODS: Eleven Italian elite climbers (6 males, 5 females) were tested from two bouldering national competitions (BNC). [Lac] assessments were done before and 2, 4, 6, 8 min after the contest. On a separate occasion, 9 different elite climbers (6 males, 3 females), matched for anthropometric features and ranking, were assessed in a simulated bouldering contest (SBC), under conditions and difficulties comparable to BNC. HR was measured through the whole performance; [Lac] was assessed before and 3 min after the achievement of each problem. RESULTS: The ascending time in both BNC and SBC was 391+/-85 s and 551+/-96 s (P<0.001), respectively. During SBC the mean HR peak of each problem was 93+/-8% of the subjects' age-predicted maximal HR (HRmax), and HR showed a full recovery after each problem, but in males only. Peak [Lac] after last problem was 6.6+/ 1.1 mmol/L and 5.6+/-1.1 mmol/L during BNC and SBC, respectively. A regression analysis showed that temporal changes in [Lac] depend on the attempts duration, with a critical cut-off time of 20 s. CONCLUSIONS: Bouldering contest requires an intense intermittent effort. The high HR may be due to the repeated isometric contractions, particularly of arm and forearm muscles. The net lactate production may depend on the attempts duration. PMID- 19188892 TI - The energy cost of ski mountaineering: effects of speed and ankle loading. AB - AIM: The purpose of this study is to determine the energy cost (EC) of ski mountaineering and its variation with speed and ankle loading. METHODS: Seven male skiers volunteered to participate in this study. Field tests (500 m, gradient 21%) were executed on packed snow at an altitude of about 1600 m. Measurements were carried out breath by breath by a portable gas analyzer. Energy cost of uphill skiing was calculated from the steady state VO(2). In the speed protocol each subject was asked to repeat the same route at three different speed levels. In the weight protocol, subjects were instructed to maintain the preferred speed for three trials on the same track while wearing different weight bands on their ankle. RESULTS: At the self-selected speed of 1.07+/-0.05 m s(-1) and without extra load beside the normal equipment, the mean value of EC on packed snow is 10.6+/-0.4 J kg(-1m)(-1). A percentage variation of the speed (%speed) produces a corresponding percentage variation of the energy cost %EC = 0.32 x %speed. The %EC as a function of the percentage of added load, %weight, with respect to the total weight of the subject, including ski, bindings, and boots is given by %EC = 1.71 x %weight. CONCLUSIONS: Data obtained in the present study constitute the first quantitative description of EC for ski mountaineering and result higher than for walking or snowshoeing. Effects due to ankle loading appear negligible for recreational skiers, while they should be taken into account in agonistic competition. PMID- 19188893 TI - Are physiological characteristics of Caribbean dance useful for health? AB - AIM: Although the current literature underlines the main role of physical inactivity in the development of chronic diseases and premature death, 65% of adults do not reach the minimum movement levels required to maintain and improve health. The aim of this study was to investigate whether the metabolic and cardiocirculatory characteristics of a single lesson of Caribbean dance fit with international recommendations to improve health through movement. METHODS: Energy expenditure, exercise intensity, mean heart rate and blood pressure response to a 90-minute lesson were analysed in 24 beginner and 24 experienced dancers (mean age 33.93+/-9.98 years). RESULTS: MANCOVA was used to analyse energy expenditure and exercise intensity of our sample, stratified for gender and experience. Body weight was inserted as a covariate. Experienced had a major total energy expenditure (372.75+/- 75.32 vs 297.33+/-87.54 kcal; P<0.001), minutes of exercise intensity >6 metabolic equivalents (METs) (8.04+/-10.65 vs 1.47+/-2.16; P<0.001), mean METs (3.81+/-0.32 vs 3.42+/-0.37; P<0.05) and mean heart rate (60.68+/-1.35 vs 53.24+/-3.91 %HRR; P<0.001) than beginner dancers. There were no statistical differences in blood pressure values. CONCLUSIONS: Caribbean dance fits with international guidelines to improve health and can aid the promotion and enhancement of health through its physiological characteristics, and may reduce drop-out due to a reduced motivation to move. PMID- 19188894 TI - The effects of a plyometric training program on the latency time of the quadriceps femoris and gastrocnemius short-latency responses. AB - AIM: The purpose of this study was to determine if a plyometric training program can affect the latency time of the quadriceps femoris and gastrocnemius short latency responses (SLRs) of the stretch reflex. METHODS: Sixteen healthy subjects (12 female and 4 male) were randomly assigned to either a control or a plyometric training group. Maximum vertical jump height (VJ) and SLRs of both quadriceps femoris and gastrocnemius were measured before and after a four week plyometric training program. RESULTS: Plyometric training significantly increased VJ (mean+/ SEM) by 2.38+/-0.45 cm (P<0.05) and non-significantly decreased the latency time of the quadriceps femoris SLR (mean+/-SEM) 0.363+/-0.404 ms (P>0.05) and gastrocnemius SLR (mean+/-SEM) 0.392+/-0.257 ms (P>0.05). VJ results support the effectiveness of plyometric training for increasing VJ height. CONCLUSIONS: The non-significant changes in the latency time of the quadriceps femoris and gastrocnemius SLRs seen in the training group suggest that performance improvements following a four-week plyometric training program are not mediated by changes in the latency time of the short-latency stretch reflex. PMID- 19188895 TI - Low bone mineral density and calcium intake in elite cyclists. AB - AIM: High physical activity is associated with larger bone mineral content (BMC) and density (BMD) in young males though competitive road cyclists have been reported to have similar or lower BMD than controls. METHODS: BMC and BMD were assessed in 73 highly trained (42 professional and 31 elite amateur) road cyclists (CYCLIST; age: 25.8+/-4.3 years; height: 179.7+/-6.3 cm; weight: 71.6+/ 6.3 kg; %Fat mass: 9.5+/-3 %; VO(2max): 68.5+/-5.7 mlxkg(-1)xmin(-1)) and in 30 healthy males used as reference (REF: 28.3+/-4.5 years; 176.6+/-6.2 cm; 74.5+/ 8.4 kg; 21.3+/-6.1%). RESULTS: Daily calcium intake estimated from a food questionnaire was higher in CYCLIST than in REF (942+/-374 vs 753+/-315 mgxd(-1); P=0.008). Compared to REF, CYCLIST had lower L1-L4 BMD (1.004+/-0.125 vs 1.240+/ 0.163 gxcm(-2); P<0.0001) and femoral neck BMD (0.986+/-0.132 vs 1.098+/-0.137; P<0.003). In CYCLIST, daily calcium intake is correlated with total BMC and BMD (r=0.27-0.26, P=0.02) and femoral BMD (r=0.35; P=0.002). Divided by tertiles (high, medium and low Ca), CYCLIST with high Ca (1320+/-382 mgxd(-1)) had lower lumbar BMD values (-6.68%; P=0.02) and tended to have lower femoral neck BMD ( 4.77%; P=0.09) and radius UD BMD (-5.9%; P=0.07) than REF. CONCLUSIONS: There was no difference between medium Ca and high Ca for any BMC or BMD parameters. Differences between low Ca and high Ca could be detected only for Total BMC ( 8.4%; P=0.01), Pelvis BMD (-7.4%; P=0.01) and femoral neck BMD (-9.9%; P=0.006). PMID- 19188896 TI - Weekly training hours are associated with molecular and cellular body composition levels in adolescent athletes. AB - AIM: We aimed to explore associations between hours per week of sports training and molecular and cellular body composition components in adolescent athletes. METHODS: A total of 33 female athletes (13.3+/-3.5 years; 47.8+/-12.6 kg; 154+/ 14.0 cm) and 90 male athletes (14.1+/-2.7 years; 60.6+/-17.8 kg; 167+/-16.2 cm) were measured. Based on the total of hours per week of training, athletes were divided into tertiles: <4.5 h/week; 4.5-8.9 h/week; 9 h/week. Dual-energy X-ray absorptiometry (DXA) was used to assess lean mass (lean), body fat (BF), percent body fat (%BF), bone mineral content (BMC) and density (BMD). Total body water (TBW), intracellular (ICW) and extracellular water (ECW) were assessed using bioelectrical impedance spectroscopy (BIS). Extracellular fluids (ECF), solids (ECS), body cell mass (BCM) and body fluid distribution (E/I) were calculated. Total hours per week of sports training (h/week), habitual physical activity (PA) and dietary were assessed by questionnaire. Statistics included analysis of covariance (ANCOVA) and simple regression analyses. RESULTS: Athletes training 9 h/week presented greater levels of TBW, lean, bone mass, BCM, and ECF and a lower %BF, independently of confounders. No significant differences in body composition estimates were found between athletes training <9 h/week. Hours per week of sports training were positively associated with fat free components, only in the group exercising 9 h/week CONCLUSIONS: In this group of Portuguese athletes from different sports we observed that training 9 h/week significantly improved body composition, especially fat free components, which may be important for a healthy growth and sports performance. PMID- 19188897 TI - Anterior cruciate ligament reconstruction with a quadrupled hamstrings tendon autograft does not restore tibial rotation to normative levels during landing from a jump and subsequent pivoting. AB - AIM: Recent research suggested that the anterior curciate ligament (ACL) reconstruction does not restore tibial rotation to normal levels when a bone patellar tendon bone (BPTB) graft is used during high demanding activities. Our goal was to determine if the usage of an alternative graft, as the quadrupled semitendinosus-gracilis (ST/G), restore tibial rotation to normal values in a population of athletically active individuals while performing a usual for their sport activity. METHODS: Eleven subjects, all reconstructed with an ST/G graft, were assessed in vivo, 9 months postoperatively, while they jumped off a 40 cm platform, landed on the ground and subsequently pivoted at 90 degrees. The evaluation period was identified from initial foot contact with the ground, included the pivoting of the ipsilateral leg, and was completed upon touchdown of the contralateral leg. By that time the patients had already returned to their sports activities. RESULTS: The maximum range of motion of the tibial rotation for the pivoting leg, during the evaluation period was found significantly (P=0.0001) larger in the reconstructed leg as compared to the intact contralateral, although both clinical and arthrometer assessments revealed restoration of anterior translation. CONCLUSIONS: It was concluded that ACL reconstruction with an ST/G graft does not restore tibial rotation to normal levels during this high demanding activity. It seems that new surgical techniques are needed to better replicate the actual anatomy and function of the natural ACL in order to address this problem. PMID- 19188898 TI - Injury profile of elite under-21 age female field hockey players. AB - AIM: The objective of this paper was to identify the rate, profile, and severity of injuries associated with participating on a provincial/state hockey (field) and compare these data, where possible, with available ice hockey data. METHODS: An injury was defined as ''any event, during team or team-related game, practice, and/or activity (on or off the playing surface), requiring any attention by the team's Therapist and/or Physician and subsequent game and/or practice time loss''. Seventy-five players, under the age of 21 years participated in the study over a 5-year duration. All injury data were collected post-injury. Data were collected on the player position, games versus practice conditions, injury time, injury type, injury etiology, anatomical region and plane injured, injury status, and duration required to return to full activity. RESULTS: A total of 2 828 athletes exposure's and 198 injuries were recorded. The combined injury rate was 70 injuries per 1 000 player game and practice exposures with significantly higher risk of injury resulting during the second half of a game or practice. Backs experienced the highest percentage and have a higher risk of injuries. The predominant injuries sustained included muscle strains, followed by tendonitis, while the highest number of injuries resulted from no contact. The lower back and ankle/foot were the most vulnerable to injury, followed by the knee. CONCLUSION: From this study it can be concluded that hockey (field) players can experience higher injury rates than ice hockey. Also, field hockey players are at greater risk of injury depending on the playing position and are more likely to be injured during the latter duration of a game and/or practice. In identifying injury trends related to hockey, injury prevention strategies should be developed as players use limited protective equipment. PMID- 19188899 TI - How do contralateral calcaneal quantitative ultrasound measurements in male professional football (soccer) players reflect the effects of high-impact physical activity on bone? AB - AIM: The aim of this study was to examine how the influence of football (soccer) on bone tissue was reflected by the acoustic parameters of bone as measured by calcaneal quantitative ultrasound (QUS) at both heels. We hypothesized that football players have less contralateral variability in QUS measurements than that in normally active individuals due to the bilateral impact loading characteristics of football. METHODS: Eighteen professional football players, aged 18-31 years, and 36 age-matched non-athletic controls participated in this study. Calcaneal QUS parameters [broadband ultrasound attenuation (BUA) and speed of sound (SOS)] were measured at both heels in all subjects using a gel coupled QUS device. RESULTS: Football players had significantly higher BUA and SOS mean values (30.6% and 45.5% higher mean BUA and 3.38% and 3.87% higher mean SOS values, for the right and the left heels, respectively) than those in non athletic controls (P=0.0001). Significant side differences were found in the mean values of BUA in the non-athletic subjects (P=0.001), but not in the footballers (P=0.538). The football players showed slightly higher BUA and SOS mean values on the left than those on the right side. While the number of football players who showed higher BUA values on either side was similarly distributed, significantly greater number of normally active males showed higher BUA values on the right side. Absolute and percentage side differences for BUA and the percentage of subjects who showed a bilateral BUA difference exceeding coefficient of variation %, expected from the repeated measurements of the same foot, in footballers were significantly less than those in the non-athletic subjects. CONCLUSIONS: The results of the present study show that high-impact loading of both calcanei in football is reflected by acoustic parameters of bone both in terms of overall enhanced bone properties and also in terms of the improvement of bone properties (as mainly reflected by BUA) on the non-dominant side as well. PMID- 19188900 TI - Supervised exercise versus non-supervised exercise for reducing weight in obese adults. AB - AIM: The prevalence of obesity is rising. Because obesity is positively associated with many health related risks and negatively associated with life expectancy this is a threat to public health. Physical exercise is a well known method to lose fat mass. Due to shame of their appearance, bad general condition and social isolation, starting and continuing physical exercise tends to be problematic for obese adults. A supervised training program could be useful to overcome such negative factors. In this study we hypothesized that offering a supervised exercise program for obese adults would lead to greater benefits in body fat and total body mass reduction than a non-specific oral advice to increase their physical activity. METHODS: Thirty-four participants were randomised to a supervised exercise program group (N.=17) and a control group (N.=17). Fifteen candidates in the intervention group and 12 in the control group appeared for baseline measurements and bought an all inclusive sports pass to a health club for Euro 10, per month. The control group just received the oral advice to increase their physical activity at their convenience. The supervised exercise group received biweekly exercise sessions of 2 hours with an estimated energy expenditure of 2 500 kJ per hour. Both groups received no dietary advice. RESULTS: After 4 months the overall decrease in body mass in the intervention group was 8.0 kg (SD 6.2) and the decrease in body fat was 6.2 kg (SD 4.5). The control group lost 2.8 kg overall (SD 4.2) and the decrease in body fat was 1.7 kg (SD 3.1). Correction for differences between groups in gender and age by multiple linear regression analysis showed significantly greater loss of total body mass (P = 0.001) and fat mass (P =0.002) in the intervention group compared with the control group. CONCLUSIONS: Stimulation of physical activity alone seems to result in a slight short term body mass and fat mass reduction in obese adults who are eager to lose weight. Supervised exercise under supervision of a qualified fitness instructor leads to a larger decrease. PMID- 19188901 TI - Multiple-choice reaction and visual perception in female and male elite athletes. AB - AIM: The aim of this study was to determine multiple-choice reaction and visual perception in female and male elite athletes. METHODS: A total of 49 athletes (26 males, 23 females) with a mean age of 21.84 +/- 1.94 in the range of 1826 years participated in this study. Participants were college athletes participating in basketball, soccer, handball, volleyball, gymnastics, fencing and swimming. Two tests were applied to investigate the multiple-choice reaction and visual perception of athletes: Vienna Determination Test and Visual Pursuit Test. RESULTS: The results indicated that male athletes had lower total amount of incorrect responses to a stimulus, compared with female athletes and that individual sports had a higher total amount of omitted responses given to a stimulus, compared with team sports. For ''on time'' response, there were significant differences for individual and team sports in interval 3. Depending on the sport branch, significant differences were found between male and female athletes in the amount of omitted responses given to a stimulus response in interval 3. CONCLUSIONS: The results of this study support the importance of reaction time related to aspects of the sports branch. PMID- 19188902 TI - An investigation into the physical determinants of change of direction speed. AB - AIM: Change of direction speed (CODS) is an important attribute for many sports and is believed to be influenced by a variety of physical factors. However, there is a lack of consensus as to which physical attributes relate to CODS. The aim of this study was to examine the relationship of several physical attributes to CODS. METHODS: Thirty-eight subjects (mean+/-SD: age, 21.5+/-3.8 years; height, 1.77+/-0.07 m; mass, 77.5+/-13.9 kg) undertook tests of speed, CODS, strength and power. Running speed was assessed via a 25 m sprint with split times taken at 5, 20 and 25 m. CODS was assessed by a 505-test, which involves measuring the time to complete a 5 m out and back course. The strength and power tests included unilateral isokinetic concentric and eccentric knee extensor and flexor strength at 60 degrees /s and bilateral leg press, countermovement and drop jumps. Pearson's product moment correlation and co-efficients of determination were used to explore relationships amongst all variables. Multiple regression was used to determine the combined effects of significantly correlated variables on CODS. RESULTS: Stepwise multiple regression revealed that running speed explained 58% of the variance in CODS (F(1,33)=45.796, P<0.001) with the addition of eccentric knee flexor strength raising the value to 67% (F(1,32)=8.781, P=0.006). CONCLUSIONS: The results suggest that for basic improvements in CODS, athletes should seek to maximise their sprinting ability and enhance their eccentric knee flexor strength to allow effective neuromuscular control of the contact phase of the CODS task. PMID- 19188903 TI - Acute incremental exercise to maximal performance does not cause alterations in serum oxidant levels of healthy young individuals. AB - AIM: This study was designed to analyze serum oxidative stress (OS) levels in healthy young individuals performing a routine maximal aerobic exercise and to evaluate the correlation between OS levels and physiological parameters. METHODS: Serum OS levels were studied by thermochemiluminescence (TCL) parameters at rest and following maximal aerobic exercise in 85 healthy young subjects. Levels were measured by a real time on line TCL assay (higher TCL-Ratio and TCL-H3 = lower OS level). RESULTS: Aerobic capacity had no effect on baseline OS levels. Post exercise OS levels correlated with maximal oxygen uptake (V.O(2max)) (P<0.005), delta V.O(2) (V.O(2max)- V.O(2)rest) (P<0.005), anaerobic threshold (VTH) (P<0.01), and total oxygen uptake (especially O(2) after VTH), (P<0.005). TCL Ratio was related to total running time (P<0.01), as well. Post-exercise OS levels for the whole study group did not vary from baseline values. However, individuals with higher fitness level (V.O(2max) >percentile 60) had significantly lower values of TCL-H3 (P=0.04) and tended to have lower TCL-Ratio, indicating they had elevated OS levels. In a multivariate analysis OS level was most affected by V.O(2) after VTH (anaerobic phase of the test) (P=0.003; adjusted odds ratio of 3.41, 95% confidence interval: 1.55-7.48). CONCLUSIONS: In conclusion, acute incremental exercise to maximal performance does not cause alterations in serum oxidant levels of healthy young individuals. In healthy individuals performing maximal aerobic exercise, OS levels correlate with maximal aerobic power. PMID- 19188904 TI - Effect of dextran sulfate sodium and acute exercise on mouse intestinal inflammation and lymphocyte cytochrome c levels. AB - AIM: Athletes may experience gastrointestinal disturbances during intense exercise. Using a mouse model, we determined the influence of acute exercise (AE) and dextran sulfate sodium (DSS), a chemical known to induce intestinal inflammation, on: 1) inflammatory changes within small and large intestine, 2) extent of cell death as measured by cytochrome c levels in intestinal lymphocytes (IL) and 3) the effects of bovine lactoferrin (bLf), a dietary protein with anti inflammatory properties, on these parameters. METHODS: DSS was given as 5% w/v in water for 4 days. AE consisted of 3 bouts of 90 min of exhaustive treadmill exercise, each separated by 24 h, with sacrifice before, immediately after, or 24 h after the final exercise bout. Mice were fed 2% bLf or control diet for 2 weeks before AE or DSS. Tissue inflammation was determined by histology and IL cytochrome c levels by Western blotting. RESULTS: AE increased plasma 8-iso PGF2a, a marker of oxidative stress, immediately after relative to before exercise (P<0.01). Cytochrome c levels were elevated following bLf (P<0.01) and DSS (P<0.05) treatment whereas AE had no significant effect. DSS, but not AE, produced histological changes suggestive of intestinal inflammation with no attenuation by bLf. CONCLUSIONS: Three bouts of AE were not associated with intestinal inflammation or IL death in this animal model. Gastrointestinal disturbances arising from intense exercise in humans may not be due to direct inflammatory damage although this remains to be determined clinically. PMID- 19188905 TI - Oxidative stress and antioxidant changes during a 24-hours mountain bike endurance exercise in master athletes. AB - AIM: This work monitored changes in oxidative stress and antioxidant defence during an endurance exercise in over 40 years old athletes. METHODS: Subjects were monitored during the 24-hours mountain bike Idro Lake (North of Italy) competition which took place in June 2008. The race lasted for 24 h, starting at 10.00 a.m., ending at 10.00 a.m. of the following day and was based upon riding for as many kilometers as possible in the 24-hours time schedule in a 5.5 km circuit trail. The study included 6 men bikers, aged 44.8 +/- 2 years, who raced on an individual basis. Blood samples were collected and the oxidative stress was measured performing the d-ROMs test which determined the reactive oxygen metabolites (ROMs), whereas the antioxidant defence status was assessed determining the biological antioxidant potential (BAP test). RESULTS: The ROMs levels significantly increased after 8 h from the beginning of the competition (122 %), at the end of the race (162%), 24 h (158%) and 48 h (144%) post-race. The biological antioxidant potential significantly increased at the end of the race (128%) and remained elevated 48 h later (114%). After 72 h post-race, ROMs and BAP levels differed significantly amongst subjects, thus showing an individual response to oxidative stress. CONCLUSIONS: In conclusion, exposure to intense and prolonged exercise induced a marked increase in dROMs levels in master athletes, only partially counterbalanced by antioxidants in blood plasma. The long-term effects of oxidative agents on the human body requires further studies, but it is likely that a diet potentially rich in antioxidants would help preventing oxidative damage of body cells and tissues and enhancing recovering from the endurance performance. PMID- 19188906 TI - Both tacrolimus and sirolimus decrease Th1/Th2 ratio, and increase regulatory T lymphocytes in the liver after ischemia/reperfusion. AB - The protective effects of immunosuppressants against ischemia/reperfusion (I/R) injury have been attributed to their non-specific anti-inflammatory effect. However, these effects may also depend on their effect on T lymphocytes, which are increasingly considered to be key players in I/R. Here, we studied the effects of tacrolimus and sirolimus on lymphocyte subpopulations in an I/R rat model. The animals were treated with tacrolimus, sirolimus or vehicle, before undergoing a 60-min ischemia event of the right hepatic lobe, followed by excision of the remaining liver. After 2 h, I/R rats showed increased mortality, plasma lactate dehydrogenase (LDH) levels, hepatocyte apoptosis, liver histological injury and parenchymal infiltration by neutrophils, macrophages, NK cells and T lymphocytes. Most of the changes were antagonized by both immunosuppressants. Tacrolimus augmented the proportion of cycling cells in I/R rats, whereas sirolimus showed the opposite effect. The increased Th1/Th2 ratio found in I/R livers after 2 h was reverted by immunosuppressants, which also amplified the proportion of CD4(+)CD25(+)Foxp3(+) regulatory T lymphocytes at 24 h. The protective effects of both tacrolimus and sirolimus correlated well with a decreased ratio of proinflammatory to anti-inflammatory T lymphocytes, and with an increase in the Treg proportion. This suggests a new mechanism to explain the known beneficial effect shown by immunosuppressants early after I/R. PMID- 19188907 TI - Human peritoneal fibroblasts are a potent source of neutrophil-targeting cytokines: a key role of IL-1beta stimulation. AB - Polymorphonuclear leukocyte (PMN) infiltration is a cardinal feature of peritonitis. CXC chemokine ligands 1 and 8 (CXCL1 and CXCL8), and the cytokine granulocyte colony-stimulating factor (G-CSF) are the key mediators of PMN accumulation. Increasing evidence points to an important role of human peritoneal fibroblasts (HPFB) in the response of the peritoneum to infection. We have examined the synthesis of PMN-targeting cytokines by HPFB exposed to intraperitoneal milieu as represented by peritoneal dialysate effluent (PDE) from patients undergoing peritoneal dialysis. PDE obtained during peritonitis, but not during infection-free periods, significantly increased production of CXCL1, CXCL8, and G-CSF by HPFB. The effect was largely blocked by antibodies to interleukin-1beta (IL-1beta), whereas neutralization of tumor necrosis factor alpha (TNFalpha) had no major effect. Similar pattern of inhibition was observed when HPFB were exposed to conditioned media from endotoxin-stimulated peritoneal macrophages. Significance of IL-1beta stimulation was further shown in experiments with recombinant cytokines. Compared with TNFalpha, exposure of HPFB to recombinant IL-1beta resulted in a much higher release of PMN-targeting cytokines. The assessment of mRNA degradation revealed that the IL-1beta-induced transcripts of CXCL1, CXCL8, and G-CSF were more stable compared with those induced by TNFalpha. These data indicate that HPFB can be a significant source of PMN-targeting cytokines when stimulated with IL-1beta in the inflamed peritoneum. PMID- 19188908 TI - Rapid diagnosis of human prion disease using streptomycin with tonsil and brain tissues. AB - The use of streptomycin in the pathological prion protein (PrP(sc)) detection procedures represents a new and attractive way for diagnostic purpose. With this agent, western blot readily detected PrP(sc) in 263K scrapie hamster and C57Bl/6 wild-type mice challenged with C506M3 scrapie strain. Our aim was to evaluate this new diagnosis procedure in the field of human transmissible spongiform encephalopathies (TSEs). First, we had confirmed the ability of streptomycin to precipitate PrP(res) from human brain of Creutzfeldt-Jakob disease (CJD) patient. Second, we compared the detection of PrP(res) with streptomycin against three other protocols using other precipitations. Then we assessed PrP(res) detection with streptomycin in 98 brain tissue samples from various aetiologies of human TSEs and 52 brain samples from other dementia. Finally, we applied this protocol for tonsils examination of five patients suspected of variant CJD (v-CJD). Sensitivity and specificity obtained with the streptomycin protocol were both 100% on brain tissue. For tonsil tissues, PrP(res) was clearly identified in the two post-mortem confirmed v-CJD cases, whereas no characteristic three-band pattern was seen in the three confirmed non-v-CJD samples. In this study, streptomycin demonstrated its efficiency to detect PrP(res) both in the central nervous system and in the lymphoid tissue without practical difficulty and with rapid preparation. Because of its ability to act as a good agent for PrP(sc) examination in different tissues, recovery of PrP(sc) in biological fluids using streptomycin should open further perspectives of applications in CJD diagnostics. Streptomycin effects in vivo might thus also be questioned. PMID- 19188909 TI - Anaphylaxis to a self-peptide in the absence of mast cells or histamine. AB - Induction of T helper 1 (Th1) to Th2 deviation through administration of self- or altered self-peptides holds promise for treatment of autoimmunity. However, administration of self-peptides in models of autoimmunity can result in anaphylactic reactions. Although both IgE and IgG1 antibodies might be involved in the development of anaphylaxis to myelin peptides in experimental autoimmune encephalomyelitis in mice, the effector cells and molecules involved are not fully understood. Here we show that systemic anaphylaxis to the self-antigen myelin oligodendrocyte glycoprotein (MOG) 35-55 can occur in mice lacking mast cells (Kit(W)/Kit(W-v) mice) or histamine (histidine decarboxylase-deficient mice), but is prevented in mice lacking IL-4. Treatment of mice with CV6209, a platelet-activating factor antagonist, slightly reduced the incidence of anaphylaxis to self-MOG35-55 in this model, but more effectively protected mice against anaphylaxis to this peptide when self-MOG35-55 was administered in a different immunization protocol that omitted the use of Bordetella pertussis toxin as an adjuvant at the time of immunization. Thus, anaphylactic reactions to self-MOG can occur in the absence of mast cells or histamine, key elements of the classical IgE-, mast cell-, and histamine-dependent pathway of anaphylaxis. PMID- 19188910 TI - Interstitial fibrosis and microvascular disease of the heart in uremia: amelioration by a calcimimetic. AB - In patients with chronic renal failure, the heart undergoes remodeling, characterized by hypertrophy, fibrosis, and capillary/myocyte mismatch. In this study, we observed the effects of the calcimimetic agent R-568 on microvascular disease and interstitial fibrosis of the heart. Three-month-old male Sprague Dawley rats were randomized to subtotal nephrectomy (SNX) or sham operation and subsequently received vehicle or R-568 under two experimental protocols, one for 1 month and the other for 3 months. Echocardiography, capillary length density, volume density of interstitial tissue, and immunohistochemistry and western blots (calcium-sensing receptor, collagen I and III, transforming growth factor (TGF) beta, mitogen-activated protein kinases, and nitrotyrosine) were assessed. After SNX, weight and wall thickness of the left and the right ventricle were elevated. The ratio of heart to body weight and interventricular septum thickness were not changed by R-568 treatment. The left ventricle fractional shortening (by echocardiography) was lower in SNX; this was ameliorated by R-568. Reduced capillary length density and increased interstitial fibrosis in SNX were improved by R-568, which also reduced the expression of TGF-beta, and collagen I and III. The calcimimetic increased the activation of ERK-1/2, normalized p38 and JNK signaling, and prevented oxidative stress. We conclude that lowering parathyroid hormone with a calcimimetic significantly improves cardiac histology and function but not the left ventricular mass in SNX. PMID- 19188911 TI - Adenylate cyclase-associated protein 1 overexpressed in pancreatic cancers is involved in cancer cell motility. AB - Pancreatic cancer has the worst prognosis among cancers due to the difficulty of early diagnosis and its aggressive behavior. To characterize the aggressiveness of pancreatic cancers on gene expression, pancreatic cancer xenografts transplanted into severe combined immunodeficient mice served as a panel for gene expression profiling. As a result of profiling, the adenylate cyclase-associated protein 1 (CAP1) gene was shown to be overexpressed in all of the xenografts. The expression of CAP1 protein in all 73 cases of pancreatic cancer was recognized by immunohistochemical analyses. The ratio of CAP1-positive tumor cells in clinical specimens was correlated with the presence of lymph node metastasis and neural invasion, and also with the poor prognosis of patients. Immunocytochemical analyses in pancreatic cancer cells demonstrated that CAP1 colocalized to the leading edge of lamellipodia with actin. Knockdown of CAP1 by RNA interference resulted in the reduction of lamellipodium formation, motility, and invasion of pancreatic cancer cells. This is the first report demonstrating the overexpression of CAP1 in pancreatic cancers and suggesting the involvement of CAP1 in the aggressive behavior of pancreatic cancer cells. PMID- 19188913 TI - Blockade of the c-Jun amino terminal kinase prevents crescent formation and halts established anti-GBM glomerulonephritis in the rat. AB - Macrophages induce acute renal injury in anti-glomerular basement membrane (GBM) glomerulonephritis. This operates, in part, via activation of the c-Jun amino terminal kinase (JNK) signaling pathway. However, it is unknown whether inhibition of JNK signaling is effective once the proinflammatory response is established in the injured kidney. This study examined whether blockade of JNK signaling could halt disease progression, including crescent formation, in a model of severe crescentic anti-GBM glomerulonephritis. WKY rats were immunized with sheep IgG and then injected with sheep anti-GBM serum (day 0). Animals were treated with the JNK inhibitor, CC-401, vehicle alone, or no treatment from day 7 until being killed on day 24 of disease. Untreated animals at day 7 showed significant proteinuria, focal glomerular lesions, marked glomerular macrophage and T-cell accumulation, and upregulation of proinflammatory mediators (TNF alpha, iNOS, MMP-12). Untreated and vehicle-treated groups displayed severe glomerulonephritis at day 24 with renal impairment and worsening proteinuria. These animals had severe glomerular lesions, with 60% of glomeruli exhibiting fibrocellular crescents, in association with increased macrophage and T-cell accumulation (including macrophage giant cells) and a further increase in mRNA levels of TNF-alpha, iNOS, MMP-12, and TGF-beta1. In contrast, CC-401 treatment prevented renal impairment, suppressed proteinuria, and prevented severe glomerular and tubulointerstitial lesions, including crescent formation and granulomatous-like lesions. These protective effects were independent of glomerular macrophage and T-cell accumulation, and of the humoral immune response. CC-401 treatment inhibited expression of both pro- and antiinflammatory molecules (interleukin-10 and heme oxygenase-1). In addition, IL-1 induced MMP-12 and IL-10 production by cultured macrophages was found to be JNK dependent. In conclusion, blockade of JNK signaling provides substantial protection against the progression of crescentic anti-GBM glomerulonephritis, which may be, in part, due to inhibition of the macrophage proinflammatory response. PMID- 19188914 TI - Darwin's legacy. PMID- 19188912 TI - Sepsis induces extensive autophagic vacuolization in hepatocytes: a clinical and laboratory-based study. AB - Autophagy is the regulated process cells use to recycle nonessential, redundant, or inefficient components and is an adaptive response during times of stress. In addition to its function in enabling the cell to gain vital nutrients in times of stress, autophagy can also be involved in elimination of intracellular microorganisms, tumor suppression, and antigen presentation. Because of difficulty in diagnosing autophagy, few clinical studies have been performed. This study examined whether autophagy occurs in hepatocytes during sepsis. Electron microscopy (EM) was performed on liver samples obtained from both an observational clinical cohort of six septic patients and four control patients as well as liver specimens from mice with surgical sepsis (by cecal ligation and puncture) or sham operation. EM demonstrated increased autophagic vacuoles in septic vs nonseptic patients. Randomly selected fields (3000 microm(2)) from control and septic patients contained 1.2+/-1.5 vs 5.3+/-3.3 (mean+/-s.d.) complex lysosomal/autophagolysosomal structures per image respectively (P<0.001). In rare instances, hepatocytes with autophagic vacuoles appeared to be unequivocally committed to death. Membrane alterations (membrane vacuoles, invagination into adjacent organelles, and myelin figure-like changes) occur in a subpopulation of mitochondria in sepsis, but other hepatocyte organelles showed no consistent ultrastructural injury. Findings in murine sepsis paralleled those of patients, with 7.2+/-1.9 vs 38.7+/-3.9 lysosomal/autophagolysosomal structures in sham and septic mice, respectively (P=0.002). Quantitative RT-PCR demonstrated that sepsis induced the upregulation of select apoptosis and cytokine gene expression with minimal changes in the core autophagy genes in liver. In conclusion, hepatocyte autophagic vacuolization increases during sepsis and is associated with mitochondrial injury. However, it is not possible to determine whether the increase in autophagic vacuolization is an adaptive response or a harbinger of cell death. PMID- 19188915 TI - Cytokinesis: mind the GAP. PMID- 19188916 TI - Alzheimer's dementia by circulation disorders: when trees hide the forest. PMID- 19188917 TI - Linear polyubiquitylation: the missing link in NF-kappaB signalling. PMID- 19188918 TI - Casting back to stem cells. PMID- 19188921 TI - Genome-wide association analyses suggested a novel mechanism for smoking behavior regulated by IL15. AB - Cigarette smoking is the leading preventable cause of death in the United States. Although smoking behavior has a significant genetic determination, the specific genes and associated mechanisms underlying the smoking behavior are largely unknown. Here, we carried out a genome-wide association study on smoking behavior in 840 Caucasians, including 417 males and 423 females, in which we examined approximately 380,000 single nucleotide polymorphisms (SNPs). We found that a cluster of nine SNPs upstream from the IL15 gene were associated with smoking status in males, with the most significant SNP, rs4956302, achieving a P-value (8.80 x 10(-8)) of genome-wide significance. Another SNP, rs17354547 that is highly conserved across multiple species achieved a P-value of 5.65 x 10(-5). These two SNPs, together with two additional SNPs (rs1402812 and rs4956396) were selected from the above nine SNPs for replication in an African-American sample containing 1251 subjects, including 412 males and 839 females. The SNP rs17354547 was replicated successfully in the male subgroup of the replication sample; it was associated with smoking quantity (SQ), the Heaviness of Smoking Index (HSI) and the Fagerstrom Test for Nicotine Dependence (FTND), with P-values of 0.031, 0.0046 and 0.019, respectively. In addition, a haplotype formed by rs17354547, rs1402812 and rs4956396 was also associated with SQ, HSI and FTND, achieving P values of 0.039, 0.0093 and 0.0093, respectively. To further confirm our findings, we carried out an in silico replication study of the nine SNPs in a Framingham Heart Study sample containing 7623 Caucasians from 1731 families, among which, 3491 subjects were males and 4132 were females. Again, the male specific association with smoking status was observed, for which seven of the nine SNPs achieved significant P-values (P<0.05) and two achieved marginally significant P-values (P<0.10) in males. Several of the nine SNPs, including the highly conserved one across species, rs17354547, are located at potential transcription factor binding sites, suggesting transcription regulation as a possible function for these SNPs. Through this function, the SNPs may modulate the gene expression of IL15, a key cytokine regulating immune function. As the immune system has long been recognized to influence drug addiction behavior, our association findings suggest a novel mechanism for smoking addiction involving immune modulation through the IL15 pathway. PMID- 19188922 TI - Long non-coding RNAs: insights into functions. AB - In mammals and other eukaryotes most of the genome is transcribed in a developmentally regulated manner to produce large numbers of long non-coding RNAs (ncRNAs). Here we review the rapidly advancing field of long ncRNAs, describing their conservation, their organization in the genome and their roles in gene regulation. We also consider the medical implications, and the emerging recognition that any transcript, regardless of coding potential, can have an intrinsic function as an RNA. PMID- 19188924 TI - Obesity does not accelerate the decline in glomerular filtration rate associated with advancing age. AB - Obesity has been suggested as a risk factor for chronic kidney disease. However, it has also been suggested that the association between obesity and impaired glomerular filtration rate (GFR) arises from the invalid use of body surface area (BSA) for scaling. This study assesses the effect of obesity on GFR by comparing the age-dependent decline in obese (body mass index (BMI) >30 kg/m(2); n=149) and non-obese patients (n=589), aged >30 years, referred for measurement of GFR (Cr 51-EDTA and three blood samples). GFR was scaled to a BSA of 1.73 m(2) (GFR/BSA) and extracellular fluid volume of 13 l (GFR/ECV), both corrected for the one compartment assumption. When non-obese patients were categorized into 10-year age brackets (from 31 to >70), GFR/BSA and GFR/ECV declined from 92 ml per min per 1.73 m(2) and 95 ml per min per 13 l, respectively, at 31-40 years to 58 and 59 at >70. The declines in obese patients were similar with corresponding values of 88 ml per min per 1.73 m(2) and 97 ml per min per 13 l at 31-40 and 57 and 59 at >70 years. Linear regression analysis of non-categorized data from age 40 years showed rates of decline slightly slower in the obese (0.82 vs 0.95 ml per min per 1.73 m(2) per year and 0.87 vs 1.02 ml per min per 13 l per year). No effect of obesity on renal function was shown. Scaling to BSA did not distort the results. PMID- 19188923 TI - The consequences of asynapsis for mammalian meiosis. AB - During mammalian meiosis, synapsis of paternal and maternal chromosomes and the generation of DNA breaks are needed to allow reshuffling of parental genes. In mammals errors in synapsis are associated with a male-biased meiotic impairment, which has been attributed to a response to persisting DNA double-stranded breaks in the asynapsed chromosome segments. Recently it was discovered that the chromatin of asynapsed chromosome segments is transcriptionally silenced, providing new insights into the connection between asynapsis and meiotic impairment. PMID- 19188925 TI - Ghrelin restores 'lean-type' hunger and energy expenditure profiles in morbidly obese subjects but has no effect on postgastrectomy subjects. AB - OBJECTIVE: To examine the effects of ghrelin on appetite and energy expenditure in lean, obese and postgastrectomy subjects. DESIGN: A randomized, double-blind, placebo-controlled study. PATIENTS: Nine lean subjects (mean body mass index (BMI) 23.5+/-3 kg/m(2)) and nine morbidly obese subjects (mean BMI 51.4+/-10 kg/m(2)) and eight postgastrectomy subjects (mean BMI 22.4+/-1.0 kg/m(2)). INTERVENTIONS: Subjects were infused with either intravenous ghrelin (5 pmol kg( 1) min(-1)) or saline over 270 min. They were given a fixed energy breakfast followed by a free buffet lunch towards the end of the infusion. MAIN OUTCOME MEASURES: Visual analogue scales were used to record hunger and energy expenditure was measured by indirect calorimetry. RESULTS: Ghrelin increased energy intake at the buffet lunch in lean subjects (a 41% increase, P<0.01) and obese subjects (35% increase, P=0.04) but not in postgastrectomy subjects. Lean subjects showed a characteristic preprandial rise and postprandial fall in hunger scores, which was exaggerated by ghrelin infusion. Obese subjects showed little variation in hunger scores, but a 'lean-type' pattern was restored when given exogenous ghrelin. Ghrelin had no effect on resting metabolic rate but did increase respiratory quotient (RQ) in obese subjects. Ghrelin also increased RQ variability over time in all three groups (ANOVA, P<0.001). CONCLUSIONS: Hunger scores are abnormal in the obese, perhaps because of impaired ghrelin secretion. The effect of ghrelin in restoring normal hunger profiles in the obese suggests causality, confirming an important role in eating behaviour. Ghrelin also increases RQ in obese humans and increased RQ variability in all groups. This suggests that ghrelin regulates substrate utilization and may promote metabolic flexibility. PMID- 19188926 TI - Subtyping obesity with microarrays: implications for the diagnosis and treatment of obesity. AB - OBJECTIVE: Obese patients respond differently to weight loss interventions. No efficient diagnostic tool exists to separate obese patients into subtypes as a means to improve prediction of response to interventions. We aimed to separate obese subjects into distinct subgroups using microarray technology to identify gene expression-based subgroups to predict weight loss. DESIGN: A total of 72 obese men and women without family history of diabetes were enrolled in the study; 52 were treated with ephedra and caffeine (E+C) and 20 with placebo for 8 weeks. Adipose and skeletal muscle tissue biopsies were performed at baseline. RNA sample pairs were labeled and hybridized to oligonucleotide microarrays. Quantile normalization and gene shaving were performed, and a clustering algorithm was then applied to cluster subjects based on their gene expression profile. Clusters were visualized using heat maps and related to weight changes. RESULTS: Cluster analysis of gene expression data revealed two distinct subgroups of obesity and predicted weight loss in response to the treatment with E+C. One cluster ('red') decreased to 96.87+/-2.35% body weight, and the second cluster ('green') decreased to 95.59+/-2.75% body weight (P<0.05). 'Red' cluster had less visceral adipose tissue mass (2.77+/-1.08 vs 3.43+/-1.49 kg; P<0.05) and decreased size of the very large fat cells (1.45+/-0.61 vs 2.16+/-1.74 microl; P<0.05) compared to 'green' cluster. Gene expression for both skeletal muscle and adipose tissue was also different between clusters. CONCLUSIONS: Our study provides the first evidence that the combined approach of gene expression profiling and cluster analysis can identify discrete subtypes of obesity, these subtypes have different physiological characteristics and respond differently to an adrenergic weight loss therapy. This brings us that into an era of personalized treatment in the obesity clinic. PMID- 19188927 TI - A proposed clinical staging system for obesity. AB - Current classifications of obesity based on body mass index, waist circumference and other anthropometric measures, although useful for population studies, have important limitations when applied to individuals in clinical practice. Thus, these measures do not provide information on presence or extent of comorbidities or functional limitations that would guide decision making in individuals. In this paper we review historical and current classification systems for obesity and propose a new simple clinical and functional staging system that allows clinicians to describe the morbidity and functional limitations associated with excess weight. It is anticipated that this system, when used together with the present anthropometric classification, will provide a simple framework to aid decision making in clinical practice. PMID- 19188928 TI - Intra-rater agreement of the anorectal exam and classification of injury severity in children with spinal cord injury. AB - STUDY DESIGN: Intra-rater reliability study, cross-sectional design. OBJECTIVES: To report on the intra-rater agreement of the anorectal examinations and classification of injury severity in children with spinal cord injury (SCI). SETTING: Two, non-profit children's hospitals specializing in pediatric SCI. METHODS: 180 subjects had at least two trials of the anorectal examinations as defined by the International Standards for Neurological Classification of Spinal Cord Injury. Intraclass correlation coefficients (ICC) and 95% confidence intervals (CI) were used to evaluate the agreement. ICC>0.90=high agreement; ICC between 0.75-0.89=moderate agreement; ICC<0.75=poor agreement. RESULTS: When evaluated for the entire sample, agreement was moderate-high for anal sensation and contraction and injury classification. When evaluated as a function of age at examination and type of injury, agreement for anal sensation was poor for subjects with tetraplegia in the 12-15-year age group (ICC=0.56) and 16-21-year age group (ICC=0.70) and for subjects with paraplegia in the 6-11-year age group (ICC=0.69). Agreement for anal contraction was moderate for subjects with tetraplegia in the 16-21-year age group (ICC=0.81) and subjects with paraplegia in the 12-15-year age group (ICC=0.78) and poor for subjects with paraplegia in the 6-11-year age group (ICC=0.67). Agreement for injury classification was poor for subjects with tetraplegia in the 12-15-year group (ICC=0.56) and 16-21-year group (ICC=0.74) and paraplegia in the 6-11-year group (ICC=0.11) and 12-15-year group (ICC=0.63). Anorectal responses had high agreement in subjects with tetraplegia in the 6-11-year group and moderate to high agreement in subjects with paraplegia in the 16-21-year group. CONCLUSION: The data do not fully support the use of anorectal examination in children. Further work is warranted to establish the validity of anorectal examination. PMID- 19188929 TI - Identification of genes that confer tumor cell resistance to the aurora B kinase inhibitor, AZD1152. AB - AZD1152 is a highly selective Aurora B kinase inhibitor currently undergoing Phase I and II clinical evaluation in patients with acute myelogenous leukemia and advanced solid malignancies. We have established two AZD1152-resistant cell lines from SW620 colon and MiaPaCa pancreatic carcinoma lines, which are >100 fold resistant to the active metabolite of AZD1152, AZD1152 HQPA and interestingly, cross-resistant to the pan-Aurora kinase inhibitor, VX-680/MK0457. Using whole-genome microarray analysis and comparative genomic hybridization, we were able to identify MDR1 and BCRP as the causative genes that underlie AZD1152 HQPA-resistance in these models. Furthermore, the upregulation of either of these genes is sufficient to render in vivo tumor growth insensitive to AZD1152. Finally, the upregulation of MDR1 or BCRP is predictive of tumor cell sensitivity to this agent, both in vitro and in vivo. The data provide a genetic basis for resistance to Aurora kinase inhibitors, which could be utilized to predict clinical response to therapy. PMID- 19188930 TI - Epigenetics: heterochromatin meets RNAi. AB - The term epigenetics refers to heritable changes not encoded by DNA. The organization of DNA into chromatin fibers affects gene expression in a heritable manner and is therefore one mechanism of epigenetic inheritance. Large parts of eukaryotic genomes consist of constitutively highly condensed heterochromatin, important for maintaining genome integrity but also for silencing of genes within. Small RNA, together with factors typically associated with RNA interference (RNAi) targets homologous DNA sequences and recruits factors that modify the chromatin, commonly resulting in formation of heterochromatin and silencing of target genes. The scope of this review is to provide an overview of the roles of small RNA and the RNAi components, Dicer, Argonaute and RNA dependent polymerases in epigenetic inheritance via heterochromatin formation, exemplified with pathways from unicellular eukaryotes, plants and animals. PMID- 19188931 TI - A retrograde apoptotic signal originating in NGF-deprived distal axons of rat sympathetic neurons in compartmented cultures. AB - Previous investigations of retrograde survival signaling by nerve growth factor (NGF) and other neurotrophins have supported diverse mechanisms, but all proposed mechanisms have in common the generation of survival signals retrogradely transmitted to the neuronal cell bodies. We report the finding of a retrograde apoptotic signal in axons that is suppressed by local NGF signaling. NGF withdrawal from distal axons alone was sufficient to activate the pro-apoptotic transcription factor, c-jun, in the cell bodies. Providing NGF directly to cell bodies, thereby restoring a source of NGF-induced survival signals, could not prevent c-jun activation caused by NGF withdrawal from the distal axons. This is evidence that c-jun is not activated due to loss of survival signals at the cell bodies. Moreover, blocking axonal transport with colchicine inhibited c-jun activation caused by NGF deprivation suggesting that a retrogradely transported pro-apoptotic signal, rather than loss of a retrogradely transported survival signal, caused c-jun activation. Additional experiments showed that activation of c-jun, pro-caspase-3 cleavage, and apoptosis were blocked by the protein kinase C inhibitors, rottlerin and chelerythrine, only when applied to distal axons suggesting that they block the axon-specific pro-apoptotic signal. The rottlerin sensitive mechanism was found to regulate glycogen synthase kinase 3 (GSK3) activity. The effect of siRNA knockdown, and pharmacological inhibition of GSK3 suggests that GSK3 is required for apoptosis caused by NGF deprivation and may function as a retrograde carrier of the axon apoptotic signal. The existence of a retrograde death signaling system in axons that is suppressed by neurotrophins has broad implications for neurodevelopment and for discovering treatments for neurodegenerative diseases and neurotrauma. PMID- 19188932 TI - Induction of CD4+CD25+Foxp3+ regulatory T cell response by glatiramer acetate in type 1 diabetes. AB - Glatiramer acetate (GA) is an immunomodulatory peptide drug used to treat multiple sclerosis. Its treatment effect has been expanded to other autoimmune conditions such as uveoretinitis, inflammatory bowel disease, graft rejection and hepatic fibrosis. Here, we report that GA was effective in altering the clinical course of diabetes in cyclophosphamide (CY)-potentiated non-obese diabetic (CY NOD) mice. Treatment with GA significantly reduced the diabetic rate in the mice and ameliorated insulitis, which coincided with increased CD4+CD25+Foxp3+ T cell response in treated mice. GA treatment led to increased expression of transcription factor Foxp3 and elevated production of interleukin-4 (IL-4) both in vivo and in vitro. It was evident that the effect of GA on up-regulation of Foxp3 was mediated partially through IL-4. IL-4 was found to maintain Foxp3 expression and regulatory function of CD4+CD25+ regulatory T cells (Tregs). This study provides new evidence that GA has treatment potential for type 1 diabetes through the induction of Tregs and that increased IL-4 production is partially responsible for the enhanced Treg's function in GA treatment. PMID- 19188933 TI - [Healthy people in healthy communities]. PMID- 19188934 TI - [Volatile organic compounds in ready-to-eat fruits and vegetable products]. AB - An increased consumer demand for bagged prepared fruits and vegetables has recently occurred, these being ready-to-eat products. The different phases in the preparation of these products include cleaning, peeling, cutting, washing, drying and packaging. The quality, safety and shelf-life of ready-to-eat products is highly influenced by the washing process which is generally performed by soaking the vegetables in cold water containing disinfectants (usually sodium hypochlorite). We therefore evaluated the presence of halogenated volatile organic compounds (VOC) in 70 samples of ready-to-eat products produced by 15 different establishments. Results showed that 54% of the products were contaminated by at least one halogenated VOC. Trialomethane was the most frequently detected contaminant and 50% of samples were found to contain chloroform. Contamination by other halogenated VOCs was less frequent. Also, there was variation in concentration values of contaminants between different establishments and different packages. No halogenated VOCs were found in products from only three of the 15 establishments included in the study. PMID- 19188935 TI - [Surveillance of the sanitary conditions of a public swimming pool in the city of Palermo (Italy)]. AB - In a previous study we evaluated the microbiological quality of water of seven pools in the city of Palermo through evaluation of bacterial indicators of faecal contamination and of protozoa (Giardia and Cryptosporidium). In this study we also searched for the presence of fungi in two swimming pools of a public swimming facility in the same city. Samples were collected from both pools, their filtration systems and floor surfaces of the facility. Chemical-physical and microbiological examination of water of the two pools have shown that quality of water depends on the concentration of residual free chlorine and on the number of bathers in the pool. The values of four microbiological parameters (bacterial load at 22 degrees C and 36 degrees C, presence of coagulase-negative Staphylococci and Pseudomonas spp.) increased with diminishing chlorine concentrations and with increasing number of pool users. Faecal bacteria, Giardia and Cryptosporidium were not found. On the other hand, various fungi were isolated from floor surfaces and pool water even in the presence of optimal chlorine concentrations. This study confirms the importance of regular maintenance of pool disinfection systems and suggests the need to search for other micro-organisms not included in the current legislation (Giardia, Cryptosporidium and fungi). PMID- 19188936 TI - [Use of information systems for managing healthcare services in a long-term care facility]. AB - The aim of this study was to evaluate the use of a patient data management information system in a long-term care facility in Brescia (Italy). A survey was conducted among the facility's nursing staff to investigate work organization and identify critical areas that could be improved through the implementation of an information system. A specific patient data management system was then developed and performance indicators were identified. Performance indicators were measured before implementation and then at 6 and 30 months. Managing daily patient healthcare through the use of an information system has allowed an improvement of the appropriateness and quality of care, as well as improved efficiency by saving valuable nursing time. PMID- 19188937 TI - [Prevalence of hearing loss in elderly individuals over 65 years of age: a pilot study in Lombardia (Italy)]. AB - A cross sectional study was performed to determine the prevalence of hearing loss in individuals over age 65 years presenting to a medical centre in Lombardia (Italy). The aim was to assess the feasibility of performing a survey to evaluate the prevalence of hearing loss in the entire region. Audiometric testing was performed in each enrolled subject, to reveal losses in hearing levels at 250 to 8000 Hz; hearing levels were measured in decibels. Prevalences of mild, moderate, severe, and profound hearing impairment were found to be 39%, 17.4%, 2.9% and 0.6% respectively when using the WHO classification, and 59.9%, 25.6%, 2.9%, and 0.6% respectively when using the classification of the Bureau International d'Audiophonologie (BIAP).This study has shown that a relevant part of the study population is affected by hearing impairment and has served as a pilot study for a larger-scale study to be performed in the Lombardia region. PMID- 19188938 TI - [Epidemiology of femoral neck fractures in Puglia (Italy): an analysis of existing data]. AB - The aim of this study was to describe the epidemiology of femoral neck fractures in Puglia (Italy) by analysing hospital discharge records from 1998 to 2005 and mortality data from 1998 to 2003. In total, 41,354 admissions for femoral neck fractures were recorded, 75% of which in females. The mean age of patients was 78 years (73 in males, 79 in females). Approximately 70% of patients below 50 years of age were male, while above age 50, only 23% of patients were male. Yearly admission rates increased from 1998 to 2005. Mortality records recorded 1,031 deaths due to femoral neck fracture with a decreasing trend in mortality rates in the years considered. Fractures in males were most commonly associated with work related accidents and motor vehicle accidents. In females most fractures were related to home accidents. These results may be used to guide prevention strategies. PMID- 19188939 TI - [Actors and processes involved in implementing healthcare policies effectively: theoretical models and case studies]. AB - When implementing healthcare policies it is common to observe a "gap" between policy objectives and results. This because effective translation of healthcare policy into action, from its formulation to final evaluation, requires the contemporary presence of various essential factors and conditions. In this paper, we have attempted to describe and analyse the various existing theoretical models of implementation of healthcare policy and critically discuss them also through the use of specific case studies. PMID- 19188940 TI - [Economic impact of healthcare-associated infections]. AB - Healthcare-associated infections are not only an important public health problem but also have a relevant socioeconomic impact. The overall estimated yearly costs vary between 3.5 billion euros in the United States to 1.3 billion euros in England. In Italy estimated costs are 2.5-5.0 billion euros/year with the cost of a single case ranging from 9,000 to 10,500 euros. The present study aimed to describe the type and distribution of hospital costs for healthcare-associated infections, the economic impact of prevention programs and models for cost effectiveness evaluation of prevention programs. A review of the existing published literature was conducted; studies were included in the review according to whether they had included a control group and to the number of subjects enrolled. Healthcare associated infections lead to increased direct, indirect and intangible costs. Most economic analyses that have been performed consider only direct costs, estimated by calculating the costs of extra days of admission. Surgical site and bloodstream infections were found to be the most costly types of infections, followed by lower respiratory tract and urinary tract infections. There was wide variation in costs between individual hospital departments and countries. Control and prevention strategies have been shown to be effective and efficient but should be evaluated within a specific local context. Prevention programs should therefore take into consideration the local situation. PMID- 19188941 TI - [Influenza: waiting for pandemic]. AB - Pandemic flu is a sudden invasive infection, caused by the diffusion to humans of a viral strain characterized by the presence of one or more new surface proteins. Pandemic influenza prevention is among health priorities for WHO, that introduced a world surveillance programme and published Guidelines for preparing national pandemic plans. Following Guidelines, Italy prepared a plan considering human and veterinary health activities, among which the use of vaccines and antiviral drugs, and informative and formative projects. PMID- 19188942 TI - Phase locking of a 1.5 Terahertz quantum cascade laser and use as a local oscillator in a heterodyne HEB receiver. AB - We demonstrate for the first time the closure of an electronic phase lock loop for a continuous-wave quantum cascade laser (QCL) at 1.5 THz. The QCL is operated in a closed cycle cryo cooler. We achieved a frequency stability of better than 100 Hz, limited by the resolution bandwidth of the spectrum analyser. The PLL electronics make use of the intermediate frequency (IF) obtained from a hot electron bolometer (HEB) which is downconverted to a PLL IF of 125 MHz. The coarse selection of the longitudinal mode and the fine tuning is achieved via the bias voltage of the QCL. Within a QCL cavity mode, the free-running QCL shows frequency fluctuations of about 5 MHz, which the PLL circuit is able to control via the Stark-shift of the QCL gain material. Temperature dependent tuning is shown to be nonlinear, and of the order of -16 MHz/K. Additionally we have used the QCL as local oscillator (LO) to pump an HEB and perform, again for the first time at 1.5 THz, a heterodyne experiment, and obtain a receiver noise temperature of 1741 K. PMID- 19188943 TI - Effect of PDL-induced coherent crosstalk on polarization-division-multiplexed direct-detection systems. AB - We analyze the effect of polarization-dependent loss (PDL) on the performance of the polarization-division-multiplexed (PDM) signals in direct-detection systems. We present the theoretical analysis of the sensitivity penalty caused by PDL induced coherent crosstalk and compare the results with experimental data obtained by using a simple variable PDL emulator based on a LiNbO3 Mach-Zehnder modulator. Also presented is the performance degradation caused by the polarization misalignment between the received signals and the polarization beam splitter at the receiver in the presence of PDL. PMID- 19188944 TI - 2 kW incoherent beam combining of four narrow-linewidth photonic crystal fiber amplifiers. AB - We report on beam combining of four narrow-linewidth fiber amplifier chains, running at different wavelengths and each delivering 500 W optical output power. The main amplifier stage consists of a large mode area photonic crystal fiber. The four output beams of the amplifier chains are spectrally (incoherent) combined using a polarization-independent dielectric reflective diffraction grating to form an output beam of 2 kW continuous-wave optical power with good beam quality (M(2)x = 2.0, M(2)y = 1.8). PMID- 19188945 TI - Enhanced 40 and 80 Gb/s wavelength conversion using a rectangular shaped optical filter for both red and blue spectral slicing. AB - By using a tunable filter with tunability of both bandwidth and wavelength and a very sharp filter roll-off, considerable improvement of all optical Wavelength Conversion, based on Cross Gain and Phase Modulation effects in a Semiconductor Optical Amplifier and spectral slicing, is shown. At 40 Gb/s slicing of blue spectral components is shown to result in a small penalty of 0.7 dB, with a minimal eye broadening, and at 80 Gb/s the low demonstrated 0.5 dB penalty is a dramatic improvement over previously reported wavelength converters using the same principal. Additionally, we give for the first time quantitative results for the case of red spectral slicing at 40 Gb/s which we found to have only 0.5 dB penalty and a narrower time response, as anticipated by previously published theoretical papers. Numerical simulations for the dependence of the eye opening on the filter characteristics highlight the importance of the combination of a sharp filter roll-off and a broad passband. PMID- 19188946 TI - Layer-to-layer optical interconnect coupling by soft-lithographic stamping. AB - We designed and developed a fabrication method for a polymeric waveguide connecting multiple optical interconnect layers in three-dimensional integrated electro-optical systems, using a series of silicone elastomer stamps. The scalable process is a deterministic printing method, generating optical S-shaped polymeric lightwave coupling for interconnects among separate layers. General characteristics of S-shaped coupling among layers with various separation distances were investigated too. The soft-lithography fabrication process of this coupling interconnection design allows for significant advantages over traditional designs and fabrication methods in terms of insertion loss as well as 3D integration capability, when used in high-bandwidth printed circuit boards. PMID- 19188947 TI - 600 W power scalable single transverse mode tapered double-clad fiber laser. AB - Pump propagation and absorption in active tapered double-clad fiber has been analyzed based on a ray optics approach. Optimization of the longitudinal shape, absorption and angular distribution of the pump beam allowed for power scaling of a ytterbium fiber laser up to 600 W with high beam quality (M2300 nm at a repetition rate of 100 kHz. The pulses have been compressed to 92 fs by a 4-prism compressor. PMID- 19188963 TI - Monopole woodpile photonic crystal modes for light-matter interaction and optical trapping. AB - Two types of ultra-high-Q monopole modes are designed in a woodpile three dimensional photonic crystal. The unit cell size modulation is applied to a woodpile photonic crystal waveguide in a complete photonic band gap. A monopole mode overlapping with a dielectric rod is designed for solid-state sub-wavelength scale light-matter interaction devices such as nanolasers, cavity-QED and optical switches, whereas another type of monopole mode overlapping with vacuum is designed for optical trapping experiments. For the mode overlapping with vacuum, the mode volume is as small as 0.4 cubic half-wavelengths. PMID- 19188964 TI - Simultaneous measurement method of total and self-interference for the volumetric thickness-profilometer. AB - A simultaneous measurement method for the total interference and self interference of a sample is proposed. The proposed method is capable of making separate measurements of the thickness and surface profile of micro-patterned thin film. The system is an extension of a full-field wavelength scanning interferometer with a single acousto-optic tunable filter (AOTF) as a spectral imaging device. Separate measurements are realized via the polarization-sensitive diffraction of non-collinear acousto-optic interaction. That is, the diffraction angle of an AOTF is separated into different directions depending on the polarization state of the incident light. In so doing, the polarization states of a reference and a sample light were manipulated differently so that a single AOTF can generate the total interference and the self-interference signal in different directions simultaneously. Thus, a compact and light-efficient system is realized with an AOTF, a beam splitter and two CCDs. Thus, a compact system with light efficiency of two to four times higher than the previously reported system is realized with an AOTF, a beam splitter and two CCDs. Details of the calibration procedures such as wavelength-frequency relation, image shift and registration between two CCDs are provided for the proposed setup. Experimental results are provided and compared to those using commercial equipment that demonstrate the efficiency of the proposed system for the high-speed measurements of the thickness and the surface profile of micro-patterned thin film. PMID- 19188965 TI - Single cellgap transflective liquid crystal cell with high contrast and high cellgap tolerance. AB - In this paper an optical configuration of a transflective liquid crystal (LC) cell driven by a horizontal electric field is proposed, which shows high contrast, high cellgap tolerance, and single gamma, simultaneously. The dark state of the reflective part is realized by a polarizer (0 degrees), a half-wave plate (15 degrees), LC layer (120 degrees), and a quarter-wave layer (-15 degrees), while a wide-band quarter-wave plate (45 degrees) and a polarizer (90 degrees) are added for the dark state of the transmissive part. Since the optic axis of the homogeneously aligned LC layer is set to be parallel to the polarization direction of the light passed through the half-wave plate, the dark state is rarely affected by the cellgap of the LC layer. Due to the different directions of the electric fields, LCs are rotated to 97.5 degrees for the bright state of the reflective part, but to 75 degrees for that of the transmissive part. With the proposed configuration, a high contrast single-gamma transflective display with high cellgap tolerance can be realized in a single-cellgap structure. PMID- 19188966 TI - A strategy for quantitative spectral imaging of tissue absorption and scattering using light emitting diodes and photodiodes. AB - A diffuse reflectance spectroscopy system was modified as a step towards miniaturization and spectral imaging of tissue absorption and scattering. The modified system uses a tunable source and an optical fiber for illumination and a photodiode in contact with tissue for detection. Compared to the previous system, it is smaller, less costly, and has comparable performance in extracting optical properties in tissue phantoms. Wavelength reduction simulations show the feasibility of replacing the source with LEDs to further decrease system size and cost. Simulated crosstalk analysis indicates that this evolving system can be multiplexed for spectral imaging in the future. PMID- 19188967 TI - Evaluation of the computational effort for chromatic dispersion compensation in coherent optical PM-OFDM and PM-QAM systems. AB - Recently, coherent-detection (CoD) polarization multiplexed (PM) transmission has attracted considerable interest, specifically as a possible solution for next generation systems transmitting 100 Gb/s per channel and beyond. In this context, enabled by progress in ultra-fast digital signal processing (DSP) electronics, both multilevel phase/amplitude modulated formats (such as QAM) and orthogonal frequency-division multiplexed (OFDM) formats have been proposed. One specific feature of DSP-supported CoD is the possibility of dealing with fiber chromatic dispersion (CD) electronically, either by post-filtering (PM-QAM) or by appropriately introducing symbol-duration redundancy (PM-OFDM). In both cases, ultra-long-haul fully uncompensated links seem to be possible. In this paper we estimate the computational effort required by CD compensation, when using the PM QAM or PM-OFDM formats. Such effort, when expressed as number of operations per received bit, was found to be logarithmic with respect to link length, bit rate and fiber dispersion, for both classes of systems. We also found that PM-OFDM may have some advantage over PM-QAM, depending mostly on the over-sampling needed by the two systems. Asymptotically, for large channel memory and small over sampling, the two systems tend to require the same CD-compensation computational effort. We also showed that the effort required by the mitigation of polarization related effects can in principle be made small as compared to that of CD over long uncompensated links. PMID- 19188968 TI - The role of input chirp on phase shifters based on slow and fast light effects in semiconductor optical amplifiers. AB - We experimentally investigate the initial chirp dependence of slow and fast light effects in a semiconductor optical amplifier followed by an optical filter. It is shown that the enhancement of the phase shift due to optical filtering strongly depends on the chirp of the input optical signal. We demonstrate approximately 120 degrees phase delay as well as approximately 170 degrees phase advance at a microwave frequency of 19 GHz for different optimum values of the input chirp. The experimental results are shown to be in good agreement with numerical results based on a four-wave mixing model. Finally, a simple physical explanation based on an analytical perturbative approach is presented. PMID- 19188969 TI - Well-chosen method for an optimal design of doublet lens design. AB - This paper presents a method for choosing a doublet design for the correction of longitudinal chromatic, spherical and coma aberrations. A secondary dispersion formula is utilized to sort out minimal longitudinal chromatic aberrations for the doublet. The program is developed with the Matlab software. An optimal doublet design to efficiently reduce both spherical aberration and coma will incorporate glass combination with a sufficiently large difference in the V numbers and small powers. We succeed in obtaining an optimal doublet design with the proposed method. PMID- 19188970 TI - A saturable absorber Q-switched all-fiber ring laser. AB - We propose a simple design of a saturable absorber Q-switched all-fiber ring laser. By locating a saturable absorber fiber in the intensity-enhanced section of a ring resonator, the laser is passively Q-switched. A set of location dependent rate equations is established for Q-switching modeling. The design has been numerically and experimentally demonstrated using Er(3+)-doped fiber at the emission wavelength of 1550 nm. A single-mode Q-switched pulse with pulse energy of 0.37 microJ and pulse duration of 218 ns was achieved with 980-nm pump power near 7 mW. PMID- 19188971 TI - Unrepeated 200-km transmission of 40-Gbit/s 16-QAM signals using digital coherent receiver. AB - We demonstrate unrepeated 200-km transmission of 40-Gbit/s 16-QAM signals using a digital coherent receiver, where the decision-directed carrier-phase estimation is employed. The phase fluctuation is effectively eliminated in the 16-QAM system with such a phase-estimation method, when the linewidth of semiconductor lasers for the transmitter and the local oscillator is 150 kHz. Finite-impulse-response (FIR) filters at the receiver compensate for 4,000-ps/nm group-velocity dispersion (GVD) of the 200-km-long single-mode fiber and a part of self-phase modulation (SPM) in the digital domain. In spite of the launched power limitation due to SPM, the acceptable bit-error rate performance is obtained owing to high sensitivity of the digital coherent receiver. PMID- 19188973 TI - Photonic crystal fiber interferometer for chemical vapor detection with high sensitivity. AB - We report an in-reflection photonic crystal fiber (PCF) interferometer which exhibits high sensitivity to different volatile organic compounds (VOCs), without the need of any permeable material. The interferometer is compact, robust, and consists of a stub of PCF spliced to standard optical fiber. In the splice the voids of the PCF are fully collapsed, thus allowing the excitation and recombination of two core modes. The device reflection spectrum exhibits sinusoidal interference pattern which shifts differently when the voids of the PCF are infiltrated with VOC molecules. The volume of voids responsible for the shift is less than 600 picoliters whereas the detectable levels are in the nanomole range. PMID- 19188972 TI - High speed phase shifting interferometry using injection locking of the laser frequency to the resonant modes of a confocal Fabry-Perot cavity. AB - We present a high speed phase shifting interferometer which utilizes the self injection locking of a frequency tunable laser diode. By using a confocal Fabry Perot cavity made of ultra low expansion glass, and linearly modulating the laser diode current, the laser frequency could be injection locked to the resonant modes of the Fabry-Perot cavity consecutively. It provided equal phase steps to the interferograms which are ideal to be analyzed by the Carre algorithm. The phase step error was evaluated to be about 3 MHz which corresponds to 0.2 nm in length measurement. With this technique, profile measurements are insensitive to external vibration since four 640x480 pixels images can be acquired within 4 ms. Difference of two profile measurements, each made with and without vibration isolation, respectively, was evaluated to be 0.5 nm (rms). PMID- 19188974 TI - Method for effectively utilizing tunable one-pump fiber parametric wavelength converters as an enabling device for WDM routers. AB - In this paper a method is proposed to maximize the bandwidth of the WDM router based on one-pump fiber parametric wavelength converters. It is proved that for such converters there exists an optimum signal (idler) frequency at which the output (input) tuning range can be maximized. Analytical expressions of the optimum frequency and the maximal tuning range are deduced. Then a two-stage bidirectional wavelength conversion method is proposed. With this method the bandwidth of the WDM router based on such a converter can be significantly improved compared to the one-stage ones by 252% if ordinary highly nonlinear fibers are used or 390% if fibers with optimal fourth order dispersion are used. PMID- 19188975 TI - Rigorous electromagnetic analysis of two dimensional micro-axicon by boundary integral equations. AB - The focal performance of the micro-axicon and the Fresnel axicon (fraxicon) are investigated, for the first time, by the rigorous electromagnetic theory and boundary element method. The micro-axicon with different angle of apex and the fraxicon with various period and angle of apex are investigated. The dark segments of the fraxicon are explored numerically. Rigorous results of focal performance of the micro-axicon and the fraxicon are different from the results given by the approximation of geometrical optics and the scalar diffraction theory. The scattering effects are dominant in the fraxicon with small size of feature. It is expected that our study can provides very useful information in analyzing the axicon in optical trapping systems. PMID- 19188976 TI - Tuning the frequency of few-cycle femtosecond laser pulses by molecular phase modulation. AB - We demonstrate theoretically how the time-dependent phase modulation induced by molecular alignment can be used to tune the frequency of few-cycle femtosecond laser pulses continuously. Using impulsively excited alignment in N(2), the central wavelength of an initial 800 nm, 5 fs Gaussian pulse can be tuned from 324.6 to 4237.3 nm. The aligned N(2) molecules are obtained by pretransmitting another 800 nm, 100 fs linearly polarized laser pulse of intensity 3.5x10(13) Wcm(-2). The number of optical cycles contained in the frequency-tuned pulse is almost unchanged after ideal chirp compensation. PMID- 19188977 TI - Application of Hertz vector diffraction theory to the diffraction of focused Gaussian beams and calculations of focal parameters. AB - Hertz vector diffraction theory is applied to a focused TEM00 Gaussian light field passing through a circular aperture. The resulting theoretical vector field model reproduces plane-wave diffractive behavior for severely clipped beams, expected Gaussian beam behavior for unperturbed focused Gaussian beams as well as unique diffracted-Gaussian behavior between the two regimes. The maximum intensity obtainable and the width of the beam in the focal plane are investigated as a function of the clipping ratio between the aperture radius and the beam width in the aperture plane. PMID- 19188978 TI - Monitoring the He+ ion channel formation by high-order harmonic generation. AB - The macroscopic build-up of the high-order harmonic signal depends on the free electron density in the generation medium. The free electrons affect the harmonic yield and spectral shape through modifying the refractive index and the phase matching conditions. These dependences allow studying the He(+) ion channel formation in a He gas jet. The evolution of an ion channel created by an ultrashort laser pulse via optical field ionization was monitored using the harmonic signal generated by a collinear propagating second laser pulse. From the measured high harmonic signal as function of the delay we are able to gain information about the free electron density. Under our experimental condition, the ion channel has been fully formed 300 fs after the first laser pulse, resulting in an enhancement of harmonic yield of the second laser pulse by two orders of magnitude. PMID- 19188979 TI - Blue solitary waves from infrared continuous wave pumping of optical fibers. AB - Modulation instability of a continuous infrared wave leading to red-shifting solitons and blue-shifting dispersive waves and then subsequent soliton trapping is shown to enable the generation of blue solitary waves in optical fibers. The physical mechanism is described in the context of continuous wave supercontinuum generation leading to a spectrum which spans the visible and near infrared within practical experimental conditions. PMID- 19188980 TI - A novel ultra-low loss hollow-core waveguide using subwavelength high-contrast gratings. AB - We propose a novel ultra-low loss single-mode hollow-core waveguide using subwavelength high-contrast grating (HCG). We analyzed and simulated the propagation loss of the waveguide and show it can be as low as 0.006 dB/m, three orders of magnitude lower than the lowest loss of the state-of-art chip-scale hollow waveguides. This novel HCG hollow-core waveguide design will serve as a basic building block in many chip-scale integrated photonic circuits enabling system-level applications including optical interconnects, optical delay lines, and optical sensors. PMID- 19188981 TI - Ultra-high sensitivity optical stress sensor based on double-layered photonic crystal microcavity. AB - In this report, we present the design principles to achieve a highly sensitive optical stress sensor. The structure we use is a double-layered (DL) photonic molecule with optical bonding and anti-bonding states based on whispering-gallery mode in photonic crystal microcavity. By applying finite-difference time-domain and finite-element methods, we simulate the change of optical properties (including wavelength and quality (Q) factor) of bonding mode caused by the DL structural variation due to the applied stress in two DL geometries. In the end, we summarize an optical stress sensor design with high Q factor, large structural response due to the applied stress, and large optical spectrum change due to the DL structural variation. The minimum detectable stress variation is estimated to be as small as 0.95 nN. PMID- 19188982 TI - Slot waveguides with polycrystalline silicon for electrical injection. AB - We demonstrate horizontal slot waveguides using high-index layers of polycrystalline and single crystalline silicon separated by a 10 nm layer of silicon dioxide. We measure waveguide propagation loss of 7 dB/cm and a ring resonator intrinsic quality factor of 83,000. The electric field of the optical mode is strongly enhanced in the low-index oxide layer, which can be used to induce a strong modal gain when an active material is embedded in the slot. Both high-index layers are made of electrically conductive silicon which can efficiently transport charge to the slot region. The incorporation of conductive silicon materials with high-Q slot waveguide cavities is a key step for realizing electrical tunneling devices such as electrically pumped silicon-based light sources. PMID- 19188983 TI - Acceptance angle effects on the beam attenuation in the ocean. AB - The beam attenuation serves as a proxy for particulate matter and is a key parameter in visibility algorithms for the aquatic environment. It is well known, however, that the beam attenuation is a function of the acceptance angle of the transmissometer used to measure it. Here we compare eight different transmissometers with four different acceptance angles using four different deployment strategies and sites, and find that their mean attenuation values differ markedly and in a consistent way with instrument acceptance angle: smaller acceptance angles provide higher beam attenuation values. This difference is due to variations in scattered light collected with different acceptance angles and is neither constant nor easy to parameterize. Variability (in space or time) in the ratios of beam attenuations measured by two different instruments correlates, in most cases, with the particle size parameter (as expected from Mie theory), but this correlation is often weak and can be the opposite of expectations based on particle size changes. We recommended careful consideration of acceptance angle in applications of beam transmission data especially when comparing data from different instruments. PMID- 19188984 TI - Subnanosecond mJ eye-safe laser with an intracavity optical parametric oscillator in a shared resonator. AB - We theoretically verify that the threshold of an intracavity optical parametric oscillator pumped by a passively Q-switched laser is entirely controlled by the bleach of the saturable absorber not by the signal output reflectivity. We use a series of different output couplers to optimize the output performance. With a signal output reflectivity of 15%, we experimentally achieve an efficient subnanosecond eye-safe laser with 3.3 mJ pulse energy and 1.5 MW peak power. PMID- 19188985 TI - mhFLIM: resolution of heterogeneous fluorescence decays in widefield lifetime microscopy. AB - Frequency-domain fluorescence lifetime imaging microscopy (FD-FLIM) is a fast and accurate way of measuring fluorescence lifetimes in widefield microscopy. However, the resolution of multiple exponential fluorescence decays has remained beyond the reach of most practical FD-FLIM systems. In this paper we describe the implementation of FD-FLIM using a 40 MHz pulse train derived from a supercontinuum source for excitation. The technique, which we term multi-harmonic FLIM (mhFLIM), makes it possible to accurately resolve biexponential decays of fluorophores without any a priori information. The system's performance is demonstrated using a mixture of spectrally similar dyes of known composition and also on a multiply-labeled biological sample. The results are compared to those obtained from time correlated single photon counting (TCSPC) microscopy and a good level of agreement is achieved. We also demonstrate the first practical application of an algorithm derived by G. Weber [1] for analysing mhFLIM data. Because it does not require nonlinear minimisation, it offers potential for realtime analysis during acquisition. PMID- 19188986 TI - Terahertz Bessel-Gauss beams of radial and azimuthal polarization from microstructured photoconductive antennas. AB - We report on emission and detection of pulsed terahertz radiation of radial and azimuthal polarization by microstructured photoconductive antennas. To this end the electrode geometry of the emitter is inverse to the desired THz field pattern and a second periodic structure prevents destructive interference effects. Beam profiles of freely propagating THz waves are studied for divergent and refocused beams. They can be well described as the lowest order Bessel-Gauss modes with a divergence comparable to linearly polarized Gaussian beams. Additionally, mode sensitive detection is demonstrated for radially polarized radiation. PMID- 19188987 TI - White organic light-emitting diodes combining vacuum deposited blue electrophosphorescent devices with red surface color conversion layers. AB - We report white organic light-emitting diodes (WOLEDs) combining vacuum deposited blue electrophosphorescent devices with red surface color conversion layers (CCLs). With an iridium (III) [bis(4,6-di-fluoropheny)- pyridinato-N,C(2')] picolinate (FIrpic) doped 4,4'-bis(9-carbazolyl)-2,2'-dimethyl-biphenyl (CDBP) blue electrophosphorescent light emitting layer, and an appropriate red surface CCL containing 4-(dicyanomethylene)-2-tert-butyl-6-(1,1,7,7-tetramethyljulolidyl 9-enyl)-4H-pyran (DCJTB), the WOLED generate high efficiency and very pure white light with a peak luminous (power) efficiency of 18.1 cd/A (9.5 lm/W) and CIE coordinates of (0.32, 0.31), very close to the equal-energy white, respectively. Moreover, the output spectra and CIE coordinates of the WOLED show no significant change at a wide range of current density. PMID- 19188988 TI - Numerically-assisted coupled-mode theory for silicon waveguide couplers and arrayed waveguides. AB - We investigate coupled-mode theory in designing high index contrast silicon-on insulator waveguide couplers and arrayed waveguides. We develop and demonstrate a method of solution to the inverse problem of reconstructing the coupling matrix from the modal profiles obtained, in this case, from finite-difference frequency domain field calculations. We show that whereas supermode theory provides a good approximation of the mode profiles, next-to-nearest-neighbor coupling becomes significant at small separation distances between arrayed waveguides. These distances are quantified for three different silicon-on-insulator material platforms. We also point out the phenomenon of field skewing and deformation at small separations. PMID- 19188989 TI - Mueller decomposition images for cervical tissue: potential for discriminating normal and dysplastic states. AB - We report the potential of Mueller decomposition images to discriminate the normal against the dysplastic (precancerous) states in cervical tissue. It is observed that in the epithelium region, depolarization power is sensitive to morphological changes during progression from normal to dysplastic state while retardance and diattenuation do not show any significant change. These morphological changes have been correlated with the microscopic images of the tissues. In contrast, it is the retardance which reveals the morphological changes around the stromal region. Additionally, we have evaluated the arithmetic mean of depolarization power and retardance from their respective decomposed images and have shown that this parameter has a potential to discriminate normal tissues against dysplastic ones. PMID- 19188990 TI - Spectral determination of a two-parametric phase function for polydispersive scattering liquids. AB - A method for determining a two-parametric Gegenbauer-kernel phase function that accurately describes the diffuse reflectance from a polydispersive scattering media at small source-detector separations (0.23 to 1.2 mm), is presented. The method involves spectral collimated transmission measurements, spatially resolved spectral diffuse reflectance (SRDR) measurements, and inverse Monte Carlo technique. Both absolute calibration (using a monodispersive polystyrene microsphere suspension) and relative calibration (eliminating differences between fibers) of SRDR spectra yielded comparable results. When applied to water dilutions of milk, simulated and measured spectra deviated less than 6.5% and 2.5% for the absolute and relative calibration case, respectively, even for the closest fiber separation. Corresponding values for milk including ink as an absorber, were 13.4% and 7.3%. PMID- 19188991 TI - The possibility for surface plasmons lasers. AB - We have demonstrated for the first time, attributes of a surface plasmons' laser: threshold, gain, spectral line narrowing and feedback in the visible range. The surface metallic waveguides were consisted of a nano-scale hole-array in a 50 nm thick layer of aluminum oxide on top of aluminum substrate (anodized aluminum oxide or, AAO). In some cases, two-layer graphene was added on top of the perforated oxide layer, as well. The sub-wavelength array of holes enabled coupling to and from the waveguides as well as, providing feedback to the surface modes. The gain media molecules (fluorescein) were imbedded in the structure's pores. Threshold and spectral line narrowing of 30% were clearly demonstrated when pumped with a pulsed laser. PMID- 19188992 TI - Dispersion engineering of slow light photonic crystal waveguides using microfluidic infiltration. AB - We present a technique based on the selective liquid infiltration of photonic crystal (PhC) waveguides to produce very small dispersion slow light over a substantial bandwidth. We numerically demonstrate that this approach allows one to control the group velocity (from c/20 to c/110) from a single PhC waveguide design, simply by choosing the index of the liquid to infiltrate. In addition, we show that this method is tolerant to deviations in the PhC parameters such as the hole size, which relaxes the constraint on the PhC fabrication accuracy as compared to previous structural-based methods for slow light dispersion engineering. PMID- 19188993 TI - Extraordinary transmittance in three dimensional crater, pyramid, and hole-array structures prepared through reversal imprinting of metal films. AB - We used a reversal imprinting-in-metal (RIM) process to fabricate various three dimensional (3D) metal structures under low pressure. Molds featuring different shapes were used to pattern various subwavelength metal structures, including pyramidal, hole-array, and crater-like structures. Refractive index matching and cavity effects both enhanced the degree of transmission of these structured metal films. The crater-like structure appears to be a promising material because of the unique properties imparted by the elongated and gradually tapering spacing of its cavities. From both near-field simulations and experimentally obtained optical spectra, we found that the cavity effect in the crater-like structure led to significantly enhanced transmission of the optical intensity. Thus, this RIM process allows the ready fabrication of various two- and three-dimensional metallic structures for use in surface plasmon-based devices. PMID- 19188994 TI - Optical waves in partially ordered anisotropic media. AB - This paper investigates light wave propagation in partially ordered birefringent optical media, with focusing on the influence of the degree of molecular ordering on the propagating modes. The double refraction at isotropic/partially ordered interface is also studied, where the impact of the order parameter on the ordinary and extraordinary refracted waves is discussed. This paper also demonstrates the importance of involving the order parameter in studying the total internal reflection (TIR). PMID- 19188995 TI - Doppler-free spectroscopy of molecular iodine using a frequency-stable light source at 578 nm. AB - A stable light source obtained using sum-frequency generation (SFG) is developed for high-resolution spectroscopy at 578 nm. Hyperfine transitions of molecular iodine are observed by using the SFG light source with saturation spectroscopy. The light source is frequency stabilized to the observed hyperfine transition and achieves a stability of 2x10(-12) for a 1-s averaging time. The absolute frequency of the light source stabilized on the a1 component of the R(37)16-1 transition is determined as 518304551833 (2) kHz. This transition serves as a frequency reference for the (1)S(0)-(3)P(0) optical clock transition in neutral ytterbium (Yb). PMID- 19188996 TI - All-fiber devices based on photonic crystal fibers with integrated electrodes. AB - A special kind of microstructured optical fiber is proposed and fabricated in which, in addition to the holey region (solid core and silica-air cladding), two large holes exist for electrode insertion. Either Bi-Sn or Au- Sn alloys were selectively inserted into the large holes forming two parallel, continuous and homogeneous internal electrodes. We demonstrate the production of a monolithic device and its use to externally control some of the guidance properties (e.g. polarization) of the fiber. PMID- 19188997 TI - Continuous-wave and Q-switched operation of a compact, diode-pumped Yb3+:KY(WO4)2 planar waveguide laser. AB - A diode-pumped LPE-grown Yb:KYW planar waveguide laser is demonstrated in a microchip monolithic cavity configuration. Output powers as high as 148 mW and thresholds as low as 40 mW were demonstrated during continuous-wave operation. Pulses of 170 ns duration with maximum pulse energy of 44 nJ at a 722 kHz repetition rate were generated when Q-switched using a semiconductor saturable absorber mirror. PMID- 19188998 TI - Estimating scattering of pure water from density fluctuation of the refractive index. AB - The use of density derivative of the refractive index from the classic Lorentz Lorenz equation or its variations performed poorly in estimating the scattering by water, leading to the alternative use of pressure derivative instead, which however has been scarcely measured due to its extremely low sensitivity. Recently, density derivative has been deduced directly from theoretical models. Three characterizations of density derivative of the refractive index were evaluated and scattering of water thus calculated converge with each other within 3.5% and agree with the measurement by Morel (Cahiers Oceanographiques, 20, 157, 1968) within 2% (with depolarization ratio = 0.039), all improving over the earlier estimates based on either density or pressure derivatives. Taking into account of uncertainty associated with the depolarization ratio, the prediction based on the model by Proutiere et al. (J. Phy. Chem., 96, 3485, 1992) still agrees with the measurement within the experimental errors (2%). PMID- 19188999 TI - Characteristics of dielectric-band modified single-cell photonic crystal lasers. AB - We demonstrate new types of dielectric-band photonic crystal lasers in a two dimensional modified single-cell cavity with enlarged air holes. Finite difference time-domain simulations performed in real and Fourier spaces show that the dielectric-band cavity modes originating from the first band edge point in the dielectric band have mode patterns that are distinguishable from conventional air-band cavity modes. In our experiment, the observed multimode lasing peaks are identified as the hexapole and the monopole dielectric-band cavity modes through the spectral positions and mode images. The thresholds of these lasers are measured as approximately 340 microW and approximately 450 microW, respectively, at room temperature. In addition, using the simulation based on the actual fabricated structures, quality factors and mode volumes are computed as 4900 and 1.09 (lambda/n)3 for the hexapole mode, and 4300 and 2.27 (lambda/n)3 for the monopole mode, respectively. PMID- 19189000 TI - Optically multiplexed imaging with superposition space tracking. AB - We describe a novel method to track targets in a large field of view. This method simultaneously images multiple, encoded sub-fields of view onto a common focal plane. Sub-field encoding enables target tracking by creating a unique connection between target characteristics in superposition space and the target's true position in real space. This is accomplished without reconstructing a conventional image of the large field of view. Potential encoding schemes include spatial shift, rotation, and magnification. We discuss each of these encoding schemes, but the main emphasis of the paper and all examples are based on one dimensional spatial shift encoding. System performance is evaluated in terms of two criteria: average decoding time and probability of decoding error. We study these performance criteria as a function of resolution in the encoding scheme and signal-to-noise ratio. Finally, we include simulation and experimental results demonstrating our novel tracking method. PMID- 19189001 TI - Investigation of the influence of the aberration induced by a plane interface on STED microscopy. AB - The structure of the inhibition patterns is important to the stimulated emission depletion (STED) microscopy. Usually, Laguerre- Gaussian (LG) beam and the central zero-intensity patterns created by inserting phase masks in Gaussian beams are used as the erase beam in STED microscopy. Aberration is generated when focusing beams through an interface between the media of the mismatched refractive indices. By use of the vectorial integral, the effects of such aberration on the shape of depletion patterns and the size of fluorescence emission spot in the STED microscopy are studied. Results are presented as a comparison between the aberration-free case and the aberrated cases. PMID- 19189002 TI - WDM up-conversion employing frequency quadrupling in optical modulator. AB - This work presents an optical up-conversion system with frequency quadrupling for wavelength-division-multiplexing (WDM) communication systems using a dual parallel Mach-Zehnder modulator without optical filtering. Four-channel 1.25-Gb/s wired fiber-to-the-x (FTTx) and wireless radio-over-fiber (RoF) signals are generated and transmitted simultaneously. Moreover, the decline in receiver sensitivities due to Mach-Zehnder modulator bias drifts is also investigated. Receiver power penalties of the 20-GHz up-converted WDM signals and baseband (BB) FTTx signals are less than 1 dB when bias deviation voltage is less the 20% of the half-wave voltage. After transmission over a 50-km SSMF, the receiver power penalties of both the BB and 20-GHz RF OOK signals are less than 1 dB. Notably, 60-GHz optical up-conversion can be achieved using 15-GHz radio frequency (RF) components and equipment. PMID- 19189003 TI - Real-time complex temporal response measurements of ultrahigh-speed optical modulators. AB - We demonstrate a technique for direct, real-time characterization of the complex (amplitude and phase) temporal response of ultrahigh-speed (GHz-bandwidth) optical modulators. The demonstrated technique is based on pulse interferometry combined with time-frequency mapping processes using fiber linear dispersion. A new mechanism is incorporated to overcome the temporal resolution (bandwidth) limitation of the detectable modulation response in our previously reported setup. This mechanism, referred to as 'common-path temporal image magnification', lowers the required detection bandwidth by a factor of more than 10, enabling real-time single-shot waveform acquisition without loss of information using a conventional temporal digitizer. The design specifications of the proposed measurement setup are derived and discussed in detail. As a proof-of-concept experiment, real-time characterization of a complex electro-op c modulation temporal response with time features as fast as approximately 35 ps (modulation bandwidth > 40-GHz) was obtained and displayed at a video rate of 30 frames/sec. PMID- 19189004 TI - The van Cittert-Zernike theorem for electromagnetic fields. AB - The van Cittert-Zernike theorem, well known for the scalar optical fields, is generalized for the case of vector electromagnetic fields. The deduced theorem shows that the degree of coherence of the electromagnetic field produced by the completely incoherent vector source increases on propagation whereas the degree of polarization remains unchanged. The possible application of the deduced theorem is illustrated by an example of optical simulation of partially coherent and partially polarized secondary source with the controlled statistical properties. PMID- 19189005 TI - Radiation force of coherent and partially coherent flat-topped beams on a Rayleigh particle. AB - Propagations of coherent and partially coherent flat-topped beams through a focusing optical system are formulated. The radiation force on a Rayleigh dielectric sphere induced by focused coherent and partially coherent flat-topped beams is investigated theoretically. It is found that we can increase the transverse trapping range at the planes near the focal plane by increasing the flatness (i.e., beam order) of the flat-topped beam, and increase the transverse and longitudinal trapping ranges at the focal plane by decreasing the initial coherence of the flat-topped beam. Moreover the trapping stiffness of flat-topped beam becomes lower as the beam order increases or the initial coherence decreases. The trapping stability is also analyzed. PMID- 19189006 TI - Reduction of timing jitter in a Q-Switched Nd:YAG laser by direct bleaching of a Cr4+:YAG saturable absorber. AB - A method for optical triggering of a Q-switched Nd:YAG laser by direct bleaching of a Cr:YAG saturable absorber is described. This method involves the bleaching of a thin sheet of the saturable absorber from a direction orthogonal to the lasing axis using a single laser diode bar, where the Cr:YAG transmission increased from a non-bleached value of 47% to a bleached value of 63%. For steady state operation of a passively Q-switched laser (PRF=10 Hz), the pulse-to-pulse timing jitter showed approximately 12X reduction in standard deviation, from 241 nsec for free running operation to 20 nsec with optical triggering. PMID- 19189007 TI - Metal-free scanning optical microscopy with a fractal fiber probe. AB - Scanning Near-field Optical Microscopy (SNOM) is the leading instrument used to image optical fields on the nanometer scale. A metal-coating is typically applied to SNOM probes to define a subwavelength aperture and minimize optical leakage, but the presence of such coatings in the near field of the sample can often cause a substantial change in the sample emission properties. For the first time, the authors demonstrate near-field imaging on a metal substrate with a metal-free probe made from a novel structured optical fiber, designed to maximize optical throughput and potentially remove the need for the metal. PMID- 19189008 TI - Detuned slow light in the Doppler broadened multi-level D2 line of Rubidium. AB - We observed a detuned slow light phenomenon based on electromagnetically induced transparency in (87)Rb D2 line composed of multiple excited-hyperfine states within a Doppler-broadened linewidth. The results show that the maximum group delay of a probe occurs at off-detuned two-photon resonance frequency. The observed detuned group delay is analyzed with numerical calculations for a probe pulse interacting with the neighboring excited-states-modified Doppler broadening atoms for a fixed coupling field. The experimental results are in good agreement with the numerical calculations. PMID- 19189009 TI - Large temperature sensitivity of Sagnac loop interferometer based on the birefringent holey fiber filled with metal indium. AB - The large temperature sensitivity of the Sagnac loop interferometer based on the birefringent holey fiber filled with metal indium was experimentally demonstrated. The temperature sensitivities of the wavelength shift of the interferometer and the birefringence the fiber with indium were measured to be 6.3 nm/K and -3.3x10(-6) /K, respectively. The large temperature sensitivity of the fiber was explained by introduction of the fiber birefringence change originated from the large thermal expansion property of the metal indium at the elevated temperature. PMID- 19189010 TI - Ultrashort pulse polarization control in silicon waveguides. AB - The nonlinear polarization dynamics of ultrashort optical pulses propagating in a low birefringent silicon waveguide is theocratically and numerically studied, with a static electric field applied across the waveguide. It is shown that the pulse shape and polarization evolution can be efficiently controlled by adjusting the magnitude of the applied dc field. It is also demonstrated that the polarization instability regime can be achieved in such waveguides - despite the presence of strong linear losses - by appropriately engineering the spatial distribution of the control field along the waveguide. The simulations indicate that short silicon waveguides can serve as a viable platform for developing re configurable all-optical and/or optically assisted electro-optic devices in the spectral range spanning from near- to mid-infrared. PMID- 19189012 TI - Polarization insensitive imaging through polarization gratings. AB - Liquid crystal polarization gratings exhibit high diffraction efficiency (approximately 100%) in thin material layers comparable to the radiation wavelength. We demonstrate that they can be combined for polarization-insensitive imaging and optical switching applications. A pair of closely spaced, parallel oriented, cycloidal polarization gratings is capable of canceling the diffractive property of an individual grating. As a result, the phase of the beam is not distorted, and holographic images can be formed through them. An anti-parallel arrangement results in a broader effective diffraction band and doubles the diffraction angle. Broadband diffraction spanning from 480 nm to beyond 900 nm wavelengths has been obtained for a pair of gratings with 500 nm and 633 nm peak diffraction wavelengths. Liquid crystal polymer cycloidal gratings were used in the study showing 98% diffraction efficiency over a large area, and allowed for the use of laser beams expanded to 25 mm. The characteristics of combined cycloidal gratings were tested with laser beams at both UV and red wavelengths. PMID- 19189011 TI - Theoretical investigation of the transverse optical force between a silicon nanowire waveguide and a substrate. AB - We present a study of transverse optical forces arising in a free-standing silicon nanowire waveguide. A theoretical framework is provided for the calculation of the optical forces existing between a waveguide and a dielectric substrate. The force is evaluated using a numerical procedure based on finite element simulations. In addition, an analytical formalism is developed which allows for a simple approximate analysis of the problem. We find that in this configuration optical forces on the order of pN can be obtained, sufficient to actuate nano-mechanical devices. PMID- 19189013 TI - Applications of digital micro-mirror devices to digital optical microscope dynamic range enhancement. AB - In this paper, we present a method of using digital micro-mirror devices to dynamic range enhancement of a digital optical microscope images. Our adaptive feedback illumination control generates a high dynamic range image through an algorithm that combines the DMD-to-camera pixel geometrical mapping and a feedback operation. The feedback process automatically generates an illumination pattern in an iterative fashion that spatially modulates the DMD array elements on pixel-by-pixel level. Via experiment, we demonstrate a system that uses precise DMD control of the projector to enhance the dynamic range ideally by a factor of 573. Results are presented showing approximately 5 times the camera dynamic range, enabling visualization over a wide range of specimen characteristics. PMID- 19189014 TI - Multiple scattering of light in three-dimensional photonic quasicrystals. AB - Recent experiments on three-dimensional icosahedral dielectric photonic quasicrystals have shown several unexpected features: transmitted femtosecond pulses developed a trailing "diffusive" exponential tail and the sum of (zeroth order) transmittance and reflectance was well below unity. These experimental findings have previously been ascribed to sample imperfections. Here, we analyze these findings by using 3D periodic approximants of the ideal photonic quasicrystals. We show that the experimental observations can be explained in terms of multiple scattering of light within these structures, i.e., in terms of intrinsic rather than purely extrinsic quasicrystal properties. PMID- 19189015 TI - Ronchigram quantification via a non-complementary dark-space effect. AB - We observe a non-complementary dark-space produced when two Ronchi-grams, at zero phase and pi-phase, are overlapped and use these dark spaces to quantify Ronchi grams. Diffraction and multiple beam interference effects narrow the Ronchi fringes created with a coherent point source illumination and prevent accurate determination of the geometrical fringe edges. The dark spaces created when the intensity of two Ronchi grams is added allow assessing the geometrical edge at the dark space middle providing a way to reduce measurement errors. We re construct the wavefront deformation in a test beam with a 35-term Zernike polynomial. PMID- 19189016 TI - Tailoring optical transmission via the arrangement of compound subwavelength hole arrays. AB - The transmission properties of light through metal films with compound periodic subwavelength hole arrays is numerically investigated by using the finite difference time-domain (FDTD) method. The sharp dips in the transmission bands, together with the suppression of surface Plasmon resonance (SPR) (0, 1) peak, are found when two square holes in every unit cell are arranged asymmetrically along the polarization direction of the incident light. However, the shape of transmission spectra is not sensitive to the symmetry if the holes are arranged perpendicular to the propagation direction of surface plasmon polaritons (SPPs). The physics origin of these phenomena is explained qualitatively by the phase resonance of SPPs. PMID- 19189017 TI - Efficient self-phase modulation in low loss, high index doped silica glass integrated waveguides. AB - We demonstrate efficient self phase modulation, as well as negligible nonlinear absorption, in low loss (<0.06 dB/cm), high index silica glass-based waveguides. Using approximately 1 ps pulses near 1560 nm we achieve a 1.5pi nonlinear phase shift in an integrated 45 cm long spiral waveguide with <60 W of peak input power, corresponding to a large nonlinearity (gamma) of 220 W(-1)km(-1). Further, we observe negligible nonlinear absorption for input intensities > 25 GW/cm(2). The high nonlinearity and low linear and nonlinear losses of these waveguides make them promising for nonlinear all-optical signal processing applications. PMID- 19189018 TI - Highly efficient optical-to-terahertz conversion in a sandwich structure with LiNbO3 core. AB - We demonstrate experimentally the efficiency of a recently proposed scheme of terahertz generation based on Cherenkov emission from ultrashort laser pulses in a sandwich structure. The structure has a thin nonlinear core covered with a prism of low terahertz absorption. Using an 8 mm long Si-LiNbO(3)-BK7 structure with a 50 microm thick LiNbO(3) core, we converted 40 microJ, 50 fs Ti:sapphire laser pulses into terahertz pulses of approximately 3 THz bandwidth with a record efficiency of over 0.1%. PMID- 19189019 TI - Astrophotonics: a new era for astronomical instruments. AB - Astrophotonics lies at the interface of astronomy and photonics. This burgeoning field has emerged over the past decade in response to the increasing demands of astronomical instrumentation. Early successes include: (i) planar waveguides to combine signals from widely spaced telescopes in stellar interferometry; (ii) frequency combs for ultra-high precision spectroscopy to detect planets around nearby stars; (iii) ultra-broadband fibre Bragg gratings to suppress unwanted background; (iv) photonic lanterns that allow single-mode behaviour within a multimode fibre; (v) planar waveguides to miniaturize astronomical spectrographs; (vi) large mode area fibres to generate artificial stars in the upper atmosphere for adaptive optics correction; (vii) liquid crystal polymers in optical vortex coronographs and adaptive optics systems. Astrophotonics, a field that has already created new photonic capabilities, is now extending its reach down to the Rayleigh scattering limit at ultraviolet wavelengths, and out to mid infrared wavelengths beyond 2500 nm. PMID- 19189020 TI - Sampling of the telescope image plane using single- and few-mode fibre arrays. AB - The coupling efficiency of starlight into single and few-mode fibres fed with lenslet arrays to provide a continuous field of view is investigated. The single mode field of view (FOV) and overall transmission is a highly complicated function of wavelength and fibre size leading to a continuous sample only in cases of poor throughput. Significant improvements are found in the few-mode regime with a continuous and efficient sample of the image plane shown to be possible with as few as 4 modes. This work is of direct relevance to the coupling of celestial light into photonic instrumentation and the removal of image scrambling and reduction of focal ratio degradation (FRD) using multi-mode fibre to single-mode fibre array converters. PMID- 19189021 TI - Optical Vectorial Vortex Coronagraphs using Liquid Crystal Polymers: theory, manufacturing and laboratory demonstration. AB - In this paper, after briefly reviewing the theory of vectorial vortices, we describe our technological approach to generating the necessary phase helix, and report results obtained with the first optical vectorial vortex coronagraph (OVVC) in the laboratory. To implement the geometrical phase ramp, we make use of Liquid Crystal Polymers (LCP), which we believe to be the most efficient technological path to quickly synthesize optical vectorial vortices of virtually any topological charge. With the first prototype device of topological charge 2, a maximum peak-to-peak attenuation of 1.4x10(-2) and a residual light level of 3x10(-5) at an angular separation of 3.5 lambda/d (at which point our current noise floor is reached) have been obtained at a wavelength of 1.55 microm. These results demonstrate the validity of using space-variant birefringence distributions to generate a new family of coronagraphs usable in natural unpolarized light, opening a path to high performance coronagraphs that are achromatic and have low-sensitivity to low-order wavefront aberrations. PMID- 19189022 TI - Octave-spanning supercontinuum generated in SF6-glass PCF by a 1060 nm mode locked fibre laser delivering 20 pJ per pulse. AB - We report the generation of an octave-spanning supercontinuum in SF6-glass photonic crystal fiber using a diode-pumped passively mode-locked fs Yb-fiber laser oscillating at 1060 nm. The pulses (energy up to 500 pJ and duration 60 fs) were launched into a 4 cm length of PCF (core diameter 1.7 microm and zero dispersion wavelength approximately 1060 nm). Less than 20 pJ of launched pulse energy was sufficient to generate a supercontinuum from 600 nm to 1450 nm, which represents the lowest energy so far reported for generation of an octave-spanning supercontinuum from a 1 microm pump. Since the laser pulse energy scales inversely with the repetition rate, highly compact and efficient sources based on SF6-glass PCF are likely to be especially useful for efficient spectral broadening at high repetition rates (several GHz), such as those needed for the precise calibration of astronomical spectrographs, where a frequency comb spacing >10 GHz is required for the best performance. PMID- 19189023 TI - Pupil remapping for high contrast astronomy: results from an optical testbed. AB - The direct imaging and characterization of Earth-like planets is among the most sought-after prizes in contemporary astrophysics, however current optical instrumentation delivers insufficient dynamic range to overcome the vast contrast differential between the planet and its host star. New opportunities are offered by coherent single mode fibers, whose technological development has been motivated by the needs of the telecom industry in the near infrared. This paper presents a new vision for an instrument using coherent waveguides to remap the pupil geometry of the telescope. It would (i) inject the full pupil of the telescope into an array of single mode fibers, (ii) rearrange the pupil so fringes can be accurately measured, and (iii) permit image reconstruction so that atmospheric blurring can be totally removed. Here we present a laboratory experiment whose goal was to validate the theoretical concepts underpinning our proposed method. We successfully confirmed that we can retrieve the image of a simulated astrophysical object (in this case a binary star) though a pupil remapping instrument using single mode fibers. PMID- 19189024 TI - Improved method for estimating the minimum length of modal filters fabricated for stellar interferometry. AB - We present an improved theoretical model to estimate the minimum fiber length required for achieving a desired degree of wavefront filtering in stellar interferometry. The proposed model is based on modal analysis of the fiber and is compared with numerical results obtained through the beam propagation method as well as with reported experimental observations. We also study the effect of introducing a spatial filter at the output end of the fiber and show that the required fiber length can be reduced significantly by introducing a circular aperture of optimum radius after the fiber. PMID- 19189025 TI - Mid-infrared guided optics: a perspective for astronomical instruments. AB - Research activities during the last decade have shown the strong potential of photonic devices to greatly simplify ground based and space borne astronomical instruments and to improve their performance. We focus specifically on the mid infrared wavelength regime (about 5-20 microm), a spectral range offering access to warm objects (about 300 K) and to spectral features that can be interpreted as signatures for biological activity (e.g. water, ozone, carbon dioxide). We review the relevant research activities aiming at the development of single-mode guided optics and the corresponding manufacturing technologies. We evaluate the experimentally achieved performance and compare it with the performance requirements for applications in various fields of astronomy. Our goal is to show a perspective for future astronomical instruments based on mid-infrared photonic devices. PMID- 19189026 TI - Ultrafast laser inscription: an enabling technology for astrophotonics. AB - The application of photonics to astronomy offers major advantages in the area of highly-multiplexed spectroscopy, especially when applied to extremely large telescopes. These include the suppression of the near-infrared night-sky spectrum [J. Bland-Hawthorn et al, Opt. Express 12, 5902 (2004), S. G. Leon-Saval et al, Opt. Lett. 30, 2545 (2005)] and the miniaturisation of spectrographs so that they may integrated into the lightpath of individual spatial samples [J. Bland Hawthorn et al, Proc SPIE 6269, 62690N (2006)]. Efficient collection of light from the telescope requires multimode optical fibres and three-dimensional photonic devices. We propose ultrafast laser inscription (ULI) [R. R. Thomson et al, Opt. Express 15, 11691 (2007)] as the best technology to fabricate 3D photonic devices for astrophotonic applications. PMID- 19189027 TI - Hybrid sol-gel planar optics for astronomy. AB - Hybrid sol-gel planar optics devices for astronomy are produced for the first time. This material system can operate from the visible (0.5 microm) up to the edge of astronomical J-band (1.4 microm). The design, fabrication and characterization results of a coaxial three beam combiner are given as an example. Fringe contrasts above 94% are obtained with a source with spectral bandwidth of 50 nm. These results demonstrate that hybrid sol-gel technology can produce devices with high quality, opening the possibility of rapid prototyping of new designs and concepts for astronomical applications. PMID- 19189028 TI - On-chip spectro-detection for fully integrated coherent beam combiners. AB - This paper presents how photonics associated with new arising detection technologies is able to provide fully integrated instrument for coherent beam combination applied to astrophysical interferometry. The feasibility and operation of on-chip coherent beam combiners has been already demonstrated using various interferometric combination schemes. More recently we proposed a new detection principle aimed at directly sampling and extracting the spectral information of an input signal together with its flux level measurement. The so called SWIFTS demonstrated concept that stands for Stationary-Wave Integrated Fourier Transform Spectrometer, provides full spectral and spatial information recorded simultaneously thanks to a motionless detecting device. Due to some newly available detection principles considered for the implementation of the SWIFTS concept, some technologies can even provide photo-counting operation that brought a significant extension of the interferometry domain of investigation in astrophysics. The proposed concept is applicable to most of the interferometric instrumental modes including fringe tracking, fast and sensitive detection, Fourier spectral reconstruction and also to manage a large number of incoming beams. The paper presents three practical implementations, two dealing with pair wise integrated optics beam combinations and the third one with an all-in-one 8 beam combination. In all cases the principles turned into a pair wise baseline coding after proper data processing. PMID- 19189029 TI - Efficient multi-mode to single-mode coupling in a photonic lantern. AB - We demonstrate the fabrication of a high performance multi-mode (MM) to single mode (SM) splitter or "photonic lantern", first described by Leon-Saval et al. (2005). Our photonic lantern is a solid all-glass version, and we show experimentally that this device can be used to achieve efficient and reversible coupling between a MM fiber and a number of SM fibers, when perfectly matched launch conditions into the MM fiber are ensured. The fabricated photonic lantern has a coupling loss for a MM to SM tapered transition of only 0.32 dB which proves the feasibility of the technology. PMID- 19189030 TI - Comparison of inverse scattering algorithms for designing ultrabroadband fibre Bragg gratings. AB - We compare different inverse scattering (IS) algorithms used to calculate profiles of fibre Bragg gratings (FBGs) and analyse their robustness, speed and implementation difficulties. We analyse sources of IS algorithm errors and discuss their relative importance. We discuss the optimal choice of IS algorithm for inverse-direct iterative optimisation schemes for grating design. We find that our time-domain layer-peeling method is an order of magnitude faster and more robust than the spectral domain algorithms considered here. We demonstrate that our method is essential to solving highly complex FBG designs demanded by astronomical applications. PMID- 19189031 TI - A new method for determining the transfer function of an acousto optical tunable filter. AB - The current study describes the determination of the transfer function of an Acousto Optical Tunable Filter from the in-flight solar observations of the SOIR instrument on board Venus Express. An approach is proposed in order to reconstruct the transfer function profile from the analysis of various solar lines. Moreover this technique allows the determination of the evolution of the transfer function as a function of the AOTF radio frequency. PMID- 19189032 TI - Sensitivity enhancement of nanoplasmonic sensors in low refractive index substrates. AB - Metal films perforated by nanoholes constitute a powerful platform for surface plasmon resonance biosensing. We find that the refractive index sensitivity of nanohole arrays increases if their resonance is red-shifted by increasing the separation distance between holes. However, an additional sensitivity enhancement occurs if the nanohole sensors are manufactured on low index substrates, despite the fact such substrates significantly blue-shift the resonance. We find a approximately 40% higher bulk refractive index sensitivity for a system of approximately 100 nm holes in 20 nm gold films fabricated on Teflon substrates (n=1.32) compared to the case when conventional glass substrates (n=1.52) are used. A similar improvement is observed for the case when a thin layer of dielectric material is deposited on the samples. These results can be understood by considering the electric field distribution induced by the so-called antisymmetric surface plasmon polariton in the thin gold films. PMID- 19189033 TI - Numerical simulation of an optical chromatographic separator. AB - Optical chromatography achieves microscale optical manipulation through the balance of optical and hydrodynamic forces on micron sized particles entrained in microfluidic flow traveling counter to the propagation of a mildly focused laser beam. The optical pressure force on a particle is specific to each particle's size, shape and refractive index. So far, these properties have been exploited in our lab to concentrate, purify and separate injected samples. But as this method advances into more complex optofluidic systems, a need to better predict behavior is necessary. Here, we present the development and experimental verification of a robust technique to simulate particle trajectories in our optical chromatographic device. We also show how this new tool can be used to gather better qualitative and quantitative understanding in a two component particle separation. PMID- 19189034 TI - Mutual optical injection in coupled DBR laser pairs. AB - We report an experimental study of nonlinear effects, characteristic of mutual optical coupling, in an ultra-short coupling regime observed in a distributed Bragg reflector laser pair fabricated on the same chip. Optical feedback is amplified via a double pass through a common on-chip optical amplifier, which introduces further nonlinear phenomena. Optical coupling has been introduced via back reflection from a cleave-ended fibre. The coupling may be varied in strength by varying the distance of the fibre from the output of the chip, without significantly affecting the coupling time. PMID- 19189035 TI - Thermo-recurrent nematic random laser. AB - This experimental work is aimed to investigate the thermal behavior of random laser action in dye doped nematic liquid crystals. The study evidenced an important temperature dependence of the random lasing characteristics in the nematic phase and in close proximity of the nematicisotropic (N-I) phase transition. A lowering of the laser emission intensity as the temperature increases is strictly related to the shift of the lasing threshold as function of the temperature even though the pump energy is kept fixed. The optical losses increasing owing to the thermal fluctuation enhanced scattering drive the input output smoother behavior until the system stops to lase, because below threshold. The unexpected reoccurrence of random lasing at higher temperature, in proximity of N-I transition is found to be related to a different scattering mechanism, the micro-droplets nucleation and critical opalescence. PMID- 19189036 TI - Responsiveness of the electronic Personal Assessment Questionnaire-Pelvic Floor (ePAQ-PF). AB - This study aimed to evaluate the responsiveness of the electronic Personal Assessment Questionnaire-Pelvic Floor (ePAQ-PF) in women undergoing surgery for pelvic floor disorders. Fifty-four women undergoing tension-free vaginal tape (TVT) and 47 women undergoing prolapse surgery completed ePAQ-PF at baseline and 3 months postoperatively. Responsiveness was calculated using effect sizes, standardized response means, responsiveness statistic, and minimally important difference. In the TVT group, the largest effect sizes were seen in the urinary domains for stress urinary incontinence (2.4), quality of life (2.2), and overactive bladder (0.9). In the prolapse group, the largest effect sizes were seen in the vaginal domains for prolapse (2.1) and quality of life (1.0). ePAQ-PF is responsive to change in women undergoing TVT or prolapse surgery in the salient and expected domains of stress incontinence and prolapse and quality of life and can be recommended for outcome measurement in this context. PMID- 19189037 TI - Treatment of osteoporosis with parathyroid hormone and teriparatide. AB - Nowadays osteoporosis treatment is based primarily on therapy with antiresorptive agents, like the bisphosphonates. Parathyroid hormone (Preotact) and human recombinant parathyroid hormone peptide 1-34 (Teriparatide) are relatively new for the treatment of osteoporosis and belong to the group of anabolic agents. Both agents demonstrated an increase in bone mineral density and a significant reduction in vertebral fractures in postmenopausal women with osteoporosis when given for 18-24 months. Data on nonvertebral fractures are, however, not clear cut, and so far only bisphosphonates and strontium ranelate have been demonstrated to reduce all types of fractures and therefore remain the front-line option for treatment of osteoporosis. As the safety, tolerability, and cost of the therapy also influence the choice of therapy, Preotact and Teriparatide might be useful additions to the armamentarium for (second-line) treatment of osteoporosis. PMID- 19189038 TI - Kinetics of calcium oxalate crystal growth in the presence of osteopontin isoforms: an analysis by scanning confocal interference microcopy. AB - Proteins that inhibit the growth and aggregation of calcium oxalate crystals play important roles in the prevention of kidney stone disease. One such protein is osteopontin (OPN), which inhibits the formation of calcium oxalate monohydrate (COM) in a phosphorylation-dependent manner. To determine the role of phosphate groups in the inhibition of COM growth by OPN, we used scanning confocal interference microscopy to compare the effects of highly phosphorylated OPN from cow milk, less phosphorylated OPN from rat bone, and nonphosphorylated recombinant OPN. COM growth was measured in the principal crystallographic directions <001>, <010>, and <100>, representing lattice-ion addition to {121}, {010}, and {100} faces, respectively. While the shapes of growth curves were very consistent from crystal to crystal, absolute growth rates varied widely. To control for this, results were expressed as changes in the aspect ratios <010>/<001> and <100>/<001>. Compared to control, bone OPN increased <010>/<001> and had no effect on <100>/<001>; milk OPN had no effect on <010>/<001>and decreased <100>/<001>; recombinant OPN had no significant effect on either aspect ratio. These findings indicate that milk OPN interacts with COM crystal faces in order of preference {100} > {121} approximately {010}, whereas bone OPN interacts in order of preference {100} approximately {121} > {010}. As {100} is the most Ca(2+)-rich face of COM, while {010} is the least Ca(2+)-rich, it appears that the OPN-mediated inhibition of COM growth occurs through a nonspecific electrostatic interaction between Ca(2+) ions of the crystal and phosphate groups of the protein. PMID- 19189039 TI - Lines of evidence for horizontal gene transfer of a phenazine producing operon into multiple bacterial species. AB - Phenazines are secondary metabolites with broad-spectrum antibiotic activity against bacteria, fungi, and eukaryotes. In pseudomonad species, a conserved seven-gene phenazine operon (phzABCDEFG) is required for the conversion of chorismic acid to the broad-spectrum antibiotic phenazine-1-carboxylate. Previous analyses of genes involved in phenazine production from nonpseudomonad species uncovered a high degree of sequence similarity to pseudomonad homologues. The analyses undertaken in this study wished to eluciadate the evolutionary history of genes involved in the production of phenazines. Furthermore, I wanted to determine if the phenazine operon has been transferred through horizontal gene transfer. Analyses of GC content, codon usage patterns, frequency of 3:1 dinucleotides, sequence similarities, and phylogenetic reconstructions were undertaken to map the evolutionary history of phenazine genes from multiple bacterial species. Patchy phyletic distribution, high sequence similarities, and phylogenetic evidence infer that pseudomonad, Streptomyces cinnamonensis, Pantoea agglomerans, Burkholderia cepacia, Pectobacterium atrosepticum, Brevibacterium linens, and Mycobacterium abscessus species all contain a phenazine operon which has most likely been transferred among these species through horizontal gene transfer. The acquisition of an antibiotic-associated operon is significant, as it may increase the relative fitness of the recipient species. PMID- 19189040 TI - Do polymorphisms in transcription factors LMX1A and LMX1B influence the risk for Parkinson's disease? AB - The key symptoms of Parkinson's disease (PD) are caused by degeneration of dopamine neurons originating in substantia nigra. Whereas, transcription factor LMX1A is crucial for the differentiation of mesencephalic dopamine neurons, LMX1B appears to be important for both the development and the survival of these cells. The aim of this study was to investigate if genetic variation in LMX1A and LMX1B differs between patients with PD (n = 357) and control subjects (n = 1428) by genotyping 33 single nucleotide polymorphisms (SNPs) in LMX1A and 11 SNPs in LMX1B. Three SNPs in LMX1A and one in LMX1B were associated with PD. After splitting for gender, six SNPs were associated with PD in women and four in men. The significances obtained did not survive correction for multiple testing, and our results should hence be interpreted with caution, but are partly in line with a previous report, and should thus be of sufficient interest to encourage further studies of these genes in PD. PMID- 19189041 TI - Does exposure to extremely low frequency magnetic fields produce functional changes in human brain? AB - Behavioral and neurophysiological changes have been reported after exposure to extremely low frequency magnetic fields (ELF-MF) both in animals and in humans. The physiological bases of these effects are still poorly understood. In vitro studies analyzed the effect of ELF-MF applied in pulsed mode (PEMFs) on neuronal cultures showing an increase in excitatory neurotransmission. Using transcranial brain stimulation, we studied noninvasively the effect of PEMFs on several measures of cortical excitability in 22 healthy volunteers, in 14 of the subjects we also evaluated the effects of sham field exposure. After 45 min of PEMF exposure, intracortical facilitation produced by paired pulse brain stimulation was significantly enhanced with an increase of about 20%, while other parameters of cortical excitability remained unchanged. Sham field exposure produced no effects. The increase in paired-pulse facilitation, a physiological parameter related to cortical glutamatergic activity, suggests that PEMFs exposure may produce an enhancement in cortical excitatory neurotransmission. This study suggests that PEMFs may produce functional changes in human brain. PMID- 19189042 TI - The sequential magnetic resonance images of tri-methyl tin leukoencephalopathy. AB - Organotin compounds are commonly used in industrial and agriculture. It causes toxic effects on skin, eyes, respiratory system, gastrointestinal system, and nervous system. After cleaning a di-methyl tin tank, 43-year-old man showed a dizziness, disorientation, visual hallucination, and agitation. Through a measurement by liquid chromatography and inductively coupled plasma-mass spectrometry, di-methyl tin and tri-methyl tin was detected. Although magnetic resonance (MR) image 3 days after exposure showed no abnormal signal intensity, follow-up MR images 15 days after exposure revealed abnormal extensive signal intensities in the white matter that was not ever coincident with previous reports. It was hardly explainable that previous abnormal signal intensities of MR image nearly disappeared 4 months later. We present a case of a patient who developed acute toxic leukoencephalopathy from an acute inhalational exposure to methyl tin with sequential MR images showing an involvement of white matter that was not ever reported. PMID- 19189043 TI - Levofloxacin-induced delirium. AB - An 83-year-old man was admitted for right lower lobe pneumonia which did not improve after a 5-day outpatient treatment with amoxicillin/clavulinate and clarithromycin. An empiric treatment with levofloxacin was started with a significant improvement after 24 h of this treatment. On the third day of hospitalization, delirium developed, while the patient was afebrile and with normal blood oxygenation. Treatment with levofloxacin was stopped, and a complete resolution of the patient's delirium was observed 2 days later. To the best of our knowledge, this is the third case of levofloxacin-induced delirium described in the medical literature. PMID- 19189044 TI - Predictors of caregiver burden in partners of patients with Parkinson's disease. AB - Aim of this study was to determine the predictors of caregiver burden among spouse caregivers of patients with Parkinson's disease (PD). Forty consecutive PD patients and their spouse caregivers were included. Patients were assessed using the Unified Parkinson's Disease Rating Scale (UPDRS), the Hoehn and Yahr scale (HY), the Mini-Mental State Examination (MMSE), the Neuropsychiatric Inventory (NPI), and the Geriatric Depression Scale (GDS). Stress and depressive symptoms among caregivers were evaluated using the Caregiver Burden Inventory (CBI) scale and the GDS. Only PD severity (HY) and mental symptoms (NPI) were significantly associated to caregiver distress. A major attention must be given to the early identification of factors generating stress in caregivers in order to improve caregiver quality of life and patient's care. PMID- 19189045 TI - Safety of carotid stenting for stroke prevention: need of an independent outcome assessor. AB - Safety and efficacy of carotid artery stenting have still to be fully established. We propose a standardized registry of carotid artery stenting in use at our hospital to evaluate whether the presence of an independent neurologist performing basal, procedural and post-procedural observation increases the accuracy of outcome assessment. We collected a cohort of patients receiving carotid stenting. An external neurologist supervised the endovascular intervention and monitored the patient's clinical conditions during procedure and follow-up time (12 months). The procedure was carried out successfully in all cases. We registered two intra-procedural strokes and two strokes within 24 h. The risk of major complications in our study was 9.1% at 30 days. Our complication rate is higher than in previous studies. These findings could be partly explained by the unemployment of distal protection devices, but also by the presence of an independent observer that might have increased the accuracy of neurological evaluation. PMID- 19189047 TI - Na+ mechanism of delta-opioid receptor induced protection from anoxic K+ leakage in the cortex. AB - Activation of delta-opioid receptors (DOR) attenuates anoxic K(+) leakage and protects cortical neurons from anoxic insults by inhibiting Na(+) influx. It is unknown, however, which pathway(s) that mediates the Na(+) influx is the target of DOR signal. In the present work, we found that, in the cortex, (1) DOR protection was largely dependent on the inhibition of anoxic Na(+) influxes mediated by voltage-gated Na(+) channels; (2) DOR activation inhibited Na(+) influx mediated by ionotropic glutamate N-methyl-D-aspartate (NMDA) receptors, but not that by non-NMDA receptors, although both played a role in anoxic K(+) derangement; and (3) DOR activation had little effect on Na(+)/Ca(2+) exchanger based response to anoxia. We conclude that DOR activation attenuates anoxic K(+) derangement by restricting Na(+) influx mediated by Na(+) channels and NMDA receptors, and that non-NMDA receptors and Na(+)/Ca(2+) exchangers, although involved in anoxic K(+) derangement in certain degrees, are less likely the targets of DOR signal. PMID- 19189048 TI - Tauroursodeoxycholic acid prevents E22Q Alzheimer's Abeta toxicity in human cerebral endothelial cells. AB - The vasculotropic E22Q mutant of the amyloid-beta (Abeta) peptide is associated with hereditary cerebral hemorrhage with amyloidosis Dutch type. The cellular mechanism(s) of toxicity and nature of the AbetaE22Q toxic assemblies are not completely understood. Comparative assessment of structural parameters and cell death mechanisms elicited in primary human cerebral endothelial cells by AbetaE22Q and wild-type Abeta revealed that only AbetaE22Q triggered the Bax mitochondrial pathway of apoptosis. AbetaE22Q neither matched the fast oligomerization kinetics of Abeta42 nor reached its predominant beta-sheet structure, achieving a modest degree of oligomerization with a secondary structure that remained a mixture of beta and random conformations. The endogenous molecule tauroursodeoxycholic acid (TUDCA) was a strong modulator of AbetaE22Q-triggered apoptosis but did not significantly change the secondary structures and fibrillogenic propensities of Abeta peptides. These data dissociate the pro-apoptotic properties of Abeta peptides from their distinct mechanisms of aggregation/fibrillization in vitro, providing new perspectives for modulation of amyloid toxicity. PMID- 19189046 TI - Role of protein-tyrosine phosphatases in regulation of osteoclastic activity. AB - Osteoclasts, the primary cell type mediating bone resorption, are multinucleated, giant cells derived from hematopoietic cells of monocyte-macrophage lineage. Osteoclast activity is, in a large part, regulated by protein-tyrosine phosphorylation. While information about functional roles of several protein tyrosine kinases (PTK), including c-Src, in osteoclastic resorption has been accumulated, little is known about the roles of protein-tyrosine phosphatases (PTPs) in regulation of osteoclast activity. Recent evidence implicates important regulatory roles for four PTPs (SHP-1, cyt-PTP-epsilon, PTP-PEST, and PTPoc) in osteoclasts. Cyt-PTP-epsilon, PTP-PEST, and PTP-oc are positive regulators of osteoclast activity, while SHP-1 is a negative regulator. Of these PTPs in osteoclasts, only PTP-oc is a positive regulator of c-Src PTK through dephosphorylation of the inhibitory phosphotyrosine-527 residue. Although some information about mechanisms of action of these PTPs to regulate osteoclast activity is reviewed in this article, much additional work is required to provide more comprehensive details about their functions in osteoclasts. PMID- 19189049 TI - The PAS-domain kinase PASKIN: a new sensor in energy homeostasis. AB - The PAS domain kinase PASKIN, also termed PAS kinase or PASK, is an evolutionarily conserved potential sensor kinase related to the heme-based oxygen sensors of nitrogen-fixing bacteria. In yeast, the two PASKIN homologs link energy flux and protein synthesis following specific stress conditions. In mammals, PASKIN may regulate glycogen synthesis and protein translation. Paskin knock-out mice do not show any phenotype under standard animal husbandry conditions. Interestingly, these mice seem to be protected from the symptoms of the metabolic syndrome when fed a high-fat diet. Energy turnover might be increased in specific PASKIN-deficient cell types under distinct environmental conditions. According to the current model, binding of a putative ligand to the PAS domain disinhibits the kinase domain and activates PASKIN auto- and target phosphorylation. Future research needs to be conducted to elucidate the nature of the putative ligand and the molecular mechanisms of downstream signalling by PASKIN. PMID- 19189050 TI - Interaction with PFK-/FBP-2 is essential to glucokinase molecular physiology. PMID- 19189051 TI - Fibulins: multiple roles in matrix structures and tissue functions. AB - The fibulins are a family of secreted glycoproteins associated with basement membranes, elastic fibers, and other matrices. They are expressed in a variety of tissues. Association with these matrix structures is mediated by their ability to interact with many extracellular matrix constituents. The seven members of the family are defined by the presence of two structural modules, a tandem repeat of epidermal growth factor-like modules and a unique C-terminal fibulin-type module. They act not only as intermolecular bridges within the extracellular matrix to form supramolecular structures, but also as mediators for cellular processes and tissue remodeling. These important functions of fibulins in a wide range of biological processes have been shown in in vitro systems, gene knockout mice, and human genetic disorders. In this review, we describe the structure and function of these proteins and discuss the implication of fibulins in development and diseases. PMID- 19189052 TI - Chronic granulomatous disease. AB - Chronic granulomatous disease is an inherited disorder of the NADPH oxidase characterized by severe bacterial and fungal infections and disordered inflammation. We propose that NADPH oxidase has a key role in regulating acute neutrophilic and T cell responses, which in turn restrains fungal growth and calibrates the inflammatory response to minimize injury and allergy. In this model, superoxide-induced activation of indoleamine 2,3-dioxygenase (IDO) is a central mechanism by which the optimal balance of antifungal host defense and immune tolerance occurs. This model is based on studies in mice and requires correlation in humans. PMID- 19189053 TI - Presenilin-dependent regulated intramembrane proteolysis and gamma-secretase activity. AB - Inhibiting the production of amyloid-beta by antagonising gamma-secretase activity is currently being pursued as a therapeutic strategy for Alzheimer's disease (AD). However, early pre-clinical studies have demonstrated that disruption of presenilin-dependent gamma-secretase alters many presenilin dependent processes, leading to early lethality in several AD model organisms. Subsequently, transgenic animal studies have highlighted several gross developmental side effects arising from presenilin deficiency. Partial knockdown or tissue-specific knockout of presenilins has identified the skin, vascular and immune systems as very sensitive to loss of presenilin functions. A more appreciative understanding of presenilin biology is therefore demanded if gamma secretase is to be pursued as a therapeutic target. Herein we review the current understanding of gamma-secretase complexes; their regulation, abundance of interacting partners and diversity of substrates. We also discuss regulation of the gamma-secretase complexes, with an emphasis on the functional role of presenilins in cell biology. PMID- 19189054 TI - A metabolically stable analogue of anandamide, Met-F-AEA, inhibits human thyroid carcinoma cell lines by activation of apoptosis. AB - The active components of Cannabis sativa and their derivatives produce a wide spectrum of effects, some of which may have clinical application. The discovery of specific cannabinoid receptors and a family of endogenous ligands of those receptors has attracted much attention to cannabinoids as agents capable of controlling the decision of cells to survive or die. We analysed the effects exerted by 2-methyl-2'-F-anandamide (Met-F-AEA), a metabolically stable analogue of anandamide, and observed a growth inhibition in cell lines derived from thyroid carcinomas. Growth inhibition was associated with a high level of CB1 receptor expression, suggesting that the cytotoxic effect is due to interaction with the CB1 receptor. This phenomenon was associated with activation of the protein, p53, an increased apoptotic rate, and expression of p21(CIP1/WAF1). This study provides new insights into the mechanism of Met-F-AEA action, and could have significance in providing a basis for the management of thyroid carcinoma. PMID- 19189055 TI - Initial experience with bevacizumab treatment for biopsy confirmed cerebral radiation necrosis. AB - BACKGROUND: Cerebral radiation necrosis is a serious complication of radiation treatment for brain tumors. Therapeutic options include corticosteroids, anticoagulation and hyperbaric oxygen with limited efficacy. Bevacizumab, an antibody against VEGF had been reported to reduce edema in patients with suspected radiation necrosis. We retrospectively reviewed 6 patients with biopsy proven cerebral radiation necrosis treated with bevacizumab between 2006 and 2008. RESULTS: Interval MRI follow-up demonstrated radiographic response in all patients with an average reduction of 79% for the post gadolinium studies and 49% for the FLAIR images. The initial partial radiographic response was noted for up to a mean follow-up time of 5.9 months (6 weeks to 18 months). CONCLUSION: Bevacizumab appears to produce radiographic response and clinical benefits in the treatment of patients with cerebral radionecrosis. PMID- 19189056 TI - Effect of lactoferrin on oxidative features of ceruloplasmin. AB - In our previous report we first described a complex between lactoferrin (Lf) and ceruloplasmin (Cp) with K (d) approximately 1.8 microM. The presence of this complex in colostrum that never contains more than 0.3 microM Cp questions the reliability of K (d) value. We carefully studied Lf binding to Cp and investigated the enzymatic activity of the latter in the presence of Lf, which allowed obtaining a new value for K (d) of Cp-Lf complex. Lf interacting with Cp changes its oxidizing activity with various substrates, such as Fe(2+), o dianisidine (o-DA), p-phenylenediamine (p-PD) and dihydroxyphenylalanine (DOPA). The presence of at least two binding sites for Lf in Cp molecule is deduced from comparison of substrates' oxidation kinetics with and without Lf. When Lf binds to the first site affinity of Cp to Fe(2+) and to o-DA increases, but it decreases towards DOPA and remains unchanged towards p-PD. Oxidation rate of Fe(2+) grows, while that of o-DA, p-PD and DOPA goes down. Subsequent Lf binding to the second center has no effect on iron oxidation, hampers DOPA and o-DA oxidation, and reduces affinity towards p-PD. Scatchard plot for Lf sorbing to Cp Sepharose allowed estimating K (d) for Lf binding to high-affinity (approximately 13.4 nM) and low-affinity (approximately 211 nM) sites. The observed effect of Lf on ferroxidase activity of Cp is likely to have physiological implications. PMID- 19189058 TI - A new aminopeptidase from the keratin-degrading strain Streptomyces fradiae var. k11. AB - An aminopeptidase gene fragment was isolated from a keratin-degrading strain, Streptomyces fradiae var. k11, by PCR amplification using a degenerate primer set designed based on the partial amino acid sequence of the native enzyme. The gene, designated sfap, encoded a polypeptide of 461 amino acids comprised of three domains: a signal peptide, a mature region, and a C-terminal propeptide. The aminopeptidase, SFAP, had highest amino acid sequence identity (79%) with a putative aminopeptidase from Streptomyces griseus subsp. griseus NBRC 13350. The gene with and without C-terminal propeptide was successfully overexpressed in Escherichia coli BL21 (DE3), and the gene without C-terminal propeptide encoded a functional enzyme. Purified recombinant SFAP exhibited optimal activity at pH 8.0 and 60 degrees C, and retained >60% peak activity over a broad range of temperature. The enzyme was thermal and pH stable, and showed metalloprotease characteristics, which was inhibited by EDTA but activated by Ca(2+) and Co(2+). This is the first study to report the gene cloning and expression of a leucine aminopeptidase from S. fradiae. PMID- 19189057 TI - Xenopus, a unique comparative model to explore the role of certain heat shock proteins and non-classical MHC class Ib gene products in immune surveillance. AB - The heat shock proteins (HSPs) gp96 and hsp70 can elicit potent anti-tumor responses and as such have significant clinical potential. Besides cytotoxic CD8 T cell (CTLs) effectors, evidence suggests that natural killer (NK) cells and other less well-characterized cell types also play a critical role in HSP mediated anti-tumor responses. Owing to their high degree of phylogenetic conservation, we have proposed that HSPs are ancestral agents of immune surveillance; and postulated that their immunological properties, if important, should have been conserved during evolution. We are investigating this issue using a unique non-mammalian comparative tumor-immunity model in the frog Xenopus, which allows us to focus on the relationship between HSPs, classical MHC class Ia, and non-classical MHC class Ib molecules. In addition to a transplantable lymphoid tumor in genetically defined cloned Xenopus, we are generating transgenic frogs with inducible or knocked-down (RNAi) gene expression. PMID- 19189059 TI - Improvement of steroid biotransformation with hydroxypropyl-beta-cyclodextrin induced complexation. AB - The inclusion complexes induced by cyclodextrins and its derivates have been shown previously to enhance the biotransformation of hydrophobic compounds. Using hydroxypropyl-beta-cyclodextrin (HP-beta-CD; 20% w/v), the water solubility of cortisone acetate increased from 0.039 to 7.382 g L(-1) at 32 degrees C. The solubilization effect of HP-beta-CD was far superior to dimethylformamide (DMF) and ethanol. The dissolution rate also significantly increased in the presence of HP-beta-CD. The enzymatic stability of Delta(1)-dehydrogenase from Arthrobacter simplex TCCC 11037 was not influenced by the increasing concentrations of HP-beta CD contrary to the organic cosolvents which negatively influenced in the order DMF > ethanol. The activity inhibition effect caused by HP-beta-CD was not so conspicuous as ethanol and DMF. Inactivation constants of ethanol, DMF, and HP beta-CD were 5.832, 4.541, and 1.216, respectively. The inactivation energy (E (a)) was in the order of HP-beta-CD (55.1 kJ mol(-1)) > ethanol (39.9 kJ mol(-1)) > DMF (37.1 kJ mol(-1)). PMID- 19189060 TI - Rapid screening and isolation of a fungus for sitosterol to androstenedione biotransformation. AB - Androstenedione (AD) is a steroid intermediate used in the pharmaceutical industry for the production of several important anabolic drugs. An important route for producing AD is by the microbial transformation of sterols. Microbes capable of efficiently transforming sterols to AD are few and newer strains need to be isolated. Conventional procedures for screening and isolation are time consuming. A new procedure was used for screening and isolation of fungal microorganisms capable of biotransformation of sitosterol to androstenedione. In this procedure, Basic Alignment Search Tool (BLAST) and Position-Specific Iterative BLAST were employed to obtain a parent set of candidate microorganisms. The parent set was reduced using heuristics and constraints to obtain a manageable number of microorganisms that may be tested experimentally. For this work, screening of the entire NCBI database yielded a parent set containing 64 microorganisms. Among these, only two microorganisms, Aspergillus oryzae and Aspergillus nidulans FGSC A4, qualified to the experimental stage. Sitosterol biotransformation experiments were carried out using A. oryzae and the production of AD in culture medium was confirmed. PMID- 19189061 TI - [Teaching non-technical skills for critical incidents: Crisis resource management training for medical students]. AB - BACKGROUND: Physicians have to demonstrate non-technical skills, such as communication and team leading skills, while coping with critical incidents. These skills are not taught during medical education. A crisis resource management (CRM) training was established for 4th to 6th year medical students using a full-scale simulator mannikin (Emergency Care Simulator, ECS, METI). PATIENTS AND METHODS: The learning objectives of the course were defined according to the key points of Gaba's CRM concept. The training consisted of theoretical and practical parts (3 simulation scenarios with debriefing). Students' self-assessment before and after the training provided the data for evaluation of the training outcome. RESULTS: A total of 65 students took part in the training. The course was well received in terms of overall course quality, debriefings and didactic presentation, the mean overall mark being 1.4 (1: best, 6: worst). After the course students felt significantly more confident when facing incidents in clinical practice. The main learning objectives were achieved. CONCLUSION: The effectiveness of applying the widely used ECS full scale simulator in interdisciplinary teaching has been demonstrated. The training exposes students to crisis resource management issues and motivates them to develop non-technical skills. PMID- 19189063 TI - [Damage of the subclavian vein with a thorax drainage]. AB - The necessity for insertion of a thorax drainage can lead to serious complications. Especially the use of a trocar instead of a mini-thoracotomy has a notable risk of misplacement or complications. Chest tubes per se can perforate the parietal pleura resulting in an extrathoracic position of the tube and contraindications for chest tubes should always be considered. Accidental lacerations of blood vessels should be avoided by a correct technique and a clear indication. A chest X-ray in one plane is not sufficient to prove correct positioning of a chest tube and suspected misplacement of a chest tube has to be assessed radiographically in at least two planes. A case of a fatal complication of a misplaced chest tube causing a laceration of the right subclavian vein is described. PMID- 19189062 TI - [Leipzig fast-track protocol for cardio-anesthesia. Effective, safe and economical]. AB - BACKGROUND: In November 2005 a complex, multimodal anesthesia fast-track protocol (FTP) was introduced for elective cardiac surgery patients in the Cardiac Center of the University of Leipzig which included changing from an opioid regime to remifentanil and postoperative treatment in a special post-anesthesia recovery and care unit. The goal was to speed up recovery times while maintaining safety and improving costs. METHOD: A total of 421 patients who underwent the FTP and were treated in the special recovery room were analyzed retrospectively. These patients were compared with patients who had been treated by a standard protocol (SP) prior to instituting the FTP. Primary outcomes were time to extubation, length of stay in the intensive care unit (ICU) and treatment costs. RESULTS: The times to extubation were significantly shorter in the FTP group with 75 min (range 45-110 min) compared to 900 min (range 600-1140 min) in the SP group. Intensive care unit stay and hospital length of stay were also significantly shorter in the FTP group (p<0.01). The reduction of treatment costs of intensive care for FTP patients was 53.5% corresponding to savings of EUR 738 per patient in the FTP group compared with the SP group. CONCLUSIONS: The Leipzig fast-track protocol for cardio-anesthesia including the central elements of switching opiate therapy to remifentanil and switching patient recovery to a special post anesthesia recovery and care unit, shortened therapy times, is safe and economically effective. PMID- 19189064 TI - [Intensive care medicine as a component of the compulsory medical curriculum. Evaluation of a pilot curriculum at the University Hospital Aachen]. AB - BACKGROUND: In order to provide early achievement of practical experience during medical education, the medical faculty of the university Aachen has developed a new medical school curriculum which was offered in 2003 for the first time. In this curriculum anaesthesiology became a compulsory subject with practical training both in the operation theatre and in emergency medicine. Accordingly, a practical course in the field of intensive care medicine has also been designed with respect to the planned schedule and personnel resources. This course was evaluated by both students and teaching staff in a written, anonymous form as a quality control. METHODS: A dedicated course was developed for medical students of the 8th and 9th semesters. In this course comprised of 6 students and lasting 1 week, practical training is provided by intensive care physicians and accompanied by theoretical lessons focusing on the definition, diagnosis, therapy and prophylaxis of sepsis, essentials of mechanical ventilation and patient presentation at the bedside during daily rounds. On the last day of training students were required to present patients by themselves thereby recapitulating the acquired knowledge. In the summer semester 2007 this intensive care training course was offered for the first time. All participating 83 students and 23 physicians involved in teaching evaluated the course with marks from 1 to 6 according to the standard German school grading system using an online questionnaire. RESULTS: Students rated the course with 1.6+/-0.7 (mean +/- SD) for comprehensibility, with 1.6+/-0.7 for structural design, and with 1.7+/-0.7 for agreement between teachers. They graded their personal learning success with 1.7+/-0.7. With a cumulative mark of 1.7+/-0.6, the course was ranked as 1 of the top 3 courses of the medical faculty from the very beginning. The majority of the teaching staff (80%) appreciated the focus on few selected teaching subjects. However, comprehensibility, structural design, agreement between teachers and personal learning success were graded one mark worse than by the students. CONCLUSIONS: According to the results, efficiency and acceptance of intensive care training courses were high. Major criteria for the high grading were a limited number of participants, the focus on few subjects, and a clear structural design. However, according to several personal notes from the students, simulation-based sessions and written teaching material might further improve success of this course. PMID- 19189065 TI - [Propofol administration systems. Handling, hemodynamics and propofol consumption]. AB - BACKGROUND: During anaesthesia propofol is administered either by manual controlled infusion (MCI) or by target controlled infusion (TCI) techniques. In this study two different TCI systems for propofol administration were evaluated with regard to handling, patient safety, and costs and compared to administration of propofol by the MCI technique. METHODS: In a prospective study, 90 patients scheduled for elective surgery of the nose or nasal sinuses were randomly enrolled in three groups. The two TCI systems were examined in two groups of 30 patients: one group received propofol following the pharmacokinetic TCI model of Schnider (TCI-Schnider) and the other group received propofol following the TCI model of Marsh (TCI-Marsh). A manual perfusion technique (MCI, n=30) was used in the control group. Depth of anesthesia was controlled using the bispectral index (BSI) which was adjusted to fall within the range of 40-55. Hemodynamics, extubation times and time of awaking, rate and quality of propofol dose adjustment, total drug requirements, costs, and quality of recovery were documented. The incidence of postoperative nausea and vomiting (PONV) as well as shivering and patient satisfaction were also documented. RESULTS: Demographics, hemodynamics and perioperative data did not differ between the groups. Propofol consumption within the first 60 min also showed no significant differences. In the course of extended anaesthesia, propofol consumption was significantly less in both TCI groups compared to the control group (MCI) and the TCI-Schnider group also showed less episodes of bradycardia. The necessity of propofol dose adjustment did not differ significantly between the TCI groups. Administration and consumption of anaesthesia co-medication (fentanyl, remifentanil, cisatracurium) did not differ between the groups. CONCLUSION: The investigated propofol administration procedures using the MCI or TCI techniques were safe and easy to handle under BIS monitoring. No differences were found concerning extubation times and time of awaking. During extended anaesthesia procedures (>60 min), propofol consumption was lower with both TCI techniques and thus costs could be saved. PMID- 19189066 TI - [Severe sepsis and disseminated intravascular coagulation. Supplementation with antithrombin]. AB - Administration of high-dose antithrombin (AT) was investigated on a large collective of patients with severe sepsis in the KyberSept study. In the total study the administration of AT resulted in no significant reduction of the mortality rate in comparison to a placebo. However, in the protocol of this study subgroups were predefined, which when analyzed revealed that the group of patients who received AT but not simultaneously heparin did show a reduction of the mortality rate in comparison to the placebo group. The reduction of the absolute mortality rate of 15% reached statistical significance on day 90. Even patients classified as risk group grade II according to the Simplified Acute Physiology Score (SAPS), showed a significant reduction of the mortality rate of approximately 22% after 90 days without simultaneous administration of heparin. Such a positive result for administration of AT without simultaneous heparin treatment can also be found when severe sepsis complicated by disseminated intravascular coagulation (DIC) is present. Coagulation diagnostic assists the recognition of latent or fulminant DIC and also in surveillance of the course and development. The results of AT supplementation for severe sepsis and DIC are in agreement with earlier studies on smaller patient collectives and suggest that a randomized controlled clinical study should be carried out on a subcollective of severely ill patients. PMID- 19189067 TI - [In memoriam A. Bernard Ackerman: 1936-2008]. PMID- 19189068 TI - [Scabies]. AB - Scabies is an infectious disease caused by Sarcoptes scabiei var. hominis. The disease is mainly transmitted by close personal contact. The primary clinical picture is characterized by burrows at typical areas as well as severe pruritus, while secondarily a rather non-specific exanthema can be observed. The detection of mites or mite products (eggs, feces) confirms the diagnosis; dermatoscopy may facilitate the diagnosis. First choice therapy for most variants of scabies is topical permethrin. Oral administration of ivermectin is recommended for some special indications. In the following article, the current knowledge about epidemiology, biology, immunology, clinical appearance, diagnostic procedures and therapy of scabies is reviewed. PMID- 19189069 TI - [Psoriasis in childhood and adolescence: clinical features and therapy]. AB - Psoriasis starts in the first two decades of life in about one-third of patients. Initially the diagnosis may be difficult because of atypical or mild involvement. Plaque psoriasis is the most frequent type in children, just as in adults. However, lesions are often smaller, thinner and less scaly. Treatment of childhood psoriasis represents a special challenge to dermatologists since many therapeutic agents are not approved and guidelines are generally lacking for this age group. Apart from clinical type, severity of the disease and patient's age, the choice of treatment must be based on safety concerns and accessibility of treatment. We herein focus on the peculiarities of clinical presentations and on the management of psoriasis in children and adolescents. PMID- 19189070 TI - NF-kappaB blockade upregulates Bax, TSP-1, and TSP-2 expression in rat granulation tissue. AB - Several diseases are characterized by chronic inflammation, a condition frequently associated with angiogenesis and fibrogenesis that account for the development of granulation tissue. Nuclear Factor kappa-light-chain-enhancer of activated B cells (NF-kappaB) is a crucial modulator of intracellular prosurvival signaling pathways and is implicated in the pathogenesis of inflammatory process. In this study, we have investigated the role of NF-kappaB in the angiogenic and fibrogenic response induced by lambda-carrageenin in a rat model of chronic inflammation at 1, 3, and 5 days. The subcutaneous implant of lambda-carrageenin soaked sponges in rat induced a time-related increase of granulation tissue formation accompanied by intense neovascularization. These lambda-carrageenin induced changes were significantly reduced by coinjection of wild-type oligodeoxynucleotide (WT ODN) decoy to NF-kappaB. Molecular, morphological, and ultrastructural analysis performed on whole granulation tissue demonstrated: (1) inhibition of NF-kappaB/DNA binding activity; (2) downregulation of cyclooxygenase-2, matrix metalloproteinase-9, tumor necrosis factor-alpha, and vascular endothelial growth factor; (3) upregulation of thrombospondin (TSP)-1 at 1 day and TSP-2 at 5 days; and (4) increase in Bax to Bcl-2 ratio. Our findings show that the blockade of NF-kappaB activation by WT ODN decoy prevents the development of granulation tissue induced by lambda-carrageenin-soaked sponge implant upregulating Bax as well as TSP-1 and TSP-2 expression. PMID- 19189071 TI - Functions of C5a receptors. AB - The split product of the complement protein, C5, is C5a and is an extremely potent pro-inflammatory peptide that interacts with two C5a receptors, C5aR and C5L2, present on surfaces of phagocytes as well as other cell types. The former is a well-established receptor that initiates G-protein-coupled signaling via mitogen-activated protein kinase pathways. Its in vivo blockade greatly reduces inflammatory injury. Much less is known about C5L2, occupancy of which by C5a does not initiate increased intracellular Ca(2+). There are numerous conflicting reports suggesting that C5L2 is a "default receptor" that attenuates C5a dependent biological responses by competing with C5aR for binding of C5a. However, there are other reports suggesting that C5L2 plays an active, positive role in inflammatory responses. Better definition of C5L2 is needed if its in vivo blockade, along with C5aR, is to be considered in complement-dependent inflammatory diseases. PMID- 19189072 TI - Tumor initiating cells in malignant gliomas: biology and implications for therapy. AB - A rare subpopulation of cells within malignant gliomas, which shares canonical properties with neural stem cells (NSCs), may be integral to glial tumor development and perpetuation. These cells, also known as tumor initiating cells (TICs), have the ability to self-renew, develop into any cell in the overall tumor population (multipotency), and proliferate. A defining property of TICs is their ability to initiate new tumors in immunocompromised mice with high efficiency. Mounting evidence suggests that TICs originate from the transformation of NSCs and their progenitors. New findings show that TICs may be more resistant to chemotherapy and radiation than the bulk of tumor cells, thereby permitting recurrent tumor formation and accounting for the failure of conventional therapies. The development of new therapeutic strategies selectively targeting TICs while sparing NSCs may provide for more effective treatment of malignant gliomas. PMID- 19189073 TI - Receptor for AGE (RAGE) and its ligands-cast into leading roles in diabetes and the inflammatory response. AB - The actors in the pathogenesis of diabetes and its complications are many and multifaceted. The effects of elevated levels of glucose are myriad; among these is the generation of advanced glycation end products (AGEs), the products of nonenzymatic glycoxidation of proteins and lipids. The finding that AGEs stimulate signal transduction cascades through the multiligand receptor RAGE unveiled novel insights into diabetes and its complications. Inextricably woven into AGE-RAGE interactions in diabetes is the engagement of the innate and adaptive immune responses. Although glucose may be the triggering stimulus to draw RAGE into diabetes pathology, consequent cellular stress results in release of proinflammatory RAGE ligands S100/calgranulins and HMGB1. We predict that once RAGE is engaged in the diabetic tissue, a vicious cycle of ligand-RAGE perturbation ensues, leading to chronic tissue injury and suppression of repair mechanisms. Targeting RAGE may be a beneficial strategy in diabetes, its complications, and untoward inflammatory responses. PMID- 19189074 TI - Decreased contractility due to energy deprivation in a transgenic rat model of hypertrophic cardiomyopathy. AB - Hypertrophic cardiomyopathy (HCM) is associated with cardiac hypertrophy, diastolic dysfunction, and sudden death. Recently, it has been suggested that inefficient energy utilization could be a common molecular pathway of HCM-related mutations. We have previously generated transgenic Sprague-Dawley rats overexpressing a truncated cardiac troponin T (DEL-TNT) molecule, displaying typical features of HCM such as diastolic dysfunction and an increased susceptibility to ventricular arrhythmias. We now studied these rats using 31P magnetic resonance spectroscopy (MRS). MRS demonstrated that cardiac energy metabolism was markedly impaired, as indicated by a decreased phosphocreatine to ATP ratio (-31%, p < 0.05). In addition, we assessed contractility of isolated cardiomyocytes. While DEL-TNT and control cardiomyocytes showed no difference under baseline conditions, DEL-TNT cardiomyocytes selectively exhibited a decrease in fractional shortening by 28% after 1 h in glucose-deprived medium (p < 0.05). Moreover, significant decreases in contraction velocity and relaxation velocity were observed. To identify the underlying molecular pathways, we performed transcriptional profiling using real-time PCR. DEL-TNT hearts exhibited induction of several genes critical for cardiac energy supply, including CD36, CPT-1/-2, and PGC-1alpha. Finally, DEL-TNT rats and controls were studied by radiotelemetry after being stressed by isoproterenol, revealing a significantly increased frequency of arrhythmias in transgenic animals. In summary, we demonstrate profound energetic alterations in DEL-TNT hearts, supporting the notion that inefficient cellular ATP utilization contributes to the pathogenesis of HCM. PMID- 19189075 TI - [Targeting B cells in multiple sclerosis. Current concepts and strategies]. AB - Multiple sclerosis (MS) is a chronic inflammatory demyelinating autoimmune disease of the CNS and a leading cause of lasting neurological disability in younger adults. In the last decade our knowledge of its immunopathogenesis expanded vastly. It is now widely appreciated that B cells are key players in the autoreactive immune network. They exert far more functions than merely being the precursors of antibody-producing plasma cells. B cells act as efficient antigen presenting cells and may stimulate an autoreactive immune response through secretion of proinflammatory cytokines. It is thus only logical to test therapeutic strategies targeting B cells in MS. Rituximab is a depleting chimeric monoclonal antibody directed against CD20 and expressed on developing, naive, and memory B cells but not stem or plasma cells. Several smaller studies have been conducted that led to a placebo controlled, double blind phase II study on efficacy which was reported recently. The results are very promising, meeting not only the primary endpoint of reduction of the surrogate MRI marker of contrast enhancing lesions but also showing a reduction in clinical relapse rate of patients treated with rituximab. This review discusses the role of autoreactive B cells in the context of MS, analyzes the B-cell-depleting treatment studies reported, and provides information on planned and future B-cell-directed therapeutic strategies in MS. PMID- 19189076 TI - [Hallucinosis using lamotrigine]. PMID- 19189077 TI - Arterial blood pressure during early sepsis and outcome. AB - OBJECTIVE: To evaluate the association between arterial blood pressure (ABP) during the first 24 h and mortality in sepsis. DESIGN: Retrospective cohort study. SETTING: Multidisciplinary intensive care unit (ICU). PATIENTS AND PARTICIPANTS: A total of 274 septic patients. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: Hemodynamic, and laboratory parameters were extracted from a PDMS database. The hourly time integral of ABP drops below clinically relevant systolic arterial pressure (SAP), mean arterial pressure (MAP), and mean perfusion pressure (MPP = MAP - central venous pressure) levels was calculated for the first 24 h after ICU admission and compared with 28-day-mortality. Binary and linear regression models (adjusted for SAPS II as a measure of disease severity), and a receiver operating characteristic (ROC) analysis were applied. The areas under the ROC curve were largest for the hourly time integrals of ABP drops below MAP 60 mmHg (0.779 vs. 0.764 for ABP drops below MAP 55 mmHg; P < or = 0.01) and MPP 45 mmHg. No association between the hourly time integrals of ABP drops below certain SAP levels and mortality was detected. One or more episodes of MAP < 60 mmHg increased the risk of death by 2.96 (CI 95%, 1.06-10.36, P = 0.04). The area under the ROC curve to predict the need for renal replacement therapy was highest for the hourly time integral of ABP drops below MAP 75 mmHg. CONCLUSIONS: A MAP level > or = 60 mmHg may be as safe as higher MAP levels during the first 24 h of ICU therapy in septic patients. A higher MAP may be required to maintain kidney function. PMID- 19189078 TI - The impact of obesity on outcomes after critical illness: a meta-analysis. AB - PURPOSE: To assess whether obesity is associated with mortality or other adverse intensive care unit (ICU) and post-ICU outcomes. METHODS: A meta-analysis of studies from PubMed and EMBASE databases. RESULTS: Twenty-two studies (n = 88,051 patients) were included. Pooled analysis demonstrated no difference in ICU mortality, but lower hospital mortality for obese and morbidly obese subjects (RR 0.76; 95% CI 0.59, 0.92; RR 0.83; 95% CI 0.66, 1.04, respectively) versus normal weight subjects. There was no association between obesity and duration of mechanical ventilation or ICU stay. Morbidly obese versus normal weight patients had longer hospitalizations. No study reported physical function, mental health, or quality of life outcomes after discharge. CONCLUSIONS: Obesity is not associated with increased risk for ICU mortality, but may be associated with lower hospital mortality. There is a critical lack of research on how obesity may affect complications of critical illness and patient long-term outcomes. PMID- 19189079 TI - Bacterial ventilator-associated pneumonia: bronchoalveolar lavage results are not influenced by dilution. AB - OBJECTIVE: This study was designed to determine if bronchoalveolar lavage (BAL) quantitative culture results can be used confidently for the diagnosis of bacterial ventilator-associated pneumonia (VAP) without taking dilution into account. DESIGN: Prospective observational cohort study. SETTING: A 12-bed medical ICU in a teaching hospital. PATIENTS: A total of 241 BAL (three 50-mL aliquots) were performed in 127 patients presenting a suspicion of VAP. INTERVENTIONS: All consecutive adults who were ventilated more than 48 h were included if VAP was clinically suspected. A dilution factor, k, was developed according to the formula: dilution factor k = concentration of urea in plasma/concentration of urea in lavage fluid recovered. Using this dilution factor, the quantitative bacterial counts were interpreted accordingly with a corrected positive threshold at 10(5) colony forming unit (CFU) mL(-1). MEASUREMENTS AND RESULTS: Eighty-nine BAL with at least one micro-organism > or = 10(4) CFU mL(-1) were identified (37%). In 176 BAL (73%), k ranged from 10 to 100. Median k was 24.4 (9.7-40.2) in VAP group and 24.6 (13.1-57.8) in patients without pneumonia (NS). Among the 25 BAL with micro-organism counts of 10(4) CFU mL(-1), 3 had a dilution factor lower than 10, resulting in corrected counts below the threshold of 10(5) CFU mL(-1). Two out of 15 patients with micro organism counts of 10(3) CFU mL(-1) had corrected micro-organism counts of 10(5) CFU mL(-1). Finally, only five BAL (2.1%) were misclassified when the dilution correction factor was applied. CONCLUSIONS: Using urea as dilution factor, we showed that BAL dilution variations did not alter the interpretation of BAL quantitative bacterial culture when administrating three aliquots of 50 mL of saline. PMID- 19189080 TI - Short-term evaluation of sedation with sevoflurane administered by the anesthetic conserving device in critically ill patients. AB - PURPOSE: Assessing feasibility and physiological effects of sedation with sevoflurane, administered with the anesthetic conserving device (AnaConDa), in comparison with propofol and remifentanil. METHODS: Seventeen patients undergoing mechanical ventilation underwent sedation with sevoflurane delivered with AnaConDa (phase SevAn), preceded and followed by sedation with propofol and remifentanil (phases ProRe(1), ProRe(2)), with the same sedation targets. RESULTS: With both strategies it was possible to achieve the sedation targets. Time required to sedate and awake patients was greater during SevAn than ProRe(1): respectively, 3.3 +/- 3.0 versus 8.9 +/- 6.1 and 7.47 +/- 5.05 versus 16.3 +/- 11.4 min. During SevAn the PaCO(2) and minute ventilation increased. Hemodynamics was stable between ProRe(1) and SevAn, except for an increase in heart rate in the SevAn phase. Environmental pollution from sevoflurane was within the safety limits. CONCLUSIONS: Sevoflurane can be effectively and safely used for short-term sedation of ICU patients with stable hemodynamic conditions. PMID- 19189081 TI - Relationship between gas exchange response to prone position and lung recruitability during acute respiratory failure. AB - PURPOSE: To clarify whether the gas exchange response to prone position is associated with lung recruitability in mechanically ventilated patients with acute respiratory failure. METHODS: In 32 patients, gas exchange response to prone position was investigated as a function of lung recruitability, measured by computed tomography in supine position. RESULTS: No relationship was found between increased oxygenation in prone position and lung recruitability. In contrast, the decrease of PaCO(2) was related with lung recruitability (R(2) 0.19; P = 0.01). Patients who decreased their PaCO(2) more than the median value (-0.9 mmHg) had a greater lung recruitability (19 +/- 16 vs. 8 +/- 6%; P = 0.02), higher baseline PaCO(2) (48 +/- 8 vs. 41 +/- 11 mmHg; P = 0.07), heavier lungs (1,968 +/- 829 vs. 1,521 +/- 342 g; P = 0.06) and more non-aerated tissue (1,009 +/- 704 vs. 536 +/- 188 g; P = 0.02) than those who did not. CONCLUSIONS: During prone position, changes in PaCO(2), but not in oxygenation, are associated with lung recruitability which, in turn, is associated with the severity of lung injury. PMID- 19189083 TI - Optimization of capillary electrophoresis conditions for a glucagon competitive immunoassay using response surface methodology. AB - The capillary electrophoresis (CE) conditions for a competitive immunoassay of glucagon were optimized for highest sensitivity of the immunoassay and resolution of the electrophoretic peaks using a Box-Behnken design. Injection time, voltage ramp time, and separation voltage were varied between three levels and two responses, bound-to-free (B/F) ratio of the immunoassay peaks and resolution between the peaks, were measured. Analysis of variance was applied to fit a predictive model, and a desirability function was used to simultaneously optimize both responses. A 10-s injection, 1.6-min ramp time, and a 22-kV separation voltage were the conditions found when high B/F was given more emphasis than high resolution. To test the model, calibration curves of a glucagon immunoassay were measured at the optimum and least optimum CE conditions. Optimal conditions increased the sensitivity of the immunoassay by 388% compared to the least optimum conditions while maintaining adequate resolution. PMID- 19189082 TI - Marble burying reflects a repetitive and perseverative behavior more than novelty induced anxiety. AB - RATIONALE: An increasing number of investigators utilize the marble-burying assay despite the paucity of information available regarding what underlies the behavior. OBJECTIVES: We tested the possibility that a genetic component underlies marble burying in mice and if there is a genetic correlation with other anxiety-like traits. Since findings reported in the literature indicate that marble-burying behavior reflects an anxiety-like response, we explored the assumption that the novel nature of a marble induces this anxiety. Finally, we investigated how the natural response of a mouse to dig relates to the marble burying phenomenon. METHODS: We examined ten different inbred mouse strains to determine if marble-burying behavior is genetically regulated and correlated with anxiety-like traits in two other assays. We employed multiple variants of the "traditional" marble-burying assay to address how issues such as the novelty of marbles and digging behavior contribute to marble burying. RESULTS: Marble burying behavior varied across strain and did not correlate with anxiety measures in other assays. Multiple tests conducted to reduce the novelty of marbles failed to alter burying behavior. Additionally, digging behavior correlated with marble burying, and the presence of marbles did not significantly impact the digging response. CONCLUSIONS: Our results indicate that mouse marble burying is genetically regulated, not correlated with other anxiety-like traits, not stimulated by novelty, and is a repetitive behavior that persists/perseveres with little change across multiple exposures. Marble burying is related to digging behavior and may in fact be more appropriately considered as an indicative measure of repetitive digging. PMID- 19189084 TI - Fast-scan cyclic voltammetry for the detection of tyramine and octopamine. AB - Tyramine and octopamine are biogenic amine neurotransmitters in invertebrates that have functions analogous to those of the adrenergic system in vertebrates. Trace amounts of these neurotransmitters have also been identified in mammals. The purpose of this study was to develop an electrochemical method using fast scan cyclic voltammetry at carbon-fiber microelectrodes to detect fast changes in tyramine and octopamine. Because tyramine is known to polymerize and passivate electrode surfaces, waveform parameters were optimized to prevent passivation. No fouling was observed for octopamine when the electrode was scanned from 0.1 to 1.3 V and back at 600 V/s, while a small decrease of less than 10% of the signal was seen for 15 repeated exposures to tyramine. The technique has limits of detection of 18 nM for tyramine and 30 nM for octopamine, much lower than expected levels in insects and lower than basal levels in some brain regions of mammals. Current was linear with concentration up to 5 microM. This voltammetry technique should be useful for measuring tyramine and octopamine changes in insects, such as the fruit fly, Drosophila melanogaster. PMID- 19189085 TI - Bio-inspired colorimetric detection of Hg2+ and Pb2+ heavy metal ions using Au nanoparticles. AB - Heavy metal ions are highly toxic species which can cause long-term damage to biological systems. These species are known to disrupt biological events at the cellular level, cause significant oxidative damage, and are carcinogens. The production of simple, in-field detection methods that are highly sensitive for these cations is highly desirable in response to global pollution. In that regard, bio-inspired colorimetric sensing systems have been developed to detect Hg(2+) and Pb(2+), and other cations, down to nmol L(-1) concentrations. The benefits of these systems, which are reviewed herein, include cost-effective production, facile usage, and a visual color change for the detection method. Such advantages are significant positive steps for heavy metal ion detection, especially in regions where sophisticated laboratory studies are prohibited. PMID- 19189086 TI - Cerebral cortical mechanisms of copying geometrical shapes: a multidimensional scaling analysis of fMRI patterns of activation. AB - We used multidimensional scaling (MDS) to characterize the integrative neural mechanisms during viewing and subsequently copying nine geometrical shapes. Human subjects initially looked at a central fixation point ("rest" period), then looked at a geometrical shape ("visual" period) which they copied without visual feedback ("copying" period). BOLD signal was recorded from voxels in 28 cortical areas (14 from each hemisphere) using a 4 Tesla magnet. For each voxel, signal ratios of "Visual versus Rest" (VR), and "Copy versus Visual" (CV) were calculated and used to construct two sets of Euclidean distance dissimilarity matrices for the nine shapes, with separate matrices defined for each region of interest (ROI) across subjects. The relations of perceptual and motor aspects of the shapes to MDS dimensions and specific ROIs were assessed using stepwise multiple regressions. The optimal individually scaled (INDSCAL) solutions were 2 dimensional. For the VR condition, MDS dimensions were significantly associated with the presence of crossing in a shape (Dimension 1), and with perimeter, height, cycles, peak segment speed, and horizontal symmetry (Dimension 2). ROIs most prominently associated with these dimensions essentially comprised the medial frontal lobe bilaterally, the inferior frontal gyrus bilaterally, and the left intraparietal sulcus (Dimension 1), and visual areas, including the calcarine sulcus and cuneus bilaterally (Dimension 2). These results document the expected involvement of visual areas and support the hypothesis advanced on the basis of previous findings (Lewis et al. 2003a) that a motor rehearsal of the upcoming shape copying is occurring during this visual presentation period. For the CV condition, practically one motor feature (number of segments drawn) dominated both dimensions, with a secondary engagement of horizontal symmetry in Dimension 1. The right postcentral gyrus, right intraparietal sulcus, right superior parietal lobule and right inferior parietal lobule contributed mostly to Dimension 1; the superior frontal gyrus bilaterally, right middle frontal gyrus, left postcentral gyrus, left inferior parietal lobule contributed mostly to Dimension 2; and the left superior parietal lobule and left intraparietal sulcus contributed to both dimensions approximately equally. CV BOLD activation of ROIs contributing to Dimension 1 (or to both dimensions) was significantly associated with the number of shape segments drawn. Since the direction of movement differs in successively drawn shape segments, the number of segments (minus one) equals the number of changes in the direction of movement. We conclude that this fundamental spatial motor aspect of drawing geometrical shapes is the critical variable, independent of the particular shape drawn, that dominates cortical activation during copying. PMID- 19189087 TI - Withdrawal reflex responses evoked by repetitive painful stimulation delivered on the sole of the foot during late stance: site, phase, and frequency modulation. AB - The modulation of the lower limb nociceptive withdrawal reflex elicited during late stance by a stimulus train with frequencies of 15 and 30 Hz delivered to the mid-forefoot, arch of the foot, and heel was investigated. Stimulation was delivered at four moments of the gait cycle between heel-off and toe-off. Stimulation at 15 Hz induced larger kinematic responses at the knee and hip. Reduced plantarflexion and increased dorsiflexion, compared to control steps, were evoked at the ankle; these kinematic responses were site dependent with minimum responses evoked by stimulation at the heel. The dorsiflexion response was largest when stimulating at toe-off and was larger for stimulation at 15 Hz than at 30 Hz. The muscle reflex responses were site modulated in tibialis anterior with largest responses evoked by stimulation at the arch of the foot and mid-forefoot, and phase and frequency modulated in soleus. This study presents a detailed assessment of the lower limb nociceptive reflex modulation and provides results, which might have application in the rehabilitation of the hemiparetic gait. PMID- 19189088 TI - The bioavailability of bromazepam, omeprazole and paracetamol given by nasogastric feeding tube. AB - AIMS: To characterize and compare the pharmacokinetic profiles of bromazepam, omeprazole and paracetamol when administered by the oral and nasogastric routes to the same healthy cohort of volunteers. METHODS: In a prospective, monocentric, randomized crossover study, eight healthy volunteers received the three drugs by the oral (OR) and nasogastric routes (NT). Sequential plasma samples were analyzed by high-performance liquid chromatography-UV, pharmacokinetic parameters (Cmax, AUC(0-infinity), t(1/2), k(e), tmax) were compared statistically, and Cmax, AUC(0-infinity) and t(max) were analyzed for bioequivalence. RESULTS: A statistically significant difference was seen in the AUC(0-infinity) of bromazepam, with nasogastric administration decreasing availability by about 25%: AUC(OR) = 2501 ng mL(-1) h; AUC(NT) = 1855 ng mL(-1) h (p < 0.05); ratio (geometric mean) = 0.74 [90% confidence interval (CI) 0.64-0.87]. However, this does not appear to be clinically relevant given the usual dosage range and the drug's half-life (approx. 30 h). A large interindividual variability in omeprazole parameters prevented any statistical conclusion from being drawn in terms of both modes of administration despite their similar average profile: AUC(OR) = 579 ng mL(-1) h; AUC(NT) = 587 ng mL(-1) h (p > 0.05); ratio (geometric mean) = 1.01 (90% CI 0.64-1.61). An extended study with a larger number of subjects may possibly provide clearer answers. The narrow 90% confidence limits of paracetamol indicate bioequivalence: AUC(OR) = 37 microg mL(-1) h; AUC(NT) = 41 microg mL(-1) h(p > 0.05); ratio (geometric mean) = 1.12 (90% CI 0.98-1.28). CONCLUSION: The results of this study show that the nasogastric route of administration does not appear to cause marked, clinically unsuitable alterations in the bioavailability of the tested drugs. PMID- 19189089 TI - A study of the structure of the crystalline bacterial biofilms that can encrust and block silver Foley catheters. AB - The aim of this study was to examine the structure of the crystalline bacterial biofilms that encrust and block silver/hydrogel-coated latex catheters. Scanning electron microscopy was used to examine the crystalline deposits that were found encrusting catheters obtained from six patients undergoing long-term catheterization in a community setting. Large populations of bacilli and cocci were seen on all catheters developing on a basal foundation layer of crystalline material. These observations show that in patients prone to catheter encrustation, crystalline material formed in the urine can cover the surfaces of silver catheters. Extensive bacterial biofilms then develop on the crystals, shielded from the underlying silver. It is suggested that if antimicrobials are to be incorporated into catheters to prevent encrustation, they must diffuse out from the catheter surface and reduce the viable cell populations of the urease producing bacteria that elevate the urinary pH and trigger crystal formation. PMID- 19189090 TI - Ovarian apoptosis after shock wave lithotripsy for distal ureteral stones. AB - The objective of this study is to identify any apoptotic effect of shock wave lithotripsy (SWL) for distal ureteral stones on ovarian tissue. Twenty-one female New Zealand White rabbits were divided into three groups of seven rabbits each: I (control), and II, III (treated and killed 14 and 28 days after SWL, respectively). The left distal ureteral segment of the anesthetized (ketamine HCl, 20 mg/kg) animals in groups II and III was exposed to 1,500 shock waves at 17 kV. Localization of the distal ureteral segments was achieved following contrast medium (Iohexol 300 mg of I/ml) injection. The animals were killed on day 14 or 28 after SWL, and the ovaries were removed. The follicle number with apoptotic changes in ovarian tissue was compared with control group. Apoptotic changes were determined by terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end-labeling (TUNEL) method. No increased apoptosis was detected in all groups. The mean number of TUNEL-positive follicle in groups I, II and III was 9.3 +/- 2.9, 8.1 +/- 2.6 and 8.7 +/- 2.9, respectively. There were no statistically significant differences among all groups regarding the number of TUNEL-positive follicle (P = 0.647). Also, no histomorphological change other than apoptosis was detected in the study groups. In conclusion, SWL treatment for distal ureteral stones does not induce apoptotic changes on ovarian tissue. PMID- 19189091 TI - Building a culture of research among clinical pediatric radiologists: a multifaceted, programmatic approach. PMID- 19189092 TI - Evaluation of the anomalous retroesophageal left brachiocephalic vein in Chinese children using multidetector CT. AB - BACKGROUND: Anomalous retroesophageal left brachiocephalic vein is a very rare anomaly of the brachiocephalic vein. At the time of writing only one case had been reported worldwide. OBJECTIVE: The purpose of this study was to report four cases of anomalous retroesophageal left brachiocephalic vein and to evaluate the utility of multidetector computed tomography (MDCT) for investigation of the anomaly. MATERIALS AND METHODS: From June 2005 to May 2008, we examined 2,146 children with congenital heart disease and 4,082 children without congenital heart disease using MDCT. The patients ranged in age from 1 day to 12 years. Maximum intensity projection reconstruction was performed to show the brachiocephalic vein. RESULTS: A retroesophageal left brachiocephalic vein was found in 4 of the 2,146 patients (0.19%) with congenital heart disease and in 0 of the 4,082 patients without congenital heart disease. In all four patients, the condition was confirmed during surgery for the congenital heart disease. CONCLUSION: MDCT is a good modality for evaluating the anomalous retroesophageal left brachiocephalic vein. An anomalous retroesophageal left brachiocephalic vein was more common in patients with congenital heart disease. PMID- 19189093 TI - Perflubron residua: 12 years following therapy. AB - Partial liquid ventilation therapy utilizing perfluorocarbons has been used for patients with severe respiratory distress. Perfluorocarbons such as perflubron have been demonstrated to clear from the lungs shortly after therapy. We present a 13-year-old boy with residual perflubron in his lungs, 12 years following therapy with perflubron. The imaging features are important to recognize as these patients return for additional care and imaging. PMID- 19189094 TI - CT findings in children with Meckel diverticulum. AB - BACKGROUND: Approximately 0.04% of the general population will present with a complication related to Meckel diverticulum. The classic teaching is that symptomatic children with Meckel diverticulum present with painless rectal bleeding and are evaluated with a radionuclide scan. Our subjective experience is that we see children with Meckel diverticulum who present with abdominal pain and are evaluated by CT. OBJECTIVE: We reviewed the findings on CT in children with pathologically proven Meckel diverticulum to identify characteristic patterns of presentation. MATERIALS AND METHODS: Databases were searched (2004-2008) for all children who had a pathologic diagnosis of Meckel diverticulum and a CT scan performed prior to surgery. Demographics, pathology, and CT features were reviewed. CT features reviewed included: soft-tissue stranding, abnormal calcifications, bowel obstruction, free air, free peritoneal fluid, cystic mass, intussusception, obvious lead point, location, and whether a normal appendix was identified. The frequency of Meckel diverticulum encountered on CT scans was compared to that found during the same period of time on technetium pertechnetate studies. RESULTS: The review identified 16 subjects (mean age 9.5 years, M:F 9:7). CT findings included: soft-tissue stranding in nine (56%), small-bowel obstruction (SBO) in nine (56%), intussusception in three (19%), free fluid in ten (63%), cystic mass in four (25%), calcification in none (0%), free air in one (6%), and no abnormalities in two (13%). A normal appendix was identified in only five children (31%). There were three basic patterns of presentation of abnormalities: SBO only in five, intussusception with SBO in three, or cystic mass with inflammatory stranding in four (one with SBO). Also, 2.3 times more Meckel diverticulum was encountered on CT than on technetium pertechnetate studies. CONCLUSION: Meckel diverticulum is currently more commonly encountered in children on CT performed for abdominal pain than on technetium pertechnetate studies. There are three categories of appearance on CT: SBO only, intussusception, or a cystic inflammatory mass. PMID- 19189095 TI - Proximal humeral epiphysiolysis in a newborn. PMID- 19189096 TI - Scrotal emergencies. AB - This review describes the clinical, grey-scale and colour Doppler US findings of different conditions that may be the cause of an acute scrotum. The US appearances of testicular torsion, torsion of the appendix testis, epididymoorchitis, incarcerated hernia and idiopathic scrotal oedema are described. Some of the major studies regarding the sensitivity and specificity of US in the diagnosis of these diseases are reviewed. Grey-scale and colour Doppler US demonstrate high accuracy in the aetiological diagnosis of an acute scrotum and can differentiate between diseases that require immediate surgical intervention and those that can be treated conservatively. PMID- 19189097 TI - Imaging of acute appendicitis in children: EU versus U.S. ... or US versus CT? A European perspective. AB - There is substantial evidence that imaging may reduce the negative appendectomy rate, also in children. However, controversy exists about the preferred method: US or CT, and the choice appears to be determined by the side of the Atlantic Ocean. This review brings forth several arguments in favour of US. PMID- 19189098 TI - CT findings in unilateral hepatopulmonary syndrome after the Fontan operation. AB - BACKGROUND: Patients with complex congenital heart defects palliated by connecting the systemic veins directly to the pulmonary circulation are known to develop hepatopulmonary syndrome (HPS). Although rare, HPS can develop following the Fontan operation. OBJECTIVE: To present and analyse the CT findings of HPS in patients with a Fontan circulation. MATERIALS AND METHODS: From May to December 2005, six patients with HPS following the completion of a Fontan circulation were evaluated. CT findings were reviewed and were compared with angiographic findings. RESULTS: All six patients showed unilateral involvement. All patients except one had inferior vena cava (IVC) interruption with azygos continuation. CT scans showed abnormal vascular dilatation in one lung, and properly demonstrated the anatomy causing the hepatic venous blood to flow preferentially into one lung. These CT findings correlated well with the angiography findings. CONCLUSION: HPS that develops after the Fontan procedure is typically unilateral and is often associated with IVC interruption and azygos or hemiazygos continuation. CT demonstrates dilatation of pulmonary vessels in the affected lung and may be able to demonstrate the underlying anatomical cause for the predilection of hepatic venous flow to the contralateral lung. PMID- 19189100 TI - 17th international symposium on analytical ultracentrifugation and hydrodynamics. PMID- 19189099 TI - Neonatal colour Doppler ultrasound study: normal values of abdominal blood flow velocities in the neonate during the first month of life. AB - BACKGROUND: Doppler US to measure abdominal blood flow velocities (ABFV) is increasingly used to investigate intestinal haemodynamics in several clinical conditions in neonates. Studies that provide reference values of ABFV during the entire neonatal period are currently lacking. OBJECTIVE: To make available normal reference values of ABFV and Doppler indices in the coeliac trunk and superior mesenteric artery during the first month of life in term and healthy preterm infants. MATERIALS AND METHODS: ABFV were obtained with colour Doppler US in 69 neonates (12 term, 57 preterm) divided into four gestational age groups (25-28 weeks, 29-32 weeks, 33-36 weeks, and 37-41 weeks). RESULTS: ABFV increased with increasing gestational and postnatal age. We also provide normal reference values of ABFV and Doppler indices to compare with measurements of abdominal blood flow changes during the neonatal period for diagnostic, therapeutic and prognostic purposes. CONCLUSION: These longitudinal reference values provide a useful tool for assessing possible alteration in ABFV secondary to neonatal pathologies. PMID- 19189101 TI - Insights into protein-polysorbate interactions analysed by means of isothermal titration and differential scanning calorimetry. AB - Therapeutic proteins formulated as liquid solutions at high protein concentration are very sensitive to chemical and physical degradation. Especially avoiding the formation of protein aggregates is very crucial for product quality. In order to stabilize the colloidal properties of protein therapeutics various excipient are used. Especially the detergents polysorbate 20 and 80 are common. However, the mechanism upon which the detergents protect the protein from aggregation is not really known. The present study investigates the interaction of polysorbate 20 and 80 with different proteins: lysozyme, bovine serum albumin (BSA) and an immunoglobulin. The interaction and binding of the detergents to the proteins is investigated by isothermal titration calorimetry (ITC). From ITC the thermodynamic parameters (DeltaH: change in enthalpy, DeltaS: entropy and DeltaG: free energy) upon binding are derived as well as the binding constant K (a). The thermal stability of the proteins in the presence of the detergent is assessed by differential scanning calorimetry (DSC). The results show that both detergents bind to BSA with K (a) between 8 and 12 x 10(3) M(-1) with DeltaH -50 to -60 kJ/mol (25 degrees C). One to two detergent molecules bind to BSA. The presence of both detergents induces a weak stabilisation of the thermal denaturation properties of BSA. However, the interaction of polysorbate 20 and 80 with lysozyme and the immunoglobulin is quite negligible. The presence of the detergents up to a concentration of 2 mM has no impact on the heat capacity curve neither a destabilisation nor a stabilisation of the native conformation is observed. PMID- 19189102 TI - Dense chromatin plates in metaphase chromosomes. AB - In a previous work we observed multilayered plate-like structures surrounding partially denatured HeLa chromosomes at metaphase ionic conditions. This unexpected finding has led us to carry out an extensive investigation of these structures. Our results show that plates can also be found in metaphase chromosomes from chicken lymphocytes. We have used atomic force microscopy (AFM) to image and investigate the mechanical properties of plates in aqueous solution. Plates are thin (approximately 6.5 nm each layer) but compact and resistant to penetration by the AFM tip: their Young's modulus is approximately 0.2 GPa and the stress required for surface penetration is approximately 0.03 GPa in the presence of Mg(2+) (5-20 mM). Low-ionic strength conditions produce emanation of chromatin fibers from the edges of uncrosslinked plates. These observations and AFM results obtained applying high forces indicate that the chromatin filament is tightly tethered inside the plates. Images of metal-shadowed plates and cryo electron microscopy images of frozen-hydrated plates suggest that nucleosomes are tilted with respect to the plate surface to allow an interdigitation between the successive layers and a thickness reduction compatible with the observed plate height. The similarities between denatured plates from chicken chromosomes and aggregates of purified chromatin from chicken erythrocytes suggest that chromatin has intrinsic structural properties leading to plate formation. Scanning electron micrographs and images obtained with the 200-kV transmission microscope show that plates are the dominant component of compact chromatids. We propose that metaphase chromosomes are formed by many stacked plates perpendicular to the chromatid axis. PMID- 19189103 TI - TCR transgenes and transgene cassettes for TCR gene therapy: status in 2008. AB - The genetic introduction of T cell receptor genes into T cells has been developed over the past decade as a strategy to induce defined antigen-specific T cell immunity. With the potential value of TCR gene therapy well-established in murine models and the feasibility of infusion of TCR-modified autologous T cells shown in a first phase I trial, the next key step will be to transform TCR gene transfer from an experimental technique into a robust clinical strategy. In this review, we discuss the different properties of the TCR transgene and transgene cassette that can strongly affect both the efficacy and the safety of TCR gene transfer. PMID- 19189105 TI - The addition of radiotherapy to chemotherapy does not improve outcome of early stage Hodgkin's lymphoma patients: a retrospective long-term follow-up analysis of a regional Italian experience. AB - We retrospectively reviewed 139 stage I-II HL patients who were diagnosed and followed up in an Italian northern region (Liguria) from 1995 to 2007, and who received either chemotherapy (CT) alone (mainly doxorubicin, bleomycin, vinblastine, and dacarbazine; ABVD) or a combined modality treatment (chemotherapy + radiotherapy, CT + RT). The two therapeutic groups were comparable for clinical and histologic features. Complete remission rate after CT + RT was higher than what was achieved with CT alone (96% vs. 84%, respectively, p = 0.03). Relapse rate (12%) was the same in both groups and disease-free survival curves were comparable (82% and 83%, p = 0.47). The overall survival of the two therapeutic groups is comparable. No second tumors have been reported among patients receiving chemotherapy alone, whereas a second neoplasia has been diagnosed in four patients (in two cases possibly radiotherapy related) in the CT + RT group (5%, p = 0.09) In conclusion, our retrospective study shows that CT + limited RT is an effective and well-tolerated option for early stage Hodgkin's lymphoma, even if the use of RT is associated with a certain risk of developing a second tumor. However, four to six courses of ABVD can lead to similar, optimal, long-term disease control without exposing patients to the risk of a second neoplasia. PMID- 19189104 TI - Stability of medial opening wedge high tibial osteotomy: a failure analysis. AB - Medial opening wedge high tibial osteotmy (HTO) is often used to treat varus gonarthrosis in young, active, highly demanding patients, although it has many pitfalls, which were evaluated in a consecutive cohort of patients. A retrospective analysis of a consecutive series of 45 patients with 49 medial opening HTO for varus gonarthrosis using a spacer plate (Puddu I, Arthrex, USA) were included. A Chi square test was used to study the effect between the wedge size and complications. Complications occurred in 22 knees (45%). There was no significant difference between groups for individual complications; however, when combined, there were significantly more complications in the >10 mm wedge group (Chi square p = 0.05). The overall complication rate in this series was 45%. The majority were related to intrinsic instability at the osteotomy site (24%) and surgical technique (20%). The evaluated spacer provided inadequate stability. PMID- 19189106 TI - Oxaliplatin, irinotecan, and fluorouracil/folinic acid in advanced gastric cancer: a multicenter phase II trial of the Southern Italy Cooperative Oncology Group. AB - PURPOSE: This phase II trial assessed the tolerability and efficacy of a triplet of oxaliplatin, irinotecan, and fluorouracil/folinic acid in advanced gastric cancer. METHODS: Patients with unresectable or metastatic gastric cancer, unexposed to palliative chemotherapy, received oxaliplatin 85 mg/m(2) iv and irinotecan 150 mg/m(2) iv on day 1, 6S-folinic acid 250 mg/m(2) iv and fluorouracil 750 mg/m(2) iv on day 2, every 2 weeks. Response rate (RR) was assessed after a minimum of four cycles, and treatment continued up to 12 cycles. RESULTS: Sixty-three patients were treated, with a median of eight (range 1-12) cycles/patient. Two complete and 19 partial responses were registered (RR 33% [95% CI, 22-46%]). Median progression-free survival was 7.5 (95% CI, 5.6-9.4) months, and median overall survival was 12.1 (95% CI, 10.8-13.4) months. Most common grade > or =3 toxicities were neutropenia (59%), febrile neutropenia (7%), vomiting (20%), and diarrhoea (10%). All-grade neurotoxicity affected 33% of patients. CONCLUSIONS: Oxaliplatin, irinotecan, and fluorouracil/folinic acid administered every 2 weeks are safe and active in advanced gastric cancer. PMID- 19189107 TI - Modeling the mechanism of postantibiotic effect and determining implications for dosing regimens. AB - A stochastic model is proposed to explain one possible underlying mechanism of the postantibiotic effect (PAE). This phenomenon, of continued inhibition of bacterial growth after removal of the antibiotic drug, is of high relevance in the context of optimizing dosing regimens. One clinical implication of long PAE lies in the possibility of increasing intervals between drug administrations. The model describes the dynamics of synthesis, saturation and removal of penicillin binding proteins (PBPs). High fractions of saturated PBPs are in the model associated with a lower growth capacity of bacteria. An analytical solution for the bivariate probability of saturated and unsaturated PBPs is used as a basis to explore optimal antibiotic dosing regimens. Our finding that longer PAEs do not necessarily promote for increased intervals between doses, might help for our understanding of data provided from earlier PAE studies and for the determination of the clinical relevance of PAE in future studies. PMID- 19189108 TI - Diffusion tensor tractography of the somatosensory system in the human brainstem: initial findings using high isotropic spatial resolution at 3.0 T. AB - Lack of adequate sensitivity and spatial resolution in previous noninvasive imaging studies has impeded the depiction of different somatosensory pathways (the medial lemniscus and spinal lemniscus). We investigated whether incorporation of diffusion tensor imaging (DTI) at high isotropic spatial resolution and DTI-based 3D fiber-tractography information can facilitate the study of anatomical parcellation of the somatosensory system in the healthy adult human brainstem. Five healthy men (age range 24-37 years) were studied, and written informed consent was obtained from all subjects. Three-Tesla MRI diffusion tensor tractography (DTT) using fiber assignment by the continuous tracking (FACT) approach at high spatial resolution was used to reconstruct three white matter tracts, the medial lemniscus (ML), spinal lemniscus (SL), and central tegmental tract (CTT), to delineate and quantify the sensory pathways within the brainstem. We demonstrate that these three pathways are distinguishable from each other. The tractographic patterns of the three pathways on all subjects were similar and consistent with atlases of anatomy. We also quantified the diffusion tensor metrics (fractional anisotropy and mean diffusivity) of the two somatosensory pathways, the SL and ML. The fractional anisotropy of the ML was significantly higher than that of the SL (p = 0.005) The average diffusivity was significantly smaller for the ML than for the SL (p = 0.003). PMID- 19189109 TI - Magnetic resonance imaging for evaluation of disease activity in Crohn's disease: a systematic review. AB - To systematically review the evidence on the accuracy of MRI for grading disease activity in Crohn's disease (CD). The MEDLINE, EMBASE, CINAHL and Cochrane databases were searched for studies on the accuracy of MRI in grading CD compared to a predefined reference standard. Two independent observers scored all relevant data. Three disease stages were defined: remission, mild and frank disease. The accuracy rates of MRI per disease stage were calculated by means of a random effects model. Seven studies were included from a search resulting in 253 articles. In total 140 patients (16 patients in remission, 29 with mild disease and 95 with frank disease) were used for data analysis. MRI correctly graded 91% (95% CI: 84-96%) of patients with frank disease, 62% (95% CI: 44-79) of patients with mild disease and 62% (95% CI: 38-84) of patients in remission. MRI more often overstaged than understaged disease activity; MRI overstaged disease activity in 38% of patients in remission, mostly as mild disease. Overstaging of mild disease was observed in 21%, understaging in 17%. MRI correctly grades disease activity in a large proportion of patients with frank disease. For patients in remission or with mild disease, MRI correctly stages disease activity in many patients (62%). PMID- 19189110 TI - Long-term functional results following resection of neonatal sacrococcygeal teratoma. AB - PURPOSE: Sacrococcygeal teratoma (SCT) is the most common congenital neoplasm in neonates. We wished to assess the long-term functional outcome of children undergoing SCT resection. METHODS: Records of neonates diagnosed with SCT from two surgeons' practices, and operated on between 1970 and 2006, were retrospectively reviewed. Patients/parents who consented to participate in the study received a questionnaire, focusing on fecal and urinary continence, constipation and lower extremity weakness. RESULTS: Forty-six patients were identified. Four had died (3 from malignant tumors and 1 motor vehicle accident at 18 years). Of the 42 remaining cases, 39 were benign and 3 were malignant; 2 of the former developed malignant recurrences. Twenty-seven agreed to participate and 14 (52%) completed the questionnaire. Median age of respondents was 16.7 years (3-29), and none of the respondents had a recurrent tumor. Thirteen of the 14 respondents experienced no problem with urinary or fecal incontinence, or lower extremity weakness. The remaining patient had all three problems, but his SCT had involved the spinal cord. Of the ten patients who commented on constipation, one had significant constipation, five occasional constipation, and four no constipation. CONCLUSIONS: Functional results after resection of neonatal SCT are excellent, with only a small number of patients reporting problems with fecal or urinary continence, or lower extremity weakness. Constipation is relatively common. This information is important for counseling families with fetal or neonatal SCT. PMID- 19189111 TI - The value of sacral nerve stimulation in the treatment of faecal incontinence after pelvic radiotherapy. PMID- 19189112 TI - The stability of impacted morsellized bone grafts in a metal cage under dynamic loaded conditions: an in vitro reconstruction of a segmental diaphyseal bone defect. AB - INTRODUCTION: Reconstructions of segmental diaphyseal bone defects with massive allografts are related to complications like nonunion and fractures. A reconstruction of these defects with a cage filled with an impacted morsellized bone graft could be an alternative. The bone graft in these cages should ideally be loaded to prevent resorption. Loading of morsellized bone grafts however can cause instability. The goal of this study was to assess the stability of an impacted morsellized bone graft in a cage under dynamic loaded conditions in an in vitro reconstruction of a segmental diaphyseal bone defect. The second goal was to assess the influence of cage type, washing of the graft and bone-cage fit. MATERIALS AND METHODS: Two different cage types were filled with impacted morsellized bone graft. The grafts were used washed and unwashed and in variable bone-cage fit conditions. We recorded the bone graft deformation in the cage under dynamic loaded conditions. RESULTS: Stability appeared to be not very sensitive to the cage type and whether the bone chips were washed or not. However, quality of the fit of the cage with the bone segment was an important parameter and should be optimized during surgery. CONCLUSIONS: Morsellized impacted bone graft in a cage is stable in dynamic loaded conditions in an in vitro reconstruction of a segmental diaphyseal bone defect. We believe that this method of reconstruction is a promising alternative for the reconstruction of large diaphyseal bone defects and should be tested relative to its biological merits in animal experiments. PMID- 19189113 TI - Bacteriology of postradiotherapy chronic rhinosinusitis in nasopharyngeal carcinoma patients and chronic rhinosinusitis. AB - The purpose of this study was to investigate and compare the bacteriology of postradiotherapy chronic rhinosinusitis (postRT-CRS) and chronic rhinosinusitis (CRS) by evaluating the aspiration materials of the maxillary sinus of patients with postRT-CRS and patients with CRS. We collected the secretions of the maxillary sinus from 30 nasopharyngeal carcinoma patients with postRT-CRS and 30 patients with CRS for aerobe/facultative anaerobe bacteria culture. The most common isolates in the postRT-CRS group were Streptococcus viridans, Staphylococcus aureus and Haemophilus influenzae, while those in the CRS group were Haemophilus influenzae, Pseudomonas aeruginosa and Staphylococcus aureus. Isolated gram-positive coccus rate in postRT-CRS patients was significantly higher than in CRS patients (62.50% compared with 30.00%, respectively; P < 0.05), and isolated gram-negative bacilli rate in postRT-CRS patients was significantly lower than in CRS patients (31.25% compared with 70.00%, respectively; P < 0.05). However, the incidence of positive cultures was not significantly different between the postRT-CRS group and the CRS group (P > 0.05). This study found that there were some differences in bacteriology between postRT-CRS and CRS. Gram-positive coccus was the predominant aerobic/facultative anaerobe pathogenic bacterium in patients with postRT-CRS, and gram-negative bacilli was predominant in CRS patients. PMID- 19189114 TI - Radiofrequency thermal ablation for patients with nasal symptoms: a systematic review of effectiveness and complications. AB - Radiofrequency ablation (RFA) is a relatively new method for the reduction of submucosal tissue. The method has gained increasing popularity in the treatment of snoring, tonsillar hypertrophy, tongue base hypertrophy, and nasal obstruction secondary to non-allergic or allergic rhinitis. We present a systematic literature review of current knowledge on the effectiveness, and complications of nasal radiofrequency thermal ablation (RFA) in the treatment of nasal obstruction. We performed a computerized literature search using several databases to select articles dealing with RFA treatment in the field of otorhinolaryngology. Selected articles were independently appraised by at least two of the authors. Thirty-five articles met the inclusion criteria. Twenty-six articles were reports on uncontrolled patient series. Of the nine included randomized controlled trials only one reported a double-blind comparison. Most of the studies reported an improvement in subjective symptoms after treatment and the number of serious side effects was small. However, the only placebo controlled trial did not show effectiveness. Nasal-RFA appears to be a safe operative procedure and may reduce inferior turbinate submucosal tissue volume in patients having chronic nasal obstruction and who fail to respond to medical treatment. Based on current knowledge, RFA alters the nasal mucosa only slightly and causes only minor discomfort and risk of side effects for the patient. However, most of the published studies on nasal-RFA are observational and have a relatively short follow-up. Consequently, there is an urgent need for well planned, double-blind, placebo-controlled randomized trials on nasal-RFA treatment. PMID- 19189117 TI - Kestenbaum procedure with posterior fixation suture for anomalous head posture in infantile nystagmus. AB - BACKGROUND: The purpose of this study was to report the effect of combining the Kestenbaum procedure with posterior fixation suture for infantile horizontal nystagmus with anomalous head posture (AHP) in children. METHODS: Nine consecutive patients who underwent combined Kestenbaum procedure plus posterior fixation suture to the recessed muscles at the same time were retrospectively studied. All patients were orthotropic before surgery and were followed for at least 6 months. Pre- and postoperative AHP and binocular corrected visual acuity (BCVA), and ocular alignment were assessed. RESULTS: Mean age at surgery was 4.8 +/- 1.5 years. The average follow-up was 29.7 months. The average head turn preoperatively was 27.4 degrees and postoperatively 7.2 degrees . The average net change in AHP was 24.8 degrees (P = 0.008). Seven of 9 patients (78%) achieved a residual head turn of 10 degrees or less. The average Log Mar BCVA was 0.33 preoperatively and 0.31 postoperatively (P = 0.68). Only 1 patient needed additional surgery for residual horizontal AHP. No patient developed strabismus. CONCLUSION: Combined Kestenbaum procedure with posterior fixation suture was an effective and stable procedure in reducing AHP of the range of 20 degrees to 35 degrees in children with infantile nystagmus. PMID- 19189116 TI - Triamcinolone acetonide prevents oxidative stress-induced tight junction disruption of retinal pigment epithelial cells. AB - PURPOSE: Oxidative stress is known to disrupt the integrity of retinal pigment epithelium (RPE) tight junctions. The goal of this study is to evaluate the effect of triamcinolone acetonide (TA) on the junctional integrity of RPE under oxidative stress and to identify the underlying mechanisms. METHODS: Second passage porcine RPE cells were cultured on 6-well membrane inserts until 4 weeks after reaching confluence. Cells were incubated with TA (10(-5) M) for 30 min. FITC-containing medium was added to the upper chamber (cell's apical side). The cells were then challenged with 1 mM Hydrogen Peroxide (H(2)O(2)). After 5 h, the fluorescence intensity of the medium from lower chamber (cell's basolateral side) was measured using a fluorescence spectrofluorophotometer. This transepithelial flux of FITC-dextran was measured until the 21st day. The immunolocalization of occludin and F-actin was examined with fluorescence microscope. Reduced glutathione (GSH)/oxidized glutathione (GSSG) ratio was determined by a colorimetric assay kit. RESULTS: Non-lethal oxidative stress by H(2)O(2) increased transepithelial flux of FITC-dextran significantly. TA inhibited this increase and preserved the lower flux through the whole experimental period. This permeability change by H(2)O(2) was reversible and recovered to the normal level within 3 weeks. In immunohistological study, H(2)O(2) reduced linear occludin staining at the cell border and increased actin stress fibers. TA prevented H(2)O(2)-induced disruption of junctional assembly of occludin and F-actin. Glutathione assay demonstrated that intracellular GSH/GSSG ratio decreased significantly with H(2)O(2), while TA preserved this ratio by up-regulating GSH synthesis. CONCLUSIONS: TA has a protective effect against oxidative stress induced disruption of RPE tight junction by preserving cellular redox state. PMID- 19189120 TI - Neural coding and contextual influences in the whisker system. AB - A fundamental problem in neuroscience, to which Prof. Segundo has made seminal contributions, is to understand how action potentials represent events in the external world. The aim of this paper is to review the issue of neural coding in the context of the rodent whiskers, an increasingly popular model system. Key issues we consider are: the role of spike timing; mechanisms of spike timing; decoding and context-dependence. Significant insight has come from the development of rigorous, information theoretic frameworks for tackling these questions, in conjunction with suitably designed experiments. We review both the theory and experimental studies. In contrast to the classical view that neurons are noisy and unreliable, it is becoming clear that many neurons in the subcortical whisker pathway are remarkably reliable and, by virtue of spike timing with millisecond-precision, have high bandwidth for conveying sensory information. In this way, even small (approximately 200 neuron) subcortical modules are able to support the sensory processing underlying sophisticated whisker-dependent behaviours. Future work on neural coding in cortex will need to consider new findings that responses are highly dependent on context, including behavioural and internal states. PMID- 19189119 TI - The role of the menstrual cycle phase in pain perception before and after an isometric fatiguing contraction. AB - The purpose of this study was to compare exercise-induced analgesia in young women after a fatiguing isometric contraction during different phases of the menstrual cycle. Twenty female subjects performed a submaximal (25% maximal voluntary contraction) isometric contraction until task failure during both the mid-follicular and mid-luteal phases of their menstrual cycle. Pain perception (i.e., pain threshold and pain ratings) was measured before and after the isometric fatiguing contraction. Other measures included mean arterial pressure, heart rate, and anxiety levels. Time to task failure of the fatiguing contraction was similar for the two phases of the menstrual cycle. Following the performance of the isometric contraction: (1) pain thresholds increased and pain ratings decreased; (2) anxiety levels increased; and (3) mean arterial pressure and heart rate increased. These changes were not dependent on the phase of the menstrual cycle. Thus, the menstrual cycle phase does not influence the magnitude of exercise-induced analgesia. PMID- 19189118 TI - Comparison of two doses of primary intravitreal bevacizumab (Avastin) for diffuse diabetic macular edema: results from the Pan-American Collaborative Retina Study Group (PACORES) at 12-month follow-up. AB - BACKGROUND: To report the 12-month anatomic and ETDRS best-corrected visual acuity (BCVA) response after primary intravitreal bevacizumab (Avastin) (1.25 mg or 2.5 mg) in patients with diffuse diabetic macular edema (DDME). In addition, a comparison of the two different doses of intravitreal bevacizumab (IVB) utilized was made. METHODS: We reviewed the clinical records of 82 consecutive patients (101 eyes) with DDME in this interventional retrospective multicenter study. All patients with a minimum follow-up of 12 months (mean 57.6 +/- 8.4 weeks) were included in this analysis. Patients underwent ETDRS best-corrected visual acuity (BCVA) testing, ophthalmoscopic examination, optical coherence tomography (OCT), and fluorescein angiography (FA) at baseline and follow-up visits. RESULTS: The mean age of our patients was 59.7 +/- 9.3 years. The mean number of IVB injections per eye was three (range: one to six injections) at a mean interval of 14.1 +/- 10.5 weeks. In the 1.25 mg group at 1 month BCVA improved from 20/190, logMAR = 0.97 to 20/85, logMAR 0.62, a difference that was statistically significant (p = 0.0001). This improvement was maintained throughout the 3-, 6-, and 12-month follow-up. The mean final BCVA at 12 months was 20/76, logMAR = 0.58 (p < 0.001), a statistically significant difference from baseline BCVA. Similar BCVA changes were observed in the 2.5 mg group. In the 1.25 mg group, the mean central macular thickness (CMT) decreased from 419.1 +/- 201.1 microm at baseline to 295.11 +/- 91.5 microm at 1 month, 302.1 +/- 124.2 microm at 3 months, 313.4.1 +/- 96.3 microm at 6 months, and 268.2 +/- 95.5 microm at 12 months (p < 0.0001). Similar CMT changes were observed in the 2.5 mg group. Adverse events included transient high blood pressure in one patient (1.2%), transient increased intraocular pressure in one eye (1%), and tractional retinal detachment in one eye (1%). CONCLUSIONS: Primary IVB at doses of 1.25 to 2.5 mg seem to provide stability or improvement in BCVA, OCT, and FA in DDME at 12 months. There seems to be no difference in our results between intravitreal bevacizumab at doses of 1.25 mg or 2.5 mg. In addition, our results suggest the need for at least three injections a year to maintain the BCVA results. PMID- 19189121 TI - Consequences of perinatal treatment with L-arginine and antioxidants for the renal transcriptome in spontaneously hypertensive rats. AB - Treating spontaneously hypertensive rats (SHR) with L-arginine, taurine, and vitamins C and E (ATCE) during nephrogenesis (2 weeks before to 4 weeks after birth) persistently lowers blood pressure. Hypothetically, differential gene expression in kidney of SHR vs. normotensive Wistar-Kyoto rats (WKY) is partially corrected by maternal ATCE in SHR. Differential gene expression in 2-days, 2 weeks, and 48-week-old rats was studied using oligonucleotide chips. Transcription factor binding sites (TFBS) of differentially expressed genes were analyzed in silico. Differential gene expression varied between SHR+ATCE and SHR, suggesting both direct and indirect effects; but, few genes were modulated toward WKY level and there was little overlap between ages. TFBS analysis suggests less Elk-1-driven gene transcription in both WKY and SHR+ATCE vs. SHR at 2 days and 2 weeks. Concluding, in SHR, persistent antihypertensive effects of maternal ATCE are not primarily due to persistent corrective transcription. Less Elk-1-driven transcription at 2 days and 2 weeks may be involved. PMID- 19189122 TI - The hemiplegic migraine-associated Y1245C mutation in CACNA1A results in a gain of channel function due to its effect on the voltage sensor and G-protein mediated inhibition. AB - Mutations in the gene encoding the pore-forming alpha(1A) subunit of P/Q Ca(2+) channels (CACNA1A) are linked to familial hemiplegic migraine. CACNA1A Y1245C is the first missense mutation described in a subject affected with childhood periodic syndromes that evolved into hemiplegic migraine. Y1245C is also the first amino acid change described in any S1 segment of CACNA1A in a hemiplegic migraine background. We found that Y1245C induced a 9-mV left shift in the current-voltage activation curve, accelerated activation kinetics, and slowed deactivation kinetics within a wide range of voltage depolarizations. Y1245C also left-shifted the voltage-dependent steady-state inactivation with a significant increase in steepness, suggesting a direct effect on the P/Q channel voltage sensor. Moreover, Y1245C reduced Gbetagamma subunits-dependent channel inhibition probably by favoring Gbetagamma dissociation from the channel; an effect also observed using action-potential-like waveforms of different durations. The formation of a new disulfide bridge between cysteines may contribute to the Y1245C effects on activation and Gbetagamma inhibition of the channel, as they were significantly reversed by the sulphydryl-reducing agent dithiothreitol. Together, our data suggest that Y1245C alters the structure of the alpha(1A) voltage sensor producing an overall gain of channel function that may explain the observed clinical phenotypes. PMID- 19189123 TI - Axial patterning of the pentaradial adult echinoderm body plan. AB - Adult echinoderms possess a highly diverged, pentaradial body plan. Developmental mechanisms underlying this body plan are completely unknown, but are critical in understanding how echinoderm pentamery evolved from bilateral ancestors. These mechanisms are difficult to study in indirect-developing species; in this study, we use the direct-developing sea urchin Heliocidaris erythrogramma, whose accelerated adult development can be perturbed by NiCl(2). We introduce a new nomenclature for the adult echinoderm axes to facilitate discussion of the radially symmetric body plan and the events required to pattern it. In sea urchins, the adult oral-aboral axis is often conflated with the long axes of the five rays; we identify these as distinct body axes, the proximodistal (PD). In addition, we define a circular axis, the circumoral (CO), along which the division into five sectors occurs. In NiCl(2)-treated larvae, aspects of normal PD pattern were retained, but CO pattern was abolished. Milder treatments resulted in relatively normal juveniles ranging from biradial to decaradial. NiCl(2) treatment had no effect either on mesodermal morphology or on the ectodermal gene expression response to an inductive mesodermal signal. This suggests that the mesoderm does not mediate the disruption of CO patterning by NiCl(2). In contrast, mesodermal signaling may explain the presence of PD pattern in treated larvae. However, variations in appendage pattern suggest that ectodermal signals are also required. We conclude that CO patterning in both germ layers is dependent on ectodermal events and PD patterning is controlled by mutual ectoderm-mesoderm signaling. PMID- 19189124 TI - Genomic characteristics of soft tissue sarcomas. AB - Studies on the molecular mechanisms behind soft tissue sarcoma development have disclosed that these malignancies are as genetically heterogeneous as they are clinically and morphologically diverse. Much of the genetic information on soft tissue sarcomas is still limited to the genomic level, as detected by chromosome banding analysis or comparative genomic hybridization. Based on the results of such studies, soft tissue sarcomas may be broadly dichotomized into one group, accounting for approximately 20% of the cases, characterized by specific balanced translocations, and one group typically showing massive chromosomal rearrangements leading to recurrent, but non-specific, structural and numerical rearrangements. As summarized in this review, the genomic characterization of soft tissue sarcomas has not only provided cell biologists with decisive information on the parts of the genome that may harbor genes that are essential for tumor development but also given the clinicians involved in the management of these patients a valuable diagnostic tool. PMID- 19189125 TI - Turquoise to dark green organs at autopsy. AB - We report the case of a 72-year-old man who died in septic shock following pancreatectomy. At autopsy, organs were discoloured with a rapid colour change from turquoise to dark green, especially of the myocardium. The patient had received 200 mg methylene blue (MB), i.v., for treatment of septic shock 90 min prior to death. Analysis of tissue samples by liquid extraction and liquid chromatography coupled to tandem mass spectrometry demonstrated different concentrations of MB and its metabolites azure A and B in the heart, lungs, kidneys, and liver. Our findings clearly demonstrate the relation of MB administration and organ discolouration at autopsy and shed a new light on MB distribution and accumulation in septic shock. PMID- 19189126 TI - Dendritic cell sarcomas/tumours of the breast: report of two cases. AB - Extranodal follicular dendritic cell sarcoma/tumours (FDCS/Ts) and interdigitating dendritic cell sarcoma/tumours (IDCS/Ts) are rare neoplasms. We present two cases of FDCS/T and IDCS/T of the breast. The FDCS/T case (case 1) presented in a 31-year-old woman and the IDCS/T case (case 2) in a 67-year-old woman who both showed a firm lump in the left breast. The FDCS/T lesion superficially appeared as an anaplastic carcinoma and the IDCS/T was reminiscent of a spindle cell sarcomatoid carcinoma. Nevertheless both lesions were negative for keratins while case 1 displayed neoplastic cells strongly positive for CD21, vimentin and focally for CD68 and S-100 protein. The tumour cells of case 2 were positive for S-100, CD68 and CD45. In breast, an unusual keratin negative tumour composed predominantly of spindle cells arranged in fascicles, storiform pattern or whorls with a lymphoid rich stroma should raise suspicion for FDCS/Ts or IDCS/Ts. The distinction from malignant tumours with similar features is discussed. PMID- 19189127 TI - Inflammatory fibroid polyp of the small bowel with a mutation in exon 12 of PDGFR alpha. AB - Inflammatory fibroid polyp (IFP) is a benign reactive uncommon submucosal lesion of the gastrointestinal tract, the small intestine being the most common site of origin. Histologically, IFPs are characterized by spindle cells, a heavy inflammatory infiltrate including eosinophils and onion-sheet-like formation of lesional cells around blood vessels. We present a case report of an IFP harboring an activation mutation in the PDGFR alpha gene. The lesion was positive for CD34, PDGFR alpha, and p-PDGFR alpha immunostaining but was negative for c-KIT and desmin. After a sequencing analysis of KIT and PDGFR alpha, a mutation consisting of an in-frame deletion of codons 567-571 and a missense mutation in codon 566 (S566R) of PDGFR alpha was observed. This mutation could activate key cellular pathways with involvement in the pathogenesis of this entity. We concluded that more studies are necessary in order to clarify if this finding is a biologically distinct behavior or, on the contrary, represents a specific feature of the IFP. PMID- 19189128 TI - Pre-hatching exposure to water mold reduces size at metamorphosis in the moor frog. AB - Developmental plasticity is increasingly recognized as important for ecological and evolutionary processes. However, few studies consider the potential for delayed effects of early environments. Here, we show that tadpoles hatching from clutches exposed to water mold (Saprolegnia) have 20% decreased mass at metamorphosis, despite no further exposure subsequent to hatching. The effects were consistent across four populations that have previously been shown to vary in their resistance to infection during embryonic development. Contrary to expectations, time to hatching or metamorphosis was not affected, suggesting that the results do not reflect an evolved escape strategy from infected waters triggered by embryonic conditions. Instead, decreased mass at metamorphosis may arise from carry-over effects of impaired embryo development. Such strong links across developmental stages have potential consequences for the evolution of plasticity and the responses of populations to emergent infections. PMID- 19189129 TI - Inferring associations among parasitic gamasid mites from census data. AB - Within a community, the abundance of any given species depends in large part on a network of direct and indirect, positive and negative interactions with other species, including shared enemies. In communities where experimental manipulations are often impossible (e.g., parasite communities), census data can be used to evaluate the strength or frequency of positive and negative associations among species. In ectoparasite communities, competitive associations can arise because of limited space or food, but facilitative associations can also exist if one species suppresses host immune defenses. In addition, positive associations among parasites could arise merely due to shared preferences for the same host, without any interaction going on. We used census data from 28 regional surveys of gamasid mites parasitic on small mammals throughout the Palaearctic, to assess how the abundance of individual mite species is influenced by the abundance and diversity of other mite species on the same host. After controlling for several confounding variables, the abundance of individual mite species was generally positively correlated with the combined abundances of all other mite species in the community. This trend was confirmed by meta-analysis of the results obtained for separate mite species. In contrast, there were generally no consistent relationships between the abundance of individual mite species and either the species richness or taxonomic diversity of the community in which they occur. These patterns were independent of mite feeding mode. Our results indicate either that synergistic facilitative interactions among mites increase the host's susceptibility to further attacks (e.g., via immunosuppression) and lead to different species all having increased abundance on the same host, or that certain characteristics make some host species preferred habitats for many parasite species. PMID- 19189130 TI - Distance decay of similarity among parasite communities of three marine invertebrate hosts. AB - The similarity in species composition between two communities generally decays as a function of increasing distance between them. Parasite communities in vertebrate definitive hosts follow this pattern but the respective relationship in intermediate invertebrate hosts of parasites with complex life cycles is unknown. In intermediate hosts, parasite communities are affected not only by the varying vagility of their definitive hosts (dispersing infective propagules) but also by the necessary coincidence of all their hosts in environmentally suitable localities. As intermediate hosts often hardly move they do not contribute to parasite dispersal. Hence, their parasite assemblages may decrease faster in similarity with increasing distance than those in highly mobile vertebrate definitive hosts. We use published field survey data to investigate distance decay of similarity in trematode communities from three prominent coastal molluscs of the Eastern North-Atlantic: the gastropods Littorina littorea and Hydrobia ulvae, and the bivalve Cerastoderma edule. We found that the similarity of trematode communities in all three hosts decayed with distance, independently of local sampling effort, and whether or not the parasites used the mollusc as first or second intermediate host in their life cycle. In H. ulvae, the halving distance (i.e. the distance that halves the similarity from its initial similarity at 1 km distance) for the trematode species using birds as definitive hosts was approximately two to three times larger than for species using fish. The initial similarities (estimated at 1 km distance) among trematode communities were relatively higher, whereas mean halving distances were lower, compared to published values for parasite communities in vertebrate hosts. We conclude that the vagility of definitive hosts accounts for a high similarity at the local scale, while the strong decay of similarity across regions is a consequence of the low probability that all necessary hosts and suitable environmental conditions coincide on a large scale. PMID- 19189131 TI - Why do parasitized hosts look different? Resolving the "chicken-egg" dilemma. AB - Phenotypic differences between infected and non-infected hosts are often assumed to be the consequence of parasite infection. However, pre-existing differences in hosts' phenotypes may promote differential susceptibility to infection. The phenotypic variability observed within the host population may therefore be a cause rather than a consequence of infection. In this study, we aimed at disentangling the causes and the consequences of parasite infection by calculating the value of a phenotypic trait (i.e., the growth rate) of the hosts both before and after infection occurred. That procedure was applied to two natural systems of host-parasite interactions. In the first system, the infection level of an ectoparasite (Tracheliastes polycolpus) decreases the growth rate of its fish host (the rostrum dace, Leuciscus leuciscus). Reciprocally, this same phenotypic trait before infection modulated the future level of host sensitivity to the direct pathogenic effect of the parasite, namely the level of fin degradation. In the second model, causes and consequences linked the growth rate of the fish host (the rainbow smelt, Osmerus mordax) and the level of endoparasite infection (Proteocephalus tetrastomus). Indeed, the host's growth rate before infection determined the number of parasites later in life, and the parasite biovolume then decreased the host's growth rate of heavily infected hosts. We demonstrated that reciprocal effects between host phenotypes and parasite infection can occur simultaneously in the wild, and that the observed variation in the host phenotype population was not necessarily a consequence of parasite infection. Disentangling the causality of host-parasite interactions should contribute substantially to evaluating the role of parasites in ecological and evolutionary processes. PMID- 19189132 TI - Which children with nephrotic syndrome benefit from cyclosporin A or cyclophosphamide? PMID- 19189133 TI - Hyporeninemic hypoaldosteronism in a child with chronic kidney disease--is this condition renoprotective? PMID- 19189134 TI - Minimal change nephrotic syndrome associated with immune dysregulation, polyendocrinopathy, enteropathy, X-linked syndrome. AB - Several studies have suggested that T cell-producing permeability factors might lead to proteinuria in minimal change nephrotic syndrome (MCNS). However, it is still unclear whether T-cell abnormalities cause MCNS. Immune dysregulation, polyendocrinopathy, enteropathy, X-linked (IPEX) syndrome is a rare disorder of the immune regulation system, which leads to severe autoimmune phenomena including autoimmune enteropathy, atopic dermatitis with high levels of serum immunoglobulin E (IgE), type 1 diabetes mellitus (T1DM), and severe infection such as sepsis, which frequently result in death within the first 2 years of life. This disease is caused by mutations in the FOXP3 gene that result in the defective development of regulatory T (Treg) cells. This report describes a 5 year-old boy with IPEX syndrome with a 3 bp deletion in the FOXP3 gene (c.748 750delAAG, p.250K.del) and a paucity of CD4(+) CD25(+) FOXP3(+) T cells. The boy's condition was complicated by MCNS in addition to many IPEX-related manifestations, such as atopic dermatitis, T1DM, enteropathy, sepsis and hemolytic anemia. This is the first report of IPEX syndrome complicated by MCNS, and our findings imply that Treg cell dysfunction may be crucial for the development of MCNS. PMID- 19189135 TI - Chylopericardial tamponade secondary to superior vena cava thrombosis in a child with nephrotic syndrome. AB - We report on a 4-year-old girl with steroid-resistant nephrotic syndrome who presented with life-threatening chylopericardial tamponade with chylothorax secondary to superior vena cava thrombosis. This is the first case report of this complication in a child. We outline the treatment, which included emergency pericardiocentesis and thoracostomy, parentral anticoagulation and dietary modifications. This therapeutic regimen resulted in a good outcome. PMID- 19189136 TI - Malignant paraganglioma associated with succinate dehydrogenase subunit B in an 8 year-old child: the age of first screening? AB - Several studies have shown that patients with succinate dehydrogenase subunit B (SDHB) mutations have a very high risk for developing malignant paragangliomas. However, there is no consensus of what age screening for paragangliomas should start. We report a case of an 8-year-old white girl with a 3-year history of catecholamine excess-related complaints who was diagnosed with a malignant SDHB associated mediastinal paraganglioma. The patient presented with intermittent sweating, headache, nausea, vomiting, fatigue and weight loss that had been present since she was 5 years of age. A large posterior mediastinal mass measuring 6.4 cm x 3.1 cm x 4.6 cm was discovered on computed tomography (CT) and magnetic resonance imaging (MRI). Laboratory data included an elevated level of urine normetanephrine of 45,400 microg/g creatinine (upper reference limit 718 microg/g) and elevated level of plasma normetanephrine of 62.4 nmol/l (upper reference limit <0.90 nmol/l). She was diagnosed with a thoracic paraganglioma and subsequently underwent surgical removal of the tumor and two lymph nodes. Histopathologic examination confirmed metastatic paraganglioma. Postoperatively, her blood pressure normalized and plasma normetanephrine levels remained normal. Our patient first presented with paraganglioma-associated signs and symptoms at the young age of 5 years. This case clearly illustrates the need for increased vigilance and screening for paragangliomas in families with SDHB at a younger age than previously thought. PMID- 19189137 TI - Non-urate transporter 1-related renal hypouricemia and acute renal failure in an Israeli-Arab family. AB - Idiopathic renal hypouricemia (IRHU) is a rare hereditary disease, predisposing the individual to exercise-induced acute renal failure (EIARF) and nephrolithiasis, and it is characterized by increased clearance of renal uric acid. Most of the described patients are Japanese, who have loss-of-function mutations in the SLC22A12 gene coding for the human urate transporter 1 (URAT1) gene. An 18-year-old youth, who was admitted for EIARF due to IRHU, and six consanguineous Israeli-Arab family members were included in the study. The family members were tested for fractional excretion of uric acid and molecular analysis of the URAT1 gene. Four family members, including the proband, had very low levels of blood uric acid and high rate of fractional excretion (FE urate> 100%) of uric acid. Genetic analysis of the affected family members did not reveal a mutation in the coding regions and intron-exon boundaries of SCL22A12. Haplotype analysis excluded SCL22A12 involvement in the pathogenesis, suggesting a different gene as a cause of the disease. We herein describe the first Israeli Arab family with IRHU. A non-URAT1 genetic defect that causes decreased reabsorption or, more probably, increased secretion of uric acid, induces IRHU. Further studies are required in order to elucidate the genetic defect. PMID- 19189138 TI - Micelles obtained by aggregation of gemini surfactants containing the CCK8 peptide and a gadolinium complex. AB - Two gemini surfactants, [C18CysL5CCK8](2) and [C18CysDTPAGlu](2), containing, respectively, the CCK8 peptide and the DTPAGlu chelating agent or its gadolinium complex have been prepared by linking lipophilic chains through a disulfide bond between two cysteine residues. The two surfactants aggregate in water solution forming pure or mixed micelles, with a critical micellar concentration in the 5 x 10(-6)-5 x 10(-5) mol kg(-1) range, as measured by fluorescence spectroscopy. As indicated by small-angle neutron scattering, the shape and size of the micelles are influenced by the temperature: increasing temperature leads to progressive reduction of the size of the supramolecular aggregates. Cylindrical structures found at lower temperatures (10-40 degrees C) evolve into ellipsoidal micelles at 50-80 degrees C. Furthermore, the surface-exposed CCK8 peptide changes its conformation above a transition temperature of approximately 45 degrees C, going from a beta-sheet to a random-coil structure, as indicated by circular dichroism measurements. The mixed aggregate obtained by coaggregation of the two gemini based amphiphilic compounds, [C18CysDTPAGlu(Gd)](2) and [C18CysL5CCK8](2) in 70:30 molar ratio, represents the first example of a peptide-containing gemini surfactant as a potential target-selective contrast agent in MRI. In fact, it presents a high relaxivity value of the gadolinium complex, 21.5 mM(-1) s(-1), and the CCK8 bioactive peptide exposed on the external surface is therefore capable of selective targeting of the cholecystokinin receptors. PMID- 19189139 TI - Low-frequency vibrational modes and infrared absorbance of red, blue and green opsin. AB - Vibrational excitations of low-frequency collective modes are essential for functionally important conformational transitions in proteins. We carried out an analysis of the low-frequency modes in the G protein coupled receptors (GPCR) family of cone opsins based on both normal-mode analysis and molecular dynamics (MD) simulations. Power spectra obtained by MD can be compared directly with normal modes. In agreement with existing experimental evidence related to transmembrane proteins, cone opsins have functionally important transitions that correspond to approximately 950 modes and are found below 80 cm(-1). This is in contrast to bacteriorhodopsin and rhodopsin, where the important low-frequency transition modes are below 50 cm(-1). We find that the density of states (DOS) profile of blue opsin in a solvent (e.g. water) has increased populations in the very lowest frequency modes (<15 cm(-1)); this is indicative of the increased thermostability of blue opsin. From our work we found that, although light absorption behaves differently in blue, green and red opsins, their low-frequency vibrational motions are similar. The similarities and differences in the domain motions of blue, red and green opsins are discussed for several representative modes. In addition, the influence of the presence of a solvent is reported and compared with vacuum spectra. We thus demonstrate that terahertz spectroscopy of low-frequency modes might be relevant for identifying those vibrational degrees of freedom that correlate to known conformational changes in opsins. PMID- 19189140 TI - Misleading initial histological diagnosis of a polymorphous low-grade adenocarcinoma in situ ex pleomorphic adenoma-a case report. AB - INTRODUCTION: Polymorphous low-grade adenocarcinoma (PLGA) are frequent tumours of palatinal minor salivary glands. They appear clinically as solid mass located beneath intact surface epithelium, thus quite similar with benign neoplasm. PLGA displays a low tendency of aggressive behaviour. The correct aetiology of this disorder is still unknown. CASE REPORT: In this contribution, a PLGA is reported which was located in a pleomorphic adenoma (PA). Out of an initially incisional biopsy, only the benign part of the lesion was diagnosed. Definitive histological examination of the whole tumour revealed a small malignant fraction of the specimen besides a major part of benign tissue formations (PA). CONCLUSION: This case shows the uncertain confidence of incisional biopsy, the variably biologic behaviour of PA, providing hints for consideration of the PLGA aetiology and highlights both the necessity to remove whole PA-like lesions as well as to perform systematically histological examination of whole specimens. PMID- 19189141 TI - Occipital condyle fractures: incidence and clinical follow-up at a level 1 trauma centre. AB - The purpose of the study was to investigate the incidence, management, and outcomes of occipital condyle fractures at a level 1 trauma center. Blunt trauma patients with occipital condyle fracture admitted to a level 1 trauma center over a 3-year period were identified. Prospective clinical and functional follow-up was undertaken, including further radiographic imaging. The incidence of occipital condyle fracture in patients presenting to our level 1 trauma center was 1.7/1,000 per year. Twenty-four patients were followed up at a mean of 27 months post-injury. There was one case of isolated occipital condyle fracture; all other patients had sustained additional orthopedic, cervical spine, and/or head injury. Seven (29%) patients sustained unilateral Type III avulsion fractures, none of which were isolated injuries. Traumatic brain injury was detected in 46% of study patients, and 42% had cervical spine injury. External halothoracic immobilization was used in 33% of cases. Fracture union with anatomical alignment occurred in 21 patients (88%). No patient had cranial nerve deficit at admission or follow-up. Three patients (12.5%) had moderate to severe neck pain/disability at follow-up, all of whom had sustained multiple injuries. Occipital condyle fractures most frequently occur in conjunction with additional injuries, particularly head and cervical spine injuries. Most cases can be managed successfully nonoperatively. Functional outcome is generally determined by pain and disability related to other injuries, rather than occipital fracture configuration. PMID- 19189142 TI - Dumbbell-shaped jugular foramen schwannomas: surgical management, outcome and complications on a series of 16 patients. AB - Schwannomas of the lower cranial nerves are very rare and the dumbbell-shaped ones are even rarer. The authors report their experience in managing such lesions, usually presenting either with intra- and/or extra-cranial extension through an enlarged jugular foramen. The juxtacondylar approach without sacrificing the labyrinth was used; clinical, radiological and complication features are discussed and analysed. This is a case series study on 16 consecutive patients with lower cranial nerves schwannomas surgically managed during a 14-year period using the juxtacondylar approach. In 13 cases, a complete resection has been achieved whereas in three, the excision was near total. The tumour nerve origin has been identified only in ten cases (62.5%). No death or additional post-operative cranial nerve deficits occurred. Aspiration pneumonia developed in one patient and cerebrospinal fluid leak in another. Pre-operative lower cranial nerve deficits improved in all patients. At a mean follow-up of 6.6 years (range 2-14 years), no radiological tumour recurrence was recorded amongst the patients having complete resection as well as no tumour progression in the group of near total removal. Jugular foramen schwannomas can be radically and safely resected with no additional neurological deficit if a careful pre operative evaluation and the appropriate surgical approach is implemented. Finally, full cranial nerve functional recovery may be expected after complete resection. PMID- 19189144 TI - Enhanced production of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) copolymer with manipulated variables and its properties. AB - Cupriavidus sp. USMAA1020, a local isolate was able to biosynthesis poly(3 hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] copolymer with various 4HB precursors as the sole carbon source. Manipulation of the culture conditions such as cell concentration, phosphate ratio and culture aeration significantly affected the synthesis of P(3HB-co-4HB) copolymer and 4HB composition. P(3HB-co 4HB) copolymer with 4HB compositions ranging from 23 to 75 mol% 4HB with various mechanical and thermal properties were successfully produced by varying the medium aeration. The physical and mechanical properties of P(3HB-co-4HB) copolymers were characterized by NMR spectroscopy, gel-permeation chromatography, tensile test, and differential scanning calorimetry. The number-average molecular weights (M (n)) of copolymers ranged from 260 x 10(3) to 590 x 10(3)Da, and the polydispersities (M (w)/M (n)) were between 1.8 and 3.0. Increases in the 4HB composition lowered the molecular weight of these copolymers. In addition, the increase in 4HB composition affected the randomness of copolymer, melting temperature (T (m)), glass transition temperature (T (g)), tensile strength, and elongation to break. Enzymatic degradation of P(3HB-co-4HB) films with an extracellular depolymerase from Ochrobactrum sp. DP5 showed that the degradation rate increased proportionally with time as the 4HB fraction increased from 17 to 50 mol% but were much lower with higher 4HB fraction. Degradation of P(3HB-co 4HB) films with lipase from Chromobacterium viscosum exhibited highest degradation rate at 75 mol% 4HB. The biocompatibility of P(3HB-co-4HB) copolymers were evaluated and these copolymers have been shown to support the growth and proliferation of fibroblast cells. PMID- 19189143 TI - Isolation and characterization of cellulose-degrading bacteria from the deep subsurface of the Homestake gold mine, Lead, South Dakota, USA. AB - The present study investigated the cultivable mesophilic (37 degrees C) and thermophilic (60 degrees C) cellulose-degrading bacterial diversity in a weathered soil-like sample collected from the deep subsurface (1.5 km depth) of the Homestake gold mine in Lead, South Dakota, USA. Chemical characterization of the sample by X-ray fluorescence spectroscopy revealed a high amount of toxic heavy metals such as Cu, Cr, Pb, Ni, and Zn. Molecular community structures were determined by phylogenetic analysis of 16S rRNA gene sequences retrieved from enrichment cultures growing in presence of microcrystalline cellulose as the sole source of carbon. All phylotypes retrieved from enrichment cultures were affiliated to Firmicutes. Cellulose-degrading mesophilic and thermophilic pure cultures belonging to the genera Brevibacillus, Paenibacillus, Bacillus, and Geobacillus were isolated from enrichment cultures, and selected cultures were studied for enzyme activities. For a mesophilic isolate (DUSELG12), the optimum pH and temperature for carboxymethyl cellulase (CMCase) were 5.5 and 55 degrees C, while for a thermophilic isolate (DUSELR7) they were 5.0 and 75 degrees C, respectively. Furthermore, DUSELG12 retained about 40% CMCase activity after incubation at 60 degrees C for 8 h. Most remarkably, thermophilic isolate, DUSELR7 retained 26% CMCase activity at 60 degrees C up to a period of 300 h. Overall, the present work revealed the presence of different cellulose-degrading bacterial lineages in the unique deep subsurface environment of the mine. The results also have strong implications for biological conversion of cellulosic agricultural and forestry wastes to commodity chemicals including sugars. PMID- 19189146 TI - Determination of the influence of stent strut thickness using the finite element method: implications for vascular injury and in-stent restenosis. AB - Many clinical studies, including the ISAR-STEREO trial, have identified stent strut thickness as an independent predictor of in-stent restenosis where thinner struts result in lower restenosis than thicker struts. The aim of this study was to more conclusively identify the mechanical stimulus for in-stent restenosis using results from such clinical trials as the ISAR-STEREO trial. The mechanical environment in arteries stented with thin and thicker strut stents was investigated using numerical modelling techniques. Finite element models of the stents used in the ISAR-STEREO clinical trial were developed and the stents were deployed in idealized stenosed vessel geometries in order to compare the mechanical environment of the vessel for each stent. The stresses induced within the stented vessels by these stents were compared to determine the level of vascular injury caused to the artery by the stents with different strut thickness. The study found that when both stents were expanded to achieve the same initial maximum stent diameter that the thinner strut stent recoiled to a greater extent resulting in lower luminal gain but also lower stresses in the vessel wall, which is hypothesised to be responsible for the lower restenosis outcome. This study supports the hypothesis that arteries develop restenosis in response to injury, where high vessel stresses are a good measure of that injury. This study points to a critical stress level in arteries, above which an aggressive healing response leads to in-stent restenosis in stented vessels. Stents can be designed to reduce stresses in this range in arteries using preclinical tools such as numerical modelling. PMID- 19189145 TI - A new challenge-development of test systems for the infochemical effect. AB - BACKGROUND, AIM, AND SCOPE: Many-if not all-organisms depend on so-called infochemicals, chemical substances in their surroundings which inform the receivers about their biotic and abiotic environment and which allow them to react adequately to these signals. Anthropogenic substances can interfere with this complex chemical communication system. This finding is called infochemical effect. So far, it is not known to what extent anthropogenic discharges act as infochemicals and influence life and reproduction of organisms in the environment because adequate testing methods to identify chemicals which show the infochemical effect and to quantify their effects have not been developed yet. The purpose of this article is to help and find suitable test designs. MAIN FEATURES: Test systems used in basic research to elucidate the olfactory cascade and the communication of environmental organisms by infochemicals are plentiful. Some of them might be the basis for a quantified ecotoxicological analysis of the infochemical effect. In principle, test systems for the infochemical effect could be developed at each step of the chemosensory signal transduction and processing cascade. RESULTS: Experimental set-ups were compiled systematically under the aspect whether they might be usable for testing the infochemical effect of single chemicals in standardized quantifying laboratory experiments. For an appropriate ecotoxicological assessment of the infochemical effect, experimental studies of many disciplines, such as molecular biology, neurobiology, physiology, chemical ecology, and population dynamics, should be evaluated in detail before a decision can be made which test system, respectively which test battery, might be suited best. The test systems presented here are based on the knowledge of the genetic sequences for olfactory receptors, binding studies of odorants, signal transmission, and reactions of the receivers on the level of the organisms or the populations. The following basic approaches are conceivable to identify the role of an infochemical: binding studies to the odorant-binding protein or to the odorant receptor binding protein (e.g., by in situ hybridization and immunohistochemical studies), measurement of electrical signals of the receptor cells in the tissue (e.g., electroolfactograms, electroantennograms), registration of phenotypic changes (e.g., observation under the microscope), behavioral tests (e.g., in situ online biomonitoring, use of T-shaped olfactometers, tests of avoidance responses), measurement of population changes (e.g., cell density or turbidity measurements), and multispecies tests with observation of community structure and community function. The main focus of this study is on aquatic organisms. DISCUSSION: It is evident that the infochemical effect is a very complex sublethal endpoint, and it needs further studies with standardized quantitative methods to elucidate whether and to what extent the ecosystem is affected. The collection of approaches presented here is far from being complete but should serve as a point of depart for further experimental research. CONCLUSIONS: This article is the first to compare various approaches for testing the infochemical effect. The development of a suitable test system will not be easy as there are a multitude of relevant chemicals, a multitude of relevant receptors, and a multitude of relevant reactions, and it must be expected that the effective concentrations are very low. The chemical communication is of utmost importance for the ecosystem and justifies great endeavors to find solutions to these technical problems. RECOMMENDATIONS AND PERSPECTIVES: The infochemical effect is a new chapter in ecotoxicology. Will a new endpoint, the so-called infochemical effect, be required in addition to the actual standard test battery of Annex 5 to Commission Directive 92/69/EEC (EC 1992)? Finding the answer to this question is a big challenge that could be met by a comprehensive research project. PMID- 19189148 TI - [Self-help groups in healthcare]. PMID- 19189147 TI - Wave intensity analysis and the development of the reservoir-wave approach. AB - The parameters of wave intensity analysis are calculated from incremental changes in pressure and velocity. While it is clear that forward- and backward-traveling waves induce incremental changes in pressure, not all incremental changes in pressure are due to waves; changes in pressure may also be due to changes in the volume of a compliant structure. When the left ventricular ejects blood rapidly into the aorta, aortic pressure increases, in part, because of the increase in aortic volume: aortic inflow is momentarily greater than aortic outflow. Therefore, to properly quantify the effects of forward or backward waves on arterial pressure and velocity (flow), the component of the incremental change in arterial pressure that is due only to this increase in arterial volume--and not, fundamentally, due to waves--first must be excluded. This component is the pressure generated by the filling and emptying of the reservoir, Otto Frank's Windkessel. PMID- 19189149 TI - [Evolution of health-related self-help. Experience from 40 years of work in self help]. AB - The characterization of health-related self-help often includes social demands, which indicate that the knowledge on this healthrelated self-help is very small. On the basis of 40 years experience in self-help, the article traces different lines of evolution in health-related self-help. The article describes the development of self-help structures and their success. It illustrates the association between self-help and self-help organisations. It demonstrates the social legitimation of work in self-help. Finally, the article describes current challenges for the work in health-related self-help. Based on the evidence of theses new challenges, for example, the increased significance of rare diseases, the growing scientific foundation of health care or the discussions on the different offers for health-related information and consultation, the article demonstrates the future trends in health-related self-help. PMID- 19189150 TI - [Development, situation and perspective of self-help support in Germany]. AB - Self-help groups and self-help associations are an important part of the social security system. In Germany, self-help contact points, senior citizen centers, volunteer agencies, citizen centers and multi-generation houses combine citizen participation with innovative professional services. Unfortunately, there is no guarantee of continuous financial support for these important, locally administered institutions. There are about 280 self-help contact points and more than 400 federal self-help associations that support and promote self-help in Germany. Healthy communities, healthy workplaces and healthy people need a decentralized system of self-help programs operated at local and regional levels, in districts and towns. Thereby, professional support systems that operate self help programs and promote citizen participation in the self-help programs must be managed in a similar regional format. New forms of cooperation from the regional and local governments, private companies, and citizen engagement already exist. Additionally, regional projects of integrated maintenance systems with the regional health maintenance institutions have been established. Currently, the central challenges of the self-help programs are quality development, inclusion of people with social disadvantages and of people with migrational background. The essential prerequisites for this work are continuous financial support and a politically supported infrastructure, which is in fact an important health investment. PMID- 19189151 TI - [Patient self-help, user perspective and health services research]. AB - Patient's individual responsibility, self-help and active co-operation are essential for the health care system. This paper presents a synopsis of research in self-help groups and self-help organizations. Studies provide evidence that self-help initiatives are an important contribution to population health. Especially self-help groups increase mutual aid and social support, knowledge about the disease and its consequences and possibilities for changing attitudes of the group members and their social environment. They disburden primary networks and families and reinforce an 'intelligent' utilization of professional health care services. There has been as yet hardly any systematic investigation of the activities of self-help organizations, except from counseling services. Self-help groups and organizations increasingly receive financial support from the social security system and have increasingly become part of the health care system. However, the co-operation between self-help initiatives and the professional health care system is characterized by an imbalance between laymen and experts. Given these results, self-help, user perspective and the health related behavior of laymen should obtain more significance in the conceptualization and definitions of health services research. PMID- 19189152 TI - [Cooperation of physicians and self-help groups. Scientific findings]. AB - Chronic diseases and handicaps are widespread among the population. Considering the psycho-social components of many diseases, medical therapy alone is often not sufficient. Self-help groups meet emotional or other non-medical needs and support the coping process of their members. A widespread assumption is that cooperation between physicians and self-help groups promotes the quality of medical care and quality of life of the afflicted. Evidently, physicians and self help groups can complement each other. This article presents scientific findings about mutual contacts and cooperation of office-based physicians, hospitals, rehabilitation clinics, on the one hand, and self-help groups, on the other. A variety of modes of cooperation are described as well as mutual perceptions, expectations, the perceived quality of cooperation and cooperation-limiting factors. PMID- 19189153 TI - [Cooperation between physicians and self-help groups in the system of ASHIP. Practical experience]. AB - In recent years, there has been increased cooperation between doctors' associations and patient self-help groups. Much of this cooperation is dependent on the engagement and involvement of individuals. However, a far reaching, established type of cooperation is still lacking. Several studies show that there are many advantages to be gained from working with self-help organizations. In Germany, ambulatory medical care is managed by the Association of Statutory Health Insurance Physicians (ASHIP). In order to develop the cooperation between patient self-help groups and ASHIP physicians, several information centers (KOSA) for these groups have been established. These information centers support direct communication between patient self-help groups and physicians by means of selective measures, specialist events and expert forums. A cooperation handbook is available for doctors and psychotherapists detailing case studies of the experiences of KOSA members. In addition to these information centers, cooperation is supported by patient advisory boards, round table discussions and symposia. PMID- 19189154 TI - [Integration of self-help associations into the quality management of outpatient and inpatient health care]. AB - The idea of introducing a special label for "self-help-friendly" institutions was first developed for hospitals. A demonstration project (BKK BV, the German Federal Association of Company Health Insurance Funds) was launched and organized by the local contact point for self-help groups in Hamburg. Shortly thereafter, a group of experienced self-help supporters started to implement self-help friendliness in quality management systems for ambulatory care. These endeavours, presented in this article, prove that not only professional but also user-based quality standards are becoming more and more common in quality development processes of health care services. The general legislative and political conditions for these developments have considerably improved over the past few years. This will be the foundation for a new quality of the doctor-patient relationship. PMID- 19189155 TI - [Health-related self-help of immigrants in Germany. Chances, barriers, potential]. AB - Migrants in Germany participate in health-related self-help groups nearly as frequently as native-born Germans--as long as they have good German language skills and are widely integrated into the main society. However, the situation is different for those migrants who are not or only slightly integrated. Mother tongue self-help groups of ethnic minorities would offer good chances for health related self-management and coping; however, these groups are rare. This can partly be explained by the lack of knowledge of many migrants about support possibilities for self-help groups, also because the term "self-help" has no equivalent in many languages. On the other hand, there are many barriers for self help activities: false shame attributed to certain health problems, fear of de anonymization in ethnic communities, different concepts of health and illness, a lack of judging the potential of one's own competencies and self-efficacy, and particularly illiteracy. This article describes the relevance of self-help among migrants with special regard to self-organized migrant networks and outlines the current state of discussion within the self-help stakeholders. Finally, potential multipliers, networking examples and qualification measures for promotion of health-related self-help among migrants are presented. PMID- 19189156 TI - [Performance of self-help groups and their economic evaluation]. AB - Hoffmann von Fallersleben is quoted with the sentence "Self-help is worthwhile, because it does not demand anything from others". This sounds catchy; it is, however, wrong: Self-help groups ask for support, particularly for financial resources for the work of either individual, highly organized self-help associations or for general support of self-help groups via local contact and information centers ("contact points for self-help groups"). With this request for economic "investments" in self-help, the question arises whether this is profitable for the country, the local authority or the social health insurance. In principle, the initial answer to this is: yes, the work of self-help groups is worthwhile for a single person, but also for the larger community, as various kinds of services are provided by self-help groups and organizations. Despite many surveys of members or co-operation partners which show positive effects of self-help groups, the question remains whether services of self-help groups can be measured and economically evaluated. The socio- political question regarding funding is closely connected to the idea of an economic evaluation of self-help groups. The aim of this article is to summarize and discuss which empiric approaches and findings are available on this subject. The monetary value for the work done per member of self-help groups and year lies between approximately 700 and 900 EUR. PMID- 19189157 TI - [Self-help groups conflicts of interest through sponsoring by the pharmaceutical industry]. AB - Some patient and self-help groups accept financial support from the pharmaceutical industry and medical device manufacturers. For the industry, this constitutes an increasingly important product marketing component. The acceptance of material or other support triggers psychological mechanisms which endanger objective judgement without the persons involved realizing it. Thus, patient groups may evaluate drugs or devices in a positively distorted way. PMID- 19189158 TI - [Evidence-based patient information: the example of immunotherapy for patients with multiple sclerosis]. AB - The article elucidates consideration of scientific criteria for the development and design of evidence-based patient information (EBPI). Immunotherapy of multiple sclerosis serves as an example. Since in EBPI lack of evidence or ambiguities in available evidence are explicitly communicated, processing of EBPI does not necessarily lead to certainty about benefit and harms of medical interventions. However, only if the information is comprehensive in this respect can the EBPI be regarded as a robust basis for an informed choice. EBPI requires substantial developmental efforts. Regarding the growing number of medical interventions and the half-life of information, the question of responsibility for provision of EBPI is crucial. A vision is drafted in which EBPI is driven by demand of the patients and the public and is provided according to a costs-by cause principle by those who distribute usual information hitherto. Trained patient advocates can appraise quality of information by use of instruments that consider criteria of EBPI. Critical health literacy should evolve early in school and can later on enhance usefulness of EBPI for people concerned with health issues. PMID- 19189159 TI - [The principles of shared decision-making and the contribution of self-help institutions]. AB - Patient participation in decision-making has become increasingly important. Shared decision-making (SDM) describes a procedure for the physician-patient encounter and can be enhanced by training physicians, decision support tools and training patients. Results of a research consortium on SDM funded by the ministry of health showed positive effects of shared decision-making for both the patient and the physician. Self-help organizations provide information for patients and support their sovereignty dealing with their illness by counseling and training. Thus, self-help institutions already contribute to the enhancement of patient competencies for shared decision-making. A closer cooperation between research and self-help institutions can enhance the implementation of shared decision making. Possible steps are the development of closer collaborations, exchange of experiences and the common development and implementation of decision support tools. PMID- 19189160 TI - [Tasks and experiences of the Federal Government Commissioner for Patients' Affairs]. AB - On the basis of Section 140h of Social Code Book V (German abbreviation: SGB V), which had been newly included in 2004, the Federal Government Commissioner for Patients' Affairs speaks out in favor of a meticulous implementation of patients' right to education, information and joint decision-making in all spheres of the health care system, whenever the commissioner attends the respective panels or events, and also in public. Currently, the commissioner is member of a parliamentary working group which is charged with the elaboration of the contents of a law relating to patients' rights, and experts and patients' organizations are involved in this procedure as well. For this purpose, but also with regard to other key panels of the health care system, self-help organizations are important partners for collaboration, due to their commitment, their experiences and the inspirations they provide. Therefore, they have been strengthened in essential aspects by the policy of the Federal Government over the past few years. PMID- 19189161 TI - [Participation of representatives for patients in the Federal Joint Committee. Results of qualitative interviews]. AB - The GMG (Law Modernizing the German Health Care System) was introduced in 2004. It resulted in considerable changes concerning the possibility for patients to participate in decisions in the German health care system. For the first time representatives of patients in the restructured Federal Joint Committee (G-BA), elected by patients and patient-organizations, were institutionalized by law. In later legislation, this route was further pursued: GKV-WSG and the VAndG of 2007. A previous quantitative study performed by the Institute for Health Economics and Clinical Epidemiology of the University of Cologne in 2006 provided some information on the degree of the involvement of patient representatives in the decision-making process of the G-BA. The present study is an attempt to extend this information by further in-depth interviews. The qualitative interviews of patient representatives in the G-BA unravelled a basic need for improvement of their financial and structural resources. This would lead to consequences in nearly all areas of patient participation--beginning with the quality of the patient representatives' contributions, the role within the G-BA and subsequently their role in public relations. It can be concluded that health politicians can be influenced only gradually in a process of permanent discussions on these issues. PMID- 19189162 TI - [Patients' participation in the federal joint committee of the German health care system]. AB - The first step of user participation which was established by the reform of the German health care system in 2004 currently presents one of the most essential innovations in health policy. The participation of patients' and consumers' representatives in the corporate bodies of the federal joint committee ("Gemeinsamer Bundesausschuss") is, in addition to other activities, an essential contribution to more transparency in the decision process. Due to the participation and the consenting or even critical votes of the patients' representatives, the partially controversial resolutions receive more credibility and are better accepted. On the other hand, the consumers' and patients' organizations have proved to be a competent and reliable partner in order to argue for or against controversial decisions. Patients' participation reduces the legitimation deficits of the self-administrational bodies and thus helps to maintain the German health corporatism. The German health care system also benefits from patients' participation because it corresponds to the internal logic of its crucial principles of solidarity and reimbursement. PMID- 19189163 TI - [Practice, problems and perspectives of opioid substitution treatment (OST) in Germany]. AB - Opioid dependency is a complex and chronically relapsing disease with high risks of morbidity and mortality. Frequent relapses and in most of the cases a long process of maturing out characterize this disease. Opioid substitution programs with methadone, buprenorphine and other opioids are a suitable intervention and form the first choice in the treatment of this disease. In Germany, compared to its neighboring countries, this treatment was introduced relatively late. However, in the last five years, the number of patients in substitution treatment has increased significantly to more than 70,000 patients, which marks an increase of 50%, meaning that one third to one half of the estimated opiate users are being reached. Despite the widely acknowledged success with respect to the improvement of the quality of life, survival rates, and accessibility of the target groups for ongoing treatments of drug-related diseases (such as HIV/HCV infections), opioid substitution treatment is still discussed controversially. In this contribution, problems in the areas of service provision, juristic, social, health and research policies are discussed and possibilities of increasing the access and quality of this treatment are introduced. PMID- 19189165 TI - Estimated prevalence and patterns of presumed osteoporosis among older Americans based on Medicare data. AB - INTRODUCTION: Estimates of osteoporosis (OP) prevalence based on bone mineral density testing and fracture occurrence may be imprecise for small demographic groups. Medicare data are a useful supplemental source of information on OP. METHODS: We studied people ages > or = 65 years covered by Medicare 2005. Cases of presumed OP were beneficiaries with physician services or inpatient claims for OP or for an associated fracture (hip, distal forearm, spine) in 1999-2005. RESULTS: Among 911,327 beneficiaries with 6 or 7 years of Medicare coverage, the overall prevalence of OP and associated fractures was 29.7%. Prevalence was four times higher for women than men, increased with age, and was two times higher for whites, Hispanic Americans, and Asian Americans than African Americans. Among people with OP-associated fracture claims, the proportion with an OP diagnosis was 49.7% overall (women, 57.1%; men, 21.9%) and was lower for men than women and for African Americans than other ethnic groups. CONCLUSIONS: The low proportion of beneficiaries who had an OP-associated fracture and also had an OP diagnosis, particularly among men and African American women, suggests suboptimal recognition and management of OP. Study limitations included lack of validation of our definition of OP and potential misclassification of race/ethnicity. PMID- 19189166 TI - Successful treatment of pyoderma gangrenosum associated with juvenile idiopathic arthritis with a combination of topical tacrolimus and oral prednisolone. PMID- 19189167 TI - A rare case of minimal deviation adenocarcinoma of the uterine cervix in a renal transplant recipient. AB - INTRODUCTION: We report the first described case of minimal deviation adenocarcinoma of the uterine cervix in the setting of a female renal cadaveric transplant recipient. MATERIALS AND METHODS: A retrospective review of this clinical case was performed. CONCLUSION: This rare cancer represents only about 1% of all cervical adenocarcinoma. PMID- 19189169 TI - Ultrasound-assisted lipoplasty: the ad hoc situation. PMID- 19189168 TI - Systematic review and metaanalysis of statins for heterozygous familial hypercholesterolemia in children: evaluation of cholesterol changes and side effects. AB - Heterozygous familial hypercholesterolemia (heFH) affects 1 in 500 individuals. Evidence supports the low-density lipoprotein (LDL)-lowering effect of statins for adults with heFH. However, there are concerns regarding the treatment children with heFH. By performing a systematic review and metaanalysis of the published literature, this study aimed to evaluate the efficacy and safety of statins used for children with heFH. A systematic review was performed by searching multiple medical databases and citations to identify reports of randomized controlled trials of statins used to treat children with heFH. The trials were retrieved, reviewed, and subjected to metaanalysis. The search yielded 2,174 titles. Of the 63 studies retrieved and reviewed, 56 were excluded, 7 were included in the systematic review, and 4 were included in the metaanalysis. Significant heterogeneity was detected. The metaanalysis showed significant LDL lowering, high-density lipoprotein (HDL) cholesterol elevation, and increases in height and weight with statins. The metaanalysis could not be performed for many side effects of statins, but individual trials showed no significant side effects. Quality assessment showed methodologic concerns, with potential for bias. For example, six trials analyzed statin effects without intention to treat despite such a stated intention. Metaanalysis shows significant LDL lowering with statin treatment. Further studies, including epidemiologic and multicenter studies, are required. PMID- 19189170 TI - Potential effects of direct transfer payments on farmland habitat diversity in a marginal European landscape. AB - Farmland habitat diversity in marginal European landscapes changed significantly in the past decades. Further changes toward homogenization are expected, particularly in the course of European agricultural policy. Based on three alternative transfer payment schemes, we modeled spatially explicit potential effects on the farmland habitat diversity in a marginal European landscape. We defined (1) a scenario with direct transfer payments coupled to production, (2) a scenario with direct transfer payments decoupled from production, and (3) a scenario phasing out all direct transfer payments. We characterized habitat diversity with three indices: habitat richness, evenness, and rarity. The habitat pattern in 1995 served as reference for comparison. All scenarios predicted a general trend of homogenization of the farmland habitat pattern, yet to a differing extent. Transfer payments coupled to production (Scenario 1) favored the abandonment of agricultural production, particularly in low-productive areas and arable land use in more productive areas. Habitat richness and habitat evenness had intermediate values in this scenario. Decoupling transfer payments from production (Scenario 2) supported grassland as most profitable farming system. This led to a grassland-dominated landscape with low values of all habitat diversity indices. Phasing out transfer payments (Scenario 3) resulted in complete abandonment or afforestation of agricultural land and extremely low values in all habitat diversity indices. Scenario results indicate that transfer payments may prevent cessation of agricultural production, but may not counteract homogenization in marginal landscapes. Conserving high farmland habitat diversity in such landscapes may require support schemes, e.g., Pillar Two of EU Common Agricultural Policy. PMID- 19189171 TI - Sediment quality assessment in the Gulf of Gdansk (Baltic Sea) using complementary lines of evidence. AB - Sediments from Polish coastal environments were classified by a quality assessment approach that took into account trace metal and organic micropollutant concentrations, grain-size distribution, and organic carbon content. Generally, no benthic organisms were found at sites where sediments were classified as heavily polluted. However, areas characterized by a moderate contamination showed a variable composition of the benthic community and changing bioaccumulation patterns; therefore, no single species found in the Gulf of Gdansk could be considered representative of the whole benthic environment. Although sediment monitoring must be considered a suitable tool to detect hot-spot pollution areas in coastal and inland waters, it should be complemented by bioaccumulation measurements to evaluate the actual risk posed by contaminants to benthic organisms. This "biological information" allows a better appreciation of the real benthic infaunal community exposure to chemicals and can usefully complement the existing sediment quality guidelines. PMID- 19189172 TI - The importance of supratidal habitats for wintering shorebirds and the potential impacts of shrimp aquaculture. AB - Intensive black tiger shrimp (Penaeus monodon) aquaculture ponds have replaced significant areas of coastal wetlands throughout tropical Asia. Few studies have assessed potential impacts on avian foraging habitats. At Khao Sam Roi Yod National Park, Thailand, seminatural wetlands have been converted to either shrimp ponds or to salinization ponds that provide saline water for shrimp aquaculture. Although shorebirds cannot feed in aquaculture ponds, hypersaline ponds can provide productive foraging areas. Thus, the overall impact of the shrimp industry on shorebirds depends partly on the relative quality of the salt ponds compared to seminatural wetlands. In this study, we examined wintering shorebird use of tidal (N = 5 sites) and supratidal areas (four wetland sites, four salt pond sites) and compared the shorebird community (14 species), prey availability, profitability, and disturbance rates between wetlands and salt ponds. Two shorebird species fed in higher densities in wetlands, whereas seven species were more abundant in salt ponds. Large juvenile fish and dragonfly larvae were more abundant in wetlands, whereas there were more small Chironomid midge and fly larvae in salt ponds. We conclude that salt ponds might provide higher-quality foraging habitats compared to wetlands for small shorebirds species because of the abundance of small larvae. However, the shrimp aquaculture industry reduces habitat availability for shorebirds feeding on larger prey. This study demonstrates a comprehensive, multispecies approach to assess the impacts of a large-scale change in coastal habitats for wintering shorebirds. PMID- 19189173 TI - Lateral subfascial endoscopic perforating vein surgery as treatment for lateral perforating vein incompetence and venous ulceration. AB - BACKGROUND: The aim of this study was to evaluate the clinical results of lateral subfascial endoscopic perforating vein surgery (SEPS). METHODS: Our study was conducted from February 2002 to January 2007. Sixty-three patients with lateral incompetent perforating veins (IPVs) and venous ulceration were allocated to two groups. Group 1 comprised 31 patients (33 limbs) who underwent only routine surgery (saphenofemoral or saphenopopliteal ligation, stripping, phlebectomies, endovascular laser treatment [EVLT], medial SEPS, and skin grafting). Group 2 comprised 32 patients (35 limbs) who underwent our routine surgery with the addition of lateral SEPS. Bidirectional ultrasonography was performed before the operation, and 6 weeks, 6 months, and 1 year after surgery. Clinical score and disability score were collected at the same time. RESULTS: Twenty-nine (87.9%) of 33 limbs with active ulcers in group 1 healed, with recurrence in four (12.1%) limbs at follow-up at 25.9 +/- 10.0 months. All 35 limbs in group 2 healed, with recurrence in 0 (0.0%) limbs at follow-up at 25.4 +/- 10.1 months. Lateral SEPS could reduce the rate of recurrence of ulcers (Kaplan-Meier analysis). Patients in group 2 also showed improvement in clinical and disability scores, although they were not statistically significant differences (t test, p > 0.05). A significantly higher proportion of patients in group 1 had IPVs on the lateral aspect of the lower leg on ultrosonography imaging. CONCLUSIONS: Lateral SEPS could contribute to ulcer healing and reduce ulcer recurrence. The addition of lateral SEPS to routine surgery did reduce the number of IPVs. Lateral SEPS may be safe and effective for lateral perforating vein incompetence and venous ulceration. PMID- 19189175 TI - Management of upper extremity vascular injury: outcome related to the Mangled Extremity Severity Score. AB - BACKGROUND: The Mangled Extremity Severity Score (MESS) is an objective criterion for amputation prediction after lower extremity injury as well as for amputation prediction after upper extremity injury. A MESS of >or=7 has been utilized as a cutoff point for amputation prediction. In this study, we examined the result of upper extremity vascular injurty (UEVI) management in terms of the amputation rate as related to the MESS. METHODS: During January 2002 to July 2007, we reviewed patients with UEVIs at our institution. Data collections included demographic data, mechanism of injuries, injury severity score (ISS), ischemic time, MESS, pathology of UEVI, operative management, and amputation rate. Decisions to amputate the injured limbs at our institution were made individually by clinically assessing limb viability (i.e., color and capillary refill of skin; color, consistency, and contractility of muscles) regardless of the MESS. The outcome was analyzed in terms of the amputation rate related to the MESS. RESULTS: There were 52 patients with UEVIs in this study: 25 (48%) suffered blunt injuries and 27 (52%) suffered penetrating injuries. The age ranged from 15 to 59 years (mean 28.7 years). The mean ischemia time was 10.07 h. The mean ISS was 17.52. There were 12 patients (23%) with subclavian artery injuries, 3 patients (5.76%) with axillary artery injuries, 18 patients (34.61%) with brachial artery injuries, and 19 patients (36.54%) with radial artery and/or ulnar artery injuries. Primary repairs were performed in 45 patients (86.54%), with ligations in 3 patients (5.77%). An endovascular stent-graft was used in one patient (1.92%). Primary amputations were performed in three patients (5.77%). Secondary amputations (amputation after primary operation) were done in 4 of 49 patients (secondary amputation rate 8.16%). All amputation patients suffered blunt injuries and had a MESS of >or=7 (range 7-11). The overall amputation rate in this study was 13.46% (7/52 patients). Multivariate analysis revealed that the only factor significantly associated with amputation was the MESS. There were no amputations in 33 patients who had a MESS of <7. We could avoid amputation in 12 of 19 patients who had a MESS>or=7. There were no mortalities among 52 UEVI patients. CONCLUSIONS: MESS, an outcome score used to grade the severity of extremity injuries, correlates well with the risk of amputation. Nevertheless, a MESS of >or=7 does not always mandate amputation. On the other hand, the MESS is a better predictor for patients who do not require amputation when the score is <7. The decisions to amputate in patients should be made individually based on clinical signs and an intraoperative finding of irreversible limb ischemia. PMID- 19189174 TI - Postoperative fatigue: a review. AB - BACKGROUND: Postoperative fatigue (POF) significantly impacts well-being after major surgery. However, this topic has received little emphasis. METHODS: We conducted a comprehensive search on major databases with a focus on studies relevant to assessment and etiology of POF. RESULTS: POF has been measured by a number of different and inadequate instruments. It has a complicated etiology, with a number of biological and psychological factors implicated. However, the etiology of this condition has not been fully explained. CONCLUSIONS: The role of local inflammation in the development of POF requires further research. Multimodal interventions should be conducted with a focus on addressing various factors that contribute to the development and progression of POF. PMID- 19189177 TI - How to treat esophageal perforations when determinants and predictors of mortality are considered. AB - BACKGROUND: Published lethality rates of esophageal perforation (EP) vary depending on patient- and disease-related factors. This study was designed to evaluate how these factors impact death. Furthermore, we calculated the predictive accuracy of the Mortality Prediction Model (MPM II) and the Simplified Acute Physiology Score (SAPS II) for in-hospital death. Conclusions about treatment decisions were drawn based on our data and analysis of recent literature. METHODS: Every patient who was treated for EP at our department from December 2001 to July 2008 is included in this study. Logistic regression analyses of various risk factors, such as etiology, time interval, size, comorbidities, localization, type of treatment, and preexisting pathologies of the esophagus on death, were performed. RESULTS: Of the 41 patients diagnosed with EP, nine died (21%). The most important risk factor concerning death was cirrhosis of the liver (0 vs. 89% mortality; odds ratio, 208; P<0.001). Accuracy for lethality risk prediction was calculated with MPM II and SAPS II on admission, and afterward the characteristic increase that occurred was evaluated by using receiver operator characteristic curves. Optimal results were achieved by using a characteristic SAPS II increase (AUC 0.86; P: 0.009) after the patient was admitted to the intensive care unit. CONCLUSIONS: Our study was the first to demonstrate that a rapid or continuous increase more than 40 of the daily SAPS II clearly indicates that a high risk of death is imminent. This should be used as a reevaluation factor when choosing a treatment strategy. PMID- 19189178 TI - Identification and characterization of a chitinase-produced bacillus showing significant antifungal activity. AB - Strain MY75 is a gram-positive, aerobic, endospore-forming bacterium that can secrete high levels of extracellular chitinase (4.645 U/ml) when chitin powder exists as an inducer. This strain was identified as Bacillus licheniformis using the Biolog MicroLog microbial identification system and sequence analysis of 16S rDNA, gyrA and rpoB genes. Strain MY75 has the ability to inhibit the growth of Gibberella saubinetii and Aspergillus niger, two major pathogenic fungi in agriculture, and to restrain their spore germination completely. The chitinase was proved to play an important role in the strain's antifungal activity. PMID- 19189179 TI - Binding of IgA by Mycoplasma penetrans. AB - The current study shows that Mycoplasma penetrans strain GTU binds human serum immunoglobulin A (IgA) and secretory IgA but not IgG. Binding of IgA was associated almost exclusively with the lipoprotein fraction obtained by Triton X 114 fractionation of isolated M. penetrans membranes. Western immunoblot analysis of isolated membranes of M. penetrans strain GTU and of the Triton X-114 fraction showed that the major IgA-binding component was a lipoprotein with a molecular mass of 38 kDa, one of the major lipoproteins of this organism. The authors suggest that the high IgA-binding capacity of M. penetrans strain GTU may serve as a defense mechanism, conferring on this microorganism the ability to evade clearance mediated by specific IgA antibodies. PMID- 19189180 TI - Reduction of selenite to elemental red selenium by Pseudomonas sp. Strain CA5. AB - A Pseudomonas sp. that may be useful in bioremediation projects was isolated from soil. The strain is of potential value because it reduces selenite to elemental red selenium and is unusual in that it was resistant to high concentrations of both selenate and selenite. Exposure of the strain to 50, 100, and 150 mM selenite reduced growth by 28, 57, and 66%, respectively, while no change in growth was observed when the strain was exposed to 64 mM selenate, the highest level tested. Cells of the strain removed 1.7 mM selenite from the culture fluid during a 7-day incubation. A selenite reductase with a molecular weight of ~115 kD was detected in cell-free extracts and a protein with a molecular weight of ~700 kD was detected that reduced both selenate and nitrate. The bacterial isolate is a strict aerobe, reducing selenite to elemental red selenium under aerobic conditions only. Pseudomonas sp. strain CA5 might be useful as an inoculum for bioreactors used to harvest selenium from selenite-containing groundwater. 16S rRNA gene sequence alignment and fatty acid analysis were used to identify the bacterium as a novel species of Pseudomonas related to P. argentinensis, P. flavescens, and P. straminea. PMID- 19189181 TI - Thiosulfate oxidation by Comamonas sp. S23 isolated from a sulfur spring. AB - A bacterial isolate S23 capable of oxidizing thiosulfate was isolated from a sulfur spring. Strain S23 is gram-negative, aerobic, and motile. The G + C content of DNA is 61.4 mol%. The fatty acid composition and phylogenetic analysis of the 16S rRNA gene sequence of strain S23 showed that it is related to the members of the genus Comamonas, and most closely related to Comamonas testosteroni (99.9% sequence similarity). The isolate S23 exhibited thiosulfate oxidation under a mixotrophic growth condition. Polymerase chain reaction (PCR) using soxB-specific primers and DNA sequencing showed the presence of the soxB gene. This is the first report in Comamonas sp. showing thiosulfate oxidation under a mixotrophic growth condition. PMID- 19189182 TI - Passive protection assay of monoclonal antibodies against dengue virus in suckling mice. AB - Dengue fever and dengue hemorrhagic fever/dengue shock syndrome are highly infectious diseases caused by dengue virus (DV). Specific monoclonal antibodies (mAbs) against DV are vital for diagnosis, pathological studies, and passive immune therapy. In this study, purified DV serotype 2 (DV2) was used as antigen and BALB/c mice were immunized to induce specific antibodies. We established five hybridoma cell lines, called 78#, 1E7, 7F7, 8F12, and 8H1, respectively, and evaluated them by enzyme-linked immunosorbent assay, indirect immunofluorescence assay, Western blot, plaque reduction neutralization test, and suckling mice protection assay. Lines 78#, 1E7, 7F7, and 8F12 showed a neutralizing effect, and lines 78#, 1E7, 8F12, and 8H1 recognized envelope glycoprotein of DV2. Among them, lines 78# and 8F12 had stronger neutralizing ability in vitro and could protect some suckling mice from virus challenge. Our results demonstrate that immunization with purified virion is efficient for the production of specific neutralizing mAbs against DV2, and these mAbs could be useful tools for studying or treating DV infection. PMID- 19189183 TI - Marinobacterium marisflavi sp. nov., isolated from a costal seawater. AB - A marine bacterium designated strain IMCC4074(T) was isolated from surface seawater collected off Incheon Port, the Yellow Sea, and subjected to a polyphasic taxonomy. The strain was Gram-negative, chemoheterotrophic, slightly halophilic, strictly aerobic, and motile rods. Based on 16S rRNA gene sequence comparisons, the strain was most closely related to Marinobacterium litorale KCTC 12756(T) (93.9%) and shared low 16S rRNA gene sequence similarities with members of the genus Marinobacterium (91.8-93.9%) and the genus Neptunomonas (93.4%) in the order Oceanospirillales. Phylogenetic analyses showed that this marine isolate formed an independent phyletic line within the genus Marinobacterium clade. The DNA G+C composition of the strain was 56.0 mol% and the predominant constituents of the cellular fatty acids were C(16:0) (28.0%), C(16:1 )omega7c and/or iso-C(15:0) 2-OH (19.3%), C(18:1 )omega7c (17.8%), and C(17:1) cyclo (12.5%), which differentiated the strain from other Marinobacterium species. Based on the taxonomic data collected in this study, only a distant relationship could be found between strain IMCC4074(T) and other members of the genus Marinobacterium, thus the strain represents a novel species of the genus Marinobacterium, for which the name Marinobacterium marisflavi sp. nov. is proposed. The type strain of Marinobacterium marisflavi is IMCC4074(T) (= KCTC 12757(T) = LMG 23873(T)). PMID- 19189184 TI - Unconventional integration of the bla gene from plasmid pIT2 during ISlacZ/hah transposon mutagenesis in Pseudomonas aeruginosa PAO1. AB - The ISlacZ/hah transposon carried by pIT2 and derived originally from Tn5 has been a popular system in the generation of random insertion mutants of Pseudomonas aeruginosa. Using this system in the current study, two transconjugants were identified as conferring high levels of carbenicillin resistance. Analyses by gene complementation tests and site-specific gene knockout experiments support the conclusion that carbenicillin resistance in these two mutants is not due to the insertion of ISlacZ/hah transposon into the affected genes. Instead, the production of a TEM beta-lactamase was detected, and integration of the bla gene from pIT2 to the chromosome of the recipient strain was confirmed by polymerase chain reaction. This surprising event was reproducible, with an estimated frequency among the transconjugants of 4% to 10%, and it may cause a potential complication in the interpretation of mutant phenotypes without notice. PMID- 19189185 TI - Sex pheromone components of the pear fruit moth, Acrobasis pyrivorella (Matsumura). AB - We analyzed the sex pheromone of the pear fruit moth, Acrobasis pyrivorella, by means of gas chromatography-electroantennographic detection (GC-EAD) and GC-mass spectrometry. Two EAD-active compounds were detected in the pheromone gland extract of females. They were identified as (Z)-9-pentadecenyl acetate (Z9 15:OAc) and pentadecyl acetate (15:OAc). The amounts per female gland (mean +/- standard error) of these compounds were 12.9 +/- 2.8 and 0.8 +/- 0.1 ng, respectively. Synthetic Z9-15:OAc (300 microg) attracted conspecific males in field trapping experiments. When 15:OAc (21 microg; 7% of Z9-15:OAc quantity) was added, the number of males trapped increased significantly. Catch in traps baited with the mixture of these compounds was greater than that in traps baited with 1 3-day-old virgin females. We, therefore, conclude that Z9-15:OAc and 15:OAc are sex pheromone components of this species. PMID- 19189186 TI - Surgical factors influence the outcome after Ivor-Lewis esophagectomy with intrathoracic anastomosis for adenocarcinoma of the esophagogastric junction: a consecutive series of 240 patients at an experienced center. AB - BACKGROUND: Despite a considerable number of randomized studies, the surgical approach to locally advanced adenocarcinoma of the esophagogastric junction (AEG) I and II is still discussed controversially. Thus, we evaluated the surgical risk and outcome after an abdominothoracic esophagectomy (Ivor-Lewis) with intrathoracic anastomosis as standard procedure. METHODS: Between 1998 and 2006, a total of 240 consecutive patients underwent standardized right thoracoabdominal esophagectomy with two-field lymphadenectomy and intrathoracic anastomosis (Ivor Lewis operation) for AEG I (n = 206) or AEG II (n = 34). A total of 157 patients (65.4%) had neoadjuvant chemotherapy. RESULTS: Postoperative morbidity occurred in 17.9% (43 of 240). Overall mortality was 3.8% (9 of 240). The majority of patients (4 of 9) died because of severe pulmonary complications (44.4%) irrespective of surgical complications. Neoadjuvant chemotherapy did not increase morbidity or mortality. The median overall survival was 51 months. Multivariate analysis including age >75 years, clinical response to chemotherapy, complications, R-category and N-category revealed R-category (P = .005; relative risk [RR] 0.32, 95% confidence interval [95% CI] 0.14-0.70) and complications (P < .001, RR 0.16, 95% CI 0.08-0.35) as independent prognostic factors for all patients. Complications was the only independent prognostic factor (P < .001, RR 0.09, 95% CI 0.08-0.35) for the R0 resected patients. CONCLUSIONS: At an experienced center, Ivor-Lewis resection is a safe surgical procedure. Outcome of patients was significantly influenced by surgical factors such as complete resection and complications. Neoadjuvant chemotherapy did not lead to higher morbidity and mortality. The high mortality from non-surgery-related complications emphasizes the importance of careful preoperative evaluation of comorbidities and patient selection. PMID- 19189187 TI - Long-term oncologic results and complications after preoperative chemoradiotherapy for rectal cancer: a single-institution experience after a median follow-up of 95 months. AB - BACKGROUND: This study sought to evaluate the long-term outcome and complications, and occurrence of second malignancy after preoperative chemoradiotherapy (pCRT) for rectal cancer. METHODS: One hundred twenty-three consecutive patients (78 men, 45 women) with locally advanced mid-low rectal cancer underwent pCRT between 1994 and 2002. Patients were followed up by one surgeon with a standard protocol, and data were prospectively recorded in a dedicated database. No patient was lost to follow-up. Complications were defined as late if they occurred >6 months after surgery. Overall and disease-free survival were calculated by the Kaplan-Meier method. RESULTS: Of 123 patients, 111 underwent an R0 procedure. The rate of pathologic complete response was 16% (n = 20 patients). At a median follow-up of 95 (range, 56-160) months, 50 late complications occurred in 41 patients, 21 of whom required surgery. In seven cases, the complications were clearly CRT related and were significantly associated with the total dose of radiation delivered (P < .05). The estimated 5- and 10-year overall survival was 76% and 67%, respectively. The estimated 5- and 10-year disease-free survival was 83% and 82%, respectively. In 18 of 19 patients who experienced recurrence (local, n = 3; distant, n = 16), it occurred within 48 months from surgery. The most frequent site of metastasis as first site of recurrence was the lung (9 of 19). The most frequent second primary malignancy was lung cancer (3 of 8). CONCLUSIONS: Despite satisfactory oncological outcome, late morbidity after pCRT is relevant and related to the radiotherapy dose used. Most recurrences and second malignancies were located in the lung. PMID- 19189188 TI - Extent of thyroidectomy and lymphadenectomy in 254 patients with papillary thyroid microcarcinoma: a single-institution experience. AB - BACKGROUND: No agreement has been made about the optimal extent of thyroidectomy or lymph node dissection in papillary thyroid microcarcinoma (PTMC). Our aim was to find out the factors associated with the presence of lymph node metastases in the patients with PTMC and to discuss the extent of thyroidectomy and lymphadenectomy. METHODS: A total of 254 patients with PTMC (212 women, 42 men; age 14 to 85 years, median 47 years) were treated at our institute between 1975 and 2007. Lymph node metastases were confirmed in 59 patients (23%) by histopathology. The median observation time was 56 (range, 1-397) months, and the recurrence was diagnosed in seven patients. Modified radical neck dissection and central neck dissection were performed in 55 and 30 patients, respectively. The data on the clinicopathological characteristics of patients and treatment were collected. RESULTS: An incidental PTMC was diagnosed in 107 patients postoperatively. In none of them was a lymph node metastasis or tumor recurrence detected. Preoperatively, an overt PTMC was diagnosed in 147 patients. Clinically manifest metastases were diagnosed in 59 (40%) of them: in 51 preoperatively, and in 8 during the surgical procedure. In 147 patients who had preoperative diagnosis of PTMC, the factors that correlated (P < 0.05) with the presence of lymph node metastases were sex, age, and tumor type. CONCLUSIONS: The patients with high-risk PTMC with preoperatively confirmed lymph node metastases had a higher recurrence rate than those with low-risk PTMC without preoperatively detectable lymph node metastases; therefore, careful therapeutic node dissection is necessary in high-risk patients. PMID- 19189189 TI - Optimizing regional therapy for melanoma. PMID- 19189190 TI - Follow-up of melanoma patients: the need for evidence-based protocols. PMID- 19189191 TI - Prognostic value of CEA and CA 19-9 tumor markers combined with cytology from peritoneal fluid in colorectal cancer. AB - BACKGROUND: Early diagnosis and management of peritoneal metastases from colorectal cancer patients are difficult clinical challenges. The aims of this study were to evaluate the clinical significance of tumor markers and cytology in peritoneal effusions (PE) and peritoneal irrigation fluid (PI) and to determine their value as prognostic indicators in this disease. METHODS: Two hundred thirty four consecutive patients who underwent abdominal surgery for colorectal cancer from January 2006 to December 2007 were included, and tumor markers and cytology in PE and PI were analyzed prospectively. RESULTS: The incidence of free cancer cells retrieved from peritoneal samples was 7.9%. Cytology was positive in 40.0% by Papanicolaou and Giemsa staining, 73.3% by hematoxylin and eosin staining of cell blocks, and 66.7% by carcinoembryonic antigen (CEA) and calretinin immunohistochemistry. Multivariate analysis revealed that peritoneal CEA and cancer antigen (CA) 19-9 in PI were correlated with peritoneal metastasis and cytology. Level of peritoneal fluid CEA was statistically significantly correlated with recurrence and peritoneal metastatic recurrence in patients with negative peritoneal cytology. Cytology, peritoneal CEA, and peritoneal CA 19-9 showed correlations with cancer-free survival and overall survival. CONCLUSIONS: These correlations demonstrate the importance of continuous follow-up of peritoneal metastasis if there is positive cytology or an increase in CEA and CA 19-9 in peritoneal fluid. PMID- 19189192 TI - National survey of patients' bill of rights statutes. AB - BACKGROUND: Despite vigorous national debate between 1999-2001 the federal patients' bill of rights (PBOR) was not enacted. However, states have enacted legislation and the Joint Commission defined an accreditation standard to present patients with their rights. Because such initiatives can be undermined by overly complex language, we surveyed the readability of hospital PBOR documents as well as texts mandated by state law. METHODS: State Web sites and codes were searched to identify PBOR statutes for general patient populations. The rights addressed were compared with the 12 themes presented in the American Hospital Association's (AHA) PBOR text of 2002. In addition, we obtained PBOR texts from a sample of hospitals in each state. Readability was evaluated using Prose, a software program which reports an average of eight readability formulas. RESULTS: Of 23 states with a PBOR statute for the general public, all establish a grievance policy, four protect a private right of action, and one stipulates fines for violations. These laws address an average of 7.4 of the 12 AHA themes. Nine states' statutes specify PBOR text for distribution to patients. These documents have an average readability of 15th grade (range, 11.6, New York, to 17.0, Minnesota). PBOR documents from 240 US hospitals have an average readability of 14th grade (range, 8.2 to 17.0). CONCLUSIONS: While the average U.S. adult reads at an 8th grade reading level, an advanced college reading level is routinely required to read PBOR documents. Patients are not likely to learn about their rights from documents they cannot read. PMID- 19189193 TI - The contribution of cancer incidence, stage at diagnosis and survival to racial differences in years of life expectancy. AB - BACKGROUND: African Americans have higher cancer mortality rates than whites. Understanding the relative contribution of cancer incidence, stage at diagnosis and survival after diagnosis to the racial gap in life expectancy has important implications for directing future health disparity interventions toward cancer prevention, screening and treatment. OBJECTIVE: We estimated the degree to which higher cancer mortality among African Americans is due to higher incidence rates, later stage at diagnosis or worse survival after diagnosis. DESIGN: Stochastic model of cancer incidence and survival after diagnosis. PATIENTS: Surveillance and Epidemiology End Result cancer registry and National Health Interview Survey data. MEASUREMENTS: Life expectancy if African Americans had the same cancer incidence, stage and survival after diagnosis as white adults. RESULTS: African American men and women live 1.47 and 0.91 fewer years, respectively, than whites as the result of all cancers combined. Among men, racial differences in cancer incidence, stage at diagnosis and survival after diagnosis account for 1.12 (95% CI: 0.52 to 1.36), 0.17 (95% CI: -0.03 to 0.33) and 0.21 (95% CI: 0.05 to 0.34) years of the racial gap in life expectancy, respectively. Among women, incidence, stage and survival after diagnosis account for 0.41 (95% CI: -0.29 to 0.60), 0.26 (95% CI: -0.06 to 0.40) and 0.31 (95% CI: 0.05 to 0.40) years, respectively. Differences in stage had a smaller impact on the life expectancy gap compared with the impact of incidence. Differences in cancer survival after diagnosis had a significant impact for only two cancers-breast (0.14 years; 95% CI: 0.05 to 0.16) and prostate (0.05 years; 95% CI 0.01 to 0.09). CONCLUSIONS: In addition to breast and colorectal cancer screening, national efforts to reduce disparities in life expectancy should also target cancer prevention, perhaps through smoking cessation, and differences in survival after diagnosis among persons with breast and prostate cancer. PMID- 19189194 TI - Providers' experiences treating chronic pain among opioid-dependent drug users. AB - BACKGROUND: Successful management of chronic pain with opioid medications requires balancing opioid dependence and addiction with pain relief and restoration of function. Evaluating these risks and benefits is difficult among patients with chronic pain and pre-existing addiction, and the ambiguity is increased for patients on methadone maintenance therapy for opioid dependence. Providers treating both chronic pain and addiction routinely make diagnostic and therapeutic decisions, but decision-making strategies in this context have not been well described. OBJECTIVE: Our objective was twofold. We sought first to explore providers' perceptions of ambiguity, and then to examine their strategies for making diagnostic and treatment decisions to manage chronic pain among patients on methadone maintenance therapy. DESIGN: Qualitative semi-structured interviews. SETTING AND PARTICIPANTS: We interviewed health-care providers delivering integrated medical care and substance abuse treatment to patients in a methadone maintenance program. RESULTS: Providers treating pain and co-morbid addiction described ambiguity in all diagnostic and therapeutic decisions. To cope with this inherent ambiguity, most providers adopted one of two decision making frameworks, which determined clinical behavior. One framework prioritized addiction treatment by emphasizing the destructive consequences of abusing illicit drugs or prescription medications; the other prioritized pain management by focusing on the destructive consequences of untreated pain. Identification with a decision-making framework shaped providers' experiences, including their treatment goals, perceptions of treatment risks, pain management strategies, and tolerance of ambiguity. Adherence to one of these two frameworks led to wide variation in pain management practices, which created tension among providers. CONCLUSIONS: Providers delivering integrated medical care and substance abuse treatment to patients in a methadone maintenance program found tremendous ambiguity in the management of chronic pain. Most providers adopted one of the two divergent heuristic frameworks we identified, which resulted in significant variations in pain management. To reduce variation and determine best practices, studies should examine clinically relevant endpoints, including pain, illicit drug use, prescription drug abuse, and functional status. Until then, providers managing chronic pain in patients with co-morbid addiction should attempt to reduce tension by acknowledging ambiguity and engaging in open discourse. PMID- 19189195 TI - Editorial: The Association of Bone and Joint Surgeons, Orthopaedia, and you. PMID- 19189196 TI - Serological survey of avian H5N2-subtype influenza virus infections in human populations. AB - To investigate the distribution of antibodies against H5N2 influenza virus in humans living in Ibaraki prefecture, Japan, 266 single serum samples were collected to perform serological tests. Results were compared to investigate the relationship between positive results and several factors. The number of positive serum neutralization antibody titers (> or = 40) against avian influenza virus A/H5N2 was significantly greater (P < 0.05) among poultry workers, in comparison to a Japanese healthy population. The geometric mean titers of serum neutralization antibody against A/H5N2 were significantly higher (P < 0.05) among Ibaraki inhabitants and poultry workers (P < 0.0001) when compared to a Japanese healthy population. Seropositivity against A/H5N2 virus was significantly (P < 0.05) associated with age (> or = 50 years old) in poultry workers. These results suggest that seropositivity against H5N2 virus in Ibaraki specimens is significantly higher than those of a Japanese healthy population and that the surveillance of avian influenza viruses is very important to evaluate the invasion or emergence of new pandemic influenza viruses from species other than humans. PMID- 19189197 TI - Multiorgan distribution of human influenza A virus strains observed in a mouse model. AB - Multiorgan spread and pathogenesis of influenza infection with three human influenza A viruses was studied in mice. Mouse-adapted viruses A/Dunedin/4/73(H3N2), A/Mississippi/1/85(H3N2), and A/PR/8/34(H1N1) differed considerably in virulence (p.f.u./LD(50)): 79,000 p.f.u. for Dunedin, 5,000 p.f.u. for Mississippi, and 65 p.f.u. for PR/8, which qualified Dunedin as low virulent, Mississippi as intermediate, and PR/8 as highly virulent. All three viruses were detected in lungs, heart, and thymus by cultivation and RT-PCR. Moreover, vRNA of all viruses was found in liver and spleen, of Dunedin and PR/8 also in kidneys and that of Dunedin and Mississippi in blood. Only vRNA of Dunedin was demonstrated in brain. Lung damage accompanied by histopathological changes and thymus reduction were most extensive after infection with the highly virulent virus PR/8. We assume that the ability to spread to multiple organs may be a more common property of influenza viruses in mammalian hosts than previously believed. PMID- 19189198 TI - Complete genomic sequences of Tomato yellow leaf curl Mali virus isolates infecting tomato and pepper from the North Province of Cameroon. AB - Complete genomic sequences of Tomato yellow leaf curl Mali virus isolates infecting tomato and pepper from the North Province of Cameroon. PMID- 19189199 TI - L-Glutamine or L-alanyl-L-glutamine prevents oxidant- or endotoxin-induced death of neonatal enterocytes. AB - This study tested the hypothesis that L-glutamine (Gln) or L-alanyl-L-glutamine (Ala-Gln) prevents oxidant- or endotoxin-induced death of neonatal enterocytes. Enterocytes of neonatal pigs rapidly hydrolyzed Ala-Gln and utilized Gln. To determine whether Gln or Ala-Gln has a cytoprotective effect, IPEC-1 cells were cultured for 24 h in Gln-free Dulbecco's modified Eagle's-F12 Ham medium containing 0, 0.5, 2.0 or 5.0 mM Gln or Ala-Gln, and 0, 0.5 mM H(2)O(2) or 30 ng/ml lipopolysaccharide (LPS). Without Gln or Ala-Gln, H(2)O(2)- or LPS-treated cells exhibited almost complete death. Gln or Ala-Gln at 0.5, 2 and 5 mM dose dependently reduced H(2)O(2)- or LPS-induced cell death by 14, 54 and 95%, respectively, whereas D: -glutamine, alanine, glutamate, ornithine, proline, glucosamine or nucleosides had no effect. To evaluate the effectiveness of Gln or Ala-Gln in vivo, 7-day-old piglets received one-week oral administration of Gln or Ala-Gln (3.42 mmol/kg body weight) twice daily and then a single intraperitoneal injection of LPS (0.1 mg/kg body weight); piglets were euthanized in 24 and 48 h to analyze intestinal apoptotic proteins and morphology. Administration of Gln or Ala-Gln to LPS-challenged piglets increased Gln concentrations in small-intestinal lumen and plasma, reduced intestinal expression of Toll-like receptor-4, active caspase-3 and NFkB, ameliorated intestinal injury, decreased rectal temperature, and enhanced growth performance. These results demonstrate a protective effect of Gln or Ala-Gln against H(2)O(2)- or LPS-induced enterocyte death. The findings support addition of Gln or Ala-Gln to current Gln-free pediatric amino acid solutions to prevent intestinal oxidative injury and inflammatory disease in neonates. PMID- 19189200 TI - Tyrosine-kinases in bacteria: from a matter of controversy to the status of key regulatory enzymes. AB - When considering protein phosphorylation in bacteria, phosphorylation of aspartic acid and histidine residues mediated by the two-component systems is the first to spring to mind. And yet other phosphorylation systems have been described in bacteria in the past 20 years including eukaryotic-like serine/threonine kinases and more recently tyrosine-kinases. Among the latter, a peculiar type is widespread among bacteria, but not in higher organisms. These enzymes possess unique structural features defining thus a new family of enzymes termed Bacterial tyrosine kinases (BY-kinases). BY-kinases have been shown to be mainly involved in polysaccharide production, but their ability to phosphorylate endogenous substrates indicates that they participate in the regulation of other functions of the bacterial cell. Recent advances in mass spectrometry based phosphoproteomics provided lists of many new phosphotyrosine-proteins, indicating that BY-kinases may be involved in regulating a large array of other cellular functions. One may expect that in a near future, tyrosine phosphorylation will turn out to be one of the key regulatory processes in the bacterial cell and will yield new insights into the understanding of its physiology. PMID- 19189202 TI - Mitotic drivers--inhibitors of the Aurora B Kinase. AB - In this article we review the basis for current anti-mitotic, anti-cancer, therapy and the potential for Aurora B kinase inhibitors as a new differentiated class of agents--"mitotic drivers". We review the current understanding of Aurora B inhibition from basic cell biology to inhibitors currently undergoing clinical trials. PMID- 19189201 TI - Cloning and expression of the tumstatin active peptides-T(7) and its derivant T(7)-NGR. AB - To enhance the role targeting, design to link NGR sequence with tumstatin active peptides-T(7)'s C-terminal, the derivant called T(7)-NGR. The cloning vector pMD T(7) and pMD-T(7) N were constructed by PCR and gene synthesis methods, respectively, identified by digestion and DNA sequencing. After the digested plasmids were isolated by the low melting point agarose electrophoresis, the target-fragment was cut off and mixed with the recovery of the digested vector pET28a. Expression vector pET-T(7) and pET-T(7) N were constructed in low melting point agarose, identified by digestion and DNA sequencing, transformed into competent Escherichia coli BL21 (DE3), induced by IPTG. Identification result shows that pET-T(7) and pET-T(7) N were correct. Tricine-SDS-PAGE results showed that IPTG concentration of 1 mM, after the induction of 25 degrees C, 8 h, T(7) peptides and T(7)-NGR peptides have achieved the optimum conditions of expression. In conclusion, the expression vectors of the two peptides has been successfully constructed, and got product, no coverage at home and abroad, laid the foundation for further activity experiments. PMID- 19189203 TI - Properties influencing the relative binding affinity of pteroate derivatives and drug conjugates thereof to the folate receptor. AB - PURPOSE: Using in vitro competition assays, determine salient chemical features of pteroates and pteroate-drug conjugates which afford high affinity to the folate receptor. MATERIALS AND METHODS: Both folate binding protein-coated polystyrene plates and adherent human cell-based assays were used to evaluate the effects of assay temperature and buffer composition on pteroate/pteroate-drug conjugate binding affinity. Following assay selection and optimization, the relative binding affinities of ten vitamers and derivatives as well as seven pteroate-drug conjugates were evaluated. RESULTS: Compared to polystyrene plates containing immobilized folate binding protein, adherent KB cells were determined to be an equally effective, more desirable source of folate receptor for such analyses. Using the latter method, we discovered that a charged group positioned in close proximity to the pteroate's aryl moiety is critical for retaining high binding affinity. We also found that a diverse set of bioactive small molecule agents can be attached to folic acid in a manner that does not appreciably disturb this vitamin's intrinsic high affinity for the folate receptor. However, conjugation of lipophilic, high protein-binding agents to folate was sometimes found to dramatically reduce affinity, which is a finding that best exemplifies the need for having a reliable in vitro assay for determining a compound's RA. CONCLUSION: Molecules which bind best to the human folate receptor are those that contain hydrophilic regions distal to the ligand's aryl group, and for drug conjugates, an extended hydrophilic spacer placed in-between the pteroate and drug cargo moieties. PMID- 19189204 TI - An investigation into the rheology of pharmaceutical inter-granular material bridges at high shear rates. AB - PURPOSE: This study was undertaken to investigate the rheological properties of inter-granular material bridges on the nano-scale when strained at high shear rates. MATERIALS AND METHODS: Atomic force microscopy (AFM) was used as a rheometer to measure the viscoelasticity of inter-granular material bridges for lactose:PVP K29/32 and lactose:PVP K90 granules, produced by wet granulation. RESULTS: The loss tangent (tan delta) and both the storage (G') and loss shear moduli (G'') of inter-granular material bridges were measured as a function of the probe-sample separation distance, oscillation frequency and relative humidity (RH). As the probe was withdrawn from the granule surface tan delta initially increased rapidly from zero to a plateau phase. G'' became increasingly dominant as the bridge was further extended and eventually exceeded G'. At high RH, capillary forces were foremost at bridge rupture, whereas at low RH elastic forces dominated. The effect of increasing frequency was to increase the effective elasticity of the bridge at high RH. CONCLUSIONS: AFM has been employed as a rheometer to investigate the nano-scale rheology of inter-granular material bridges. This novel method may be used to obtain a fundamental understanding how different binders, granulated with different diluent fillers, behave at high shear rates. PMID- 19189205 TI - CD8+ cell anti-HIV activity rapidly increases upon discontinuation of early antiretroviral therapy. AB - INTRODUCTION: CD8+ lymphocytes can suppress HIV replication without killing the infected cells. This CD8+ cell noncytotoxic anti-HIV response (CNAR) is associated with a beneficial clinical course. MATERIALS AND METHODS: In this longitudinal study of 16 participants in the Options Project at UCSF, we measured the ability of CD8+ lymphocytes to suppress HIV replication in CD4+ cells during primary HIV infection, early antiretroviral therapy, and after treatment. RESULTS AND DISCUSSION: CD8+ lymphocytes from subjects with untreated primary HIV-1 infection strongly suppressed HIV replication. Initiation of antiretroviral therapy during primary HIV-1 infection caused a marked decline in this CNAR. CD8+ cells from these subjects regained anti-HIV activity when early therapy was discontinued. The timing of the appearance of CD8+ cell anti-HIV activity directly correlated with the emergence of detectable virus levels. Maximal CNAR activity coincided with a decay in the kinetics of HIV replication. In addition, peak viral loads during treatment interruption were lower than pre-treatment virus levels (median reduction = 0.8 logs, p = 0.005) and CD4+ T cell counts were maintained for a 24-week period of follow-up. CONCLUSION: These results suggest that CNAR plays an important role in suppressing HIV replication in the setting of antiretroviral treatment interruption in HIV-infected individuals. PMID- 19189206 TI - Interaction of curcumin and diacetylcurcumin with the lipocalin member beta lactoglobulin. AB - The binding of curcumin (CUR) and diacetylcurcumin (DAC) to bovine beta lactoglobulin (BLG) genetic variant B was investigated by fluorescence and circular dichroism techniques. The binding parameters including number of substantive binding sites and the binding constants have been evaluated by fluorescence quenching method. The distance (r) between donor (BLG) and acceptor (CUR and DAC) was obtained according to the Forster's theory of non-radiative energy transfer. The far-UV circular dichroism spectra were used to investigate the possible changes in the secondary structure of BLG in the presence of CUR and DAC and showed that these two ligands change the alpha-helix and random coil contents of this protein to some extent. The visible circular dichroism spectra indicated that the optical activity during the ligand binding was observed due to the induced-protein chirality. All of the achieved results suggested the important role of the phenolic OH group of CUR in the binding process resulted in more affinity of CUR than DAC for binding to BLG. PMID- 19189207 TI - Erythrocyte total carbonic anhydrase esterase activity in african american obese children: reduction starts at a young age. PMID- 19189208 TI - Genetic variation and association of insulin-like growth factor binding protein-3 with performance in Swine. AB - Genetic variation of insulin-like growth factor binding protein-3 was analyzed in 17 pig breeds (14 native Chinese and 3 European). Using PCR-single strand conformation polymorphism, we found a polymorphism in intron 2, and this SNP was the combined mutation of G897T-G903A-C911T. The Chinese breeds carried a higher TAT/TAT genotype frequency (over 50%), except for Bamei (22%), Yujiang Black (0.0%), and Erhualian (10.0%); the European breeds had a higher GGC/GGC genotype frequency (Large White 1.67%, Landrace 13.89%, Duroc 0.0%). The allelic frequency of TAT in Chinese breeds was over 50%, except for Yujiang Black (12.5%); the allelic frequency of GGC was over 50% in all European breeds. The effect of genotype on 43 performance traits was investigated in one population (Lantang x Landrace). Pigs with the TAT/TAT genotype had higher B-point and C-point back-fat thickness than pigs with the GGC/GGC genotype. The TAT/TAT pigs also scored higher in meat color than the GGC/GGC pigs. These results implied that IGFBP-3 may affect meat quality and carcass traits. PMID- 19189209 TI - Novel polymorphism at the 3' UTR of the caprine adiponectin gene. PMID- 19189210 TI - Impact of CYP2D6 polymorphisms in tamoxifen adjuvant breast cancer treatment. AB - The aim of this study is to evaluate the impact of CYP2D6 genotyping in predicting disease-free survival and toxicity in breast cancer patients treated with adjuvant tamoxifen. DNA from 91 patients was genotyped using the AmpliChip CYP450 GeneChip, Roche that facilitates the classification of individuals by testing 27 alleles. When patients were grouped into group 1 (*4/*4, *4/*41, *1/*5 and *2/*5) and group 2 (the remaining genotypes), a significant difference in disease-free survival (DFS) was observed between groups (P = 0.016). The mean DFS in group 1 was 95 months in contrast with 119 months in group 2. No significant relationship was found between the CYP2D6 genotype classification and severe, mild or no toxicity (P = 0.2). Nevertheless, severe, and mild toxicity was more frequent among poor metabolizer patients than in patients with a normal metabolizer pattern (18.8 and 43.8% vs. 10.7 and 36%, respectively). In breast cancer, patients treated with adjuvant tamoxifen, non-functional and severely impaired CYP2D6 variants are associated with a worse DFS and with a higher frequency of severe and mild toxicities. Larger studies of the CYP2D6 genotype clinical outcomes association are needed to complement initial results. PMID- 19189211 TI - Patterns of recurrence in the basal and non-basal subtypes of triple-negative breast cancers. AB - Traditional prognostic markers for breast cancer include estrogen receptor (ER), progesterone receptor (ER) and HER2/neu. Negative staining for these three markers defines the 'triple-negative' phenotype. By adding markers for cytokeratin 5/6 and EGFR, triple-negative breast cancers can be divided into 'basal-like' and 'normal-like' subgroups. We conducted immuno-staining on a panel of 958 patients with breast cancer, using all five markers and we followed the patients for distal recurrence and death. We compared rates of distal recurrence in the basal-like and normal-like subgroups with that of women with ER-positive breast cancer. Only 16 of 958 women had normal-like breast cancers. These cancers resembled basal-like cancers in that they had a high proliferative index, but the women with normal-like breast cancers resembled ER-positive women in terms of distant recurrence. The addition of CK5/6 and EGFR to the standard panel (ER/PR/HER2/neu) defines a small subgroup of women with normal-like breast cancer. The prognosis of these women may be superior to that of basal-like breast cancers but firm conclusions cannot be made. PMID- 19189212 TI - The CYP2D6*4 polymorphism affects breast cancer survival in tamoxifen users. AB - Cytochrome P450 2D6 (CYP2D6) plays an important role in the formation of endoxifen, the active metabolite of tamoxifen. In this study the association between the most prevalent CYP2D6 null-allele in Caucasians (CYP2D6*4) and breast cancer mortality was examined among all incident users of tamoxifen in a population-based cohort study. Breast cancer mortality was significantly increased in patients with the * 4/*4 genotype (HR = 4.1, CI 95% 1.1-15.9, P = 0.041) compared to wild type patients. The breast cancer mortality increased with a hazard ratio of 2.0 (CI 95% 1.1-3.4, P = 0.015) with each additional variant allele. No increased risk of all-cause mortality or all-cancer mortality was found in tamoxifen users carrying a CYP2D6*4 allele. The risk of breast cancer mortality is increased in tamoxifen users with decreased CYP2D6 activity, consistent with the model in which endoxifen formation is dependent on CYP2D6 activity. PMID- 19189213 TI - Frequent somatic mutations of GATA3 in non-BRCA1/BRCA2 familial breast tumors, but not in BRCA1-, BRCA2- or sporadic breast tumors. AB - Heterozygous somatic mutations of the transcription factor, GATA-3, have recently been reported in approximately 5% breast of tumors unselected for family history. We sequenced the GATA-3 gene in 55 breast tumors from women with familial breast cancer, and found seven heterozygous somatic mutations, all in non-BRCA1/2 cases in which the frequency was 22%. In contrast, we found mutations of GATA-3 in only 4% of 81 sporadic tumors analysed. It is possible that GATA3 mutations occur earlier in the evolution of BRCAx tumors, compared to BRCA1, BRCA2 or sporadic tumors, and are therefore easier to detect by direct sequencing in the presence of some stromal contamination. PMID- 19189214 TI - Spectroscopic and differential scanning calorimetric studies on the unfolding of Trichosanthes dioica seed lectin. Similar modes of thermal and chemical denaturation. AB - Physico-chemical and unfolding studies have been carried out on Trichosanthes dioica seed lectin (TDSL). The lectin exhibited maximum activity between pH 7.0 and 10.0, which decreased steeply at lower pH. The hemagglutination activity of TDSL was unaffected in the temperature range 4-50 degrees C, but decreased rapidly at higher temperatures. Differential scanning calorimetric studies indicate that thermal unfolding of TDSL is an irreversible process, which could be described by a three-state model. The calorimetric scan recorded at pH 7.0 consists of two transitions, occurring at around 338.6 K, and 342.8 K. In the presence of carbohydrate ligands both these transitions shifted to higher temperatures, suggesting that ligand binding stabilizes the native conformation of the protein. The unfolding temperature was highest at pH 5.0 indicating that TDSL is more stable at acidic pH. Gdn.HCl induced unfolding, monitored by following changes in the intrinsic fluorescence properties of the protein, was also observed to be a three-state process involving an intermediate. CD spectroscopy indicates that the secondary and tertiary structures of TDSL are rather similar at different pH values, indicating that the lectin structure remains essentially unchanged over a wide range of pH. PMID- 19189215 TI - Evaluation of pastures for horses grazing on soils polluted by trace elements. AB - Pasture established on polluted soil may pose a risk to grazing livestock creating a requirement for mechanical management which may affect biodiversity and expend energy. The risk associated with managing pasture by grazing horses (non-edible livestock) is being assessed in the Guadiamar Valley (SW Spain), where soils are polluted with trace elements following a major pollution incident. Soil pollution does not affect biomass production or floristic composition of pasture, although both variables influence trace element accumulation in herbage. Element concentrations in herbage are below maximum tolerable limits for horses. Faecal analysis showed regulated absorption of essential elements, while non-essential elements seemed preferentially excreted. Elemental content of horse hair did not differ in animals from polluted and control pastures. If pastures are managed by grazing, periodic monitoring is recommended in view of the long-term chronic trace element exposure in these systems. PMID- 19189216 TI - Correlates of secondhand tobacco smoke exposure among persons with severe and persistent mental illness (SPMI) accessing community mental health services. AB - To examine the correlates of secondhand tobacco smoke (SHS) exposure in a sample of individuals with severe and persistent mental illness (SPMI) accessing community mental health services. Cross-sectional data were collected from a voluntary sample of 788 individuals with SPMI. Nonsmokers were more likely to be exposed on the street or at a bus stop, in a park or public place; whereas smokers were more likely to report SHS exposure where they lived and at their friend's homes. In a multivariate model, only number of acquaintances who smoked and the number of sources of SHS exposure remained significantly associated with perceived frequency of SHS exposure when accounting for other correlates. There is a need for the enactment and enforcement of policies to curtail SHS exposure in outdoor, public, settings. Future research will be necessary to better understand the extent and impact of SHS exposure in this population. PMID- 19189217 TI - Perceived discrimination among three groups of refugees resettled in the USA: associations with language, time in the USA, and continent of origin. AB - The objectives of this study were to assess the prevalence and predictors of discrimination among a community-based sample of refugees resettled in the USA. We sought to test whether language, gender, time in the USA and country of origin were associated with the experience of discrimination among individuals resettled in the USA as part of the refugee resettlement program. Perceived discrimination was assessed among individuals from East Africa (n = 92), West Africa (n = 74), and from Eastern Europe (n = 112) using a multi-item measure of discrimination. Bivariate associations revealed statistically significant associations between experiences of discrimination and time in the USA, language ability, and sending country. A logistic regression model revealed that refugees from African sending countries were more likely than Eastern European individuals to experience discrimination, even after controlling for potentially confounding factors. We interpret this finding as evidence of racism and discuss the implications for population health and resettlement practice. PMID- 19189218 TI - Effects of lanreotide SR and Autogel on tumor mass in patients with acromegaly: a systematic review. AB - Long-acting somatostatin analogs (SSA) are widely used for the treatment of acromegaly achieving biochemical control of the disease in 50-75% of the patients. One of the goals of the treatment of acromegaly is the control of tumor growth, especially in patients in whom SSAs are used as first-line therapy. Over the recent years, there has been growing evidence that SSAs are able to induce tumor shrinkage in patients with acromegaly. However, most of the data are from patients under treatment with octreotide, either subcutaneously or intramuscularly with long-acting formulation, whereas the data on lanreotide SR or Autogel are very few. Indeed, octreotide and lanreotide, i.e. the two commercially available SSAs, show slight differences in pharmacokinetics and patterns of receptor affinities with potentially different therapeutic effects. We aimed to perform a systematic review of literature data concerning the shrinkage effects of long-acting lanreotide in patients with acromegaly. The analysis was focused on the following issues: differences in shrinkage effects between primary and secondary medical treatment, predictive value of baseline tumor volume and correlation between biochemical control and shrinkage effects. The peer-reviewed medical literature was searched to identify clinical trials studying the effects of lanreotide SR or Autogel on adenoma size in acromegaly. To be included in this analysis, studies had one of the following designs: randomized controlled trial; prospective, nonrandomized trial; retrospective study. Twenty-two studies were found to be eligible for the final analysis, in which tumor size was measured as an end-point for lanreotide treatment. Overall a total of 32.8% of patients experienced a variable degree (from 10 to 77%) of tumor shrinkage during lanreotide SR or Autogel treatment. The analysis showed that tumor shrinkage was more frequent in naive patients as compared with those previously treated by radiotherapy, surgery or drugs other than lanreotide. The data on the correlation between tumor shrinkage and baseline tumor size were discordant, but when baseline tumor size was specified, more than 80% of patients undergoing shrinkage under lanreotide Autogel had macroadenomas. Finally, with lanreotide Autogel there was no evident correlation between biochemical response and tumor shrinkage. Our systematic review of the literature shows that lanreotide particularly when used as first-line therapy is able to quite frequently induce tumor shrinkage in patients with acromegaly. This finding suggests that this drug may have a role in the primary treatment of acromegaly. PMID- 19189219 TI - Inducible expression of functional mu opioid receptors in murine dendritic cells. AB - Opioids are known to exert direct effects on the immune system, and the expression of functional opioid receptors has been reported on several immune cell types. Dendritic cells (DCs) are important inducers and regulators of immune responses. In this study, we investigated whether murine dendritic cells express functional mu opioid receptors (MOR). RT-PCR analysis and double immunofluorescence staining revealed the expression of MOR in activated murine dendritic cells. We also studied the dynamic expression of MOR messenger RNA in murine dendritic cells in response to different Toll-like receptor ligands. Functionally, treatment of DCs with endomorphin 1 (EM1), a specific agonist of MOR, can inhibit the forskolin-induced formation of cyclic adenosine monophosphate level in activated DCs. Moreover, EM1 treatment resulted in less activation of p38 MAPK and more activation of ERK signaling in lipopolysaccharide stimulated DCs. Consistently, treatment of DCs with EM1 altered cytokine production by increasing IL-10 and decreasing IL-12 and IL-23. Our results suggest that MOR is inducibly expressed on activated DCs and functionally mediates EM1-induced effects on DCs. Thus, dendritic cells might be involved in crosstalk between the neuroendocrine and the immune system. PMID- 19189220 TI - Initial characterization of micafungin pulmonary delivery via two different nebulizers and multivariate data analysis of aerosol mass distribution profiles. AB - Pharmaceutical aerosols have been targeted to the lungs for the treatment of asthma and pulmonary infectious diseases successfully. Micafungin (Astellas Pharma US, Deerfield, IL, USA) has been shown to be an effective antifungal agent when administrated intravenously. Pulmonary delivery of micafungin has not previously been reported. In the present pilot study, we characterize the performance of two nebulizers and their potential for delivering micafungin to the lungs as well as the use of multivariate data analysis for mass distribution profile comparison. The concentration of micafungin sodium increased by 21% when delivered by the Acorn II nebulizer and by 20% when delivered by the LC Plus nebulizer, respectively, from the first to the second sampling period. The Acorn II nebulizer delivered a fine particle fraction FPF(5.8) (%<5.8 microm) of 92.5 +/- 0.8 and FPF(3.3) (%<3.3 microm) of 82.3 +/- 2.1 during the first sampling period. For the LC Plus nebulizer, FPF(5.8) was 92.3 +/- 0.1 and FPF(3.3) was 67.0 +/- 0.7 during the first sampling period. The mass median aerodynamic diameter (MMAD) increased from 1.67 +/- 0.05 to 1.77 +/- 0.04 mum (Acorn II nebulizer) and from 2.09 +/- 0.01 to 2.20 +/- 0.01 microm (Pari LC Plus nebulizer) from the first to the second sampling periods. These changes in MMAD were statistically significant by paired t test. Multivariate data analysis showed that this could be explained systematically by greater drug deposition on stages with larger cutoff sizes and reduced drug deposition on stages with smaller cutoff sizes rather than multimodal deposition or other anomalies in size distribution. PMID- 19189222 TI - Technical performance and reflection capacity of the anaesthetic conserving device--a bench study with isoflurane and sevoflurane. AB - OBJECTIVE: The anaesthetic conserving device (AnaConDa), Sedana Medical, Sundbyberg, Sweden) facilitates administration of isoflurane or sevoflurane by liquid infusion. An anaesthetic reflector inside the device conserves exhaled anaesthetic and re-supplies it during inspiration. In this bench study, we examined the influence of infusion rates and ventilatory settings on the resulting anaesthetic concentrations on patient (C(pat)) and ventilator side of the reflector (C(loss)) to describe its technical performance. METHODS: A Puritan Bennett 840 ICU ventilator (Pleasanton, US), AnaConDa, and a test lung (3 l chloroprene-bag) were assembled. Infusion rates (IR, 0.2-50 ml h(-1)), respiratory rates (RR, 5-40 breaths min(-1)), and tidal volumes (V(T), 0.3, 0.5, and 1.0 l) were varied. C(pat) was measured via a thin catheter in the middle of the 3 l-bag in steady state (online data storage and averaging over >10 min). C(loss) was calculated from IR (to yield the volume of vapour per unit of time), and expired minute volume (in which the vapour is diluted) on the assumption that, in the steady state, input by liquid infusion equals output through the reflector. RESULTS: At lower concentrations (C(pat) < 1 vol%) the ratio C(loss)/C(pat) was constant (R(C) = 0.096 +/- 0.012) for all combinations of IR, RR and V(T), both for isoflurane and sevoflurane. The device could efficiently reflect up to 10 ml vapour per breath (e.g. 2 vol% in 0.5 l). When exceeding this capacity, surplus vapour "spilled over" and R(C) markedly increased indicating decreased performance. CONCLUSIONS: The triple product minute volume times R(C) times C(pat) describes anaesthetic losses through the reflector. It can easily be calculated as long as the 10 ml reflection capacity is not exceeded and thus R(C) is constant. Increased minute ventilation necessitates increasing the IR to keep C(pat) constant. When using large V(T) and high C(pat) "spill over" occurs. This effect offers some protection against an inadvertent overdose. PMID- 19189223 TI - When will learning style go out of style? PMID- 19189221 TI - Strengthening the reporting of genetic association studies (STREGA): an extension of the STROBE statement. AB - Making sense of rapidly evolving evidence on genetic associations is crucial to making genuine advances in human genomics and the eventual integration of this information in the practice of medicine and public health. Assessment of the strengths and weaknesses of this evidence, and hence the ability to synthesize it, has been limited by inadequate reporting of results. The STrengthening the REporting of Genetic Association studies (STREGA) initiative builds on the Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) Statement and provides additions to 12 of the 22 items on the STROBE checklist. The additions concern population stratification, genotyping errors, modeling haplotype variation, Hardy-Weinberg equilibrium, replication, selection of participants, rationale for choice of genes and variants, treatment effects in studying quantitative traits, statistical methods, relatedness, reporting of descriptive and outcome data, and the volume of data issues that are important to consider in genetic association studies. The STREGA recommendations do not prescribe or dictate how a genetic association study should be designed but seek to enhance the transparency of its reporting, regardless of choices made during design, conduct, or analysis. PMID- 19189225 TI - Anatomopathological aspects of avian aspergillosis. AB - Aspergillosis is a fungal disease caused by fungi of the genus Aspergillus, in particular A. fumigatus and A. flavus. This paper focuses on anatomopathological aspects resulting from a chronic infection from Aspergillus spp in the chicken (Gallus domesticus), in the herring gull (Larus cachinnans micaelli) and in the red-legged partridge (Alectoris rufa rufa). Microscopically, we observed some histological lesions that are related to the two typical forms of Aspergillosis: a deep nodular form, typical of organs with a non-aerated parenchyma, and a non encapsulated superficial diffuse form typical of the serosae and the lung. The observed forms of aspergillosis have been found in animals raised in poor hygienic environmental conditions or malnourished animals (chicken); in wild birds from wildlife recovery centres (herring gull), which underwent some forms of stress, such as traumas, detention, starvation, extended antibiotic treatments; in game birds (red-legged partridge) used for restocking natural areas that had been negatively affected by such stressors as captivity in aviaries, containment and transport in cages, release in unsuitable environments and malnutrition. The observed anatomopathological and istopathological aspects can therefore be regarded as the outcome of a number of factors that have reduced the typical resistance of the species and impaired the efficiency of their immune systems. PMID- 19189224 TI - Stepwise synthesis of RNA conjugates carrying peptide sequences for RNA interference studies. AB - Oligoribonucleotide conjugates carrying nuclear localization peptide sequences at the 3'-end were prepared stepwise on a single support. The siRNA duplex carrying the nuclear localization peptide sequence at the 3'-end of the passenger strand has similar inhibitory properties as those of unmodified or cholesterol-modified RNA duplexes. PMID- 19189226 TI - Modified annular penile skin flap for repair of pan-anterior urethral stricture. AB - OBJECTIVE: To review our experience with a zigzag shape annular penile fasciocutaneous flap (modified McAninch flap) for reconstruction of pan-anterior urethral strictures. PATIENTS AND METHODS: From 2003 to 2006, 30 male patients with pan-anterior urethral stricture were repaired by single-stage urethroplasty using a modification of the annular penile flap of McAninch for augmentation of a long anterior urethral stricture (zigzag annular penile fasciocutaneous flap). The procedure was total bulbopenile urethral mobilization, stricturotomy, and ventrally positioned zigzag annular penile island fasciocutaneous flap. Age of patients ranged from 20-60 years. Range of stricture length was 15-20 cm. All strictures were bulbopenile post inflammatory or post catheter with exclusion of BXO in all cases by clinical picture and pre-operative biopsy. Follow up period was 24 months. RESULTS: Urethral patency was documented clinically, urodynamically, by RUG, and by urethroscopy for 26 male patients for whom repair was performed by use of this technique. Re-stricture occurred in four patients, fistula in one patient, and post voiding dripping in two patients. CONCLUSIONS: A meticulously fashioned zigzag annular penile skin flap could be raised to bridge a urethral defect up to 20 cm long and has the advantage of one-stage repair. No additional grafts are needed. Cosmetic outcome included a straight penis in all cases. PMID- 19189227 TI - Modification, translation and adaptation of questionnaires: should copyright laws be observed? AB - This commentary is intended to start a discussion about whether people should be allowed to modify, translate, adapt or sell copyrighted questionnaires without the permission of the developer (copyright-holder). PMID- 19189229 TI - A revision of the haploporinae nicoll, 1914 (digenea: haploporidae) from mullets (mugilidae): Saccocoelium Looss, 1902. AB - Saccocoelium Looss, 1902 is revised and a key to its recognised species is presented. S. obesum Looss, 1902 (type-species) and S. tensum Looss, 1902 are redescribed based on material from Liza spp. (Pisces: Mugilidae) in Spanish Mediterranean and, in the case of the former, Bulgarian Black Sea waters. Two new species, S. cephali n. sp. and S. currani n. sp., are described from Mugil cephalus L. in Spanish Mediterranean waters. S. gohari Ramadan, Saoud, Ashour & Mansour, 1989b is recognised and commented upon. Lecithobotrys helmymohamedi Ramadan, Saoud, Ashour & Mansour, 1989a, S. portsaidensis El-Shahawi, El-Gindy, Imam & Al-Bassel, 1992, S. saoudi El-Shahawi, El-Gindy, Imam & Al-Bassel, 1992, Neosaccocoelium aegyptiacus El-Shahawi, El-Gindy, Imam & Al-Bassel, 1992 are considered to be synonyms of S. tensum and Neosaccocoelium El-Shahawi, El-Gindy, Imam & Al-Bassel, 1992 a synonym of Saccocoelium. S. obesum, S. tensum and the two new species are compared morphometrically and distinguished by univariate and multivariate analyses. Lecithobotrys mugilis Rekharani & Madhavi, 1985 is transferred to Unisaccus Martin, 1973 as U. mugilis (Rekharani & Madhavi, 1985) n. comb., and Lecithobotrys sprenti Martin, 1973 [= Saccocoelium sprenti (Martin, 1973) Overstreet & Curran, 2005] is transferred to Unisaccus as U. sprenti (Martin, 1973) n. comb. S. megasacculum Liu, Wang, Peng, Yu & Yang, 2004 is transferred to Elliptobursa Wu, Lu & Zhu, 1996 as E. megasacculum (Liu, Wang, Peng, Yu & Yang, 2004) n. comb. S. tripathi Dutta, 1995 (syn. Saccocoelium tripathi Datta & Manna, 1998) is considered to be a species inquirenda. PMID- 19189228 TI - P2X7 receptors regulate multiple types of membrane trafficking responses and non classical secretion pathways. AB - Activation of the P2X7 receptor (P2X7R) triggers a remarkably diverse array of membrane trafficking responses in leukocytes and epithelial cells. These responses result in altered profiles of cell surface lipid and protein composition that can modulate the direct interactions of P2X7R-expressing cells with other cell types in the circulation, in blood vessels, at epithelial barriers, or within sites of immune and inflammatory activation. Additionally, these responses can result in the release of bioactive proteins, lipids, and large membrane complexes into extracellular compartments for remote communication between P2X7R-expressing cells and other cells that amplify or modulate inflammation, immunity, and responses to tissue damages. This review will discuss P2X7R-mediated effects on membrane composition and trafficking in the plasma membrane (PM) and intracellular organelles, as well as actions of P2X7R in controlling various modes of non-classical secretion. It will review P2X7R regulation of: (1) phosphatidylserine distribution in the PM outer leaflet; (2) shedding of PM surface proteins; (3) release of PM-derived microvesicles or microparticles; (4) PM blebbing; (5) cell-cell fusion resulting in formation of multinucleate cells; (6) phagosome maturation and fusion with lysosomes; (7) permeability of endosomes with internalized pathogen-associated molecular patterns; (8) permeability/integrity of mitochondria; (9) exocytosis of secretory lysosomes; and (10) release of exosomes from multivesicular bodies. PMID- 19189230 TI - A revision of the haploporinae nicoll, 1914 (digenea: haploporidae) from mullets (mugilidae): Dicrogaster Looss, 1902 and Forticulcita overstreet, 1982. AB - The status of the nominal species of Dicrogaster is re-assessed by means of a comparative morphological study based on newly collected material from the western Mediterranean and a critical evaluation of the published data. D. perpusilla Looss, 1902 (type-species) and D. contracta Looss, 1902 are redescribed on the basis of new material from Liza spp. D. fragilis Fernandez Bargiela, 1987 is considered a junior synonym of D. fastigata Thatcher & Sparks, 1958 and D. maryutensis Al-Bassel, 1990 is considered to be nomen nudum. A key to the recognised species of Dicrogaster is presented. A new species of Forticulcita Overstreet, 1982 is described from Mugil cephalus L. in the Spanish Mediterranean and differentiated from the type-species, F. glabra Overstreet, 1982, and the recently described F. mugilis Hassanine, 2007 by its significantly smaller body size and most of its metrical data. The new species, F. gibsoni n. sp., is characterised by a narrow 'neck' region anterior to the genital pore and a long striated 'tail' region in the hindbody which is devoid of organs, such that the bulk of the uterus is restricted to the anterior half of the hindbody and the testis is located more anteriorly. F. gibsoni differs from F. glabra in having a longer forebody, smaller sucker width ratio and larger eggs, and from F. mugilis in having a smaller sucker width ratio, a hermaphroditic sac much longer than the ventral sucker and smaller eggs. The three Mediterranean forms, D. perpusilla, D. contracta and F. gibsoni are further distinguished by multivariate analyses. Keys to the species of both genera are presented. PMID- 19189231 TI - A revision of the haploporinae nicoll, 1914 (digenea: haploporidae) from mullets (mugilidae): two new haploporine genera and a key to the genera of the subfamily. AB - Two new haploporine genera are established for parasites of mullets. Ragaia n. g. is erected for R. lizae n. sp. from Liza ramado in the Ebro Delta on the Mediterranean Coast of Spain. This new genus is distinguished by the unique combination of the following characters: a strongly muscular ventral sucker which is twice as large as the oral sucker; a large, muscular hermaphroditic sac similar in length to the ventral sucker; a saccular, thick-walled internal seminal vesicle which is larger than the external seminal vesicle; and the ovary and vitellarium located rather close to the posterior extremity. Pseudodicrogaster n. g. is erected to accommodate Dicrogaster japonica Machida, 1996, as P. japonica (Machida, 1996) n. comb., a parasite of Mugil cephalus L. off Fukaura, Japan. This genus is recognised on the basis of: the tubular condition of both the internal and external seminal vesicles, the latter being much shorter than the former; the sucker ratio; the massive pyriform hermaphroditic sac; the location of the testis; and the presence of two eye-spots in developed miracidia. A key to the nine recognised genera of the Haploporinae is presented. PMID- 19189232 TI - Two new species of Anthobothrium van Beneden, 1850 (tetraphyllidea: phyllobothriidae) from carcharhinid sharks, with a redescription of Anthobothrium laciniatum Linton, 1890. AB - Anthobothrium laciniatum Linton, 1890 is redescribed based on specimens taken from the dusky shark Carcharhinus obscurus (Lesueur) collected from the Northwestern Atlantic Ocean, and a neotype is designated. A. laciniatum differs from A. cornucopia van Beneden, 1850, A. altavelae Euzet & Ben Hassine, 2002, A. lesteri Williams, Burt & Caira, 2004 and A. spinosum Subhapradha, 1955 in total length. It further differs from A. cornucopia, A. altavelae and A. spinosum in proglottid number, and differs from A. galeorhini Suriano, 2002, A. cornucopia, and A. spinosum in testis number. A. lyndoni n. sp. is described from the sandbar shark C. plumbeus (Nardo). This new species differs from A. laciniatum in ovarian width and from A. cornucopia, A. altavelae, A. galeorhini and A. spinosum in the total number of proglottids. It further differs from A. cornucopia, A. galeorhini, and A. spinosum in total length, and from A. cornucopia and A. galeorhini in the number of testes. A. lyndoni n. sp. differs from A. lesteri in bothridial muscle and ovarian morphology. Anthobothrium caseyi n. sp. is described from Prionace glauca (Linnaeus). This new species differs conspicuously from the other six species of Anthobothrium van Beneden, 1850 (sensu stricto) in the shape of its proglottid laciniations. The taxonomic status of 43 species that have been associated with Anthobothrium is addressed. Taxonomic actions regarding Anthobothrium during the past century have resulted in a polyphyletic taxon. PMID- 19189233 TI - A review of spawning induction, larviculture, and juvenile rearing of the fat snook, Centropomus parallelus. AB - The fat snook, Centropomus parallelus, is a commercially valuable marine fish species with potential for aquaculture. This paper describes the development of technology for mass production of fat snook juveniles at the Experimental Fish Hatchery of the Universidade Federal de Santa Catarina, focusing on research about reproduction, larviculture, and juvenile rearing. Induced spawning of wild fat snook was first achieved in 1991 with a single injection of human chorionic gonadotropin (hCG). There was a substantial increase in egg quality when broodstock was conditioned in maturation rooms and induced to spawn. Different dosages of luteinizing hormone-releasing hormone analogue (LHRHa) with saline injection and colesterol implant were also tested. As fat snook exhibits group synchronous oocyte development, females could be induced to spawn (with 35-50 mug kg(-1) of LHRHa) once a month, resulting in up to four consecutive spawnings. Results of larval culture were highly variable at the beginning; survival rates were frequently around 1% until the juvenile stage. Several experiments were conducted to evaluate the effect of environmental factors and feeding quality on survival and growth. With the improvement of the spawning induction technique and better larviculture practices, survival rates increased to 10-30%. Studies on the particular requirements of juveniles in terms of stocking density, feeding, nutrition, and environmental factors were also performed in order to improve growth rates and feed utilization. The present study demonstrates the feasibility of mass production of fat snook juveniles. However, further research is needed to develop cost-effective grow-out technology. PMID- 19189234 TI - Fishes of southern South America: a story driven by temperature. AB - The latitudinal extension of southern South America imposes a thermal gradient that affects the structure of marine and freshwater fish assemblages and the biology of the species through direct exposure to the temperature gradients or by means of a web of historical and ecological relationships. We have reviewed biological and ecological data of marine and freshwater fishes from the southern Neotropics, including Patagonia, and report several examples of dependence on temperature, from glacial times to today's climate change. We were able to identify historic and present effects on the diversity of fish assemblages, isolation, southern limits for the distribution of species, and morphological variation among populations. There is a wide range of characteristics that exemplify an adaptation to low temperatures, including biochemical peculiarities, physiological adjustments, and alternative life history patterns, and these appear in both freshwater and marine, and native and exotic fishes. The consequences of stable temperature regimes in both the ocean and thermal streams deserve special mention as these shape specialists under conditions of low selective pressure. At present, habitat use and interactions among species are being subject to changes as consequences of water temperature, and some of these are already evident in the northern and southern hemispheres. PMID- 19189235 TI - Patterns of commercial fish landings in the Loreto region (Peruvian Amazon) between 1984 and 2006. AB - Patterns of commercial fish catches over the period from 1984 to 2006 were studied in the Loreto region and in Iquitos, which is the most important town of the region and the principal fish marketplace of the Peruvian Amazon. Despite important inter-annual variations, the overall fish landings have significantly increased in the region during this period. The same three species dominated the catches during the whole period (Prochilodus nigricans, Potamorhina altamazonica and Psectrogaster amazonica), making up about 62% of the catches. However, the number of species exploited by commercial fisheries increased considerably during the 22 years of this study (from about 21 species in 1984 to over 65 in 2006), although part of the difference may be accounted for by a better identification of individual species nowadays. At the same time, the large high-valued species, such as Arapaima gigas, Colossoma macropomum and Brachyplatystoma rousseauxii, declined significantly and were replaced by smaller, short-lived and lower-valued species. Catches of the silver Arahuana (Osteoglossum bicirrhosum) also declined significantly during the studied period, strengthening recent warnings about the species' conservation status (Moreau and Coomes, Oryx 40:152-160, 2006). The relative proportions of the trophic groups (detritivores, omnivores and piscivores) remained relatively constant over the study period, but there were significant changes in the relative abundances of the species groups. The proportion of the dominant group, the Characiformes, which averaged about 81% of the catches, increased between 1984 and 2006, whereas the proportion of the Siluriformes and Perciformes remained constant. On the other hand, the proportion of Osteoglossiformes, represented only by two well known species (Arapaima gigas and Osteoglossum bicirrhosum), declined sharply during the same period. Important differences were observed between the landings of Iquitos and the landing of the whole Loreto region, indicating that conclusions drawn from the study of the Iquitos landings cannot be extrapolated to the whole landings of the Loreto region. The most important difference was the decreasing fish landings in Iquitos, whereas the total landings increased in the Loreto region at the same time. Potential causes of this phenomenon are discussed. Decreasing fish abundance around Iquitos (because of a higher fishing pressure) and a behavioural adaptation of fishermen to better law enforcement in Iquitos are likely explanations to be further investigated. PMID- 19189236 TI - Nutrition and feeding research in the spotted rose snapper (Lutjanus guttatus) and bullseye puffer (Sphoeroides annulatus), new species for marine aquaculture. AB - The spotted rose snapper (Lutjanus guttatus) and bullseye puffer (Sphoeroides annulatus) are fish species from the tropical Eastern Pacific for which controlled production of larvae and juveniles has been accomplished in recent years. Diverse topics relating to their biology and aquaculture production are currently under study, in particular the nutrition and feeding aspects required to formulate practical feeds and rearing protocols. Improvements in larval growth and survival are possible by feeding live food organisms with natural or enhanced essential fatty acids content and highly digestible artificial microdiets. The ontogeny of the digestive tract and the expression and activity of digestive enzymes have been described for S. annulatus larvae. The effect of various protein and lipid levels on growth and feed utilization has been studied in juvenile and on-growing fish. Both species have carnivorous feeding habits and require high levels of protein in their diets, from 40% to 45% (dry weight) in spotted rose snapper and above 50% in bullseye puffer, with the younger stages requiring the highest protein levels. Encouraging results have been obtained in feeding experiments with different sources of dietary protein from animal and plant origin to evaluate their suitability as feed ingredients in practical diets. Optimization of fish culture practices through feeding management has also been investigated. Trials with various fish densities and feeding frequencies in intensive culture systems are providing information to improve feed utilization and growth in on-growing fish. Further research is underway to evaluate factors in broodstock nutrition which have an impact on egg and larval quality, and into the use of various commercially available oil sources in on-growing diets. In this paper, the results on nutrition and feeding research with both species are reviewed and research needs to support their commercial production in the region are discussed. PMID- 19189237 TI - Effects of light and temperature conditions on the expression of GnRH and GtH genes and levels of plasma steroids in Odontesthes bonariensis females. AB - In this study we examined the endocrine mediation between environmental factors (temperature and photoperiod) and the brain-pituitary-gonadal axis in females of pejerrey Odontesthes bonariensis. Changes in the expression of brain gonadotropin releasing hormones (GnRHs) and gonadotropin (GtH) subunit [follicle stimulating beta (FSH-beta), luteinizing hormone-beta (LH-beta), glycoprotein hormone-alpha (GPH-alpha)] genes, plasma gonadal steroids [estradiol (E(2)) and testosterone (T)], gonadal histology, and gonadosomatic index (GSI) in adult females exposed to combinations of short-day (8 h) or long-day (16 h) photoperiods and low (12 degrees C) or high (20 degrees C) temperatures after winter conditions (8 h light, 12 degrees C) were analyzed. Pejerrey females kept under the short photoperiod had low GSIs, and their ovaries contained only previtellogenic oocytes regardless of the experimental temperature. In contrast, females exposed to the long photoperiod had high GSIs and ovaries with vitellogenic oocytes at both temperatures. These fish also showed a significantly higher expression of sGnRH, pjGnRH, cGnRH-II (the three different GnRH variants found to date in the pejerrey brain), FSH-beta, LH-beta and GPH-alpha genes and plasma E(2 )levels than those at the shorter photoperiod. No significant changes were observed in plasma T levels. Based on these results, we concluded that the increase in day length but not that of temperature triggers the maturation of pejerrey females after the winter period of gonadal rest and that this occurs by an integrated stimulation of the various components of the brain-pituitary-gonad axis. PMID- 19189238 TI - Patterns of oocyte development in natural habitat and captive Salminus hilarii Valenciennes, 1850 (Teleostei: Characidae). AB - Fecundity and oocyte development in Salminus hilarii female brood stock were analyzed with the aim of investigating the impact of migration impediment on oogenesis. Histological analyses of the ovaries were performed in adult females caught in two different environments--the Tiete River (natural) and captivity- and the gonadossomatic index, oocyte diameter and fecundity determined. Five germ cell development stages (oogonium, perinucleolar, cortical alveoli, vitellogenic, ripe) and two other structures (postovulatory follicles and atretic oocytes) were observed in females caught in the river. Captive animals lacked the ripe oocytes and postovulatory follicles and had a relatively higher number of atretic oocytes. Females in captivity are known to produce larger oocytes, and they release fewer eggs in each spawn (absolute fecundity) when compared with animals that are able to migrate. Our results suggest that the Tiete River is undergoing alterations which are being reflected in the reproductive performance of S. hilarii, mainly due to the presence of atretic oocytes in females caught in the river. The lack of postovulatory follicles and ripe oocytes in captive animals reveals that migratory impediment negatively impacts final oocyte maturation. However, the stage of maturation reached is adequate for ovulation induction with hormone manipulation. PMID- 19189239 TI - Gender determination in the Paiche or Pirarucu (Arapaima gigas) using plasma vitellogenin, 17beta-estradiol, and 11-ketotestosterone levels. AB - Arapaima gigas is an air-breathing giant fish of Amazonian rivers. Given its great economic and cultural importance, the aquaculture development of this species represents an evident solution to face the decline of wild populations. In captivity, reproduction occurs generally in large earthen ponds where stocks of a few tens of brooders are maintained together at the beginning of the rainy season (December-March in the Peruvian Amazon). Fry production relies on the spontaneous formation of male and female pairs, which build a nest, delimit a territory and guard the offspring for at least 20 days from other congeners and predators. However, as sex determination of A. gigas is not possible by morphological criteria, it is very difficult to optimize reproduction conditions and fry production in each pond, which seriously hampers the culture of this species. This situation prompted us to develop sexing methodologies based on (1) the detection of female specific plasma Vitellogenin (Vtg) using an enzyme immuno assay (EIA), and (2) the determination of plasma 17beta-estradiol and 11 ketotestosterone levels for immature specimens. The Vtg purification was performed by electro-elution after polyacrilamide gel electrophoresis (PAGE) from plasma of 17beta-estradiol treated A. gigas juveniles. Two different Vtg molecules were isolated, (Vtg(1) and Vtg(2)) with 184 and 112 kDa apparent molecular masses, respectively, and two antibodies were raised in rabbits for each Vtg molecule. Adult fish were 100% accurately sexed by Vtg EIA, while 100% of immature fish and 95% of adults were accurately sexed by 17beta-Estradiol and 11-Ketestosterone ratios. We also observed different color pattern development in male and female adult fish (6-year-olds) around the reproductive period. PMID- 19189240 TI - Sperm quality and cryopreservation of Brazilian freshwater fish species: a review. AB - The Brazilian freshwater fish diversity is the richest in the world. Only 0.7% of all Brazilian species have had any aspect of their sperm biology addressed up to this date. The majority of the fish species described in this review migrate during the spawning season (a phenomenon known as piracema). Urbanization, pollution, hydroelectric dams and deforestation are some of the causes of stock depletion or even local extinction of some of these species. The knowledge concerning sperm quality and minimum sperm:egg ratio is important to maximize the use of males without reducing hatching rates. Furthermore, sperm cryopreservation and gene banking can guarantee the conservation of genetic diversity and development of adequate breeding programs of native fish species. In this review, we present and evaluate the existing information on Brazilian fish species that have been subject to sperm quality and cryopreservation studies. The following parameters were evaluated: volume of extractable sperm, sperm motility, sperm concentration, freezing media, freezing methods, and post-thaw sperm quality. Although the existing protocols yield relatively high post-thaw motility and fertilization rates, the use of cryopreserved sperm in routine hatchery production is still limited in Brazil. PMID- 19189241 TI - Water pH and Prochilodus lineatus larvae survival. AB - The present study aimed to determine the influence of water pH on survival of curimbata, Prochilodus lineatus, larvae. Forty-five 2-l glass aquaria with artificial aeration and no water exchange were stocked at a density of 10 larvae l(-1). The pH values used as treatments were 3.7 +/- 0.0, 4.0 +/- 0.0, 4.4 +/- 0.2, 4.6 +/- 0.2, 4.8 +/- 0.2, 5.1 +/- 0.2, 5.3 +/- 0.2, 5.6 +/- 0.2, 7.2 +/- 0.2, 8.7 +/- 0.4, 9.0 +/- 0.4, 9.2 +/- 0.4, 9.4 +/- 0.6, 9.7 +/- 0.5 and 10.0 +/- 0.5 in a completely random experimental design with three repetitions. Water pH was maintained by the addition of NaOH or H(2)SO(4) solutions. After 72 h of experiment, no survival was registered at pH below 4.6, 1.5% survival at pH 4.6, and about 50% survival at pH 9.4 and above. Survival rates between 70 and 80% were registered at pH 4.8-5.6 and at pH 7.2, whereas over 90% survival was registered at pH between 8.7 and 9.2. PMID- 19189243 TI - Energy density of sub-Antarctic fishes from the Beagle Channel. AB - The energy density (ED) of nine species of sub-Antarctic fishes was estimated by calorimetry. The fish, seven notothenioids, one atherinopsid and one galaxiid, represents some of the more abundant species in the ichthyofauna of the Beagle Channel. Principal-components analysis (PCA) of the ED of the different organs/tissues indicated that PC(1) and PC(2) accounted for 87% of the variability. Separation along PC(1) corresponded to differences in muscle and liver energy densities whereas separation along PC(2) corresponded to differences in the ED of the gonads. Differences between species were significant except for P. sima. Inclusion of the gonadosomatic index (GSI) as an explanatory variable enabled us to establish the existence of energy transfer from muscle and liver to the gonads in ripe P. tessellata females. Total ED values varied between 4.21 and 6.26 kJ g(-1), the pelagic Odontesthes sp. being the species with the highest ED. A significant relationship between ED and muscle dry weight (DW(M)) was found for all the species except P. tessellata. These data are the first direct estimates of ED of fishes from the Beagle Channel. PMID- 19189242 TI - mRNA expression of GnRH variants and receptors in the brain, pituitary and ovaries of pejerrey (Odontesthes bonariensis) in relation to the reproductive status. AB - The present study examined the differential mRNA expression levels of three forms of GnRH (sGnRH, pjGnRH and cGnRH-II) and two forms of GnRH receptor (pjGnRH-R I and pjGnRH-R II) in the brain, pituitary, and ovaries of pejerrey in relation to the reproductive status. The analysis revealed the presence of significant amounts of mRNA of the three GnRH forms while the ovaries showed only two (sGnRH and pjGnRH). The GnRH receptor II was found ubiquitously in the brain, pituitary, and ovaries while the form I was detected only in the brain. The levels of pjGnRH mRNA in the brain and pjGnRH-R II in the pituitary gland varied in correlation with the ovarian condition. However, brain sGnRH and pjGnRH-R I mRNA levels reached a maximum during early stages of ovarian development. In contrast, the brain levels of cGnRH-II mRNA showed no variation. The present study also shows a good correlation of ovarian sGnRH and pjGnRH-R II mRNA levels with the reproductive condition, suggesting that these molecules are may be involved in the regulation of pejerrey ovarian function. PMID- 19189244 TI - Sema4C expression in neural stem/progenitor cells and in adult neurogenesis induced by cerebral ischemia. AB - Sema4C is a transmembrane protein that belongs to axon guidance molecules of semaphorin family. Previous reports have shown that Sema4C could interact with postsynaptic protein PSD95, etc, but the expression and the role of Sema4C in neurogenesis remains unknown. In this study, whole-mount in situ hybridization result showed that Sema4C was expressed abundantly in the areas of lateral ventricle, the striatum, the wall of midbrain, and the pons/midbrain junction of E11.5 embryos brain. Neural stem/progenitor cells (NSPs) obtained from E13.5 embryonic rat midbrain are also positive for Sema4C immunoreactivity. Sema4C expression was dramatically downregulated during induction of NSP differentiation. In order to confirm the involvement of Sema4C in neurogenesis, we used the rat global cerebral ischemia model to make adult neurogenesis in vivo. The robust proliferative NSPs were monitored by labeling with bromodeoxyuridine (BrdU) within the subventricular zone and dentate gyrus that continues for at least 2 weeks. Immunohistochemistry and Western blot analysis showed that Sema4C expression was dramatically upregulated during neurogenesis after cerebral ischemia-perfusion injury. Double immunostaining and stereologic counting analysis indicated that a high proportion of BrdU-positive proliferative cells were Nestin-positive NSPs, and also, Sema4C was highly expressed in these proliferative populations at specific stages after ischemic injury. These observations provide the evidence to support a putative role of Sema4C during neurogenesis both in vivo and in vitro. PMID- 19189246 TI - Exploratory study on the clinical and mycological effectiveness of a herbal medicinal product from Solanum chrysotrichum in patients with Candida yeast associated vaginal infection. AB - Mexican traditional medicine uses Solanum chrysotrichum to treat fungi-associated dermal and mucosal illness; its methanolic extract is active against dermatophytes and yeasts. Different spirostanic saponins (SC-2-SC-6) were identified as the active molecules; SC-2 was the most active in demonstrating a fungicidal effect against Candida albicans and non-albicans strains. The aim of the present study was to compare the clinical (elimination of signs and symptoms) and mycological effectiveness (negative mycological studies) of an S. chrysotrichum herbal medicinal product (Sc-hmp), standardized in 1.89 mg of SC-2, against ketoconazole (400 mg) in the topical treatment of cervical and/or vaginal infection by Candida. Both treatments (vaginal suppositories) were administered daily during 7 continuous nights. The study included 101 women (49 in the experimental group) with a confirmed clinical condition and positive mycological studies (direct examination and/or culture) of Candida infection. Basal conditions did not show differences between the groups; a moderate clinical picture was present in 62% of the cases, direct examination was positive in 69%, and the culture was positive with C. albicans predominating (65%). At the end of the administration period, both treatments demonstrated 100% tolerability, and clinical cure in 57.14% of S. chrysotrichum-treated cases and in 72.5% of ketoconazole-treated cases (p = 0.16), as well as 62.8% and 97.5% of mycological effectiveness, respectively (p = 0.0 001). We conclude that, at the doses used, Sc-hmp exhibits the same clinical effectiveness as ketoconazole, but with lower percentages of mycological eradication. Additional clinical studies with Sc-hmp are necessary, with increasing doses of SC-2, for improving the clinical and mycological effectiveness. PMID- 19189245 TI - New constituents with iNOS inhibitory activity from mycelium of Antrodia camphorata. AB - In continuing our investigation on the bioactive constituents of mycelium of Antrodia camphorata, antroquinonol B (1), 4-acetyl-antroquinonol B (2), 2,3 (methylenedioxy)-6-methylbenzene-1,4-diol (3) and 2,4-dimethoxy-6-methylbenzene 1,3-diol (4) along with antrodin D (5) were isolated by the guidance of an inducible nitric oxide synthase (iNOS) inhibitory assay and identified on the basis of their spectroscopic analysis. The effect of these compounds on the inhibition of NO production in lipopolysaccharide (LPS)-activated murine macrophages was further evaluated. Compounds 4 and 5 significantly inhibited NO production without any cytotoxicity, the IC(50) values being 32.2 +/- 0.1 and 26.3 +/- 1.6 microg/mL, respectively. Compounds 1 and 2 possessed greater effects on NO inhibition, with IC(50) values of 16.2 +/- 0.8 and 14.7 +/- 2.8 microg/mL, respectively, but displayed cytotoxicity at considerably higher concentrations. Compound 3 showed the lowest percent cell viability of 45.5 +/- 1.8 % as observed in treated cells at a concentration of 16.8 microg/mL. PMID- 19189247 TI - Authentication of Panax ginseng from its adulterants by PCR-RFLP and ARMS. AB - As a widely used and expensive herbal medicine, Panax ginseng has many adulterants in the commercial market. PCR-restriction fragment length polymorphism (PCR-RFLP) and amplification refractory mutation system (ARMS) based on 5S rDNA sequence analysis were applied to identify two common adulterants of P. ginseng. The sizes of 5S rRNA gene non-transcribed spacers (NTS) sequences in P. ginseng and its adulterants were determined, ranging from 143 to 424 bp. The PCR product of P. ginseng only could be digested among the tested specimens because of its specific SpeI restriction site found in the 5S rDNA sequence. In addition, P. ginseng was successfully identified from compound medicinal preparations and from the Single-Taste medicines. These results suggest that the methods are able to authenticate P. ginseng. PMID- 19189248 TI - [Wrist joint arthroplasty: results after 41 prostheses]. AB - PURPOSE: The advantage of wrist arthroplasty remains controversial, primarily due to the high complication rate. For this reason it seems sensible to monitor the results of different types of prostheses even with small numbers of cases. We were particularly interested to see if wrist joint arthroplasty is a useful alternative for patients with rheumatoid arthritis, and which of the types we used shows the best results. PATIENTS AND METHODS: In our hospital, 41 wrist joint prostheses (15 Meuli, 16 BIAX and 10 Universal 2) were implanted in 36 patients from 1992 until 2005 (follow-up 1 to 14 years, mean 5.3 years). 33 patients had rheumatic destruction of the wrist, two had osteoarthritis following fracture of the scaphoid, and one pseudarthrosis after failed arthroplasty and arthrodesis for Kienbock's disease. Mean age was 54 years, ranging from 34 to 73 years. 14 patients had had surgery on this wrist before. The patients were sent a questionnaire including the DASH score, and a clinical evaluation and X-rays were performed. RESULTS: 33 patients with 38 wrist arthroplasties answered the questionnaire, 34 wrist joint prosthesis of 29 patients could be evaluated. DISCUSSION: 6 prostheses had to be removed because of complications (3 arthrodeses were performed after removal, 3 prostheses were exchanged). There were 4 dislocations (3 times with the Meuli type, once with the BIAX type). There was one case of CRPS type I. But subjectively, in answering our questionnaire, 31 of 38 patients claimed to be very satisfied or satisfied with the result of the operation, only 6 were less satisfied or not satisfied at all. An improvement of pain was found by all but one patient. An increase in strength or range of movement was found more rarely. The mean postoperative DASH score was 61 points. Mean wrist joint mobility was 50 degrees for extension/flexion, and 20 degrees for radial- and ulnar abduction. CONCLUSION: The result of total wrist joint arthroplasty depends very much on a careful patient selection. A preoperative bony malposition of the wrist and a tendon dysfunction seem to be responsible for a bad result. High expectations with regard to range of movement and strength should be avoided. A good reduction of pain can be achieved but the risk of complication is still higher than in arthrodesis. PMID- 19189249 TI - [Suppression of reproductive cyclicity by active immunization against GnRH in the adult ewe]. AB - The objective of the present study was to evaluate the effect of an anti-GnRH vaccine on cycling activity in the adult ewe during the breeding season. For the experiments 22 cycling White Alpine sheep, aged between 2 and 4 years, were randomly divided into a treatment and control group of 11 animals, each. Sheep of the treatment group were immunized twice at an interval of 4 weeks with 2 ml (400microg GnRH-protein conjugate) of Improvac (Pfizer Animal Health, Australia) subcutanously in the neck. Sheep of the control group received the same amount of saline solution. Blood progesterone concentrations were measured weekly from 3 weeks before to 32 weeks after first immunization and anti-GnRH titers were determined monthly. All vaccinated ewes ceased cycling within 2 - 8 weeks after first immunization. Plasma progesterone was suppressed for a minimum of 12 weeks (2 ewes) with individual variation of 14 (1 ewe), 25 (1 ewe) and more than 25 weeks (7 ewes). Four animals resumed cyclicity while 7 animals remained suppressed until the end of the study. Antibody titers peaked one month after the booster injection and thereafter continuously dropped until the end of the study (8 months after first immunization) to values between 10.9 and 40.8 % binding. From our results it can be concluded that two vaccinations with Improvac 4 weeks apart suppress cycling activity in adult ewes for at least 12 weeks. The inhibitory effect on ovarian activity, however, varies individually and may last more than 31 weeks. PMID- 19189250 TI - Changes in body weight, hematology and serum chemistry in captive plains viscachas (Lagostomus maximus) with presumptive diabetes type II after a diet change. AB - We report body weights (BW) and blood and serum analyses for 6 fully mature and 8 not-yet-mature captive plains viscachas before and 3, 6 and 9 months after switching from a low-fiber, high-energy diet to a high-fiber, low-energy diet. Initially, body weights, serum glucose, fructosamine and cholesterol levels were above the reference range in the fully mature animals. Furthermore, 4 of these animals had bilateral cataracts. After the diet change, these parameters dropped into the reference range. However, 9 months later, a slightly increased BW became evident again. The findings are consistent with a type II diabetes mellitus and underline the importance of dietary prevention. PMID- 19189251 TI - Multiple organ dysfunction syndrome in a dog with Klebsiella pneumoniae septicemia. AB - Multiple organ dysfunction syndrome is defined as the presence of altered organ function in an acutely ill patient such that homeostasis could not be mantained without intervention. It is reported that mortality rate in humans with 4 organ systems failing reaches nearly 100%, while there are few publications documenting this syndrome in dogs. To the author's knowledge, this is the first report that describes the clinical manifestations and the favourable outcome to intensive medical care in a dog with Klebsiella pneumoniae septicemia and multiple organ dysfunction with 6 organ systems failing. Derangement of cardiovascular, respiratory, gastrointestinal, pancreatic, renal and coagulation system developed. This dog manifested reversible myocardial depression that is a common complication of sepsis in people but it is rarely reported in dogs. PMID- 19189253 TI - [Tracheal collapse with dyspnea in 2 calves with multiple rib fractures]. PMID- 19189252 TI - [Zinc intoxication in a Yorkshire Terrier due to Euro cent ingestion]. AB - A 5-year old, intact male Yorkshire Terrier was presented due to lethargy, vomiting and diarrhea. Clinical signs included shock, icterus and a painful abdomen. Laboratory examination revealed a severe intravascular hemolytic anemia, a thrombocytopenia and a leukocytosis. The radiographic examination revealed the presence of metallic foreign bodies in the stomach and in the intestine. After stabilisation of the patient with crystalloids, packed red blood cells, ranitidine, metamizole, amoxicilline/clavulanic acid and marbofloxacine, surgery was performed and 5 coins (10 cent, 5 cent, 2 cent) were removed via esophagus and via enterotomy. In the course of disease the dog developed acute renal failure and pancreatitis. The zinc concentration in the serum was 2200 microg/dl, which was 30 times higher compared to a healthy control dog. After 21 days the dog was discharged from the hospital. On day 28 all laboratory values were within the reference range. PMID- 19189255 TI - [The Swiss social insurance law]. PMID- 19189256 TI - [Attention! matter for the boss: the art of delegation]. PMID- 19189257 TI - [Continuing education day for TPA in western Switzerland]. PMID- 19189267 TI - N-(ureidoethyl)amides of cyclic enkephalin analogs. AB - Novel N-(ureidoethyl)amides of cyclic enkephalin analogs have been synthesized. The p-nitrophenyl carbamate of 1-Boc-1,2-diaminoethane was coupled with 4 methylbenzhydrylamine (MBHA) resin. The Boc group was removed by treatment with HCl/dioxane, and the peptide chain was assembled using Boc strategy. For deprotection of amino function, HCl/dioxane was used. D-Lys or D-Orn were incorporated in position 2, and the side chains of Lys, Orn, Dab, or Dap in position 5 were protected with Fmoc group. Side chain protection was removed by treatment with 55% piperidine in DMF, and cyclization was achieved by treatment with bis-(4-nitrophenyl)carbonate to form a urea bridge. The peptide was cleaved from the resin by treatment with 45% TFA in DCM. The peptides were tested in the guinea-pig ileum (GPI) and mouse vas deferens (MVD) assays. Divers opioid activities were observed, depending on the size of the ring. In comparison with [Leu(5)]enkephalin, all peptides were more active in the GPI assay (between 125 and 12 times), and some of them were also more potent in the MVD assay. The conformational propensities of each peptide were determined using the EDMC method in conjunction with NMR experiments. This approach allows treating the dynamical behavior of small peptides properly. The results were compared with those obtained previously for corresponding nonsubstituted amides and are in agreement with the biologically active conformation proposed by us earlier. PMID- 19189268 TI - Synthesis of hepcidin derivatives in order to develop standards for immune adsorption method. AB - MeCN, acetonitrile; ECL, enhanced chemiluminescence; EDT, 1,2-ethanedithiole; HEPC12-A, rabbit anti-human hepcidin IgG, affinity purified; HEPC13-A, rabbit anti-mouse/human hepcidin IgG, affinity purified; HEPC61-P, human hepcidin-25 control/blocking synthetic peptide; HRP, horseradish peroxidase; IL-6, interleukin-6; KLH, keyhole limpet hemocyanin; LEAP, liver-expressed antimicrobial peptide; NEM, N-ethylmaleimide; NMP, N-methyl-pirrolidone; PBS, phosphate buffered saline; PVDF, polyvinylidene difluoride; SELDI-TOF-MS, surface enhanced laser desorption ionization-time-of-flight mass spectrometry; TMB, tetramethylbenzidin; TNF-alpha, tumor necrosis factor-alpha. PMID- 19189269 TI - Complexes of heterocyclic aza-derivatives of phytoalexin from cotton plant gossypol with alkali metal cations and perchlorate anion studied by ESI mass spectrometric method in the positive and negative modes. PMID- 19189270 TI - Thermodynamics of partitioning of substance P in isotropic bicelles. AB - The temperature dependence of the partition of a neuropeptide, substance P (SP), in isotropic (q = 0.5) bicelles was investigated by using pulsed field gradient NMR diffusion technique. The partition coefficient decreases as the temperature is increased from 295 to 325 K, indicating a favorable (negative) enthalpy change upon partitioning of the peptide. Thermodynamic analysis of the data shows that the partitioning of SP at 300 K is driven by the enthalpic term (DeltaH) with the value of - 4.03 kcal mol(-1), while it is opposed by the entropic term (-TDeltaS) by approximately 1.28 kcal mol(-1) with a small negative change in heat capacity (DeltaC(p)). The enthalpy-driven process for the partition of SP in bicelles is the same as in dodecylphosphocholine (DPC) micelles, however, the negative entropy change in bicelles of flat bilayer surface is in sharp contrast with the positive entropy change in DPC micelles of highly curved surface, indicating that the curvature of the membrane surface might play a significant role in the partitioning of peptides. PMID- 19189271 TI - Antiestrogenic and anticancer activities of peptides derived from the active site of alpha-fetoprotein. AB - Cyclo[EKTOVNOGN] (AFPep), a cyclic 9-amino acid peptide derived from the active site of alpha-fetoprotein, has been shown to prevent carcinogen-induced mammary cancer in rats and inhibit the growth of ER(+) human breast cancer xenografts in mice. Recently, studies using replica exchange molecular dynamics predicted that the TOVN region of AFPep might form a dynamically stable putative Type I beta turn, and thus be biologically active without additional amino acids. The studies presented in this paper were performed to determine whether TOVN and other small analogs of AFPep would inhibit estrogen-stimulated cancer growth and exhibit a broad effective-dose range. These peptides contained nine or fewer amino acids, and were designed to bracket or include the putative pharmacophoric region (TOVN) of AFPep. Biological activities of these peptides were evaluated using an immature mouse uterine growth inhibition assay, a T47D breast cancer cell proliferation assay, and an MCF-7 breast cancer xenograft assay. TOVN had very weak antiestrogenic activity in comparison to AFPep's activity, whereas TOVNO had antiestrogenic and anticancer activities similar to AFPep. OVNO, which does not form a putative Type I beta-turn, had virtually no antiestrogenic and anticancer activities. A putative proteolytic cleavage product of AFPep, TOVNOGNEK, significantly inhibited E(2)-stimulated growth in vivo and in vitro over a wider dose range than AFPep or TOVNO. We conclude that TOVNO has anticancer potential, that TOVNOGNEK is as effective as AFPep in suppressing growth of human breast cancer cells, and that it does so over a broader effective-dose range. PMID- 19189272 TI - Investigation of the synthetic route to pepstatin analogues by SPPS using O protected and O-unprotected statine as building blocks. AB - The synthetic route to pepstatin derivatives by a solid phase peptide synthesis using either O-protected or O-unprotected statine as a building block has been investigated. Statine was prepared according to a modified literature procedure, whereas protection of its 3-hydroxyl moiety using tert-butyldimethylsilylchloride (TBSCl) provided the novel O-TBS-protected statine building block. The O-tert butyldimethylsilyl (TBS)-protected statine approach provides an improved synthetic strategy for the preparation of statine-containing peptides as demonstrated by the synthesis of the pepstatin analogue iva-Val-Leu-Sta-Ala-Sta. PMID- 19189273 TI - Production and isotope labeling of antimicrobial peptides in Escherichia coli by means of a novel fusion partner that enables high-yield insoluble expression and fast purification. AB - A method is presented that allows efficient production of antimicrobial peptides in bacteria by means of fusion to the histone fold domain of the human transcription factor TAF12. This small fusion partner drives high-level expression of peptides and leads to their accumulation in an entirely insoluble form, thereby eliminating toxicity to the host. Using the antimicrobial peptide LAH4 as an example, we demonstrate that neither affinity purification of the TAF12 fusion protein nor initial solubilization of inclusion bodies in denaturing buffers is required. Instead, crude insoluble material from bacteria is directly dissolved in formic acid for immediate release of the peptide through chemical cleavage at a unique Asp-Pro site. This is followed by purification to homogeneity in a single chromatographic step. Because of the elevated expression levels of the histone fold domain and its small size (8 kDa), this straightforward purification scheme produces yields in excess of 10 mg active peptide per liter of culture. We demonstrate that TAF12 fusion allows expression of a wide range of antimicrobial peptides as well as efficient isotope labeling for NMR studies. PMID- 19189274 TI - The Distress Thermometer assessed in women at risk of developing hereditary breast cancer. AB - OBJECTIVES: The Distress Thermometer (DT) is a promising instrument to get insight into distress experienced by cancer patients. At our Family Cancer Clinic the DT, including an adapted problem list, was completed by 100 women at increased risk of developing hereditary breast cancer (mean age 45.2 years; SD: 10.5). Additionally, the women filled in either the Hospital Anxiety and Depression Scale as psychological component (n=48) or the somatic subscale of the Symptom Checklist-90 as somatic component (n=50) to identify associations with the DT-score. Further, the women filled in an evaluation form. RESULTS: The median score on the DT was 2 (range: 0-9). With regression analysis adjusted for age, the contribution of mood and somatic complaints, respectively, was investigated. The standardized regression coefficient for anxiety was 0.32 (ns), for depression 0.14 (ns) and for the somatic subscale 0.49 (p<0.001). The explained variance for anxiety and depression was 16%, and for somatic complaints 24%. The differences between the coefficients were not significant. Evaluation forms were returned by 73 women. In 50% of the cases, the physician had discussed the DT/problem list, which was appreciated by the majority of these women (80%). Sixty-two percent of the women would recommend the use of the DT for other patients. CONCLUSION: The use of the DT/problem list seems promising for the current population, and was appreciated by the majority of the women. As mood and somatic complaints did not differ significantly in explaining the experienced distress, other candidate factors need to be examined. PMID- 19189275 TI - Massage therapy reduces physical discomfort and improves mood disturbances in women with breast cancer. AB - BACKGROUND. A randomized controlled trial was conducted to investigate the efficacy of classical massage treatment in reducing breast cancer-related symptoms and in improving mood disturbances. METHODS. Women diagnosed with primary breast cancer were randomized into an intervention group and a control group. For a period of 5 weeks, the intervention group received bi-weekly 30-min classical massages in the back and head-neck areas. The control group received no additional treatment to their routine healthcare. To evaluate treatment efficacy, the following validated questionnaires were administrated at baseline (T1), at the end of the intervention (T2), and at a followup at 11 weeks (T3): the Short Form-8 Health Survey, the European Organization of Research and Treatment of Cancer quality of life questionnaire breast module (EORTC QLQ-BR23), the Giessen Complaints Inventory (GBB), and the Berlin Mood Questionnaire (BSF). RESULTS. Eighty-six eligible women (mean age: 59 years) were enrolled in the study. A significantly higher reduction of physical discomfort was found in the intervention group compared with the control group at T2 (p=0.001) and at T3 (p=0.038). A decrease in fatigue was also observed. Women in the intervention group reported significantly lower mood disturbances at T2 (p<0.01) but not at T3. The effect of treatment on mood disturbances was significantly higher if a patient was treated continuously by the same masseur. CONCLUSION. Classical massage seems to be an effective adjuvant treatment for reducing physical discomfort and fatigue, and improving mood disturbances in women with early stage breast cancer. PMID- 19189276 TI - The Max homodimeric b-HLH-LZ significantly interferes with the specific heterodimerization between the c-Myc and Max b-HLH-LZ in absence of DNA: a quantitative analysis. AB - Specific heterodimerization plays a crucial role in the regulation of the biology of the cell. For example, the specific heterodimerization between the b-HLH-LZ transcription factors c-Myc and Max is a prerequisite for c-Myc transcriptional activity that leads to cell growth, proliferation and tumorigenesis. On the other hand, the Mad proteins can compete with c-Myc for Max. The Mad/Max heterodimer antagonizes the effect of the c-Myc/Max heterodimer. In this contribution, we have focused on the specific heterodimerization between the b-HLH-LZ domains of c Myc and Max using CD and NMR. While the c-Myc and Max b-HLH-LZ domains are found to preferentially form a heterodimer; we demonstrate for the first time that a significant population of the Max homodimeric b-HLH-LZ can also form and hence interferes significantly with the specific heterodimerization. This indicates that the Max/Max homodimer can also interfere with c-Myc/Max functions, therefore adding to the complexity of the regulation of transcription by the Myc/Max/Mad network. The demonstration of the existence of the homodimeric population was made possible by the application of numerical routines that enable the simulation of composite spectroscopic signal (e.g. CD) as a function of temperature and total concentration of proteins. From a systems biology perspective, our routines may be of general interest as they offer the opportunity to treat many competing equilibriums in order to predict the probability of existence of protein complexes. PMID- 19189277 TI - Prediction of survival in Alzheimer's disease--the LASER-AD longitudinal study. AB - OBJECTIVES: Alzheimer's disease (AD) is associated with variable but shortened life expectancy. Knowing expected survival time may empower people with AD and their families, but clinicians currently have limited predictive information. Our objective was to identify determinants of survival in a cohort of people with mild to moderate AD and test these on a separate validation cohort. METHODS: We followed a representative cohort of 158 people for 42 months and identified independent determinants of shorter survival. From these we constructed the Survival in Alzheimer's Model (SAM), and tested this on a validation cohort. RESULTS: Baseline constructional apraxia, age and gait apraxia independently predicted shorter survival: about half of those scoring 2 on the SAM survived > or =3.5 years compared to 85% of those scoring 0. CONCLUSIONS: The SAM is a potentially useful tool for clinicians who previously had very limited specific and quantitative prognostic information to tell AD patients and carers. This model predicted survival from age, constructional and gait apraxia. This may be because constructional and gait apraxia are relatively free from educational or cultural bias and thus are better indicators of severe neuropathology than global cognitive tests. Alternatively, they may increase falls or immobility, or represent disease sub-types with worse prognoses. Oncology services are able to inform patients and their families about 5-year survival rates. This step towards such provision in AD is new and of potential importance to patients and their carers. PMID- 19189278 TI - IQ in late adolescence/early adulthood, risk factors in middle age, and later cancer mortality in men: the Vietnam Experience Study. AB - OBJECTIVES: (i) examine the relation, if any, of pre-morbid IQ scores at 20 years of age with the risk of later cancer mortality; and (ii) explore the role, if any, of potential mediating factors (e.g. smoking, obesity), assessed in middle age, in explaining the IQ-cancer relation. METHODS: Cohort study of 14, 491 male, Vietnam-era, former US army personnel with IQ test scores at around 20 years of age (1965-71), who participated in a risk factor survey at around age 38 years of age (1985-6), who were then followed up for mortality experience for 15 years. RESULTS: There were 176 cancer deaths during mortality surveillance. We found an inverse association of IQ with later mortality from all cancers combined (age adjusted HR(per one SD decrease in IQ); 95% confidence interval: 1.27; 1.10, 1.46) and smoking-related malignancies (1.37; 1.14, 1.64). There was some attenuation following control for mediating variables, particularly smoking and income, but the gradients generally held at conventional levels of statistical significance. CONCLUSIONS: Higher scores on pre-morbid IQ tests are associated with lower risk of later cancer morality. The strength of the relation was partially mediated by established risk factors. PMID- 19189279 TI - Personality traits and psychosocial stress: quality of life over 2 years following breast cancer diagnosis and psychological impact factors. AB - OBJECTIVE: The aim of this study was to investigate changes in the quality of life (QoL) and body image among breast cancer patients over 2 years and to examine different predictive factors for QoL 2 years after the primary operation. METHODS: A total of 203 women with a primary diagnosis of breast cancer completed the questionnaires 2 weeks and 6, 12, 18, and 24 months after surgery. Quality of Life Questionnaire (QLQ-C30), Breast Cancer Specific Quality of Life Questionnaire Module (QLQ-BR23), Questionnaire on Stress in Cancer Patients (QSC R23), Freiburg Personality Inventory (FPI-R), Life Orientation Test (LOT) were used as standardized measures. RESULTS: The overall QoL and most functional and symptom scales improved during the 2-year period. However, cognitive functioning, body image, and the three symptom scales of insomnia, constipation, and diarrhea did not change. Age was only capable of predicting physical functioning, whereas tumor size, axillary surgery, and adjuvant chemotherapy were not predictive of the long-term QoL functional scores. Initial distress was the most potent predictive factor for long-term QoL. Baseline functioning predicted functional QoL scores 2 years later. And higher scores for neuroticism were associated with a poorer QoL. However, optimism was not capable of predicting the QoL 2 years later. CONCLUSION: Screening measures should be implemented at the time when breast cancer is diagnosed, in order to identify psychologically vulnerable patients and offer them professional psycho-oncological help. PMID- 19189280 TI - Illness perceptions in Greek patients with cancer: a validation of the Revised Illness Perception Questionnaire. AB - OBJECTIVE: The Revised-Illness Perception Questionnaire (IPQ-R) assesses illness perceptions according to Leventhal's self-regulatory model. The aim of this paper is to present findings on the reliability and validity of the IPQ-R in a population of Greek cancer patients. METHODS: A total of 206 patients completed a Greek translation of the IPQ-R and the Greek version of the Beck Depression Inventory (BDI). The scale's reliability was investigated by examining its internal consistency (Cronbach's alpha) and its test-retest reliability. Structural validity was examined through factor analyses. Predictive validity was tested by regressing BDI scores on IPQ subscale scores. Inter-relationships between IPQ-R dimensions were also examined by computing Pearson's Correlation Coefficients. RESULTS: Cronbach's alpha showed satisfactory internal consistency for the IPQ-R subscales. Paired samples' t-test showed good test-retest reliability. Factor analysis of the IPQ-R items revealed that the Greek version reflects the structure of the original with the only difference being that the 'Consequences' and 'Emotional Representations' subscales loaded on one factor. Factor analysis of the causal dimension items revealed a different structure of Causal Representations than that of the original questionnaire yielding three main factors: Psychological Attributions, Behavioral, and External Factors. Multiple regression analyses showed that Consequences, Emotional Representations, Illness Identity, and Psychological Attributions were the best predictors for depression. CONCLUSIONS: Translation of the IPQ-R has good reliability and similar structure to that of the original. Difficulties to confirm the structure of Causal Representations may represent cultural differences in understanding illness causation. PMID- 19189281 TI - Structure elucidation of three diphenyl ether derivatives from the mangrove endophytic fungus SBE-14 from the South China Sea. AB - Two new natural products, tenelate A (1) and B (2), together with the known compound, tenellic acid C (3), were isolated from the mangrove endophytic fungus Talaromyces sp. (SBE-14), from the South China Sea. Their structures were elucidated by spectroscopic methods, mainly 1D and 2D NMR techniques. PMID- 19189282 TI - The latent structure of the Peritraumatic Dissociative Experiences Questionnaire. AB - The Peritraumatic Dissociative Experiences Questionnaire (PDEQ) is a widely used measure of peritraumatic dissociation, and is presumably a unidimensional construct. Two hundred forty-seven individuals admitted to five hospitals after traumatic injury were administered the Clinician Administered PTSD Scale, the Hospital Anxiety and Depression Scale, and the PDEQ. Factor analysis indicated that the PDEQ involved two factors containing four items each: one factor (altered awareness) indexes alterations in awareness and the other (derealization) reflects distortions in perceptions of the self and the world. Only the derealization factor was associated with acute stress, anxiety, and depression symptoms. Cross-validation with independent data provided only partial support for the 2-factor structure model. These data indicate that peritraumatic dissociation may involve two distinct constructs. PMID- 19189283 TI - (79)Br NMR spectroscopy as a practical tool for kinetic analysis. AB - (79)Br NMR spectroscopy has been used to monitor a series of reactions in which the bromide ion is produced, including the Menschutkin reaction of pyridine with a range of substituted benzyl bromides and a Heck coupling process. In cases where the process could also be monitored using (1)H NMR spectroscopy, the kinetic analyses using heteronuclear magnetic resonance spectroscopy were shown to be completely consistent. Both the utility of the process in following reactions which may be difficult to analyse using other techniques and the practical limitations associated with solvent choice are discussed. PMID- 19189284 TI - Comparison of toxicity and transfection efficiency of amphiphilic block copolymers and polycationic polymers in striated muscles. AB - BACKGROUND: Gene delivery using synthetic vectors is frequently based on cationic compounds such as polyethyleneimine (PEI). However, few data have been published on the ability of PEI to mediate transgene expression in muscle tissue. Besides cationic vectors, there is increasing interest focusing on amphiphilic copolymers as gene carriers into striated muscles, although their mechanism of action is unknown. METHODS: Plasmid DNA was associated with three different polymers: the cationic polyethyleneimine and two amphiphilic copolymers displaying few (tetronic 304) or no charges (pluronic L64). The resulting formulations were investigated by dynamic light scattering, laser doppler velocimetry, gel retardation assay and transmission electron microscopy. The toxicity and efficiency of the carriers were assessed in both skeletal and cardiac muscles. RESULTS: PEI efficiently condenses plasmids into small complexes displaying a positive electrophoretic mobility. However, these PEI/DNA complexes lead to severe side-effects in vivo. The association between amphiphilic copolymers and DNA leads to no or only partial condensation of plasmids. Moreover, amphiphilic polymers do not interact strongly with DNA and tetronic-based systems are destabilized with a decreasing pH. Those vectors also display a negative electrophoretic mobility. Thus, characteristics of amphiphilic polymer/DNA systems might be considered unfavourable for transfection. However, safe and rather efficient gene expression was obtained in skeletal muscles, even at low DNA doses, but not in the myocardium. CONCLUSIONS: The present study highlights the interest in amphiphilic carriers for promoting DNA transfection in vivo. Gaining new insights into the properties of these vectors should allow their optimization. PMID- 19189285 TI - Reduction of liver tumor necrosis factor-alpha expression by targeting delivery of antisense oligonucleotides into Kupffer cells protects rats from fulminant hepatitis. AB - BACKGROUND: Fulminant liver failure can cause extreme mortality due to the lack of effective and targeting therapeutics for the disease. Novel therapeutics using antisense technology require an efficient and safe delivery system with Kupffer cell targeting ability. METHODS: We explored the capacity of galactosylated low molecular weight chitosan (GLC) to efficiently mediate the antisense oligonucleotide (ASO) TJU-2755 into Kupffer cells, enhance the effect of the oligonucleotides on the suppression of tumor necrosis factor (TNF)-alpha and prolong the active time of the antisense drug in vivo. The protective and therapeutic effect of ASO/GLC in the animal model of D galactosamine/lipopolysaccharide-induced fulminant hepatitis was tested. RESULTS: ASOs delivered by GLC were concentrated in Kupffer cells and more potent in reducing the expression of TNF-alpha mRNA, as well as reducing serum TNF-alpha levels. Furthermore, the ASO/GLC complex successfully rescued animals from fulminant hepatitis and mortality. Compared to naked ASO, the complex notably reduced the dose administrated in animals and prolonged its effectiveness. A single dose of 5 mg ASO per kg body weight achieved a satisfactory effect after 5 days, and 20 mg ASO per kg body weight preserved 70% of the effect after more than 2 weeks. Its efficacy was affirmed through both pretreatment and therapeutic use after liver damage had begun. CONCLUSIONS: Inhibiting TNF-alpha expression in the liver by this strategy represents a novel therapeutic approach that may be valuable for the treatment of some inflammation-related liver diseases. PMID- 19189286 TI - Physiological origin of low-frequency drift in blood oxygen level dependent (BOLD) functional magnetic resonance imaging (fMRI). AB - We investigated the biophysical mechanism of low-frequency drift in blood-oxygen level-dependent (BOLD) functional magnetic resonance imaging (fMRI) (0.00-0.01 Hz), by exploring its spatial distribution, dependence on imaging parameters, and relationship with task-induced brain activation. Cardiac and respiratory signals were concurrently recorded during MRI scanning and subsequently removed from MRI data. It was found that the spatial distribution of low-frequency drifts in human brain followed a tissue-specific pattern, with greater drift magnitude in the gray matter than in white matter. In gray matter, the dependence of drift magnitudes on TE was similar to that of task-induced BOLD signal changes, i.e., the absolute drift magnitude reached the maximum when TE approached T(2)* whereas relative drift magnitude increased linearly with TE. By systematically varying the flip angle, it was found that drift magnitudes possessed a positive dependence on image intensity. In phantom experiments, the observed drift was not only much smaller than that of human brain, but also showed different dependence on TE and flip angle. In fMRI studies with visual stimulation, a strong positive correlation between drift effects at baseline and task-induced BOLD signal changes was observed both across subjects and across activated pixels within individual participants. We further demonstrated that intrinsic, physiological drift effects are a major component of the spontaneous fluctuations of BOLD fMRI signal within the frequency range of 0.0-0.1 Hz. Our study supports brain physiology, as opposed to scanner instabilities or cardiac/respiratory pulsations, as the main source of low-frequency drifts in BOLD fMRI. PMID- 19189287 TI - Ultra-low output impedance RF power amplifier for parallel excitation. AB - Inductive coupling between coil elements of a transmit array is one of the key challenges faced by parallel RF transmission. An ultra-low output impedance RF power amplifier (PA) concept was introduced to address this challenge. In an example implementation, an output-matching network was designed to transform the drain-source impedance of the metallic oxide semiconductor field effect transistor (MOSFET) into a very low value for suppressing interelement coupling effect, and meanwhile, to match the input impedance of the coil to the optimum load of the MOSFET for maximizing the available output power. Two prototype amplifiers with 500-W output rating were developed accordingly, and were further evaluated with a transmit array in phantom experiments. Compared to the conventional 50-Omega sources, the new approach exhibited considerable effectiveness suppressing the effects of interelement coupling. The experiments further indicated that the isolation performance was comparable to that achieved by optimized overlap decoupling. The new approach, benefiting from a distinctive current-source characteristic, also exhibited a superior robustness against load variation. Feasibility of the new approach in high-field MR was demonstrated on a 3T clinical scanner. PMID- 19189288 TI - Balanced multipoint displacement encoding for DENSE MRI. AB - Displacement encoding with stimulated echoes (DENSE) is a quantitative imaging technique that encodes tissue displacement in the phase of the acquired signal. Various DENSE sequences have encoded displacement using methods analogous to the simple multipoint methods of phase contrast (PC) MRI. We developed general n dimension balanced multipoint encoding for DENSE. Using these methods, phase noise variance decreased experimentally by 73.7%, 65.6%, and 61.9% compared with simple methods, which closely matched the theoretical decreases of 75%, 66.7%, and 62.5% for one-dimensional (1D), 2D, and 3D encoding, respectively. Phase noise covariances decreased by 99.2% and 99.3% for balanced 2D and 3D encoding, consistent with the zero-covariance prediction. The direction bias inherent to the simple methods was decreased to almost zero using balanced methods. Reduced phase noise and improved displacement and strain maps using balanced methods were visually observed in phantom and volunteer images. Balanced multipoint encoding can also be applied to PC MRI. PMID- 19189289 TI - MRI contrast using solid-state, B1-distorting, microelectromechanical systems (MEMS) microresonant devices (MRDs). AB - Presently, signal generation in MRI depends on the concentration and relaxivity of protons or other MR-active nuclei, and contrast depends on local differences in signal. In this proof-of-principle study, we explore the use of nonchemical, solid-state devices for generating detectable signal and/or contrast in vitro and in vivo. We introduce the concept of microresonant devices (MRDs), which are micron-sized resonators fabricated using microelectromechanical systems (MEMS) technology. Fifteen-micrometer (15-microm)-thick, coil MRDs were designed to resonate at the 3T Larmor frequency of protons (127.7 MHz) and were fabricated using tantalum (Ta) oxide thin-film capacitors and copper-plated spiral inductors. The performance of MRDs having final diameters of 300, 500, and 1000 microm were characterized in saline using a radio frequency (RF) scanning microscope and a clinical 3T MR scanner. The measured B(1) fields of 300 microm to 1000 microm MRDs ranged from 3.25 microT to 3.98 microT, and their quality factors (Q) ranged from 3.9 to 7.2. When implanted subcutaneously in the flank of a mouse, only MRDs tuned to the resonant frequency of protons generated a measurable in vivo B(1) field. This study lays the foundation for a new class of solid-state contrast agents for MRI. PMID- 19189290 TI - Articular cartilage deformation determined in an intact tibiofemoral joint by displacement-encoded imaging. AB - This study demonstrates the in vitro displacement and strain of articular cartilage in a cyclically-compressed and intact joint using displacement-encoded imaging with stimulated echoes (DENSE) and fast spin echo (FSE). Deformation and strain fields exhibited complex and heterogeneous patterns. The displacements in the loading direction ranged from -1688 to -1481 microm in the tibial cartilage and from -1601 to -764 microm in the femoral cartilage. Corresponding strains ranged from -9.8% to 0.7% and from -4.3% to 0.0%. The displacement and strain precision were determined to be 65 microm and less than 0.2%, respectively. Displacement-encoded magnetic resonance imaging is capable of determining the nonuniform displacements and strains in the articular cartilage of an intact joint to a high precision. Knowledge of these nonuniform strains is critical for the in situ characterization of normal and diseased tissue, as well as the comprehensive evaluation of repair constructs designed using regenerative medicine. PMID- 19189291 TI - Novel reconstruction method for three-dimensional axial continuously moving table whole-body magnetic resonance imaging featuring autocalibrated parallel imaging GRAPPA. AB - Continuously moving table MR imaging has been successfully evaluated for whole body tumor staging and metastasis screening. In previous studies it was demonstrated that three-dimensional (3D) slab-selective excitation with lateral readout can provide very efficient k-space coverage when the longitudinal field of view (FOV) is limited. To reduce respiratory artifacts, data acquisition in the thoracoabdominal region of the patient typically must be performed during one single breath hold. This consequently restricts acquisition time and thus spatial resolution. In this work, a novel reconstruction method is introduced for axial 3D moving table data acquisition with lateral readout. The method features table position correction completely in k-space and is compatible with autocalibrated parallel imaging (GRAPPA). Parallel imaging can be applied to increase spatial resolution while maintaining the breath-holding time. A sophisticated protocol for whole-body moving table MRI was developed. The impact of gradient nonlinearity on the featured imaging method was evaluated in phantom and volunteer experiments. Finally, the protocol was optimized toward minimizing residual artifacts. Moving table whole-body MRI with lateral readout was performed in 5 healthy volunteers and was compared with lateral readout data acquired with a GRAPPA accelerated protocol providing increased spatial resolution. PMID- 19189292 TI - Delta relaxation enhanced MR: improving activation-specificity of molecular probes through R1 dispersion imaging. AB - MR molecular imaging enables high-resolution, in vivo study of molecular processes frequently utilizing gadolinium-based probes that specifically bind to a particular biological molecule or tissue. While some MR probes are inactive when unbound and produce enhancement only after binding, the majority are less specific and cause enhancement in either state. Accumulation processes are then required to increase probe concentration in regions of the target molecule/tissue. Herein, a method is described for creating specificity for traditionally nonspecific probes. This method utilizes MR field-cycling methods to produce MRI contrast related to the dependence of R(1) upon magnetic field. It is shown that the partial derivative of R(1) with respect to magnetic field strength, R(1)', can be used as an unambiguous measure of probe binding. T(1) weighted images and R(1)' images were produced for samples of albumin and buffer both enhanced with the albumin-binding agent Vasovist. For T(1) images, samples with low concentrations of Vasovist in an albumin solution could not be differentiated from samples with higher concentrations of Vasovist in buffer. Conversely, the R(1)' images showed high specificity to albumin. Albumin samples with a 10-microM concentration of Vasovist were enhanced over buffer samples containing up to 16 times more Vasovist. PMID- 19189293 TI - Fast acquisition scheme for achieving high-resolution MRS with J-scaling under inhomogeneous fields. AB - Intermolecular multiple-quantum coherences (iMQCs) can refocus the phase dispersion caused by magnetic field inhomogeneities while preserving the chemical shift, so they have been applied to achieve high-resolution MR spectroscopy free of line broadening caused by susceptibility gradients. However, previous iMQC high-resolution methods all require two-dimensional spectra sampling of the full range of chemical shifts of solute evolutions in both F(1) and F(2) dimensions, resulting in a prolonged scanning time for data acquisition. In this work, sparse sampling in the t(1) dimension and subsequent fold-over correction are used to speed up the intermolecular zero-quantum coherence spectroscopy by up to 50 times on high-field MR systems. Furthermore, three types of spectra with homo decoupling, original J-coupling constants, and doubled J-coupling constants respectively are obtained with manipulation of the t(1) period. The water suppression is also improved by the combined use of intermolecular double-quantum filter and excitation sculpting. The feasibilities of this group of new sequences are demonstrated by experiments using an agar gel phantom with an air bubble, in vitro pig brain tissues and an intact postmortem mudskipper. PMID- 19189294 TI - Postmortem MRI of human brain hemispheres: T2 relaxation times during formaldehyde fixation. AB - Unlike in vivo imaging, postmortem MRI allows for invasive examination of the tissue specimen immediately after the MR scan. However, natural tissue decomposition and chemical fixation cause the postmortem tissue's MRI properties to be different from those found in vivo. Moreover, these properties change as postmortem fixation time elapses. The goal of this study was to characterize the T(2) relaxation changes that occur over time in cadaveric human brain hemispheres during fixation. Five hemispheres immersed in formaldehyde solution were scanned on a weekly basis for 3 months postmortem, and once again at 6 months postmortem. The T(2) relaxation times were measured throughout the hemispheres. Over time, T(2) values near the edges of the hemispheres decreased rapidly after death, while T(2) values of deep tissue decreased more slowly. This difference is likely due to the relatively large distance from the hemisphere surface, and other barriers limiting diffusion of formaldehyde molecules to deep tissues. In addition, T(2) values in deep tissue did not continuously decay to a plateau, but instead reached a minimum and then increased to a plateau. This final increase may be due to the effects of prolonged tissue decomposition, a hypothesis that is supported by numerical simulations of the fixation process. PMID- 19189295 TI - Implementation of quantitative perfusion imaging using pulsed arterial spin labeling at ultra-high field. AB - This study compares the implementation of the STAR and FAIR pulsed arterial spin labeling (PASL) schemes to form quantitative perfusion maps at ultra-high field, 7 Tesla (T), and high field, 3 T. Phantom experiments were performed to compare the inversion efficiency and profile of the labeling pulses at 7 T and 3 T and to optimize in-plane saturation techniques. The perfusion weighted (PW) signal was measured at a range of postlabeling delay times and quantitative perfusion maps were calculated on a voxel-by-voxel basis. An increase in PW signal was found with field strength, and together with the increased signal-to-noise ratio, this led to improved image signal-to-noise and quality of fit of perfusion maps at 7 T. PMID- 19189296 TI - Feasibility of concurrent dual contrast enhancement using CEST contrast agents and superparamagnetic iron oxide particles. AB - A major challenge for cellular and molecular MRI is to study interactions between two different cell populations or biological processes. We studied the possibility to simultaneously image contrast agents based on two different MRI contrast mechanisms: chemical exchange saturation transfer (CEST) and enhancement of T2 relaxation. Various amounts of superparamagnetic iron oxide (SPIO) nanoparticles were mixed with a fixed concentration (250 microM) of the CEST agent poly-L-lysine. T2 maps, CEST maps, and frequency-dependent saturation spectra were then measured. Color-coded overlay maps demonstrated the feasibility of concurrent dual contrast enhancement. We found that at concentrations lower than 5 microg(Fe)/mL both contrast agents can be imaged simultaneously. At higher concentrations, the iron-based agent can be used to "shut off" the signal arising from the CEST agent. These initial findings are a first step toward using dual CEST/T2 contrast imaging for studying multiple cellular or molecular targets simultaneously in vivo. PMID- 19189297 TI - Oleanane triterpenoid CDDO-Me inhibits growth and induces apoptosis in prostate cancer cells by independently targeting pro-survival Akt and mTOR. AB - BACKGROUND: Synthetic triterpenoids are potent anticancer agents, but their therapeutic efficacy or mechanism of action for prostate cancer has not been investigated. The goal of this study was to determine the antitumor activity and the mechanism of action of methyl-2-cyano-3,12-dioxooleana-1,9(11)-dien-28-oate (CDDO-Me), a oleanane-derived synthetic triterpenoid for human prostate cancer cells. METHODS: The antitumor activity of CDDO-Me for hormone-refractory PC-3 (AR(-)) and C4-2 (AR(+)) prostate cancer cell lines was determined by effects on cell growth and induction of apoptosis, identification of molecular targets, and therapeutic efficacy in vivo in PC-3 xenograft model. RESULTS: CDDO-Me inhibited the growth and induced apoptosis in PC-3 and C4-2 cells at extremely low concentrations. The antitumor activity of CDDO-Me was associated with the inhibition of p-Akt, mammalian target of rapamycin (mTOR), and nuclear factor kappa B (NF-kappaB) signaling proteins and their downstream targets such as p-Bad and p-Foxo3a (Akt); p-S6K1, p-eIF-4E and p-4E-BP1 (mTOR); and COX-2, VEGF and cyclin D1(NF-kappaB). Silencing of Akt sensitized the PC-3 cells to CDDO-Me, whereas overexpression of Akt induced resistance to CDDO-Me. Targeted silencing of Akt showed that Akt does not regulate mTOR activation in PC-3 cells, but targeted silencing of mTOR sensitized PC-3 cells to CDDO-Me mediated growth inhibition. Further, treatment with CDDO-Me inhibited the growth of PC-3 xenografts in nude mice. CONCLUSIONS: This study demonstrated potent antitumor activity of CDDO-Me against prostate cancer cells both in vitro and in vivo. Data also identified Akt and mTOR as molecular targets of CDDO-Me in prostate cancer cells. PMID- 19189298 TI - Inflammatory cutaneous adverse effects of methylene blue dye injection for lymphatic mapping/sentinel lymphadenectomy. AB - BACKGROUND: Methylene blue (MB) dye has been used for lymphatic mapping/sentinel lymphadenectomy (LM/SL) in staging of melanoma and breast cancer. It has been noted to cause skin necrosis, but its more mild adverse effects from intraparenchymal breast injections are not well characterized. METHODS: Patients undergoing LM/SL for breast cancer and melanoma were reviewed, with attention devoted to skin manifestations. Patients undergoing mastectomies were excluded to rule out changes from flap devascularization. All breast patients were injected intraparenchymally. RESULTS: Ninety-five patients underwent MB injection during a nationwide shortage of lymphazurin; 78 for breast cancer and 17 for melanoma, with 51 patients undergoing breast conservation (BCT). There was no frank skin necrosis among any of the patients. Six (11.8%) BCT patients demonstrated inflammatory changes. Four patients developed findings indistinguishable from infectious cellulitis, with two developing skin telangiectasias prior to radiotherapy. Two patients had fat necrosis confirmed at the MB injection site away from the surgical site; one on imaging and one by biopsy. Most symptoms resolved after conservative management. CONCLUSIONS: MB dye may cause cutaneous changes more subtle than previously described. Physicians caring for patients having LM/SL using MB should be aware of these effects so that a proper differential diagnosis can be entertained postoperatively. PMID- 19189299 TI - Significance of preoperative HbA1c level in patients with diabetes mellitus and clinically localized prostate cancer. AB - INTRODUCTION: We investigated potential relationships of history of diabetes mellitus (DM) and glycemic control, represented by hemoglobin A1c (HbA1c) level, with characteristics of tumor among patients who received radical prostatectomy (RP) for clinically localized prostate cancer. METHODS: We reviewed data of 740 patients who underwent RP for clinically localized prostate cancer between 2004 and 2008 without receiving preoperative radiation or hormonal treatment. Univariate and multivariate logistic regression analyses addressed the associations of history of DM and HbA1c level with known prognostic variables of prostate cancer. RESULTS: No significant differences were observed in various preoperative and pathological parameters between those with (n = 89) and without DM (n = 651). When only the subjects with DM were divided into two groups (group 1 and 2) according to HbA1c level (<6.5% vs. >/=6.5%), group 2 demonstrated significantly higher rate of extraprostatic extension of tumor (P = 0.043) and high (>/=7) pathological Gleason score (P = 0.005) than group 1. Also among those with DM, HbA1c level was observed to be an independent predictor for high pathologic Gleason score (P = 0.010) and extraprostatic extension of tumor (P = 0.035), respectively in multivariate analyses. CONCLUSION: Although simple history of having DM may not be a significant factor regarding aggressiveness of clinically localized prostate cancer, the glycemic control, as represented by HbA1c level, may be a useful preoperative predictor of aggressive tumor profile among patients with DM who are also diagnosed with clinically localized prostate cancer. PMID- 19189300 TI - Morphofunctional study of 12-O-tetradecanoyl-13-phorbol acetate (TPA)-induced differentiation of U937 cells under exposure to a 6 mT static magnetic field. AB - This study deals with the morphofunctional influence of 72 h exposure to a 6 mT static magnetic field (SMF) during differentiation induced by 50 ng/ml 12-O tetradecanoyl-13-phorbol acetate (TPA) in human leukaemia U937 cells. The cell morphology of U937 cells was investigated by optic and electron microscopy. Specific antibodies and/or molecules were used to label CD11c, CD14, phosphatidylserine, F-actin and to investigate the distribution and activity of lysosomes, mitochondria and SER. [Ca(2+)](i) was evaluated with a spectrophotometer. The degree of differentiation in SMF-exposed cells was lower than that of non-exposed cells, the difference being exposure time-dependent. SMF exposed cells showed cell shape and F-actin modification, inhibition of cell attachment, appearance of membrane roughness and large blebs and impaired expression of specific macrophagic markers on the cell surface. The intracellular localization of SER and lysosomes was only partially affected by exposure. A significant localization of mitochondria with an intact membrane potential at the cell periphery in non-exposed, TPA-stimulated cells was observed; conversely, in the presence of SMF, mitochondria were mainly localised near the nucleus. In no case did SMF exposure affect cell viability. The sharp intracellular increase of [Ca(2+)](i) could be one of the causes of the above-described changes. PMID- 19189301 TI - Quantitative RT-PCR analysis of estrogen receptor gene expression in laser microdissected prostate cancer tissue. AB - BACKGROUND: Real-time quantitative RT-PCR analysis of laser microdissected tissue is considered the most accurate technique for determining tissue gene expression. The discovery of estrogen receptor beta (ERbeta) has focussed renewed interest on the role of estrogen receptors in prostate cancer, yet few studies have utilized the technique to analyze estrogen receptor gene expression in prostate cancer. METHODS: Fresh tissue was obtained from 11 radical prostatectomy specimens and from 6 patients with benign prostate hyperplasia. Pure populations of benign and malignant prostate epithelium were laser microdissected, followed by RNA isolation and electrophoresis. Quantitative RT-PCR was performed using primers for androgen receptor (AR), estrogen receptor beta (ERbeta), estrogen receptor alpha (ERalpha), progesterone receptor (PGR) and prostate specific antigen (PSA), with normalization to two housekeeping genes. Differences in gene expression were analyzed using the Mann-Whitney U-test. Correlation coefficients were analyzed using Spearman's test. RESULTS: Significant positive correlations were seen when AR and AR-dependent PSA, and ERalpha and ERalpha-dependent PGR were compared, indicating a representative population of RNA transcripts. ERbeta gene expression was significantly over-expressed in the cancer group compared with benign controls (P < 0.01). In contrast, PGR expression was significantly down-regulated in the cancer group (P < 0.05). There were no significant differences in AR, ERalpha or PSA expression between the groups. This study represents the first to show an upregulation of ERbeta gene expression in laser microdissected prostate cancer specimens. CONCLUSIONS: In concert with recent studies the findings suggest differential production of ERbeta splice variants, which may play important roles in the genesis of prostate cancer. PMID- 19189303 TI - Prostatic chondroitin sulfate is increased in patients with metastatic disease but does not predict survival outcome. AB - BACKGROUND: Previous studies from our laboratory demonstrated a strong association between an elevated level of chondroitin sulfate (CS) in peritumoral stroma and PSA-relapse in patients with early stage disease. In this study we determined whether CS levels could predict overall survival in men diagnosed with advanced prostate cancer subsequently treated by orchiectomy alone. METHODS: CS was localized in archived prostatic tissues by immunohistochemistry, and the level of CS expression as measured by video image analysis was compared in cohorts of 157 and 60 men with early stage or advanced disease, respectively. RESULTS: The CS levels in the peritumoral stroma of patients without relapse after treatment for early stage disease was significantly reduced compared to levels in prostate tissue from patients who either relapsed (P = 0.003) or were diagnosed with advanced prostate cancer (P < 0.00001). There was no difference between the median CS level in the peritumoral prostatic stroma of early stage patients that relapsed after treatment and patients diagnosed with advanced prostate cancer. Increased CS levels (P < 0.0001) and high Gleason score (P < 0.0001) were associated with an increased rate of PSA-relapse in the cohort of patients with early stage disease. However, neither CS level nor Gleason score alone or in combination could predict survival outcome in patients with advanced prostate cancer following androgen deprivation therapy. CONCLUSIONS: Although peritumoral CS levels and Gleason score are strong predictors of relapse-free survival in early stage prostate cancer patients, neither peritumoral CS levels nor Gleason score can predict survival outcome in patients with advanced disease. PMID- 19189302 TI - Countertransference reactions to adolescents with eating disorders: relationships to clinician and patient factors. AB - OBJECTIVE: Clinical report suggests that therapists have strong and sometimes difficult-to-manage reactions to patients with eating disorders (EDs); however, systematic research is largely absent. The purpose of this study was to explore the emotional responses, or countertransference (CT) reactions, clinicians experience when working with patients with EDs, and to identify clinician, patient, and therapy variables associated with these responses. METHOD: One hundred twenty clinicians reported on multiple variables related to an adolescent female patient they were treating for an ED. RESULTS: Six patterns of reactions were identified: angry/frustrated, warm/competent, aggressive/sexual, failing/incompetent, bored/angry at parents and overinvested/worried feelings. The factors showed meaningful relationships across clinician demographics, patient characteristics, and treatment techniques. DISCUSSION: Overall, clinician's reactions were most frequently associated with the clinician's gender, patient's level of functioning and improvement during treatment, and patient personality style. These issues have important implications for treatment, training and supervision. PMID- 19189304 TI - Vasoactive intestinal peptide behaves as a pro-metastatic factor in human prostate cancer cells. AB - BACKGROUND: There is little known on the involvement of vasoactive intestinal peptide (VIP) in the metastatic cascade of human prostate cancer, that is, cell proliferation, cell-cell adhesion, extracellular-matrix degradation, and migration/invasion. Here we evaluated the expression of related biomarker proteins (cyclin D1, metalloproteinases MMP-2 and MMP-9, and E-cadherin) in human androgen-dependent (LNCaP) and independent (PC3) prostate cancer cells. METHODS: Reverse transcriptase (RT)-polymerase chain reaction (PCR), gelatin zymography, Western blotting, confocal immunofluorescence microscopy, and assays on cell proliferation, adhesion, wound-healing, migration and random homing were performed. RESULTS: VIP increased cell proliferation and cyclin D1 expression whereas it decreased cell adhesion and E-cadherin expression in LNCaP and PC3 cells. VIP enhanced the gelatinolytic activity of MMP-2 and MMP-9. Semiquantitative RT-PCR assays showed that VIP stimulated mRNA levels of these MMPs and suppressed mRNA levels of its inhibitory protein RECK. VIP promoted cell invasion and migration, and the responses were faster according to the most aggressive status in cancer progression (androgen-independence). The involvement of nuclear factor-kappaB (NF-kappaB) was demonstrated since the anti-inflammatory agent curcumin blocked VIP effects on the above biomarkers in both cell lines. CONCLUSIONS: Taken together, these results and the presence of kappaB sites on gene promoter of cyclin D1, MMPs and, possibly, E-cadherin suggest that VIP may act as a cytokine in an early metastatic stage of human prostate cancer through the NF-kappaB/MMPs-RECK/E-cadherin system. Our findings may help to define novel targets and agents with potential usefulness in prostate cancer therapy. PMID- 19189305 TI - Low stroma androgen receptor level in normal and tumor prostate tissue is related to poor outcome in prostate cancer patients. AB - BACKGROUND: The role of androgen receptors (ARs) in the prostate tumor cell environment is largely unknown. METHODS: AR immunostaining was evaluated in relation to stroma morphology, expression of AR co-activator ARA55, tumor characteristics and clinical outcome in normal and prostate cancer (PCa) tissue obtained at transurethral resection in men treated with expectancy, and in diagnostic transrectal core biopsies in men treated with surgical castration. Stroma composition was studied by Masson-trichrome and desmin staining. Levels of AR and ARA55 mRNA were quantified by laser micro-dissection and RT-PCR. RESULTS: The percentage of cells with positive nuclear AR immunostaining in the tumor and normal stroma was inversely related to Gleason score, tumor size, tumor stage, metastasis, response to castration therapy, and cancer-specific survival. The AR staining in the normal stroma provided independent prognostic information in Cox multiple linear regression analysis. Loss of stroma AR staining was linked to low expression of ARA55 in stroma smooth muscle cells, and in tumors also to gradual disappearance of this cell type. CONCLUSIONS: PCa aggressiveness and efficacy of castration therapy are related to AR levels in the tumor stroma and importantly to AR levels in the surrounding normal prostate tissue stroma. . PMID- 19189306 TI - Selective expression of CD44, a putative prostate cancer stem cell marker, in neuroendocrine tumor cells of human prostate cancer. AB - BACKGROUND: Hormonal therapy is effective for advanced prostate cancer (PC) but the disease often recurs and becomes hormone-refractory. It is hypothesized that a subpopulation of cancer cells, that is, cancer stem cells (CSCs), survives hormonal therapy and leads to tumor recurrence. CD44 expression was shown to identify tumor cells with CSC features. PC contains secretory type epithelial cells and a minor population of neuroendocrine cells. Neuroendocrine cells do not express androgen receptor and are quiescent, features associated with CSCs. The purpose of the study was to determine the expression of CD44 in human PC and its relationship to neuroendocrine tumor cells. METHODS: Immunohistochemistry and immunofluorescence were performed to study CD44 expression in PC cell lines, single cells from fresh PC tissue and archival tissue sections of PC. We then determined if CD44+ cells represent neuroendocrine tumor cells. RESULTS: In human PC cell lines, expression of CD44 is associated with cells of NE phenotype. In human PC tissues, NE tumor cells are virtually all positive for CD44 and CD44+ cells, excluding lymphocytes, are all NE tumor cells. CONCLUSIONS: Selective expression of the stem cell-associated marker CD44 in NE tumor cells of PC, in combination with their other known features, further supports the significance of such cells in therapy resistance and tumor recurrence. PMID- 19189307 TI - Changes in the GST activity of the mussel Mytilus galloprovincialis during exposure and depuration of microcystins. AB - Mussels are quite resistant to cyanotoxins and their resistance may be because of an efficient metabolization of cyanotoxins by glutathione-S-transferases (GST) activity. Nevertheless, other secondary metabolites may interfere with the detoxication efficiency. The accumulation and depuration of hepatotoxins produced by the freshwater cyanobacterium Microcystis aeruginosa in the mussel Mytilus galloprovincialis were studied. Mussels were fed twice a day 1.5x10(5) cells/mL of the toxic cyanobacterium, which produces microcystins (MCs) -FR, -LR and -WR, for 4 days. After that period, the animals were placed in toxin-free water and were fed the green algae Ankistrodesmus sp. During 2 weeks, the concentration of the toxin in the mussels was monitored using an ELISA assay. Mussels showed a maximum detectable level of MCs of 0.38 microg/g mussels dry weight (DW) during the accumulation period and 0.37 microg MC/g mussel DW by day 4 of the depuration period. Then there was a decrease trend with peaks of toxin at days 8 and 12 of the depuration period. The activity of the detoxication enzymes GST was studied and the results showed that the peaks of toxin in the mussels coincide with an increase in the activity of GST. These results support the hypothesis that the rise of the toxin level on days 4, 8 and 12 of the depuration period in the mussels may be related to the renewal of protein phosphatases and subsequent release of unbound toxins. PMID- 19189308 TI - Evaluation of antral follicle growth in the macaque ovary during the menstrual cycle and controlled ovarian stimulation by high-resolution ultrasonography. AB - To date, ultrasonography of monkey ovaries is rare and typically of low resolution. The objectives of this study were to use state-of-the-art, high resolution, transabdominal ultrasonography with real-time Doppler capabilities to: (1) determine whether one can reliably detect in real time the large dominant follicle, the corpus luteum (CL), and small (<2 mm) antral follicles on the ovaries of rhesus monkeys during the natural menstrual cycle; and (2) predict the follicular response of rhesus ovaries to controlled ovarian stimulation (COS) protocols. Rhesus monkeys were selected for transabdominal ultrasonography using a GE Voluson 730 Expert Doppler System at discrete stages of the menstrual cycle. Subsequently, serial ultrasound scanning was employed to observe growth of antral follicles and the CL. Finally, females were scanned to assess follicular growth during COS. The dominant structure and small antral follicles (<2 mm) were reliably visualized in real time. The follicle destined to ovulate could be identified by size differential by day 3 of the follicular phase. The number of small antral follicles present before onset of COS protocol correlated positively with the number of metaphase II-stage oocytes collected after treatment. The results of this study demonstrate that the population dynamics of antral follicle pools can be noninvasively evaluated in monkeys during natural and pharmacologic ovarian cycles. PMID- 19189309 TI - The CDK domain of p21 is a suppressor of IL-1beta-mediated inflammation in activated macrophages. AB - Significant morbidity and mortality can be attributed to inflammatory diseases; therefore, a greater understanding of the mechanisms involved in the progression of inflammation is crucial. Here, we demonstrate that p21((WAF1/CIP1)), an established suppressor of cell cycle progression, is a inhibitor of IL-1beta synthesis in macrophages. Mice deficient in p21 (p21(-/-)) display increased susceptibility to endotoxic shock, which is associated with increased serum levels of IL-1beta. Administration of IL-1 receptor antagonist reduces LPS induced lethality in p21(-/-) mice. Analysis of isolated macrophages, which are one of the central producers of IL-1beta, reveals that deficiency for p21 led to more IL-1beta mRNA and pro-protein synthesis following TLR ligation. The increase in IL-1beta pro-protein is associated with elevated secretion of active IL-1beta by p21(-/-) macrophages. siRNA-mediated knockdown of p21 in human macrophages results in increased IL-1beta secretion as well. A peptide mapping strategy shows that the cyclin-dependent-kinase (CDK)-binding domain of p21 is sufficient to reduce the secretion of IL-1beta by p21(-/-) macrophages. These data suggest a novel role for p21 and specifically for the CDK-binding domain of p21((WAF1/CIP1)) in inhibiting inflammation. PMID- 19189310 TI - DMBT1 functions as pattern-recognition molecule for poly-sulfated and poly phosphorylated ligands. AB - Deleted in malignant brain tumors 1 (DMBT1) is a secreted glycoprotein displaying a broad bacterial-binding spectrum. Recent functional and genetic studies linked DMBT1 to the suppression of LPS-induced TLR4-mediated NF-kappaB activation and to the pathogenesis of Crohn's disease. Here, we aimed at unraveling the molecular basis of its function in mucosal protection and of its broad pathogen-binding specificity. We report that DMBT1 directly interacts with dextran sulfate sodium (DSS) and carrageenan, a structurally similar sulfated polysaccharide, which is used as a texturizer and thickener in human dietary products. However, binding of DMBT1 does not reduce the cytotoxic effects of these agents to intestinal epithelial cells in vitro. DSS and carrageenan compete for DMBT1-mediated bacterial aggregation via interaction with its bacterial-recognition motif. Competition and ELISA studies identify poly-sulfated and poly-phosphorylated structures as ligands for this recognition motif, such as heparansulfate, LPS, and lipoteichoic acid. Dose-response studies in Dmbt1(-/-) and Dmbt1(+/+) mice utilizing the DSS-induced colitis model demonstrate a differential response only to low but not to high DSS doses. We propose that DMBT1 functions as pattern recognition molecule for poly-sulfated and poly-phosphorylated ligands providing a molecular basis for its broad bacterial-binding specificity and its inhibitory effects on LPS-induced TLR4-mediated NF-kappaB activation. PMID- 19189311 TI - CTL induction by cross-priming is restricted to immunodominant epitopes. AB - CTL are induced by two pathways, i.e. direct priming, where tumor cells present tumor antigens to naive specific CTL, and cross-priming, where professional APC cross-present captured tumor antigens to CTL. Here, we examined direct priming versus cross-priming after immunizing (H-2(b) x H-2(d)) F1 mice with either H 2(b) or H-2(d) positive tumor cells transfected with the GP or nucleoprotein (NP) of lymphocytic choriomeningitis virus (LCMV). Cross-priming was observed for the immunodominant epitopes LCMV-gp33 and -np118, although direct induction resulted in higher CTL frequencies. In contrast, CTL specific for the subdominant epitopes LCMV-gp283 or -np396 were induced only if epitopes were presented directly on MHC class I molecules of the immunizing cell. The broader repertoire and the higher CTL frequencies induced after vaccination with haplotype-matched tumor cells resulted in more efficient anti-tumor and antiviral protection. Firstly, our results indicate that certain virus and tumor antigens may not be detected by CD8(+) T cells because of impaired cross-priming. Secondly, efficient cross priming contributes to the immunodominant nature of a tumor-specific CTL epitope. Thirdly, vaccine strategies using autologous or syngenic antigen-expressing cells induce a broader repertoire of tumor-specific CTL and higher CTL frequencies. PMID- 19189312 TI - Advances in transmission electron microscopy: in situ straining and in situ compression experiments on metallic glasses. AB - In the field of transmission electron microscopy (TEM), fundamental and practical reasons still remain that hamper a straightforward correlation between microscopic structural information and deformation mechanisms in materials. In this article, it is argued that one should focus in particular on in situ rather than on postmortem observations of the microstructure. This viewpoint has been exemplified with in situ straining and in situ compression studies on metallic glasses. In situ TEM straining of amorphous metals permits an evaluation of the thickness of the liquid-like layer (LLL) formed because of heat evolution after shear band development. The experimental evaluation confirms that the thickness of a LLL present at the last moment of fracture substantially exceeds the generally accepted thickness of a shear band. In situ TEM and in situ SEM compression experiments on metallic glass pillars lead to the conclusion that smaller sized pillars deform more homogeneously than larger sized pillars. PMID- 19189313 TI - In situ deformation of thin films on substrates. AB - Metallic thin-film plasticity has been widely studied by using the difference between the coefficients of thermal expansion of the film and the underlying substrate to induce stress. This approach is commonly known as the wafer curvature technique, based on the Stoney equation, which has shown that thinner films have higher yield stresses. The linear increase of the film strength as a function of the reciprocal film thickness, down to a couple hundred nanometers, has been rationalized in terms of threading and interfacial dislocations. Polycrystalline films also show this kind of dependence when the grain size is larger than or comparable to the film thickness. In situ TEM performed on plan view or cross-section specimens faithfully reproduces the stress state and the small strain levels seen by the metallic film during wafer curvature experiments and simultaneously follows the change in its microstructure. Although plan-view experiments are restricted to thinner films, cross-sectional samples where the film is reduced to a strip (or nanowire) on its substrate are a more versatile configuration. In situ thermal cycling experiments revealed that the dislocation/interface interaction could be either attractive or repulsive depending on the interfacial structure. Incoherent interfaces clearly act as dislocation sinks, resulting in a dislocation density drop during thermal cycles. In dislocation-depleted films (initially thin or annealed), grain boundaries can compensate for the absence of dislocations by either shearing the film similarly to threading dislocations or through fast diffusion processes. Conversely, dislocations are confined inside the film by image forces in the cases of epitaxial interfaces on hard substrates. To increase the amount of strain seen by a film, and to decouple the effects of stress and temperature, compliant substrates can also be used as support for the metallic film. The composite can be stretched at a given temperature using heating/cooling straining holders. Other in situ TEM methods that served to reveal scaling effects are also reviewed. Finally, an alternate method, based on a novel bending holder that can stretch metallic films on rigid substrates, is presented. PMID- 19189314 TI - Combining in situ transmission electron microscopy and molecular dynamics computer simulations to reveal the interaction mechanisms of dislocations with stacking-fault tetrahedron in nuclear materials. AB - In situ straining experiments conducted in a transmission electron microscope have been performed to reveal how dislocations interact with and annihilate isolated stacking-fault tetrahedra, common defect clusters in irradiated face centered cubic metals. Comparison of the experimental results with the predictions from molecular dynamics computer simulations shows some similarities and some striking differences. The comparison suggests challenges for the simulations and demonstrates how, despite the disparity in operational parameters, the simulations can be used to interpret experimental results. PMID- 19189315 TI - Adenoviral delivery of dominant-negative transforming growth factor beta type II receptor up-regulates transcriptional repressor SKI-like oncogene, decreases matrix metalloproteinase 2 in hepatic stellate cell and prevents liver fibrosis in rats. AB - BACKGROUND: Dominant-negative transforming growth factor beta type II receptor (TbetaRIIDeltacyt) is a protein that blocks transforming growth factor (TGF-beta) signaling. Because the consequences of blocking TGF-beta have not been completely elucidated in liver fibrosis, we analysed the effects of adenoviral delivery of TbetaRIIDeltacyt on profibrogenic genes and matrix metalloproteinase (MMP) proteins, as well as on TGF-beta signal repressor SKI-like oncogene (SnoN), in cultured hepatic stellate cells (HSCs) and in a rat model of liver fibrosis. METHODS: To induce liver fibrosis, rats were treated with thioacetamide for 7 weeks and administrated once with Ad-TbetaRIIDeltacyt or Ad-betagal through the iliac vein. Fibrosis was measured by morphometric analysis. We evaluated SnoN by western blot, immunocytochemistry and immunohistochemistry; MMP activity was determined by zymography and profibrogenic gene expression by the real-time reverse transcriptase-polymerase chain reaction in cultured HSCs and liver tissue. RESULTS: Profibrogenic gene expression of collagen alpha1 (I), TGF-beta1, platelet-derived growth factor-B, plasminogen activator inhibitor (PAI)-1, tissue inhibitor of matrix metalloproteinase-1 and MMP-2 was down-regulated; whereas MMP 3 was over-expressed in response to Ad-TbetaRIIDeltacyt in HSCs. Moreover, zymography assays corroborated MMP-2 and MMP-3 changes in activity. Surprisingly, anti-TGF-beta molecular intervention increased nuclear SnoN in HSCs. In vivo, Ad TbetaRIIDeltacyt reduced liver fibrosis, increased nuclear SnoN in sinusoidal cells, and also produced significant suppression in collagen alpha1 (I), TGF beta1, PAI-1, MMP-2 and over-expression in MMP-3 in thioacetamide-intoxicated animals. CONCLUSIONS: The results obtained in the present study suggest that the molecular mechanism for the blocking effects of Ad-TbetaRIIDeltacyt in TGF-beta signaling acts via up-regulation of the transcriptional repressor SnoN, which antagonizes TGF-beta signaling (TGF-beta/Smad-pathway inhibitor). Consequently, profibrogenic genes are down-regulated. PMID- 19189316 TI - Post-myocardial infarction intraventricular conduction defects and B-type natriuretic peptide levels. AB - BACKGROUND: Levels of B-type natriuretic peptide (BNP), a significant marker of left ventricular (LV) dysfunction, may provide information on the severity of heart failure in patients with intraventricular conduction defects (IVCD).The aim of this study was to measure serum BNP levels in post-myocardial infarction (MI) patients with IVCD. METHODS: In 2004-2005 during standard follow-up at 6 mo after acute MI in 158 subjects, BNP was measured using an immunoenzymatic method. Of them, 126 patients had IVCD: 31, left bundle branch block (LBBB); 36, right bundle branch block (RBBB); 30, left anterior hemiblock (LAH); and 29, left posterior hemiblock (LPH). Thirty two patients with a previous MI, but without IVCD, served as controls, whereas 15 healthy subjects entered the comparative group. RESULTS: In post-MI patients with IVCD, the mean BNP level was significantly higher than in their counterparts without IVCD (p < 0.001) and in healthy subjects (p < 0.001). The highest BNP level was found in patients with LBBB and RBBB. In patients with LAH and LPH, levels of BNP were insignificantly higher than in patients without IVCD. CONCLUSIONS: Significantly higher BNP levels in patients with previous MI and IVCD suggest more severe heart failure than in their counterparts without IVCD with comparable LV ejection fraction. PMID- 19189317 TI - Implications of an avian-style respiratory system for gigantism in sauropod dinosaurs. AB - In light of evidence for avian-like lungs in saurischian dinosaurs, the physiological implications of cross-current gas exchange and voluminous, highly heterogeneous lungs for sauropod gigantism are critically examined. At 12 ton the predicted body temperature and metabolic rate of a growing sauropod would be similar to that of a bird scaled to the same body weight, but would increase exponentially as body mass increases. Although avian-like lung structure would be consistent with either a tachymetabolic-endothermic or a bradymetabolic gigantothermic model, increasing body temperature requires adjustments to avoid overheating. We suggest that a unique sauropod structure/function unit facilitated the evolution of gigantism. This unit consisted of (1) a reduction in metabolic rate below that predicted by the body temperature, akin to thermal adaptation as seen in extant squamates, (2) presence of air-filled diverticula in the long neck and in the visceral cavity, and (3) low activity of respiratory muscles coupled with the high efficiency of cross-current gas exchange. PMID- 19189318 TI - Synthesis of 5-(4'-carboxyphenyl)-10,15,20-tris-(4 pyridyl)-porphyrin and its peptidyl phosphonate derivatives. AB - The synthesis and characterization of three new 4-pyridyl porphyrin-peptidyl phosphonate compounds, containing a diphenyl 3-pyridylmethyl-phosphonate moiety, is described in this article. Nitrogen atoms in the pyridine rings of the obtained compounds were alkylated using methyl iodide, to give additional three, water soluble derivatives of these peptidyl-porphyrin conjugates. All the synthesized compounds could serve as potential photosensitizers for the photodynamic therapy (PDT) method of tumor therapy and displayed activity as inhibitors of aminopeptidase N. PMID- 19189320 TI - Religious beliefs, social support, self-efficacy and adjustment to cancer. AB - PURPOSE: Religious beliefs have received relatively little attention in research on coping with cancer. In this study, the relationship of religious beliefs and perceived social support with adjustment to cancer was studied in a coping model that included self-efficacy for coping as a mediator. Of particular interest was the relationship between religious beliefs and social support. METHOD: Data were collected from 164 in-treatment cancer patients. They completed measures of religious beliefs, social support, physical functioning, self-efficacy for coping, and adjustment. A model comparison approach was used to assess the fit of models that included or excluded the contribution of religious beliefs while testing the relationship between religious beliefs and social support. RESULTS: Religious beliefs were more strongly connected to perceived social support than with other constructs. Importantly, a coping model that included religious beliefs fit the data significantly better than a model without paths related to religious beliefs. Self-efficacy partially mediated the relation of age, physical functioning, and perceived support to adjustment, but not religious beliefs. DISCUSSION: Religious beliefs may not directly affect self-efficacy and adjustment; however, cancer patients who have religious beliefs may experience an enhanced sense of social support from a community with whom they share those beliefs. PMID- 19189319 TI - Marital adjustment, satisfaction and dissolution among hematopoietic stem cell transplant patients and spouses: a prospective, five-year longitudinal investigation. AB - OBJECTIVE: To examine the trajectory of marital adjustment, satisfaction and dissolution among 121 hematopoietic stem cell transplant (HSCT) couples-as a function of role (patient or spouse) and gender. METHODS: Prospective, longitudinal design. Participants completed the Dyadic Adjustment Scale at six time points: pre-transplant (4-7 days after beginning medical workup prior to starting transplant), 6 months post-transplant, and 1, 2, 3 and 5 years post transplant. They also reported on marital status over time. RESULTS: Participants ranged in age from 23-68 (52% female patients and 48% female spouses). Ninety eight patients received an allogeneic transplant; 23 received an autologous transplant. Marital dissolution was uncommon (four divorces since the transplant among 55 participating 5-year survivors, 7%). Dyadic satisfaction was stable over time for both male and female patients and male spouses, but not for female spouses who reported reductions in satisfaction at all time points relative to baseline. CONCLUSION: Couples were by and large resilient. However, caregiver specific gender differences indicate an increased risk for relationship maladjustment and dissatisfaction among female spouses and ultimately, female spouse/male patient couples. PMID- 19189322 TI - Rational synthesis of magnetic thermosensitive microcontainers as targeting drug carriers. AB - A new approach to the fabrication of magnetic thermosensitive microcontainers of Fe(3)O(4) nanoparticles with poly(N-isopropylacrylamide) walls is presented. The microcontainers undergo a temperature-induced volume phase transition and present an impressive magnetic response. The microcontainers have a well-defined structure with a narrow size distribution. The wall thicknesses of the microcontainers can be controlled according to requirements. Compared to other preparation methods, the process is simple and reproducible. The magnetic saturation of these microcontainers is high enough to meet most requirements of bioapplications. To further investigate the potential application of these microcontainers, they are tested as drug carriers, with the drug loading and releasing processes carefully studied. The drug encapsulation efficiency and drug content in the carriers are pH-dependent, and the carriers have a maximal drug loading of about 50 wt% under alkaline conditions. The release of the drug from the microcontainers can be controlled by the environmental pH, temperature, and magnetic force. PMID- 19189321 TI - Nmp4/CIZ suppresses parathyroid hormone-induced increases in trabecular bone. AB - The nucleocytoplasmic shuttling transcription factor Nmp4/CIZ (nuclear matrix protein 4/cas interacting zinc finger protein) is a ubiquitously expressed protein that regulates both cytoplasmic and nuclear activities. In the nucleus, Nmp4/CIZ represses transcription of genes crucial to osteoblast differentiation and genes activated by various anabolic stimuli, including parathyroid hormone (PTH). We investigated the role of Nmp4/CIZ in the PTH-induced increase in bone by engineering mice with loss-of-function mutations in the Nmp4/CIZ gene, and treating 10-week-old female mice with anabolic doses of human PTH (1-34) at 30 microg/kg/day, 7 day/week, for 7 weeks or vehicle control. The untreated, baseline phenotype of the Nmp4-null mice between 8 and 16 weeks of age included a modest but significant increase in bone mineral density (BMD) and bone mineral content (BMC) compared to wild-type (WT) mice. Type I collagen mRNA expression was moderately elevated in the femurs of the Nmp4-null mice. The Nmp4 mutant alleles decreased body weight by 4% when expressed on a mixed background but the same alleles on a pure B6 background yielded a significant, 15% increase in body weight among the KO mice, compared to their WT controls. Hormone treatment equally enhanced BMD and BMC over vehicle-treated mice in both the WT and Nmp4 null groups but Nmp4-KO mice exhibited a significantly greater PTH-induced acquisition of femoral trabecular bone as compared to WT mice. These data support our hypothesis that Nmp4/CIZ is a transcriptional attenuator that suppresses osteoid synthesis and PTH-mediated acquisition of cancellous bone. J. Cell. Physiol. 219: 734-743, 2009. (c) 2009 Wiley-Liss, Inc. PMID- 19189323 TI - A systematic nomenclature for codifying engineered nanostructures. AB - Nanotechnology's growing applications are fueled by the synthesis and engineering of a myriad nanostructures, yet there is no systematic naming or classification scheme for such materials. This lack of a coherent nomenclature is confusing the interpretation of data sets and threatens to hamper the pace of progress and risk assessment. A systematic nomenclature that encodes the overall composition, size, shape, core and ligand chemistry, and solubility of nanostructures is presented. A typographic string of minimalist field codes facilitates digital archiving and searches for desired properties. This nomenclature system could also be used for nanomaterial hazard labeling. PMID- 19189324 TI - Large-scale ordered plastic nanopillars for quantitative live-cell imaging. PMID- 19189325 TI - Cytochrome C on silica nanoparticles: influence of nanoparticle size on protein structure, stability, and activity. AB - The structure, thermodynamic and kinetic stability, and activity of cytochrome c (cyt c) on silica nanoparticles (SNPs) of different sizes have been studied. Adsorption of cyt c onto larger SNPs results in both greater disruption of the cyt c global structure and more significant changes of the local heme microenvironment than upon adsorption onto smaller SNPs. The disruption of the heme microenvironment leads to a more solvent-accessible protein active site, as suggested by Soret circular dichroism spectroscopy and through an increase in peroxidase activity as a function of increased SNP size. Similarly, the stability of cyt c decreases more dramatically upon adsorption onto larger SNPs. These results are consistent with changes in protein-nanoparticle interactions that depend on the size or surface curvature of the supporting nanostructure. This study provides further fundamental insights into the effects of nanoscale surfaces on adsorbed protein structure and function. PMID- 19189326 TI - Three-dimensional interconnected silica nanotubes templated from hyperbranched nanowires. PMID- 19189327 TI - Aerobic synthesis of cu nanoplates with intense plasmon resonances. PMID- 19189328 TI - High-strength, high-toughness composite fibers by swelling Kevlar in nanotube suspensions. PMID- 19189329 TI - Synthesis of AuAg alloy nanoparticles from core/shell-structured Ag/Au. PMID- 19189330 TI - Laser emission from electrospun polymer nanofibers. PMID- 19189331 TI - Cobalt nanoparticle arrays made by templated solid-state dewetting. AB - Self-assembled cobalt particle arrays are formed by annealing, which cause agglomeration (dewetting) of thin Co films on oxidized silicon substrates that are topographically prepatterned with an array of 200-nm-period pits. The Co nanoparticle size and uniformity are related to the initial film thickness, annealing temperature, and template geometry. One particle per 200-nm-period pit is formed from a 15-nm film annealed at 850 degrees C; on a smooth substrate, the same annealing process forms particles with an average interparticle distance of 200 nm. Laser annealing enables templated dewetting of 5-nm-thick films to give one particle per pit. Although the as-deposited films exhibit a mixture of hexagonal close-packed and face-centered cubic (fcc) phases, the ordered cobalt particles are predominantly twinned fcc crystals with weak magnetic anisotropy. Templated dewetting is shown to provide a method for forming arrays of nanoparticles with well-controlled sizes and positions. PMID- 19189332 TI - Preparation of uniform microspheres using a simple fluidic device and their crystallization into close-packed lattices. PMID- 19189333 TI - A simple top-down/bottom-up approach to sectored, ordered arrays of nanoscopic elements using block copolymers. AB - A top-down/bottom-up approach is demonstrated by combining electron-beam (e-beam) lithography and a solvent annealing process. Micellar arrays of polystyrene-block poly(4-vinylpyridine) (PS-b-P4VP) with a high degree of lateral order can be produced on a surface where sectoring is defined by e-beam patterning. The e-beam is used to crosslink the block copolymer (BCP) film immediately after spin coating when the BCP is disordered or in a highly ordered solvent-annealed film. Any patterns can be written into the BCP by crosslinking. Upon exposure to a preferential solvent for the minor component block followed by drying, cylindrical nanopores are generated within the nonexposed areas by a surface reconstruction process, while, in the exposed areas, the films remain unchanged. Nickel nanodot arrays can be placed over selected areas on a surface by thermal evaporation and lift-off process. PMID- 19189334 TI - Removal of second branchial cleft cysts using a retroauricular approach. AB - BACKGROUND: Second branchial cleft cysts (also called cervical lymphoepithelial cysts) usually present in adulthood, and cosmesis of surgical access is an important issue. This study was performed to assess the feasibility and outcome of a retroauricular approach for removing second branchial cleft cysts. METHODS: Eight patients underwent removal of second branchial cleft cysts using a retroauricular approach. RESULTS: All 8 second branchial cleft cysts were removed successfully, and the wounds healed uneventfully. None of the patients suffered from weakness of the great auricular nerve. Marginal nerve palsy did not occur postsurgery. Average follow-up period was 14.5 months (range, 6-20 months) with no tumor recurrence. The incision scars were invisible. CONCLUSION: Second branchial cleft cyst resection using a retroauricular approach is a feasible method that provides an acceptable cosmetic outcome. PMID- 19189335 TI - Central adenoid cystic carcinoma of the mandible with odontogenic features: Report of a case. AB - BACKGROUND: Primary intraosseous salivary-type adenocarcinomas are rare neoplasms of uncertain histogenesis. The prevailing theories suggest origin from heterotopic salivary glands, odontogenic rests, or cystic epithelium. METHODS: A case of central adenoid cystic carcinoma is reported in a 53-year-old woman who presented with a painless swelling in the anterior segment of her lower jaw. Radiographic examination confirmed the presence of an expansile, radiolucent lesion within the mandible with a multilocular appearance. On light microscopy analysis, areas showing cribriform and tubular growth patterns; admixed cysts, some of which were characterized by the presence of localized plaque-like thickenings of their epithelial linings; and the formation of aberrant dental hard tissue were observed. RESULTS: The radiographic and histomorphologic findings highlight the potential misdiagnosis of this rare tumor. The findings also draw attention to an embryologic histogenetic concept for some central salivary neoplasms. CONCLUSION: The divergent salivary and odontogenic differentiation evident in this tumor signifies the pluripotential nature of derivatives of oral ectoderm. PMID- 19189336 TI - Anterolateral thigh free flap for tracheal reconstruction after parastomal recurrence. AB - BACKGROUND: Stomal recurrence after total laryngectomy is 1 of the most serious issues in head and neck surgery, both because of the complexity of its management and because of its morbidity. Prior to the introduction of free-tissue transfer, mediastinal tracheostomy has been the standard reconstructive procedure with high rate of complications. The ideal reconstructive solution to these problems must provide well-vascularized soft tissues that can cover the defect after resection and also allow suturing of the tracheal remnant to skin edges without tension. METHODS AND RESULTS: We describe a case of a 56-year-old man with stomal recurrence after total laryngectomy treated by the use of a tubed anterolateral thigh (ALT) flap to elongate the shortened trachea and simultaneously cover the cervical skin defect. CONCLUSIONS: The ALT can be accepted as an ideal free-flap choice for stomal recurrence, because it has maximal reconstructive capacity and produces minimal donor-site morbidity. PMID- 19189337 TI - Playing a brass instrument after total laryngectomy: a case report. AB - BACKGROUND: A brass instrument is a musical instrument in which the tone is produced by vibration of the lips as the player blows into a tubular resonator. The case of a professional brass player who continued his activity after total laryngectomy, with insertion of a voice prosthesis in a tracheoesophageal shunt, is reported. METHODS: A videoendoscopic and videofluoroscopic study of the patient during brass playing was conducted. RESULTS: A nonvibrating, open neoglottis during brass playing was found on videoendoscopy. Videofluorography revealed an enlarged hypopharynx, a thick neoglottis while playing at lower tone; at higher pitch the tongue base was retracted, the neoglottis was thin and stretched, and the subneoglottic area was extremely enlarged. CONCLUSION: The case reported shows that the insertion of a voice prosthesis in a tracheoesophageal shunt seems to create a regulating airflow system sufficiently advanced to play a brass instrument, further reducing the disability of laryngeal speakers. PMID- 19189338 TI - Advanced squamous cell carcinoma of the oropharynx: efficacy of positron emission tomography and computed tomography for determining primary tumor response during induction chemotherapy. AB - BACKGROUND: To evaluate the efficacy of fluorine-18-fluorodeoxyglucose emission tomography (FDG-PET) and CT versus endoscopy with biopsy under general anesthesia for estimating tumor volume reduction among patients treated with induction chemotherapy for advanced squamous cell carcinoma (SCC) of the oropharynx. METHODS: Twelve patients with oropharyngeal SCC nested in a phase II, induction chemoradiation, organ preservation trial (University of Michigan Cancer Center 9921) underwent tumor volume reduction estimation as assessed by FDG-PET, CT, and endoscopy with biopsy. RESULTS: In 9 of 12 patients, FDG-PET, CT, and endoscopy demonstrated agreement in estimation of tumor reduction. Two patients had discordant results, whereas 1 patient was inadequately evaluated with FDG-PET. The kappa value for PET versus endoscopy was 0.62, which is categorized as substantial agreement. The kappa value for CT versus endoscopy was 0.40, which is categorized as fair agreement. CONCLUSION: FDG-PET may be as efficacious as endoscopy with biopsy under general anesthesia for estimating tumor volume reduction with induction chemotherapy. PMID- 19189339 TI - Retropharyngeal nodal metastasis related to higher rate of distant metastasis in patients with N0 and N1 nasopharyngeal cancer. AB - BACKGROUND: Retropharyngeal lymph node (RLN) staging in nasopharyngeal carcinoma (NPC) can be controversial. METHODS: We retrospectively reviewed all patients with T(2-4), N(0-1) NPC treated between 1992 and 1994 to examine if RLN metastasis resulted in an increased incidence of distant metastases. RESULTS: Of the 667 patients with NPC, 395 had T(2-4), N(0-1) disease, 140 had N(0), and 255 had N(1). All had staging CT scans and were treated with radiotherapy. Median follow-up was 8.3 years. Seventy-four percent showed undifferentiated histology. In this cohort, 187 (47%) had RLN metastases. Multivariate analysis showed that RLN conferred a higher hazard for distant metastasis (p = .04). Using the Kaplan Meier method, patients with N(0) disease and RLN had a similar hazard for distant metastases as patients with N(1) disease when compared with patients with N(0) disease and without RLN. CONCLUSION: Patients with N(0) disease and RLN appear to share a similar prognosis to patients with N(1) disease. PMID- 19189340 TI - National Cancer Database report on cancer of the head and neck: 10-year update. AB - BACKGROUND: We sought to examine the current state of cancer care for head and neck tumors in the United States. We therefore performed a retrospective, longitudinal study of the approximately 822,000 head and neck cancer cases included in the National Cancer Data Base (NCDB) for 1990 through 2004, representing approximately 75% of the estimated incident diagnoses in the United States. METHODS: All cases of head and neck cancer diagnosed and reported to the NCDB during this interval were reviewed, and descriptive statistics, grouped by disease and host factors, were analyzed over time and compared with a prior similar analysis done 10 years ago. RESULTS: Although many similarities persist, several major changes in head and neck cancer have occurred, most notably (1) a decrease in the number of the older-aged patients who have mucosally derived squamous cell carcinomas coupled with an increase in the number of younger-aged patients who have thyroid-origin adenocarcinomas and (2) a decrease in the use of radiation therapy alone for treatment in favor of chemotherapy enhanced radiation therapy. CONCLUSION: Head and neck cancers include a heterogeneous group of tumors whose precise composition changes over time and whose therapy evolves as well. The NCDB is well suited to capture this information and provide both an analysis of the current state of cancer care for head and neck tumors and a longitudinal view over time. PMID- 19189341 TI - Usefulness of 201TL SPECT/CT relative to 18F-FDG PET/CT in detecting recurrent skull base nasopharyngeal carcinoma. AB - BACKGROUND: This study was designed to compare (201)Tl single photon emission computed tomography (SPECT)/CT with 18-fluoro-2-deoxyglucose ((18)F-FDG) PET/CT in diagnosing recurrent skull base nasopharyngeal carcinoma (NPC). METHODS: Twenty-seven patients were recruited. Both (201)Tl SPECT/CT and (18)F-FDG PET/CT for each patient were performed at least 4 months later after initial therapy. RESULTS: The sensitivity and specificity for (201)Tl SPECT/CT were 66.7% and 100%, and those for (18)F-FDG PET/CT were 86.7% and 75.0%. Lesion/background ratios were obtained for the 10 lesions that were both SPECT and PET true positive. For the 8 patients with recurrences in nasopharyngeal regions, PET lesion/background ratios were all higher than SPECT lesion/background ratios. For the 2 patients with intracranial metastases, SPECT lesion/background ratios were higher than PET lesion/background ratios. CONCLUSION: (201)Tl SPECT/CT is as effective as (18)F-FDG PET/CT in detecting recurrent NPC. For intracranial recurrence, (201)Tl SPECT, because of its high intracranial lesion/background ratio, is probably better than (18)F-FDG PET. PMID- 19189342 TI - Concentration dependent dual effect of thrombin in endothelial cells via Par-1 and Pi3 Kinase. AB - Disruption of endothelial barrier is a critical pathophysiological factor in inflammation. Thrombin exerts a variety of cellular effects including inflammation and apoptosis through activation of the protease activated receptors (PARs). The activation of PAR-1 by thrombin is known to have a bimodal effect in endothelial cell permeability with a low concentration (pM levels) eliciting a barrier protective and a high concentration (nM levels) eliciting a barrier disruptive response. It is not known whether this PAR-1-dependent activity of thrombin is a unique phenomenon specific for the in vitro assay or it is part of a general anti-inflammatory effect of low concentrations of thrombin that may have a physiological relevance. Here, we report that low concentrations of thrombin or of PAR-1 agonist peptide induced significant anti-inflammatory activities. However, relatively high concentration of thrombin or of PAR-1 agonist peptide showed pro-inflammatory activities. By using function-blocking anti-PAR-1 antibodies and PI3 kinase inhibitor, we show that the direct anti inflammatory effects of low concentrations of thrombin are dependent on the activation of PAR-1 and PI3 kinase. These results suggest a role for cross communication between PAR-1 activation and PI3 kinase pathway in mediating the cytoprotective effects of low concentrations of thrombin in the cytokine stimulated endothelial cells. J. Cell. Physiol. 219: 744-751, 2009. (c) 2009 Wiley-Liss, Inc. PMID- 19189343 TI - Identification of new trace triterpenoid saponins from the roots of Panax notoginseng by high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry. AB - Triterpenoid saponins are the major bioactive constituents of Panax notoginseng. In the study reported here, the fragmentation behavior of triterpenoid saponins from P. notoginseng was investigated by electrospray ionization tandem mass spectrometry (ESI-MS(n))and high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HPLC/ESI-MS(n)). Analyses revealed that product ions from glycosidic and cross-ring cleavages can give a wealth of structural information regarding the nature of the aglycone, sugar types, the sequence and linkage information of sugar units. It is noted that different glycosylation positions remarkably influenced the fragmentation behaviors, which could assist in the differentiation of saponin analogues. To rationalize this characteristic, the collision energy required for various glycosidic cleavages was investigated. According to the summarized fragmentation rules, identification of triterpenoid saponins from the roots of P. notoginseng could be fulfilled, even when reference standards were unavailable. Furthermore, minor and trace constituents were enriched and detected by eliminating the major constituents in one of the saponin fractions. As a result, a total of 151 saponins, including 56 new trace ones, were identified or tentatively characterized from saponin fractions based on their retention times, HPLC/HRMS, HPLC/ESI-MS(n) fragmentation behaviors and comparison with literature data. PMID- 19189344 TI - A novel approach to the dissection of the human knee. AB - The knee is one of the most frequently injured joints of the human body with injuries affecting the general population and the athletic population of many age groups. Dissection procedures for the knee joint typically do not allow unobstructed visualization of the anterior cruciate or posterior cruciate ligaments without sacrificing the collateral ligaments. In many cases, the relationships of the intraarticular structures are lost as dissection systematically removes superficial structures to gain access to deeper structures. The authors present an alternative technique for dissection of the human knee joint that allows maximal visualization of intraarticular structures such as the cruciate ligaments and menisci with minimal disturbance to the tibial and fibular collateral ligaments, thus preserving the relationships of the ligamentous and intraarticular structures. PMID- 19189345 TI - Lysophosphatidic acid-induced ERK activation and chemotaxis in MC3T3-E1 preosteoblasts are independent of EGF receptor transactivation. AB - Bone-forming osteoblasts and their progenitors are target cells for the lipid growth factor lysophosphatidic acid (LPA) which is produced by degranulating platelets at sites of tissue injury. LPA is a potent inducer of bone cell chemotaxis, proliferation and survival in vitro, and this lipid factor is an attractive candidate to facilitate preosteoblast migration during skeletal regeneration in vivo. In this study we sought to more clearly define the intracellular signaling pathways mediating the effects of LPA on bone cells. LPA treated MC3T3-E1 preosteoblastic cells exhibited a bimodal activation of extracellular signal-related kinase (ERK1/2) with maximal phosphorylation at 5 and 60 min. MEK1/2 activation was detected within 2.5 min of LPA exposure and remained elevated for at least an hour. ERK1/2 phosphorylation was not coupled to Ras activation or to LPA-induced elevations in cytosolic Ca(2+). While LPA exposure transactivates the EGF receptor in many cell types, LPA-stimulated ERK1/2 activation in MC3T3-E1 cells was unaffected by the inhibition of EGF receptor function. ERK isoforms can function as transcription factors and ERK1/2 rapidly accumulated in the nuclei of LPA-treated cells, a process that was blocked if ERK1/2 phosphorylation was prevented. Blocking ERK1/2 phosphorylation also led to significant decreases in LPA-induced MC3T3-E1 cell chemotaxis, while the inhibition of EGF receptor function had no effect on the stimulation of preosteoblast motility by LPA. Our results identify ERK1/2 activation as a mediator of LPA-stimulated MC3T3-E1 cell migration that may be relevant to preosteoblast motility and gene expression during bone repair in vivo. J. Cell. Physiol. 219: 716-723, 2009. (c) 2009 Wiley-Liss, Inc. PMID- 19189346 TI - The Case Anatomical Knowledge Index (CAKI): a novel method used to assess anatomy content in clinical cases. AB - There are concerns in the literature that the use of case-based teaching of anatomy could be compromising the depth and scope of anatomy learned by students in a problem-based learning curriculum. Poor selection of clinical cases that are used as vehicles for teaching/learning anatomy may be the root problem because some clinical cases do not provide enough opportunities to learn anatomy and are, therefore, inappropriate for case-based teaching. Although anatomy educators are expected to respond to the identified deficiencies of case-based anatomy teaching, making sure that students acquire sufficient anatomical knowledge to practice safely and successfully, there are no tools available that can help improve the selection of clinical cases for case-based teaching. The author proposes a composite index, which incorporates considerations of anatomical knowledge for evaluating clinical cases/conditions for suitability in case-based anatomy teaching. The development of the case anatomical knowledge index (CAKI) using a modified Guttman procedure is described. The scalability of the index was measured using the coefficient of reproducibility. A total of 47 clinicians participated in the validation activities that measured interrater and intraclass reliability. The CAKI was able to consistently discriminate between clinical cases/conditions with higher demand for anatomical knowledge than those with lower demand for anatomical knowledge. A review of the literature suggests that such an index has not been previously reported. Given the concerns about the depth and scope of anatomy learning in case-based teaching, these findings have international relevance. PMID- 19189347 TI - Clay modeling as a method to learn human muscles: A community college study. AB - The efficacy of clay modeling compared with cat dissection for human muscle identification was examined over two semesters at LaGuardia Community College in Queens, NY. The 181 students in 10 sections in this study were randomly distributed into control (cat dissection) and experimental (clay modeling) groups, and the results of the muscle practical examination were analyzed. The clay-modeling group was significantly better at identifying human muscles on human models than the cat-dissection group, and was as good at identifying muscles on their self-made clay mannequins as the cat-dissection group was at identifying cat muscle on their specimens. This study demonstrated that clay modeling is more effective than cat dissection for learning human muscles at the community college level. PMID- 19189348 TI - 'Spice' and other herbal blends: harmless incense or cannabinoid designer drugs? PMID- 19189349 TI - Role of surgical outcome as prognostic factor in advanced epithelial ovarian cancer: a combined exploratory analysis of 3 prospectively randomized phase 3 multicenter trials: by the Arbeitsgemeinschaft Gynaekologische Onkologie Studiengruppe Ovarialkarzinom (AGO-OVAR) and the Groupe d'Investigateurs Nationaux Pour les Etudes des Cancers de l'Ovaire (GINECO). AB - BACKGROUND: Primary surgery followed by platinum-taxane based chemotherapy has been the standard therapy in advanced ovarian cancer. However, the prognostic role of complete and so-called optimal and suboptimal debulking and its interaction with biological factors has not been not fully defined. METHODS: Exploratory analysis was conducted of 3 prospective randomized trials (AGO-OVAR 3, 5, and 7) investigating platinum-taxane based chemotherapy regimens in advanced ovarian cancer conducted between 1995 and 2002. RESULTS: A total of 3126 patients were analyzed. Approximately one-third each fulfilled criteria for complete resection (group A), small residual tumor burden of 1-10 mm (group B), or macroscopic residual disease exceeding 1 cm in diameter (group C). Multivariate analysis showed improved progression-free and overall survival for group A with complete resection compared with groups B or C (P<.0001). The impact of so-called optimal debulking as in group B showed a smaller prognostic impact compared with group C. Further independent prognostic factors for overall survival were age, performance status, grade, FIGO stage, and histology, namely the mucinous subtype. An interaction between residual tumor and some biologic factors was demonstrated. CONCLUSIONS: The goal of primary surgery should be complete resection. The prognostic impact of tumor biology seemed to be partially overruled by residual tumor and further evaluation of biologic factors should stratify for residual tumor. PMID- 19189350 TI - Risk factors for serious injury in Finnish agriculture. AB - BACKGROUND: Previous studies indicate 20% of injuries represent 80% of injury costs in agriculture. To help prevent the most costly injuries, we aimed to identify characteristics and risk factors associated with serious injuries. METHODS: We analyzed insurance records of 93,550 self-employed Finnish farmers. We ranked injury causes by claim cost and used multiple logistic regressions to identify risk factors for (any) injury and serious injury (injuries exceeding claim costs of 2000 euros). RESULTS: A total of 5,507 compensated injuries occurred in 2002 (rate 5.9/100 person-years), and 1,167 or 21% of them (rate 1.25/100 person-years) were serious. The causes/sources resulting in highest average claim costs were motor vehicles; stairs, scaffoldings, and ladders; trailers and wagons; floors, walkways, and steps; other structures and obstacles; augers, mills, and grain handling equipment; horses; combines and harvesting equipment; tractor steps; and uneven and slippery terrain. Older age, male gender, higher income level, greater field size, residing on the farm, Finnish language (vs. Swedish), occupational health service (OHS) membership, and animal production were risk factors for injury. The risk factors for serious injury were similar; however, the effects of age, income level, and the raising of horses were more prominent. Language, residence, ownership status, and OHS membership were not risk factors for serious injury. CONCLUSIONS: Cost-effective prevention efforts should address the following risk factors: older age, male gender, larger income and operation size, livestock production (particularly dairy, swine, and horses), motor vehicle incidents, falls from elevation, and slips, trips and falls. PMID- 19189351 TI - Combined utilization of ion mobility and ultra-high-resolution mass spectrometry to identify multiply charged constituents in natural organic matter. AB - Natural organic matter as complex biogeochemical non-repetitive material was investigated with ion mobility mass spectrometry (IMS) and ultra-high-resolution Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS) approaches in order to unravel the existence of multiply charged state constituents. Hereby we describe and assign the potential molecular formulae of these doubly charged species, derived from FTICR-MS, and the existence of these species was confirmed via IMS. The parallel application of these powerful techniques enabled the boundaries of the understanding of natural organic matter to be pushed further. PMID- 19189352 TI - Field distribution in an electrospray ionization source determined by finite element method. AB - Three-dimensional computer models of electrospray ionization sources were constructed in COMSOL Multiphysics to solve the static electric fields using finite element methods. The magnitude of the electric field strength for onset of electrospray and optimum signal was calculated under various conditions. The modification of the electric field distribution in the ion source by an atmospheric pressure ion lens was also investigated by plotting the equipotential surfaces, electric field lines and trajectories of charged droplets. Both the calculated and the experimental results demonstrate that the changes in the ion signal detected by the mass spectrometer are attributable to the focusing effect of the ion lens when appropriate voltages are applied on the sprayer and ion lens. The optimum signal was found by setting the sprayer voltage from 3000 to 5000 V while scanning the ion lens voltage. The calculated strengths of the electric field at the sprayer tip for optimum signals are similar although the applied voltages at the sprayer and ion lens are significantly different. PMID- 19189353 TI - First application of the depth profile of silica species as a tracer by fast-atom bombardment mass spectrometry: investigation of the circulation of seawater and silica uptake by diatoms. AB - Silica, represented by SiO(2), is the general name for the compounds composed of Si, O and H with their derivative complexes. Silica forms various chemical species in aquatic solutions, such as a monomer (Si(OH)(3)O(-)), dimer (Si(2)(OH)(5)O(2) (-)), and others. These species are known to vary in their relative abundances in solution depending on the chemical and physical conditions. Silica species dissolved in seawater have been examined by fast-atom bombardment mass spectrometry (FAB-MS) to elucidate the behavior of silica and its circulation as a novel tracer reflecting the chemical and physical conditions of seawater and the bioactivity of diatoms, which take up silica. In the seawater of Tokyo Bay, silica species such as [Si(OH)(2)O(2)Na](-) ([monomer-Na](-)), [Si(2)(OH)(5)O(2)](-) ([dimer](-)), [Si(2)(OH)(4)O(3)Na](-) ([dimer-Na](-)), [Si(4)(OH)(7)O(5)](-) ([cyclic tetramer](-)), [Si(4)(OH)(6)O(6)Na](-) ([cyclic tetramer-Na](-)), [Si(4)(OH)(9)O(4)](-) ([linear tetramer](-)) and [Si(4)(OH)(8)O(5)Na](-) ([linear tetramer-Na](-)) were observed and assigned by FAB-MS. To investigate the suitability of silica species as a tracer, the relative peak intensity ratios of silica species observed in the mass spectra, i.e. the profiles of the ratio of the linear tetramer to the cyclic tetramer (m/z 329/311) and the ratio of the dimer to the cyclic tetramer (m/z 173/311) against depth, were examined to determine the annual changes and reproducibility of the depth profiles. In particular, the depth profile of the relative ratio of the linear tetramer to the cyclic tetramer, 329/311, exhibits critical changes depending on the seawater budget. These changes in the relative ratios were identified by an experiment involving a simple sodium chloride solution system. Our measurement is expected to elucidate the dynamics of silica and its role as 'food' for diatoms, and we showed that speciation using mass spectrometry is a powerful tool for examining elemental behavior in nature and environmental changes. Our results suggest that a silica tracer is useful for investigating the behavior of seawater in small coastal regions and the uptake of silica by diatoms. PMID- 19189355 TI - Toward volatile and nonvolatile molecular memories: fluorescence switching based on fluoride-triggered interconversion of simple porphyrin derivatives. AB - Written in the gel: A fluoride-writable memory system has been demonstrated by using two porphyrin derivatives. Structural modification enabled both volatile and nonvolatile modes. For both porphyrins, fluorescence is quenched by addition of F(-) with only one of the derivatives returning to its initial fluorescent state following removal of F(-) (see scheme). Nonvolatile behaviour is based on anion-induced conversion from porphyrin to oxoporphyrinogen. PMID- 19189356 TI - Highly sensitive magnetic effects induced by hydrogen-bonding interactions in a high-spin metallosupramolecular Fe(4) (II) [2x2] grid-type complex. AB - A sensitive magnetic nanoprobe: Hydrogen-bonding interactions are reflected with great sensitivity in the (1)H NMR spectra of a high-spin multinuclear Fe(4) (II) [2x2] grid-type complex (see scheme) and the measured shifts can be used to evaluate the hydrogen-bond donating ability. The grid complex also represents a prototype of a very sensitive magnetic nanoreceptor for the detection of very small changes around a magnetic center. PMID- 19189357 TI - Development of tau aggregation inhibitors for Alzheimer's disease. AB - A variety of human diseases are suspected to be directly linked to protein misfolding. Highly organized protein aggregates, called amyloid fibrils, and aggregation intermediates are observed; these are considered to be mediators of cellular toxicity and thus attract a great deal of attention from investigators. Neurodegenerative pathologies such as Alzheimer's disease account for a major part of these protein misfolding diseases. The last decade has witnessed a renaissance of interest in inhibitors of tau aggregation as potential disease modifying drugs for Alzheimer's disease and other "tauopathies". The recent report of a phase II clinical trial with the tau aggregation inhibitor MTC could hold promise for the validation of the concept. This Review summarizes the available data concerning small-molecule inhibitors of tau aggregation from a medicinal chemistry point of view. PMID- 19189358 TI - Duplex stabilization and energy transfer in zipper porphyrin-DNA. AB - Zip it up: Attachment of porphyrins onto complementary DNA strands leads to zipper-porphyrin arrays and, in the presence of eleven modifications, an increase in the melting temperature of the duplex. Mixed zinc and free-base porphyrin arrays undergo energy transfer from the zinc porphyrin to the free-base porphyrin in the annealed duplex but not in the denatured form (see scheme), giving access to reversible formation of potential photonic wires. PMID- 19189359 TI - On the "gluing" effect of lithium: the lithium-driven assembly of circum arranged, edge-fused cyclopentadienyl lithium compounds and aza analogues. AB - Predictions (DFT/B3LYP calculations) are that cyclopentadienyl lithium edge-fused to [n]circulenes in a circum-like manner should self-assemble as rod-like, nanometer long, supersandwich compounds (see figure). On the contrary, triazolyl lithium analogues prefer to dimerize thereby giving rise to shell-like dimers of variable curvatures.This study is aimed at exploring M. Etter's (T. W. Panunto, Z. Urbanczyk-Lipkowska, R. Jonhson, M. Etter, J. Am. Chem. Soc. 1987, 109, 7786 7797) bottom-up approach towards the design of organic-solid sate materials. Guided by the electrostatic paste ("gluing") effect of lithium, we examined the self-assembly modes of a series of cyclopentadienyl lithium edge-fused to benzene, as well as to [n]circulenes, arranged in a circum-like manner by means of DFT/B3 LYP calculations. We have also examined the self-assembly modes of the analogous triazolyl derivatives. Generally, well defined trends have been found. Thus, whereas cylcopentadienyl derivatives 1, 3 and 5 tend to aggregate as rod like, nanometer-long, supersandwich compounds, the corresponding triazolyl analogues 2, 4 and 6 tend to dimerize thus giving rise to shell-like dimers of variable curvatures. In our view, M. Etter's bottom-up approach combined with high level calculations appears to have great potential for designing complex molecular architectures and nanostructures. Hopefully, these studies will spur the activity of synthetic and materials chemists. PMID- 19189360 TI - Synthesis and characterization of [Ar'GaC(Ph)CH]2 and K2[Ar'GaC(Ph)CH]2OEt2: from digallene to digallacyclohexadiene to digallatabenzene. AB - On a plane: Addition of PhCCH to Ar'GaGaAr' (Ar' = 2,6-(2,6 iPr(2)C(6)H(3))(2)C(6)H(3)) yielded the 1,4-digallacyclohexadiene [{Ar'GaC(Ph)CH}(2)] (1), which afforded the 1,4-digallatabenzene dianion [Ar'GaC(Ph)CH](2)(2-) (2) upon reduction with potassium (see scheme; C gray, Ga green, K blue). Structural parameters, DFT calculations, and (1)H NMR spectroscopy support aromatic character of 2. PMID- 19189361 TI - Insertion of amides into a polyoxometalate. AB - POM alert: The incorporation of an amide oxygen atom into the framework of the Dawson-type polyoxometalate (POM) cluster [P(2)V(3)W(15)O(62)](9-) (see picture) allows the communication of electronic effects between the organic and the inorganic parts of the molecule, including fine-tuning of the redox properties of the entire hybrid POM by the organic components, and transmission of the POM's electron-attracting properties to the organic moiety. PMID- 19189362 TI - Mapping the limits of substrate specificity of the adenylation domain of TycA. AB - The catalytic potential of tyrocidine synthetase 1 (TycA) was probed by the kinetic characterization of its adenylation activity. We observed reactions with 30 substrates, thus suggesting some substrate promiscuity. However, although the TycA adenylation (A) domain was able to accommodate alternative substrates, their k(cat)/K(M) values ranged over six orders of magnitude. A comparison of the activities allowed the systematic mapping of the substrate specificity determinants of the TycA A-domain. Hydrophobicity plays a major role in the recognition of substrate analogues but can be combined with shape complementarity, conferring higher activity, and/or steric exclusion, leading to substantial discrimination against larger substrates. A comparison of the k(cat)/K(M) values of the TycA A-domain and phenylalanyl-tRNA synthetase showed that the level of discrimination was comparable in the two enzymes for the adenylation reaction and suggested that TycA was also subjected to high selective pressure. The specificity patterns observed and the quantification of alternative activities provide a basis for exploring possible paths for the future directed evolution of A-domain specificity. PMID- 19189363 TI - Cooperative effects on radical recombination in CYP3A4-catalyzed oxidation of the radical clock beta-thujone. AB - The timing of the beta-thujone radical clock (see scheme) can be specifically altered by an allosteric effector. Progesterone, a well-documented CYP3A4 allosteric effector, was found to increase the yield of the unrearranged, C4 derived product of beta-thujone oxidation at the expense of the combined yields of all the rearranged C4-oxidized metabolites. The results demonstrate that the apparent radical recombination rate in the CYP3A4 hydroxylation of beta-thujone is accelerated by the progesterone heterotropic cooperativity. PMID- 19189364 TI - Differential regulation of protein subdomain activity with caged bivalent ligands. AB - Subtle change: Spatiotemporal modulation of individual protein subdomains with light as the trigger signal becomes possible by using bivalent aptamers and introducing photolabile "caging groups" to switch individual aptamer modules ON or OFF differentially. To the best of our knowledge, this is the first study to show that it is possible to modulate individual domain activity in aptamers, and thus also domain activity in proteins, with light. PMID- 19189365 TI - Is it possible to dope single-walled carbon nanotubes and graphene with sulfur? AB - Herein, we investigate sulfur substitutional defects in single-walled carbon nanotubes (SWCNTs) and graphene by using first-principles calculations. The estimated formation energies for the (3,3), (5,5), and (10,0) SWCNTs and graphene lie between 0.9 and 3.8 eV, at sulfur concentrations of 1.7-4 atom %. Thus, from a thermodynamic standpoint, sulfur doping is not difficult. Indeed, these values can be compared with that of 0.7 eV obtained for a nitrogen-doped (5,5) SWCNT. We suggest that it may be possible to introduce sulfur into the SWCNT framework by employing sulfur-containing heterocycles. Our simulations indicate that sulfur doping can modify the electronic structure of the SWCNTs and graphene, depending on the sulfur content. In the case of graphene, sulfur doping can induce different effects: the doped sheet can be a small-band-gap semiconductor, or it can have better metallic properties than the pristine sheet. Thus, S-doped graphene may be a smart choice for constructing nanoelectronic devices, since it is possible to modulate the electronic properties of the sheet by adjusting the amount of sulfur introduced. Different synthetic routes to produce sulfur-doped graphene are discussed. PMID- 19189366 TI - Glycopeptide dendrimers with high affinity for the fucose-binding lectin LecB from Pseudomonas aeruginosa. AB - The fucose-specific lectin LecB is implicated in tissue binding and biofilm formation by the opportunistic pathogen Pseudomonas aeruginosa, which causes severe respiratory tract infections mainly in immunocompromised patients or cancer patients undergoing chemotherapy. With a view to developing multivalent LecB inhibitors as novel antibacterial agents, a combinatorial library containing 15 625 tetravalent C-fucosyl peptide dendrimers with the basic structure (CFuc X(6)X(5)X(4))(4)(LysX(3)X(2)X(1))(2)LysIleHisNH(2) (CFuc=alpha-L-fucosyl acetic acid, X(1-6)=amino acids, Lys=lysine branching) was screened for lectin binding using on-bead binding assays. Ten tetravalent and three octavalent dendrimers derived from the identified sequences were prepared by solid-phase peptide synthesis (SPPS), cleaved from the resin, and purified by preparative HPLC. Relative affinities of these soluble ligands to LecB were determined by an enzyme linked lectin assay (ELLA). Strong binding was observed for tetravalent and octavalent ligands, with up to 440-fold enhancement in potency over fucose for the octavalent cationic dendrimer 2G3 (CFuc LysPro)(8)(LysLeuPhe)(4)(LysLysIle)(2)LysHisIleNH(2)). Mono- and divalent controls showed affinities similar to fucose, highlighting the importance of multivalency for binding. Docking studies showed that the C-fucosyl group of the dendrimers can adopt the same binding mode as fucose itself, with the peptide arms protruding from the binding pocket and establishing specific contacts with the lectin. PMID- 19189367 TI - Stability of dimeric interface in banana lectin: Insight from molecular dynamics simulations. AB - Banana lectin (Banlec) is a homodimeric non-glycosylated protein. It exhibits the beta-prism I structure. High-temperature molecular dynamics simulations have been utilized to monitor and understand early stages of thermally induced unfolding of Banlec. The present study elucidates the behavior of the dimeric protein at four different temperatures and compares the structural and conformational changes to that of the minimized crystal structure. The process of unfolding was monitored by following the radius of gyration, the rms deviation of each residue, change in relative solvent accessibility and the pattern of inter- and intra-subunit interactions. The overall study demonstrates that the Banlec dimer is a highly stable structure, and the stability is mostly contributed by interfacial interactions. It maintains its overall conformation during high-temperature (400 500 K) simulations, with only the unstructured loop regions acquiring greater momentum under such condition. Nevertheless, at still higher temperatures (600 K) the tertiary structure is gradually lost which later extends to loss of secondary structural elements. The pattern of hydrogen bonding within the subunit and at the interface across different stages has been analyzed and has provided rationale for its intrinsic high stability. PMID- 19189368 TI - Factors related to the presentation of thin and thick nodular melanoma from a population-based cancer registry in Queensland Australia. AB - BACKGROUND: Worldwide, the incidence of thick melanoma has not declined, and the nodular melanoma (NM) subtype accounts for nearly 40% of newly diagnosed thick melanoma. To assess differences between patients with thin (or=2.01 mm) nodular melanoma, the authors evaluated factors such as demographics, melanoma detection patterns, tumor visibility, and physician screening for NM alone and compared clinical presentation and anatomic location of NM with superficial spreading melanoma (SSM). METHODS: The authors used data from a large population-based study of Queensland (Australia) residents diagnosed with melanoma. Queensland residents aged 20 to 75 years with histologically confirmed first primary invasive cutaneous melanoma were eligible for the study, and all questionnaires were conducted by telephone (response rate, 77.9%). RESULTS: During this 4-year period, 369 patients with nodular melanoma were interviewed, of whom 56.7% were diagnosed with tumors 2.00 mm). CONCLUSIONS: Awareness of factors related to earlier detection of potentially fatal nodular melanomas, including the benefits of a physician examination, should be useful in enhancing public and professional education strategies. Particular awareness of clinical warning signs associated with thin nodular melanoma should allow for more prompt diagnosis and treatment of this subtype. PMID- 19189369 TI - The Breast and Cervical Cancer Prevention and Treatment Act in Georgia: effects on time to Medicaid enrollment. AB - BACKGROUND: Implementation of the Breast and Cervical Cancer Prevention and Treatment Act of 2000 (BCCPTA) allowed states to extend Medicaid to any woman aged <65 without insurance screened and found to need treatment either for breast or cervical cancer or for a precancerous cervical condition through the National Breast and Cervical Cancer Early Detection Program (NBCCEDP) or in Georgia, other provider sites. METHODS: The authors used linked Georgia Comprehensive Cancer Registry (GCCR) and Medicaid data to test the: 1) likelihood of Medicaid enrollment in a given month and 2) time-to-enrollment (months) for those eventually enrolling. The authors used difference-in-differences analysis to estimate the effects of BCCPTA for breast or cervical cancer cases relative to a control group of women with other cancers. The authors controlled for sociodemographics, stage at diagnosis, year of diagnosis, and county level factors related to insurance levels in the area. RESULTS: Compared with the control cancer group, the hazard ratio of Medicaid enrollment for women with breast and cervical cancers increased post- vs pre-BCCPTA implementation. The estimated effect of this increase was that out of every 1000 women with breast cancer, BCCPTA led to 1.7 more (from 2.8 to 4.5 per month) enrolling in Medicaid. The results for women with local or later stages of cervical cancer indicated that of 1000 women with these cancers, the number enrolling in a given month increased by 3.4 due to BCCPTA. Results on time-to-enrollment indicated that the time between cancer diagnosis and enrollment was shortened by 7 to 8 months. CONCLUSIONS: The Georgia Medicaid program, in response to national legislation, increased the probability of women enrolling in Medicaid earlier and in turn, likely increased their cancer treatment options. PMID- 19189370 TI - Questions remain about the human papillomavirus vaccine. PMID- 19189371 TI - Association of progression-free survival, overall survival, and patient-reported outcomes by skin toxicity and KRAS status in patients receiving panitumumab monotherapy. AB - BACKGROUND: The authors explored the association of skin toxicity (ST) severity as measured by patient-reported ST and Common Terminology Criteria for Adverse Events (CTCAE) grading with efficacy of panitumumab, a fully human antiepidermal growth factor receptor antibody, from a phase 3 metastatic colorectal cancer (CRC) trial. METHODS: Patients were randomized to panitumumab plus best supportive care (BSC) vs BSC alone. ST by modified National Cancer Institute CTCAE v3.0 and modified Dermatology Life Quality Index (mDLQI), health-related quality of life (HRQOL), and CRC symptoms were measured. ST was analyzed using a landmark approach. Associations by KRAS mutational status were also assessed. RESULTS: Of 463 patients, 208 of 231 (90%) panitumumab patients and 184 of 232 (79%) BSC patients had > or = 1 postbaseline patient-reported outcome (PRO) assessment. Panitumumab patients with more severe ST had significantly longer overall survival (OS) (grade 2-4:grade 1; hazard ratio, 0.60; P = .0033). Lower mDLQI scores (< 67; more bothersome ST) were associated with longer OS (Cox model, P < .0001). Similar results were observed with progression-free survival (PFS). An inverse relation between mDLQI and HRQOL scores was observed, suggesting that ST bother correlated with better HRQOL. KRAS and PRO data were available in 363 patients (188 panitumumab; 175 BSC). Longer OS was associated with lower mDLQI scores, regardless of KRAS status. Longer PFS was associated with more severe ST (lower mDLQI scores and higher CTCAE grade ST) in patients with wild-type (WT) KRAS tumors, but not in patients with mutant KRAS tumors. CONCLUSIONS: More severe ST, by both clinical grading and PRO, is associated with better CRC symptoms and HRQOL and with longer OS and PFS among panitumumab treated patients. The associations for PFS were more pronounced in patients with WT KRAS tumors. PMID- 19189372 TI - In situ transmission electron microscopy and ion irradiation of ferritic materials. AB - The intermediate voltage electron microscope-tandem user facility in the Electron Microscopy Center at Argonne National Laboratory is described. The primary purpose of this facility is electron microscopy with in situ ion irradiation at controlled sample temperatures. To illustrate its capabilities and advantages a few results of two outside user projects are presented. The motion of dislocation loops formed during ion irradiation is illustrated in video data that reveals a striking reduction of motion in Fe-8%Cr over that in pure Fe. The development of extended defect structure is then shown to depend on this motion and the influence of nearby surfaces in the transmission electron microscopy thin samples. In a second project, the damage microstructure is followed to high dose (200 dpa) in an oxide dispersion strengthened ferritic alloy at 500 degrees C, and found to be qualitatively similar to that observed in the same alloy neutron irradiated at 420 degrees C. PMID- 19189373 TI - Puckering transitions of pseudoproline residues. AB - The puckering transitions of pesudoprolines such as oxazolidine and thiazolidine residues (Oxa and Thz dipeptides) with trans and cis prolyl peptide bonds were explored by optimizations along the endocyclic torsion angle chi(1) using quantum chemical methods in the gas phase and in water. The overall shapes of the potential energy surfaces for Oxa and Thz dipeptides in the gas phase and in water are similar to those for the Pro dipeptide, although there are some differences in relative stabilities of local minima and in barriers to puckering transition. On the whole, the barriers to puckering transition for Oxa and Thz dipeptides are computed to be 0.8-3.2 kcal/mol at the B3LYP/6-311++G(d,p) level in the gas phase and in water, which are lower by 0.5-1.9 kcal/mol than those for the Pro dipeptide. The n --> sigma* interactions for the delocalization of the lone pair of the prolyl amide nitrogen into the antibonding orbitals that are anti to the lone pair appear to play a role in stabilizing the nonplanar puckered transition states over the corresponding planar structures. The calculated barriers indicate that the down-to-up puckering transition can proceed in the orders Pro < Oxa < Thz in the gas phase and Pro approximately Oxa < Thz in water. (c) 2009 Wiley Periodicals, Inc. Biopolymers 91: 444-455, 2009. PMID- 19189374 TI - Rationally designed dehydroalanine (DeltaAla)-containing peptides inhibit amyloid beta (Abeta) peptide aggregation. AB - Among the pathological hallmarks of Alzheimer's disease (AD) is the deposition of amyloid-beta (Abeta) peptides, primarily Abeta (1-40) and Abeta (1-42), in the brain as senile plaques. A large body of evidence suggests that cognitive decline and dementia in AD patients arise from the formation of various aggregated forms of Abeta, including oligomers, protofibrils and fibrils. Hence, there is increasing interest in designing molecular agents that can impede the aggregation process and that can lead to the development of therapeutically viable compounds. Here, we demonstrate the ability of the specifically designed alpha,beta dehydroalanine (DeltaAla)-containing peptides P1 (K-L-V-F-DeltaA-I-DeltaA) and P2 (K-F-DeltaA-DeltaA-DeltaA-F) to inhibit Abeta (1-42) aggregation. The mechanism of interaction of the two peptides with Abeta (1-42) seemed to be different and distinct. Overall, the data reveal a novel application of DeltaAla-containing peptides as tools to disrupt Abeta aggregation that may lead to the development of anti-amyloid therapies not only for AD but also for many other protein misfolding diseases. (c) 2009 Wiley Periodicals, Inc. Biopolymers 91: 456-465, 2009. PMID- 19189375 TI - The (1)H, (13)C, (15)N resonance assignment, solution structure, and residue level stability of eosinophil cationic protein/RNase 3 determined by NMR spectroscopy. AB - Eosinophil cationic protein (ECP)/human RNase 3, a member of the RNase A family, is a remarkably cytotoxic protein implicated in asthma and allergies. These activities are probably due to ECP's ability to interact with and disrupt membranes and depend on two Trp, 19 Arg, and possibly an extremely high conformational stability. Here, we have used NMR spectroscopy to assign essentially all (1)H, (15)N, and backbone (13)C resonances, to solve the 3D structure in aqueous solution and to quantify its residue-level stability. The NMR solution structure was determined on the basis of 2316 distance constraints and is well-defined (backbone RMSD = 0.81 A). The N-terminus and the loop composed of residues 114-123 are relatively well-ordered; in contrast, conformational diversity is observed for the loop segments 17-22, 65-68, and 92 95 and most exposed sidechains. The side chain NH groups of the two Trp and 19 Arg showed no significant protection against hydrogen/deuterium exchange. The most protected NH groups belong to the first and last two beta-strands, and curiously, the first alpha-helix. Analysis of their exchange rates reveals a strikingly high global stability of 11.8 kcal/mol. This value and other stability measurements are used to better quantify ECP's unfolding thermodynamics. PMID- 19189376 TI - Synthesis of quadruplex-forming tetra-end-linked oligonucleotides: effects of the linker size on quadruplex topology and stability. AB - G-quadruplexes are characteristic structural arrangements of guanine-rich DNA sequences that abound in regions with relevant biological significance. These structures are highly polymorphic differing in the number and polarity of the strands, loop composition, and conformation. Furthermore, the cation species present in solution strongly influence the topology of the G-quadruplexes. Recently, we reported the synthesis and structural studies of new G-quadruplex forming oligodeoxynucleotides (ODNs) in which the 3'- and/or the 5'-ends of four ODN strands are linked together by a non-nucleotidic tetra-end-linker (TEL). These TEL-ODN analogs having the sequence TGGGGT are able to form parallel G quadruplexes characterized by a remarkable high thermal stability. We report here an investigation about the influence of the reduction of the TEL size on the molecularity, topology, and stability of the resulting TEL-G-quadruplexes using a combination of circular dichroism (CD), CD melting, (1)H NMR spectroscopy, gel electrophoresis, and molecular modeling data. We found that all TEL-(TGGGGT)(4) analogs, regardless the TEL size and the structural orientation of the ODN branches, formed parallel TEL-G-quadruplexes. The molecular modeling studies appear to be consistent with the experimental CD and NMR data revealing that the G-quadruplexes formed by TEL-ODNs having the longer TEL (L1-4) are more stable than the corresponding G-quadruplexes having the shorter TEL (S1-4). The relative stability of S1-4 was also reported. (c) 2009 Wiley Periodicals, Inc. Biopolymers 91: 466-477, 2009. PMID- 19189377 TI - Analysis of functional divergence within two structurally related glycoside hydrolase families. AB - Two glycoside hydrolase (GH) families were analyzed to detect the presence of functional divergence using the program DIVERGE. These two families, GH7 and GH16, each contain members related by amino acid sequence similarity, retaining hydrolytic mechanisms, and catalytic residue identity. GH7 and GH16 comprise GH Clan B, with a shared beta-jelly roll topology and mechanism. GH7 contains fungal cellobiohydrolases and endoglucanases and is divided into five main subfamilies, four of the former and one of the latter. Cluster comparisons between three of the cellobiohydrolase subfamilies and the endoglucanase subfamily identified specific amino acid residues that play a role in the functional divergence between the two enzyme types. GH16 contains subfamilies of bacterial agarases, xyloglucosyl transferases, 1,3-beta-D-glucanases, lichenases, and other enzymes with various substrate specificities and product profiles. Four cluster comparisons between these four main subfamilies again have identified amino acid residues involved in functional divergence between the subfamilies. (c) 2009 Wiley Periodicals, Inc. Biopolymers 91: 478-495, 2009. PMID- 19189378 TI - On the use of ESI-QqTOF-MS/MS for the comparative sequencing of nucleic acids. AB - The usability of a quadrupole-quadrupole-time-of-flight (QqTOF) instrument for the tandem mass spectrometric sequencing of oligodeoxynuleotides was investigated. The sample set consisted of 21 synthetic oligodeoxynucleotides ranging in length from 5 to 42 nucleotides. The sequences were randomly selected. For the majority of tested oligonucleotides, two or three different charge states were selected as precursor ions. Each precursor ion was fragmented applying several different collision voltages. Overall 282 fragment ion mass spectra were acquired. Computer-aided interpretation of fragment ion mass spectra was accomplished with a recently introduced comparative sequencing algorithm (COMPAS). The applied version of COMPAS was specifically optimized for the interpretation of information-rich spectra obtained on the QqTOF. Sequences of oligodeoxynucleotides as large as 26-mers were correctly verified in >94% of cases (182 of 192 spectra acquired). Fragment ion mass spectra of larger oligonucleotides were not specific enough for sequencing. Because of the occurrence of extensive internal fragmentation causing low sequence coverage paired with a high probability of assigning fragment ions to wrong sequences, tandem mass spectra obtained from oligonucleotides consisting of 30 and more nucleotides could not be used for sequence verification neither manually nor with COMPAS. (c) 2009 Wiley Periodicals, Inc. Biopolymers 91: 401-409, 2009. PMID- 19189379 TI - Rational shape engineering of the filamentous protein gamma prefoldin through incremental gene truncation. AB - An enticing possibility in nanotechnology is to use proteins as templates for the positioning of molecules in regular patterns with nanometer precision over large surface areas. However, the ability to redesign protein quaternary structure to construct new shapes remains underdeveloped. In the present work, we have engineered the dimensions of a filamentous protein, the gamma prefoldin (gamma PFD) from the hyperthermophile Methanocaldococcus jannaschii, and have achieved controllable attachment of filaments in a specific orientation on a carbon surface. Four different constructs of gamma PFD were generated in which the coiled coils extending from the association domain are progressively truncated. Three of the truncation constructs form well-defined filaments with predictable dimensions according to transmission electron microscopy. Two of these constructs had 2D persistence lengths similar to that of gamma PFD at 300-740 nm. In contrast, the 2D persistence length of the shortest truncation mutant was 3500 nm, indicating that the filament adsorbs along a different axis than the other constructs with its two rows of coiled coils facing out from the surface. The elastic moduli of the filaments range from 0.7-2.1 GPa, similar to rigid plastics and within the lower limit for proteins whose primary intermolecular interaction is hydrogen bonding. These results demonstrate a versatile approach for controlling the overall dimensions and surface orientation of protein filaments, and expand the toolbox by which to tune two overall dimensions in protein space for the creation of templated materials over a wide variety of conditions. (c) 2009 Wiley Periodicals, Inc. Biopolymers 91: 496-503, 2009. PMID- 19189380 TI - Continuity of aggression from childhood to early adulthood as a predictor of life outcomes: implications for the adolescent-limited and life-course-persistent models. AB - Using data from the Columbia County Longitudinal Study, a 40-year longitudinal study following an entire county's population of third-grade students from age 8 to 48, we examine questions about the long-term consequences of aggressive and antisocial behavior in childhood, adolescence, and young adulthood. We found moderate levels of continuity of aggression from age 8 to 48 both for males and for females. Contrary to what some have proposed, we found that continuity of aggressiveness is owing to not only the high-aggressive participants staying high but also owing to the low-aggressive participants staying low. Compared with life course-persistent low aggressives, we found that life-course-persistent high aggressives had consistently poorer outcomes across domains of life success, criminal behavior, and psychosocial functioning at age 48 (e.g., arrests, traffic violations, aggression toward spouse and divorces, depression, health, occupational and educational attainment). In contrast, adolescent-limited and child-limited aggressives did not differ from life-course-persistent low aggressives on the age 48 outcomes. Finally, the outcomes for late-onset (early adulthood) aggressives were also problematic in some domains though not as problematic as those for life-course-persistent aggressives. PMID- 19189381 TI - Arboreal habitat structure affects the performance and modes of locomotion of corn snakes (Elaphe guttata). AB - Arboreal environments pose many functional challenges for animal locomotion including fitting within narrow spaces, balancing on cylindrical surfaces, moving on inclines, and moving around branches that obstruct a straight path. Many species of snakes are arboreal and their elongate, flexible bodies appear well suited to meet many of these demands, but the effects of arboreal habitat structure on the locomotion of snakes are not well understood. We examined the effects of 108 combinations of surface shape (cylinder vs. rectangular tunnel), surface width, incline, and a row of pegs on the locomotion of corn snakes (Elaphe guttata). Pegs allowed the snakes to move on the widest and steepest surfaces that were impassable without pegs. Tunnels allowed the snakes to move on steeper inclines than cylinders with similar widths. The mode of locomotion changed with habitat structure. On surfaces without pegs, most snakes used two variants of concertina locomotion but always moved downhill using a controlled slide. Snakes used lateral undulation on most surfaces with pegs. The detrimental effects of increased uphill incline were greater than those of increased surface width on maximal velocity. Snakes moved faster in tunnels than on cylinders regardless of whether pegs were present. Depending on the surface width, the addition of pegs to horizontal cylinders and tunnels resulted in 8-24-fold and 1.3-3.1-fold increases in speed, respectively. Thus, pegs considerably enhanced the locomotor performance of snakes, although similar structures such as secondary branches seem likely to impede the locomotion of limbed arboreal animals. PMID- 19189382 TI - Molecular dynamics simulation of antimicrobial peptide arenicin-2: beta-hairpin stabilization by noncovalent interactions. AB - Arenicin-2 is a 21 residue antimicrobial cyclic peptide, possessing one disulphide bond between residues Cys(3) and Cys(20). NMR and CD studies suggested that the structure of arenicin-2 in water represented a well formed, but highly twisted beta-hairpin. To investigate the spatial arrangement of the peptide side chains and to get a clear view of its possible amphipathic properties we performed molecular dynamics in explicit water. Four independent trajectories, 50 ns in length, were produced, starting from various initial conformations or by applying different simulation conditions. Arenicin-2 retained its beta-hairpin structure during simulations, although the residues close to strand ends were found to escape from the ideal hairpin conformation. The type I' beta-turn connecting the two strands fluctuated between type IV and II' beta-turn. Conversely, the right-handed twist of the beta-hairpin was well conserved with average twist value 203 degrees +/- 19 degrees per eight residues. Several nonbonded interactions, like hydrophobic interactions between aliphatic side chains, cation/pi-aromatic interactions, CH...pi aromatic bond and water bridges, contributed to the hairpin stabilization. PMID- 19189383 TI - Spiral and square microstructured surfaces: the effect of the decreasing size of photo-immobilized hyaluronan domains on cell growth. AB - Spiral and squared micropatterned surfaces of decreasing dimensions were realized by photo-immobilizing a photoreactive hyaluronan (Hyal) derivative on silanized glass substrates. The microstructured surfaces were observed by atomic force microscope and scanning electron microscope. Scanning electron microscope analysis revealed the presence of a spiral (ranging from 100 microm down to 1 microm in the central part) and a square pattern consisting of a central square of 100 microm x 100 microm and squares of different dimensions decreasing from the centre to the edges of the micropatterned area (2 microm x 1 microm). Three cell types were tested on all the microstructured surfaces: human coronary artery endothelial cells (HCAEC), human dermal fibroblasts (C54), and NIH 3T3 fibroblasts. Cell adhesion analysis demonstrated that HCAEC and C54 did not adhere to the immobilized Hyal on silanized glass but adapted their shape to the different sizes of the square and spiral patterns. Also, in both geometric patterns, the reduction of the adhesive glass width induced C54 to create bonds amongst themselves. NIH 3T3 cells adhered inside the squares and the spiral but reducing the adhesive glass width induced NIH 3T3 to adhere to immobilized Hyal. This fact is explained by the interactions between the cells and the immobilized Hyal as a consequence of the CD44/Hyal binding. PMID- 19189384 TI - N-acetyl cysteine prevents polymethyl methacrylate bone cement extract-induced cell death and functional suppression of rat primary osteoblasts. AB - This study examines the cytotoxicity of bone cement extract to osteoblasts and the potential detoxification and restoration of osteoblastic function by an antioxidant amino acid, N-acetyl cysteine (NAC). The osteoblastic cells derived from rat femurs were cultured with extract from polymethyl methacrylate (PMMA) based bone cement. The calcein and ethidium homodimer staining of the cells after 24-h incubation showed that 23.0% of the cells were dead in the culture with bone cement extract, while the addition of 5 mM NAC into the culture reduced the percentage to 4.3%. Annexin V and propidium iodide-based flow cytometric analysis also revealed that the apoptotic cells present at 15.8% in the culture with bone cement extract was reduced to 2.4% in the culture cotreated with bone cement extract and NAC. Severely suppressed alkaline phosphatase activity and matrix mineralization in the culture with bone cement extract (reduced to 10% and 5%, respectively, compared with the control culture) were restored to a normal level when treated with 5 mM NAC. The bone cement extract-induced, downregulated expression of osteoblastic genes, such as alkaline phosphatase, collagen I, and osteocalcin, was also restored to the baseline level by cotreatment with NAC. The data indicated that the addition of NAC into acrylic bone cement extract remarkably ameliorated the cytotoxicity to osteoblasts and restored their phenotype and function to a biologically significant degree, suggesting the potential usefulness of NAC in developing more biocompatible acrylic bone cement. PMID- 19189385 TI - Exogenous crosslinking recovers the functional integrity of intervertebral disc secondary to a stab injury. AB - Exogenous crosslinking was proved to improve the fatigue resistance of anulus fibrosus and the stability of motion segment. The effect of crosslinking on the recovery of stab-injured discs, however, was less studied. The purpose of this study is to find if the exogenous crosslinking can increase the mechanical function of injured discs. Fresh healthy porcine discs (T2/T9) from 6-month-old swine were obtained immediately following death. Anular puncture using 16 and 18 G spinal needle were used to create medium and large disc stab injury models. Three treatments were designed for each injury model. The first one is the injured discs without treatment. The second one is the injured discs soaked with phosphate buffered solution for 2 days. The third one is the injured discs soaked with 0.33% genipin solution for 2 days. The disc integrity was evaluated using quantitative discomanometry (QD) apparatus. Four QD parameters, that is, the leakage pressure and volume, and the saturate pressure and volume, were analyzed to find the efficacy of treatment. We found that soaking of genipin solution recovered the disc leakage pressure from 1.3 to 1.8 MPa in 16 G-injury-model and from 2.3 to 3.2 MPa in 18 G-injury-model, and recovered the saturate pressure from 1.6 to 2.0 MPa in 16 G-injury-model and from 2.7 to 3.7 MPa in 18 G-injury model. The improvement of disc integrity by soaking with genipin solution indicate that the exogenous crosslinking may help the biomechanical performance of an injured disc. PMID- 19189386 TI - Surface modification of PCL-TCP scaffolds improve interfacial mechanical interlock and enhance early bone formation: an in vitro and in vivo characterization. AB - Pretreatment of polycaprolactone-20% tricalcium phosphate (PCL-TCP) scaffolds under alkaline conditions can be utilized to alter surface characteristics for enhanced early bone formation. PCL-TCP scaffolds were treated with sodium hydroxide (NaOH) at various time intervals (group A: untreated, group B: 3M NaOH for 48 h, and group C: 3M NaOH for 96 h). In vitro results showed a greater degree of physical changes in the NaOH-treated scaffolds (B and C) than the untreated group (A). Clearly, the NaOH-treated scaffolds showed an increased surface roughness than the untreated ones. A significantly large number of "channel-like" pits and greater average pit sizes were detected in groups B (14.51 +/- 10.9 microm) and C (20.27 +/- 14.3 microm); and absent in group A. In addition, treated scaffolds had a significant reduction of the water contact angle (40.9-58.2%). Favorably, the pore dimensions and scaffold rod thickness remained unchanged throughout the experiment. When implanted in the calvaria of rabbits, NaOH-treated scaffolds reported greater early matrix deposition and bone formation from scanning electron images and Micro-computed tomography analyses. In conclusion, pretreatment of PCL-TCP scaffolds with NaOH increases the wettability and surface area for initial matrix deposition and early bone ingrowth. PMID- 19189387 TI - Glycolic acid modulates the mechanical property and degradation of poly(glycerol, sebacate, glycolic acid). AB - The development of biodegradable materials with controllable degradation properties is beneficial for a variety of applications. Poly(glycerol-sebacate) (PGS) is a promising candidate of biomaterials; so we synthesize a series of poly(glycerol, sebacate, glycolic acid) (PGSG) with 1:2:0, 1:2:0.2, 1:2:0.4, 1:2:0.6, 1:2:1 mole ratio of glycerol, sebacate, and glycolic acid to elucidate the relation of doped glycolic acid to the degradation rate and mechanical properties. The microstructures of the polymers with different doping of glycolic acid were dissimilar. PGSG with glycolic acid in the ratio of 0.2 displayed an integral degree of ordering, different to those with glycolic acid in the ratio of 0, 0.4, 0.6, and 1, which showed mild phase separation structure. The number, DeltaH(m), and temperature of the PGSG melting peaks tended to decrease with the increasing ratio of doped glycolic acid. In vitro and in vivo degradation tests showed that the degradation rate of PGSG with glycolic acid in the ratio of 0.2 was slowest, but in the ratio range of 0, 0.4, and 0.6, the degradation rate increased with the increase of glycolic acid. All PGSG samples displayed good tissue response and anticoagulant effects. Our data suggest that doping glycolic acid can modulate the microstructure and degree of crosslinking of PGS, thereby control the degradation rate of PGS. PMID- 19189388 TI - Fibronectin activity on substrates with controlled --OH density. AB - Adhesion of human fibroblast to a family of fibronectin (FN) coated model substrates consisting of copolymers of ethyl acrylate and hydroxyl ethylacrylate in different ratios to obtain a controlled surface density of --OH groups was investigated. Cell adhesion and spreading surprisingly decreased as the fraction of --OH groups on the surface increased. AFM studies of FN conformation revealed formation of a protein network on the more hydrophobic surfaces. The density of this network diminished as the fraction of --OH groups in the sample increased, up to a maximal --OH concentration at which, instead of the network, only FN aggregates were observed. The kinetics of network development was followed at different adsorption times. Immunofluorescence for vinculin revealed the formation of well-developed focal adhesion complexes on the more hydrophobic surface (similar to the control glass), which became less defined as the fraction of --OH groups increased. Thus, the efficiency of cell adhesion is enhanced by the formation of FN networks on the substrate, directly revealing the importance of the adsorbed protein conformation for cell adhesion. However, cell-dependent reorganization of substrate-associated FN, which usually takes place on more hydrophilic substrates (as do at the control glass slides), was not observed in this system, suggesting the increased strength of protein-to-substrate interaction. Instead, the late FN matrix formation--after 3 days of culture--was again better pronounced on the more hydrophobic substrates and decreased as the fraction of --OH groups increase, which is in a good agreement with the results for overall cell morphology and focal adhesion formation. PMID- 19189389 TI - Characterization of the spatial immobilization manner of poly(ethylene glycol) to a titanium surface with immersion and electrodeposition and its effects on platelet adhesion. AB - Both terminals of a poly(ethylene glycol) (PEG) molecule were terminated with amines. The PEG was electrodeposited onto titanium (Ti) to give it the biofunctions such as blood compatibility. The effects of the amine of PEG terminals and the pH at PEG solution on the adsorption amount of PEG molecules and the thickness of PEG layer immobilized on the Ti surface were evaluated using quartz crystal microbalance technique and X-ray photoelectron spectroscopy. The interfacial reactivity between PEG terminals and the Ti surface was the largest at pH 11, according to the interaction between the charge of terminal amines of PEG and the point of zero charge of Ti oxide. The orientations of PEG molecules immobilized on the Ti surface with immersion or electrodeposition at pH 11 were determined by Fourier transform infrared reflection absorption spectroscopy. Consequently, the terminal amines of PEG were oriented perpendicularly to the surface in electrodeposition rather than in immersion. The charged PEG randomly immobilized on the Ti surface with immersion led to platelet aggregation, whereas U-shaped PEG molecule immobilized with electrodeposition inhibited platelet adhesion and aggregation. The immobilization manners of PEG on the Ti surface were strongly associated with a biofunction such as platelet adhesion. PMID- 19189390 TI - Relating cell proliferation to in vivo bone formation in porous Ca/P scaffolds. AB - Most current methods for cell monitoring on 3D porous scaffolds involve end-stage investigation of scaffolds. Repeated measurements on scaffolds, without disturbing cell vitality and proliferation, are needed to relate in vitro to in vivo data. Alamar Blue was used for this purpose. Two different Ca/P scaffolds were studied, using rat BMSCs with three different seeding densities [2.5 x 10(4) (SD1), 2.5 x 10(5) (SD2), 2.5 x 10(6) (SD3) cells]. Alamar Blue readings were done on days 1, 3, 5 and 7. After 7 days all 96 scaffolds (n = 16) were implanted in 16 mice for 4 weeks. Bone histomorphometry was performed. For both scaffolds, seeding efficiencies were highest with SD1 and SD2, cell proliferation was optimal in SD1, whereas SD3 resulted in an initial drop in vital cell number in the first 3 days. In vivo, upscaling from SD1 to SD2 lead to significantly more bone contact% in both scaffolds. Alamar Blue was shown to be a valuable tool in relating in vitro to in vivo data. Cell proliferation may differ depending on seeding density and scaffold type used. Seeding more cells may not necessarily result in more in vivo bone contact%. PMID- 19189391 TI - The pore size of polycaprolactone scaffolds has limited influence on bone regeneration in an in vivo model. AB - Bone tissue engineering scaffolds should be designed to optimize mass transport, cell migration, and mechanical integrity to facilitate and enhance new bone growth. Although many scaffold parameters could be modified to fulfill these requirements, pore size is an important scaffold characteristic that can be rigorously controlled with indirect solid freeform fabrication. We explored the effect of pore size on bone regeneration and scaffold mechanical properties using polycaprolactone (PCL) scaffolds designed with interconnected, cylindrical orthogonal pores. Three scaffold designs with unique microarchitectures were fabricated, having pore sizes of 350, 550, or 800 microm. Bone morphogenetic protein-7 transduced human gingival fibroblasts were suspended in fibrin gel, seeded into scaffolds, and implanted subcutaneously in immuno-compromised mice for 4 or 8 weeks. We found that (1) modulus and peak stress of the scaffold/bone constructs depended on pore size and porosity at 4 weeks but not at 8 weeks, (2) bone growth inside pores depended on pore size at 4 weeks but not at 8 weeks, and (3) the length of implantation time had a limited effect on scaffold/bone construct properties. In conclusion, pore sizes between 350 and 800 microm play a limited role in bone regeneration in this tissue engineering model. Therefore, it may be advantageous to explore the effects of other scaffold structural properties, such as pore shape, pore interconnectivity, or scaffold permeability, on bone regeneration when designing PCL scaffolds for bone tissue engineering. PMID- 19189392 TI - Fibronectin-based isolation of valve interstitial cell subpopulations: relevance to valve disease. AB - Myxomatous mitral valves (MVs) contain elevated proportions of unique cell populations such as myofibroblasts. Without a reliable technique to isolate such cell populations, however, it has been difficult to study the role of these cells. The goal of this study was to use fibronectin (FN) to isolate distinct cell subpopulations from normal porcine MVs. Cells from porcine posterior MV leaflets were separated based on time-dependent adhesion to either tissue culture plastic (TCP) flasks or FN-coated flasks. The resultant "FAST" and "SLOW" adhering subpopulations from each technique were phenotyped using flow cytometry and immunocytochemistry to detect expression of myofibroblast markers, enzymes for collagen synthesis, and MAP kinases. Compared with FN SLOW, FN FAST showed significantly higher expression of prolyl 4-hydroxylase, heat shock protein-47 (HSP47), smooth muscle alpha-actin (SMalphaA), nonmuscle myosin (Smem), extracellular-related signaling kinase (ERK) 1, ERK2, and phosphorylated-ERK. In contrast, TCP FAST showed higher expression of only HSP47, SMalphaA, and Smem compared with TCP SLOW. In conclusion, differential adhesion to FN successfully separated a myofibroblast-like subpopulation from the posterior leaflet of the MV. This subpopulation may be useful in studying myxomatous MV disease, although additional studies remain to verify that this myofibroblast-like population resembles that observed in myxomatous MV disease. PMID- 19189393 TI - Multicomponent T2 relaxation analysis in cartilage. AB - MR techniques are sensitive to the early stages of osteoarthritis, characterized by disruption of collagen and loss of proteoglycan (PG), but are of limited specificity. Here, water compartments in normal and trypsin-degraded bovine nasal cartilage were identified using a nonnegative least squares multiexponential analysis of T(2) relaxation. Three components were detected: T(2,1)=2.3 ms, T(2,2)=25.2 ms, and T(2,3)=96.3 ms, with fractions w(1)=6.2%, w(2)=14.5%, and w(3)=79.3%, respectively. Trypsinization resulted in increased (P<0.01) values of T(2,2)=64.2 ms and T(2,3)=149.4 ms, supporting their assignment to water compartments that are bound and loosely associated with PG, respectively. The T(2) of the rapidly relaxing component was not altered by digestion, supporting assignment to relatively immobile collagen-bound water. Relaxation data were simulated for a range of TE, number of echoes, and SNR to guide selection of acquisition parameters and assess the accuracy and precision of experimental results. Based on this, the expected experimental accuracy of measured T(2)s and associated weights was within 2% and 4% respectively, with precision within 1% and 3%. These results demonstrate the potential of multiexponential T(2) analysis to increase the specificity of MR characterization of cartilage. PMID- 19189394 TI - Conditions for partial nitrification in biofilm reactors and a kinetic explanation. AB - Nitrification is a two-step process in which ammonia is incompletely oxidized by ammonia-oxidizing bacteria or archaea (AOB) to nitrite, which is then further oxidized to nitrate by nitrite-oxidizing bacteria (NOB). Literature reports show that segregation of initially coexisting ammonia and nitrite oxidizing populations co-immobilized in gel cubes and cultured in a set-up with three reactors in series (without recirculation) is attained. In those studies NOB were present and nitrite was oxidized mainly in the last reactor. We developed a mathematical model for immobilized biomass that allows for one-dimensional gradients of metabolites and changes of porosity within the gel due to growth. The model reproduced the experimentally observed compartmentalization under the conditions used by Noto et al. (Noto et al., 1998. Water Res 32(3): 769- 773), using standard kinetic parameters of nitrifying bacteria including free ammonia inhibition of AOB and NOB. The model predicted compartmentalization when the ammonium load was sufficiently high and liquid phase mixing sufficiently limited (close to plug-flow). Modeling results demonstrated that inhibition of NOB by free ammonia did not substantially contribute to the compartmentalization in biofilm reactors. Additional simulations identified the higher oxygen affinity of AOB as the key parameter leading to compartmentalization (i.e., partial nitrification) in artificial and natural biofilms since they enable the formation of oxygen gradients. As a result, a tendency for compartmentalization was found even at equal competitiveness. PMID- 19189395 TI - Simultaneous analysis of physiological and electrical output changes in an operating microbial fuel cell with Shewanella oneidensis. AB - Changes in metabolism and cellular physiology of facultative anaerobes during oxygen exposure can be substantial, but little is known about how these changes connect with electrical current output from an operating microbial fuel cell (MFC). A high-throughput voltage based screening assay (VBSA) was used to correlate current output from a MFC containing Shewanella oneidensis MR-1 to carbon source (glucose or lactate) utilization, culture conditions, and biofilm coverage over 250 h. Lactate induced an immediate current response from S. oneidensis MR-1, with both air-exposed and anaerobic anodes throughout the duration of the experiments. Glucose was initially utilized for current output by MR-1 when cultured and maintained in the presence of air. However, after repeated additions of glucose, the current output from the MFC decreased substantially while viable planktonic cell counts and biofilm coverage remained constant suggesting that extracellular electron transfer pathways were being inhibited. Shewanella maintained under an anaerobic atmosphere did not utilize glucose consistent with literature precedents. Operation of the VBSA permitted data collection from nine simultaneous S. oneidensis MR-1 MFC experiments in which each experiment was able to demonstrate organic carbon source utilization and oxygen dependent biofilm formation on a carbon electrode. These data provide the first direct evidence of complex cellular responses to electron donor and oxygen tension by Shewanella in an operating MFC at select time points. PMID- 19189396 TI - Alignment of skeletal muscle myoblasts and myotubes using linear micropatterned surfaces ground with abrasives. AB - Alignment of cells plays a significant key role in skeletal muscle tissue engineering because skeletal muscle tissue in vivo has a highly organized structure consisting of long parallel multinucleated myotubes formed through differentiation and fusion of myoblasts. In the present study, we developed an easy, simple, and low-cost method for aligning skeletal muscle cells by using surfaces with linear microscale features fabricated by grinding. Iron blocks were ground in one direction with three kinds of abrasives (9 microm diamond suspension, #400 sandpaper, and #150 sandpaper) and then used as molds to make micropatterned polydimethylsiloxane (PDMS) substrates (type I, type II, and type III). Observation of the surface topography revealed that the PDMS substrates exhibited different degree of mean roughness (Ra), 0.03 microm for type I, 0.16 microm for type II, and 0.56 microm for type III, respectively. Murine skeletal muscle cell line C2C12 myoblasts were cultured and differentiated on the patterned PDMS substrates, and it was examined whether the alignment of C2C12 myoblasts and myotubes was possible. Although the cell growth and differentiation on the three types of patterned substrates were similar to those on the flat PDMS substrate as a control, the alignment of both C2C12 myoblasts and myotubes was obviously observed on types II and III, but not on type I or the control substrate. These results indicate that surfaces ground with abrasives will be useful for fabricating aligned skeletal muscle tissues. PMID- 19189397 TI - The effect of starch material, encapsulated protein and production conditions on the protein release from starch microspheres. AB - The present study describes the preparation of 11 batches of starch microspheres for drug delivery. Parameters such as the type of starch material, the type of protein, and the incubation time of the process were varied, and the obtained microspheres differed in yield, encapsulation efficiency and physical properties. The crystalline/ordered structure (obtained through X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC)), the microscopic appearance and the surface morphology (viewed with scanning electron microscopy (SEM)) were found to differ between the batches depending on the starch type, encapsulated protein and incubation conditions that were employed. Freeze-drying was found to have a destructive effect on the ordered structure of the starch and this effect varied with regard to preparation conditions. Drug release experiments demonstrated that the release from the starch matrix depended on the type of protein as well as on the incubation time during the manufacturing at temperatures of 6 degrees C and 37 degrees C. The enzymatic degradation of starch was slightly different between the materials depending on the crystalline/ordered structure that had formed during the preparation. PMID- 19189398 TI - Impact of polymeric nanoparticles on oral pharmacokinetics: a dose-dependent case study with estradiol. AB - The present study reports the significance of polymeric nanoparticles in oral drug delivery from pharmacokinetic perspective. Impact of different dose polymeric nanoparticulate formulations on various pharmacokinetic parameters has been reported. All nano-formulations were found to have significant effect on important pharmacokinetic parameters like C(max), T(max), AUC, absolute bioavailability, absorption rate constant (K(a)), elimination rate constant (K(el)), elimination half life (t(1/2)), mean residence time (MRT) and mean absorption time (MAT). Further, drug in polymeric nanoparticles exhibited the flip-flop pharmacokinetics suggesting its slow and sustained release from the polymeric matrix. Together, polymeric nanoparticles hold promise for the oral delivery of molecules having pharmacokinetic hurdles in their delivery approach. PMID- 19189399 TI - Nonlinear quantitative structure-property relationship modeling of skin permeation coefficient. AB - The permeation coefficient characterizes the ability of a chemical to penetrate the dermis, and the current study describes our efforts to develop structure based models for the permeation coefficient. Specifically, we have integrated nonlinear, quantitative structure-property relationship (QSPR) models, genetic algorithms (GAs), and neural networks to develop a reliable model. Case studies were conducted to investigate the effects of structural attributes on permeation using a carefully characterized database. Upon careful evaluation, a permeation coefficient data set consisting of 333 data points for 258 molecules was identified, and these data were added to our extensive thermophysical database. Of these data, permeation values for 160 molecular structures were deemed suitable for our modeling efforts. We employed established descriptors and constructed new descriptors to aid the development of a reliable QSPR model for the permeation coefficient. Overall, our new nonlinear QSPR model had an absolute average percentage deviation, root-mean-square error, and correlation coefficient of 8.0%, 0.34, and 0.93, respectively. Cause-and-effect analysis of the structural descriptors obtained in this study indicates that that three size/shape and two polarity descriptors accounted for approximately 70% of the permeation information conveyed by the descriptors. PMID- 19189400 TI - Topical oral cavity pharmacokinetic modeling of a stannous fluoride dentifrice: an unusual two compartment model. AB - A pharmacokinetic model was developed describing the pharmacokinetics of stannous fluoride in human subjects after oral topical application of a stannous fluoride dentifrice. Twenty subjects participated in an investigation of an experimental dentifrice. Subjects rinsed their mouths with the experimental dentifrice slurry. Saliva and plaque samples were obtained from the subjects at various times up to 6 h after administration. Samples were analyzed for total tin content, used as an analytical marker for the active stannous fluoride ingredient, using a graphite furnace atomic absorption spectrometer. The modeling indicates that there is an obvious kinetic relationship between saliva and plaque compartments and that stannous fluoride is very well retained in and slowly released from plaque (and oral surfaces) into saliva. Additionally, both compartments are simultaneously loaded during administration unlike typical systemic drug behavior, and the elimination rate "constant" from the central compartment (saliva) changes due to changes in salivary flow. Stannous fluoride is cleared from saliva rapidly but very well retained in gingival plaque. The model with simultaneous loading of plaque and saliva describes these observations and may account for the prolonged antiplaque and antigingivitis benefits of stannous fluoride. PMID- 19189401 TI - Potent antitumor effect of SN-38-incorporating polymeric micelle, NK012, against malignant glioma. AB - Recent published reports on clinical trials of CPT-11 indicate the effectiveness of this compound, a prodrug of SN-38, against malignant glioma in combination with anti-vascular endothelial growth factor antibody. Here, we determined if NK012, and SN-38 incorporating micelle, can be an appropriate formulation for glioblastoma treatment compared with CPT-11. In vitro cytotoxicity was evaluated against several glioma lines with NK012, CPT-11, SN-38, ACNU, CDDP and etoposide. For the in vivo test, a human glioma line (U87MG) transfected with the luciferase gene was inoculated into nude mice brain for pharmacokinetic analysis by fluorescence microscopy and high-performance liquid chromatography after intravenous injection of NK012 and CPT-11. In vivo antitumor activity of NK012 and CPT-11 was evaluated by bioluminescence image and Kaplan-Meier analyses. The growth-inhibitory effects of NK012 were 34- to 444-fold more potent than those of CPT-11. Markedly enhanced and prolonged distribution of free SN-38 in the xenografts was observed after NK012 injection compared with CPT-11. NK012 showed significantly potent antitumor activity against an orthotopic glioblastoma multiforme xenograft and significantly longer survival rate than CPT-11 (p = 0.0014). This implies that NK012 can pass through the blood brain tumor barrier effectively. NK012, which combines enhanced distribution with prolonged sustained release, may be ideal for glioma treatment. Currently, a phase I study of NK012 is almost complete in Japan and the US. The present translational study warrants the clinical phase II study of NK012 in patients with malignant glioma. PMID- 19189402 TI - Human papillomavirus type distribution in anal cancer and anal intraepithelial lesions. AB - A systematic review was conducted of HPV type distribution in anal cancer and anal high-grade and low-grade squamous intraepithelial lesions (HSIL and LSIL). A Medline search of studies using PCR or hybrid capture for HPV DNA detection was completed. A total of 1,824 cases were included: 992 invasive anal cancers, 472 HSIL cases and 360 LSIL cases. Crude HPV prevalence in anal cancer, HSIL, and LSIL was 71, 91 and 88%, respectively. HPV16/18 prevalence was 72% in invasive anal cancer, 69% in HSIL and 27% in LSIL. The HPV 16 and/or 18 prevalence in invasive anal cancer cases was similar to that reported in invasive cervical cancer. If ongoing clinical trials show efficacy in preventing anal HPV infection and associated anal lesions, prophylactic HPV vaccines may play an important role for the primary prevention of these cancers in both genders. PMID- 19189403 TI - Circulating prediagnostic interleukin-6 and C-reactive protein and prostate cancer incidence and mortality. AB - Interleukin-6 (IL-6) and C-reactive protein (CRP) are elevated in prostate cancer patients, but the role of prediagnostic levels of these inflammatory mediators on prostate cancer outcomes is unclear. We undertook a large, prospective case control study to evaluate the relation between prediagnostic levels of IL-6 and CRP and prostate cancer incidence and mortality. We also investigated the role of the IL-6 (-174 G/C) polymorphism in relation to circulating levels of IL-6 and CRP, as well as cancer risk and mortality. We used unconditional logistic regression that adjusted for matching factors to analyze prostate cancer risk. For analyses of prostate cancer mortality, we conducted survival analyses in cases. Because of the strong link between inflammatory markers and body mass index (BMI), we assessed interactions between BMI and plasma levels on prostate cancer outcomes. Neither IL-6 nor CRP plasma levels varied significantly by IL-6 genotype. Genotype was not associated with prostate cancer risk or survival. Though neither IL-6 nor CRP was associated with prostate cancer incidence overall, we observed a statistically significant interaction between IL-6 and BMI on prostate cancer incidence (p(interaction) < 0.01). Increasing IL-6 levels were positively associated with risk in healthy weight men, but inversely associated with risk in overweight men. Further, prediagnostic IL-6 was associated with time to prostate cancer progression/death among healthy weight prostate cancer cases (p(trend) = 0.02). Adjusted hazard ratios were 1.73 (95% CI: 0.86, 3.51) comparing the highest to lowest IL-6 level. Our study suggests that IL-6 may potentially be involved in the development or progression of prostate cancer. PMID- 19189405 TI - Caffeine, artificial sweetener, and fluid intake in anorexia nervosa. AB - OBJECTIVE: This article provides an analysis of the use of artificial sweeteners, caffeine, and excess fluids in patients diagnosed with anorexia nervosa (AN). METHOD: Seventy participants with AN were recruited to participate in an ecologic momentary assessment study which included nutritional analysis using the Nutrition Data Systems for Research, a computer based dietary recall system. RESULTS: When subtypes were compared, participants with AN-restricting subtype (AN-R) and participants with AN-Binge-Purge (AN-B/P) did not differ in quantity of aspartame, caffeine, or water consumed. Daily water consumption was related to daily vomiting frequency in AN-B/P but not to daily exercise frequency in either participants with AN-R or AN-B/P. DISCUSSION: Caffeine, water, and aspartame consumption can be variable in patients with AN and the consumption of these substances seems to be only modestly related to purging behavior. PMID- 19189406 TI - Second-degree atrioventricular block (Mobitz Type I) in an adolescent with anorexia nervosa: intrinsic or acquired conduction abnormality. AB - Anorexia nervosa (AN) can cause both functional and structural cardiac complications, including a variety of different conduction abnormalities. This is the first case report of symptomatic diurnal second-degree atrioventricular (AV) block (Mobitz Type I) in an adolescent with AN. We present a 12-year-old girl with AN, restrictor sub-type who reported cardiac symptoms during weight gain, at the time of the initial diagnosis of AV block. Second-degree AV block (Mobitz Type I) is discussed as a possible complication of the AN, as well as being an intrinsic conduction system disease. PMID- 19189407 TI - Family functioning and maternal distress in adolescent girls with anorexia nervosa. AB - OBJECTIVE: To determine the nature of family distress in families of girls with anorexia nervosa (AN), this study compared aspects of family functioning in adolescent girls with AN to girls with a chronic illness and girls without a condition. METHOD: Participants consisted of 25 adolescent girls with a primary DSM-IV diagnosis of AN, 20 girls with an ICD-10 diagnosis of insulin dependent diabetes mellitus (IDDM), and 20 girls from the community. Mothers and daughters completed questionnaires of family functioning and psychological symptoms. RESULTS: When compared with mothers of daughters with IDDM, families of girls with AN experienced greater family conflict, reduced parental alliance, and increased feelings of depression. However, once the emotional impact of the illness on the mothers was statistically controlled, group differences were no longer significant. DISCUSSION: Family distress and dysfunction may reflect an accommodation process that occurs in families living with a child with AN. PMID- 19189408 TI - Treating childhood obesity: family background variables and the child's success in a weight-control intervention. AB - OBJECTIVE: To analyze whether caregiver and family characteristics predict success in a family-based lifestyle intervention program for children and adolescents. METHOD: Participants were 111 overweight and obese children (7-15 years) who attended a family-based weight-reduction program. Body mass index (BMI) and BMI standard deviation scores (BMI-SDS) of index child, and BMI of family members, family adversity characteristics, depression, and attachment attitudes of the primary caregiver were assessed. RESULTS: Risk of nonresponse ( 0.05, Student's t-test) were observed in growth between studied children. Multiple regression analyses showed age, sex, mother's height, birth order, birth weight, family income, zone of residence, diet, and febrile episodes had an effect on growth. Neither the migration variable used above nor any other definition of migrant had a significant effect on growth. The lack of differences in growth between immigrant and native children is probably due to similarity in socioeconomic conditions of their families. PMID- 19189417 TI - Changes in serum immunity during pregnancy. AB - Pregnancy requires a host of localized immune factors that allow the mother to tolerate the fetus. Changes in the mother's serum immunity during pregnancy are less well-known. To clarify these changes, 1,351 women from the NHANES 1999-2000 were analyzed with complex survey regression to test the effect of pregnancy on adaptive and innate immune markers. Adjusting for age and BMI, pregnant women had higher C-reactive protein levels and white blood cell counts and lower measles antibody titer and lymphocyte counts than nonpregnant women. This dual pattern of immunological changes supports the hypothesis that mothers will reduce the ability of the adaptive immune system to respond to infection while increasing the activity of innate immunity during pregnancy, maintaining immune function homeostasis. The function of these homeostatic immune responses is unknown. PMID- 19189418 TI - Long sleep duration and childhood overweight/obesity and body fat. AB - To assess the association between short sleep duration and overweight/obesity and body fat (BF) and to identify correlates of short sleep duration in a sample of Portuguese children. A cross-sectional study of children 7-9 years (n = 4511) was performed between October 2002 and June 2003. Weight, height, and skinfolds were measured, and parents filled out a questionnaire about family characteristics as well as sleep duration. The prevalence of overweight/obesity and BF (%) both decreased by long sleep duration. After adjusted for confounders, the odds ratio (OR) for overweight/obesity and sleep duration were as follows: reference >11 h/d; 10-11 h/d, OR: 1.3; confidence interval (CI):1.26, 1.33; 9-10 h/d, OR: 1.16; CI: 1.13, 1.19; and <9 h, OR: 3.22; CI: 3.11, 3.32. Children whose parents had a low educational level slept less time during each night than children whose parents had a higher educational level; children who spent more time watching television slept less time than those who watched less television, and those children engaged in physical activity slept more time each night than sedentary children. Our results showed an inverse relationship between long sleep duration and overweight/obesity prevalence as well as with body fat, and these findings are important because sleep duration is a potentially modifiable risk factor that could be important to consider in the prevention and treatment of childhood obesity. PMID- 19189419 TI - Thermosensitive PEG-PCL-PEG hydrogel controlled drug delivery system: sol-gel-sol transition and in vitro drug release study. AB - In this article, biodegradable and low molecular weight poly(ethylene glycol) poly(epsilon-caprolactone)-poly(ethylene glycol) (PEG-PCL-PEG, PECE) triblock copolymers were successfully synthesized. Aqueous solution of the obtained PECE copolymers underwent sol-gel-sol transition as temperature increased which was flowing sol at room temperature and then turned into nonflowing gel at body temperature. Sol-gel-sol phase transition behaviors of aqueous PECE solutions were studied using rheometry and test tube-inverting method, which were affected by many factors, including the heating/cooling procedure and different additives in copolymers aqueous solution. In vitro drug release behavior was studied using bovine serum albumin (BSA) and Vitamin B(12) (VB(12)) as model drugs, and the PECE hydrogel could protect BSA from acidic degradation for 1 week at least. Therefore, PECE hydrogel is believed to be promising for injectable in situ gel forming controlled drug delivery system due to their great thermosensitivity and biodegradability. PMID- 19189420 TI - Simultaneously manufactured nano-in-micro (SIMANIM) particles for dry-powder modified-release delivery of antibodies. AB - Simultaneously Manufactured Nano-In-Micro (SIMANIM) particles for the pulmonary delivery of antibodies have been prepared by the spray-drying of a double emulsion containing human IgG (as a model antibody), lactose, poly(lactide-co glycolide) (PLGA) and dipalmitoylphosphatidylcholine (DPPC). The one-step drying process involved producing microparticles of a diameter suitable for inhalation that upon contact with aqueous media, partially dissolved to form nanoparticles, approximately 10-fold smaller than their original diameter. Continuous release of the model antibody was observed for 35 days in pH 2.5 release media, and released antibody was shown to be stable and active by gel electrophoresis, field-flow fractionation and enzyme linked immunosorbent assay. Adding 1% L-leucine to the emulsion formulation, and blending 'SIMANIM' particles with 1% magnesium stearate, achieved a fine particle fraction of approximately 60%, when aerosolised from a simple, capsule-based, dry powder inhaler device. 'SIMANIM' particles could be beneficial for the delivery of antibodies targeted against inhaled pathogens or other extracellular antigens, as well as having potential applications in the delivery of a wide range of other biopharmaceuticals and certain small-molecule drugs. PMID- 19189421 TI - Celebrating 10 years of hESC lines: an interview with Ariff Bongso. PMID- 19189422 TI - Celebrating 10 years of hESC lines: an interview with Martin Pera. PMID- 19189424 TI - Statistically significant RCT results are not always possible: should case-series studies inform practice? PMID- 19189423 TI - The Sorghum bicolor genome and the diversification of grasses. AB - Sorghum, an African grass related to sugar cane and maize, is grown for food, feed, fibre and fuel. We present an initial analysis of the approximately 730 megabase Sorghum bicolor (L.) Moench genome, placing approximately 98% of genes in their chromosomal context using whole-genome shotgun sequence validated by genetic, physical and syntenic information. Genetic recombination is largely confined to about one-third of the sorghum genome with gene order and density similar to those of rice. Retrotransposon accumulation in recombinationally recalcitrant heterochromatin explains the approximately 75% larger genome size of sorghum compared with rice. Although gene and repetitive DNA distributions have been preserved since palaeopolyploidization approximately 70 million years ago, most duplicated gene sets lost one member before the sorghum-rice divergence. Concerted evolution makes one duplicated chromosomal segment appear to be only a few million years old. About 24% of genes are grass-specific and 7% are sorghum specific. Recent gene and microRNA duplications may contribute to sorghum's drought tolerance. PMID- 19189425 TI - Financing R&D for neglected diseases. PMID- 19189426 TI - [Cholera outbreak in Zimbabwe]. PMID- 19189428 TI - New hopes for US health pinned on Daschle's nomination. PMID- 19189429 TI - UK doctors hail research excellence results. PMID- 19189427 TI - Diffusion-weighted magnetic resonance imaging of head and neck squamous cell carcinomas. AB - OBJECTIVE: To evaluate whether diffusion-weighted imaging (DWI) is a reliable technique to quantify microstructural differences between head and neck squamous cell carcinomas (SCC) and tumour-free soft tissue. MATERIALS AND METHODS: DWI was obtained from 20 patients with histologically proven, untreated head and neck SCC. DWI was acquired using a diffusion-weighted, navigated echo-planar imaging sequence with a maximum b-value of 800 s/mm2. For an objective assessment of image quality, the signal-to-noise ratio (SNR) was calculated. Microstructural differences between vital tumour tissue and tumour-free soft tissue were quantified by calculating the apparent-diffusion-coefficients (ADC) on a pixel by pixel method. RESULTS: Echo-planar DWI provided good image quality in all patients (mean SNR 18.4). The mean ADC of SCC, (0.64+/-0.28 x 10(-3) mm2/s), was significantly (P<0.0001) lower than that of the tumour-free soft tissue, (2.51+/ 0.82 x 10(-3) mm2/s). CONCLUSION: DWI is a reliable diagnostic tool to quantify the microstructural differences between vital tumour tissue and tumour-free soft tissue in patients with head and neck SCC. PMID- 19189430 TI - Plague and theatre in ancient Athens. PMID- 19189431 TI - Erratum to "Sonographic findings of the median nerve and prevalence of carpal tunnel syndrome in patients with Parkinson's disease" [Eur. J. Radiol. 67 (3) (2008) 546-550]. AB - PURPOSE: Parkinson's disease (PD) is a chronic progressive disorder which is characterized by rest tremor, akinesia or bradykinesia and rigidity. Carpal tunnel syndrome (CTS) is caused by compression of median nerve and can occur as a result of repetitive trauma. The aim of this study was to estimate the prevalence of CTS in PD and evaluate the median nerve sonographically. MATERIALS AND METHODS: Fifty-three wrist of 29 patients with PD were included in the study according to Hoehn and Yahr (H&Y) clinical stage and divided into two groups. The first group consisted of 29 wrists of patients with mild PD (H&Y stage I-II). The second group consisted of 24 wrists of patients with severe PD (H&Y stage III IV). Thirty-six wrists of 20 age-matched patients were used as control group. Both of the patients with PD and control group underwent sonography and electromyography (EMG). Axial sonograms of the median nerve were obtained at the level of distal radioulnar joint (level 1) and at the level of pisiform bone in the carpal tunnel (level 2). At each level, the cross-sectional area of the median nerve and flattening ratio were calculated. RESULTS: There was no significant difference for all parameters, except one parameter, between the patients with PD and control group, and also among mild and severe groups of PD and control group (p > 0.05). Interestingly, amplitude of median nerve in the second finger was significantly lower in PD patients than control group within normal limits (p=0.010). Of all wrists of PD patients, 13 (24.4%) have been diagnosed as CTS and 7 (19.4%) control wrists had CTS. Median nerve cross sectional area of CTS patients were more than 10 mm2 in 6 (46%) wrists of PD patients but in only 1 (14%) control wrist at each level. Although it was not statistically significant, there was higher cross-sectional area at each level in patients with severe PD (level 1: 10.43+/-2.30 mm2, level 2: 10.35+/-3.19 mm2) than patients with mild PD (level 1: 9.93+/-2.61 mm2, level 2: 9.51+/-2.83 mm2) and control group (level 1: 9.69+/-3.19 mm2, level 2: 9.07+/-3.61 mm2). CONCLUSION: PD may pose a risk for the development of CTS due to the repetitive movement of tremor. Although sonography is not an ideal method of diagnosis for CTS, it may take our attention for indicating CTS in patients with PD especially in severe ones. PMID- 19189432 TI - Erratum to "Carpal tunnel syndrome in Parkinson's disease" [Eur. J. Radiol. 67 (3) (2008) 550]. PMID- 19189433 TI - Announcements from your Editorial Team. PMID- 19189434 TI - Words of wisdom. Re: hypothalamic-pituitary-testicular axis disruptions in older men are differentially linked to age and modifiable risk factors: the European Male Aging Study. PMID- 19189435 TI - Words of wisdom. Re: phase I clinical trial of a selective inhibitor of CYP17, abiraterone acetate, confirms that castration- resistant prostate cancer commonly remains hormone driven. PMID- 19189436 TI - Words of wisdom. Re: utilization and outcomes of minimally invasive radical prostatectomy. PMID- 19189437 TI - Words of wisdom. Re: long-term outcome following three-dimensional conformal/intensity-modulated external- beam radiotherapy for clinical stage T3 prostate cancer. PMID- 19189438 TI - Words of wisdom. Re: enucleation of renal cell carcinoma with ablation of the tumour base. PMID- 19189439 TI - Words of wisdom. Re: predictors of intravesical therapy for nonmuscle invasive bladder cancer: results from the Surveillance, Epidemiology and End Results Programme (SEER) 2003 Patterns of Care Project. PMID- 19189440 TI - Expression of cadherin superfamily genes in brain vascular development. AB - Cadherins are Ca2+-dependent cell adhesion molecules that are important in vertebrate nervous system development. We identified seven members of the cadherin superfamily (cadherin-4, cadherin-5, cadherin-6, cadherin-6, cadherin 11, protocadherin-1, and protocadherin-17) and an intracellular binding partner of delta-protocadherins, protein phosphatase 1alpha, as novel markers for developing blood vessels in the ferret brain. Some of the cadherin molecules are restricted to specific brain regions or a subset of blood vessels. The expression levels show a peak during perinatal vascular development. Our results suggest that multiple cadherins, which are also involved in neurogenesis, are regulators of angiogenesis in developing vertebrate brain. PMID- 19189442 TI - Approved: special scoring consideration for critical access hospitals through 2008. PMID- 19189443 TI - 2008 salary survey results. Within the growing role of QI, technology playing a big part. PMID- 19189447 TI - [Mandatory retirement for physicians eliminated: however, the law remains]. PMID- 19189448 TI - [Check up, skin cancer screening, vaccination: prevention concepts firmly integrated in general practice]. PMID- 19189449 TI - [Healthy nutrition for heart and blood vessels: fruit, vegetables and hibiscus tea have protective value, vitamin pills are ineffective]. PMID- 19189450 TI - [Chronic heart failure: sports improve the prognosis]. PMID- 19189451 TI - [Ameliorating strabismus, preventing blepharospasm: botulinum toxin can do more than smooth wrinkles]. PMID- 19189452 TI - [Lipid metabolism disorders in children and the elderly. Therapy at the fringes of the indication range]. PMID- 19189454 TI - [Physical activity and obesity]. PMID- 19189453 TI - [Treatment of hyperlipidemia in the elderly]. PMID- 19189455 TI - [How to interpret pulmonary symptoms in elderly patients]. PMID- 19189456 TI - [Fall prevention: differences between native vitamin D, alfacalcidol and D hormone]. PMID- 19189457 TI - [Endothelial dysfunction in children's chronic glomerulonephritis]. AB - Production of NO, sVCAM-1, Et-1, FV, PAI- 1 was measured in blood sera of children with different forms of chronic glomerulonephritis. Endothelial dysfunction (ED) occurred in the active phase of the disease and persisted during remission regardless of the clinical form of glomerulonephritis. Severity of ED and involvement of its markers depended on morphological type of the disease. Production of endothelin-1 was highest in patients with sclerotic glomerulonephritis and accompanying arterial hypertension. ED caused activation of hemostasis with the prevalence of prothrombotic condition. ED contributed to the progress of glomerulonephritis acting via sclerotization and cell proliferation. Selected markers ED are shown to be of prognostic value in children with chronic glomerulonephritis. PMID- 19189458 TI - [Immunophenotypic characteristics of blood lymphocytes and expression of cytokine receptors in newborn infants]. AB - Formation of the immune system during the neonatal period was studied in 88 infants aged 3 days-1 month (gestation age 28-41 weeks) with the primary diagnosis of cerebral ischemia. The patients were allocated to 4 groups based on chronological and gestational age at the moment of examination and a reference group. A total of 16 umbilical blood samples were available for analysis. Phenotype of blood lymphocyte was identified by flow cytofluorimetry with differentiation between subpopulations of T and B lymphocytes and detection of membrane receptors of IL-2, 8, and 10. T-cell subpupulation was found to be enriched in CD3+CD4+CD8+ lymphocytes. Content of B1 lymphocytes was equally high in full-term and premature infants. The latter showed enhanced expression of IL-2 receptor, CD122, by the end of the first month of life. Expression of receptors of IL-8 (chemotaxis factor) and IL-10 was high in lymphocytes of children of all groups since the onset of the neonatal period. It is concluded that comprehensive study of membrane cytokine receptors is of interest for better understanding mechanisms underlying maturation of the immune system in children of different age groups. PMID- 19189460 TI - [Role of occupational pathology in industrial medicine system]. PMID- 19189459 TI - [Autoimmune mechanisms of modulation of the activity of glutamate receptors in children with epilepsy and craniocerebral injury]. AB - The role of glutamate receptors and their hyperstimulation in the development of autoimmune processes is discussed with reference to brain pathology associated with hypoxia and ischemia. Epilepsy, paroxismal condition, and craniocerebral injury (CCI) in children are shown to be accompanied by a rise in the levels of antibodies against AMPA and NMDA receptors of glutamate and nitric oxide markers (cGMP, nitrates + nitrites). Also enhanced in epilepsy and paroxismal condition are the levels of cGMP and antibodies against AMPA(GluR1) receptors of glutamate. Acute CCI period is characterized by a marked change in the levels of NO metabolites and antibodies to two subtypes of glutamate receptor, AMPA and NMDA. The levels of antibodies to NMDA(NR2A) receptors are significantly different within 1 day after CCI depending on its outcome. Unfavourable outcome of CCI is associated with the lowest level of antibodies and high NO metabolite content. Relationship between the levels of NO and antibodies against glutamate receptors is discussed with the use of experimental data. It is concluded that antibodies to glutamate receptors and receptor hyperstimulation play an important role in pathogenesis of hypoxia. PMID- 19189461 TI - [Study of genetic control of pediocins synthesis]. AB - Plasmid profiles of 2 strains of Pediococcus acidilactici and 3 strains of P. pentosaceus were studied. Synthesis of bacteriocins in 2 strains of P. acidilactici was coded on the plasmid which was confirmed by experiment with elimination of plasmids and subsequent electrophoregram, which demonstrated synchronous disappearance of bacteriocins synthesis characteristic in plasmid lacking variants. In 3 strains of P. pentosaceus plasmids were not found which points to chromosomal localization of genes for bacteriocins production. PMID- 19189462 TI - [Modification of bacterial population structure on anti-lysozyme activity under the influence of hexylresorcinol]. AB - AIM: To investigate the modifying effect of hexylresorcinol chemical analog on anti-lysozyme activity of bacteria. MATERIALS AND METHODS: Influence of hexylresorcinol on anti-lysozyme activity (ALA) of bacteriawas studied on the examples of such strains as Bacillus cereus 279, Klebsiella pneumoniae 278, Escherichia coli 101, which are able to inactivate lysozyme. As a chemical analog of microbial regulators, C6-alkyloxybenzol (C6-AOB) was used, which was introduced in growth medium containing microorganisms, which than were cultivated until entering in the stationary growth phase. Isolation of clones was carried out by seeding the 24-hours broth culture on solid growth medium followed by measurement of ALA with photometric method. RESULTS: Modifying effect of chemical analog of extracellular microbial regulators (C6-AOB) on population structure of bacteria on the sign of anti-lysozyme activity was revealed. Maximal effect was observed at C6-AOB concentrations 1 and 10 mcg/ml. Clonal redistribution on ALA manifested by decrease of proportion of clones with high and intermediate values of ALA and increase of proportion of clones with low ALA or its absence. CONCLUSION: Obtained data are perspective for development of measures for control of persisting pathogenic microorganisms by modification of persistence factors with microbial autoregulators. PMID- 19189463 TI - [Obtaining of outer membrane protein I of Pseudomonas aeruginosa and assessment of its antigenic properties]. AB - Gene of outer membrane protein I (OprI) of Pseudomonas aeruginosa was cloned in Escherichia coli cells. Synthesized protein OprI contained additional sequence of 6 histidines on the N-terminus, which allowed its chromatographic purification in Ni-agarose. Obtained recombinant ptotein specifically reacted with hyperimmune rabbit serum against whole-cell P. aeruginosa and stimulated synthesis of specific antibodies in immunized mice and rabbits. Obtained hyperimmune rabbit sera against recombinant protein OprI had directive antimicrobial activity against P. aeruginosa. PMID- 19189464 TI - [Development of experimental test-system on the basis of immunochip for syphilis serodiagnostics]. AB - AIM: Development of test-system on the basis of immunochip for detection of IgG to Treponema pallidum. MATERIALS AND METHODS: Recombinant T. pallidum antigens Tp47, Tp17, Tp15, TmpA were separately immobilized on activated slides as individual spots. The specific antibodies to T. pallidum in human serum were detected by indirect immunofluorescent assay on the immunochip using antispecies antibodies to human IgG. Fluorescent scanner was used to read results of testing. For each antigen threshold level of fluorescent signal and positivity coefficient were calculated. Assessment of specificity and sensitivity of the immunochip was performed on 400 human serum samples containing or not-containing antibodies to T. pallidum. RESULTS: From 200 positive serum samples 5 were interpreted as inconclusive because positive results on antigen Tp17 only were registered that was comparable with the results of immunoblotting. For each other 195 serum samples, positive result on 2 or more antigens was obtained. Specificity of the immunochip-based test-system was 100%. CONCLUSION: Experimental immunochip-based test-system for differential serologic diagnostics of the syphilis was developed. Its sensitivity and specificity are comparable with that of ELISA-based test system. PMID- 19189465 TI - [Characteristic of resistance to beta-lactam antibiotics of nosocomial strains of Proteus mirabilis]. AB - Antibiotic resistance of bacterium Proteus mirabilis isolated from inpatients was assessed in 30 medical centres from 15 regions of Russia. The most active antibacterial preparations remained carbapenems (imipenem and meropenem). Between 3rd generation cephalosporins, the lowest minimal inhibitory concentration (MIC) was observed in inhibition-protected preparations (ceftazidime/clavulanate: MIC50 < 0.125 mcg/ml, MIC90 128 mcg/ml; cefoperazone/sulbactam: MIC50 2 mcg/ml, MIC90 64 mcg/ml). Detection of class A beta-lactamases genes (TEM, SHV, CTX) using PCR was performed in 44 strains of P. mirabilis. Genes of these enzymes was detected in 11 (25%) bacterial cultures as well as their different combinations--in 32 (72.7%) isolates of P. mirabilis. The most prevalent (56.8%) was combination of TEM and CTX genes, and in 4 nants of resistance to beta-lactams was detected. PMID- 19189466 TI - [Legal and theoretical premises of use of nanotechnology and nanomaterials in diagnostics, prevention and treatment of quarantine infectious diseases]. AB - AIM: Analysis of legal acts designed for progress of nanotechnologies in Russia and in basic works on use of nanotechnologies and nanomaterials for development of methods for diagnostics, prevention and treatment of especially dangerous infections in Russia. MATERIALS AND METHODS: Nanoparticles, nanostructural materials, atomic-force microscopy. RESULTS: In 2007 set of normative documents regulating conduction of works in area of nanotechnology was adopted. Now in research and scientific institutes of Federal Service for Surveillance for Protection of Consumers Rights and Human Welfare, Russian Academy of Medical Sciences and other agencies works are conducted in order to develop methods for diagnostics, prevention and treatment of especially dangerous infectious diseases with applying nanotechnologies and nanomaterials. CONCLUSION: Incorporation of nanotechnologies in health care practice will provide qualitatively new level of struggle with especially dangerous infectious diseases. PMID- 19189467 TI - [Characteristic of sexually-transmitted diseases]. AB - Classification and clinical and epidemiological characteristics of diseases with mainly sexual route of transmission are presented. Their high social and economical significance is showed as well as reasons of high prevalence of these diseases in Russia are demonstrated. Modern ways of prevention and control of sexually-transmitted diseases in different population groups are outlined. PMID- 19189468 TI - Scope and standards for transplant nursing. PMID- 19189469 TI - Short-term cost cutting is not the answer. PMID- 19189470 TI - Mental health is at crossroads as the National Service Framework comes to an end. PMID- 19189471 TI - A rocky road. PMID- 19189472 TI - The highs and lows of rural life. PMID- 19189473 TI - Recovery: the true meaning of recovery. PMID- 19189474 TI - Flying doctor: why I left the UK. PMID- 19189475 TI - Can patient choice ever be a reality in mental health? PMID- 19189476 TI - 'Tis the season to be jolly. PMID- 19189477 TI - Furry friends. PMID- 19189478 TI - Safety blankets. PMID- 19189479 TI - Pets win prizes. PMID- 19189480 TI - Individual budgets could at last lead to real changes in mental health services. PMID- 19189481 TI - Individual budget projects come under the microscope. PMID- 19189482 TI - It's time to talk. PMID- 19189483 TI - Taking it step by step. PMID- 19189484 TI - Changes to the Mental Health Act. AB - The revised Code of Practice reminds us: the patient's consent should be sought before treatment is given, wherever practicable. (Code, para 23.37) PMID- 19189485 TI - December is the festive season, but Rob Henderson has another take on the holiday period. PMID- 19189486 TI - Smoking has been banned in England's psychiatric hospitals--officially, at least. PMID- 19189487 TI - The SpmA/B and DacF proteins of Clostridium perfringens play important roles in spore heat resistance. AB - Strains of Clostridium perfringens that cause acute food poisoning have been shown to produce spores that are significantly more heat resistant than those of other strains. Previous studies demonstrated that the spore core density and the ratio of spore cortex peptidoglycan relative to the germ cell wall were factors that correlated with the heat resistance of a C. perfringens spore. To further evaluate these relationships, mutant strains of C. perfringens SM101 were constructed with null mutations in dacF, encoding a D,D-carboxypeptidase, and in the spmA-spmB operon, which is involved in spore core dehydration. The dacF mutant was shown to produce less spore cortex peptidoglycan and had a corresponding decrease in spore heat resistance. The spmA-spmB strain produced highly unstable spores with significantly lower core densities and increased heat sensitivity, which were easily destroyed during treatments affecting the spore coat layers. These results support the previous assertion that a threshold core density as well as a high ratio of cortex peptidoglycan relative to the germ cell wall contribute to the formation of a more heat-resistant spore in this species. PMID- 19189488 TI - Food allergies on the rise... PMID- 19189489 TI - Does head injury cause ADHD--or vice versa? PMID- 19189490 TI - Is giving acetaminophen rectally as good as giving it orally? PMID- 19189491 TI - New resources for clinical trials in children. PMID- 19189493 TI - Abstracts of the Second National Bioethics Conference. December 6-8, 2007. Bangalore, India. PMID- 19189492 TI - Unpasteurized milk--still a health threat. PMID- 19189494 TI - Special section: phylogenetics. PMID- 19189495 TI - Environmental analysis. PMID- 19189496 TI - [Snake bite in pediatrics: 6 years' experience of the University Hospital of Monterrey, NL]. PMID- 19189497 TI - Comment on "Serotypes and susceptibility of Streptococcus pneumoniae strains isolated from children in Mexico". PMID- 19189498 TI - Working with West African migrant communities on HIV prevention in Australia. PMID- 19189499 TI - Understanding oral sex. PMID- 19189500 TI - Introduction: drug trafficking and the state. PMID- 19189501 TI - Reflections on the changing status of academic science in India. PMID- 19189502 TI - Drugs in southern Africa: business as usual. PMID- 19189503 TI - Shift of preference of location for sexual partnership in men having sex with men in an Asian population. PMID- 19189504 TI - Unrecognised vitamin D deficiency: low concentrations in African migrants with HIV in Australia. PMID- 19189506 TI - [12th Congress on Atherosclerosis organized by the Czech Society for Atherosclerosis in cooperation with the Czech Diabetology Society CLS JEP. Spindleruv Mlyn, 11.-13. December 2008]. PMID- 19189505 TI - Money and the international drug trade in Sao Paulo. PMID- 19189507 TI - [Abstracts of the All-Russian Scientific Conference "Neurobiological Aspects of Morphogenesis and Regeneration" dedicated to the memory of the Corresponding Member of USSR Academy of Medical Sciences Professor F M Lazarenko. Orenburg, November 18-19, 2008]. PMID- 19189509 TI - Eternal dilemma: to take a stand or not. PMID- 19189508 TI - [In the memory of Natal'ia Petrovna Bekhtereva]. PMID- 19189510 TI - Lead your practice to success. PMID- 19189511 TI - Taking advantage of what is offered. PMID- 19189512 TI - Management of patients on bisphosphonates and prevention of bisphosphonate related osteonecrosis of the jaw. AB - Bisphosphonate related osteonecrosis of the jaw (BRONJ) is a recent discovery and has only been brought to the attention of health care professionals in since 2003. The purpose of this article is to describe this phenomenon and focus on preventive strategies to minimize the risk of patients developing this condition. Management and prevention will be broken down into patients taking IV bisphosphonates and patients taking oral bisphosphonates. PMID- 19189513 TI - Viewing movie smoking undermines antismoking parenting practices. AB - To test the hypothesis that viewing depictions of smoking in movies makes adolescents less responsive to parenting factors that prevent smoking. Cross sectional survey of 4807 students (grades 5-8) through which we ascertained exposure to smoking in movies, parent smoking, and adolescents' perception of parental responsiveness (support), and parental demandingness (behavioral control). Adolescents attending randomly selected middle schools in the Northeastern U.S. ever tried smoking a cigarette. Exposure to movie smoking was ascertained by counting occurrences of tobacco use in 601 recent popular motion pictures; surveying students to identify films they had seen from a random subset of 50 films; and summing tobacco use occurrences for the films each adolescent reported seeing. We also measured adolescents' perceptions of parent smoking, parental responsiveness and demandingness. The overall prevalence of adolescent smoking was 17.4 percent. The prevalence of smoking increased with exposure to movie smoking (low vs. high exposure 8.8 vs. 25.8%, p < 0.0001). Parenting factors associated with lower rates of adolescent smoking were parent non smoking status (11.0% vs. 27.7% for parents who smoke, p < 0.0001), higher levels of demandingness (13.7% vs. 20.7% for low demandingness, p < 0.0001) and higher levels of parental responsiveness (12.4% vs. 23.1% for low parental responsiveness, p < 0.0001). Parenting factors were not strongly associated with exposure to movie smoking. For adolescents with low exposure to movie smoking the adjusted odds (95% confidence interval) of smoking were 0.31 (0.23, 0.42) if parents did not smoke, 0.57 (0.42, 0.78) if parents exerted high demandingness, and 0.52 (0.38, 0.71) if parents were highly responsive. Parents had significantly less influence for adolescents with high exposure to movie smoking, for whom the adjusted odds of smoking were only 0.50 if parents did not smoke (p = 0.014 for the interaction effect), 0.97 if parents exerted high demandingness (p = 0.007 for the interaction effect) and 0.73 if parents were highly responsive (p = 0.045 for the interaction effect). Viewing smoking in movies not only has a direct association with adolescent smoking, it may also undermine parental attempts to prevent this behavior. PMID- 19189514 TI - [Smoking amongst women arriving at an outpatient clinic in hospital. Nicotine addiction versus disorders in the menstrual cycle]. AB - It is common knowledge that smoking badly influences women's health. Nicotine addiction does not only increase the risk of cancerous diseases, blood circulation and coronary system disorders as well as osteoporosis, but also it can cause disorders of the menstrual cycle, infertility and earlier menopause. One hundred and seven patients took part in the research and attended the women's clinic (Clinic "K"). On the basis of the questionnaire, some data was assessed: demographic data, frequency and type of menstrual cycle disorders, fertility and age of menopause occurrence. A group of smoking women (36.05%) was distinguished and non-smoking women (63.95%). A higher percentage of menstrual cycle disorders was noticed in a group of smoking women (p < 0.05). The most frequent symptoms were painful period, painful breasts and premenstrual tension. No significantly statistical difference was stated in frequency of menstrual cycle depending on the amount of smoked cigarettes and addiction duration. The main decisive factor of giving up smoking is the awareness of the harmfulness of nicotine addiction. PMID- 19189515 TI - [Influence of tobacco smoking on newborn's birth weight--analisys of dates concerning births from Maternity Hospital named. Dr S. Mossor's in Opole City]. AB - Despite wide education, tobacco smoking while being pregnant is very important issue in perinatology. It is important problem because of life style of polish society, including pregnant women. Clinical observation of this issue is pointing on risk of occurring pathology in pregnancy, unfavorable consequences for neonate also many distant pathological effects among children. Purpose of this was getting an answer for question: whether in current social and economic situation there is connection between low birth mass and smoking tobacco during pregnancy. Under analysis were found births between 38th and 40th one hundred successive births (according to book of birth-room from 2860 labors in hospital in Opol, 2007) of mothers are smoking up to 10 cigarettes a day (group I), mothers smoking 11-20 cigarettes a day (group II) and mothers that are not smoking. This works affirms that smoking has negative influence on child birth mass. It is also displayed that higher the number of smoked cigarettes the higher percent of newborns with low birth mass and higher number o fetus with intrauterine growth retardation. Among mothers that are smoking the biggest group were young women (mean. 24, years) and multipara female (58%). PMID- 19189516 TI - [Influence of smoking cigarettes on the development of the child and its health condition]. AB - 35.5% of Polish society is active smokers and as many as 20% of pregnant women. Influence of passive smoking on fetuses and children is bigger than active smoking on adults. The aim of this scrutiny was to develop relationship between preschool children passive smoking and their health condition. Research was made with the use of 100 anonymous questionnaires among parents of children from nursery schools in Opole. We received the following results: active smokers lived in almost half of examined families (44%), the most numerous group of smokers was parents (80.5%). The most often (43.3%) children were exposed on 20 cigarettes per day. 40.9% parents smoked outside, on the balcony, however the rest of them inside, in various rooms of the house. Most of parents thought that the health condition of their children was good, but more smokers than non-smokers estimated it as bad (11.4% vs. 1.8%). To the question how often are their children sick the most parents in both groups answered that 1-3 times per year. Most frequent illnesses concerned children of smokers and it was mainly respiratory tract diseases. Antibiotics intake was more often among children of smokers than non smokers. Relationship between their children health condition and cigarette smoking confirmed 90% of parents. Denied it 10% and all of them were smokers. It indicates the need of better education of both, parents and children, in the field of harmful effects of passive smoking on health. PMID- 19189517 TI - [Antismoking counseling]. AB - Smoking plays an important role in no communicable diseases including cardiovascular diseases etiology. In recent years more attention has been paid to the role of physicians in smoking prevention and treatment. The aim of this study was to analyze the presence of antismoking counselling given by physicians among the patients with the risk factors for cardiovascular diseases. The study was performed in the randomly selected group of 1267 men and women aged 18-65 years from Lodz and Lublin district. Each surveyed person had medical examination, blood pressure measured, anthropometry, biochemical tests included a full lipidogram and glucose concentration. Moreover detailed questionnaire surveys were conducted. Logistic regression models were applied to assess role of factors influencing undertaken of antismoking counseling among the patients with risk factors of cardiovascular diseases. The study revealed that smoking addiction affected 44% males and 23 % females. Abdominal obesity frequency in males was 24% and in females 35%. Hypertension was diagnosed in 30% of males and 21% of females, hyperlipidemia was diagnosed in 29% men and 19% women. High glucose concentration was identified in 7% of men and 5% of women. Only half of the smoking population received anti-smoking counselling given by physician. More frequently antismoking advice was given to the patients with abdominal obesity, (68%), and hypertension (67%). Only 36% of people with high glucose concentration received antismoking advice. This study results revealed, that anti smoking treatment among adults should be consider as an important part of preventing strategies. More effort should be given to increase awareness of physicians relating to the smoking habit of their patients. PMID- 19189518 TI - [Smoking and other risk factors of cardiovascular diseases, connected with arteriosclerosis among youth]. AB - The World Health Organization (WHO) conference on a "second wave" epidemic of cardiovascular diseases connected with arterial sclerosis (AS) foresee that in 2020 cardiovascular diseases will most likely be the leading cause of death in the world. The development of AS begins in youth and progresses with age. It's intensity depends on the risk factors involved, such as: smoking, hypertension, obesity and fat and sugar disorder in a body. Many of these risk factors, manifesting themselves as diseases in adults, can be found during adolescence. The aim of this study was to establish the spread of smoking and other risk factors of cardiovascular diseases, like: hereditary and increasing incidence hypertension and body mass index (BMI), among youth of upper gymnasium school in Podkarpacie. The research was conducted between November 2007 and March 2008, using 193 volunteer students from upper and lower gymnasium schools, aged between 16-20 years. Our research methods included: diagnostic questionnaire, measurement of blood pressure (BP) through the use of sphygmomanometer, as well as anthropometric measurements including high, weight and body mass estimation. BP was established by obtaining an average between two measurements taken under normal conditions. The results were statistically analyzed, in with the in dependent test chi-Parson square, the level of changes a = 0.05--was used. The research showed that 23.31% of respondents smoke, that's 64.44% girls, and 35.56% boys. 12.41% of the girls and 15.09% of boys smoke on regular basis. And 8.57% girls and 15.09% boys smoke from time to time. More than half of young smokers (51.10%) smoked for longer than 2 years, and the initiations of smoking starts at the age of 15 (26.67%) and the age of 16 (26.67%). 10 and more cigarettes a day smoke 26.67% of boys and 13.79% girls. 75.74% of respondents agree that they are victims of passive smoking. Through 17.61% of respondents (mostly boys 64.70%) we found increasing incidence of hypertension, and 82.35% were related to systolic BP. In the group of people with higher BP systolic hypertension demonstrated itself in 35.72% of positive cases, while diastolic hypertension related to 16.66% of the population and was present mainly among adolescence girls. Most of respondents with higher systolic (75.00%) and diastolic (83.00%) BP were found to be present in those with an obesity problem (50.00%, 10.00%) than in those with correct BMI (12.91%, 5.81%).Hereditary risk factors of AS, from father side, was found among 33.67% of respondents, and mother side, through 23.31% of respondents. Through respondents parents we found quite often: hypertension (fathers 18.65%, mothers 10.36%) and overweight and obesity (fathers 15.03%, mothers 13.99%) also through fathers we found hiperlipidemia (14.51%). Through mothers only, we found cases of diabetes (2.07%) but we didn't find heart stroke cases, which were found through fathers only (2.07%). Among respondents, 43.52% cases, we found one of risk factor of AS and more than half of respondents (56.48%) we found co -existence of 2 and more factors, including: 30.57%--2, 19.18%--3, 5.70%--4, 1.03%--5 risk factors. Through all respondents we found the existence of at least one of the risk factor of AS and through more than half of them, co-existence of two and more risk factors. Hereditary (33.67%--father side, 23.31%--mother side) and smoking (23.31%) were the common risk factors of AS in youth. An increase of hypertension and an increase incidence of BMI were present in (17.61%, 12.43%) respondents. PMID- 19189519 TI - [Tobacco smoking as a risk factor for oral cancer]. AB - The study involved 254 patients with primary oral cancer. The analysis of prevalence, duration and intensity of smoking habit was carried out both in cases and control group. The prevalence of smoking was significantly more frequent in cases than in controls. Any significant differences between cases and controls in duration of the habit and in number of cigarettes smoked daily were not noted. The findings indicated significant dependence between the smoking habit and the prevalence of oral squamous cell carcinoma. PMID- 19189520 TI - [Smoking and periodontal disease--results of an epidemiological survey]. AB - A questionnaire concerning oral health designed for young people aged between 18 and 30 and based on one produced by Tokyo University entitled Medical Outcome Study (MOS) and Oral Health Score) 8020, was submitted to a group of Poznan citizens. One of the 10 questions in the OHS part of the questionnaire concerned smoking habits. A condition for participation in the survey was an absence of professional or educational connections with medicine, dentistry or pharmacy. 506 persons (398 women and 198 men) living in Poznan, with an average of age 20.1 years took part. For a clearer evaluation of the health habits in this group, an additional comparison was made between the youngest (18 y old) and oldest (25-30 y old) groups. The Oral health Score (with a possible maximum 20 points) for the examined group was 14.4 which is one of the lowest among evaluated European countries. Smoking was most frequent among the 18 year olds (20.7%) whereas among the respondents in the older group it was approximately only half of this value (11.29%). Moreover in, both age groups, women were more often smokers than men; 22% of 18 year old women smoked and 7.5% in the older group whereas in younger men 7.5% smoked and 3.8% in the older group. Comparing the answers relating to oral health with the frequency of smoking revealed a connection between this habit and the presence of periodontal disease. PMID- 19189521 TI - [The tobacco smoking and basic coefficients of nutritional status of youth in secondary schools]. AB - The aim of the study was the assessment of cigarette smoking by youth from secondary schools. The study was conducted among 700 pupils of secondary schools from Poznan in the year 2004 and it was based on questionaire survey regarding frequency and the period of smoking, as well as the basic anthropometric measurements including body mass and height. The study revealed notable proportion of girls with underweight, however the boys were mostly overweight. Smoking habits concerned often girls than boys. However higher proportion of boys started earlier smoking comparing to girls. Both girls and boys have smoked cigarettes not earlier than during last five years. It was proved that most pupils were not pharmacologically addicted to nicotine. Notable number of smokers tended to quit smoking. PMID- 19189522 TI - [Tobacco smoking and frequency of arterial hypertension in students of Poznan universities]. AB - The aim of the study was to assess the tobacco smoking among students of three Poznan universities and its effect on the students' blood pressure. Into the consideration were taken the frequency and duration of tobacco smoking, and the number of cigarettes smoked daily. The study involved 700 students. An appropriate questionnaire was used to collect the data, and blood pressure measurements were taken. The results were statistically analyzed using t-Student and chi2 tests. The results show that the gender and type of the university attended influence the prevalence of tobacco smoking. The percentage of smoking men was significantly higher than the percentage of smoking women. The lowest percentage of smokers was found among the students of the Medical Sciences University as compared with other universities. In addition, some positive correlation between tobacco smoking and the occurrence of arterial hypertension was observed. PMID- 19189523 TI - [Structural changes in tracheal epithelium in environmental smoke exposed rats- experimental studies]. AB - It has been already proved in many experimental studies that tobacco smoke has multiple toxic effects on respiratory tract cells. Alterations in cilliary epithelium of rats trachea after short exposition to high tobacco smoke concentrations in inhaled air were been determined in current study. Morphological evaluation revealed in lining epithelium decrease in number of cilliary cells and increase in number of goblet cells. The mucous membrane was thickened and infiltrated with inflammatory cells. PMID- 19189524 TI - [Distribution of alcohol dehydrogenase (ADH1C) genotypes in subjects with tobacco smoke-associated laryngeal cancer]. AB - Laryngeal cancer in Poland is characterized by high levels of morbidity and mortality. The main risk factors for the larynx cancer are alcohol drinking and tobacco smoking. In contrary to well established tobacco-related evidence for an increased risk of larynx cancer, alcohol-related mechanisms of carcinogenesis remain unknown. Nevertheless the effect of alcohol is modulated by polymorphisms in genes encoding enzymes for ethanol metabolism. Hence we investigated the ADH1C *1 genotype and allele frequency in a group of 102 larynx cancer patients with heavy alcohol consumption recruited from the Department of Otolaryngology and Laryngological Oncology of the University of Medical Sciences in Poznan. The data were compared with 112 non-cancer age-matched individuals consuming similar amounts of ethanol. Blood samples were used for analysis of restriction fragment length polymorphism. DNA was isolated from the whole blood leucocytes and PCR with specific primers was used to amplify polymorphic region of rs698 in the ADH1C gene. The method was based on allele detection by Sspl restriction enzyme digestion and after the incubation with enzyme, samples run on an electrophoresis. The statistic analysis was performed to calculate Odds Ratio (ORs), 95% confidence intervals (CIs) and significance. Results suggest a slightly increased risk of larynx cancer for individuals who have inherited the ADH1C *1 allele (rs 698), however they did not reach the level of statistic significance. PMID- 19189525 TI - [The influence of lead on concentration of the pregnancy-associated plasma protein A (PAPP-A) in pregnant women smoking tobacco--preliminary study]. AB - The epidemiology studies conduct in Institute of Mother and Child indicated that in Poland 25-30% women smoke during pregnancy. Lead exposure from cigarette smoke may have a negative effect on transplacental transpire of micronutrients and if coexists with low concentration of pregnancy. associated plasma protein A (PAPP A) resulting in adverse influence on growth and fetal development. Literature date points, those clinical symptoms of high blood level of lead and insufficiency of PAPP-A are similar. Therefore the aim of the study was to estimate the effect of lead from cigarettes on plasma PAPP-A levels in smoking pregnant women. Healthy, pregnant women (n = 30), patients of Clinical Department of Obstetrics and Gynaecology, Institute of Mother and Child were divided into groups nonsmoking and smoking according to questionnaire declaration. Plasma concentrations of lead were analysed using inductively coupled plasma mass spectrometry (ICP-MS) on spectrometer analyser ICP MS Elan 6100 (Perkin Elmer, Germany). Plasma level of PAPP-A was measured by immunofluorescence method using BRAHMS (Germany) kits on KRYPTOR (BRAHMS, Germany) analyser. We observed that in plasma of smoking pregnant women the concentration of lead was 2.55 microg/l (range: 0.78-14.6 mg/l) and was higher by 30% than in tobacco abstinent (1.78 microg/l; range: 0.0-4.69 microg/l). In group of smoking mothers level of PAPP-A was lower by 20% than in nonsmoking ones (2.80 IU/L; range: 1.3-9.3 IU/L vs 3.2 IU/L. range: 1.7 - 6.8 IU/L). Our results indicate that tobacco smoking during pregnancy affect in maternal blood lead and plasma protein A values. In smoking women lower concentrations of PAPP-A co-exist with higher than in tobacco abstinent level of lead which may suggest correlation between these parameters. Further studies will be continued in larger group of women in all trimesters of pregnancy to confirm of this association. PMID- 19189526 TI - [The effect of tobacco smoking during pregnancy on concentration of pro-hepcidin and some parameters of iron metabolism in matched-maternal cord pairs]. AB - Iron deficiency relatively observed in pregnant women is assumed to be enhanced by cigarette smoking. Hepcidin, a peptide hormone produced by the liver as pro hepcidin, has recently emerged as a central mediator of iron metabolism. Hepcidin regulates intestinal iron absorption, macrophage iron release, and the placental passage of iron. Maternal smoking is associated with increased fetal iron requirements and stimulates fetal erythropoiesis. This is probably through a hypoxic effect on the fetus, and is dose related to the maternal smoking level. It is known that anemia and hypoxia suppress hepcidine mRNA expression. Therefore the aim of the study was to estimate the effect of tobacco smoking on serum pro hepcidin levels and some iron parameters in pregnant women and umbilical cord blood. We also studied correlation between pro-hepcidin and others iron markers in mothers and their newborns. Healthy, pregnant women (n = 50), patients of Clinical Department of Obstetrics and Gynecology, Institute of Mother and Child were divided into groups nonsmoking and smoking according to questionnaire declaration. Serum concentrations of pro-hepcidin were determined by immunoenzymathic method using a commercial pro-hepcidin assay (DRG, Germany). Levels of ferritin and transferrin were measured by immunoturbidimetric method and iron by photometric test with ferrozine using HORIBA ABX kits (France) and Cobas Mira analyser (Roche, Switzerland). Levels of hemoglobin and hematocrite were determined using commercially available kits on Pentra 60 analyser (ABX, France). We observed that the mean concentration of pro-hepcidin in serum of smoking pregnant women was statistically lower than in tobacco abstinent (101.9 +/- 28.6 ng/ml vs 88.3 +/- 18.2 ng/ml; p < 0.01). Levels of others studied iron markers were similar in both group except total iron concentration, which was 20% lower in smoking mothers than in nonsmoking ones. In umbilical cord blood of infants born to smoking women level of pro-hepcidin was significantly lower than in tobacco abstinent (54.2 +/- 14.0 ng/ml vs 76.8 +/- 21.4 ng/ml, p < 0.0001). We observed positive correlation between concentrations of that prohormone in serum of mothers and cord blood of their newborns in nonsmoking group (r = 0.54; p < 0.02) as well as in smoking ones (r = 0.68; p < 0.05). In addition, concentrations of ferritin, transferin and total iron were lower by 30%, 13% and 20% respectively in cord blood of smoking than nonsmoking group. The differences were statistically significant (p < 0.05). Our analysis revealed no correlation between serum pro-hepcidin levels and other studied parameters of iron status both in the mothers and children groups. Our results indicate that tobacco smoking during pregnancy affected pro-hepcidine levels in serum of mothers and their newborns. Low concentrations of some iron markers in umbilical cord blood suggest that mother's smoking could lead to subclinical iron deficiency in fetus. No anemia were observed in both studied groups of mothers that could explain no relationships between pro-hepcidin and others parameters of iron status. PMID- 19189528 TI - [Tobacco smoking and biochemical acute phase markers among the patients with chronical periodontal disease]. AB - The periodontal status of 34 patients age 32 to 60 years, including 8 smokers, was evaluated. It was based on the medical history, progress of periodontal disease measured by probing pocket depths and attachment loss, oral hygiene status and presence of gingivitis. Analysis of acute phase proteins and total antioxidant status as a response to chronic inflammation in the oral cavity were performed by means of immunelectrophoresis according to the Laurell and Randox tests. The result suggest that the effect of a bacterial biofilm, deepened pockets and smoking is more important for development on the pathological process than genetic factors and gender. PMID- 19189527 TI - [Effect of maternal smoking on concentrations of the pregnancy-associated plasma protein A (PAPP-A) and free beta subunit of chorionic gonadotropin (beta hCG) in the first trimester of pregnancy]. AB - A combination of maternal serum levels of free beta human chorionic gonadotropin (free beta hCG) and pregnancy-associated plasma protein-A (PAPP-A) and fetal nuchal transluency thickness (fetal NT) has been shown to be an effective approach to screening for fetal trisomies in the first trimestr of pregnancy. A maternal smoking habit is known to influence these serum markers in singleton pregnancies however it has not been yet completely explained. The aim of this study was the investigation of PAPP-A and beta hCG concentrations in smoking women in the first trimester of pregnancy. The study comprised 215 women between 11th and 14th weeks of gestation: 135 cigarette smokers (age range 19-43 years) and 80 nonsmokers (age range 21-42 years). Only normal singleton pregnancies were included. Smoking women were divided into two groups: 75 women smoking during all along the first trimester and 60 women, who stopped smoking before 6 week of gestation. Maternal serum levels of PAPP-A and beta hCG were measured with the KRYPTOR rapid random-access immunoassay analyser (Brahms GmbH, Germany) using time-resolved amplified cryptate emission technology (TRACE). Combinated fetal nuchal translucency (NT), maternal age and biochemical risk estimates were calculated using the Fetal Medicine Foundation risk algorithm. We observed the lower of PAPP-A median values in both smoking groups by about 20% than in nonsmokers (0.86 MoM; 0.84 MoM versus 1.04 MoM; p < 0.01). We also indicated the lower values of beta hCG in smoking women in comparison to the nonsmoking controls, but this difference was not statistically significant. The concentrations of PAPP-A as well as beta hCG were similar in both groups of smoking women. Further studies will be continued to assessment of smoking influence before conception on maternal serum PAPP-A and beta hCG in the first trimester of pregnancy. PMID- 19189529 TI - [Hiperhomocysteinemia in active and passive smokers and the levels of folate and vitamin B6 in plasma]. AB - The increased plasma level of homocysteine have been shown to be the sensitive marker for the folate, vitamin B6 and cobalamins deficiency and an independent risk factor for the cardiovascular disease, neutral tube defects and a potential causal risk factor for neuropsychiatric disorders. The blood and plasma homocysteine levels except for genetic defects are influenced by age, gender, efficiency of detoxication systems, one or more unhealthy lifestyle factors, such as high alcohol consumption, low nutritional intake of vitamins, high coffee consumption, acquired disorders and lack of physical exercise. Many studies confirm that active tobacco smoking and environmental tobacco smoke (ETS) have been significantly associated with hiperhomocysteinemia. In metabolic pathway of homocysteine the important role played folic acid, as a donor of methyl group in re-methylated reaction to methionine and vitamin B6. It acts as the cofactor in transsuphuration reactions of homocysteine to cystathionine and cysteine. Hence, the aim of this work was to compare the plasma folate and vitamin B6 concentrations in smokers and passive smokers with a hiperhomocysteinemia (> 15 micromol/L). It was observed that the plasma folate levels in active (n = 30) and passive smokers (n = 29) groups decrease statistically significant (P < 0.001) in comparison to non-smokers (n = 37). The calculated Spaermann's correlations coefficient of total plasma homocysteine level and plasma folate concentrations in the non-smoker group indicated a weak, statistically insignificant correlation (r = -0.103, P = 0.542). However, the above relationship in passive and active smokers were statistically significant (r = -0.495, P 0 0.01; r = -0.672, P < 0.001, respectively). The decrease of vitamin in B6 plasma was observed in all active smokers group (P < 0.01) and men smokers comparing to non-smokers (P < 0.001). There was no observed significant correlations between hiperhomocysteinemia and vitamin B6 in all studied groups. The results indicated that hiperhomocysteinemia have strong negative impact on folate levels in active and passive smokers. The tobacco smoke exposure have negative influence on the status of vitamin B6. The lack of significant correlation between increased homocysteine levels and vitamin B6 status confirmed hypothesis that hiperhomocysteinemia is not depended on vitamin B6 concentrations in plasma. PMID- 19189530 TI - [The activity of thermolability amylase in serum nonsmoking and smoking healthy persons and patients with pancreatitis]. AB - The aim of this study is to prove the influence of tobacco smoking on total and thermolability amylase activity in the serum of non-smoking and smoking health persons and patients with diagnosed acute (AP), chronic exaggerated (CEP) and chronic pancreatitis (CP) and patients with diabetes. The blood has been collected from 28 healthy persons and 52 patients. The enzyme total activity has been determined using the colorimetric method with substrate 1,2-odilauryl-rac glycero-3-glutaric acid -(6-methylresorufin) ester. The thermolability activity has been determined using the thermolability test. The tobacco smoke has been examined on the basic of concentration of cotinine in the serum of health persons and patients. The highest amylase total activity and her thermolability form have been found in smoking patients with diabetes. It has been noted that the serum amylase activity is significantly higher in smoking and healthy persons (p < 0.0002; p < 0.002) then in non-smoking and healthy patients. However no significant differences have been found between the thermolability total activity, however it has been noted higher thermolability thermolability activity in smoking patients with CP and nonsmoking patients with CP. Smoking patients with AP and CEP have been found to have a significantly increased enzyme and her form thermolability activity (p > 0.001; p > 0.005 respectively) when compared to non-smoking patients. Results of examination indicate that tobacco smoking has a significant influence on pancreatic amylase activity. PMID- 19189531 TI - [Influence of tobacco smoking on amylase activity in serum persons occupational exposed to heavy metals]. AB - The newest conducted investigations showed the significant role of tobacco smoking in inducing pathological changes in pancreas. Additionally exposure to heavy metals presents on polluted environment influences on function this organ. However, the mechanism of development of these changes has not been fully recognised. The aim of this study is to prove the influence of tobacco smoking on total amylase and termolabile amylase activity in serum of smoking and nonsmoking healthy persons and workers at cooper foundry in Legnica occupationally exposed to heavy metals: cadmium, arsenic, lead. Blood has been collected from 28 healthy persons and 60 founders. The enzyme total activity has been determined using the colorimetric method with substrate 1,2-odilauryl-rac-glycero-3-glutaric acid-(6 methylresorufin) ester. The thermolability activity has been determined using the thermolability test. It has been noted significant higher total amylase and thermolabile amylase activity in serum of smoking healthy persons (p < 0.0002; p < 0.002) and of non-smoking (p < 0.001; p < 0.01) and smoking founders (p < 0.0004; p < 0.001) comparison with non-smoking healthy persons. It hasn't been found significant differences in total and thermolabile amylase activity in smoking founders and non-smoking founders. The fact that there are significant differences in serum amylase activity in serum of smoking and nonsmoking founders in comparison with nonsmoking healthy persons prove a significant influence of exposure to heavy metals on exocrine function of pancreas. PMID- 19189532 TI - [The influence of smoking of the cigarettes on the level of circulating immunological complexes in the serum of patients with the hypersensitivity of the type I]. AB - Circulating immunological complexes are the answer of the immunological system on appearing in the organism of the substance about the character of antigenic. The growth of the concentration of immunological complexes is described in many diseases among others in allergies. The aim of the work was the opinion of levels of circulating immunological complexes serums smoke patients and no smokes with the hypersensitivity of I type. Patients serums given an examination with the hypersensitivity of the type I (n = 22) smoking (n = 11) and nonsmoking (n = 11). 45 persons been careful for healthy made up the supervisory group. The level of circulating immunological complexes was studied the simple method of PEG precipitation according to the Haskov method. Smoking patients step out higher levels of CIC in the hypersensitivity of the type I than at patients who don't smoke. PMID- 19189533 TI - [Tobacco smoke exposure levels and lipide profile]. AB - Toxic tobacco smoke exposure to human organism is strictly related to progress of atherosclerosis changes. One of the mechanisms of these effects is a change of blood lipoprotein fraction concentrations. The concentrations of the lipid profile parameters (TCL, HDL, LDL, TG) and the chosen biomarkers (urine cotinine and 1-hydroxypyrene and blood carboxyhaemoglobine) were determined. It was studied whether the procedure of the groups determination (nonsmokers, passive and active smokers) affect the calculated average values of lipid profile parameters. The role of the applied biomarkers for the detection of the effects related to the tobacco smoke exposure is also discussed. It is concluded that there is no difference among the lipid profiles of passive smokers and nonsmokers. Cigarette smoking increases and lowers the TG and HDL concentrations, respectively. Urine cotinine seems to be the best indicator of tobacco smoke exposure among three chosen biomarkers. Study subjects were 300 male and female volunteers. PMID- 19189534 TI - [The influence of smoking during pregnancy on decresed vitamin C concentration in blood serum]. AB - There are reports about decreased vitamin C concentration in blood serum of smokers. In pregnancy decreased vitamin C concentration in blood serum is observed as well. The aim of this study was to analise vitamin C concentration in blood serum of pregnant women in two periods of pregnancy (20-22 and 38-41 week), in smokers and non-smokers and its comparision to our own results from researches made 25 years ago. Vitamin C concentration was determined by using Roe and Kuether method, in groups of 20 prgnant women being in 20-22 and 38-41 pregnancy weeks, smoking over 10 cigarettes per day and non-smoking. Because most of pregnant were using recommended vitamins, only these were qualified for research, who were taking multivitamin supplements including vitamin C in daily dose from 100 to 180 mg. Statistical analysis of the results was made by using t-Student test with changeability for p < 0.05. RESULTS: average vitamin C concentration in pregnant woman blood serum smoking over 10 cigarettes per day in 20-22 week of pregnancy was 1.43 mg% and in 38-41 week was 1.35 mg%. This concentration is much higher than that compatible from 25 years (0.51 mg% in both 20-22 and 38-41 week of pregnancy). Vitamin C concentration in non-smoking pregnant woman blood serum are higher now as well. These diffrences should be explained by taking multivitamin supplements widely recommended during pregnancy, by education and promotion of healthy living, by improvement of socioeconomic and environmental conditions. However the fact of harmful influence of cigarette smoking on vitamin C concentration in pregnant woman blood serum is still clear. PMID- 19189535 TI - [Tobacco smoking influence on the level of sex hormones--animal model]. AB - Numerous studies warn that women who smoke can suffer from weakened functioning of their ovaries and disturbed synthesis and metabolism of hormones. This may cause many pregnancy complications or premature menopause and osteoporosis. Moreover, smoking disturbs the menstrual cycle, decreases the effectiveness and increases the undesirable effects of the hormone replacement therapy. Tobacco smoke disturbs gametogenesis, ovulation, Fallopian tube transport, fertilization and the implantation of a fertilized cell, which results in the reduction of fertility. The goal of the present thesis was a toxicological assessment of the influence of tobacco smoke on the level of luteinizing hormone, follicle stimulating hormone, progesterone, and estradiol in blood serum of female rats, taking into consideration the phases of their sexual cycle. The experiment utilized a cytological method of determining the phases of the sexual cycle, which enabled the researcher to divide the rats into two groups (exposed to tobacco smoke and unexposed). Each of the groups was further divided into four subgroups with six animals for each phase of the sexual cycle (Proestrus, Estrus, Metestrus, Diestrus). The rats from the first group were exposed to tobacco smoke with the concentration of 1500 mg of Carbon Monoxide (CO) per cubic meter of ambient air (per content of Carbon Monoxide) for 5 days for 6 hours a day. Every day before the exposition at the same time the phases of the sexual cycle of female rats were checked in all of the subgroups. On the last 5th day of the experiment, after the exposition to the smoke, smear tests were conducted. The unexposed group was the control group. The evaluation of the exposition of the animals to tobacco smoke was based on the determination of the level of cotinine in blood serum by ELISA method. The measurement of the concentration of hormones was conducted by means of a jurisdiction method of electrochemiluminescence. In the experiment, a statistical difference was observed between the concentration of cotinine in the blood serum of the animals that were in the Proesrtus phase (86.8 ng/ ml), compared to the group in the Metestrus phase (351.0 ng/ml) and Diestrus phase (304.6 ng/ml). In the experiment a very high level of progesterone concentration was marked in the blood serum of the female rats that were exposed to tobacco smoke and that were in the Proestrus phase (195.1 ng/ml). The level of progesterone among the animals exposed to cigarette smoke was statistically higher compared to the control group (99.1 ng/ml). In all probability, this was caused by the slower metabolism of nicotine or faster biotransformation of cotinine. The experiment confirmed the negative influence of cigarette smoke on the course of the sexual cycle. The sexual cycle of the female rats exposed to tobacco smoke changed its rhythm and kept its proper course only in 29% of the examined animals. PMID- 19189536 TI - [Evaluation of selected endocrine parameters in smoking patients with arterial hypertension]. AB - Among many actions of tobacco smoke very important is: metabolic influence and other biological effects e.g. hormones secretion. Results of research prove the influence of smoking on pancreatic and adrenal secretion and on the largest endocrine organ of human body - endothelium. The aim of the study was evaluation of selected endocrine parameters: insulin and aldosterone (Ald) concentration, plasma renin activity (ARO) and endothelin-1 concentration (ET-1) in smoking patients with arterial hypertension in comparison with hypertensive non-smokers. The examined group comprised 27 patients with essential hypertension (grade I and II). The group was divided into 2 subgroups: Subgroup A--smoking hypertensive patients (10 persons). Subgroup B--non-smokers with hypertension (17 persons). The significant differences between 2 subgroups were noticed as for the following parameters: a) insulin concentration - Me +/- P: in subgroup A: 20.75 +/- 13.4 v. in subgroup B: 10.5 +/- 9.67 microIU/ml, p = 0.023; b) Ald - Me +/- P: in subgroup A: 280 +/- 155.3 v. in subgroup B: 54.95 +/- 124.33 pg/ml, p = 0.045. The differences between 2 subgroups were non-significant as for the following parameters: c) ARO - Me +/- P: in subgroup A: 3.85 +/- 1.22 v. in subgroup B: 1.8 +/- 1.61 ng/ml/h, p = 0.127; d) ET-1- Me +/- P: in subgroup A: 58.58 +/- 23.14 v. in subgroup B: 59.93 +/- 12.55 pg/ml, p = 0.901. Tobacco smoking affects many hormonal parameters. In patients with essential hypertension we observed significantly higher insulin and aldosterone concentration in the subgroup of smokers in comparison with nonsmokers. PMID- 19189537 TI - [Urinary N-acetyl-beta-D-glucosaminidase and its isoenzymes in smoking and non smoking workers at copper foundry occupational co-exposed to arsenic cadmium and lead]. AB - The kidneys are the critical organs in the case of a long-term occupational or environmental exposure to heavy metals and tobacco smoke. In diagnostics of renal damage useful are the methods which determine the activity of renal enzymes, quantify in urine (e.g. beta-glucuronidase, N-acetyl-beta-D-glucosaminidase). N acetyl-beta-D-glucosaminidase (NAG) is one of the most often determined factors of tubular damage, since its activity increases in early stages of renal injury, ahead of appearance of excretory dysfunction. The aim of this research was to assess the influence of occupational exposure of copper-foundry workers to heavy metals (arsenic, cadmium, lead) on total activity of N-acetyl-beta-D glucosaminidase and its molecular forms in urine. The investigated group was made up of 95 founders (smokers n = 51, non-smokers n = 44) and 43 people in control group (smokers n = 16, non-smokers n = 27). The concentrations of arsenic (As) and cadmium (Cd) were determined in urine, whilst the level of lead (Pb) was determined in whole blood. The activities of NAG and its isoforms were determined in urine. Smoking and non-smoking founders' urine demonstrated 14 times higher concentrations of arsenic levels in comparison with smoking and nonsmoking control group. Cadmium concentrations were 3.5 times higher in urine of smoking founders in comparison with smoking control group and about 3 times higher in case of nonsmoking founders in comparison with non-smoking control group. 7 times increase of lead concentration was observed in the whole blood within the smoking founders group in comparison with the smoking control group. In the blood of non smoking founders was demonstrated about 10 times increase of lead concentration in comparison with the non-smoking control group. About 3-times increase of total NAG's activity was observed in urine of smoking founders and 4-times increase of non-smoking founders in comparison with smoking and non-smoking control group. The highest activity of NAG-B was observed in urine of smoking founders (11.35 +/ 7.85 U/g creatinine), then non-smoking founders (9.7 +/- 8.75 U/ g creatinine). It was confirmed, that the activity of N-acetyl-beta-D-glucosaminidase is a good factor in the assessment of occupational exposure to heavy metals like arsenic, cadmium and lead. PMID- 19189538 TI - [Tobacco smoking by alcohol addicted patients--preliminary report]. AB - The problem of alcohol and nicotine addiction is both social and medical. The substances are often used together and give complex addiction. The knowledge about mechanisms of complex addiction would be useful for creating effective ways of treatment the abused patients. The aim of the paper was to describe the complex ethanol and nicotine addiction phenomenon. The research was made using data from 30 ethanol and nicotine addicted patients treated in Clinic of Toxicology Jagiellonian University Collegium Medicum in Krakow. 90% of them were nicotine addicted. Tobacco was the first abused substance in the group of patients. The mean period of addiction was 29.0 years for tobacco and 22.2 years for alcohol. The majority of the patients (51.9%) smoked more than 20 cigarettes per day; 29.6% smoked 11-20 cigarettes per day. Single patients were narcotics and drugs addicted. Continuation of the research and toxicological determination of the substances and their metabolites concentrations would be useful for the complex addiction problem explanation and description. PMID- 19189539 TI - [Chromium in hydroxiapatites of gallstones from active and passive smoking and non smoking women]. AB - The studies were carried out on the changes in chromium content in concretions of gallstones taken from 93 women (24 active smoking, 18 passive smoking, 51 non smoking women), respectively. Chromium content was determined with ICP-AES method (Inductively Coupled Plasma - Atomic Emission Spectroscopy). Studies on chromium occurrence in concretions of gallstones showed the presence of its higher concentrations in passive smoking women (3.15 microgCr/g) in comparison with active smoking (1.20 microgCr/g) and non smoking women (0.89 microgCr/g). Separate patterns of changes in chromium in active, passive and no smoking women gallstones were demonstrated. PMID- 19189540 TI - [The occurance lead and cadmium in hip joint in aspect of exposure on tobacco smoke]. AB - The objective of this study was qualification of content cadmium and lead in selected elements of the hip joint in aspect of tobacco smoking. The material for the research were 5 elements of hip joint (articular cartilage, trabecular bone and cortical bone femur head, fragment articular capsule and fragment trabecular bone from region intertrochanteric femoral bone), obtained intraoperatively during endoprothesoplastic surgeries. The samples come from habitants of Upper Silesian Region. Determination of trace elements contents were performed by ASA method (Pye Unicam SP-9) in acetylene-oxygen flame. Higher contents of lead were observed for smoking people, however in case of cadmium the differences of this element were not statistical essential between smokers and non-smokers. PMID- 19189541 TI - [Accumulation of Cu in pharyngeal tonsils from children exposed and unexposed to smoke of tobacco]. AB - The characteristic of occurrence Cu content in pharyngeal tonsils from girls and boys exposed to tobacco smoke and unexposed to tobacco smoke living in Southern Poland is presented in this article. Analysis of content of Cu in pharyngeal tonsils was observed in eight groups of children: girls exposed to tobacco smoke and unexposed to tobacco smoke living in industrial region and village, boys exposed to tobacco smoke and unexposed to tobacco smoke living in industrial region and village. Geometric mean of contents of copper in pharyngeal tonsils from girls exposed to ETS living in industrial region was 1.35 microg/g and no exposed girls was 1.31 microg/g, in girls exposed to ETS living in village region was 0.72 microg/g and no exposed to ETS girls was 0.66 microg/g. Geometric mean of contents of copper in pharyngeal tonsils from boys exposed to ETS living in industrial region was 1.76 microg/g and no exposed to ETS boys was 1.10 microg/g, in boys exposed to ETS living in village region was 0.71 microg/g and no exposed girls was 0.30 microg/g and this differences was statistically important. The cross-correlation analysis between content of Cu and other metals Ba, Ca, Cr, Al, Fe, Mg, Mn, Zn, K, Na in eight groups of children was observed. PMID- 19189542 TI - [Co-occurrence Fe in relationships to other metals in pharyngeal tonsil of children in view of sex, age and expose to passive smoking]. AB - The aim of this study was analysis of kind of interactions between Fe and the other metals: Al, Ba, Ca, Cr, Cu, Mg, Mn, Zn, K i Na in pharyngeal tonsils from girls and boys in several group of age (2-5 years, 6-9 years and 10-15 years) exposed and no exposed to tobacco smoking. This study showed that the kind of interaction dependent on sex, age and exposed to ETS. PMID- 19189543 TI - [Selected indicators of tobacco smoking in 15-year-old students in Poland in relation to international statistics]. AB - HBSC (Health Behaviour in School-aged Children) study can serve as a basis for monitoring smoking prevalence in school-aged children. Previous studies 1990-1998 have shown an increase in tobacco smoking among 15-year-old students, followed by a decrease in 1998-2002. The aim of this study was to present current international comparisons of selected indicators and changes in 2002-2006. An international standard questionnaire prepared in the frame of HBSC study was used and school-based cross-sectional survey was conducted. In Poland, the sample was selected as representative for pupils from III grade of lower secondary school with mean age of 15.7 yrs (2002 N = 2152; 2006 N = 2287). Polish data was compared with combined international samples from 37 countries. In 2006, 58.9% of Polish 15-year-old students reported that ever tried to smoke, 24.6% smoked at the time of survey, while 12.4% were regular smokers. The proportion of children who reported every day smoking has decreased substantially during 2002-2006. The rate of change was lower in girls than in boys. Decreasing trend was observed in most HBSC network countries. However, in most countries the improvement was higher than in Poland, especially in girls. More attention should be paid to divergent trends in smoking among adolescent boys and girls. Despite favorable trends in Polish boys, smoking prevalence is still too high by comparison to other countries and age of smoking initiation is very low. PMID- 19189544 TI - [The age of tobacco initiation and tobacco smoking frequency among 15 year-old adolescents in Poland]. AB - The aim of the study was to present the relationship between the age of tobacco initiation and current number of smoked cigarettes and frequency of smoking among 15 years old adolescents in Poland. The research was conducted in 2006 as a part of Health Behaviour in School-aged Children: A WHO Collaborative Cross-national Study (HBSC). The representative sample consists of 2152 pupils (1037 boys and 1115 girls) at the average age of 15.7. In analysis only the adolescents who have ever smoked tobacco were taken into account. For the research purpose the standard international HBSC questionnaire was used. The presented results show that adolescents who began smoking at young age smoke more cigarettes than equals, who started smoking later. These differences are especially pronounced in the group of girls. Conducted research indicates, that smoking tobacco 10 and more times in 30 days and 12 months before survey as well, occur statistically more frequently in a group of girls, who belong to early initiators. In the group of boys, smoking more cigarettes was related to late tobacco initiation. In smoking prevention actions conducted among the adolescents it is recommended to lead campaigns pro delaying the tobacco initiation age. The current smoking stage, the factors influencing tobacco consumption and gender of programs recipients should be taken under consideration as well. PMID- 19189545 TI - [Tobacco smoking by family members at home and adolescents' subjective health]. AB - The aim of the study was: (1) to examine frequency of tobacco smoking by family members in the opinion of 12-16-year-old adolescents (2) to investigate the relation between multiple subjective complaints and exposure to passive smoking at home. The results of surveys conducted among 778 pupils at the age of 12-16 in 2003 were used. The research took place in Poland in the year 2003. The article presents the estimation of the risk of frequent (most days or more than once a week) multiple subjective complaints in relation to the exposure to passive smoking at home, gender, age and family's financial situation. Among questioned adolescents 39% stated that the members of their families smoke in their presence, in 12% of the cases this situation happens very often or always. In the group of the young people exposed to passive smoking at home statistically more frequently occur single complaints like dispiritedness, exasperation or nervousness and as well multiple subjective complaints. Exposing adolescents to passive smoking at home is related to frequent occurrence of negative emotional states and multiple subjective complaints. Leading further campaigns against passive smoking is a necessity. PMID- 19189546 TI - [Smoking as an aspect of lifestyle among 18-year-old secondary school students in Poznan]. AB - Smoking remains a serious health threat in Poland. The age of 18 years is set as a conventional and symbolic boundary when young people enter adulthood and, at the same time, gain legal access to cigarettes and alcohol. At this age, however, most smokers have already developed smoking habits. This study seeks to identify how prevalent is smoking among 18-year old learning adolescents, and what is its relation to certain social factors and students' lifestyle. In our research, we used statistical methods and a questionnaire-based survey. The research was conducted in 2007 and it covered 400 third-year secondary school students in Poznan. Smoking among adolescents remains at a relatively high level and is related to the type of school, fulfilment of material needs, subjective perception of a state of health, use of other drugs, and a chosen lifestyle. PMID- 19189547 TI - [The reasons of smoking cigarettes among students in Poznan universities--a comparative study]. AB - Smoking cigarettes is one of the most common factor disturbing health and a part of negative life style. The aim of the study was to estimate the frequencies of smoking cigarettes and their conditions among students of two Poznan universities: Poznan University of Medical Sciences and Poznan University and three lines: physiotherapy, midwifery, psychology. 84 persons participated in the study. 22.6% of them were smoking and almost 80% were not smoking. 51% persons smoked in the past. Author's questionnaire of own concept used in the study. The research was conducted according to law of care about personal data. RESULTS: 9.5% smoking responders did it after breakfast, 16.6 % at working place during breaks. The data show that stress is a main reason of smoking. Almost 9.5% smoke during brakes at work and in social circumstances. According to opinion of examined group, the joy, relax are the effects of smoking. There were no significant differences and correlations between fear as a state and character and smoking. However the frequency of smoking in the group was lower in comparison to medium values in population of central part of Europe in a group of women. There was no correlation between sex and the frequency of smoking. The frequency of smoking in a group was lower in comparison to data from Central Europe region. PMID- 19189549 TI - [Tobacco smoking and self-assessment of health status among students from High School of Country Economy in Kutno--preliminary study]. AB - Tobacco smoking is still actual and common problem, which affects both students' high schools and their professors. In this study results are presented among students from one private schools in Poland, when students are educated in the following directions: geodesy, Europe science, pedagogy, computer science and nursing. The aim of the study was to assess prevalence of tobacco smoking among students and the awareness of health consequences. It is also decided to check which variables determinate self-assessment of health status of students and what motives of tobacco smoking are. Tobacco smoking was declared by 39% of students, 81.9% of them smoked regular and 18.1% - occasional. The biggest group of tobacco smoking students was noticed in geodesy - 35.4% students and nursing - 29%. Nearly 44% had opinion that tobacco smoking become addicted (22.9% students from nursing, 31.4% from geodesy, 8.6% from Europe science and 143% from pedagogy). Almost 36% students, in their opinion, become addicted to nicotine, over 32% students smoked because of relaxing effects of smoking, 129% smoked for company, The biggest group of surveyed group assess their health status as a good (56.3%) and very good (42%), one person as a very bad - 125%. There are statistical significant dependence between health status and gender, age, study, year of study and place of residence. PMID- 19189548 TI - [Motives, awareness of danger and prevalence of tobacco smoking among students of Poznan University of Medical Sciences in Poznan and High School of Country Economy in Kutno]. AB - Tobacco smoking is a common habit both in Poland and around the world--cigarettes are legal and easily accessible. The studied group comprised of 92 students of the Poznan University of Medical Sciences (UM) and 80 students of High School of Domestic Economy in Kutno (WSGK). The aim of the study was to establish causes of tobacco addiction and measure the smokers' awareness of possible health hazards. The studied group took part in an anonymous questionnaire divided into thematic segments, each devoted to one psychoactive substance, including tobacco. The results show that there are more smokers among WSGK students (39% of the surveyed group) than among the MU group (27% of the surveyed students); both universities have a larger population of male smokers than female smokers. The causes for smoking among both groups were quoted as: "for recreational purposes" and "an occasion to socialize". More than 47% of the surveyed UM students and almost 44% of the surveyed WSGK students do realize that tobacco smoking is addictive while 17.4% of the surveyed UM students and 17.5% of the WSGK group understand that smoking poses a health hazard. PMID- 19189550 TI - [The nicotinism question in the light of education on Medical and Nursing Faculties of the Medical University of Bialystok]. AB - Within the framework of "Schedule of health care, economic and social policy pursuing to reduction of tobacco consumption" involving numerous institutions including universities, educated persons will be responsible for the treatment and prophylaxis of tobacco smoking-evoked diseases. The aim of this study was assessment of the tobacco smoking prevalence among undergraduates of last-year course of Medical Faculty and Nursing Faculty of Medical University of Bialystok and evaluation of theirs attitude in the face of the nicotinism question after 5 years of occupation. The percentage of tobacco smokers on Medical Faculty average out 27.7% and 17.8% on Nursing Faculty. Students of both faculties opined that nicotinism in Poland was a heavyweight issue (8 points in decagrade scale). Our data shown that about one third of assessed students have not to do with a nicotinism theme and were not acquired sufficient knowledge to help tobacco smoking patients. Thus, continuation of nictonism issue education within the framework of seminaries and classes of family medicine course in both faculties will be purposeful action. PMID- 19189551 TI - [Analysis of the phenomenon of smoking cigarettes and reasons this phenomenon among the students in the Institute of Cosmetology and Health Care in Bialystok]. AB - The aim of the study was to learn the attitude towards tobacco smoking of students from medical and cosmetic sector. The study was one of the results of the Preventive Medicine Program "Prevention 2006-2009 " implemented at the University of Cosmetology and Health Care in Bialystok. Data was collected through questionnaire which was filled individually and independently by each respondent and additionally anonymity was guaranteed. Research covered group of 501 students in academic year of 2006/2007 and 362 students beginning their studies in 2007/2008 at the University of Cosmetology and Health Care from faculties as: cosmetology, medical rescue, physiotherapy and nursing. The questionnaire covered such topics as: beginning and reasons for cigarettes smoking, the environment of surveyed and attitude of future health service employees towards cigarettes smoking. PMID- 19189552 TI - [Socio-demographic factors and tobacco smoking among the Rzeszow's collage students]. AB - Tobacco smoking is the most known single factor of the highest impact on the number of cancerous deceases. Chemical compound contained in the tobacco smoke have an affect on strong mutagen and carcinogenic actions. Among 400 chemical compound 40 of them have a proven carcinogenic action for human. From the medical point of view the most important are: nicotine- responsible as well as for the pharmacologic tobacco smoking addiction, carbon monoxide, carcinogenic and irritant substances. Regular monitoring of the tobacco smoking among academic students might be used in the actions considering the health risk management. The aim of the study was identifying the relation among selected socio-demographic factors like: age, sex, major, permanent residence, parent's education, material status, medical profile and tobacco smoking among academic students. The research was carried out among 521: science, humanities and medical stationary students. The research was conducted with a usage of the chi-squared evenness test. The research shows that sex and place of residence are the two factors having the most impact on tobacco smoking among academic students. Academic students originated from town or cities as well as men are more often to make a decision on smoking tobacco. PMID- 19189553 TI - [The study of knowledge and attitudes related to smoking habit among students of chosen specializations in Poland and Belarus]. AB - Today, the essentials problem of society is addiction to smoking. The aim of presented study was a comparative analysis of knowledge and attitudes related to smoking habit among students of chosen specializations in Poland and Belarus. All information were collected from the pool questionnaire, which was filled in by each person independently. The questionnaire study was curried out on 806 people: 483 of them study on Yanka Kupala State University of Grodno and 323 of them study in The Institute of Cosmetology and Health Care in Bialystok. They were guaranteed anonymity. The research touched such questions as beginning and reasons of smoking, number of smoked cigarettes, about the environment of the interviewed persons, knowledge about diseases connected with smoking, trials to break smoking cigarettes. PMID- 19189554 TI - [The attitude of the nursing students of Public Higher Medical Professional School in Opole towards the smoking problems in Poland]. AB - Cigarette smoking has been main reason of the Polish society health hazard and one of the most widespread unhealthy element of the human life style. Aim of the study is to evaluate the attitude of the nursing students of Public Higher Medical Professional School in Opole towards the smoking problems in Poland. Most of respondents considered the nicotinism problem in Poland as very important--3 of them evaluate importance of problem on the scale of 0 - 10, estimated it from 8, 9 and 10 points. 74.3% of respondents support the opinion to put the total injunction from smoking at public areas into practice. According to respondents, the most effective forms to express a non-smoking lifestyle is to promote the idea of the total injunction from smoking at public areas and the promotion of the nonsmoking people at the mass media. PMID- 19189555 TI - [The nursing students of Public Higher Medical Professional School in Opole spreading non-smoking lifestyle]. AB - The promotion of health is a science and art of helping people to change their environment and lifestyle to a health friendly one, in order to strengthen and build up their wellbeing. Smoking cigarettes is a most disadvantageous element of a lifestyle. Important elements of promotion of non-smoking lifestyle, by our students, as future Health Service members, are: promoting of smoke-free environment, encouraging of non-smoking fashion, education of health consequences of smoking cigarettes, motivation to quit smoking, advisement in the field of smoking addiction treatment. Public health and health promotion, these are classes where smoking cigarettes' problem has been discussed mostly. Over 90% of students' respondents claim that were able to give a nonsmoking advice to any patient. Because of the awareness of health threats caused by smoking cigarettes and because of the role of education in prevention and addiction fighting, over 82.4% of students were convinced that every doctor should ask every patient, about his/her attitude towards smoking cigarettes. PMID- 19189556 TI - [Cigarette smoking by the mothers of neonates and small children from Lublin area -survey studies]. AB - Cigarette smoking by women in the reproductive age is a threat to the health of a woman as well as a child, in every stage of its life. The investigated group included 189 women aged 19-39. The survey studies were conducted among 123 women staying at the Clinic of Obstetrics and Obstetric Pathology of the Medical University in Lublin as well as 66 women who reported for the treatment at the Department of Conservative Dentistry, having been mothers of at least one child. The questions included in the survey concerned cigarette smoking by the women in the reproductive age considering their dwelling place, education and reporting for their oral health control check-ups. Study results revealed, that from the among 189 women cigarette smoking was reported by 12.70% of the respondents including 20.83% with higher education, 70.83% with secondary education, 8.33% with elementary education. 87.50% smoking women lived in the city whereas 54.17% respondents had one child and 45.83% at least two children. In the group of smoking women 8.33% reported that they limited the number of cigarettes during the day in pregnancy and 29.17% continued cigarette smoking. In the group of nicotine abstainers 56.97% women had higher education, 42.42% secondary education, 0.61% elementary education. 75.76 % lived in the city while 64.24% women had one child, 3576% at least two children. PMID- 19189557 TI - [Cigarette smoking in medical personnel and evaluation of this problem by smoking and non-smoking patients]. AB - Cigarette smoking is a problem in all environments, including health service workers. It increases the number of ill people and accelerates death. The aim of this study was to diagnose the problem of smoking in medical staff and evaluation of this problem by patients. Research was made in hospitals and out-patient clinics in Opole by using an anonimous questionnaire. Along years there has been a decrease of smoking initiation age: 60-years-old-women had their first cigarette in 70% after them finished 18 years old, while most 30-year-old-women had it before. Every year the level of education in medical staff grows up, but the number of smokers in them does not fall down. It is still common to smoke in non-smokers and pregnant woman presence in spite of knowledge about passive smoking. Also pregnancy is not always strong argument to complete quit smoking, among medical staff as well. Smoking medical personnel has definitely negative evaluation by non-smoking patients (70%), a bit less negative it is seen by smoking patients. As the research showed, promotion of nonsmoking workers by employers could be a motivation to quit smoking. PMID- 19189558 TI - [Smoking and passive exposure to tobacco smoke among members of medical rescue teams]. AB - In Poland approximately 33% adults are active smokers. Both active smoking and passive exposition to tobacco smoke leads to serious diseases and causes many other problems. This is why the decrease of smoking prevalence seems to be one of the most important targets of contemporary medicine. In this task the attitude of medical staff plays a great role (prevention, education, behaviour of a medical employee as a positive example). The research was carried out among 48 employees of Regional Station of Ambulance Service in Poznan (16 physicians, 19 medical rescuers and 13 nurses). The group comprised 21 women and 27 men. The investigative tool was a questionnaire concerning smoking, passive exposition to tobacco smoke and attitude of the examined persons towards smoking. We found that smoking prevalence in the investigated group considerably exceeds the average values for Polish population (particularly the number of smoking women) whereas passive exposition to tobacco smoke affects everyone in this group. In the opinion of the examined people the main factors inducing and maintaining tobacco smoking are: addiction, pleasure connected with smoking or influence of someone's company. In a case of medical personnel tobacco smoking (similarly to other behaviours which are harmful for someone's health) weakens the authority of a health care provider as an example of health promotion. PMID- 19189559 TI - [Tobacco smoking among nurses--a lack of knowledge or effect of work overload?]. AB - Nursing profession is characterised by a number of workload modes, both psycho social and purely physical in nature. There are those typically present in all care-related professions as well as those specifically associated with nursing personnel workplace. Such workload categories characteristically result in negative consequences both observed in personal functioning sphere and in wider, social aspect. Nurses constitute, beside medical staff, an essential pillar among medical professions. Full realisation of preventive, therapeutical and rehabilitative functions would be not possible without them. However, as it is frequently being noticed, nurses' professional activity takes place in difficult conditions, resulting from the lack of necessary resources. Nursing personnel are also considerably burdened physically, both throughout preparations to, and during actual care over the patient, and psychically as well, as a result of functioning among ill persons. Medical condition and suffering of the latter substantially contribute to the resultant perception of working conditions by nursing personnel. The present article focuses on examining the relationship between tobacco smoking among nurses, and their perception of being overloaded by work environment requirements. The study was conducted in 2008 throughout the territory of Wielkopolska (Great Poland). It comprised 118 persons employed in nursing subsystem. Diagnostic survey was the employed method, with the use of questionnaire as a research tool. The study revealed that the performance of nurses' professional duties is more often than not accompanied by elements of quantitative work overload, affecting somatic health, as well as those of qualitative overload, which induced undesirable effects in the psychical sphere of wellness. It was also established that 27.12% of the population under study smoked. The addiction frequently happens to be a way to have a pleasant rest after exhaustive work. PMID- 19189560 TI - [Frequency and location of inhaling tobacco smoke by the still smoking participants of the 'Quit and Win' competition who live in cities]. AB - Mount of toxic chemical substances supplied to the human organism is directly proportional to the number of smoked cigarettes. Passive smoking is harmful not only for the non-smokers but also for the smokers themselves. The aim of the work is to establish the number of smoked cigarettes and location where tobacco smoke is inhaled at home, together with checking the differences in behaviour concerning smoking tobacco among the still smoking participants of the 'Quit and Win' competition who live in cities. Leading long-term researches, in August 2006, another questionnaire was sent by post to 1700 participants of the 'Quit and Win' competition organized in 1996 in Poland at the end of the 2nd International Antinicotine Campaign. Answers were sent back in by 648 people 550 of whom (which is 84.9%) lived in cities. Ten years after joining the 'Quit and Win' competition, from among 550 respondents living in cities, 126 (22.9%) said they still smoke. 52.4% (66 people) of those still inhaling tobacco smoke lived in big cities, which is basically Lodz, 23.8% (30 people) lived in cities with fifty to two hundred thousand people and also 23.8% (30 people) lived in cities below fifty thousand inhabitants. The ratio of smoking respondents living in big cities was 27.0% (66 people), in cities with fifty to two hundred thousand people - 19.6% (30 people) and among respondents living in cities below fifty thousand inhabitants, also 19.6% (30 people). People smoking a large number of cigarettes, that is from 11 to 40 cigarettes a day made up for the majority of the respondents living in big cities (44 people, f = 0.67), whereas the lowest number (11 people, f = 0.37) was among the respondents from cities between fifty to two hundred thousand inhabitants. People who could not quit smoking throughout the ten-year period (126 people) were asked in which part of the house they smoke most often after work. The analysis of the answers allowed to draw a conclusion that 72 people (57.2%) smoked mostly in the house, in the room (51 people = 40.5%) or in the bathroom, or the kitchen (21 people = 16.7%), whereas 54 respondents (42.8%) concluded that in order to smoke they usually go out to the hall or to the balcony. Those who smoked at home but outside the flat, were primarily from the cities of below fifty thousand inhabitants (17 people, f = 0.57), whereas the smallest part was made up of respondents living in big cities (24 people, f = 0.36). The obtained results proved that frequency and location of the inhaled tobacco smoke by the still smoking participants of the 'Quit and Win' competition living in cities differed significantly and depended on the size of the city. PMID- 19189561 TI - [Smoking among patients hospitalized in SPSK 4 in Lublin]. AB - Approximately 10 mln of Poles smoke 15-20 cigarettes per day. In total Poles smoke 4,5 billions of packaging each year. Nicotine is not only major threat to humans' health but smoking is also one of the most common elements of any healthy life style. The aim of the work was to analyze factors which influence nicotine addiction as well as to evaluate motivation among patients to stop smoking. The study was conducted among patients hospitalized in the period between January 2006 and December 2007 in SPSK 4 in Lublin. Participation in studies was freewill and anonymous and choice of respondents random. In order to collect necessary information questionnaire was created. First part includes questions as regards age, gender, place of residence, marital status, education and economic situation. Second part includes the right questionnaire which is Fagestrom and Schneider test. Questionnaire was filled by 2346 patients who were chosen randomly. 36.23% of patients admitted smoking. 79.05% of patients are slightly addicted to nicotine, whereas 20.95% are in medium or in higher group of addiction. Analysis of willingness to give up smoking proved that 56% of respondents want to give up smoking. On the base of performer research we can conclude that: patients, who based on the results of Fagestrom's test, qualified into high level addiction are at increased risk of developing nicotine-related diseases and healthy behavior as well as nonsmoking should be promoted among patients. PMID- 19189562 TI - [Tobacco smoking in a sample of middle-size city inhabitants aged 35-55]. AB - Tobacco smoking constitutes a common risk factor for the majority of civilization diseases, such as cardiovascular system diseases, malignant neoplasms and digestion and respiratory system disorders as well. Tobacco-related disorders relate to exacerbation of chronic diseases, for example diabetes and multiple sclerosis. Poland is one of those countries, where the prevalence of smoking is especially widespread. In Poland 42% of men and 25% of women smoke cigarettes and the amount of addicted people amounts to approximately 10 million. The latest data from the year 2003 show that the amount of cigarettes smoked by a particular citizen in Poland has risen fourfold since the beginning of 21st century. This paper presents an analysis of prevalence of tobacco smoking among inhabitants of a middle-size city in the Lodz province aged 35-55 years. The study sample comprised 124 people, including 75 females and 49 males. The tool of the research was a questionnaire survey containing questions concerning cigarette smoking. The study found out that 39.5% of respondents (41.3% of females and 36.7% of males) smoked cigarettes. The percentage of former smokers amounted to 15.3% and the percentage of non-smokers was higher than regular smokers and amounted to 44.8%. The study results showed that the majority of smokers were in the age interval of 45 to 49. Cigarette smoking influenced on smokers' health. The blood pressure and lipid balance was higher among smokers than among people who did not smoke cigarettes. The results of the conducted study confirm that there is a strong need of implementation of programmes towards limiting tobacco smoking, which may contribute to lowering the risk of tobacco-related diseases. PMID- 19189563 TI - [The influence of social-demographic attributes on tobacco smoking by elderly people]. AB - The quitting of tobacco smoking has advantageous influence on morbidity and mortality in each age. A reduction of deaths for cardiovascular reasons in the earliest. The research has taken over a random sample of people aged 65 and more, living in Lodz- Gorna district. The 57.4% turnout to the research enabled to obtain data from 828 people (288 men and 540 women). The respondents were 71.5 +/ 5.1 years old on average. The tool of the research was the interview questionnaire including smoking tobacco related questions. The research was conducted within CINDI WHO Program in 2002. In the big city population the elderly people smoking frequency was 10.8%, while there were 9.5% of regular smokers. Twice as many men than women at the same age were smoking tobacco (16.6% vs. 7.8%). The percentage of heavy smokers was 44.3%. The proportion of regular smokers diminished together with the age and one observed a drop of the average number of cigarettes smoked. One stated that the age and sex are the strong predicators of smoking whereas the high of income and self-appreciation of health state have no influence on smoking by elderly people. The smoking frequency of elderly people living in the city environment is quite high, especially within men 65-75 aged. PMID- 19189564 TI - Health evaluation of the 2nd International "Quit and Win" Antinicotine Campaign participants ten years later. AB - Smoking is one of the most often noticed types of negative behaviour among the Poles. In the work, the results of the health evaluation are presented of the participants of the 'Quit and Win' competition ten years after making a decision to refrain from smoking, also the dependency between this evaluation and behaviour connected with smoking among the people living in big cities and small towns and villages was analysed. Among the 648 respondents, majority, which is 302 people (46.6%) evaluated their health as good, 236 (36.4%) as average, and 76 of the questioned (11.7%) as very good, 29 people (4.5%) as bad, and 5 of the questioned (0.8%) as very bad. The respondents most often evaluated negatively their health in the group of the still smoking living in the big cities, and the least often in the group of the non-smokers living in small towns and villages. PMID- 19189566 TI - [Effectiveness of antismoking counseling for postpartum women]. AB - The aim of the study was to evaluate the effectiveness of antismoking counseling for postpartum women. The study was conducted between 2004-2006 in Lodz district. The study population consisted of 199 women and among them 100 were included into the intervention group and 99 to control one. Antismoking intervention and control activities were conducted among the women who quit smoking during pregnancy and maintained smoking abstinence after delivery. There were no statistically significant differences regarding the socio-demographic characteristic between intervention and control group. The women who received antismoking advice relapsed into the smoking less frequently than the women from the control group (26% vs. 56%; OR = 0.25; 95% CI 0.15-0.51). The antismoking counseling for postpartum women is the effective tool to help them to maintain smoking abstinence. PMID- 19189567 TI - [Predictors of smoking cessation in group-based behavioral intervention programme -research findings in 2001-2007]. AB - The efficacy and cost-effectiveness of behavioral treatments compare favorably with the pharmacotherapies and community-based interventions. Group-based behavioral programmes have been scientifically proven as the effective smoking cessation intervention. Identifying predictors of the efficacy of smoking cessation within group-based programme using characteristics of participants (social and environmental approach). Program is a multicomponent group-based behavioral intervention with the elements recommended by the US Public Health Service as the most effective. 517 smokers were included into the program in the outpatient clinic setting in years 2001-2007. A point prevalence abstinence (PPA) was estimated by self-reported smoking cessation. 59.% of participants stopped smoking during four-week program. Using multivariate logistic regression, a model provided the optimal predictive ability. PPA decreased (odds ratio [OR], 0.7; 95% confidence interval [CI], 0.24-0.93) for manual occupied and unemployed participant comparing to nonmanual occupied smoker, decreased (OR, 0.9; 95% CI, 0.20-1.19) for high-nicotine-dependent smoker, increased (OR, 1.49; 95% CI, 0.74 2.98) for high-motivated to quit smoking participant, increased (OR 2.31; 95% CI, 1.15-4.66) for subject smoked 1 pack or more of cigarettes per day, decreased (OR 1.41; 95% CI, 0.85-2.35) for participant with smoker in household, decreased (OR 1.74; 95% CI, 1.09-2.78) for smoker without any previous physician's advice to quit and did not distinguish for subject without any previous attempts. The future of behavioral group-based treatment will likely emphasize intensive intervention targeted to particular subgroups of smokers with the need and motivation for them (eg, the low nicotine-dependent but with high emotional and/or behavioral components of dependency, poor supported by family and non medical patients). A blurring of the distinctions between behavioral interventions, pharmacotherapies, and community-oriented approaches is also likely as multidimensional cessation strategies are developed. PMID- 19189568 TI - [City dwellers' behaviour concerning smoking ten years after joining 'Quit and Win' competition]. AB - Harmful influence of chemical compounds on human organism included in tobacco has been thoroughly tested and described in literature. The results of tests show quite clearly that it is worth undertaking actions that aim at limiting and perhaps even eliminating the habit of smoking in society. Antinicotine actions led in big populations are considered an effective and cheap method of fighting the habit of smoking. The aim of the work is to present the differences in behaviour as far as smoking is concerned, among the inhabitants of cities and towns, depending on the place's size, ten years after joining 'Quit and Win' competition. In August 2006 there were additional tests o 1700 participants of 'Quit and Win' competition that ended in 1996 and finalized the 2nd International Antinicotine Campaign in Poland. A survey sent by post was used. Answers were sent back by 648 respondents, 550 of whom (which is 84.9%) lived in cities. People declaring total tobacco abstinence underwent biochemical tests. Ten years after the attempt to sustain tobacco abstinence, since May 1996, 424 people living in cities came to the conclusion that they achieved their goal and do not smoke at all, whereas 126 people (22.9%) could not reach the total abstinence throughout such a long period of time. The non-smokers were 77.1% of all the respondents and at least 34.0% of all the 'Quit and Win' competition participants who lived in cities. 178 people living in cities claiming they did not smoke at all made up for 73.0% of the respondents and at least 26.5% of all the competition participants living in the cities. In the cities with the number of inhabitants numbering from fifty to two hundred thousand, 123 who were already non-smokers made up for 80.4% of the respondents an at least 36.4% of all the participants of the competition living in the cities with such a number of people. In the cities where there were less than fifty thousand people, 123 non smoker also made up for 80.4% of the respondents and at least 51.9% of all the competition participants from the cities with less than fifty thousand inhabitants. The results prove great efficiency of the 2nd International Antinicotine Campaign in Poland that ended the 'Quit and Win' competition. The analysis of the collected material shown that after ten years, participation of the nonsmokers from the 'Quit and Win' competition statistically depended very much on the size of the city. The difference between the size of the ratio of non smoking participants living in big cities (Lodz) and the ratio of non-smoking participants living in the cities of fifty to two hundred thousand inhabitants was 9.9 percentage point, whereas between the ratio of non-smokers living in small cities of less than fifty thousand inhabitants was as much as 25.4 percentage point--to the benefit of the small city dwellers. PMID- 19189569 TI - [Opinion of students of the Medical Department of Wroclaw Medical University on the subject of the fight against tobacco addiction]. AB - Problem of prophylaxis of nicotine-related diseases and leading of programs of the fight against tobacco addiction are being given out to be shared nominative case of main plans led in the framework of health care in majority of European countries. An anonymous questionnaire containing questions concerning students' opinion on the subject of the fight against tobacco addiction was carried amongst students of the 6th year of the Medical Department of Wroclaw Medical University in the academic year 2007/2008. 177 students took part in the study (64% of women, 34% of men). 72.4% of students came from cities with the population above 100 hundred of inhabitants. 64.4% of students is regarding problem of nicotinism in Poland as important. 62.2% of students think that promoting not-smoking in media is most effective method. 76.7% of students is justifying total smoking ban in public places and is behind accepting such regulations in Poland. 1.14% of students is against smoking ban in public places regarding it as limiting their freedom. Majority of students thinks that nicotinism is an essential problem in Poland. Dominating percentage examined thinks that promoting not-smoking in media is most effective method. Majority of students is justifying a total smoking ban in public places and is behind accepting such regulations in Poland. Only scarce percentage examined is against a smoking ban in public places regarding it as limiting their freedom. Undoubtedly students notice problem of tobacco smoking in Poland as important, at the same time rarely taking into consideration keeping anti-tobacco programs on the level of health care--are handing responsibility for realization of these programs to media or government organizations. Students are supporting regulations introducing a ban on smoking in public places. PMID- 19189570 TI - [The use of the minimal intervention strategy among police officers]. AB - It has been estimated that current tobacco smoking will cause about 450 million deaths worldwide in the next fifty years. The intense search for effective methods of smoking reduction continues. Minimal Intervention Strategy (MIS) is a simple, cost saving as well as efficient method of smoking limitation. The survey analyzed the use of MIS by physicians among policemen from the economically active population. The tool of the research was the interview questionnaire including 49 smoking tobacco related questions. The research encompassed the group of 102 policemen from the headquarter in Tomaszow Mazowiecki. The respondents were 36.0 +/- 7.36 years old on average. The smoking frequency of the surveyed group was 66.7%, where there were 56.9% of regular smokers and 9.8% of occasional smokers. The percentage of never smokers reached the level of 28.4%. Physicians used the first step of MIS (question about smoking) in case of 55.9% respondents. However, only 1/3 of regular smokers were recommended to stop smoking (the second step of MIS). CONCLUSIONS: The tobacco smoking frequency among police officers was very high. The use of MIS in the surveyed group should be considered insufficient. PMID- 19189571 TI - [Realization of problems related with tobacco addiction under medical studies]. AB - Quitting by current smokers is therefore the only way in which tobacco-related mortality can be reduced in the medium term. An anonymous questionnaire containing questioners on the subject of the realization of problems connected with the tobacco addiction in the route of medical studies was carried amongst students of the 6th year of the Medical Department of Wroclaw Medical University in the academic year 2007/2008. 177 students took part in the study (64% of women, 34% of men). 72.4% of students came from cities with the population above 100 hundred of inhabitants. 8.5% of students isn't regarding the nicotine addiction as illness. 94.4% of students is claiming that problems concerning nicotinism were being brought up on the university. 81.7% of students is judging that he is able to give an anti-smoking advice to a patient. 89.8% of students think that every doctor should ask each patients opinion about smoking cigarettes independently of whether he is an ill or healthy person. It is surprising that persons not regarding nicotine addiction as illness are still amongst students of 6th year of the Medical Department. Not all examined also confirmed that they had met with discussing the problem of nicotinism during medical studies. Majority examined thinks that he is able to give an anti-smoking advice to a patient. Dominating percentage examined thinks that every doctor should ask each of patients' opinion about smoking cigarettes independently of whether he is an ill or healthy person. Knowledge is being passed down to students on the subject of nicotine addiction during main subjects realized during medical studies. However this knowledge seems insufficient in spite of high self-assessment of students. PMID- 19189572 TI - [Tobacco smoking is addictive--do not start smoking]. AB - Tobacco smoking is addictive- do not start smoking, do not start smoking, each person buying a package of cigarettes can read this inscription. But does he really read it? Every year all over the world one million people try to stop smoking, but only for a few percent this attempt is successful. Giving up the habit of smoking is hard because it leads to biological and psychogenic addiction. The aim of this paper was to check which factors most motivated smokers to stop smoking? 50 active smokers, among them 5 former smokers, who have smoked at least 15 cigarettes a day for 15 years were classified to the preliminary study. Among them there were 23 women, which is 46% of the subjects and 27 men, which is 54% of the subjects. The average number of smoked cigarettes among present smokers was 18.4 pieces, and the former smokers smoked the average of 19 pieces daily. Present smokers smoke 8 years longer, i.e. 26.3 years than former smokers. Inscription placed on cigarette packages concerning the loss of health definitely influence smokers to make another attempt to stop smoking. This important concerns the appearance of lung cancer, the differences were statistically significant between former and present smokers (p < 0.03), women are afraid of lung cancer twice more often than men (p<0.02), also women more often were afraid of cardiovascular system diseases (chi2 = 2.013, p < 0.03) and painful death (chi2 = 7.729, p < 0.006). Over 80% of smokers declared that the raise of the price of cigarette package has a significant influence on further attempt to stop smoking. Among the subjects 20% declared that ad spots on TV positively influence giving up smoking. Other factors such as: chewing gum (only 11.1% willing to try again), tablets (only 8.9%), appearing cough (2.2%), unpleasant tobacco smell (2.2%), the loss of taste (0.0%), the improvement of life comfort (4.4%) at minimum extent motivated smokers to stop smoking. Unsuccessful attempt of giving up smoking should not discourage any smoker no his doctor to further fight for his health. In another attempt the arguments that most motivate the smoker to stop smoking are worth using again. PMID- 19189573 TI - ["How does smoker really smoke?"--preliminary report on smoking topography among Polish smokers]. AB - Levels of toxic substances in tobacco smoke are undeniably influenced by temperature-oxygen conditions in which the smoke is generated. These conditions depend on the way the cigarette is smoked (smoking topography). Smoking topography may be characterized by such factors as: puff volume and its velocity, intervals between puffs and a number of puffs per cigarette. Vast majority of formerly published papers on tobacco smoke composition present data obtained in accordance with ISO (International Standard Organization) and FTC (Federal Trade Commission) standards concerning conditions of tobacco smoke production. Currently, the standard methods for tobacco smoke generation in lab conditions are frequently questioned by researchers, since such methods do not give reliable results as far as toxic substances delivery to smokers' bodies is concerned. To determine the real doses a tobacco smoker is exposed to, first of all smoking topography should be measured in a given population. The aim of this study was to characterize smoking topography among Poles and its statistical assessment. 129 volunteers were involved in the research. To assure representativeness of the group of smokers, in the first step of the study a demographic structure analysis of smokers' population in Poland was carried out (the authors used data provided by GUS (Central Statistical Office). Smokers were divided into study groups in terms of their sex and age and also detailed information on tobacco addiction (number of cigarettes smoked, its brand and type etc.). Smoking topography was measured using a portable CreSSmicro device (Plowshare Technologies, USA). Mean puff volume in the examined group was 60 ml and was 78% higher than the puff volume used for tobacco smoke control according to the ISO method (35 ml). Mean measured puff velocity (48 ml/ sec.) was as much as 120% higher comparing to ISO (17.5 ml/sec.). Mean duration time of puff in the examined group was 1.7 sec. (2 sec. in the ISO standard method) so the difference was not very significant but still 13.2%. However, mean interval between puffs in our group was 20 sec.- threefold lower than in the ISO method (60 sec.). Moreover, examining the variability of subsequent puffs, the authors found out that the puff volume lowers as the cigarette is smoked. All measured smoking topography parameters were highly dispersed within the examined population of smokers. PMID- 19189574 TI - [Health behaviors of workers in context of act of smoking permission in 2006-2007 yrs]. AB - Smoking defines as a gate open for development other addictions in a group of adolescents and educational institutions have significant influence on development of health behaviors. The aim of the study was the estimation the phenomena of respecting of law in area of smoking permission in Poland in 2006 2007 yrs. Schools and other educational institutions were considered especially. Over 209 000 work place participated in the observation and 46 000 of educational institutions were in that group. Permission of smoking cigarettes was respected in 99.3% of educational institutions. 15.1% of bars and restaurants respected the law in that area. The biggest problems in respecting the law were observed in Gorzow Wielkopolski and Bialystok. The results show there is the necessity of leading more efficient educational actions connected to health behaviors at schools and other institutions and finding new solutions for respecting the law. PMID- 19189575 TI - [Legal regulations knowledge among active and passive smokers]. AB - In the recent years smoking problem is more visible in Europe as well as in Poland. European countries tried to limit smoking in different ways including suitable legal regulations. There came international legal documents. Besides of that some countries implemented solutions to protect passive smokers. Also in Poland there are legal acts protecting non-smokers from passive smoking. Those provide protection from nicotine smog in sufficient way but those are often ignored. More effort should be put in the implementation of existing acts. The main research objective of this document was to show the actual legal regulations knowledge about smoking and selling tobacco by active as passive smokers and their opinion on smoking prohibition in public places. PMID- 19189576 TI - [Binary hypothesis testing with ROC curves]. AB - In medical research we frequently need to delimit some areas of the measured parameters to use them in diagnosing existence of some kind of abnormality. In the article the usage of ROC curves and comparison of obtained results to the outcomes of logistic analysis, decision trees and reference intervals is described. PMID- 19189577 TI - [Genes associated with tobacco smoke-associated cancer of head and neck]. AB - The article presents the current techniques used for the identification of genes involved in tobacco smoke-associated cancers. The focus is set on the techniques derived from the conventional cytogenetics and includes fluorescence in situ hybridization (FISH), comparative genomes hybridization (CGH) and its further improvement that is array-CGH, and other aspects of microarray DNA technology. The second part deals with the main findings concerning participation of oncogenes and tumor suppressor genes in development and progression of tobacco smoke-associated head and neck cancers. PMID- 19189578 TI - [Screening studies--is tobacco smoking always recognized as a risk factor?]. AB - There were introduced several screening programs for the early detection of neoplasms. They were devoted for diagnosis of cancer at early stage which gives a greater chance for totall recovery from the disease. The screening protocols include proper taking patient's history and then phisical examination with possibility of taking samples for histological evaluation. In this paper the authors on the example of screening of gynecological cytolotogy would like discuss the problem of lack of proper patients' history especially according the smoking habit. PMID- 19189579 TI - [Smoking cessation as regards anesthesia and surgery]. AB - Smoking cessation in the perioperative period allows a substantial reduction of a number of anesthetic and surgical complications. They primarily include pathology and adverse reactions of respiratory, cardiovascular and nervous systems and also alterations in wound and bone healing. There is no doubt that long-term (several months) tobacco abstinence reduces the complication risk; however, the significance of short-term smoking cessation is being discussed. Additionally, due to increased patient motivation surgical operation poses a "teachable moment" to be captured for quitting permanently. Interventions for preoperative smoking cessation effectively change smoking behavior and should be widely offered to surgical patients. Oral recommendations should possibly be supported by pharmacotherapy. Nicotine replacement therapy and bupropion proved to be effective and safe in the perioperative period. PMID- 19189580 TI - [Is tobacco smoking an effective means of body weight control--review of literature]. AB - A lot of people, particularly young people begin smoking tobacco with the belief, that it is an effective means of body weight control. The decision to quit coming in a later stage of life is accompanied by the fear of gaining weight after stopping smoking. The mechanism through which nicotine may be an agent in the process of controlling body weight consists in increased energy expenditure of the organism and reduced appetite. However tobacco smoking is not an effective means of combating excessive weight. This is because it stimulates formation of abdominal adipose tissue, thus increasing the risk of type 2 diabetes and heart diseases. Therapy of tobacco addiction should be combined with diet counselling and promotion of physical activity, which will help to limit the increase of body weight. PMID- 19189581 TI - [Use of hair nicotine as a tool to assess tobacco smoke exposure]. AB - The first paper concerning hair nicotine determination was published in 1983 by Ishiyama et al. Since then, examining hair nicotine content has become a valuable tool facilitating assessment of exposure to tobacco smoke in various groups of people (active and passive smokers or even neonates and fetus). The present article describes briefly recent research studies on the use of hair nicotine as a biomarker. Furthermore, the paper describes various determination techniques related to nicotine in hair, as well as advantages and disadvantages of hair nicotine as a biomarker. PMID- 19189582 TI - [Pharmacotherapy of smoking cessation with application of nicotine and nicotine free drugs]. AB - The article summarise the current knowledge about medicaments used in smoking cessation. In detail are discussed drugs, which are the first choice in treatment of nicotine dependence. The basic preparations of Nicotine Replacement Therapy (NRT) are: chewing gum with nicotine which allowed for instantaneous reaction on craving for nicotine, and nicotine patches which supplement organism with this alkaloid with constant rate. The efficacy of other NRT preparations (vapour inhaler, nasal spray, sublingual tablet, lozenge) are on the some level as nicotine chewing gums and nicotine patches. Alternatives for NRT are two drugs bupropion and varenicline. Both drugs are effective in treatment the nicotine dependence and as well as nicotine containing medicaments can be us as a first line drugs. The efficacy of other drugs (nortryptyline, mecalamine, clonidine) have not been proved in randomized controlled studies. PMID- 19189583 TI - [Promissing role of probiotics in prevention of smoking-related diseases]. AB - Humans are highly adapted to consuming probiotics. Trehalose ("mushroom" sugar) is probably an important reserve compound and stress-responsible metabolite (increases bile and gastric acid resistance) of probiotic bacteria's and trehalase activity, in contrast to lactase activity, is preserved in all human populations, even those not consuming mushrooms. Among traditional Melanesian horticulturists, of whom 4/5 are daily smokers, the diet is rich in pre- and probiotics and there is absence of cancer and cardiovascular disease. Indoor air pollution is an important cancer risk factor. Over thousands of years, the controlled use of fire for preparing meals has resulted in exposure to smoke pollution (biomass fuels produce up to 100 times more respirable particles compared to gas ovens). Simultaneously, up until the 20th century, the only commonly available and inexpensive way of preserving food was fermentation and drying. Probiotics may protect the detoxification function of the kidney and liver. Furthermore, it can be speculated that probiotics may help in adaptation to smoke pollution generated during cooking, heating but also tobacco smoking. Smoking is the most important lifestyle risk factor for bladder cancer and the consumption of probiotic foods reduces the risk of this cancer in humans. Probiotics may restore natural killer cell activity which is lowered in smokers. In one study it was observed that a diet supplemented with Lactobacillus plantarum could be also useful in the prevention of cardiovascular disease in smokers. There is no sufficient data from clinical trials to recommend the routine use of probiotics in prevention of smoking-related diseases. More research is needed to investigate the role of probiotics in this area. PMID- 19189585 TI - [The effect of environmental tobacco smoke during pregnancy on psychomotor child development]. AB - The association between maternal smoking and retarded maternal condition and birth outcome is well know. Smoking during pregnancy increase risk of preterm delivery, hipotrofia, delayed developmental of respiratory and nervous system. The risk for most of thisconditions has been found to increase with the number of cigarettes smoked. The last examination shows that environmental tobacco smoke during pregnancy leads to physical problems, behavioral problems even mental retardation. PMID- 19189584 TI - [Cigarette smoking--threat from first days of life]. AB - It is obvious that smoking is harmful. It is especially dramatic that it may cause a lot of dysfunctions among newborns who without their free will are expose to toxic tobacco components. The consequences are as serious when mother smoke as when she is exposed to environmental tobacco smoke (ETS). Exposure to cigarette smoke during pregnancy is directly correlated to premature rupture of the membranes, premature birth, intrauterine growth retardatyion--IURG, low birth weight, smaller body length of newborn, reduced lung function--hypoxia, delayed development of respiratory and nervous system, sudden infant death syndrome- SIDS, obesity, allergy, arterial hypertension or even tumors. That is why the prophylactic of above illnesses is important since first days of life. PMID- 19189587 TI - [Preparation of a programme entitled "The role of pharmacists in providing support for nicotine replacement therapy in public pharmacies]. AB - In the last years, the role of pharmacist in the community has changed considerably. Pharmacists became active patients' consultants and health confidants. Smoking is one of the most important risk factors for burden of disease and a major contributor to the death of millions of people each year. Pharmacists as competent health care providers should fight against tobacco addiction so their participation in the care of patients who wish to give up smoking would be beneficial. The Chair of Toxicology of the Jagiellonian University established and launched an outpatient smoking-cessation programme based on the public pharmacies. The role of pharmacists in health promotion programmes, including the fight against tobacco addiction in Poland and worldwide was described. We also defined the range of activities which can be undertaken by the pharmacists in order to effectively support patients who want to stop smoking. In Methods we presented the programme preparation phases and the contents of posters and leaflets which promote smoking cessation. We also watched the initial efforts and engagement of pharmacists implementing this programme in public pharmacies. PMID- 19189586 TI - [Novel risk factors for cardiovascular disease and tobacco smoke]. AB - Tobacco smoking, counted as one of the alternating external cardiovascular risk factors, can cause disadvantageous changes of concentration of compounds considered to be the new internal risk factors. The aim of the study was to determine the influence of tobacco smoke on concentration levels of homocysteine, dimethylarginine (ADMA), fibrinogen and C-reactive protein (CRP) in healthy smoking male's blood plasma. 71 healthy male volunteers, aged 30-59, were chosen for the study. Based on a questionnaire they were divided into non-smoking group (37 individuals) and active smokers group (34 individuals). The results obtained indicate the most significant increasing effect of tobacco smoke on blood plasma homocysteine levels (by 11.9%) and then, successively, on fibrinogen (by 10.1%) and ADMA (by 9.1%). Changes of CRP concentration, although differentiated similarly to other factors, were insignificant. Since the coexistence of elevated risk factors shows a synergistic effect on cardiovascular risk level in general, it is advisable to perform tests of new risk factors among people exposed to tobacco smoke. The tests will appoint the ones at risk of disease and help to cover them with a medical care. PMID- 19189588 TI - [Present knowledge on the effects of smoking tobacco on the eye diseases]. AB - It is accepted that tobacco smoking impairs different functions of the organ of vision and can be the cause of different eye diseases. It is well-known that tobacco smoke contains more than 4,000 substances, some of them exerting toxic influence on the eye by producing the ischemia or/and oxidative stress. In previous years, the association between tobacco smoking and cataract, age-related macular degeneration (ARMD), thyroid eye disease, anterior ischemic optic neuropathy and primary open angle glaucoma (POAG) were observed. It was also suggested that smoking might have a negative impact on the treatment of scleritis, thyroid eye disease and surgery of POAG. In last two years many interesting studies have been conducted on the influence of smoking on morbidity and the character of AMD, cataract, vascular diseases of the retina, cystoid macular edema complicating intermediate uveitis, refractive errors, thyroid eye disease and the function of the optic nerve and retina measured by electrophysiological techniques. Concluding there is increasing evidence that tobacco smoking is a risk factor in case of many eye diseases. This suggests that by eliminating smoking, it might be possible to decrease the morbidity of eye diseases in the future. PMID- 19189589 TI - [Evarts A. Graham (1883-1957)--biography and medical achievements. His input into the knowledge of carcinogenic actions of tobacco]. AB - Evarts Ambrose Graham (1883-1957) was one of the most prominent American surgeons of the 20th century. Some believe that he belonged, together with William Halsted and Harvey Cushing, to the three most respected American surgeons. Graham was mainly dedicated to thoraco-surgery. He proposed a new treatment for empyema of the chest, based on the physiological understanding of pleural mechanics. He introduced the cholecystography method for the visualization of gall bladder and performed the first total one-stage pneumonectomy. Moreover, he was actively engaged in the studies of carcinogenic influence of tobacco and an association between tobacco smoking and lung carcinoma. PMID- 19189590 TI - [Effect of tobacco smoking on skin and mucosa appearance, ageing and pathological conditions]. AB - Tobacco smoking is considered to be one of the main factors that negatively affect the skin, mucous membranes and teeth, i.e. the body parts that decide on the so called 'healthy look' and 'attraction'. This paper is a review of literature on the subject of harmful and pathological influence of tobacco smoking on the skin and the immunological system. Furthermore, the paper described such smoking results as wrinkle development, skin elasticity and thickness loss, as well as a decreased ability of tissues to regenerate. Finally, the authors discussed various diseases connected with tobacco use: changes in the skin, teeth and mucous membranes, and cancerous lesions. PMID- 19189591 TI - [Health promotion in a workplace in The United Kingdom of Great Britain and Northern Ireland by taking into consideration acts prohibited tobacco smoking]. AB - This study presents health promotion strategies in a workplace in The United Kingdom of Great Britain and Northern Ireland. A workplace is a very important area for health promotion activities because of almost unlimited access to target group--adult group, especially good opportunity to access to men, who rather seldom going to a doctor. In 2004 Committee on Tobacco and Heath (SCOTH) classified environmental tobacco smoking (ETS) for category: "serious hazards for public health" after conducted study of harmful effects of ETS. In 2007 England becomes "free" from ETS, because of acts prohibited tobacco smoking in public places and in workplaces. In 26th of March 2006 Health Act in Scotland and in the second of April 2007 in Wales also prohibited smoking in public places. The aim of a new act was to protect health and prevention illness caused by exposition to ETS. So, there is prohibition for everyone: employees, clients, employers and guests in a whole company, in closed area and fundamental closed areas (according act). The act is also about previous areas intended for tobacco smoking. In 30th of April in 2007 Northern Ireland also prohibited smoking in public places. There are many profits for everyone, for example: protection people's health exposed to ETS, much slower development some serious illness, reduction of medical care cost and improvement of environment for community. If smokers decide to quit smoking or even reduce the number of smoking cigarettes per day, a population would be healthier. PMID- 19189592 TI - [Tobacco smoking among population in the United Kingdom of Great Britain and Northern Ireland between years: 1950 and 2003]. AB - In the last year there are many strategies focused on a new challenges such as: obesity, tobacco smoking, abuse alcohol, sexual illness in public health policy. There are differences in the prevalence of tobacco smoking because of many factors, for example, social class and education (if there is low social status and education, the prevalence of tobacco smoking in population is higher). General, in Scotland there are much more smokers than in England, Wales and Northern Ireland. The Health Development Agency (HDA) studied that tobacco smoking in 1998 and 2000 years in England caused 9 deaths per 10 men and 8 deaths per 10 women suffered from lung cancer. This study presents prevalence of tobacco smoking among a population of The United Kingdom of Great Britain and Northern Ireland. A paper describes also events influencing the prevalence of tobacco smoking and strategies taken for reducing tobacco smoking among population by British government. PMID- 19189593 TI - [Development of the schizophrenia concept and diagnostic tools for its assessment]. AB - Despite the fact that along the last 100 years a number of diagnostic systems and clinical models of schizophrenia have been proposed, defining this psychiatric disorder in a manner acceptable in both clinical and research settings remains one of the most difficult tasks in psychiatry. While the description of symptoms and signs of the disorder has remained mainly unchanged over the years, the way in which the authors have articulated the varied phenomenological manifestations has been different across periods. Great strides that have been made in recent years have brought about a convergence on a definition of the illness as well as methods of assessment that are acceptable to workers from research centres across the world. The structured interviews brought an organizational focus to the traditional psychiatric interview that provided a strict format and directions to the interviewer, which enables systematic assessment of all the criteria necessary for a diagnosis according to the given diagnostic system. Having a consensually shared set of diagnostic criteria as well as structured assessment devices helped to ensure a more common unit of analysis in research across the world. Though most workers would hail the operationalization of diagnostic criteria as an advance in psychiatry, there remain concerns about the impact that the quest for increased diagnostic reliability might have on validity. Given that the ultimate goal of any diagnostic system is to provide insights into the aetiology, pathophysiology and treatment of the disorder, examining various alternative diagnostic constructs and their validity is still an important area of schizophrenia research. PMID- 19189594 TI - [A questionnaire of insight into schizophrenia--"my thoughts and feelings"]. AB - AIM: Construction of a brief, self-rating questionnaire of insight into schizophrenia, that can be useful in everyday practice. METHOD: The questionnaire is based on reports of patients with good remissions, confronted with the group interviews with severely psychotic patients without insight. They led to the formulation of 16 questions with three possible answers reflecting lack of insight, full insight and partial insight. 6 items that have the most discriminating power of patients with and without insight were selected in the study of 367 patients assessed by psychiatrists acting as the competent judges. The reliability of this questionnaire is good--alfa Cronbach = 0.8. In the next study with 128 patients, a similar reliability was fund--alfa Cronbach = 0.77. The validity measured as the correlation with PANSS item "Lack of Judgment and Insight" was acceptable r = -0.55. RESULTS: The 6-items questionnaire of insight into schizophrenia--"My thoughts and feelings" has good reliability and acceptable validity. The discussion of results with the patient, based on rules of cognitive behavioural therapy aiming at the shaping of insight was described as well as the preliminary sten norms were described. CONCLUSIONS: The questionnaire may be useful in the screening of patients with low insight into schizophrenia as well for a psychoeducative discussion of its results. PMID- 19189595 TI - [Brief measure to assess perception of self-influence on the disease course. Version for schizophrenia]. AB - AIM: Elaboration of a brief scale for screening of perception of self-influence on the course of schizophrenia, which could be used during usual psychiatric examination. METHODS: Group focus interview of patients with favourable and unfavourable course of schizophrenia led do the formulation of the 57 items Likert's scale. The study of 403 persons with schizophrenia, assessed by their psychiatrists as having low or high perception of self-influence on the course of schizophrenia resulted in a choice of 5 items that had the strongest power of discrimination of those two groups of patients. The psychometric proprieties were assessed in the group of 90 patients with schizophrenia. RESULTS: The reliability Alfa Cronbach = 0.78 and validity (r = 0.45) assessed by correlations with the clinical assessment CONCLUSIONS: Brief measure to assess perception of self influence on the disease course--Version for schizophrenia has good reliability, acceptable validity and can be used for screening of patients with low perception of self-influence on the disease course. PMID- 19189596 TI - [Needs of persons with mental disorders--definitions and literature review]. AB - Failure of the deinstitutionalization process and a tendency of limitating health care costs on the basis of evidence based medicine initiated planning mental health services according to the clients' needs. Defining needs is difficult due to the complexity of the concept ofneeds, so numerous definitions of needs have been proposed. In the health care context a need is most often regarded as "the lack of health/wellbeing or the lack of access to care". Many clinical and population-based studies on needs were conducted in England and other well developed countries. According to their results the most important are unmet needs, which correlate with unemployment, single status, low quality of life, low satisfaction with services and high social disability scores. Assessment of needs becomes a part of routine clinical practice and evaluation of mental health services. Results of such an assessment enables to fit the therapy and rehabilitation to the individual patient's needs which in consequence leads to a better therapeutic response and lower social disability. First Polish studies on needs of persons with different mental disorders indicate the social needs (company, intimate relationship and sexual expression) as the most often unmet from patient's point of view. PMID- 19189597 TI - [CANSAS--Camberwell Assessment of Need Short Appraisal Schedule and other needs of persons with mental disorders assessment tools]. AB - According to the new tendency of planning and developing mental health services in response to the individual needs, many tools assessing needs of persons with mental disorders were created during last few years. This paper presents some of them, with concentration on the Camberwell Assessment of Need Short Appraisal Schedule (CANSAS). The aim of CANSAS is to identify patient's needs, thus the tool can be a part of routine clinical practice and research, as well as a component of service evaluation. Because "need" is a subjective concept, CANSAS incorporates separately the views of both users and staff about needs. The tool was developed in the Institute of Psychiatry King's College in London and is now successfully used in many countries all over the world. It has been also translated into Polish and used in the first Polish studies on the needs of persons with mental disorders. PMID- 19189598 TI - [Self-concept in schizophrenics--a survey of studies]. AB - The article is a survey of studies concerned with self-concept, self-esteem and self-acceptance in schizophrenics.The aim of the paper is to present current research on self-image (the term 'self-concept' has been increasingly used in recent studies) although without critical analysis. In specialist literature one can distinguish three basic trends of research concerned with the relationships between the self-concept in schizophrenics and the illness process. The first one relates to the nature of self-concept in schizophrenics and contrasts it with that of normal control subjects, as well as includes the research on self-esteem and self-acceptance levels. The other trend takes into account the sex of the examined patients and the third focuses on interrelations between self-concept and other illness-related factors. The article focuses on the first and the second trend. It comprises of three parts. The first one presents the current research on self-concept, self-esteem and self-acceptance in schizophrenics. The second part looks into the relationships between the self-concept and other factors (e.g. disease duration, psychopathological symptoms, therapy). The last part is an overview of research on self-concept and perception of others, as well as the perception of patients by others. PMID- 19189599 TI - [Personality changes in paranoid schizophrenics in perception of patients and their parents]. AB - AIM: The aim of the paper was to describe and compare personality changes in paranoid schizophrenics in the perception of patients and their parents. METHOD: 50 paranoid schizophrenics (ICD-10) and their parents were tested using the ACL test. Patients completed the ACL test using instructions "I am" and "Before the illness I was", while for the parents it was: "My child is" and "Before the illness my child was." RESULTS: Differences between real and retrospective images were treated as the measure of changes. There were 10 differences in the perception of patients, 23 differences in perception of the mothers and 28 differences in perception of the fathers. All these reached statistical significance. CONCLUSIONS: The sense of self-change in patients is seen as a lower need for exhibition, dominance, aggression, autonomy, self-confidence, in personal adjustment and ideal self scale while higher need for abasement, deference and self-control. Mothers notice more than twice and fathers almost three times more the changes in personality of their children than their children themselves. Lower need for exhibition, dominance, self-confidence, personal adjustment and in ideal self scale and higher need for abasement was observed in the group of patients as well as in the group of mothers and fathers. The level of perception of personality changes in children was similar for both mothers and fathers. PMID- 19189600 TI - [Compliance in the treatment of schizophrenia--project ADHES (the renewed results)]. AB - AIM: The aim of this study was to analyse the level of compliance measured by patients, their families and psychiatrists. METHOD: 2009 patients who suffered from schizophrenia, 1791 members of their families and 203 psychiatrists were studied. They answered a questionnaire about the level of compliance and the factors which influence it. RESULTS: The level of compliance in schizophrenia was evaluated by psychiatrists as 48%, families--73% and patients--85%. The psychiatrists assessed that 60% of the schizophrenic patients did not have the sense of illness. 11% of the patients said that they have no benefits from pharmacotherapy. CONCLUSIONS: The level of compliance is the least in doctor's evaluation. The patients evaluation of the level of compliance was the biggest. The level of compliance was related to patients' insight, negative attitude to pharmacotherapy, cognitive deficits. Women were more cooperative with medication than men. PMID- 19189601 TI - [Comparison between the day-care ward and the inpatient ward in terms of treatment effectiveness based on the analysis of psychopathologic symptoms, subjective quality of life and number of rehospitalisations after discharge]. AB - AIM: To assess effectiveness of treatment in the day-care and inpatient wards within the Wroclaw branch of the international multicenter EU-funded EDEN study. The authors analysed the number of rehospitalisations after discharge from the index hospitalisation as well as psychopathologic symptoms and subjective quality of life. METHODS: Out of 1089 patients admitted to the mental hospital in Wroclaw, 238 were randomly assigned to either the day-care ward (n = 115) or the inpatient ward (n = 123). There were three interviews with patients: upon discharge, three and twelve months later. Scores of the extended version of Ventura's BPRS were used for representing the psychopathology level and for calculation of between-setting differences. Subjective quality of life was measured using the MANSA questionnaire. Statistical parameters of effectiveness (scores at given time-point, models adjusted for the scores at admission and for the duration of hospitalisation) were analysed. The frequency of rehospitalisations was also investigated. Statistical analyses after discharge were performed using the random multiple imputations method in consecutive time points. RESULTS: There were no statistical differences between settings in the effectiveness of treatment three months after discharge. Interviews performed one year after discharged revealed a higher level of psychopathology in patients treated in an inpatient setting (p = 0.03) and it was confirmed by the model adjusted for the duration of hospitalisation (p = 0.04). Yet day-care ward patients had a higher number of rehospitalisations within one year after discharge. CONCLUSIONS: The absolute superiority of treatment in an inpatient ward over day-care setting in terms of its effectiveness was not confirmed using BPRS, MANSA scores and index of rehospitalisation. PMID- 19189602 TI - [Association of the tyrosine hydroxylase gene polymorphism with schizophrenia in the population of central Poland]. AB - AIM: The aim of the study was a comparative analysis of the polymorphism in TH01 locus-- tetranucleotide microsatellite region located in the first intron of the tyrosine hydroxylase gene (TH) between a group of Polish patients suffering for schizophrenia and their regionally matched healthy subjects. METHOD: One hundred patients affected by paranoid schizophrenia and healthy individuals with negative family history of psychiatric disorders as control, were investigated. Genomic DNA was isolated from peripheral blood, amplification of TH01 locus was carried out using the following sequences of primers: 5'-GTG GGC TGAAAAGCT CCCGAT TAT-3', 5'-ATT CAA AGG GTA TCT GGG CTC TGG-3', PCR products were detected on the ABI Prism 377 sequencer. Distributions of alleles, genotypes and homo-heterozygosity of the patients were compared with those of the matching controls using the RXC program created by G. Carmody. Relative Risk (RR) of the disease was calculated according to the formula given by Dyer & Werrens. RESULTS: . The conducted analysis showed the existence of statistically significant differences in the distribution of alleles, as well as genotypes between the schizophrenics and the control population. We revealed that 7 allele is present statistically significantly more often in the group of patients and its presence increases the risk of schizophrenia almost twofold (RR = 1.89). Whereas the presence of 9.3 allele reduces the risk of the disease (RR = 0.72), in the homozygote form 9.3 9.3 even over three times (RR = 0 .32). CONCLUSION: The revealed differences in the susceptibility to schizophrenia depending on polymorphic allele variants in repetitive TCAT sequence in TH01 locus may be associated with the function of a regulatory element in the process of TH gene transcription. Our findings need further investigation in a larger sample of patients. PMID- 19189603 TI - [The assessment of the mental state of patients during simultaneous treatment with psychotropic drugs, antipsychotics included, and bromocriptine]. AB - AIM: The aim of this study was to assess the mental state of patients treated with psychotropic drugs (LPT) and bromocriptine (BRC) at the same time. METHODS: 25 female patients in the stable mental state treated with antipsychotics (LPP) (13 patients were also treated with other LPT) with an average age of 25.56 years were included in the study. All patients presented clinical symptoms of HPRL: menstrual disturbances or galactorrhea and their serum PRL was increased or the test with metoclopramide was incorrect. BRC (1.25-8.75 mg per day) was ordered to eliminate HPRL and observation was conducted over a period of 3 months. Two weeks, a month, 2 months and 3 months after the BRC treatment had been started, an evaluation of the mental state of the patients was conducted. The patients' mental state was evaluated with the Clinical Global Impression Scale (CGI), the Hamilton Depression Rating Scale (HDRS) and the Positive and Negative Syndrome Scale (PANSS). RESULTS: After 3 months of the study in the whole group there was no significant change in the mental state evaluated with the CGI and the HDRS. There was a worsening in PANSS (p < 0.05) which was mainly due to the worsening (p < 0.01) in the subscale of general psychiatric symptomatology of PANSS (PANSS G). There was no significant change in the subscales of positive and negative symptoms of PANSS (PANSS-P and PANSS-N respectively). CONCLUSIONS: The results show that during the 3 months of the BRC treatment there was no worsening in positive, negative and affective symptoms. The study suggests that adding BRC may be a safe option of management of psychotropic-induced HPRL in some psychiatrically stable patients, although further studies are necessary. PMID- 19189604 TI - [Disease--defence--manipulation: the difficulties in providing forensic psychiatry opinions]. AB - The challenges met by the psychiatrist and the psychologist and the difficulties in providing forensic-psychiatry and forensic-psychology opinions have been reviewed, based on examples. The studied patient was hospitalised 10 times and the forensic-psychiatry opinion passed 15 times during the judiciary process. Different psychiatric diagnoses were made and different soundness of mind were passed. The psychiatric health status were expresses by professors of psychiatry who did not participate directly in passing the forensic-psychiatry opinion. The studied patient was examined by other specialists and assessed by certificating medical doctors with the aim of getting disability pension benefits. The effect of medical certificates and testimonials from different medical doctors were analysed. Analysing this example, revealed the problem of proper formulation of content of medical documents to support the medical diagnosis and declared soundness of mind during the passing of forensic-psychiatry opinion. The doctors treating the patients and doctors passing opinion on the treated patient had a different assessment of the diagnoses and soundness of mind for the studied patients. Irrespective of the immediate aim of the examinations, all professionals providing assessment should mind the consequences of opinions passed by them and especially, the possibility of the opinion being used by the subject to prolong the judiciary process or even avoid legal responsibility. The independence (sovereignty) of the expert requires consideration in the context of prior multiple forensic-psychiatry opinion leaders in the field of psychiatry and the need for the expert to assume an attitude towards these opinions. PMID- 19189605 TI - Global, regional, or...? PMID- 19189606 TI - Fostering disaster resilient communities across the globe through the incorporation of safe and resilient hospitals for community-integrated disaster responses. AB - The impact of catastrophic events on hospitals and communities is huge and continues to hinder progress in developing nations and industrialized countries alike. Over the last 10 years, the UN/ISDR has sponsored a series of global conferences to increase awareness of the importance of risk and vulnerability reduction and the need to build disaster resilient communities. In recognition that hospitals contribute to the health and resiliency of a community, ISDR has adopted the PAHO and WHO "Safe and Resilient Hospital" initiative. The primary focus of the initiative is to ensure the physical and functional integrity of hospitals during a disaster. Hospital resiliency also must encompass the ability to fully integrate hospital facilities and their services into an overall community response to prevent hospitals from becoming isolated from other responding organizations. In order to help promote the "safe and resilient hospital" initiative, during the 15WCDEM, three strategic objectives were identified for hospitals that meet SEARO Benchmark #5. These are: (1) establish tiers of standards (criteria) that define "safe and resilient" hospitals in diverse regions of the world; (2) develop a tool to assess the extent to which hospitals, meet the criteria for "safe and resilient" hospitals; and (3) apply the evidence derived from use of this tool to promote the concept of "safe and resilient" hospitals as an integral part of emergency preparedness, responses, and recovery, and maximize the political commitment from decision-makers within and outside the healthcare sector to support, protect, and integrate the initiative into a community-wide disaster response. Ultimately, attaining these objectives will protect the lives of patients and healthcare workers, ensure that hospitals are able to provide urgently needed and everyday medical care to the community they serve and minimize risk and vulnerabilities of patients, healthcare workers and other individuals within the community. PMID- 19189607 TI - Revisiting blood transfusion preparedness: experience from the Bam earthquake response. AB - Blood transfusion plays a critical role in the provision of medical care for disasters due to man-made and natural hazards. Although the short-term increase in blood donations following national disasters is well-documented, some aspects of blood transfusion during disasters remain under study. The 2003 earthquake in Bam, Iran resulted in the death of >29,000 people and injured 23,000. In total, 108,985 blood units were donated, but only 21,347 units (23%) actually were distributed to hospitals around the country. Kerman Province, the site of the disaster, received 1,231 (1.3%) of the donated units in the first four days after the disaster. The Bam experience revealed crucial missteps in the development of a post-event strategy for blood product management, and led to the development of a detailed disaster preparedness and response plan that addresses issues of donation, distribution, communication, transportation, and coordination. The current plan requires the Iranian Blood Transfusion Organization to convene a disaster task force immediately as the main coordinator of all disaster preparedness and response activities. PMID- 19189608 TI - Blood: the ultimate donation in emergencies. PMID- 19189609 TI - Cognitive-behavioral psychology: implications for disaster and terrorism response. AB - Given the personal and societal costs associated with acute impairment and enduring post-traumatic stress disorder (PTSD), the mental health response to disasters is an integral component of disaster response planning. The purpose of this paper is to explore the compatibility between cognitive-behavioral psychology and the disaster mental health model, and explicate how cognitive behavioral perspectives and intervention methods can enhance the effectiveness of disaster mental health services. It is argued that cognitive-behavioral methods, if matched to the contexts of the disaster and the needs of individuals, will improve efforts to prevent the development of PTSD and other trauma-related problems in survivors of disaster or terrorist events. First, the similarities between models of care underlying both disaster mental health services and cognitive-behavioral therapies are described. Second, examples of prior cognitive behavioral therapy-informed work with persons exposed to disaster and terrorism are provided, potential cognitive-behavioral therapy applications to disaster and terrorism are explored, and implications of cognitive-behavioral therapy for common challenges in disaster mental health is discussed. Finally, steps that can be taken to integrate cognitive-behavioral therapy into disaster mental health are outlined. The aim is to prompt disaster mental health agencies and workers to consider using cognitive-behavioral therapy to improve services and training, and to motivate cognitive-behavioral researchers and practitioners to develop and support disaster mental health response. PMID- 19189610 TI - Profile of emergency medical dispatch calls for breathing problems within the medical priority dispatch system protocol. AB - INTRODUCTION: A common chief complaint to emergency dispatch communication centers worldwide is "breathing problems". The chief complaint of breathing problems represents a wide spectrum of underlying diseases, patient conditions, and onset types. The current debate is on the potential ability of a dispatch protocol to safely and with high specificity, differentiate patients with minor or non-critical conditions from those conditions that pose risk to the patient and require advanced life support evaluation and care. This issue also has extended into the paramedic prehospital evaluation realm. OBJECTIVE: The objective of this study was to describe the distribution of Medical Priority Dispatch System (MPDS) codes representing the spectrum of clinical descriptions within the breathing problems chief complaint and their associated outcomes, at the scene and during transport, as determined by [UK] paramedics. METHODS: A retrospective, one-year study (September 2005 to August 2006) of a de-identified aggregate dataset from the London Ambulance Service (LAS) Trust was evaluated. A profile of the distribution of calls, incidents, patients, and outcomes (cardiac arrest [CA] and blue-in [BI] high acuity i.e., patients transported with lights and siren based on paramedic protocol) for the breathing problems chief complaint was evaluated. Odds ratios and 95% confidence intervals (CI) were used to quantify associations between the MPDS priority level's concurrent asthmatic conditions and outcomes. Two-sided Fisher's exact p-values were obtained to determine statistically significant associations, at a level of0.05. RESULTS: Sixteen percent (95,848/599,093) of all the patients were classified under the breathing problems chief complaint. Of these 95,848 patients, 367 (0.38%) were CA outcomes, and 7.82% (n = 7,493) were BI outcomes.The Cardiac Arrest Quotient (i.e., the number of CA cases as a percentage of the number of patients) for the ECHO priority level was 46 times higher than was that of non-ECHO priority levels: DELTA and CHARLIE (17.05% vs. 0.37%). Asthmatics were associated with CA outcome (OR(95%CI): 0.60(0.47,0.77), p <0.001), but not with BI outcome. CONCLUSIONS: The MPDS coding yielded a richer mix of severe outcomes in the higher priority levels.The Severe Respiratory Distress coding had the greatest number of patients and severe outcomes. Future studies that help refine the Severe Respiratory Distress code in the MPDS by more specific subgroups of patients would be beneficial. PMID- 19189611 TI - Development of prehospital, population-based triage-management protocols for pandemics. AB - The lack of disease-specific triage-management protocols that address the unique aspects of a pandemic places emergency medical services, and specifically, emergency medical services practitioners, at great risk. Without adequate protocols, the emergency health system will risk needless exposure, loss of functional capacity, and inappropriately triaged patients. This paper reports on the development of population-based triage-management protocols at two patient points of contact. The primary objective of the triage-management protocols is to identify patients infected by or exposed to the biological agent, and consequently, appropriately triage patients so as to optimize the utilization of emergency medical services and surge capacity resources through disposition and care at hospital- and non-hospital-based care facilities. Protocols must include standardized "flu questions" and a Fear and Resiliency Checklist to ensure protection and separation of the susceptible population from those infected or exposed. PMID- 19189612 TI - Characteristics of emergency medical technicians involved in ambulance crashes. AB - OBJECTIVES: This study utilizes a [US] national sample of emergency medical services (EMS) professionals to explore the hypothesis that demographic and work related characteristics are associated with involvement in ambulance crashes. METHODS: In 2004, a cohort of nationally registered EMS professionals was surveyed to determine ambulance crash involvement during a 12-month period. Involvement in an ambulance crash was the outcome variable of interest. Demographics such as age, community size, service type, call volume, time spent in an ambulance, and current sleep problems were analyzed as independent variables. A multivariate logistic regression model identified variables associated with involvement in an ambulance crash within the past year. RESULTS: Surveys were received from 1,775/5,565 (32.0%) participants; 1,297 (73.1%) met the inclusion criteria. A total of 111 (8.6%) of participants reported being involved in an ambulance crash within the past 12 months. When controlling for call volume and time in an ambulance, the odds of involvement in an ambulance crash within the past year were significantly higher for younger EMS professionals and those reporting sleep problems. CONCLUSIONS: Results from this analysis suggest age and sleep problems are associated with involvement in an ambulance crash. Future studies should investigate interventions to minimize the effects of these associations. PMID- 19189613 TI - Evaluation of online disaster and emergency preparedness resources. AB - INTRODUCTION: Increasingly, individuals are relying on the Internet as a major source of health information. When faced with sudden or pending disasters, people resort to the Internet in search of clear, current, and accurate instructions on how to prepare for and respond to such emergencies. Research about online health resources ascertained that information was written at the secondary education and college levels and extremely difficult for individuals with limited literacy to comprehend. This content analysis is the first to assess the reading difficulty level and format suitability of a large number of disaster and emergency preparedness Web pages intended for the general public. OBJECTIVES: The aims of this study were to: (1) assess the readability and suitability of disaster and emergency preparedness information on the Web; and (2) determine whether the reading difficulty level and suitability of online resources differ by the type of disaster or emergency and/or Website domain. METHODS: Fifty Websites containing information on disaster and/or emergency preparedness were retrieved using the Google search engine. Readability testing was conducted on the first Web page, suggested by Google, addressing preparedness for the general public. The reading level was assessed using Flesch-Kincaid (F-K) and Flesch Reading Ease (FRE) measures. The Suitability Assessment of Materials (SAM) instrument was used to evaluate additional factors such as graphics, layout, and cultural appropriateness. RESULTS: The mean F-K readability score of the 50 Websites was Grade 10.74 (95% CI = 9.93, 11.55). The mean FRE score was 45.74 (95% CI = 41.38, 50.10), a score considered "difficult."A Web page with content about both risk and preparedness supplies was the most difficult to read according to F-K (Grade level = 12.1). Web pages with general disaster and emergency information and preparedness supplies were considered most difficult according to the FRE (38.58, 95% CI = 30.09, 47.08). The average SAM score was 48% or 0.48 (95% CI = 0.45, 0.51), implying below average suitability of these Websites. Websites on pandemics and bioterrorism were the most difficult to read (F-K: p = 0.012; FRE: p = 0.014) and least suitable (SAM: p = 0.035) compared with other disasters and emergencies. CONCLUSIONS: The results suggest the need for readily accessible preparedness resources on the Web that are easy-to-read and visually appropriate. Interdisciplinary collaborations between public health educators, risk communication specialists, and Web page creators and writers are recommended to ensure the development and dissemination of disaster and emergency resources that consider literacy abilities of the general public. PMID- 19189614 TI - Evidence-based approach for disaster preparedness authorities to inform the contents of repositories for prescription medications for chronic disease management and control. AB - INTRODUCTION: Chronic diseases are major causes of death and disability and often require multiple prescribed medications for treatment and control. Public health emergencies (e.g., disasters due to natural hazards) that disrupt the availability or supply of these medications may exacerbate chronic disease or even cause death. PROBLEM: A repository of chronic disease pharmaceuticals and medical supplies organized for rapid response in the event of a public health emergency is desirable. However, there is no science base for determining the contents of such a repository. This study provides the first step in an evidence based approach to inform the planning, periodic review, and revision of repositories of chronic disease medications. METHODS: Data from the 2004 National Hospital Ambulatory Medical Care Survey (NHAMCS) were used to examine the prescription medication needs of persons presenting to US hospital emergency departments for chronic disease exacerbations. It was assumed that the typical distribution of cases for an emergency department will reflect the patient population treated in the days after a public health emergency. The estimated numbers of prescribed drugs for chronic conditions that represent the five leading causes of death, the five leading primary diagnoses for physician office visits, and the five leading causes of disease burden assessed by disability adjusted life years are presented. RESULTS: The 2004 NHAMCS collected data on 36,589 patient visits that were provided by 376 emergency departments. Overall, the five drug classes mentioned most frequently for emergency department visits during 2004 were narcotic analgesics (30.7 million), non-steroidal anti inflammatory drugs (25.2 million), non-narcotic analgesics (15.2 million), sedatives and hypnotics (10.4 million), and cephalosporins (8.2 million). The drug classes mentioned most frequently for chronic conditions were: (1) for heart disease, antianginal agents/vasodilators (715,000); (2) for cancer, narcotic analgesics (53,000); (3) for stroke, non-narcotic analgesics (138,000); (4) for chronic obstructive pulmonary disease, anti-asthmatics/bronchodilators (3.2 million); and (5) for diabetes, hypoglycemic agents (261,000). Ten medication categories were common across four or more chronic conditions. CONCLUSIONS: Persons with chronic diseases have an urgent need for ongoing care and medical support after public health emergencies. These findings provide one evidence based approach for informing public health preparedness in terms of planning for and review of the prescription medication needs of clinically vulnerable populations with prevalent chronic disease. PMID- 19189615 TI - Pharmaceuticals and chronic diseases in disaster preparedness. PMID- 19189616 TI - Acute stress symptoms, dissociation, and depression among rescue personnel 24 hours after the Bet-Yehoshua train crash: the effects of exposure to dead bodies. AB - INTRODUCTION: The immediate impact of exposure to severe wounds, dead bodies, and immediate threat to life has been understudied. Most studies focus on the acute stress disorder and/or post-traumatic stress disorder phases in order to assess rescue personnel's symptomatology, and tend to neglect the immediate exposure to elements of the disaster. HYPOTHESIS: Rescue personnel who had a history of previous exposure to dead bodies would exhibit higher levels of acute stress symptoms, dissociation, and depressive symptoms within the 24 hours following a traumatic event. METHODS: Twenty-three rescue personnel participated in the search and excavation of dead and mutilated bodies following the Bet-Yehoshua train crash in Israel. The rescue personnel group was divided based on previous exposure to dead bodies. Each participant completed a demographic questionnaire, which included a question on perceived threat to life, the impact of event scale revised, the dissociative experience scale, and the center of epidemiologic studies depression questionnaire. Student's t-tests, along with multivariate analysis of covariance (MANCOVA) were conducted in order to learn which factors are related to psychiatric symptomatology following the immediate exposure to such stressors. RESULTS: Among rescue personnel, those with previous exposure to dead bodies did not differ in their levels of acute stress symptoms, dissociation, and depressive symptoms from those who were not previously exposed to dead bodies. CONCLUSIONS: These results may suggest the possibility that the impact of exposure to dead bodies does not emerge in the acute stress reactions (ASR) phase (up to 24 hours after the event), but later when people have time to process the trauma. Another possibility is that the rescue coping mechanisms of detachment may serve as a buffer for the horrific sights encountered during the ASR period. PMID- 19189617 TI - Model for preparedness of a public institution for a terrorist attack. AB - Public institutions such as governmental facilities, hospitals, universities, and amusement parks may be targeted by terrorists using weapons of mass destruction due to their potential to cause large numbers of casualties. Consequentially, these institutions should be prepared to manage such an event by the development and implementation of specific preparedness guidelines for any conventional or unconventional terrorist attacks. In order to test the preparedness of such an institution for a chemical event, a large-scale drill focusing on the first medical team to respond was conducted. Some important lessons regarding the way the medical team operates and communicates were learned from this drill. Periodic drills should be performed in order to assess the practicality and applicability of these guidelines. PMID- 19189618 TI - KAMEDO Report No. 91: 2004 tsunami disaster in Asia--home transport and emergency care in Sweden. AB - This is a descriptive report of the Swedish authorities' responses to the tsunami that affected Southeast Asia in December 2004. The main focus is the care of survivors and the injured during their transportation from Thailand and their return to Sweden. The psychological and physical after-effects also are presented based on a poll conducted one year after the tsunami. PMID- 19189619 TI - KAMEDO Report No. 92: Evacuation of Swedes from Lebanon--observational studies in connection with the war in Lebanon in summer 2006. AB - A large-scale armed conflict between Hezbollah and Israel in July 2006 caused the evacuation of a large number of Swedish residents from Lebanon. This report describes the evacuation throughout its various stages. Swedish authorities were prepared for an event of this type from the experience of the 2004 Asian tsunami disaster. Lessons learned from the management and medical care during the evacuations are presented in this report. PMID- 19189620 TI - Infection control. Zeroing in on infection prevention and control. PMID- 19189621 TI - Construction. Building for the future. PMID- 19189622 TI - Supply chain. Cost of goods grab executives' attention. PMID- 19189623 TI - Designs for the delicate: a look at evolving NICU design standard. PMID- 19189624 TI - Office work: the ES professional's role in MOB cleaning and disinfection. PMID- 19189625 TI - [A magical year: further training in pediatric grief support]. PMID- 19189626 TI - [The sibling bed: recent aspects of nursing research]. PMID- 19189627 TI - [Primary nursing of extremely low birth weight premature infants: with special reference to skin care]. PMID- 19189628 TI - [Familiarity with craniosacral nursing in pediatric nursing]. PMID- 19189629 TI - [Vaccination against nosocomial infections: wishful thinking or possible clinical reality?]. PMID- 19189630 TI - ["Selective mutism--when children are speechless"]. PMID- 19189631 TI - [Vitamin C and zinc tablets do not prevent or cure the common cold: scientific proof of the efficacy of food supplements is absent]. PMID- 19189632 TI - [In the hospital test: germicidal copper. Metal coatings on door knobs and light switches to prevent infections]. PMID- 19189633 TI - [Born to learn: optimizing the brain by early education]. PMID- 19189634 TI - [Disease notification in diarrhea]. PMID- 19189635 TI - The impact of enzymatic degradation on the uptake of differently sized therapeutic molecules. AB - BACKGROUND: The extracellular matrix represents a major barrier for drug delivery. This work compares the effects of collagenase and hyaluronidase on tumour uptake and distribution of two differently sized therapeutic molecules, IgG and liposomal doxorubicin. MATERIALS AND METHODS: The enzymes were injected i.v. prior to the therapeutic molecules, and uptake and distribution were studied by confocal laser scanning microscopy. The therapeutic molecules were colocalized with the vasculature and collagen network visualized by the second harmonic signal. RESULTS: Hyaluronidase increased the uptake of liposomal doxorubicin to a small extent, whereas collagenase had no effect. Collagenase increased, but hyaluronidase reduced the uptake of IgG. Neither of the enzymes induced changes in the collagen network measured by the second harmonic signal. CONCLUSION: Degradation of the collagen network improves delivery of molecules with the size of IgG, whereas degradation of the gel of glycosaminoglycans has a higher impact on the distribution of small drugs such as doxorubicin. PMID- 19189636 TI - Alpha-folate receptor expression in epithelial ovarian carcinoma and non neoplastic ovarian tissue. AB - BACKGROUND: The goal of this study was to evaluate the expression pattern and intracellular localization of alpha-folate receptor (alpha-FR) protein in human ovarian carcinoma compared with non-neoplastic ovarian tissue. MATERIALS AND METHODS: Using immunohistochemistry (IHC), alpha-FR protein expression was analyzed in specimens of 104 human ovarian carcinomas and 30 non-neoplastic ovaries. RESULTS: In 97% of the ovarian carcinomas, clear alpha-FR protein expression was detected (14% weak, 39% moderate, 44% strong). In the non neoplastic ovaries, no (37%) or only weak (63%) expression was observed (p<0.0001). The tumor cells were characterized by a diffuse and homogeneous staining pattern. In tumor and non-tumor tissue, alpha-FR protein was detected predominantly in the cellular cytoplasm. In 41% of the ovarian carcinomas, cytoplasmic expression was localized towards the outer boarders of the invasive tumor cells and 30% exhibited additional nuclear alpha-FR protein expression. CONCLUSION: Compared with nonneoplastic ovaries, alpha-FR protein is overexpressed in human ovarian carcinoma tissue. PMID- 19189637 TI - Identification of parental chromosomes involved in translocations BCR-ABL, t(9;22) and PML-RARA, t(15;17). AB - Cells of blood and bone marrow often exhibit a genome- or ploidywise organization of the two haploid sets, representing apparently maternal and paternal chromosomes in interphase nuclei and in metaphase spreads. This provides the opportunity to perform "genomic karyotyping." Such application of karyotyping may indicate whether two chromosomes involved in a translocation are both maternal, both paternal, or intermingled, i.e., one maternal and the other paternal (we refer to this as mixed). The parental origin for these translocations likely has profound differences and implications in disease expression and response to treatments, making such information very important to personalized medicine. In this mini-review, we present our observations from specimens with translocations BCR-ABL, t(9;22) and PML-RARA, t(15;17). About 20% metaphases of these specimens indicated ploidywise organization and were amenable to genomic karyotyping analysis. Fluorescence in situ hybridization (FISH) probes for BCR-ABL translocation suggest a close approximation of the HSA 9 and 22, as control values for false-positive signals run from approximately 5-10%. Given a ploidywise distribution of the maternal and paternal sets of chromosomes, it would be expected that the chromosomes involved in the translocation t(9;22) would more often belong to one of the two genomes, either maternal or paternal. Contrastingly, HSA 15 and 17 are not considered as spatially close to each other and therefore an intragenomic involvement would be rarer for translocation t(15;17). In 14 out of the 21 (66.6%) specimens with informative metaphases, the chromosomes involved in the translocation BCR-ABL were restricted to one of the two genomes--either maternal or paternal. In cases of translocation PML-RARA only 4 out of 21 (19.1%) specimens indicated an intragenomic involvement. These simple yet informative analyses of cancer-related translocations show profound underlying genomic origins and lend support to genomic karyotyping. PMID- 19189639 TI - Pioglitazone, a ligand for peroxisome proliferator-activated receptor-gamma acts as an inhibitor of colon cancer liver metastasis. AB - BACKGROUND: Peroxisome proliferator-activated receptor-gamma (PPARgamma) is a member of the steroid receptor superfamily. Liganded PPARgamma can inhibit cancer cell proliferation. The in vitro and in vivo inhibitory effect of the synthetic ligands, ciglitazone (CGZ) and pioglitazone (PGZ), on human colon cancer was investigated. MATERIALS AND METHODS: Cell proliferation and the expression of PPARgamma, cyclooxygenase (COX)-2 and cyclin D1 were assessed in colon cancer cells treated with CGZ or PGZ. After subcutaneous or splenic inoculation of severe combined immunodeficient (SCID) mice using colon cancer HT-29 and SW480 cells, PGZ was administered orally and tumor growth inhibition was assessed by xenograft volume. The COX-2, cyclin D1 and PPARgamma expression in the HT-29 cells was evaluated. RESULTS: Cultured HT-29 and SW480 cells expressed PPARgamma and proliferation was inhibited by CGZ and PGZ. Oral PGZ inhibited xenograft tumor growth and liver metastases in the SCID mouse and suppressed expression of COX-2 and cyclin D1 in HT-29 cells. CONCLUSION: PGZ down-regulates COX-2 and cyclin D1 and inhibits colon cancer proliferation and liver metastasis, making PPARgamma a candidate target for the treatment/prevention of colon cancer metastasis. PMID- 19189640 TI - Radioprotection of murine gastrointestinal epithelium by interleukin-1alpha involves down-regulation of the apoptotic response. AB - BACKGROUND: Interleukin-1alpha (IL-1) is known to radioprotect the gastrointestinal tract, but the mechanism by which this protection occurs remains unclear. These studies were undertaken to investigate whether the radioprotective potential of IL-1 may be linked to an ability to reduce apoptosis within the gastrointestinal crypts. MATERIALS AND METHODS: IL-1 was administered to C57Bl/6 mice 24 hours prior to receiving 8 Gy abdominal X-irradiation (xRT). At designated times, experimental mice were sacrificed, jejunal tissue removed, and paraffin-embedded sections analyzed for apoptosis indices (AI) and immunohistochemical determination of active caspase-3, -8 and -9. RESULTS: AI data demonstrated that 8 Gy irradiation resulted in a marked jejunal apoptotic response, but IL-1 pretreatment significantly attenuated this response. Concomitant with this attenuation, reduced levels of caspase-3 and 9, but not caspase-8, activation were observed, particularly within goblet cells. CONCLUSION: The results outlined herein suggest that radioprotection by IL-1 is mediated, at least in part, through a reduction in the apoptotic response which appears to involve down-regulation of the intrinsic apoptotic pathway. PMID- 19189638 TI - Role of caspases in 5-FU and selenium-induced growth inhibition of colorectal cancer cells. AB - BACKGROUND: The mechanisms that could explain the poor sensitivity to 5-FU in certain colorectal cancer (CRC) cells were investigated and whether or not cotreatment with low doses of selenium would offer a therapeutic benefit was explored. MATERIALS AND METHODS: Four CRC cell lines (Caco2, RKO, DLD1 and HT-29) with defined tumor signatures and seven different chemical forms of selenium were tested. RESULTS: 5-FU partially inhibited the HT-29 and RKO cells, but had a weak effect on the DLD1 and almost none on the Caco2 cells. Selenous acid and sodium selenite induced growth inhibition of the DLD1, RKO and HT-29 cells, with a marginal effect on the Caco2 cells. The Caco2 cells with mutant p53, failure to activate caspase-8, -9, -7 and -3 and with hypermethylated caspase-8 were resistant to 5-FU. Conversely, RKO cells expressing wildtype p53, proteolytically activated caspase-8, -9, -7 and -3 and unmethylated caspase-8 were more responsive to 5-FU and selenous acid-induced apoptosis. CONCLUSION: Combination treatment with selenous acid may offer an efficacious strategy to overcome 5-FU resistance in certain CRC cells. PMID- 19189641 TI - Genomic profile of colon cancer metastases. AB - BACKGROUND: In colon cancer, the occurrence of metastases is associated with microsatellite stability. As metastatic cells derive from a clonal expansion of primary tumor cells, specific genomic alterations are expected in addition to the common genomic profile. PATIENTS AND METHODS: Genome-wide allelotyping was performed on 75 liver metastases samples from sporadic colon cancer. RESULTS: No microsatellite instability was observed. Allelic loss on 5q in metastases was significantly different from that of non metastatic primary tumors (16/58 vs. 43/75, p=0.0008). Four additional chromosomes, 4, 7, 8 and 19, were more frequently lost in liver metastases, but statistical significance was reached only for 19q (14/63 versus 2/68 in primary tumors; p=0.033 after Dunn-Sidak adjustment). CONCLUSION: This study confirms that liver metastasis is rather restricted to patients with microsatellite stable colon cancer and these retain the 5q arm with high frequency. In addition, it suggests that loss of 19q may be critical for one of the steps involved in the development of liver metastases. PMID- 19189642 TI - PTEN mutation: many birds with one stone in tumorigenesis. AB - The PTEN (phosphatase and tensin homolog deleted on chromosome ten) tumor suppressor gene is mutated in a wide range of malignancies and recent studies have demonstrated that PTEN prevents tumorigenesis through multiple mechanisms. PTEN functions as a plasma-membrane lipid phosphatase that antagonizes the PI3K (phosphoinositide 3 kinase)-AKT pathway. PTEN physically and genetically interacts with the central genome guardian p53. PTEN also associates with the centromeric protein CENP-C to maintain centromere integrity and suppresses chromosomal instability from DNA double-strand breaks (DSBs) through transcriptional regulation of Rad51 (radiosensitive yeast mutant 51). Moreover PTEN controls the growth and proliferation of haematopoietic stem cells (HSC) and restrains cells from leukemia in an mTOR (mammalian target of rapamycin) dependent manner. Thus, restoring PTEN functions in cancer cells directly or indirectly holds great promise for cancer therapy. PMID- 19189643 TI - Exploitation of the Notch signaling pathway as a novel target for cancer therapy. AB - The Notch signaling pathway appears to be responsible for maintaining a balance between cell proliferation and apoptosis and thus it has been suggested that Notch may play an important role in species development and in the development and progression of several malignancies. Therefore, the Notch signaling pathway may represent a novel therapeutic target, which could have the highest therapeutic impact in modern medicine. This review describes the mechanisms of signal transduction of the Notch signaling pathway and provides emerging evidence in support of its role in the development of human malignancies. Further attempts have been made to summarize the role of several chemopreventive agents that could be useful for targeted inactivation of Notch signaling, which could become a novel approach for cancer prevention and treatment. PMID- 19189644 TI - Dietary sphingomyelin inhibits colonic tumorigenesis with an up-regulation of alkaline sphingomyelinase expression in ICR mice. AB - BACKGROUND: Sphingomyelin (SM) hydrolysis generates biologically active products regulating cell growth, differentiation and apoptosis. Dietary SM has been found to inhibit colonic tumorigenesis. Alkaline sphingomyelinase (alk-SMase) is the key enzyme responsible for sphingomyelin digestion in the gut. Whether or not dietary sphingomyelin affects alk-SMase expression was examined in a colon cancer animal model. MATERIALS AND METHODS: Imprinting control region (ICR) mice were injected with 1,2-dimethylhydrazine (DMH) and then fed a diet with or without SM (0.5 g/kg in diet) for 22 weeks. The colonic tumorigenesis and alk-SMase activity were determined and alk-SMase expression was examined by Western blot and PCR. RESULTS: Dietary SM inhibited the tumorigenesis and increased the alk-SMase activity in the colon by 65%. The increased activity was associated with increased enzyme protein and mRNA expression. No changes of acid and neutral sphingomyelinase activities were found. CONCLUSION: Long-term supplementation with dietary sphingomyelin up-regulates colonic alk-SMase expression, which may contribute to the inhibitory effects of sphingomyelin against colonic carcinogenesis. PMID- 19189645 TI - Combined oncolytic virotherapy with herpes simplex virus for oral squamous cell carcinoma. AB - BACKGROUND: The effect of dual infection with herpes simplex virus type 1 (HSV-1) mutants on human oral squamous cell carcinoma (SCC) cells was examined. MATERIALS AND METHODS: Human oral SCC cells were infected with gamma1(34.5) gene-deficient HSV-1 R849 and HSV-1 HF that has multiple mutations and induces cell fusion. Cell viability was measured by LDH release assay. Athymic mice were injected with oral SCC cells into the buccal region to induce subcutaneous tumors. RESULTS: Oral SCC cells were infected with R849, followed by infection with R849 or HF. Virus production was elevated by both strains of HSV-1. Although the release of LDH from R849-infected cells was increased by secondary infection with R849 or HF, the effect of HF was more remarkable. When nude mouse tumors were treated with R849, HF, R849+R849, or R849+HF, treatment with R849+HF was the most effective. CONCLUSION: These results suggest that fusion-inducing virus HF enhances the oncolytic ability of gamma1(34.5) gene-deficient HSV-1 and provides a rationale for using fusogenic viruses as enhancing agents PMID- 19189646 TI - The guggul for chronic diseases: ancient medicine, modern targets. AB - Identification of active principles and their molecular targets from traditional medicine is an enormous opportunity for modern drug development. Gum resin from Commiphora wightii (syn C. mukul) has been used for centuries in Ayurveda to treat internal tumors, obesity, liver disorders, malignant sores and ulcers, urinary complaints, intestinal worms, leucoderma (vitiligo), sinuses, edema and sudden paralytic seizures. Guggulsterone has been identified as one of the major active components of this gum resin. This steroid has been shown to bind to the farnesoid X receptor and modulate expression of proteins with antiapoptotic (IAP1, XIAP, Bfl-1/A1, Bcl-2, cFLIP, survivin), cell survival, cell proliferation (cyclin D1, c-Myc), angiogenic, and metastatic (MMP-9, COX-2, VEGF) activities in tumor cells. Guggulsterone mediates gene expression through regulation of various transcription factors, including NF-kappaB, STAT-3 and C/EBPalpha, and various steroid receptors such as androgen receptor and glucocorticoid receptors. Modulation of gene expression by guggulsterone leads to inhibition of cell proliferation, induction of apoptosis, suppression of invasion and abrogation of angiogenesis. Evidence has been presented to suggest that guggulsterone can suppress tumor initiation, promotion and metastasis. This review describes the identification of molecular targets of guggulsterone, cellular responses to guggulsterone, and animal studies and clinical trials of guggulsterone in cancer and other diseases. PMID- 19189647 TI - New method for separation of subpopulations from a heterogeneous colon cancer cell line. AB - BACKGROUND: To understand the heterogeneity of human colon cancers, a new method to separate cancer subpopulations was developed. MATERIALS AND METHODS: Cells from a human colon cancer cell line, DLD-1, were seeded on an 8 microm pore membrane. After six hours, the cells which remained beneath the membrane as well as the cells which dropped onto the 24-well plate were collected. To clarify the differences between the two subpopulations, transepithelial electrical resistance (TEER) and immunocytochemistry were evaluated. RESULTS: Two subpopulations, clones D and A, were separated from DLD-1 with the newly developed method. Both subpopulations showed quite different TEER values and different arrangements of cell-cell contact. In addition, the distinct subcellular localizations of claudin family proteins and zonula occludens-1 (ZO-1) were identified. CONCLUSION: A new separation method to isolate colon cancer subpopulations was established in which the intercellular junctions differed. This method can be considered as a helpful tool in the investigation of colon cancer heterogeneity. PMID- 19189648 TI - Green tea catechin inhibits fatty acid synthase without stimulating carnitine palmitoyltransferase-1 or inducing weight loss in experimental animals. AB - BACKGROUND: The enzyme fatty acid synthase (FASN) is highly expressed in many human carcinomas and its inhibition is cytotoxic to human cancer cells. The use of FASN inhibitors has been limited until now by anorexia and weight loss, which is associated with the stimulation of fatty acid oxidation. MATERIALS AND METHODS: The in vitro effect of (-)-epigallocatechin-3-gallate (EGCG) on fatty acid metabolism enzymes, on apoptosis and on cell signalling was evaluated. In vivo, the effect of EGCG on animal body weight was addressed. RESULTS: EGCG inhibited FASN activity, induced apoptosis and caused a marked decrease of human epidermal growth factor receptor 2 (HER2), phosphatidylinositol 3-kinase (PI3K)/AKT and extracellular (signal)-regulated kinase (ERK) 1/2 proteins, in breast cancer cells. EGCG did not induce a stimulatory effect on CPT-1 activity in vitro (84% of control), or on animal body weight in vivo (99% of control). CONCLUSION: EGCG is a FASN inhibitor with anticancer activity which does not exhibit cross-activation of fatty acid oxidation and does not induce weight loss, suggesting its potential use as an anticancer drug. PMID- 19189650 TI - Radioiodinated methylene blue--a promising agent for melanoma scintigraphy: labelling, stability and in vitro uptake by melanoma cells. AB - Melanoma is a tumor of continuously increasing incidence for which new methods of imaging and targeted therapy are widely sought. Radioiodinated methylene blue is a promising tracer, showing selective uptake in human pigmented melanoma cells. We performed 131I-labeling of the tracer using 1% methylene blue injection United States Pharmacopeia (USP) and 131I sodium iodide. For quality control, a Merck high performance liquid chromatography (HPLC) system was used. We developed a new HPLC procedure using 0.1% trifluoroacetic acid, 90% acetonitrile and 10% water as solvent for isocratic elution of the tracer and applied a TLC method using ITLC SG strips and the same solvent. The stability of the preparation was studied for 15 min, 3 h and 6 h. In order to evaluate the potential relevance of 131I-labeled methylene blue for melanoma detection, the in vitro uptake of 131I-methylene blue was investigated in SK-MEL 28 and 518A2 melanoma cells. Time and a temperature influence on uptake of 1311 methylene blue by these two melanoma cells were investigated. The radiochemical purity obtained by the HPLC method was 99.97 +/- 0.08% (n=8), while that by the TLC method was 99.88 +/- 0.16% (n=8). This indicates the excellent agreement between these two methods. The stability was persistent over 6 h and amounted to 99.75% +/- 0.21% (n=8). The uptake of 131I methylene blue was time and temperature dependent by both melanoma cells lines. The net cellular uptake on incubation at 37 degrees C of 131I methylene blue by SK-MEL 28 cells was high at 56.3-61.8% and that by 518A2-cells was 36.3-56.0%. Uptake by these cells was also investigated at 22 degrees C. The uptake by both cell types was also high at this temperature, but lower than that at 37 degrees C, amounting to 45.0-51.7% and 25.6-36.3%, respectively. Due to its easy handling and quite high uptake by melanoma cells, we expect that this tracer could be successfully used in routine application for melanoma imaging or eventual radiotherapy. PMID- 19189649 TI - Antiproliferative property and apoptotic effect of xanthorrhizol on MDA-MB-231 breast cancer cells. AB - Xanthorrhizol is a natural sesquiterpenoid compound isolated from the rhizome of Curcuma xanthorrhizza Roxb (Zingerberaceae). Recent studies of xanthorrhizol in cell cultures strongly support the role of xanthorrhizol as an antiproliferative agent. In our study, we tested the antiproliferative effect of xanthorrhizol using different breast cancer cell lines. The invasive breast cancer cell line, MDA-MB-231, was then selected for further investigations. Treatment with xanthorrhizol caused 50% growth inhibition on MDA-MB-231 cells at 8.67 +/- 0.79 microg/ml as determined by sulforhodamine B (SRB) assay. Hoechst 33258 nuclear staining assay showed the rate of apoptosis of MDA-MB-231 cells to increase in response to xanthorrhizol treatment. Immunofluorescence staining using antibody MitoCapture and fluorescein isothiocyanate (FITC)-labeled cytochrome c revealed the possibility of altered mitochondrial transmembrane potential and the release of cytochrome c respectively. This was further confirmed by Western-blotting, where cytochrome c was showed to migrate from mitochondrial fraction to the cytosol fraction of treated MDA-MB-231 cells. Caspase activity assay showed the involvement of caspase-3 and caspase-9, but not caspase-6 or caspase-8 in MDA-MB 231 apoptotic cell death. Subsequently, cleavage of PARP-1 protein is suggested. These data suggest treatment with xanthorrhizol modulates MDA-MB-231 cell apoptosis through the mitochondria-mediated pathway subsequent to the disruption of mitochondrial transmembrane potential, release of cytochrome c, activation of caspase-3 and caspase-9, and the modulation of PARP-1 protein. PMID- 19189651 TI - Association of polymorphisms of NOD2, TLR4 and CD14 genes with susceptibility to gastric mucosa-associated lymphoid tissue lymphoma. AB - BACKGROUND: The clinical course of Helicobacter pylori infection is highly variable and is influenced by both microbial and host factors, including the genetic composition of the infecting strains and variations in the host immune responses. A genetic risk profile for gastric cancer has been identified, but genetic susceptibility to develop gastric mucosa-associated lymphoid tissue (MALT) lymphoma is unclear. The aim of this study was to evaluate the relationship between NOD2, TLR4 and CD14 genetic polymorphisms and the development of gastric MALT-lymphoma. MATERIALS AND METHODS: Fifty-six patients with primary gastric MALT lymphoma and 51 patients with H. pylori infection were enrolled in this study. The polymorphisms were detected by the PCR-restriction fragment length polymorphism (RFLP) method of allele-specific PCR. RESULTS: No polymorphisms in the NOD2 and TLR4 genes were found to be associated with the development of gastric MALT lymphoma. Carriers of the CD14 gene -159T allele had a marginally increased risk of developing gastric MALT lymphoma than the controls (p=0.042). CONCLUSION: The -159C/T genetic polymorphism of the CD14 gene may be implicated in the development of gastric MALT lymphoma. PMID- 19189652 TI - Noscapine inhibits human prostate cancer progression and metastasis in a mouse model. AB - BACKGROUND: Noscapine, a non-toxic alkaloid and common constituent of cough medicine, stabilises tubulin. It inhibits the growth of several human and murine neoplasms, with no significant toxicity. Its effect on prostate cancer has not been evaluated. MATERIALS AND METHODS: Noscapine was administered orally (300 mg/kg per day) for 56 days to PC3 human prostate cancer-bearing immunodeficient mice (n=10). Immunodeficient control mice (n=10) received only diluent in an identical regimen. RESULTS: Mean total tumour weight was 0.42 +/- 0.23 g and 0.97 +/- 0.31 g (p<0.001) in the noscapine-treated group and the control group, respectively, without evidence of toxicity. Metastases occurred less frequently in the treatment than the control group (30% vs. 90%; p<0.05). CONCLUSION: Oral administration of noscapine limited tumour growth and lymphatic metastasis of PC3 human prostate cancer in this mouse model, supporting its therapeutic potential as a nontoxic and easily administered treatment for metastatic cancer. PMID- 19189653 TI - Zapotin prevents mouse skin tumorigenesis during the stages of initiation and promotion. AB - BACKGROUND: Zapotin, a flavonoid associated with Casimiroa edulis, was isolated as part of a program to discover natural inhibitors of carcinogenesis. Zapote blanco, the fruit of Casimiroa edulis, is consumed in many parts of the world. Zapotin is a non-toxic inducer of cellular differentiation, apoptosis and cell cycle arrest with cultured HL-60 promyelocytic cells. MATERIALS AND METHODS: An efficient chemical synthesis for zapotin was devised. Using this synthetic material, activity was examined in the two-stage mouse skin carcinogenesis model. RESULTS: Topical zapotin significantly inhibited 7,12 dimethylbenz(a)anthracene/12-O-tetradecanoylphorbol-13-acetate-induced mouse skin tumorigenesis, using the anti-initiation and anti-promotion protocols. CONCLUSION: Encouraging results from previous and current in vivo studies warrant further investigation of the chemopreventive activity of zapotin. PMID- 19189654 TI - Lack of involvement of the GNAS1 T393C polymorphism in prostate cancer risk in a Japanese population. AB - BACKGROUND: GNAS1 encodes the a-subunit of the Gs protein (Gsa), which binds GTP and stimulates adenylyl cyclase. Activating mutations lead to somatotroph, thyroid, adrenal and gonadal adenomas or the McCune-Albright syndrome and recently the T399C polymorphism in GNAS1 has been reported to be associated with malignancies. The purpose of the present case-control study with 349 Japanese prostate cancer patients and 203 urological controls was to determine whether the GNAS1 T393C polymorphism is associated with prostate cancer risk. MATERIALS AND METHODS: The GNAS1 T393C polymorphism was examined by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis. Odds ratios (OR) were adjusted for age using multiple logistic regression analysis with SPSS Medical Pack. RESULTS: The allele frequencies were compatible with the control population in Hardy-Weinberg equilibrium with 80, 169 and 100 for GNAS1 C/C, C/T and T/T, respectively in the patients with prostate cancer, compared with 42, 94 and 67 in the controls. No association between the GNAS1 polymorphism and prostate cancer risk was apparent. The C/C genotype was more frequent among the prostate cancer patients (22.9%) than the controls (20.7%), although without significance (OR, 1.30; 95% CI, 0.80-2.12; p=0.29). CONCLUSION: This pilot study does not support involvement of the GNAS1 polymorphism in prostate cancer risk. PMID- 19189655 TI - Omega-3 fatty acids suppress growth of SW620 human colon cancer xenografts in nude mice. AB - AIM: The purpose of this study was to examine the influence of fish oil on growth of colon cancer in nude mice. MATERIALS AND METHODS: Xenografts were initiated in mice receiving a standard diet or diets modified with corn or fish oil. After 3 weeks, mice were sacrificed, tumours were removed and processed for lipid analysis, histopathology and high resolution magic angle spinning magnetic resonance spectroscopy. RESULTS: Diet modified with fish oil suppressed tumour growth. Xenografts from mice receiving fish oil had higher levels of omega-3 polyunsaturated fatty acids (PUFAs) with concomitant reduced levels of omega-6 PUFAs. Furthermore, these xenografts had significantly lower levels of phosphocholine. Overall the results indicated less aggressive tumour growth in mice receiving a fish oil diet. PMID- 19189656 TI - Epidermal growth factor receptor pathway gene expressions and biological response of glioblastoma multiforme cell lines to erlotinib. AB - BACKGROUND: Erlotinib, an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor, exerts highly variable antiproliferative effects on human glioblastoma multiforme (GBM) cells in vitro and in vivo. As these effects are independent of EGFR baseline expression levels, more complex genetic signatures may form the molecular basis of the erlotinib-sensitive and erlotinib-resistant GBM phenotypes. The aim of the current study was to determine which genes within the EGFR signaling pathway are candidates for mediating the cellular response of human GBM towards erlotinib. MATERIALS AND METHODS: Complementary (c)RNAs from cell lines selected to represent the sensitive, intermediately responsive and resistant phenotypes, respectively, were hybridized to CodeLink Human Whole Genome Bioarrays. RESULTS: Expression analysis of the prospectively selected 244 genes whose products constitute the EGFR signaling pathway identified five genes the expression of which significantly correlated with phenotype. Functional annotation analysis revealed one (STATI) and two (FKBP14, RAC1) genes conclusively associated with sensitivity and resistance to erlotinib, respectively. Moreover, two additional genes (PTGER4, MYC) were unexpectedly found to be associated with sensitivity. The gene expressions were confirmed by quantitative polymerase chain reaction. CONCLUSION: Five genes within the EGFR signaling pathway may modulate GBM response to erlotinib, which further emphasizes the importance of this pathway for the biology of GBM. PMID- 19189657 TI - Pathogenetic pathways leading to glioblastoma multiforme: association between gene expressions and resistance to erlotinib. AB - BACKGROUND: The antiproliferative effects of erlotinib, an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor, on human glioblastoma multiforme (GBM) cell lines in vitro and in vivo are widely variable and independent of EGFR baseline expression levels, indicating that more complex genetic signatures may form the molecular basis of GBM response to erlotinib. This study sought to determine which genes within two common genetic pathways of GBM pathogenesis, i.e., the primary and secondary pathways, may be involved in mediating the cellular response of human GBM towards erlotinib. MATERIALS AND METHODS: Complementary (c)RNAs from cell lines selected to represent the sensitive, intermediately responsive and resistant phenotypes, respectively, were hybridized to CodeLink Human Whole Genome Bioarrays. RESULTS: Expression analysis of prospectively selected 104 genes pertaining to the primary and secondary pathways of GBM pathogenesis identified two genes (IGF1, PIK3C2B) the expression of which significantly correlated with cellular resistance towards erlotinib. CONCLUSION: Among the genes constituting two common pathways of GBM pathogenesis, two candidate genes may confer GBM resistance towards erlotinib, suggesting that resistance towards this compound may be acquired during the natural evolution of GBM. PMID- 19189658 TI - Genotypic determination by PCR-RFLP of human papillomavirus in normal oral mucosa, oral leukoplakia and oral squamous cell carcinoma samples in Madrid (Spain). AB - BACKGROUND: Human papillomaviruses (HPVs), especially genotypes 16 and 18, are considered to be human carcinogens by the International Agency for Research on Cancer (IARC). They are the most important etiological agents of uterine cervix cancer but their true role in oral carcinogenesis is controversial. OBJECTIVES: To detect the presence of HPV genome genotypes in oral leukoplakia and oral squamous cell carcinoma (OSCC) and analyze their relationship with clinicopathological variables. MATERIALS AND METHODS: Presence of genome ofHPV types 6, 11, 16, 18, 31, 33, 39, 42, 45, and 52 was studied by polymerase chain reaction in samples of normal mucosa (30 controls), oral leukoplakia (35 cases) and OSCC (33 cases). Results were compared between groups and differences were examined in relation to clinical and histological variables. RESULTS: HPV genome was detected in 23.3% of controls, 45.7% of oral leukoplakias, and 39.4% of OSCCs. Only HPV-16 was significantly (p=0.0005) more frequently detected in leukoplakias (40%) and OSCCs (33.3%) versus controls (0%). No significant relationship was found between the presence of viral genome and the main clinicopathological variables. CONCLUSION: According to these findings, the presence of HPV-16 is significantly associated with oral leukoplakia and OSCC lesions, therefore in our setting this virus may be a carcinogenic element in this disease. PMID- 19189659 TI - The endocannabinoid anandamide neither impairs in vitro T-cell function nor induces regulatory T-cell generation. AB - BACKGROUND: The cannabinoids have been proposed in the treatment of cancer. Generally, the cannabinoids are believed to be useful only in the palliative therapy of cancer-related symptoms, namely pain, anorexia and cachexia. However, preliminary experiments would also suggest an inhibitory effect of cannabinoids on cancer growth, whereas their influence on anticancer immunity is still controversial. The present study aimed to evaluate the influence of the endogenous cannabinoid anandamide (AEA) on T-cell phenotype and function. MATERIALS AND METHODS: The in vitro effects of AEA were evaluated at different concentrations on lymphocyte proliferation, cytotoxicity and differentiation, and in particular on T-regulator generation. RESULTS: AEA did not modify lymphocyte proliferation, neither under basal conditions, nor after IL-2 stimulation. Moreover, AEA did not induce the generation of regulatory T-lymphocytes nor the production of the immunosuppressive cytokine, IL-IO. CONCLUSION: The direct antitumor activity of AEA together with the absence of negative effects on T-cell functions might provide new insights into the potential use of cannabinoid agents in cancer immunotherapy. PMID- 19189660 TI - SUMO-1 overexpression increases RbAp46 protein stability and suppresses cell growth. AB - The retinoblastoma suppressor (Rb)-associated protein 46 (RbAp46) is a nuclear protein of 46 kDa and contains four repeats that end with Trp-Asp (WD) residues. In this study, we reveal that the RbAp46 protein level upon SUMO-1 expression was increased. The increasing level of RbAp46 protein by SUMO-1 was not regulated at the transcriptional level. SUMO-1 does not affect the degradation of RbAp46. Co localization of RbAp46 and SUMO-1 in the nuclei of stable NIH/3T3 cells harboring the inducible Ha-ras(Val12) oncogene (pSVlacOras) designated as 7-4, and protein protein interaction between RbAp46 and SUMO-1 was also detected by co immunoprecipitation in these cells. However, SUMO-l-related sumoylation was not involved in the modification of RbAp46. It is possibly that SUMO-1 acts through formation of complex with RbAp46 to stabilize RbAp46 protein. Overexpression of RbAp46 protein suppressed the NIH/3T3 cell growth induced by Ha-Ras(V12). SUMO-1 further enhances the suppression of cell growth through stabilization of RbAp46 protein. This is the first report to demonstrate that SUMO-1 can suppress Ras related cell proliferation through stabilization of RbAp46 protein. PMID- 19189661 TI - Inhibition of hepatocellular cancer by EB1089: in vitro and in vive study. AB - BACKGROUND: Despite significant antiproliferative potential, clinical application of 1,25 dihydroxy vitamin D3 [1,25(OH)2VD3] in the treatment of cancer has been hindered due to the development of hypercalcemia. Various derivatives of 1,25(OH)2VD3 have been synthesized to overcome this side-effect. Seocalcitol (EB1089) is a vitamin D analog that has been extensively studied and shown to have profoundly reduced hypercalcemic effects. Here the effects of EB1089 were evaluated in Hep 3B, SKHEP-1, PLC/PRF/5, HTC and Novikoff hepatocellular cancer (HCC) cell lines. MATERIALS AND METHODS: In vitro, cells were treated with different concentrations of EB1089 (1-1000 nM). Analytical tests were then performed including cell count and 3H thymidine assay. For in vivo analysis, SKHEP-1 cells were xenografted into nude male mice. Twenty-four hours after inoculation, mice were randomly assigned to a control group (n=10) or one of the treatment groups (3 groups of 10 mice) receiving 0.02, 0.1 or 0.5 microg/kg/day of EB1089. Control animals received the vehicle (propylene glycol). To minimize the number of intraperitoneal injections, oral and intraperitoneal routes were used on alternate days. Tumor size was measured every third day and the volumes were estimated using the formula 0.5 x length x (width)2. RESULTS: In vitro: Proliferation of Hep 3B, PLC/PRF/5 and SKHEP-1 HCC cells was significantly inhibited at all EB1089 concentrations tested, while HTC cells only responded to 1000 nM concentration of EB1089. Proliferation of Novikoff cells was unaffected by the drug at all concentrations examined. In vivo: EB1089 effectively inhibited SKHEP-1 tumor growth without inducing hypercalcemia (p<0.05). CONCLUSION: Results of the present study indicate that EB1089 is an effective growth inhibitor of HCC tumors. PMID- 19189662 TI - Expression of Mcm7 and Cdc6 in oral squamous cell carcinoma and precancerous lesions. AB - BACKGROUND: The present study aimed at evaluating the clinical importance of Mcm7 and Cdc6 expression in oral squamous cell carcinoma (OSCC) and precancerous lesions. MATERIALS AND METHODS: RT-PCR and immunohistochemistry analysis were performed on 47 frozen samples and 98 paraffin-embedded samples to evaluate the mRNA and protein expressions of Mcm7 and Cdc6. RESULTS: RT-PCR and immunohistochemistry indicated positive expressions of Mcm7 mRNA and protein in normal oral mucosa, precancerous lesions and OSCC. Significant differences were found between all the groups. Cdc6 mRNA and protein had low expressions in normal oral mucosa but were highly expressed in precancerous lesions and OSCC. Mcm7 and Cdc6 expressions in the lymph node metastasis cases were significantly higher than those of the nonmetastatic carcinomas. CONCLUSION: High expressions of Mcm7 and Cdc6 are correlated with the development and metastasis of OSCC and may become a molecular marker for the early diagnosis and prognosis prediction for OSCC. PMID- 19189663 TI - Correspondence between flat aberrant crypt foci and mucin-depleted foci in rodent colon carcinogenesis. AB - BACKGROUND: Flat aberrant crypt foci (flat ACF) and mucin-depleted foci (MDF) are preneoplastic lesions identified in the colon of carcinogen-treated rodents stained with methylene blue (MB) and high iron diamine-alcian blue (HID-AB), respectively. The correspondence between flat ACF and MDF in the same colon of Min mice treated with azoxymethane (AOM) and of F344 rats treated with 1,2 dimethylhydrazine (DMH) was explored. MATERIALS AND METHODS: The position of each flat ACF was recorded on a digitally constructed photographic map of the MB stained colon. The same colons were then stained with HID-AB and the position of each MDF was compared with that of flat ACF. RESULTS: The fraction of coincident lesions, identified as both flat ACF and MDF with the two staining methods, was 57% and 42%, in the Min mice and F344 rats, respectively. Flat ACF or MDF not coincident with the two staining methods were either undetectable or ACF with one of the two methods. CONCLUSION: Flat ACF and MDF show considerable, but not total, overlap. PMID- 19189664 TI - Berberine inhibits human neuroblastoma cell growth through induction of p53 dependent apoptosis. AB - Berberine, an alkaloid, has anti-tumor properties in some cancer cells, but action mechanisms are not clear yet. We here investigated the anticancer activity of berberine and possible mechanisms in human neuroblastoma SK-N-SH and SK-N-MC cells. The p53-expressing cells, SK-N-SH (IC50=37 microM) were more susceptible to berberine than the p53-deficient cells, SK-N-MC (IC50 > or =100 microM) without cytotoxic effect on the cortical neuronal cells. Berberine caused cell cycle arrest in G0/G1 phase and apoptotic cell death, and these effects were much greater in SK-N-SH cells than those in SK-N-MC cells. Berberine much greatly decreased G0/G1 phase-associated cyclin and cyclin-dependent kinase (cyclin D1, cyclin E, Cdk2, and Cdk4) expression, and increased apoptotic gene expression and activation of caspase-3 in SK-N-SH cells. Exploration of p53 siRNA or pifithrin alpha (PFT-alpha), a p53 inhibitor, in the SK-N-SH cells resulted in increase of IC50 values for cell viability, and decreased apoptotic cell death, expression of p53 and activation of caspase-3. Therefore, these results showed that berberine causes p53-dependent apoptotic death of neuroblastoma cells, and suggested that berberine may be useful as an anticancer agent for neuroblastoma. PMID- 19189665 TI - Effects of 5-fluorouracil on human mitogen-activated peripheral blood lymphocytes from healthy individuals. AB - BACKGROUND: The effect of 5-fluorouracil (5-FU) on activated lymphocytes was explored. MATERIALS AND METHODS: The in vitro effects of 5-FU on DNA synthesis in mitogen-activated lymphocytes from healthy volunteers were compared to those of the antimetabolites doxorubicin, cyclophosphamide and 6-mercaptopurine. These effects were assessed by alterations in the phenotypic profile and the percentage of cells in various phases of the cell cycle, as well as by the secretion of T helper (Th)1 and Th2 cytokines (ELISA). RESULTS: Unlike 5-FU, the other antimetabolites failed to augment DNA synthesis in activated lymphocytes. The effect of 5-FU correlated with an increase in the percentage of cells in the S phase and caused an increased in CD4+ cells, a decrease in CD56+ cells and a shift of the cytokine secretion pattern from Th2 to Th1. CONCLUSION: 5-FU exhibited a unique effect on DNA synthesis in activated lymphocytes which was accompanied by selective effects on various lymphocyte subpopulations. PMID- 19189666 TI - p53, p16 and Fhit proteins expressions in chronic esophagitis and Chagas disease. AB - BACKGROUND: Models have suggested esophageal carcinogenesis can result from the alteration of sequences, leading to esophagitis, atrophy, dysplasia, carcinoma in situ and invasive carcinoma. While numerous genetic alterations have been reported in esophageal carcinogenesis, studies of benign lesions with precancerous potential are scarce. MATERIALS AND METHODS: Immunohistochemistry was performed for p53, p16 and Fhit proteins in the esophageal mucosa from patients with Chagas disease (CD), chagasic megaesophagus (CM), chronic esophagitis (CE), esophageal squamous cell carcinoma (ESCC) and in normal mucosa (NM). RESULTS: The proportion of p53-positive cases increased progressively according to the severity of the pathology CD (7.7%), CM (26.1%), CE (522%) and ESCC (100%). However, p16 and Fhit did not show any statistically significant differences among the groups. CONCLUSION: p53 overexpression is involved in the initial steps of esophageal carcinogenesis, supporting further evaluation of its utility as a marker in precursor lesions, conversely, losses of Fhit and p16 expression may not be significant. PMID- 19189667 TI - mTOR inhibition sensitizes gastric cancer to alkylating chemotherapy in vivo. AB - BACKGROUND: Gastric cancer is a highly chemoresistant tumor. Previous studies suggest that cancer cells can be sensitized to standard chemotherapy, and especially alkylating agents, by inhibition of mammalian target of rapamycin (mTOR) signaling. The work presented here shows that the mTOR inhibitor everolimus, in combination with cyclophosphamide, exhibits synergistic antitumor activity in gastric cancer xenografts. MATERIALS AND METHODS: Treatment with everolimus at the minimal effective dose was studied in combination with cyclophosphamide at maximum tolerated dose in a human gastric cancer severe combined immunodeficient (SCID) mouse xenograft model. Besides tumor size, biomarker expression for proliferation (Ki-67), hypoxia (HIF-12alpha), apoptosis (activated caspase 3), angiogenesis (microvascular density, MVD) and levels of circulating endothelial progenitors (CEPs) were assessed. RESULTS: Everolimus single agent treatment significantly inhibited tumor growth relative to control and cyclophosphamide treatment (T/C 19%, p<0.01). This antitumor activity was linked to a significant decrease in tumor cell proliferation (p<0.01) and a trend towards lower tumor MVD, HIF-1alpha expression and levels of CEPs. Notably, the combination of everolimus with cyclophosphamide resulted in synergistic anti tumor activity (T/C 9%, p<0.01). This antitumor activity coincided with a statistically significant decrease in MVD (p<0.01). Whereas treatment with everolimus was well tolerated, cyclophosphamide at maximum tolerated dose (MTD) showed significant toxicity both as monotherapy and in combination with everolimus. CONCLUSION: The antiangiogenic activity of everolimus combined with a high dose of cyclophosphamide shows synergistic antitumor activity against gastric cancer in vivo. In potential future clinical trials, the toxicity of cyclophosphamide in combination regimens with everolimus deserves careful evaluation. PMID- 19189668 TI - A novel approach for gene transduction with adenovirus vector and the fibrin glue system. AB - BACKGROUND: The fibrin glue system (FGS) consists of liquid forms of fibrinogen and thrombin and is used widely in surgery for hemostasis. In this study, as a novel and unique approach, the possibility and efficacy of locoregional gene transfer using the FGS containing an adenoviral vector was determined. MATERIALS AND METHODS: The optimum concentration of the adenoviral vector mixed with the FGS (AxCALacZ/FGS) for gene transduction was evaluated in vitro by X-gal (beta galactosidase) staining and NIH (National Institute of Health) imaging in RCN-9, a rat colon carcinoma cell line. To determine the survival period of the adenoviral vector in the fibrin glue, RCN-9 cells were exposed to AxCALacZ/FGS after it had been incubated for various periods and the transduction efficiencies were evaluated by beta-galactosidase (beta-gal) assay. AxCALacZ/FGS was also applied in vivo to the resected site of rat liver. AxCALacZ diluted in PBS (AxCALacZ/PBS) was used as the control. The transduction efficiencies in the liver were compared by X-gal staining and beta-gal assay. RESULTS: Almost 100% transgene expression was demonstrated by the X-gal staining and NIH imaging at a concentration level greater than 1 multiplicity of infection. LacZ expression (as beta-galactosidase) revealed gene-transduced RCN-9 cells when the AxCALacZ/FGS was held for a period of less than 96 hours. The treatment with the AxCALacZ/FGS in vivo resulted in greater transgene expression than the treatment with AxCALacZ/PBS. CONCLUSION: The adenoviral vector survives and remains stable in the FGS for sufficient time for transduction to occur and AxCALacZ/FGS can efficiently transduce the target gene both in vitro and vivo. PMID- 19189669 TI - The E-cadherin adhesion molecule and colorectal cancer. A global literature approach. AB - The E-cadherin-catenin complex plays a crucial role in epithelial cell cell adhesion and in the maintenance of tissue architecture. Down-regulation of E cadherin expression correlates with a strong invasive potential, resulting in poor prognosis in human carcinomas. Progress has been made in understanding the interaction between the different components of this protein complex and how this cell-cell adhesion complex is modulated in cancer cells. The present study is an update of the role of E-cadherin in human colorectal cancer. It emphasizes new features and the possible role of the complex in clinical practice, discussed in the light of references obtained from the Medline database from 1987 to 2007. In colorectal carcinomas, changes in E-cadherin expression have been correlated with tumour size, histopathology and differentiation, but results are still inconsistent. Further studies may yield greater insight into the early molecular interactions critical to the interaction between adhesion molecules and tumour initiation and progression. This should aid the development of novel strategies for both prevention and treatment of cancer. PMID- 19189670 TI - p38alpha-selective MAP kinase inhibitor reduces tumor growth in mouse xenograft models of multiple myeloma. AB - Multiple myeloma (MM) is a clonal plasma cell malignancy, which is currently incurable. Therefore, new mono- or combined therapy treatment regimens in the early and advanced phases of MM are urgently needed to combat this disease. Recently, p38 mitogen-activated protein kinase (MAPK) has been implicated as playing an important role in MM. Therefore, the effect of a p38alpha-selective MAPK inhibitor, SCIO-469 (indole-5-carboxamide, ATP-competitive inhibitor), or its structural analog, SD-282 (indole-5-carboxamide, ATP-competitive inhibitor) was examined in mouse xenograft models of MM using human RPMI-8226 or H-929 plasmacytoma inocula. Oral treatment with SCIO-469 (10, 30, 90 mg/kg) twice daily was initiated in mice with palpable tumors of RPMI-8226 origin, a condition that mimics early human myeloma disease. In mice with palpable tumors, 14 days of SCIO 469 treatment significantly reduced RPMI-8226 tumor growth in a dose-dependent manner. A significant dose-dependent reduction in RPMI-8226 tumor growth was also observed when SCIO-469 oral treatment at doses of 10, 30 and 90 mg/kg twice daily was initiated in mice with tumors of pronounced size, a condition that mimics advanced human myeloma disease. In a similar set of studies employing the SCIO 469 analogue SD-282 at 90 mg/kg/bid orally, histological assessment at the end of the study demonstrated a significant reduction in RPMI-8226 tumor growth and angiogenesis. SD-282 treatment was additionally shown to significantly reduced expression of heat-shock protein-27 (HSP-27) and phospho-p38 in the tumor cells. Furthermore, co-administration of SCIO-469 with dexamethasone elicited antitumor properties in dexamethasone-sensitive H-929 tumors at much lower than the typically effective doses of dexamethasone, suggesting its potential for combined therapy. In conclusion, p38 inhibitors reduced human myeloma cell growth in vivo both at early and advanced phases of the disease. The current study also provides evidence of potential for co-therapy with dexamethasone. PMID- 19189671 TI - Adhesion of a two-step etch-and-rinse adhesive on collagen-depleted dentin. AB - PURPOSE: To evaluate microtensile bond strength and interfacial nanoleakage expression of adhesive interfaces created by XP-Bond on human deproteinized dentin immediately after bonding and after 6 months of artificial aging. MATERIALS AND METHODS: Noncarious human molars were selected, middle/deep dentin substrates were exposed, and either assigned to group 1 (XP-Bond applied on collagen-depleted dentin) or to group 2 (XP-Bond applied in accordance with manufacturers' instructions). In group 1, the etched dentin surface was treated with 10% NaOCl for 60 s to remove the exposed demineralized organic matrix before XP-Bond application. Composite/dentin beams were obtained in accordance with the microtensile nontrimming technique and either pulled to failure after 24 h or after 6 months' artificial aging. Data were analyzed with two-way ANOVA and Tukey's post-hoc test (p < 0.05). Interfacial nanoleakage evaluation was performed on additional adhesive interfaces to quantify the amount of silver tracer along the interface. RESULTS: The use of NaOCI before XP-Bond application (group 1: 18.9 +/- 5.8 MPa) reduced immediate bond strength by 62% compared to controls (group 2: 49.9 +/- 10.3 MPa; p < 0.5). After 6 months of artificial aging, the bond strength of groups 1 and 2 significantly decreased to 10.1 +/- 2.7 MPa and 35.2 +/- 8.7 MPa, resp (p < 0.05). Interfacial nanoleakage expression along XP-Bond adhesive interfaces was increased either by sodium hypochlorite or by artificial aging. CONCLUSION: The role of collagen fibrils is pivotal for the bonding of XP-Bond to dentin, as decreased immediate bond strength and reduced bond stability over time was found on collagen-depleted dentin. PMID- 19189672 TI - Comparison of bonding performance of self-etching and etch-and-rinse adhesives on human dentin using reliability analysis. AB - PURPOSE: To estimate the in vitro reliability of typical self-etching and etch and-rinse adhesives of various application protocols. MATERIALS AND METHODS: The following adhesives were applied on flat dentin surfaces of extracted human teeth (n = 223): self-etching two-step adhesives: AdheSE (AH), Clearfil SE Bond (CL), OptiBond SE (OS); one-step adhesives: Adper Prompt L-Pop (ADP), Adper Prompt (AD), and Xeno III (XE); all-in-one adhesive: iBond (IB); etch-and-rinse three step adhesives: OptiBond FL (OF), two-step Gluma Comfort Bond (G), Excite (E) and Prime & Bond NT (PB). Composite buildups were prepared using a microhybrid composite, Opticor New. Shear bond strength was determined after 24 h of storage at 37 degrees C in distilled water. The results were analyzed with a nested ANOVA (adhesive, type of adhesive) followed by the Fisher post-hoc tests of group homogeneity at alpha = 0.05. A two-parameter Weibull distribution was used to calculate the critical shear bond strength corresponding to 5% probability of failure as a measure of system reliability. RESULTS: ANOVA revealed a significant decrease (p < 0.001) in the mean shear bond strength as follows: AH=CL=OS=G=E=OF>AD=IB=XE>PB=ADP, but no significant difference (p > 0.48) between the etch-and-rinse and self-etching adhesives. The corresponding characteristic bond strength of Weibull distribution ranged between 24.1 and 12.1 MPa, Weibull modulus between 8.3 and 2.1, and the critical shear bond strength varied from 16.0 to 3.0 MPa. CONCLUSION: Pronounced differences in the critical shear bond strength suggest reliability variations in the adhesive systems tested, which originate from chemical composition rather than type of adhesive. PMID- 19189673 TI - Effect of caries-affected dentin hardness on the shear bond strength of current adhesives. AB - PURPOSE: The aim of this study was to evaluate the microshear bond strength of three current adhesives to normal and caries-affected dentin, and to examine the correlation between dentin nanoindentation hardness and bond strength. MATERIALS AND METHODS: Nanoindentation hardness of dentin and microshear bond strength of Clearfil SE Bond, Clerafil Tri-S Bond (Kuraray Medical) and Single Bond (3M ESPE) were measured on caries-affected and normal dentin. Modes of fracture for the bonding tests and the resin/dentin interfaces were observed using SEM. Data were analyzed at a statistical significance level of p < 0.05. RESULTS: Significantly lower hardness values were recorded for caries-affected dentin compared to normal dentin. Two-way ANOVA showed that bonding material, type of dentin, and the interactions of these two factors had significant effects on bond strength. A significant correlation was found between dentin hardness and microshear bond strength. However, ANCOVA analysis revealed that the effect of dentin type was not only due to the mechanical properties of dentin. One-way ANOVA with Tukey's HSD test revealed that the mean bond strength of normal dentin with Tri-S Bond was significantly lower than that with SE Bond and Single Bond; however, bond strengths to caries-affected dentin showed no significant difference between the three adhesives. CONCLUSION: All three adhesives showed lower bond strength to caries-affected dentin than to normal dentin. The differences between adhesives were not significant with caries-affected dentin as the bonding substrate. Dentin mechanical properties are not the only factors responsible for lower bond strength to caries-affected dentin. PMID- 19189674 TI - Effect of ultrasonic agitation on bond strength of self-etching adhesives to dentin. AB - PURPOSE: The aim of this study was to evaluate the influence of ultrasonic treatment on the microtensile bond strength of self-etching adhesives to dentin. MATERIALS AND METHODS: Forty-two human molars were wet ground occlusally until dentin was exposed. Clearfil S3 Bond, Futurabond NR (one-bottle) and Clearfil SE Bond (two-bottle) self-etching bonding systems were used in this study. In control groups, bonding procedures were performed according to the manufacturers' instructions. In the experimental groups, bonding materials were applied with an ultrasonic scaler. When using Clearfil SE Bond, the ultrasonic device was used either during priming or the bonding stage. The composite was then built up to 5 mm in height. Each tooth was serially sectioned into rectangular beams. The beams were categorized also according to positional status as marginal or central. Beams were subjected to microtensile testing after 24 h of water storage. Failure modes were observed with a stereomicroscope and classified. Randomly selected tested beams from each group were examined with SEM. RESULTS: Three-factor ANOVA results indicated that the adhesive bonding system had a significant effect on bond strength (p < 0.001), whereas ultrasonic agitation and the position of the tested beam (marginal vs central) had no effect on bond strength (p > 0.05). Failure after the test was commonly due to adhesive breakdown associated with partial cohesive failure in the dentin. The mean (SD) microtensile bond strengths to dentin for S3 Bond, Futurabond NR, Clearfil SE Bond in the control group were 44.3 (11.7), 35.3 (12.0), 25.1 (8.8), resp, and in the ultrasonic group 39.3 (14.2), 31.3 (13.5), 35.5 (13.5) at priming and 32.6 (16.2) at bonding. CONCLUSION: Ultrasonic agitation during application of self-etching adhesives had no effect on bonding performance of the self-etching adhesive. PMID- 19189675 TI - The effects of cavity size and filling method on the bonding to Class I cavities. AB - PURPOSE: The effects of incremental filling technique and flowable liner on the bond strength of resin composite in different sized cavities were investigated in this study. MATERIALS AND METHODS: Two sizes of cylindrical Class I cavities were prepared in bovine dentin (98.1 mm3 and 21.2 mm3). A light-curing resin composite (Clearfil Photocore) with a self-etching primer adhesive (Clearfil SE Bond) was used to bulk fill the cavities (C-factor 5), with and without flowable liner (Clearfil FLOW-FX) or incremental filling (2 layers, C-factor 3). As control, a 3 mm or 5-mm buildup was placed on the flat dentin surface (C-factor 0.2). After 24 h storage in 37 degrees C water, the microtensile bond strength was measured at a crosshead speed of 1 mm/min. Mode of failure in the fractured specimens was observed using a scanning electron microscope. The obtained results were statistically analyzed. RESULTS: In large cavities, incrementally filled and lined cavities showed higher bond strength values than the bulk-filled subgroup (p < 0.05). However, in small cavities, neither the incremental technique nor flowable liner improved the bond strength. CONCLUSION: Not only the filling techniques but also the cavity size affected the bond strength to the cavity floor in cylindrical Class I cavities. PMID- 19189676 TI - Does adhesive resin application contribute to resin bond durability on etched and silanized feldspathic ceramic? AB - PURPOSE: To assess the effect of adhesive application and aging on the bond durability of resin cement to etched and silanized feldspathic ceramic. MATERIALS AND METHODS: Twenty blocks (6.4 x 6.4 x 4.8 mm) of feldspathic ceramic (Vita VM7) were produced. The ceramic surfaces were conditioned with 10% hydrofluoric acid gel for 60 s and silanized. They were then randomly divided into two groups. While half of the group received no adhesive, in the other half, a layer of adhesive (Scotchbond Multi-Purpose Plus) was applied. Each ceramic block was then placed in its silicone mold with the treated surface exposed. The dual-cured resin cement (Variolink II) was injected into the mold over the treated surface and polymerized. Specimens were sectioned to achieve nontrimmed bar specimens (approximately 12 sticks/block) that were randomly divided into 2 groups: a) non aged--microtensile bond test immediately after sectioning; b) aged-thermocycling (TC) 12,000 times, 5 degrees C to 55 degrees C, and water storage (50 days). The microtensile bond strength test was performed in a universal testing machine (crosshead speed: 1 mm/min). The failure types were examined using an optical light microscope and SEM. Bond strength results were analyzed using two-way ANOVA and Tukey's test (alpha = 0.05). RESULTS: The adhesive application affected the bond strength results significantly (p = 0.0001) (without adhesive > with adhesive). While aging conditions did not reduce the bond strength in the groups that received no adhesive (20 +/- 5.3 MPa non-aged and 21.5 +/- 5.6 aged) (p = 0.1698), it significantly affected the bond strength results of the group with adhesive application (18 +/- 4.4 MPa to 14.4 +/- 4.7 MPa) (p < 0.001). All groups showed mainly mixed type of failures between the ceramic and the resin cement (81% to 100%). The group in which no adhesive was applied presented a higher incidence of cohesive failure of ceramic after aging (18%) than those of the other groups. CONCLUSION: The use of adhesive did not improve resin cement adhesion to the etched and silanized feldspathic ceramic after long-term thermocycling and water storage. PMID- 19189677 TI - In vitro evaluation of failure loads of nonmetal cantilevered resin-bonded fixed dental prostheses. AB - PURPOSE: To evaluate in vitro the influence of fiber reinforcement on the failure loads of resin composite beams, simulating cantilevered two-unit resin-bonded fixed dental prostheses, and compare the results with similarly obtained failure loads of ZrO2 and CoCr beams of a comparable design. MATERIALS AND METHODS: Peel tests were performed using resin composite, fiber-reinforced resin composite, and zirconia beams, simulating two-unit cantilevered resin-bonded fixed dental prostheses, luted with Panavia F2.0 onto flat-ground buccal surfaces of bovine mandibular incisors. The recorded failure loads were compared with those of CoCr beams of a similar size and design from earlier research. Finite element analysis revealed the stress concentrations within the cement layers at failure. RESULTS: The failure loads (N) of the peel tests, depending on the beam type and including the type of failure, were statistically analyzed. The highest failure values were obtained with the fiber-reinforced resin composite beams, which were luted with the exposed fibers directly on the bovine enamel. Finite element analysis showed that peak stress locations depend on the beam type and facilitate the explanation of the different failure modes. CONCLUSION: Fiber-reinforcement of simulated two unit cantilevered resin composite resin-bonded fixed dental prostheses does not necessarily lead to higher failure loads. This study identified significant differences in peel failure loads between identical specimens, depending on whether or not the fiber reinforcement was exposed on the luting surface. Further research needs to be carried out regarding the combination of resin composite and fiber reinforcement. PMID- 19189678 TI - Shear bond strength of glass-ionomer adhesive to dentin: effect of smear layer thickness and different dentin conditioners. AB - PURPOSE: To investigate the effect of two different smear layer thicknesses treated with different dentin conditioners on the bond strength of glass-ionomer adhesive to dentin. MATERIALS AND METHODS: Fifty human molars were used. Each tooth was sectioned in a mesiodistal direction into halves, using a low-speed diamond disk. The 100 halves were randomly divided into 10 equal groups. Groups I to V for the thick smear layer and groups VI to X for the thin smear layer. Eighty halves were used for shear bond strength testing, while the remaining 20 halves were used for SEM evaluation of the adhesive/dentin interface. After each dentin pretreatment, the glass-ionomer adhesive was applied over the conditioned dentin surfaces for all tested groups and restored with resin composite cylinders. Each half with its attached composite cylinder was subjected to shear load at crosshead speed of 0.5 mm/min until failure occurred. The dentin surfaces for SEM evaluation were treated as described for shear bond strength testing. The samples were examined using SEM operated at 30 kv, and the resin/dentin interface was examined at 1500X. RESULTS: Two-way ANOVA showed that there was no significant effect of the smear layer thickness on shear bond strength. There was a significant effect of the different conditioners used (p = 0.001). The interaction of the two independent variables (smear layer thickness and the dentin conditioners) showed no significant difference in the shear bond strength to dentin. The common feature in all evaluated specimen interfaces was the presence of a hybrid layer of different thicknesses with no visible resin tags. CONCLUSIONS: The shear bond strength of the glass-ionomer adhesive to dentin proved to be independent of the smear layer thickness. The use of dentin conditioner prior to the application of the glass-ionomer adhesive appeared to be an important step to improve its bond strength to dentin. Glass-ionomer adhesive failed to produce resin tags regardless of the type of dentin conditioner used. PMID- 19189679 TI - Effects of pretreatment and thermocycling on bond strength of resin core materials to various fiber-reinforced composite posts. AB - PURPOSE: To evaluate the effects of pretreatment (silanization) and thermocycling on bond strengths of 2 core materials to 3 different types of fiber posts. MATERIALS AND METHODS: Bond strengths of prefabricated glass (group 1), quartz (group 2), and individually formed glass fiber posts (group 3) to either a flowable or a highly viscous composite resin core material were measured using a push-out design. All posts were investigated with or without the application of a silane. Half of the specimens were stored in distilled water for 28 days, and were subsequently subjected to thermocycling (5000 cycles, 5 to 55 degrees C), whereas the other half was investigated after 24-h water storage. RESULTS: Bond strengths were significantly affected by thermocycling (p < 0.0005), post type (p < 0.0005), and pretreatment (p < 0.0005), but not by the core material (p = 0.149; four-way ANOVA). Posts of group 3 demonstrated significantly higher bond strengths than all other posts, even after thermocycling (p < 0.05; Tukey-B). CONCLUSION: While silanization had negligible effects, thermocycling drastically reduced bond strengths regardless of pretreatment, post type, or core material. The higher bond strengths between the group 3 posts and the flowable composite resin core material might result in improved stability of the core for postendodontic restorations. PMID- 19189680 TI - How should we diagnose and monitor central precocious puberty? PMID- 19189681 TI - Growth hormone receptor deficiency in South America: colonial history, molecular biology, and growth and metabolic insights. PMID- 19189682 TI - Congenital anodontia in ectodermal dysplasia. PMID- 19189683 TI - The diagnostic value of a brief GnRH analogue stimulation test in girls with central precocious puberty: a single 30-minute post-stimulation LH sample is adequate. AB - To ascertain the diagnostic value of GnRHa stimulation testing in girls with CPP, single sample 30 minute post-stimulated gonadotropin levels were compared between girls with CPP and prepubertal girls. Serum LH and FSH concentrations were assayed using two third generation gonadotropin assays. Clinical data were reviewed to establish the diagnosis of CPP. GnRHa stimulation testing with one LH measurement obtained 30 minutes after stimulation is adequate for evaluating girls with CPP and as reliable as GnRH stimulation testing. PMID- 19189684 TI - E180splice mutation in the growth hormone receptor gene in a Chilean family with growth hormone insensitivity: a probable common Mediterranean ancestor. AB - Mutations in the GH receptor gene have been identified as the cause of growth hormone insensitivity syndrome (GHIS), a rare autosomal recessive disorder. We studied the clinical and biochemical characteristics and the coding sequence and intron-exon boundaries of the GH receptor gene in a consanguineous family with severe short stature which consisted of two patients, their parents and five siblings. The two adolescents had heights of -4.7 and -5.5 SDS, respectively, with elevated growth hormone associated with low IGF-I, IGFBP-3 and GHBP concentrations. Molecular analysis of the GH receptor gene revealed a mutation in exon 6, present in both patients This mutation, E180 splice, has been previously described in an Ecuadorian cohort, and in one Israeli and six Brazilian patients. We determined the GH receptor haplotypes based on six polymorphic sites in intron 9. Co-segregation of the E180splice mutation with haplotype I was found in this family, compatible with a common Mediterranean ancestor, as shown for previous cases with the E180splice mutation described to date. PMID- 19189685 TI - Reliability of the low dose synacthen test in children undergoing pituitary function testing. AB - BACKGROUND: There are few data in the paediatric literature on the normal cortisol response to stimulation during the low dose synacthen test (LDST) (1 microg). AIM: To examine the cortisol responses in children, subsequently presumed to be normal, who had an LDST during anterior pituitary function tests (APFTs). METHODS: A retrospective review of results in children with short stature and normal growth hormone levels. RESULTS: Of 33 children tested, seven had suboptimal cortisol responses based on accepted criteria (peak <500 nmol/l)- a false positive rate of 21%. Only three of these children had a repeat LDST, which was normal in all cases. The peak cortisol response (median 633, range, 417 1052 nmol/l) was inversely correlated with age (r = -0.44, p < 0.05). CONCLUSION: One in five tests did not meet normal criteria. This false positive rate (21%) should be borne in mind when interpreting synacthen tests to prevent overdiagnosis of adrenal insufficiency. PMID- 19189686 TI - The leptin/adiponectin ratio in mid-infancy correlates with weight gain in healthy term infants, but is unrelated to serum insulin concentrations, body mass index, or skin fold thickness. AB - AIM: Rapid growth in infancy may be associated with later onset childhood obesity. The aim in this study was to evaluate the relationships of adipokines to growth of infants at 6-10 months of age and to serum insulin, glucose, and auxological parameters of infants and their mothers. METHODS: Thirty-seven healthy term AGA formula fed infants, 21 males, mean age 7.0 +/- 1.2 (SD) months, were evaluated during a nutritional assessment at a county health department. Length, weight, head circumference, waist circumference, mid-arm circumference, and subscapular skin fold and triceps skin fold thickness were determined. Mothers were weighed and their height measured, birth weight recorded from clinic records, and the infant's dietary history reviewed. Following finger stick for assessment of hemoglobin, a bedside blood glucose was determined and 250 microl of additional serum taken for assay of total adiponectin, high molecular weight (HMW) adiponectin (n=12), leptin, and insulin. RESULTS: The infants' total adiponectin to leptin ratio correlated significantly with the change in body weight from birth to mid-infancy (r = 0.349, p < 0.05). The mean total adiponectin was 34.2 - 16.6 microg/ml (n=37), mean HMW adiponectin 12.2 +/- 9.0 microg/ml (n=12), mean HMW/total adiponectin ratio 34.3 +/- 17.6%, and mean leptin 1.3 +/- 1.2 ng/ml. Neither total nor HMW adiponectin, leptin, nor the leptin/adiponectin ratio, correlated with serum insulin, glucose/insulin ratio, hemoglobin, birth weight, or auxological determinations of the infants or mothers. CONCLUSIONS: As leptin and adiponectin are both insulin sensitizing hormones that change inversely with acquisition of body fat, and the leptin/adiponectin ratio correlates significantly with weight gain in mid infancy, we postulate that this ratio might provide a marker relating to infantile growth and later adiposity. PMID- 19189687 TI - Identification of sequence variants in the UBL5 (ubiquitin-like 5 or BEACON) gene in obese children by PCR-SSCP: no evidence for association with obesity. AB - BACKGROUND: Childhood obesity has a strong genetic background. The human UBL5 (BEACON) gene has been suggested as a candidate gene for obesity. Previous studies in populations of different ethnicities have shown a significant association between UBL5 variants and measures of body fatness. AIMS: To identify mutations that may cause early-onset obesity we screened the UBL5 gene for sequence variations in a cohort of obese children who also had at least one obese parent (BMI >30 kg/m2) diagnosed before the age of 30 years. METHODS: We screened the UBL5 gene by PCR-SSCP and sequencing in a cohort (n=30) of obese children (mean age 6.9 +/- 3 yr), and then analysed SNPs by HRMA in a population of 160 obese and 140 lean individuals. RESULTS: Three sequence variations were detected: -422T>C in the 5'-UTR region, and -800T>A (rs10418248) and -860G>T in the promoter region. The SNPs -422 T>C in the 5'-UTR region and -860G>T have never been described before. These two SNPs did not co-segregate with obesity in relatives of the obese carriers. However, since in silico analysis of the -860G>T SNP region predicted a loss of the consensus binding site for RXR-alpha and RXR beta, both involved in adipose cell regulation, we screened the -860G>T variant in a cohort of 300 individuals, 160 young obese (mean age 33 years) and 140 lean individuals. No differences in genotype distribution or in -860T allele frequencies were found between the two groups (1.8% vs. 1.4%, p = NS). In addition, no association was found between obesity and the previously described 800T>A SNP (rs10418248). CONCLUSION: Our data suggest that the UBL5 gene is unlikely to play a major role in the genetic susceptibility to early-onset obesity in our population. PMID- 19189688 TI - Characteristics of primary hyperparathyroidism in adolescents. AB - AIM: To analyze adolescent primary hyperparathyroidism (PHPT) in India. DESIGN: A retrospective analysis of consecutive medical records of patients with PHPT < or =20 years old who underwent parathyroid surgery at a tertiary care center in north India from 1993 to 2006. METHODS: Fourteen adolescents underwent clinical, biochemical, hormonal, radiological and scintigraphic evaluation for confirmation of PHPT followed by parathyroidectomy. RESULTS: Mean (+/- SD) age of these patients was 15 +/- 3.9 years (8 female). All were symptomatic at presentation except one who had multiglandular involvement as a part of multiple endocrine neoplasia (MEN)-I screening. The common presenting manifestations included bone disease, recurrent nephrolithiasis and pancreatitis. Hypercalcemia was observed in eight of these patients. The mean (+/- SD) serum intact parathyroid hormone was 781 +/- 586 pg/ml. Ultrasonography localized an abnormal parathyroid gland in 12 (92%) of 14 patients, while 99mTc sestamibi scan (performed in 7) localized abnormal parathyroid glands in all (100%). All patients underwent bilateral neck exploration except one with MEN-I, who refused surgery. Twelve patients had single parathyroid adenoma and one had four-gland hyperplasia. Mean follow up was 44.1 months and none showed recurrence. CONCLUSIONS: Children and adolescents with pathological fractures with features of rickets, recurrent urolithiasis and pancreatitis should be screened for PHPT. Surgery is the treatment of choice. PMID- 19189689 TI - Bone mineral density in girls with classical congenital adrenal hyperplasia due to CYP21 deficiency. AB - AIM: To verify possible associations among glucocorticoid doses, use of dexamethasone, and bone mineral density (BMD), measured by dual X-ray absorptiometry (DXA) and quantitative computed tomography (QCT), in female children with congenital adrenal hyperplasia due to CYP21 deficiency (CAH-CYP21). Classical CAH-CYP21 in females allows the study of the effects of hyperandrogenism and chronic glucocorticoid exposure. DESIGN: Cross-sectional observational study. PATIENTS: Sixteen girls (4-19 years) with CAH-CYP21 and 32 age-matched control girls. MEASUREMENTS: BMD was the main outcome measure assessed by total body and lumbar spine L1-L4 DXA (DXAtot and DXAIs), lumbar spine L1-L4 bone mineral apparent density (BMAD) and spinal L1-L4 QCT of trabecular (QCTtrab) and cortical (QCTcort) bone. The glucocorticoid dose used by patients with CAH-CYP21 was expressed as hydrocortisone equivalents/m2. RESULTS: Mean BMD in both groups was similar by any method. In patients, BMD decreased with the increasing mean dose of glucocorticoid, seen in QCTcort (r = -0.55; p = 0.03) and QCTtrab (r = -0.52; p = 0.04). There was also a negative correlation between cumulative glucocorticoid dose and BMD in QCTcort (beta = -0.0016; p = 0.005) and QCTtrab (beta = -0.0009; p = 0.03). CONCLUSIONS: The dose of glucocorticoid used in the treatment of girls with CAH-CYP21 correlated negatively with BMD, and dexamethasone was not selectively harmful. PMID- 19189690 TI - Economic family burden of metabolic control in children and adolescents with type 1 diabetes mellitus. AB - AIM: To appraise the economic burden for families of patients with type 1 diabetes mellitus (DM1) at the Instituto Nacional de Pediatria in Mexico City. PATIENTS AND METHODS: DM1 family direct costs were obtained from a standardized economic survey in 59 children with no chronic severe complications such as kidney failure in 2002-3. RESULTS: Mean family annual direct cost of treatment and monitoring was US $1689.87 which includes government funding given to both outpatients and inpatients. Despite this, DM1 out-of-pocket cost for families is very high compared to the minimum official wage of approximately $4.00 dollar/day versus $4.06 cost DM1/day. No correlation between parents' age, education or socio-economic level and direct cost was statistically significant. CONCLUSIONS: The highest economic burden was due to self-monitoring of blood glucose (SMBG) 53.0% and insulin 14.8%. The costs were higher in children with poor metabolic control who performed SMBG less often. PMID- 19189691 TI - Simultaneous suprasellar and pineal germ cell tumors in five late stage adolescents: endocrinological studies and prolonged follow-up. AB - Primary germ cell tumors (PGCT) of the central nervous system usually develop in the third ventricle area, and most frequently in the pineal region. The suprasellar region is the second preferential site for development of these tumors which are rarely simultaneously present in these two sites. We report five new cases of PGCT with pineal and suprasellar localizations, which appeared in late puberty in four boys and one girl aged 17-19 years. The clinical picture associated signs of intracranial hypertension, convergence and verticality palsies, diabetes insipidus and pituitary deficiency. Encephalic MRI revealed a double localization. Endocrine tests revealed a particular pattern associating central diabetes insipidus and a hypothalamic-pituitary disconnection syndrome. Following identification of the pathological type of lesions via a neurosurgical approach, treatment was based on a combined method using chemotherapy, radiotherapy and hormone replacement. Based on this treatment, prolonged remissions were obtained with a good quality of life. PMID- 19189692 TI - Thyroidectomy in a patient with multinodular dyshormonogenetic goitre--a case of Pendred syndrome confirmed by mutations in the PDS/SLC26A4 gene. AB - We report a young woman with genetically confirmed Pendred syndrome and discuss the current therapeutic strategies of dyshormonogenetic goitre. A small diffuse thyroid enlargement developed during infancy and although substitution therapy with L-thyroxine was adequate, it progressed and underwent multinodular transformation. Cervical ultrasound at the age of 22 years demonstrated three solid nodules and fine-needle aspiration biopsy showed a finding typical of follicular adenoma. It is known that dyshormonogenetic goitres have a tendency to grow despite appropriate treatment with L-thyroxine. Management of a patient with Pendred syndrome requires careful follow-up and regular imaging of the thyroid. Although the therapeutic approach to dyshormonogenetic goitres is still controversial, in our patient we chose total thyroidectomy as the most advantageous method to prevent the development of malignancies that may arise more frequently from dyshormonogenetic goitres than from goitres of other aetiologies. PMID- 19189693 TI - A new mitochondrial mutation in a patient with diabetes mellitus, deafness, hydronephrosis and joint contractures. AB - A 28-year-old man presented with multiple flexion contractures of hands and feet, deafness, diabetes mellitus and obstructive nephropathy because of bilateral ureterovesicular stricture. Other prominent clinical findings were short stature, bilateral proptosis, multiple freckles and sacralisation of LS. In order to investigate the role of mitochondrial mutations in various clinical manifestations observed in this patient, we performed mutation screening of 1.6 kb mtDNA around the tRNA (Leu(UUR)) part of 16SrRNA and the ND1 gene. No mutation was present at position 3243 which is associated with diabetes mellitus and deafness. A new point mutation (A/G) at position 4093 of the ND1 gene was found. In conclusion, we found a novel mitochondrial mutation in a patient with diabetes and deafness. This mutation has not been reported before and is the first mutation described in the ND1 gene which is related to insulin dependent diabetes mellitus and deafness and could be specific to the Iranian population. All other unusual clinical findings in this patient can be attributed to the presence of this new mutation. PMID- 19189694 TI - Recurrent acute pancreatitis in a child with primary hyperparathyroidism. AB - Hypercalcemia is an important etiology to consider in the evaluation of acute pancreatitis. Not only is it a treatable cause, but understanding the basis for this etiology may provide new insight into the common biochemical mechanisms involved in the pathogenesis of pancreatitis. We report a case of an 11-year-old girl with hypercalcemia due to primary hyperparathyroidism who developed recurrent pancreatitis. We review clinical and experimental data that implicate hypercalcemia as the cause and discuss mechanisms for the association. PMID- 19189695 TI - Severe acanthosis nigricans in a 17 year-old female with partial lipodystrophic syndrome. PMID- 19189696 TI - Aortic elasticity alterations in obesity--even in children! PMID- 19189697 TI - Correlation of abdominal fat accumulation and stiffness of the abdominal aorta in obese children. AB - AIM: To evaluate abdominal adipose tissue accumulation in obese children by ultrasound measurement and compare with the elastic properties of the abdominal aorta. CHILDREN AND METHODS: A total of 56 obese children and a control group of 30 non-obese children had an ultrasound examination for measurements of thickness of visceral, preperitoneal, and subcutaneous fat as well as abdominal aorta stiffness parameters. Anthropometric measurements and metabolic risk profile were assessed by physical examination and blood tests. RESULTS: Abdominal aorta stiffness parameters were all significantly higher in the obese children than in the controls. Among the adipose tissue compartments, visceral fat thickness was the strongest correlate of abdominal aorta stiffness parameters. Similarly, the correlation between visceral abdominal adipose tissue and fasting insulin levels, HOMA score, and pulse pressure also approached significance. CONCLUSIONS: This study suggests that abdominal adipose tissue accumulation is closely associated with cardiovascular risk factors in obese children, and among abdominal adipose tissue compartments, visceral fat thickness was strongly correlated with the elastic properties of the abdominal aorta. PMID- 19189698 TI - Everyday physical activity and adiposity in Prader-Willi syndrome. AB - The aim of this study was to assess the impact of Prader-Willi syndrome (PWS) on the level of everyday physical activity and to explore whether the activity level is related to adiposity. Measurements were performed with an accelerometry-based Activity Monitor during two consecutive schooldays in 12 children with PWS (7-16 years of age) and in 12 age- and gender-matched, healthy children. Adiposity was assessed by body mass index standard deviation scores and by percentage body fat (dual energy X-ray absorptiometry). Mean duration of dynamic activities (expressed as percentage of 24 h) was lower in children with PWS than in the comparison group (8.7 [2.5]% and 12.0 [3.1]%, respectively; p = 0.01). Six children with PWS had normal activity levels. Physical activity level was not related to adiposity. The results indicate that, as a group, children with PWS have an inactive lifestyle. However, children with PWS cannot be stereotyped as inactive since half of them had normal activity levels. PMID- 19189699 TI - Reassessment of the optimal growth hormone cut-off level in insulin tolerance testing for growth hormone secretion in patients with childhood-onset growth hormone deficiency during transition to adulthood. AB - CONTEXT: Retesting of patients with childhood-onset growth hormone deficiency (CO GHD) is recommended after completion of growth hormone (GH) treatment. AIM: To identify patients who are at risk of persistent GHD, and to evaluate the most reliable cut-off level for GH secretion using the insulin tolerance test (ITT) in the transition from childhood to adulthood. RESULTS: Ninety patients (22 female) with CO-GHD were retested using the ITT 1.1 +/- 1.1 years after discontinuation of therapy. Fifty-eight of 77 patients (75%) initially diagnosed with idiopathic GHD showed normalization of GH secretion. Thirteen patients were diagnosed with multiple pituitary hormone deficiency (MPHD) and diagnosis was reconfirmed in all of them, except for one patient. IGF-I levels of patients with persistent GHD were significantly lower (112 +/- 74 ng/ml [range 22-283] vs 245 +/- 107 ng/ml [range 31-505], p<0.01). IGF-I levels correlated positively with GH peak during ITT (r = 0.54, p <0.01), but the wide range of IGF-I levels shows that IGF-I alone cannot replace retesting in many cases. The best sensitivity and specificity scores were obtained when a GH cutoff level below 5.0 ng/ml for patients in the transition phase was used. ROC analysis demonstrates that ITT is a reliable test for evaluation of GH secretion in the transition phase (ROC-AUC 0.97). CONCLUSIONS: Patients with CO-GHD should be retested after discontinuation of therapy to identify those who may profit from further replacement therapy. Patients with organic or genetic GHD or MPHD are likely to have persistent GHD, whereas 75% of our patients with idiopathic GHD showed normalization of growth hormone secretion. IGF-I levels alone are not reliable in the diagnosis of adult GHD in many cases. The best sensitivity and specificity scores were found when a GH cut-off level below 5.0 ng/ml was used in patients with GHD in the transition from childhood to adulthood. PMID- 19189700 TI - Short stature in partially corrected X-linked severe combined immunodeficiency- suboptimal response to growth hormone. AB - BACKGROUND: X-linked severe combined immunodeficiency (XSCID) results from defects in the common cytokine receptor gamma chain (gamma c) required for signaling by receptors for interleukin (IL)-2, -4, -7, -9, -15, and -21. Following haploidentical bone marrow transplant without myelo-conditioning for XSCID, most patients achieve partial reconstitution often limited to T lymphocytes. Many partially corrected patients manifest extreme short stature (<5th percentile). Previous reports have implicated gamma c in growth hormone (GH) receptor signaling, thus severe growth failure in XSCID may be related to the underlying gamma c defect. AIM: To evaluate the GH/insulin-like growth factor I (IGF-I) axis in three children with XSCID and partial immune reconstitution with profound growth failure. METHODS: The IGF-I generation test was performed by administering recombinant GH subcutaneously for 5 days, and measuring serum levels for IGF-I before GH injection, and on days 5 and 8. RESULTS: Study of the somatotropic axis revealed profoundly diminished IGF-I production following rGH challenge in all three patients. CONCLUSION: The data indicate that the GH/IGF-I axis in these partially corrected XSCID patients with severe short stature is profoundly impaired, and supports previous studies suggesting that the underlying gamma c defect may contribute to the severe growth failure in XSCID. This supports a role for defective gamma c in the extreme short stature of XSCID, and raises the possibility of recombinant IGF-I treatment to bypass this defect. PMID- 19189701 TI - Bone speed of sound curves of twin and singleton neonates. AB - AIM: To evaluate associations between prenatal risk factors, neonatal characteristics and bone development. METHODS: Tibial speed of sound (SOS) of 317 neonates whose gestational ages ranged between 25 and 41 weeks and birth weight between 580 and 4,350 g was measured using quantitative ultrasound. RESULTS: Bone SOS values correlated significantly with gestational age and birth weight. Small for gestational age (SGA) infants had higher bone SOS values than appropriate for gestational age (AGA) infants. Infants born to multiple pregnancies had lower SOS values than singleton infants so two separate bone SOS curves were made: for singleton infants born AGA (n = 152), and for infants of multiple pregnancies born AGA (n = 104). Infants of preeclamptic mothers had significantly higher SOS values than the infants of mothers without preeclampsia. CONCLUSION: This is the first study reporting bone SOS values of Turkish neonates. Prenatal risk factors, such as preeclampsia, multiple pregnancy, and being SGA, affect the bone development of the fetus. PMID- 19189702 TI - Torsion of malignant ovarian tumors in childhood and adolescence. AB - AIM: To investigate the frequency of torsion of malignant ovarian tumors in children and adolescents. METHODS: We evaluated all patients treated for adnexal torsion in a tertiary care referral pediatric institution during the last 20 years. Presentation, tumor markers and pathology reports were evaluated. We reviewed the literature on torsion of malignant ovarian tumors. RESULTS: Ninety two girls (age 6 months to 19 years), 41 of them premenarchal, were surgically treated for adnexal torsion. Symptoms and signs that led to clinical investigation and subsequent surgery were not specific. Histological findings of torsioned masses showed 69 non-neoplasms and 23 tumors, including five malignant. Origin of the malignant disease included four germ cell tumors and one sex-cord stromal tumor. The morphology index score for malignant tumors was > or = 7 in all five patients. Tumor markers were elevated in 12 patients, including four of the patients with malignant tumors. Complete staging was performed in three adolescents with stage Ia, IIa and IIIa of disease. We found tumor origin for 11 previous reported patients with torsion of malignant ovarian tumor, including seven germ cell and four granulosa cell tumors. CONCLUSION: Torsion of malignant ovarian tumors in pediatric and adolescent patients occurs very rarely, but it is nevertheless possible at any stage of disease. The most common torsioned malignant ovarian tumors were of germ cell origin, in both premenarchal and postmenarchal girls. A torsioned adnexal mass with index > or = 7 needs to be considered as a potential malignant tumor. PMID- 19189703 TI - Impaired Sertoli cell function in males diagnosed with Noonan syndrome. AB - In order to study male gonadal function in Noonan syndrome, clinical and laboratory data, including inhibin B, were gathered in nine pubertal males diagnosed with Noonan syndrome. Bilateral testicular maldescent was observed in four, and unilateral cryptorchidism occurred in two. Puberty was delayed in three patients. Luteinising hormone (LH) levels were normal in all patients in our series, while follicle stimulating hormone (FSH) levels were raised in seven. Inhibin B was low in six males and just above the lower limit of normal in two. Importantly, all three men with normal testicular descent displayed signs of Sertoli cell dysfunction, indicating, in contrast to earlier reports, that bilateral cryptorchidism does not seem to be the main contributing factor to impairment of testicular function in Noonan syndrome. These findings suggest different mechanisms of disturbance in male gonadal function, which is frequently associated with Sertoli dysfunction. PMID- 19189704 TI - Thyroid stimulating immunoglobulin is often negative in children with Graves' disease. AB - BACKGROUND: The diagnosis of Graves' disease (GD) is typically confirmed with a thyroid stimulating immunoglobulin (TSI) titer. While TSI is reported to be positive in >90% of patients with GD, our anecdotal experience suggests that TSI negative GD may be more common in children. AIM: To investigate the incidence of TSI negative GD in our population and to compare patients with TSI negative vs TSI positive GD. METHODS: Charts of children with GD seen in our pediatric endocrinology clinic over the past 5 years were reviewed. RESULTS: Fifty-seven patients with GD were included. TSI was negative in 43% of patients. No significant differences were found in children with TSI negative vs TSI positive GD. CONCLUSIONS: In patients with GD, the finding of a negative TSI titer usually creates diagnostic uncertainty. However, the fact that nearly half of our patients with GD were TSI negative suggests that this is common in children. PMID- 19189705 TI - Congenital hypothyroidism in Young-Simpson syndrome. AB - We describe a patient with the clinical spectrum of Young-Simpson syndrome. This rare genetic disorder is characterized by congenital hypothyroidism, mental retardation and blepharophimosis. Young-Simpson syndrome is, at present, poorly known to endocrinologists and pediatricians, and should be included in the differential diagnosis of congenital hypothyroidism. It is important to underline that the association of congenital hypothyroidism, blepharophimosis and ptosis allows an exact clinical diagnosis, since the majority of other clinical aspects are common to other disorders. PMID- 19189706 TI - Congenital hypothyroidism caused by a novel homozygous mutation in the thyroid peroxidase gene. AB - Congenital primary hypothyroidism occurs in one out of 4,000 births. About 20% of cases are due to defects in thyroid hormonogenesis. We report on a German girl with congenital hypothyroidism due to a mutation in the thyroid peroxidase (TPO) gene who had elevated serum levels of thyroglobulin during periods of hyperthyrotropinemia. METHODS: The TPO gene was sequenced directly from genomic DNA. RESULTS: The patient had a novel homozygous mutation (R314W) in the TPO gene. The unaffected parents were non-consanguineous and both heterozygous carriers of the mutation. Fifty normal individuals did not harbor the mutation ruling out a common polymorphism. CONCLUSION: The identified TPO gene mutation (R314W) is very likely the genetic cause for hypothyroidism in the reported child. R314W has not been described before and codes for a presumably inactive TPO molecule. PMID- 19189707 TI - Reversible benign intracranial hypertension in a child with hyperthyroidism. AB - Endocrine dysfunction is a rare but known cause of benign intracranial hypertensio (BIH) in adults. Here we describe a rare case of BIH in the pediatric age group associated with autoimmune hyperthyroidism. A 12-year-old girl presented with a 3-month history of headaches. Ophthalmic examination revealed bilateral papilledema. The ocular findings were otherwise normal, with no exophthalmos. Cranial and orbital magnetic resonance imaging was unremarkable. Lumbar CSF opening pressure in recumbent and relaxed position was elevated (31 cm water). Thyroid hormones fT3 and fT4 were elevated while TSH was completely suppressed. As TSH receptor stimulating antibodies (TSHR-Ab) were elevated Graves' disease was diagnosed. Thyroid suppressive therapy with carbimazole was initiated and supplemented by propranolol. As hyperthyroidism improved over two weeks the headaches subsided and the papilledema slowly resolved over the next 2 months. This case illustrates that hyperthyroidism should be considered as a cause of BIH in children. PMID- 19189708 TI - Possible relationship between ring X chromosome and neonatal hypoglycemia. PMID- 19189709 TI - Brainstem projection of the vestibular nerve in the guinea pig: an HRP (horseradish peroxidase) and WGA-HRP (wheat germ agglutinin-HRP) study. AB - We explored the course and termination of primary vestibular afferent fibers within the brainstem of the guinea pig by means of anterograde axonal transport of horseradish peroxidase (HRP) and wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP). Primary vestibular afferent fibers distribute within the entire vestibular nuclear complex, with the exception of the dorsal part of the lateral vestibular nucleus. The superior vestibular nucleus is characterized by the concentration of terminals within its central part. Although terminal labeling is weaker within the periphery, no area completely lacks primary input. The lateral vestibular nucleus can be divided into a ventral and a dorsal part; within the ventral part small and giant cells receive primary afferent fibers, whereas no significant terminal labeling occurs in the dorsal part. The medial vestibular nucleus shows the most uniform labeling, although the lateral part of its rostral third has a few more terminals than the medial half. Primary projection to the descending vestibular nucleus is widespread, although in its rostrodorsal part it is less impressive. Of the small cell groups commonly associated with the vestibular nuclear complex, only group y receives abundant primary input. Whereas group z completely lacks labeled fibers as well as terminals, single primary axons can be observed passing groups x and f. However, no terminals can be found within the borders of these two cell groups. Scanty projections can be detected within the prepositus hypoglossi nucleus, as well as within the external cuneate nucleus, the cochlear nucleus, the abducent nucleus, and parts of the reticular formation. PMID- 19189710 TI - Shapes and distributions of the catecholamine-accumulating neurons in the rabbit retina. AB - Rabbit retinas were fixed with mixed aldehydes and examined for the fluorescence of catecholamines. Labeled cell bodies were present in the layer of the amacrine cells. A band of fluorescent processes was present in layer 1 of the inner plexiform layer. Weaker labeling was present in two deeper strata, one near the middle of the inner plexiform layer (presumably layer 3) and one at the junction of layers 4 and 5. Immunohistochemistry showed tyrosine hydroxylase (TH) to be present in the same cells and the same strata of the inner plexiform layer as the endogenous catecholamines. Exposing the retina to exogenous dopamine or norepinephrine resulted in stronger labeling in the middle and deep levels of the inner plexiform layer. At the same time a second population of amacrine cell bodies became visible. Catecholamine fluorescence contained in the amacrine cell bodies was used as a guide to their injection with Lucifer yellow CH. The filled dendritic arbors revealed two main types of cells. The type 1 cells are monostratified at the most distal level of the inner plexiform layer. They have relatively uncomplicated, radially branching dendritic trees. They are the cells densely stained by immunohistochemistry with antibodies against TH. The type 2 cells are tristratified, with minor branching in layer 1 of the inner plexiform layer and major branching in the two deeper sublayers. The descending dendrites follow a complicated course, and it is not uncommon for intermediate dendrites to cross between strata more than once. The relationship of the cells to their dendritic plexuses was further studied in retinas in which the aldehyde-induced fluorescence of catecholamines was photoconverted to a diaminobenzidine product. The type 1 cells were found to dominate the plexus of dendrites in layer 1 of the inner plexiform layer. The catecholaminergic plexuses in the middle and deep levels of the inner plexiform layer are formed by dendrites of the type 2 cells. The position of every type 1 cell was mapped in retinas stained with antibodies directed against TH. In one retina we counted 5,613 type 1 cells, distributed evenly across the retina. In another retina, all of the catecholamine accumulating cells were counted. There were 9,058 with a distribution that peaks in the visual streak. The type 1 cells appear to be the dopaminergic cells previously studied by others and thought to regulate the flow of information from rod bipolar cells to ganglion cells. The low density and wide spread of type 2 cells suggests that they, too, perform a generalized control function, presumably a novel one that dictates their intricate, tristratified shape. PMID- 19189711 TI - Development and lesion induced reorganization of the cortical representation of the rat's body surface as revealed by immunocytochemistry for serotonin. AB - Immunocytochemistry with an antiserum directed against serotonin (5-HT) was used to assess the development of the representation of the body surface in the rat's primary somatosensory cortex (S-I). Within 1 hour of birth (P-O), 5-HT-positive fibers were present in the marginal zone, the cortical plate, and developing layers V and VI. Immunoreactivity in the marginal zone consisted of a thin band of coarse fibers oriented parallel to the pia. Only a small number of isolated fibers were visible in the cortical plate. A denser network of both coarse and fine fibers could be seen in presumptive layers V and VI. By the first hour of P I, 5-HT-positive axons in the deeper cortical plate were organized into a crude representation of the rat's body surface. At this age, aggregates of fibers corresponding to the head, lower jaw, trunk, and forepaw could be clearly distinguished. These regions of dense 5-HT immunoreactivity consisted primarily of fine caliber axons that had invaded the lower part of the cortical plate. Dense aggregates of fine caliber axons were also visible in developing layers V and VI. Coarse 5-HT-positive fibers were visible in all layers, but they did not appear to contribute to the pattern that corresponded to the body surface. By the first hour of P-2, the map of the body surface in S-I was more refined and a row related organization of 5-HT-immunoreactive fibers was visible in the portion of the cortex representing the vibrissa pad. The laminar distributions of coarse and fine caliber serotoninergic axons at this age were essentially the same as on P I. By P-2.5 (60 hours after birth), patches of 5-HT-positive fibers corresponding to individual vibrissa follicles were clearly evident. These consisted of dense aggregates of fine caliber axons that were centered in presumptive layer IV, but which also extended above and below this lamina. Over the next 3 days, the pattern continued to mature. By P-4, dense 5-HT labelling was also visible in the secondary somatosensory cortex (S-II). By the beginning of P-5, clusters of fibers corresponding to more rostral facial hairs and individual digits within the forepaw representation could also be discerned. By P-12, the differential distribution of 5-HT fibers in S-I was no longer visible. Thus, immunocytochemistry for serotonin showed a representation in S-I homeomorphic with the body surface prior to the age at which it can be discerned with other methods thought to reveal thalamocortical axons. Transection of the infraorbital nerve (ION) on the day of birth altered the organization of the vibrissal representation in the contralateral cortex from the earliest age at which it could be detected by 5-HT immunocytochemistry in normal animals. However, the departure from the normal organization was gradual. Row-related organization was clearly visible in the cortices of rats sacrificed on P-3, but not in those of rats that were killed on P-5. These results suggested that the organization of the 5-HT innervation of the cortex may be guided by thalamic afferents and further that some aspects of this guidance persist, albeit temporarily, after ION transection on P-0. The 5-HT immunoreactivity that we observed in the developing somatosensory cortex was not contained in thalamocortical axons. Unilateral electrocautery of the ventrobasal thalamus on P-4 did not reduce the density or alter the pattern of the 5-HT innervation of the cortex in rats that were examined on P-6. PMID- 19189712 TI - Origin of spinal cord axons in the lizard regenerated tail: supernormal projections from local spinal neurons. AB - During tail regeneration most lizards also regenerate the tail spinal cord. The regenerated spinal cord primarily contains neuroepithelium (i.e., the ependymal tube which forms the central canal) and descending axons. The present experiments identify the source of the axons in the regenerated spinal cord. Application of HRP to normal tail spinal cord resulted in labeled cells in the nucleus paraventricularis, the interstitial nucleus of the fasciculus longitudinalis medialis, the nucleus ruber, the medullary reticular formation (including raphe nuclei), as well as in vestibular nuclei. HRP applied to the regenerated spinal cord labeled only 4% of the cells seen in normal animals, and these were confined to rhombencephalic nuclei. The lack of labeling of more rostral nuclei was not due to the death of descending neurons. Application of HRP immediately rostral to the regenerated spinal cord resulted in the labeling of a normal, and in some cases, greater than normal, number of neurons. To quantify the origin of axons in the regenerated spinal cord, electron microscopic montages of the regenerated spinal cord were made and the number of axons counted, before and after various spinal lesions. Only lesions within one spinal segment of the regenerated spinal cord had a significant effect on the number of axons in the regenerated tail spinal cord. This indicated that most of the regenerated axons were of local spinal origin. A significant increase in the number of labeled local spinal neurons was revealed following application of HRP to a regenerated tail spinal cord. These results suggest that while various portions of the lizard central nervous system can grow axons into the regenerating tail spinal cord, the great majority of axons in the regenerate are of local origin and that some of these arise from neurons that do not normally possess descending projections. Finally, to test whether new neurons were participating in the regeneration process, 3H thymidine was injected during the regrowth of the tail. No labeled spinal cord cells were conclusively identified as neurons. Thus, the regenerating lizard tail spinal cord exhibits robust axonal sprouting from neurons near the site of a spinal transection in a manner reminiscent of sprouting in the mammalian CNS. This sprouting can develop into descending spinal projections that extend for significant distances into the regenerated tail spinal cord and provides a unique model for exploring the requirements for successful axon growth in an adult vertebrate CNS. PMID- 19189713 TI - Innervation of a lizard auditory organ having gap junctions between most hair cells: a serial transmission electron microscopy study. AB - Two apical unidirectional and 16 basal bidirectional papillar hair cells of the yucca night lizard, Xantusia vigilis, were serially sectioned for transmission electron microscopy (TEM) to determine the pattern of hair cell innervation. The 16 bidirectional hair cells (central group) were sectioned across the entire width of the papilla and consisted of four complete hair cells in each of the first three rows and the upper (or neural) half of the four hair cells in the fourth or last row. Both hair cell types were nonexclusively innervated, i.e., each afferent nerve fiber innervated two or more hair cells. The apical unidirectional hair cells were innervated by six or seven different afferent nerve fibers and five or six efferent fibers. The afferent nerve fibers made an average of 52.5 synapses/hair cell. In the central group of 16 bidirectional hair cells, 25 different afferent nerve fibers innervated an average of 4.5 hair cells. The average number of hair cells innervated by the eight afferent nerve fibers limited to the central group was 5.4. An unusual finding was the presence of gap junctions directly interconnecting more than half the hair cells in both papillar segments. In the bidirectional hair cell region, it was possible to count the number of gap junctions between 24 contiguous hair cells. The average number of gap junctions was four per hair cell, and all bidirectional hair cells were either directly or indirectly interconnected by gap junctions. The possible functions of a nonexclusive type of hair cell innervation and the presence of large numbers of gap junctions are discussed. PMID- 19189714 TI - Development of olivocerebellar fibers in the clawed toad, Xenopus laevis: a light and electron microscopical HRP study. AB - An anterograde tracer study was undertaken to provide a light- and electron microscopical description of climbing fiber development in the clawed toad, Xenopus laevis, ranging from premetamorphic stages to the adult state. The inferior olive was unilaterally labeled with horseradish peroxidase and the contralateral climbing fiber morphology investigated. At early stages of development, only undifferentiated fibers were observed in the rostral alar plate. At later stages, these fibers form large varicosities, which contact presumed cerebellar Purkinje cells. Finger-like protrusions arising from the Purkinje cell somata penetrate the climbing fiber varicosities and form synaptic specializations at these contact sites. In older tadpoles, a large variety of climbing fiber morphologies was found displaying a mediolateral gradient. At dorsolateral cerebellar areas long and straight climbing fibers follow the Purkinje cell primary dendrites. However, in ventromedial areas pericellular baskets or nests were found on presumed Purkinje cell somata. These pericellular nests were found throughout development but were not observed in adult animals. Both pericellular nests and real climbing fibers make synaptic contacts on spiny protrusions of the Purkinje cell's somatic or dendritic surface. In several cases, labeled as well as unlabeled climbing fiber profiles were observed on the same Purkinje cell, indicating multiple, convergent innervation. Also, divergent Purkinje cell innervation was found. In conclusion, this study shows that anuran climbing fiber development encompasses stages and processes similar to those observed in mammals. The only principal difference with climbing fiber development in mammals is the low degree of synchrony observed in anurans. PMID- 19189715 TI - Morphological variability is a characteristic feature of granule cells in the primate fascia dentata: a combined Golgi/electron microscope study. AB - This study analyzes the structural variability of granule cells in the monkey fascia dentata. The hippocampi of three adult rhesus monkeys (Macaca mulatta) and two 1-year-old female baboons (Papio anubis) were used for a combined Golgi/electron microscope (EM) study. The results were compared with other Golgi/EM studies on dentate granule cells in small laboratory animals. Whereas the granule cells in small rodents form a relatively uniform population of neurons, we observed a much greater variability of granule cell morphology in monkeys. This variability concerned the size of the cell body, the length and thickness of apical dendrites, the spine density, and the occasional occurrence of basal dendrites. The dendritic length of individual granule cells largely depended on their position in the highly convoluted granular layer. These convolutions caused significant variations in the thickness of the molecular layer and consequently in the length of individual granule cell dendrites. Granule cells with thick dendrites densely covered with spines could be differentiated from those exhibiting much thinner dendritic processes and moderate spine numbers. About 10% of granule cells in the monkey fascia dentata exhibited basal dendrites. Occasionally in the hilus ectopic granule cells were observed that gave rise to long apical dendrites traversing the granular layer. The axons of granule cells, the mossy fibers, entered the hilus, where they gave off several collaterals. In contrast to the light microscopic variability, subtypes of granule cells revealed similar fine structural characteristics, i.e., a round nucleus lacking indentations, a thin rim of cytoplasm, and characteristic spine formations. Large complex spines and smaller, "stubby" spines were observed on apical as well as basal dendrites. This suggests that characteristic spine formations were not induced by specific afferent fibers. All synaptic contacts on spines were of the asymmetric type, whereas both symmetric and asymmetric synapses occurred on cell bodies and dendritic shafts. Unlike in rodents, we found a large variability of granule cells in the primate fascia dentata. This variability has to be considered in neuropathological studies of this cell type. PMID- 19189716 TI - Morphology and cellular interactions of growth cones in the developing corpus callosum. AB - Previous studies of growth cones in invertebrates have shown that they become larger and more complex when changing direction in response to cell-specific contacts (Bentley and Caudy, '83; Raper et al., '83b; Caudy and Bentley, '86). In pathways of the vertebrate nervous system, analogous regions, termed "decision regions," have been identified in which axons change direction and their growth cones become more elaborate than when tracking along straight trajectories (Tosney and Landmesser, '85a; Bovolenta and Mason, '87). In order to assess the generality of these principles to the mammalian CNS, we studied the morphology of growth cones and their interactions with the environment in the developing corpus callosum. Given the straight pathway that callosal axons could use to navigate across the callosum, one might predict that later arriving axons would extend on those growing out earlier and that therefore, by analogy with previous studies, many growth cones would have simple tapered morphologies. Surprisingly, however, virtually all growth cones in the callosal white matter, regardless of age or position, were complex with broad lamellipodial veils and/or numerous, often lengthy filopodia. Only growth cones entering the cortical target were consistently smaller. As seen in the EM, the predominant elements in the callosal pathway are other axons and growth cones; we found no evidence for specialized contacts. These results suggest that there is no specific decision region for the fiber population as a whole; rather it is possible that in this mammalian CNS pathway individual growth cones respond independently to molecular cues broadly distributed in the callosum. PMID- 19189717 TI - Topographic organization of the ventral striatal efferent projections in the rhesus monkey: an anterograde tracing study. AB - The ventral striatum is considered to be that portion of the striatum associated with the limbic system by virtue of its afferent connections from allocortical and mesolimbic areas as well as from the amygdala. The efferent projections from this striatal region in the primate were traced by using 3H amino acids and Phaseolus vulgaris-leucoagglutinin (PHA-L). Particular attention was paid to the topographic organization of terminal fields in the globus pallidus and substantia nigra, the projections to non-extrapyramidal areas, the relationship between projections from the nucleus accumbens and the other parts of the ventral striatum, and the comparison between ventral and dorsal striatal projections. This study demonstrates that in monkeys a circumscribed region of the globus pallidus receives topographically organized efferent fibers from the ventral striatum. The ventral striatal fibers terminate in the ventral pallidum, the subcommissural part of the globus pallidus, the rostral pole of the external segment, and the rostromedial portion of the internal segment. The more central and caudal portions of the globus pallidus do not receive this input. This striatal output appears to remain segregated from the dorsal striatal efferent projections to pallidal structures. Fibers from the ventral striatum projecting to the substantia nigra are not as confined to a specific region as those projecting to the globus pallidus. Although the densest terminal fields occur in the medial portion, numerous fibers also extend laterally to innervate the dorsal stratum of dopaminergic neurons of the substantia nigra and the retrorubral area. Furthermore, they project throughout the rostral-caudal extent of the substantia nigra. Projections from the medial part of the ventral striatum reach the more caudally located pedunculopontine tegmental nucleus. Thus unlike the above described terminals in the globus pallidus, the ventral striatum project widely throughout the substantia nigra, a fact that indicates that they may contribute to the integration between limbic and other output systems of the striatum. Finally, the ventral striatum projects to non-extrapyramidal regions including the bed nucleus of the stria terminals, the nucleus basalis magnocellularis, the lateral hypothalamus, and the medial thalamus. PMID- 19189718 TI - Supplementary eye field as defined by intracortical microstimulation: connections in macaques. AB - In macaques, the frontal eye field and the recently defined supplementary eye field play a role in the production of eye movements. Whereas the structure and function of the frontal eye field are well understood, little is known about the supplementary eye field. The goal of this study was to determine the connections of the physiologically defined supplementary eye field. In each case, the location of the supplementary eye field was determined by using intracortical microstimulation, the borders were marked with small electrolytic lesions, and horseradish peroxidase conjugated to wheat germ agglutinin was injected into the supplementary eye field. After the tissue was incubated with tetramethyl benzidine, it was determined that in three cases the injection site was confined to the physiologically defined supplementary eye field. The present results indicate that the supplementary eye field is reciprocally connected with the claustrum, ventral anterior nucleus, including pars magnocellularis, nucleus X, posterior subdivision of the ventral lateral nucleus, multiform, parvocellular, magnocellular, and densocellular subdivisions of the medial dorsal nucleus, central lateral nucleus, parafascicular nucleus, and suprageniculate-limitans complex. The supplementary eye field projects to the putamen, caudate, reticular nucleus of the thalamus, central densocellular nucleus, zona incerta, subthalamic nucleus, rostral interstitial nucleus of the medial longitudinal fasciculus, parvocellular part of the red nucleus, intermediate and deep layers of the superior colliculus, central gray, cuneiform nucleus, mesencephalic reticular formation, pontine gray, nucleus reticularis tegmenti pontis, and nucleus reticularis pontis oralis. The supplementary eye field is reciprocally and bilaterally connected with periprincipal and inferior prefrontal cortex, with periarcuate cortex, including the frontal eye field, the frontal ventral region, and with postarcuate premotor cortex, and cortex surrounding the supplementary eye field, including the supplementary motor area. The supplementary eye field is also reciprocally connected ipsilaterally with cortex in and around the cingulate sulcus and the intraparietal sulcus, whereas cortex within the superior temporal sulcus projects to the supplementary eye field. The connections of the physiologically defined supplementary eye field are compared to previously demonstrated connections of the supplementary motor region and of the physiologically defined frontal eye field. Comparisons between the connections of the frontal and supplementary eye fields reveal that both regions are connected with structures related to visuomotor functions, but the frontal eye field has more extensive connections with vision-related structures, and the supplementary eye field has more extensive connections with structures related to prefrontal and skeletomotor functions. Such connectional differences suggest functional differences between these two sensorimotor regions of the frontal lobe. PMID- 19189719 TI - Vital queries in medical anthropology: still goaded by the "person". PMID- 19189720 TI - On recognition, caring, and dementia. AB - The onset of dementia raises troubling questions. Does the person with dementia still recognize you? If someone cannot recognize you, can they still care about you? This essay takes such questions as the entry point for a broader inquiry into recognition, its linkages to care, and how claims to social and political "recognition" are linked to, or premised on, the demonstrated capacity to "recognize" people and things. In the words and actions of her severely impaired mother, the author finds guidance toward a better, more compassionate question to ask about dementia: how can we best strive to "keep the cares together"? PMID- 19189721 TI - Politics of care: commentary on Janelle S. Taylor, "On recognition, caring, and dementia". PMID- 19189722 TI - Paternity for sale: anxieties over "demographic theft" and undocumented migrant reproduction in Germany. AB - Women's experiences of migration, and their relationship to a host country, vary significantly from those of migrant men simply because pregnancy is a possibility. The concept of "demographic theft" highlights popular anxieties regarding high fertility among foreigners, including undocumented migrants. This article examines pregnant undocumented women's experiences with the health care system and relationship to the state in Germany. It also provides a discussion of how a restrictive immigration climate, particular models of citizenship, and liberal family laws have resulted in unique practices surrounding paternity claims. It is based on long-term ethnographic data to highlight contradictions and ambiguities in the policy environment and utilizes the notion of stratified reproduction to bring new evidence regarding mothers' deportability and practices of paternity. PMID- 19189723 TI - Cancer rehabilitation in Denmark: the growth of a new narrative. AB - A fundamental assumption behind cancer rehabilitation in many Western societies is that cancer survivors can return to normal life by learning to deal with the consequences of their illness and their treatment. This assumption is supported by increasing political attention to cancer rehabilitation and a growth in residential cancer-rehabilitation initiatives in Denmark (Danish Cancer Society 1999; Government of Denmark 2003). On the basis of their ethnographic fieldwork in residential-cancer rehabilitation courses, the authors examine the new rehabilitation discourse. They argue that this discourse has challenged the dominant illness narrative, "sick-helped-cured," producing a new narrative, "sick helped-as if cured," and that this new narrative is produced and reproduced through technologies of power and of the self. PMID- 19189724 TI - Stigma despite recovery: strategies for living in the aftermath of psychosis. AB - In this article, we identify an array of creative strategies used by persons diagnosed with schizophrenia-related illness to deflect and resist social stigma, and address the lived experience of deploying these strategies in the intersubjective context of everyday life. The data are derived from anthropological interviews and ethnographic observations of ninety persons who received treatment at community mental health facilities in an urban North American locale. Nearly all were keenly aware of stigma that permeated their lives. Their predicament is contradictory: on the one hand, they have recovered relative to previous states of psychosis; on the other hand, their subjectivity is saturated by intense awareness of social stigma that seems intractable in relation to temporal or functional criteria. Ironically, these lives can be characterized as fraught with stigma despite recovery. The strategies generated to resist the impact of stigma highlight the fact that persons with these illnesses are often not only exceedingly socially aware but also strategically skilled in response to social assaults on their personhood and survival. We examine these strategies in terms of (1) the social characteristics of each afflicted person, (2) the situational characteristics of managing stigma, (3) the cultural context of recovery, and (4) the illness-specific characteristics of schizophrenia. PMID- 19189725 TI - What can critical medical anthropology contribute to global health? A health systems perspective. PMID- 19189726 TI - Medical anthropology against war. PMID- 19189727 TI - Visual purple (sehpurpur). PMID- 19189728 TI - Multiplicative and additive Adelson's snake illusions. AB - Two different versions of Adelson's snake lightness illusion are quantitatively investigated. In one experiment an additive version of the illusion is investigated by varying the additive component of the atmosphere transfer function (ATF) introduced by Adelson [2000, in The New Cognitive Neuroscience Ed. M Gazzaniga (Cambridge, MA: MIT Press) pp 339-351]. In the other, a multiplicative version of the illusion is examined by varying the multiplicative component of the ATE In both experiments four observers matched the targets' lightness of the snake patterns with Munsell samples. Increasing the additive or the multiplicative component elicited an approximately equal increase in the magnitude of the lightness illusion. The results show that both components, in the absence of other kinds of information, can be used as heuristics by our visual system to anchor luminance of the object when converting it into lightness. PMID- 19189729 TI - Smiling reduces masculinity: principal component analysis applied to facial images. AB - We examined the effect of facial expression on the assignment of gender to facial images. A computational analysis of the facial images was applied to examine whether physical aspects of the face itself induced this effect. Thirty-six observers rated the degree of masculinity of the faces of 48 men, and the degree of femininity of the faces of 48 women. Half of the faces had a neutral facial expression, and the other half was smiling. Smiling significantly reduced the perceived masculinity of men's faces, especially for male observers, whereas no effect of smiling on femininity ratings was obtained for women's faces. A principal component analysis was conducted on the matrix of pixel luminance values for each facial image x all the images. The third principle component explained a relatively high proportion of the variance of both facial expressions and gender of face. These results suggest that the effect of smiling on the assignment of gender is caused, at least in part, by the physical relationship between facial expression and face gender. PMID- 19189730 TI - Change of temporal-order judgment of sounds during long-lasting exposure to large field visual motion. AB - The perceived temporal order of external successive events does not always follow their physical temporal order. We examined the contribution of self-motion mechanisms in the perception of temporal order in the auditory modality. We measured perceptual biases in the judgment of the temporal order of two short sounds presented successively, while participants experienced visually induced self-motion (yaw-axis circular vection) elicited by viewing long-lasting large field visual motion. In experiment 1, a pair of white-noise patterns was presented to participants at various stimulus-onset asynchronies through headphones, while they experienced visually induced self-motion. Perceived temporal order of auditory events was modulated by the direction of the visual motion (or self-motion). Specifically, the sound presented to the ear in the direction opposite to the visual motion (ie heading direction) was perceived prior to the sound presented to the ear in the same direction. Experiments 2A and 2B were designed to reduce the contributions of decisional and/or response processes. In experiment 2A, the directional cueing of the background (left or right) and the response dimension (high pitch or low pitch) were not spatially associated. In experiment 2B, participants were additionally asked to report which of the two sounds was perceived 'second'. Almost the same results as in experiment 1 were observed, suggesting that the change in temporal order of auditory events during large-field visual motion reflects a change in perceptual processing. Experiment 3 showed that the biases in the temporal-order judgments of auditory events were caused by concurrent actual self-motion with a rotatory chair. In experiment 4, using a small display, we showed that 'pure' long exposure to visual motion without the sensation of self-motion was not responsible for this phenomenon. These results are consistent with previous studies reporting a change in the perceived temporal order of visual or tactile events depending on the direction of self-motion. Hence, large-field induced (ie optic flow) self-motion can affect the temporal order of successive external events across various modalities. PMID- 19189731 TI - The effect of horizontal versus vertical task presentation on children's performance in coordinate tasks. AB - Empirical work on children's ability to understand spatial coordinates has focused on the factors that increase children's proficiency. When interpreting performance, it should be considered that presenting a coordinate task on a horizontal surface might constrain the responses that children make because some target positions are further away from the child than others. Vertical task presentation removes this constraint. Children aged 3 to 9 years were presented with an interpretative coordinate task administered on a touchscreen, presented in an egocentric-vertical position or egocentric-horizontal position. The results show that for 5- to 7-year-old children vertical presentation led to far more correct responses than horizontal presentation. Analysis of the children's errors suggests that this may be due to the fact that vertical presentation suppresses children's bias towards responding in relation to one rather than both coordinates. Taken together these findings contribute to understanding why children's performance in xy coordination tasks is highly contextually sensitive. PMID- 19189733 TI - Mechanisms of identity and gender decisions to faces: who rocked in 1986? AB - Two experiments are reported in which participants made familiarity decisions (Is this face familiar or not?) or gender decisions (Is this face male or female?) to the same sets of faces presented as whole faces or as internal features only. The experimental items on which the analysis was performed were famous and unfamiliar male faces that differed on rated masculinity. The famous faces also differed on age of acquisition (AoA). The experimental male faces were combined with an equal number of famous and unfamiliar female faces for presentation to participants. In experiment 1 we found faster and more accurate familiarity decisions to whole male faces than to internal features; also an interaction between AoA and masculinity such that familiarity decisions were both faster and more accurate to high- than to low-masculinity faces when those faces were late-acquired, but not when they were early-acquired. In experiment 2 we found the same benefit for whole faces over internal features and the same interaction between AoA and masculinity in gender decisions. The similarity between the effects of AoA and masculinity in familiarity and gender decisions is more readily accounted for by models of face processing which posit a close relationship between gender and identity processing than by models which maintain that those aspects of face perception are dealt with by quite separate processing streams. We propose that gender decisions, like familiarity decisions, are semantic judgments (rather than judgments based on the analysis of the surface features of faces), and that the shared basis of the two forms of decision explains why they show similar influences of AoA and masculinity. PMID- 19189732 TI - Learning building layouts with non-geometric visual information: the effects of visual impairment and age. AB - Previous studies suggest that humans rely on geometric visual information (hallway structure) rather than non-geometric visual information (eg doors, signs, and lighting) for acquiring cognitive maps of novel indoor layouts. In this study we asked whether visual impairment and age affect reliance on non geometric visual information for layout learning. We tested three groups of participants-younger (<50 years of age) normally sighted; older (50-70 years of age) normally sighted; and low-vision (people with heterogeneous forms of visual impairment ranging in age from 18 to 67 years). Participants learned target locations in building layouts using four presentation modes: a desktop virtual environment (VE) displaying only geometric cues (sparse VE); a VE displaying both geometric and non-geometric cues (photorealistic VE); a map; and a real building. Layout knowledge was assessed by map drawing and by asking participants to walk to specified targets in the real space. Results indicate that low-vision and older normally sighted participants relied on additional non-geometric information to accurately learn layouts. In conclusion, visual impairment and age may result in reduced perceptual and/or memory processing that makes it difficult to learn layouts without non-geometric visual information. PMID- 19189734 TI - Metaphoric pictures devised by an early-blind adult on her own initiative. AB - EW, a totally blind adult, was encouraged to draw. Three years later, she took a sketchpad on a vacation trip to Mexico and made a series of drawings. In these pictures, objects, such as a glass or a person swimming, are drawn in a realistic manner in which lines stand for surface edges, and the shapes copy parts of the true forms of the objects. Of major interest, EW made drawings in which the lines and forms stood for the effect of the alcoholic liquid in the glass, the taste of a hot pepper, the sound of a trumpet, and the feeling of water running through the fingers of the swimmer. It is suggested that EW has invented apt metaphoric devices on her own initiative. PMID- 19189735 TI - Training eye movements: can training people where to look hinder the processing of fixated objects? AB - An experiment designed to test the effects of different forms of training in a visual-search-like task is reported. Observers were presented with a series of displays containing a central letter and a ring of peripheral characters, one of which was a digit. Odd digits (catch trials) required a space-bar response; even digits required a different response contingent on the identity of the central letter. Two forms of training provided information either about the location of the peripheral digit, or about a quick way to classify the central letter. The aim was to relate training to Findlay and Walker's (1999, Behavioral and Brain Sciences 22 661-721) model of saccadic eye-movement control by affecting the hypothesised Move and Fixate centres respectively. The results showed that training benefited search, but training of the Move centre alone generated significantly longer re-inspections of the central region (in a feedback condition). This highlights that the emphasis often placed upon broadening the range of visual search when training eye movements may be misplaced. More specifically, special attention should be given, not only to advising people how to move their eyes, but also to improving the ability to effectively process important visual stimuli when fixating. PMID- 19189736 TI - Wide and diffuse perceptual modes characterize dyslexics in vision and audition. AB - We examined the performance of dyslexic and typically reading children on two analogous recognition tasks: one visual and the other auditory. Both tasks required recognition of centrally and peripherally presented stimuli. Dyslexics recognized letters visually farther in the periphery and more diffuse near the center than typical readers did. Both groups performed comparably in recognizing centrally spoken stimuli presented without peripheral interference, but in the presence of a surrounding speech mask (the 'cocktail-party effect') dyslexics recognized the central stimuli significantly less well than typical readers. However, dyslexics had a higher ratio of the number of words recognized from the surrounding speech mask, relative to the ones from the center, than typical readers did. We suggest that the evidence of wide visual and auditory perceptual modes in dyslexics indicates wider multi-dimensional neural tuning of sensory processing interacting with wider spatial attention. PMID- 19189737 TI - Blurry means good focus: myopia and visual attention. AB - A correlation between myopia and visuo-spatial attention is reported. More severe myopia was found to be associated with better ability to quickly narrow the focus of visual attention to a small region of space (assessed via interference from spatial proximity of to-be-ignored inverted half-faces), in a task where local focus was explicitly required. There was no myopia association with size of the default attentional window, when the need to respond to either small local or larger global regions was equally likely (in a particular Navon figure task). Results suggest that myopics might allocate attention more narrowly than individuals with normal eyesight in certain functionally important visual tasks (eg reading) but not others (eg driving). PMID- 19189738 TI - The Leaning Tower illusion is not a simple perspective illusion. AB - The Learning Tower illusion has been explained as a simple perspective illusion. I suggest that it is a variant of the Jastrow illusion, applied to perspective tilt, and that the original explanation is inconsistent with its own implicit assumptions and with the visual resolution of pictorial stimuli in general. PMID- 19189739 TI - Using our political weight. PMID- 19189740 TI - Quiz. A 15-year-old boy attends for a routine dental examination. PMID- 19189741 TI - FAQs on Dental Council Code of Conduct. PMID- 19189742 TI - Tooth sensitivity increasing from brushing too hard. PMID- 19189743 TI - The influencer. PMID- 19189744 TI - Dental care with a smile: the CEKA Dental Volunteer Programme in Kenya. PMID- 19189745 TI - Commission issues and infective endocarditis survey. PMID- 19189746 TI - Infective endocarditis prophylaxis and the current AHA, BSAC, NICE and Australian guidelines. AB - The latest guidelines from the American Heart Association (AHA) 2007, the Journal of the American Dental Association (JADA) 2008, the Australian Prevention of Endocarditis Guidelines 2008, the British Society for Antimicrobial Chemotherapy (BSAC) 2006, and the National Institute for Clinical Excellence (NICE) 2008 were reviewed for this article. As a result of recent literature reviews by the AHA and NICE committees, both groups made recommendations regarding antibiotic prophylaxis for dental treatment. While both agree that the benefit of prophylaxis for dental treatment is unproven, the NICE committee has recommended no antibiotic cover for any patients previously classified as 'at risk' of infective endocarditis (IE), while the AHA has recommended cover only for patients deemed to be at high risk of developing IE and with the poorest outcome in the event of IE development. The BSAC guidelines and the recently published Australian Therapeutic Guidelines on Prevention of Endocarditis 2008 fall broadly into line with the AHA guidelines. This paper will review all the separate guidelines and advocate a regimen for treating at-risk patients. PMID- 19189747 TI - Dentists in the DTSS between July 2007 and July 2008. AB - Recent attention in the media concerning the Dental Treatment Services Scheme (DTSS) centres on the number of contracting dentists as a surrogate measure of the availability of services to adult medical card holders in the Republic of Ireland. AIM: To determine the trend in the number of contracting dentists on the DTSS panel during the year 07/'07 to 07/'08. METHODS AND DATA: Data were extracted from the database of monthly claims for remuneration, submitted by providers, which is held by the HSE. RESULTS: The average number of contractors was 1,258. The trend over the year was a reduction of 1.6% in contractor numbers. The average number of 'active' contractors was 833. The trend over the year was a reduction in 'active' contractor numbers of 6.2%. In any month, approximately 34% of contracting dentists were not active. CONCLUSIONS: The trend in the number of contracting dentists was a poor indicator of the trend in availability of services to medical card holders. A better approach would be to count the number of 'active' contractors, in conjunction with their geographic spread and medical card holder density. PMID- 19189748 TI - Quality assurance in dental radiography: intra-oral image quality analysis. AB - With the introduction of criteria for clinical audit by the Irish Dental Council, and the statutory requirement on dentists to introduce this into their practice, this article will introduce the basic concepts of quality standards in intra-oral radiography and the subsequent application of these standards in an image quality audit cycle. Subjective image quality analysis is not a new concept, but its application can prove beneficial to both patient and dental practitioner. The ALARA (as low as reasonably achievable) principle is fundamental in radiation protection, and therefore the prevention of repeat exposures demonstrates one facet of this that the dental practitioner can employ within daily practice. PMID- 19189749 TI - Illuminating magnification. AB - The DOM has few disadvantages but there is a steep learning curve involved for the new user. Undertaking a training course on the use of the microscope before using it in the clinical situation is highly recommended. There are cost implications also, but entry-level microscopes are competitively priced. PMID- 19189750 TI - Development of a novel odor measurement system using gas chromatography with surface acoustic wave sensor. AB - This paper describes a novel odor measurement system for creating arrays of virtual chemical sensors with nonoverlapping responses using ultrahigh-speed gas chromatography with a surface acoustic wave sensor (GC/SAW). This GC/SAW system provides high-resolution two-dimensional olfactory images for easy recognition of many complex odors. Separation and quantification of the individual chemicals within an odor is performed in seconds. Using a solid-state mass-sensitive detector, picogram sensitivity, universal nonpolar selectivity, and electronically variable sensitivity are achieved. An integrated vapor preconcentrator coupled with the electronically variable detector allows the system to measure vapor concentrations spanning 6 or more orders of magnitude. The system attributes of high speed, accuracy, and precision provide a cost effective and complimentary tool for traditional sensory evaluations. PMID- 19189751 TI - Estimate of the pollution removed by leaching of standard domestic waste: mass assessments carried out in batches and columns. AB - Batch and column tests allowed estimation of the mobilization of pollution of standard domestic waste in leachate and biogas. Three laboratory tests (biochemical methane potential (BMP), tank leaching test (TLT), and column tests) have been applied to evaluate the emission of pollutants in liquid and/or gas phase from reconstituted municipal solid waste (MSW) on the basis of French waste. In the case of the batch tests (BMP and TLT), BMP tests indicated a maximum organic carbon share produced by waste (biogas potential) equal to 59% of the initial carbon. The maximum quantity of carbon likely to be leached by the waste (TLT) corresponded to 6% of the carbon contained in the waste. On the other hand, during column tests, 3.3% of carbon was leached and 8% of carbon was evacuated in biogas at the end of a 440-day follow-up. It thus appears that the test conditions have a great influence on the remobilization of pollution. In particular it has been proven than the greater the optimization of the liquid/solid ratio, the agitation, the sowing, and the temperature, the greater the pollutant is leached. This study highlights the possible use of batch and column tests to evaluate the pollution risk of a landfill. PMID- 19189752 TI - Air pollution directional risk assessment for siting a landfill. AB - Air pollution directional risk (APDR) is an essential factor to be assessed when selecting an appropriate landfill site. Because air pollutants generated from a landfill are diffused and transported by wind in different directions and speeds, areas surrounding the landfill will be subject to different associated risks, depending on their relative position from the landfill. This study assesses potential APDRs imposed from a candidate landfill site on its adjacent areas on the basis of the pollutant distribution simulated by a dispersion model, wind directions and speeds from meteorological monitoring data, and population density. A pollutant distribution map layer was created using a geographic information system and layered onto a population density map to obtain an APDR map layer. The risk map layer was then used in this study to evaluate the suitability of a candidate site for placing a landfill. The efficacy of the proposed procedure was demonstrated for a siting problem in central Taiwan, Republic of China. PMID- 19189753 TI - Greenhouse gas mitigation-induced rough-interval programming for municipal solid waste management. AB - A greenhouse gas (GHG) mitigation-induced rough-interval programming model is proposed in this study. Components of GHG emission and environmental pollution control are incorporated into the objective function and a series of relevant constraints. To explicitly examine more complexities existing in many parameters, rough intervals are also communicated into the modeling framework. The proposed model presents satisfactory capabilities in analyzing complicated interrelationships among municipal solid waste (MSW) management, climate-change impact, and environmental pollution control. It can also provide optimal allocation schemes and facilitate decision-makers regulating environmentally sustainable strategies. The developed model is then applied to a case study for demonstrating its applicability. Two representative scenarios (relatively representing two potential management policies that may be implemented in the future years) are considered. The results indicate that the developed model presents advantages in mitigating GHG emissions and the associated climate-change impact. The comparison between the GHG mitigation-induced model with and without rough-interval parameters is also investigated. Completely different solutions of the two models imply the significant impact of dual-uncertain information on the system, which can hardly be addressed through the existing optimization approaches. PMID- 19189754 TI - Seasonal variations and evidence for the effectiveness of pollution controls on water-soluble inorganic species in total suspended particulates and fine particulate matter from Xi'an, China. AB - Total suspended particulate (TSP) and particulate matter less than 2.5 microm in aerodynamic diameter (PM2.5) samples were collected over Xi'an for a 1-yr period to characterize the seasonal variations of water-soluble inorganic ions and to evaluate the effectiveness of the pollution policies and controls during the past 10 yr. Mass concentrations of five cations (sodium [Na+], potassium [K+], ammonium [NH4+], calcium [Ca2+], and magnesium [Mg2+]) and four anions (fluoride [F-], chloride [Cl-], nitrate [NO3-], and sulfate [SO4(2-)]) were determined by ion chromatography. The yearly arithmetic-mean mass concentrations of the total measured water-soluble ions in TSP and PM2.5 were 83.9 +/- 58.4 and 45 +/- 34.3 microg x m(-3). The most abundant ions in TSP were SO4(2-), NO3-, Ca2+, and NH4+; whereas in PM2.5 the dominant ions were SO4(2-), NH4 +, and NO3-. Most of the ions were more concentrated in the PM2.5 than in TSP, but two exceptions were Ca2+ and Mg2+. Comparisons of the molar ratios of Mg2+/Ca2+ in TSP indicated that fugitive dust was the main source for these two ions, and the influence of soil dust from outside of the city was most evident during dust storms. The mass concentrations of SO4(2-), NO3-, , NH4+, and K+ in TSP were highest in winter and lowest in spring, but Ca2+ was much higher in spring than other seasons because of suspended mineral dust. In PM2.5, NO3- and K+ also showed winter maxima, but SO4(2-) and NH4+ were highest in summer. Calculations of ion equivalents showed that TSP samples were more alkaline than PM2.5, the latter being weakly acidic in winter and autumn. High sulfur and nitrogen oxidation ratios occurred in summer and autumn, and there was evidence for the formation of ammonium bisulfate in TSP, ammonium sulfate in PM2.5, and ammonium nitrate in both fractions. Comparisons with the results of prior studies indicate that pollution controls in Xi'an have reduced the levels of air pollution over the past 10 yr. The SO4(2-) concentration during the heating season in 2006 was only about one-eighth of that in 1996, and NH4+ decreased to one-ninth of that in 1996. Seasonal variations in the NO3-/SO4(2-) ratio are different than the patterns observed 10 yr ago, suggesting that emission sources have changed, with those from motor vehicles becoming increasingly important. PMID- 19189755 TI - Forecasting daily source air quality using multivariate statistical analysis and radial basis function networks. AB - It is vital to forecast gas and particle matter concentrations and emission rates (GPCER) from livestock production facilities to assess the impact of airborne pollutants on human health, ecological environment, and global warming. Modeling source air quality is a complex process because of abundant nonlinear interactions between GPCER and other factors. The objective of this study was to introduce statistical methods and radial basis function (RBF) neural network to predict daily source air quality in Iowa swine deep-pit finishing buildings. The results show that four variables (outdoor and indoor temperature, animal units, and ventilation rates) were identified as relative important model inputs using statistical methods. It can be further demonstrated that only two factors, the environment factor and the animal factor, were capable of explaining more than 94% of the total variability after performing principal component analysis. The introduction of fewer uncorrelated variables to the neural network would result in the reduction of the model structure complexity, minimize computation cost, and eliminate model overfitting problems. The obtained results of RBF network prediction were in good agreement with the actual measurements, with values of the correlation coefficient between 0.741 and 0.995 and very low values of systemic performance indexes for all the models. The good results indicated the RBF network could be trained to model these highly nonlinear relationships. Thus, the RBF neural network technology combined with multivariate statistical methods is a promising tool for air pollutant emissions modeling. PMID- 19189756 TI - Characteristics of water-soluble ionic species in fine (PM2.5) and coarse particulate matter (PM10-2.5) in Kaohsiung, southern Taiwan. AB - Eleven ionic species and fine and coarse particle mass concentrations of fine (PM2.5) and coarse (PM10-2.5) particulate matter were investigated in Kaohsiung, southern Taiwan. The PM2.5 and PM10-2.5 particulate concentrations were 49-64 and 34-37 microg x m(-3), respectively. Fifty-five to 64% of the particulate matter less than 10 microm in aerodynamic diameter (PM10) mass was attributed to the PM2.5. PM2.5 concentrations at Daliao (a rural and industrial complex area) were higher than at Tzuoying (an urban and industrial complex area). Ionic species contributed 45-53% and 42-45% of PM2.5 and PM10, respectively. Potassium ions (K+), sulfate (SO4(2-)), and ammonium (NH4+) were predominant in PM2.5, whereas sodium, calcium, and magnesium ions were foremost in PM10-2.5. Nitrate (NO3-) existed in both the PM2.5 and PM10-2.5. Chloride (Cl-), NO3-, and NH4+ concentrations were higher at night than during the day, and they were easily transferred into the gas phase by photochemical reactions and temperature-induced volatilization. The NH4+/SO4(2-) ratios were 2.6 and 2.5 at Daliao and Tzuoying, respectively, which indicated that both sampling sites were rich in NH4+. Therefore, ammonium nitrate would be present in the area. PMID- 19189757 TI - Treatment of propylene glycol monomethyl ether acetate in air streams by a biofilter packed with fern chips. AB - This study aimed to develop a biofilter packed only with fern chips for the removal of airborne propylene glycol monomethyl ether acetate (PGMEA). Fern chips could avoid the shortcomings of traditional media, such as compaction, drying, and breakdown, which lead to the performance failure of the biofilters. In addition, the fern chip medium has the following merits: (1) simplicity in composition; (2) low pressure drop for gas flow (<20 mmH2O x m(-1)); (3) simple in humidification, nutrient addition, pH control, and metabolite removal; (4) economical (US$174-385 x m(-3)), and (5) low weight (wet basis around 290 kg x m( 3)). A two-stage downflow biofilter (2.18 m in height and 0.4 x 0.4 m in cross sectional area) was constructed for the performance test. Both stages were packed with fern chips of 0.30 m in height and 0.40 x 0.40 m in cross-section. Results indicate that with operation conditions of media moisture content controlled in the range of 50-74%, media pH of 6.5-8.3, empty bed retention time (EBRT) of 0.27 0.4 min, influent PGMEA concentrations of 100-750 mg x m(-3), volumetric organic loading of <170 g x m(-3) x hr(-1), and nutrition rates of Urea-nitrogen 66 g x m(-3) x day(-3), potassium dihydrogen phosphate (KH2PO4)-phosphorus 13.3 g x m( 3) x day(-3), and milk powder 1.00 g x m(-3) x day(-1), the fern-chip-packed biofilter could achieve an overall PGMEA removal efficacy of around 94%. Instant milk powder or liquid milk was essential to the good and stable performance of the biofilter for PGMEA removal. PMID- 19189758 TI - Differences between weekday and weekend air pollutant levels in Atlanta; Baltimore; Chicago; Dallas-Fort Worth; Denver; Houston; New York; Phoenix; Washington, DC; and surrounding areas. AB - We evaluated day-of-week differences in mean concentrations of ozone (O3) precursors (nitric oxide [NO], nitrogen oxides [NOx], carbon monoxide [CO], and volatile organic compounds [VOCs]) at monitoring sites in 23 states comprising seven geographic focus areas over the period 1998-2003. Data for VOC measurements were available for six metropolitan areas in five regions. We used Wednesdays to represent weekdays and Sundays to represent weekends; we also analyzed Saturdays. At many sites, NO, NOx, and CO mean concentrations decreased at all individual hours from 6:00 a.m. to 3:00 p.m. on Sundays compared with corresponding Wednesday means. Statistically significant (p < 0.01) weekend decreases in ambient concentrations were observed for 92% of NOx, sites, 89% of CO sites, and 23% of VOC sites. Nine-hour (6:00 a.m. to 3:00 p.m.) mean concentrations of NO, NOx, CO, and VOCs declined by 65, 49, 28, and 19%, respectively, from Wednesdays to Sundays (median site responses). Despite the large reductions in ambient NOx and moderate reductions in ambient CO and VOC concentrations on weekends, ozone and particulate matter (PM) nitrate did not exhibit large changes from weekdays to weekends. The median differences between Wednesday and Sunday mean ozone concentrations at all monitoring sites ranged from 3% higher on Sundays for peak 8-hr concentrations determined from all monitoring days to 3.8% lower on Sundays for peak 1-hr concentrations on extreme-ozone days. Eighty-three percent of the sites did not show statistically significant differences between Wednesday and weekend mean concentrations of peak ozone. Statistically significant weekend ozone decreases occurred at 6% of the sites and significant increases occurred at 11% of the sites. Average PM nitrate concentrations were 2.6% lower on Sundays than on Wednesdays. Statistically significant Sunday PM nitrate decreases occurred at one site and significant increases occurred at seven sites. PMID- 19189759 TI - Conversion of methane to liquid products, hydrogen, and ammonia with environmentally friendly condition-based microgap discharge. AB - The conversion of methane to liquid products, hydrogen (H2), and ammonia (NH3) was investigated experimentally using microgap discharge plasma at an environmentally friendly condition. The experimental results indicated that H2 and NH3 were detected as the main gas products. The highest yield and production rate of H2 was 14.4% (v/v) and 2974.6 micromol/min, respectively, whereas the highest yield and production rate of NH3 was 8000 ppm (v/v) and 165.1 micromol/min, respectively. Particularly, the liquid products were collected on the plate and consisted of pyrrole, 2-methyl-1,4-pentadiene, c-amidopyridine, 2,5 dimethylpyrrole, methylpyrazine, 1-hexyne, 1,4-heptadiene, and polycyclic organic compounds. Some liquid products were the intermediates of drug, flavor, dye, and organic synthesis, as well as edible flavor. The collection efficiency in mass and energy efficiency were 26.3% at once and 22.9 g/kWh, respectively. The whole reaction process was considered to be in line with green chemistry principles. PMID- 19189760 TI - Child abuse: how society and careproviders should respond. PMID- 19189761 TI - Clinical ethics and domestic violence: an introduction. PMID- 19189762 TI - The veil of silence around family violence: is protecting patients' privacy bad for health? PMID- 19189763 TI - State codes on intimate partner violence: victimization reporting requirements for healthcare providers. PMID- 19189764 TI - Deadly sins and cardinal virtues in the clinical management of intimate partner violence. PMID- 19189765 TI - A health-based child protection system: studying a change in paradigm. PMID- 19189766 TI - Social problem or medical condition? A response to Krugman's proposal. PMID- 19189767 TI - Ethical dilemmas in coding domestic violence. PMID- 19189768 TI - The spectrum of religion and science in clinical encounters. PMID- 19189769 TI - Ethical challenges of research on and care for victims of intimate partner violence. PMID- 19189770 TI - A human model that suggests a role for sleep in the cognitive neuropsychology of PTSD and recovery. PMID- 19189771 TI - Craniofacial profile assessment in patients with obstructive sleep apnea. PMID- 19189772 TI - Heart failure, central sleep apnea, CPAP, and arousals: another piece of the puzzle. PMID- 19189773 TI - Discovery of the dreaming sleep stage: a recollection. PMID- 19189774 TI - Re-redefining success in airway surgery for obstructive sleep apnea. PMID- 19189775 TI - Sleep promotes generalization of extinction of conditioned fear. AB - STUDY OBJECTIVE: To examine the effects of sleep on fear conditioning, extinction, extinction recall, and generalization of extinction recall in healthy humans. DESIGN: During the Conditioning phase, a mild, 0.5-sec shock followed conditioned stimuli (CS+s), which consisted of 2 differently colored lamps. A third lamp color was interspersed but never reinforced (CS-). Immediately after Conditioning, one CS+ was extinguished (CS+E) by presentation without shocks (Extinction phase). The other CS+ went unextinguished (CS+U). Twelve hours later, following continuous normal daytime waking (Wake group, N=27) or an equal interval containing a normal night's sleep (Sleep group, N=26), conditioned responses (CRs) to all CSs were measured (Extinction Recall phase). It was hypothesized that the Sleep versus Wake group would show greater extinction recall and/or generalization of extinction recall from the CS+E to the CS+U. SETTING: Academic medical center. SUBJECTS: Paid normal volunteers. MEASUREMENTS AND RESULTS: Square-root transformed skin conductance response (SCR) measured conditioned responding. During Extinction Recall, the Group (Wake or Sleep) x CS+ Type (CS+E or CS+U) interaction was significant (P = 0.04). SCRs to the CS+E did not differ between groups, whereas SCRs to the CS+U were significantly smaller in the Sleep group. Additionally, SCRs were significantly larger to the CS+U than CS+E in the Wake but not the Sleep group. CONCLUSIONS: After sleep, extinction memory generalized from an extinguished conditioned stimulus to a similarly conditioned but unextinguished stimulus. Clinically, adequate sleep may promote generalization of extinction memory from specific stimuli treated during exposure therapy to similar stimuli later encountered in vivo. PMID- 19189777 TI - Craniofacial phenotyping in obstructive sleep apnea--a novel quantitative photographic approach. AB - STUDY OBJECTIVES: To compare the craniofacial morphological phenotype of subjects with and without obstructive sleep apnea (OSA) using a quantitative photographic analysis technique. DESIGN: Case-control study; subgroup matched for body mass index (BMI) and sex. SETTING: Sleep investigation unit in a university teaching hospital. PATIENTS: 114 subjects (93% Caucasian) with OSA (apnea-hypopnea index [AHI] > or = 10/h) and 66 controls (AHI < 10/h). INTERVENTIONS: Standardized frontal-profile craniofacial photographic imaging performed prior to polysomnography. Photographs were analyzed for the computation of linear, angular, area and polyhedral volume measurements representing dimensions and relationships of the various craniofacial regions. MEASUREMENTS AND RESULTS: Photographic craniofacial phenotypic differences were demonstrated between OSA and control subjects, including a range of measurements of the face, mandible, maxilla, eyes, nose, head and neck. After 1-for-1 subgroup matching for BMI and sex (51 subjects in each group), mandibular length 1 (6.21 +/- 0.08 [mean +/- SEM] vs. 6.58 +/- 0.08 cm, P = 0.006), mandibular-nasion angle 1 (35.0 +/- 0.48 vs. 36.7 +/- 0.37 degrees, P = 0.006) and anterior neck space area (10.2 +/- 0.53 vs. 12.2 +/- 0.52 cm2, P = 0.01) remained smaller in the OSA group. Mandibular width-length angle (88.0 +/- 0.75 vs. 85.3 +/- 0.54 degrees, P = 0.005) and face width-midface depth angle (72.3 +/- 0.44 vs. 70.7 +/- 0.39 degrees, P = 0.01) remained larger in the OSA group, whereas mandibular triangular area (39.2 +/- 0.63 vs. 41.7 +/- 0.74 cm2, P = 0.01) was smaller. CONCLUSIONS: Craniofacial phenotypic differences in OSA in Caucasian subjects can be demonstrated using a photographic analysis technique. PMID- 19189776 TI - Effects and side-effects of surgery for snoring and obstructive sleep apnea--a systematic review. AB - STUDY OBJECTIVES: Many patients undergo surgery for snoring and sleep apnea, although the efficacy and safety of such procedures have not been clearly established. Our aim was systematically to review studies of the efficacy and adverse effects of surgery for snoring and obstructive sleep apnea. DESIGN: Systematic review. MEASUREMENTS: PubMed and Cochrane databases were searched in September 2007. Randomized controlled trials of surgery vs. sham surgery or conservative treatment in adults, with daytime sleepiness, quality of life, apnea hypopnea index, and snoring as outcomes were included. Observational studies were also reviewed to assess adverse effects. Evidence of effect required at least two studies of medium and high quality reporting the same result. RESULTS: Four studies of benefits and 45 studies of adverse effects were included. There was no significant effect on daytime sleepiness and quality of life after laser-assisted uvulopalatoplasty and radiofrequency ablation. The apnea-hypopnea index and snoring was reduced in one trial after laser-assisted uvulopalatoplasty but not in another trial. Subjective snoring was reduced in one trial after radiofrequency ablation. No trial investigating the effect of any other surgical modality met the inclusion criteria. Persistent side-effects occurred after uvulopalatopharyngoplasty and uvulopalatoplasty in about half the patients and difficulty in swallowing, globus sensation and voice changes were especially common. CONCLUSIONS: Only a small number of randomized controlled trials with a limited number of patients assessing some surgical modalities for snoring or sleep apnea are available. These studies do not provide any evidence of effect from laser-assisted uvulopalatoplasty or radiofrequency ablation on daytime sleepiness, apnea reduction, quality of life or snoring. We call for research of randomized, controlled trials of surgery other than uvulopalatopharyngoplasty and uvulopalatoplasty, as they are related to a high risk of long-term side-effects, especially difficulty swallowing. PMID- 19189778 TI - Prediction of obstructive sleep apnea with craniofacial photographic analysis. AB - STUDY OBJECTIVES: To develop models based on craniofacial photographic analysis for the prediction of obstructive sleep apnea (OSA). DESIGN: Prospective cohort study. SETTING: Sleep investigation unit in a university teaching hospital. PATIENTS: One hundred eighty subjects (95.6% Caucasian) referred for the initial investigation of OSA were recruited consecutively. INTERVENTIONS: Clinical assessment and frontal-profile craniofacial photographic analyses were performed prior to polysomnography. Prediction models for determining the presence of OSA (apnea-hypopnea index [AHI] > or =10) were developed using logistic regression analysis and classification and regression trees (CART). MEASUREMENTS AND RESULTS: Obstructive sleep apnea was present in 63.3% of subjects. Using logistic regression, a model with 4 photographic measurements (face width, eye width, cervicomental angle, and mandibular length 1) correctly classified 76.1% of subjects with and without OSA (sensitivity 86.0%, specificity 59.1%, area under the receiver operating characteristics curve [AUC] 0.82). Combination of photographic and other clinical data improved the prediction (AUC 0.87), whereas prediction based on clinical assessment alone was lower (AUC 0.78). The optimal CART model provided a similar overall classification accuracy of 76.7%. Based on this model, 59.4% of the subjects were classified as either high or low risk with positive predictive value of 90.9% and negative predictive value of 94.7%, respectively. The remaining 40.6% of subjects have intermediate risk of OSA. CONCLUSIONS: Craniofacial photographic analysis provides detailed anatomical data useful in the prediction of OSA. This method allows OSA risk stratification by craniofacial morphological phenotypes. PMID- 19189779 TI - The economic burden of insomnia: direct and indirect costs for individuals with insomnia syndrome, insomnia symptoms, and good sleepers. AB - BACKGROUND AND PURPOSE: Insomnia is a highly prevalent problem that is associated with increased use of health care services and products, as well as functional impairments. This study estimated from a societal perspective the direct and indirect costs of insomnia. PARTICIPANTS AND METHODS: A randomly selected sample of 948 adults (mean age = 43.7 years old; 60% female) from the province of Quebec, Canada completed questionnaires on sleep, health, use of health-care services and products, accidents, work absences, and reduced productivity. Data were also obtained from the Quebec government administered health insurance board regarding consultations and hospitalizations. Participants were categorized as having insomnia syndrome, insomnia symptoms or as being good sleepers using a standard algorithm. Frequencies of target cost variables were obtained and multiplied by unit costs to generate estimates of total costs for the adult population of the province of Quebec. RESULTS: The total annual cost of insomnia in the province of Quebec was estimated at $6.6 billion (Cdn$). This includes direct costs associated with insomnia-motivated health-care consultations ($191.2 million) and transportation for these consultations ($36.6 million), prescription medications ($16.5 million), over the-counter products ($1.8 million) and alcohol used as a sleep aid ($339.8 million). Annual indirect costs associated with insomnia-related absenteeism were estimated at $970.6 million, with insomnia related productivity losses estimated at $5.0 billion. The average annual per person costs (direct and indirect combined) were $5,010 for individuals with insomnia syndrome, $1431 for individuals presenting with symptoms, and $421 for good sleepers. CONCLUSIONS: This study suggests that the economic burden of insomnia is very high, with the largest proportion of all expenses (76%) attributable to insomnia-related work absences and reduced productivity. As the economic burden of untreated insomnia is much higher than that of treating insomnia, future clinical trials should evaluate the cost-benefits, cost-utility, and cost-effectiveness of insomnia therapies. PMID- 19189780 TI - Insomnia did not predict incident hypertension in older adults in the cardiovascular health study. AB - STUDY OBJECTIVE: We hypothesized that the sleep complaints of insomnia predict incident hypertension, particularly in African Americans. The purpose of this study was to analyze insomnia complaints as predictors of incident hypertension in the Cardiovascular Health Study (CHS), stratifying by gender and allowing for race and sleep variable interaction. DESIGN: This is a prospective cohort study over a 6-year period of follow-up. SETTING: This is a community-based study of participants in Forsyth County, North Carolina; Pittsburgh, Pennsylvania; Sacramento County, California; and Washington County, Maryland. PARTICIPANTS: The study analyzed data from 1419 older individuals (baseline mean age 73.4 +/- 4.4 years) from the Cardiovascular Health Study who were not hypertensive at baseline. INTERVENTIONS: none. MEASUREMENTS: We constructed relative risks of incident hypertension over a 6-year period for insomnia complaints singly and in combination. RESULTS: Difficulty falling asleep, singly or in combination with other sleep complaints, predicted a statistically significant reduction of risk for incident hypertension for non-African American men in 6 years of follow-up. Insomnia complaints did not predict incident hypertension in 6 years of follow-up in women or in African Americans, although there may not have been enough power to show a significant association for African Americans. CONCLUSIONS: Insomnia did not predict hypertension in this older cohort which was free of hypertension at baseline. Difficulty falling asleep was associated with reduced risk of hypertension in non-African American men. PMID- 19189782 TI - Development of NREM sleep instability-continuity (cyclic alternating pattern) in healthy term infants aged 1 to 4 months. AB - STUDY OBJECTIVES: To evaluate non-rapid eye movement (NREM) sleep instability, as measured by the cyclic alternating pattern (CAP), in the first months of life in a group of normal healthy infants, in order to obtain more information on the maturation of arousal mechanisms during NREM sleep and to set normative data of CAP parameters in this age range (from 1 to 4 months of life). DESIGN: Retrospective study. SETTING: Sleep unit of an academic centre. PARTICIPANTS: Twenty-three healthy newborns and infants with a mean conceptional age (gestational age plus postnatal age) of 47.6 + 3.8 weeks, age range 42 to 55 weeks, 10 boys (43.47%), were studied while they slept in the morning between feedings, by means of a 3-hour video-electroencephalographic (EEG)-polygraphic recording. Sleep was visually scored for sleep architecture and CAP in a blinded fashion, using standard criteria. MEASUREMENTS AND RESULTS: We found 3 different sleep EEG patterns in our infants, according to their age, and we subdivided the entire group into 3 subgroups. Group 1-Trace alternant mixed with high-voltage slow activity included 9 subjects (3 boys), with a mean conceptional age of 43.9 +/- 1.3 weeks; Group 2 (high-voltage slow activity and rudimentary spindles) included 6 subjects (4 boys), with a mean conceptional age of 49.4 +/- 3.1 weeks; and Group 3 (slow-wave activity and spindles, scored as NREM sleep) included 8 subjects (3 boys), with a mean conceptional age of 50.4 +/- 2.9 weeks. CAP rate was 6.83 +/- 3.58 in infants belonging to Group 2 and increased to 12.91 +/- 2.21 in Group 3. We found a statistically significant higher A1 index in only Group 3. The relative percentages of the A1, A2, and A3 subtypes showed non significant changes with age. The duration of CAP events and the cortical and subcortical arousal indexes were not statistically different between Groups 2 and 3. CONCLUSIONS: With this study, we provide the first data on CAP analysis in infants from 1 to 4 months of life, and we found that there is a transitory period when trace alternant disappears and CAP events begin to occur. Furthermore, we suggest that the more appropriate time of life when CAP analysis can be first performed is related to the appearance of mature stage 2 NREM with spindles and slow delta waves mixed with theta waves, at approximately 3 months of life. PMID- 19189781 TI - Race and financial strain are independent correlates of sleep in midlife women: the SWAN sleep study. AB - STUDY OBJECTIVES: To examine racial differences in sleep in a large cohort of midlife women and to evaluate whether indices of socioeconomic status (SES) are associated with racial differences in sleep. DESIGN: Cross-sectional study. SETTING: Participants' homes. PARTICIPANTS: Caucasian (n=171), African American (n=138) and Chinese women (n=59). INTERVENTIONS: None. MEASUREMENTS: Sleep quality was assessed with the Pittsburgh Sleep Quality Index. Polysomnographically assessed sleep duration, continuity, architecture, and NREM electroencephalograhic (EEG) power were calculated over multiple nights. Sleep disordered breathing and periodic leg movements were measured on a separate night. Linear regression analysis was used to model the independent and synergistic effects of race and SES on sleep after adjusting for other factors that impact sleep in midlife women. Indices of SES were self-reported educational attainment and financial strain. RESULTS: Sleep was worse in African American women than Caucasian participants as measured by self-report, visual sleep stage scoring, and NREM EEG power. Slow wave sleep differences were also observed between Chinese and Caucasian participants. Racial differences persisted after adjustment for indices of SES. Although educational attainment was unrelated to sleep, financial strain was associated with decreased sleep quality and lower sleep efficiency. Financial strain-by-race interactions were not statistically significant, suggesting that financial strain has additive effects on sleep, independent of race. CONCLUSIONS: Independent relationships between race and financial strain with sleep were observed despite statistical adjustment for other factors that might account for these relationships. Results do not suggest that assessed indices of SES moderate the race-sleep relationship, perhaps due to too few women of low SES in the study. PMID- 19189783 TI - Effect of continuous positive airway pressure on sleep structure in heart failure patients with central sleep apnea. AB - STUDY OBJECTIVES: At termination of obstructive apneas, arousal is a protective mechanism that facilitates restoration of upper airway patency and airflow. Treating obstructive sleep apnea (OSA) by continuous positive airway pressure (CPAP) reduces arousal frequency indicating that such arousals are caused by OSA. In heart failure (HF) patients with central sleep apnea (CSA), however, arousals frequently occur several breaths after apnea termination, and there is uncertainty as to whether arousals from sleep are a consequence of CSA. If so, they should diminish in frequency when CSA is attenuated. We therefore sought to determine whether attenuation of CSA by CPAP reduces arousal frequency. DESIGN: Randomized controlled clinical trial. PATIENTS AND SETTING: We examined data from 205 HF patients with CSA (apnea-hypopnea index [AHI] > or =15, > 50% were central) randomized to CPAP or control who had polysomnograms performed at baseline and 3 months later. MEASUREMENTS AND RESULTS: In the control group, there was no change in AHI or frequency of arousals. In the CPAP-treated group, the AHI decreased significantly (from [mean +/- SD] 38.9 +/- 15.0 to 17.6 +/- 16.3, P < 0.001) but neither the frequency of arousals nor sleep structure changed significantly. CONCLUSION: These data suggest that attenuation of CSA by CPAP does not reduce arousal frequency in HF patients. We conclude that arousals were not mainly a consequence of CSA, and may not have been acting as a defense mechanism to terminate apneas in the same way they do in OSA. PMID- 19189784 TI - Irregular respiration as a marker of wakefulness during titration of CPAP. AB - STUDY OBJECTIVES: Regularity of respiration is characteristic of stable sleep without sleep disordered breathing. Appearance of respiratory irregularity may indicate onset of wakefulness. The present study examines whether one can detect transitions from sleep to wakefulness using only the CPAP flow signal and automate this recognition. DESIGN: Prospective study with blinded analysis SETTING: Sleep disorder center, academic institution. PARTICIPANTS: 74 subjects with obstructive sleep apnealhypopnea syndrome (OSAHS) INTERVENTIONS: n/a. MEASUREMENTS AND RESULTS: 74 CPAP titration polysomnograms in patients with OSAHS were examined. First we visually identified characteristic patterns of ventilatory irregularity on the airflow signal and tested their relation to conventional detection of EEG defined wake or arousal. To automate recognition of sleep-wake transitions we then developed an artificial neural network (ANN) whose inputs were parameters derived exclusively from the airflow signal. This ANN was trained to identify the visually detected ventilatory irregularities. Finally, we prospectively determined the accuracy of the ANN detection of wake or arousal against EEG sleep/wake transitions. A visually identified irregular respiratory pattern (IrREG) was highly predictive of appearance of EEG wakefulness (Positive Predictive Value [PPV] = 0.89 to 0.98 across subjects). Furthermore, we were able to automate identification of this irregularity with an ANN which was highly predictive for wakefulness by EEG (PPV 0.66 to 0.86). CONCLUSIONS: Despite not detecting all wakefulness, the high positive predictive value suggests that analysis of the respiration signal alone may be a useful indicator of CNS state with potential utility in the control of CPAP in OSAHS. The present study demonstrates the feasibility of automating the detection of IrREG. PMID- 19189785 TI - Prevalence and correlates of insomnia and its impact on quality of life in Chinese schizophrenia patients. AB - STUDY OBJECTIVES: This study aimed to determine the prevalence and the sociodemographic and clinical correlates of insomnia in Chinese schizophrenia outpatients and its impact on patients' quality of life (QOL). DESIGN: Two hundred fifty-five clinically stable schizophrenia outpatients were randomly selected in Hong Kong and their counterparts matched according to sex, age, age at onset, and length of illness were recruited in Beijing, China. All subjects at both sites were interviewed by the same investigator using standardized assessment instruments. SETTING: Hong Kong and Beijing, China. PATIENTS OR PARTICIPANTS: Clinically stable schizophrenia outpatients. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: In the combined Beijing-Hong Kong sample the frequency of at least one type of insomnia over the previous 12 months was 36.0%; the rates of difficulty initiating sleep (DIS), difficulty maintaining sleep (DMS), and early morning awakening (EMA) were 21.2%, 23.6%, and 11.9%, respectively. Poor sleep was significantly associated with advanced age, older age at onset, fewer psychiatric admissions, severity of positive symptoms, anxiety, extrapyramidal symptoms (EPS) and depressive symptoms, less frequent use of atypical antipsychotic medications (AP), and more frequent use of benzodiazepines (BZD) and hypnotics. Poor sleepers had significantly poorer QOL in all domains than patients without insomnia. After controlling for the potential confounding effects of sociodemographic and clinical factors, a significant difference remained between the 2 groups with regard to the physical QOL domain. A multiple logistic regression analysis found that advanced age, fewer psychiatric admissions, severity of depressive symptoms and use of hypnotics were significant contributors to poor sleep. CONCLUSION: Insomnia is independently associated with poor QOL. More attention should be paid in clinical practice to the high rate of insomnia in Chinese schizophrenia patients. PMID- 19189787 TI - Obstructive apneas induce early release of mesenchymal stem cells into circulating blood. AB - STUDY OBJECTIVES: To investigate whether noninvasive application of recurrent airway obstructions induces early release of mesenchymal stem cells into the circulating blood in a rat model of obstructive sleep apnea. DESIGN: Prospective controlled animal study. SETTING: University laboratory. PATIENTS OR PARTICIPANTS: Twenty male Sprague-Dawley rats (250-300 g). INTERVENTIONS: A specially designed nasal mask was applied to the anesthetized rats. Ten rats were subjected to a pattern of recurrent obstructive apneas (60 per hour, lasting 15 seconds each) for 5 hours. Ten anesthetized rats were used as controls. MEASUREMENTS AND RESULTS: Mesenchymal stem cells from the blood and bone marrow samples were isolated and cultured to count the total number of colony-forming unit fibroblasts (CFU-F) of adherent cells after 9 days in culture. The number of CFU-F from circulating blood was significantly (P = 0.02) higher in the rats subjected to recurrent obstructive apneas (5.00 +/- 1.16; mean +/- SEM) than in controls (1.70 +/- 0.72). No significant (P = 0.54) differences were observed in CFU-F from bone marrow. CONCLUSIONS: Application of a pattern of airway obstructions similar to those experienced by patients with sleep apnea induced an early mobilization of mesenchymal stem cells into circulating blood. PMID- 19189788 TI - Lower urinary tract symptoms and urinary incontinence. PMID- 19189786 TI - A consensus definition of cataplexy in mouse models of narcolepsy. AB - People with narcolepsy often have episodes of cataplexy, brief periods of muscle weakness triggered by strong emotions. Many researchers are now studying mouse models of narcolepsy, but definitions of cataplexy-like behavior in mice differ across labs. To establish a common language, the International Working Group on Rodent Models of Narcolepsy reviewed the literature on cataplexy in people with narcolepsy and in dog and mouse models of narcolepsy and then developed a consensus definition of murine cataplexy. The group concluded that murine cataplexy is an abrupt episode of nuchal atonia lasting at least 10 seconds. In addition, theta activity dominates the EEG during the episode, and video recordings document immobility. To distinguish a cataplexy episode from REM sleep after a brief awakening, at least 40 seconds of wakefulness must precede the episode. Bouts of cataplexy fitting this definition are common in mice with disrupted orexin/hypocretin signaling, but these events almost never occur in wild type mice. It remains unclear whether murine cataplexy is triggered by strong emotions or whether mice remain conscious during the episodes as in people with narcolepsy. This working definition provides helpful insights into murine cataplexy and should allow objective and accurate comparisons of cataplexy in future studies using mouse models of narcolepsy. PMID- 19189789 TI - Quality of care in government family planning clinic services in Colombo District. AB - OBJECTIVE: To assess the quality of care in government family planning clinic services in Colombo District. DESIGN: Descriptive cross-sectional study. SETTINGS: Government family planning clinics in the Colombo District. STUDY SAMPLE: The study was conducted in 23 government family planning clinics in the Colombo District selected through stratified sampling. 593 women visiting these clinics for family planning services were interviewed and 242 client-provider interactions were observed. MEASUREMENTS: Client exit interviews, observations of service delivery and clinic inventories were used as survey tools to assess the quality in preparedness of the clinics, the service delivery process and client satisfaction, as outcomes of the service. A set of indicators were identified for this purpose. RESULTS: The study showed deficiencies in infrastructure, interpersonal relations, privacy, information to clients, especially on side effects and warning signs, and in the mechanisms to ensure continuity. The majority of clients were satisfied with many aspects of the service such as the family planning method received (94.5%), confidentiality of information shared with the provider (96.1%), competency of provider (97.5%), and physical access to clinics (92.3%). However, many were dissatisfied with the physical conditions of the clinics (> 20%), information received (12.5%), opportunity given to discuss their problems with the service providers (18.8%) and waiting times (26.6%). CONCLUSIONS: Government family planning clinic services need improvement through upgrading of clinic infrastructure, better planning and management of clinic services, regular training of service providers and establishing of a system to monitor service quality. PMID- 19189790 TI - Prediction of total and visceral fat contents using anthropometric measures of adiposity in women. AB - BACKGROUND: Although techniques such as dual energy xray absorptiometry (DXA) and quantitative CT are available to estimate global and regional adiposity, anthropometric measurements are often used to detect adiposity in clinical practice. OBJECTIVES: To assess the association between the anthropometric measurements of obesity with total and regional fat mass determined by DXA. DESIGN: A cross-sectional, descriptive study. Patients and method 106 healthy women volunteers, aged between 30 and 54 years were studied. Anthropometric measurements including body mass index (BMI), waist circumference (WC), hip circumference (HC), height adjusted WC (WC/HT), waist-hip ratio (WHR), and skin fold thickness (SFT) over triceps, infrascapular, and iliac regions were made. All women underwent assessment of total fat mass (TFM) and visceral fat mass (VFM) using a Hologic DXA scan. RESULTS: TFM and VFM showed positive correlations with all the anthropometric measurements examined, the strongest correlation was with BMI (r = 0.89 and 0.77 for TFM and VFM respectively, p < 0.001). Correlations of TFM with WC, HC, and WC/HT were 0.72, 0.87, and 0.65, (p < 0001 for all) respectively. Corresponding figures for VFM were 0.73, 0.74, and 0.68, (p < 0001 for all). WHR showed a poor correlation with TFM (r = 18, p = 0.09) and VFM (r = 0.33, p = 0.002). SFTs measured at three sites showed less strong correlations with TFM and VFM (r = 0.48 to 0.69, p < 0.001). CONCLUSIONS: BMI has the strongest association with total and visceral fat mass among these women. Waist and hip circumferences showed high correlations with total and visceral fat mass, but adjusting waist circumference for height did not improve the correlation. PMID- 19189791 TI - An IVF baby with neonatal lupus erythematosus. PMID- 19189792 TI - Hyperhaemolysis syndrome in haemoglobin E / beta thalassaemia responding to cyclophosphamide therapy. PMID- 19189793 TI - Acute systemic paraquat intoxication: survival without long-term complications. PMID- 19189794 TI - Modified simple percutaneous suprapubic cystostomy. PMID- 19189795 TI - A brain bank for Sri Lanka. PMID- 19189796 TI - Paraneoplastic syndromes. PMID- 19189797 TI - Metabolic and endocrine manifestations of neoplastic disease. PMID- 19189798 TI - Cutaneous manifestations of neoplastic disease. PMID- 19189799 TI - Paraneoplastic neurological syndromes. PMID- 19189800 TI - Anal incontinence in the aging Sri Lankan population: the results of a pilot study. PMID- 19189801 TI - Clinical features of leptospirosis: a prospective descriptive study at the National Hospital of Sri Lanka (NHSL) in 2007. PMID- 19189802 TI - Chromobacterium violaceum infection in a provincial hospital in Sri Lanka. PMID- 19189803 TI - Characteristics of out-of-home caregiving environments provided under child welfare services. AB - A national probability sample of children who have been in child welfare supervised placements for about one year identifies the characteristics (e.g., age, training, education, health, and home) of the foster parents, kinship foster parents, and group home caregivers. Caregiving respondents provided information about their backgrounds. Interviewers also used the HOME-SF to assess the caregiving environments of foster care and kinship care. Comparisons are made to other nationally representative samples, including the U.S. Census and the National Survey of America's Families. Kinship care, foster care, and group care providers are significantly different from each other--and the general population -in age and education. Findings on the numbers of children cared for, understimulating environments, use of punitive punishment, and low educational levels of caregivers generate suggestions for practice with foster families. PMID- 19189804 TI - Engaging families in child welfare services: worker versus client perspectives. AB - Part of a larger mixed-method study of engagement in neighborhood-based child welfare services, the qualitative data this article reports on highlights the extent to which parents and workers differ in their views of engagement, the best ways to foster engagement in services, and the importance each group places on it as a process. Strategies designed to improve engagement are offered, including knowledge that can help workers interact more effectively with families and in so doing improve permanency for children. PMID- 19189805 TI - Accessing substance abuse treatment: issues for parents involved with child welfare services. AB - The complex issues associated with barriers to treatment entry for parents who are involved with child welfare has not been well explored. Accessing timely treatment is now critical for these parents since the introduction of the Adoption and Safe Families Act of 1997, limiting the time until a permanency decision is made. Using a longitudinal, qualitative approach, substance-abusing parents from 15 families, their relevant family members, and service providers were interviewed approximately every 3 months over an 18-month period. The experiences of these parents add to our knowledge of the unique barriers this population faces, and expands our understanding of the mechanisms by which certain barriers may delay treatment. PMID- 19189806 TI - Parents with co-occurring mental health and substance abuse conditions involved in Child Protection Services: clinical profile and treatment needs. AB - This article reports findings of an exploratory study of 71 parents with substance abuse conditions involved in a child dependency court. Over half (59%) of the parents had a co-occurring mental health condition. Parents with co occurring conditions (PWCC) differed in several important ways from those with only substance abuse conditions. PWCC were also more likely than their case managers were to report a need for mental health treatment. Implications for child welfare practice and research are offered. PMID- 19189807 TI - Perceptions of child neglect among urban American Indian/Alaska native parents. AB - A survey of 101 American Indian/Alaska Native (AIAN) parents in Los Angeles was conducted to explore perceptions of child neglect among urban AIAN parents and factors associated with perceptions. Participants rated substance abuse by parents as the most serious type of neglect. Providing material necessities and providing adequate structure were ranked as the least serious types of neglect. Gender, education, marital status, and indirect experience with Child Protective Services were significantly related to perceptions of neglect among urban AIAN parents. PMID- 19189808 TI - Citizen review panels for Child Protective Services: a national profile. AB - Citizen Review Panels (CRPs) for Child Protective Services are groups of citizen volunteers throughout the United States who are federally mandated to evaluate local and state child protection systems. This study presents a profile of 332 CRP members in 20 states with regards to their demographic information, length of time on the panel, and attitudes regarding the variables that promote and hinder collaboration between the panels and state child welfare agencies. Results indicate that the average review panel member tends to be a professional, middle aged female with an advanced degree. Better communication (between child protective services and the CRPs) and clearer goals/objectives for CRPs were the most cited suggestions of how CRPs and child welfare agencies can work together. Lack of funding and the defensiveness of the child welfare agency were seen as the top obstacles to such collaboration. Policy implications and avenues of further study are discussed. PMID- 19189809 TI - Reasonable efforts? Implementation of the reunification bypass provision of ASFA. AB - The Adoption and Safe Families Act (ASFA) of 1997 includes provisions to deny reunification services under specified conditions and gives states latitude to develop any number of additional "aggravated circumstances" in which parents need not be offered services. California legislators have developed a relatively large number of conditions enabling agencies to bypass reunification services. Based upon a case record review involving 1,055 parents, this study attempts to identify the proportion of parents eligible for a reunification bypass, the proportion recommended to the courts, and the proportion of parents who were denied reunification services, and examines the characteristics of parents associated with reunification bypass recommendations. Based upon focus groups and interviews with child welfare and judicial personnel in six counties, the study also examines the implementation of reunification bypass provisions. Implications for public policy and practice are provided. PMID- 19189811 TI - Multicultural considerations for the application of attachment theory. AB - Attachment styles impact relationships. Research illustrates the importance of attachment from infancy to adulthood across all cultures. The focus of this paper is to explore attachment theory within the context of the therapeutic process with the inclusion of multicultural implications. PMID- 19189810 TI - Using countertransference: analytic contact, projective identification, and transference phantasy states. AB - The influence of projective identification is an integral aspect of most psychoanalytic treatments, not only with patients who are more disturbed, but also with individuals are higher functioning and have neuroses. Projective identification involves both internal relational phantasies of self and object as well as external interactions with the environment. Both elements shape the transference. Continuous projections distort the ego's image of the object, causing introjections that bring increased guilt, anxiety, and envy onto the ego, creating even more radical projections. Consequently, the countertransference is repeatedly stimulated in an evolving or devolving manner (Clarkin, Yeomans, Kernberg, 2006). The case material has illustrated the constant interplay among projective identification, transference, and countertransference as well as the utility of countertransference in making the most helpful interpretations. The concept of analytic contact (Waska, 2006; Waska 2007) was noted as the vehicle of optimal psychological transformation. Rather than an emphasis on frequency, diagnosis, use of couch, or mode of termination, the focus is more on the clinical situation and the moment-to-moment work on internal conflict, unconscious phantasy, destructive defenses, analysis of the transference and extratransference anxieties, and the gradual integration of core object relational experiences. Regarding a more clinical rather than theoretical definition of psychoanalysis, Sandler (1988) states that what truly defines a treatment as psychoanalytic is the analyst's attitudes towards his patient, his willingness to contain and make the effort to patiently understand the patient's unconscious conflicts and reactions to internal phantasy states, the humane detachment and lack of judgment, and the maintenance of a comfortable and safe setting in which the transference can unfold. This definition is certainly similar to the elements of analytic contact. Use of the countertransference is crucial in finding a path into the patient's projective identification processes, which in themselves often hold the core phantasy states with which the patient most struggles. These phantasies shape his experience of the analyst and the world around him. Melanie Klein thought that phantasy was an essential aspect of the mind from birth onwards, with love, hate, and the quest for knowledge being innate, yet influenced by external events that then recast the phantasy and the patient's reaction and perception of those external situations (Spillus, 2007). Klein's belief that phantasy is the bedrock of human experience has led the Kleinian school to conceptualize the transference as being constant and all pervasive, and therefore, not something that arises at only selected times in the treatment. There is no differentiation between the transference and the working alliance. If the analyst is consistently on the alert for manifestations of the transference, he or she may be able to help the patient identify and work through phantasy conflicts. However, the transference, through a constant phenomenon, may elude the analyst for many reasons. Projective identification dynamics are often the culprit because transference phantasies are hidden, discharged, communicated, acted out, camouflaged, or traded off in projective identification interactions with the analyst. It is here that the countertransference, a likewise constant element in the treatment setting, can be most useful in relocating the transference phantasies. The analyst can begin to explore them, rather than be a part of the patient's denial, destructive acting out, or gratifications. As the countertransference is better understood, the analyst can begin to more properly contain, translate, and ultimately interpret the core phantasy states to help the patient feel more in control of them. It is this ongoing emphasis in working within the transference/projective identification/countertransference matrix that gradually brings clarity to the analyst who can share that knowledge as an interpretive proposal. Then, the patient may use it to build greater psychological freedom, emotional stability, and personal choice. PMID- 19189812 TI - Psychotherapy with HIV/AIDS patients: assessment and treatment plan development. AB - Successful psychotherapy with patients who test positive for HIV/AIDS involves a sensitive, careful, and thorough biopsychosocial assessment with specific, detailed attention to the impact that a diagnosis of HIV/AIDS has on overall level of patient functioning. Careful evaluation and review of a patient's level of functioning in the areas of mental health and/or substance abuse, intimate relationships, family relationships, social support and peer relationships, work, and overall health are critical to an accurate assessment. Once the clinician fully evaluates these areas, a treatment plan that identifies specific problem areas can be developed. Clear goals, objectives, and treatment modalities, which address areas of concern, can then be established. PMID- 19189813 TI - The nothing that is something: core dysphoria as the central feature of borderline personality disorder. Implications for treatment. AB - People with borderline personality disorder (BPD) often describe a longstanding depression or dysphoria central to the disorder. The dimension of chronic inner dysphoria is reflected in the DSM criterion of "emptiness" and is demonstrated on self-report measures of depression. In this paper, we report on outcomes for patients with BPD treated with outpatient psychodynamic psychotherapy based on the Conversational Model. Data were collected using the Zung depression scale and the Westmead severity scale at 1 year and at 2 years from two groups of patients. One group had 1 year in psychotherapy, the other had 2 years in therapy. While general symptom scores improved steadily over time in the two-year group, gains for the one-year group tapered off in the second year. Change in depression scores continued at an unchanged rate in the second year for the two-year group compared to a leveling out of gains in the group treated for one year. Gains in general symptom scores were more substantial, suggesting that change in affect occurs more slowly than other symptomatic changes. Drawing upon our findings and those of others in the field, it appears that this slowness of affective change may reflect that patients with BPD have an inner world where predominant negative affectivity became established early in life in the context of disturbances of relatedness. This has implications for the appropriate duration of psychotherapeutic engagement with this group. We argue for more prolonged psychotherapy in this group, which is consistent with current understanding of processes of attachment and development. PMID- 19189814 TI - Teaching the art of empathic interviewing to third-year medical students using a fairy tale--"The prince who turned into a rooster". AB - Can empathy be taught? How can we protect the embryonic forms of empathy germinating in our medical students? Can we immunize them against the ravages to their humanism, astutely observed to occur by Henry Silver, Dean of the University of Colorado in 1982, when he published his clear-sighted commentary, Medical Students and Medical School (Silver, 1982; Krugman, 2008). Although studies show that empathy is damaged during medical school, the author proposes that empathic growth through medical school might be possible if enlightened teaching methods are implemented by governing boards, such as the Association of American Medical Colleges (AAMC), The Liaison Committee on Medical Education (LCME), and the Accreditation Council for Graduate Medical Education (ACGME). The author shares a novel teaching method adapted from a technique used by child psychiatrists, storytelling. PMID- 19189815 TI - Achieving universal immunization in India: the unmet challenge. PMID- 19189816 TI - Work capacity and surgical output for cataract in the national capital region of Delhi and neighbouring districts of north India. AB - OBJECTIVES: To ascertain time taken for cataract surgery by ophthalmologists in the National Capital Region of Delhi and neighbouring districts, to determine what work output is feasible with the available ophthalmologists. METHODS: The time-motion study was conducted during January to June 2006 in the National Capital Region of Delhi and neighbouring districts in North India. Data was collected by observing all activities from entry of a patient into the operating theatre to exit. A total of 156 cataract surgeries performed by 45 ophthalmologists in 38 hospitals were observed. A stop watch was used to record activity time, rounded off to the nearest 10 seconds. Case duration, surgical and clinical times were calculated. RESULTS: Ninety percent ophthalmologists completed surgery in 41.3 minutes. The 10th and 90th percentile for case duration time was 15.5 and 78.4 minutes respectively. Median surgical time was lowest for ophthalmologists working in the NGO sector (10 minutes), compared to the government (23.5 minutes), and private sector (17.3 minutes). Cataract surgical output can be increased in the country if operation theatre time is utilized optimally. PMID- 19189817 TI - Skill building programme in population-based research for medical undergraduates: learners' feedback. AB - OBJECTIVES & METHODS: A training programme in population-based research was introduced for interns in 1993, in a Delhi medical school. Guided by the persistent feedback from learners, the timing of such training was advanced to MBBS para-clinical phase in 1999--integrated with 10 months first slot of community medicine posting. Present article analyzes learners' feedback on this newly designed training programme from 16 consecutive batches from 2002-2006. RESULTS: In 7 of the 9 units of learning, around 90% of the students rated their participatory involvement at > or = 3 points on a 5 point rating scale. This rating was best in data collection (97.8%); and identification of research question (97.6%)--and least in presentation of report (48.7%); and report writing (61.8%). For 77.6% of the students, this was their first hands-on experience in population-based research. Over 55% of the students expressed their willingness to maintain their interest in population-based research after finishing the community medicine posting. On the other hand, 22.6% distinctly expressed their unwillingness in this regard. Main reasons cited by unwilling/not sure students were: 'low level of personal interest in population-based research' (39.7); 'such activity was not contributory in getting admission to postgraduate courses' (33.2%); and 'Not so useful in likely job responsibilities' (21.2%). Almost half (48.9%) of the students chose para-clinical phase as most suitable period of MBBS for such learning exposure. Pre-clinical; clinical; and internship phases were preferred by 19.7%, 13.9%, and 10.7% respectively. CONCLUSION: Present feedback provides us a broad direction in opting for the para-clinical phase where exposure to population-based research can be effectively placed on a systematic basis, without extra resources. PMID- 19189818 TI - Hepatitis E epidemic with bimodal peak in a town of north India. AB - INTRODUCTION: An epidemic of viral hepatitis occurred in Mandi Gobindgarh town of Punjab in northern India during year 2005-06. An attempt was made to study the outbreak clinically, serologically, and etiologically. METHODS: Line listing and spot mapping of all cases of jaundice presented to civil hospital was done. An active search of cases was made through house-to-house visit with the help of 33 teams and 6 supervisors. Twenty two blood samples collected from acute cases were tested for anti-HAV IgM and anti-HEV IgM by ELISA. HEV specific PCR was also carried out. Sanitary survey was also done and water samples were tested for coliforms. RESULTS: In house to house survey 3170 cases of jaundice were reported; of them 2171 (68.5%) were males. Mean age was 28.8 years. Overall attack rate was 5.2%. The epidemic continued for more than a year and bimodal peak was observed. Civil hospital campus which has separate water supply had no jaundice case. About 95% blood samples from icteric patients were found to be positive for IgM and IgG antibodies of HEV. Eighteen persons died during the epidemic, mostly in old age group. Case fatality ratio was 0.57%. No deaths occurred among 17 pregnant women who had developed hepatitis. CONCLUSION: The epidemic was caused by hepatitis E virus, which was transmitted due to faecal contamination of municipal water supply. PMID- 19189819 TI - Public health, preventive & social medicine and community medicine--the name game. PMID- 19189820 TI - A study on human risk factors in non-fatal road traffic accidents at Nagpur. AB - A cross sectional study was conducted among 423 victims of road traffic accidents reporting to Indira Gandhi Medical College, Nagpur for treatment during 1999 2000. Data was collected on pre-designed proforma by interview technique. Majority of the victims were male (85.8%) and of 18-37 years (74%); 64.5% of the subjects were consuming alcohol regularly and 5.9% were drug abusers; 43.7% and 10.2% had visual and hearing impairment respectively; 43.5% were not having any driving experience and 74.4% of the victims with two wheelers were not using any helmets at the time of accidents. Findings highlight the need for sustained health education and enforcement of traffic laws. PMID- 19189821 TI - An ICMR task force study of Prevention of Parent to Child Transmission (PPTCT) service delivery in India. AB - A retrospective survey was conducted during August to December 2007 in 19 medical colleges of India to examine the functioning of the PPTCT service delivery. Data was extracted from records of the PPTCT centers for the year 2005-2006. HIV prevalence was higher than 2005 NACO figures in 11 out of 19 (57.8%) centers. There was wide variation in the proportion of women counseled & tested for HIV in different centers. Antenatal prophylaxis was practiced in 7 out of 19 (36.8%) centers. Overall intra-natal ART was provided to 52.8% of HIV positive women. Early newborn testing was available at 3 out of 19 (15.7%) centers. Improved counseling services are required for better case detection. PMID- 19189822 TI - Nutritional status of Lodha children in a village of Paschim Medinipur district, West Bengal. AB - Undernutrition among tribal children is a major public health problem in India. Our study attempted to evaluate the prevalence of malnutrition among Lodha children of Paschim Medinipur, West Bengal. A cross sectional study was conducted among 165 children aged 1-14 years in a village of Paschim Medinipur district during January to March 2008. Children were considered underweight, stunting and wasting following the NCHS standards. Overall the prevalence of underweight, stunting and wasting was 33.9%, 26.1% and 19.4 %, respectively. Of these, 9.1%, 9.7% and 3.6% children were found to be severely underweight, stunted and wasted. Moreover, the prevalence of underweight and stunting was significantly higher in pre-school children compared to school going children. There is an urgent need for appropriate steps to be taken to improve nutritional status of children in this ethnic group. PMID- 19189823 TI - Prevalence and pattern of childhood morbidity in a tribal area of Maharastra. AB - Previous studies have demonstrated that tribal children suffer from a higher rate of morbidity. Gender discrimination in the form of dietary neglect of the female children has also been noted. The community based cross-sectional study was carried out in tribal PHC Salona of Chikhaldara Block, Amaravati District, Maharashtra to study the prevalence and pattern of morbidities in children. 2603 study children between 0-72 months of age were covered in a house to house survey by the investigator. Parents of eligible children were interviewed using a pre tested questionnaire for socio-demographic details, personal habits, past and current medical history. The prevalence of overall morbidities was 34.7% and it was higher in female as compared to male children (34.8% vs. 29.7%; chi2 = 9.3, p <0.005). Among individual morbidities, the prevalence of acute respiratory infections was the highest (25.5%) followed by acute diarrhoeal diseases (5.8%), conjunctivitis (1.5%), and skin infections (1.2%). PMID- 19189824 TI - A clinico epidemiological study of tetanus cases admitted to epidemic disease hospital, Bangalore. AB - A descriptive study was conducted among tetanus cases admitted to epidemic disease hospital (EDH), Bangalore from October 2006 to March 2007 to describe the socio demographic characteristics of tetanus cases, the clinical presentation and the treatment given to tetanus cases. A total of 80 cases were admitted during this period. Out of the 80 tetanus cases, 71% were from rural area, 55% unimmunized, 47.5% were agricultural workers. A total of 25 (31.25%) deaths were observed during the study period. No case was administered anti tetanus serum outside the epidemic disease hospital. PMID- 19189825 TI - Study of parental handling patterns in a primary school of Kolkata. AB - The intricate pattern of parenting in our socio-cultural context needs evaluation hence this study was done with the objective to reveal the parenting styles of school going children. 141 school children belonging to the age group 6-10 years were included in the study. The Parental Handling Questionnaire a 14-item scale consisting of 10 items of care and 4-items of control was administered to the mothers accompanying the children. Rating was done on a 3-point. The scale showed moderate internal consistency. 5 independent factors with overlapping items of care and control were deduced on factor analysis indicating multidimensional aspects of parenting in our society. PMID- 19189826 TI - Physical wife abuse in an urban slum of Pune, Maharastra. AB - The present cross-sectional study was undertaken during August-September 2004 in an urban slum area of Pune to find out prevalence & characteristics of physical wife abuse/wife beating & to ascertain women's views & behavior towards it in a social context. The prevalence of physical wife abuse/wife beating was reported to be 61.5%. The most commonly reported husband's behavior included slapping (98.8%) followed by pushing (39.8%) & kicking wives (33.7%). 17.8% women justified physical abuse by husband. The most commonly reported behavior of women included crying (85.2%) & silently bearing the physical abuse (59%). PMID- 19189827 TI - Gender inequality in nutritional status among under five children in a village in Hooghly district, West Bengal. AB - A community based, cross-sectional study was conducted in the Mollasimla village of Hooghly district of West Bengal, to examine the differences in nutritional status of under-five males and females and to determine the different bio-social factors associated with such differences. It was found that 55.9%, 51.4% and 42.3% of the girls were underweight, stunted and wasted respectively compared to 46.6%, 40.5% and 35.3% of the boys and a significantly higher proportion of malnutrition was found to be present among female children of higher birth order and those belonging to families with lower per capita income compared to the males. PMID- 19189828 TI - Under nutrition and measles related complications in an outbreak of measles. AB - An outbreak of measles was reported from Shivpuri district of Madhya Pradesh, India. The authors investigated this outbreak with an objective to describe the under nutrition and measles related complications. A total of 723 children aged 12-59 months were included in this study. Their caregiver was interviewed for necessary information and Mid Upper Arm Circumference measurement of the affected child was done for the assessment of nutritional status. 171 (22.3%) of these had reported to have suffered from one or more measles related complication. 556 (76%) of the measles cases in this study were undernourished. Measles related complications was more among measles affected children who were severely undernourished (38%) in comparison to other groups (chi2 = 97.80; d.f. = 1; p<0.0001). PMID- 19189829 TI - Public health strategies to stem the tide of chronic kidney disease in India. AB - It is estimated that 10-15% of the population has chronic kidney disease (CKD), resulting in significant health expenditure, which is largely met by out-of pocket by the patient in India. However, the seriousness of this public health problem has remained largely under-recognized so far. Luckily the preventive measures are simple and not difficult to implement. Public health strategies are essential to control the burgeoning problem. Lifestyle modifications can reduce the incidence of obesity, hypertension and diabetes. These diseases account for a significant proportion of CKD cases. Active involvement of the primary care physicians is vital for early detection of CKD with retardation of its progress, since nephrologists do not see the patients in the early stages. The role of community health specialists is essential to organize health education programs and screening camps, form active patient support groups; and incorporate the prevention program in the various tiers of the health-care system. Availability of optimal nephrology services in peripheral hospitals can rehabilitate most cases of end-stage renal disease, and also prevent the illegal organ trade, which keeps rearing its ugly head at regular intervals in India. PMID- 19189830 TI - Patient's awareness and some behavioural issues related to TB and DOTS. PMID- 19189831 TI - Injury: the most underappreciated and unattended pandemic. PMID- 19189832 TI - Prevalence of risk factors for non-communicable disease in a rural area of Faridabad district of Haryana. AB - BACKGROUND & OBJECTIVES: To estimate the prevalence and levels of common risk factors for noncommunicable disease in a rural population of Haryana. METHODS: The study involved a survey of 1359 male and 1469 female respondents, aged 15-64 years. Multistage sampling was used for recruitment (PHCs/sub-centres/villages). All households in the selected villages were covered, with one male and one female interviewed in alternate household. WHO STEP-wise tool was used as the study instrument which included behavioural risk factor questionnaire and physical measurements of height, weight, waist circumference and blood pressure. The age adjusting was done using rural Faridabad data from Census 2001. RESULTS: The age adjusted prevalence of daily smoked tobacco was 41% for men and 13% for women. Daily smokeless tobacco use was 7.1% and 1.2% for men and women respectively. The prevalence of current alcohol consumption was 24.6% among men and none of the women reported consuming alcohol. The mean number of servings of fruits and vegetables per day was 3.7 for men and 2.7 for women. The percentage of people undertaking at least 150 minutes of physical activity in a week was 77.8% for men and 54.5% for women. Among men 9.0% had BMI > or = 25.0 compared to 15.2% among women. The prevalence of measured hypertension, i.e. > or = 140 SBP and/or > or = 90 DBP or on antihypertensive drugs was 10.7% among men and 7.9% among women. CONCLUSION: The study showed a high burden of tobacco use and alcohol use among men, inactivity and overweight among women and low fruit and vegetable consumption among both sexes in rural areas. PMID- 19189833 TI - Epidemiology of disability in a rural community of Karnataka. AB - OBJECTIVES: To determine the prevalence and pattern of disability in all age groups in a rural community of Karnataka. METHODS: A community-based cross sectional study was conducted during January-December 2004 among 1000 study subjects of all age groups selected randomly from four villages under rural field practice area of a teaching institution. Subjects were interviewed and examined using a predesigned schedule. Percentage prevalence, chi square test and multiple logistic regression analysis were used for statistical analysis. RESULTS: The prevalence of disability was found to be 6.3%. Both physical and mental disabilities are of great concern in this area. 80% of the disabled had multiple disabilities. Knowledge and occupation plays a major role as determinants of disability. Chronic medical conditions are also more common among disabled. PMID- 19189834 TI - Elimination of iodine deficiency disorders--current status in Purba Medinipur district of West Bengal, India. AB - BACKGROUND AND OBJECTIVES: Towards sustainable elimination of iodine deficiency disorders (IDD), the existing programme needs to be monitored through recommended methods and indicators. Thus, we conducted the study to assess the current status of IDD in Purba Medinipur district, West Bengal. METHODS: It was a community based cross-sectional study; undertaken from October 2006-April 2007. 2400 school children, aged 8-10 years were selected by '30 cluster' sampling technique. Indicators recommended by the WHO/UNICEF/ICCIDD were used. Subjects were clinically examined by standard palpation technique for goitre, urinary iodine excretion was estimated by wet digestion method and salt samples were tested by spot iodine testing kit. RESULTS: The total goitre rate (TGR) was 19.7% (95% CI = 18.1-21.3 %) with grade I and grade II (visible goitre) being 16.7% and 3% respectively. Goitre prevalence did not differ by age but significant difference was observed in respect of sex. Median urinary iodine excretion level was 11.5 mcg/dL and none had value less than 5 mcg/dL. Only 50.4% of the salt samples tested were adequately iodised (> or = 15 ppm). CONCLUSION: The district is in a phase of transition from iodine deficiency to iodine sufficiency as evident from the high goitre prevalence (19.7%) and median urinary iodine excretion (11.5 mcg/dL) within optimum limit. But, salt iodisation level far below the recommended goal highlights the need for intensified efforts towards successful transition. PMID- 19189835 TI - Integrated Diseases Surveillance Project (IDSP) through a consultant's lens. AB - India has long experienced one of the highest burdens of infectious diseases in the world, fueled by factors including a large population, high poverty levels, poor sanitation, and problems with access to health care and preventive services. It has traditionally been difficult to monitor disease burden and trends in India, even more difficult to detect, diagnose, and control outbreaks until they had become quite large. In an effort to improve the surveillance and response infrastructure in the country, in November 2004 the Integrated Disease Surveillance Project (IDSP) was initiated with funding from the World Bank. Given the surveillance challenges in India, the project seeks to accomplish its goals through, having a small list of priority conditions, many of which are syndrome based at community and sub center level and easily recognizable at the out patients and inpatients care of facilities at lowest levels of the health care system, a simplified battery of laboratory tests and rapid test kits, and reporting of largely aggregate data rather than individual case reporting. The project also includes activities that are relatively high technology, such as computerization, electronic data transmission, and video conferencing links for communication and training. The project is planned to be implemented all over the country in a phased manner with a stress on 14 focus states for intensive follow up to demonstrate successful implementation of IDSP. The National Institute of Communicable Diseases chosen to provide national leadership may have to immediately address five issues. First, promote surveillance through major hospitals (both in public and private sector) and active surveillance through health system staff and community, second, build capacity for data collation, analysis, interpretation to recognize warning signal of outbreak, and institute public health action, third, develop a system which allows availability of quality test kits at district and state laboratories and/or culture facilities at identified laboratories and a national training program to build capacities for performing testing and obtaining high quality results, fourth, there must be a process established by which an appropriate quality assurance program can be implemented and fifth, encourage use of IT infrastructure for data transmission, analysis, routine communication (E-mail etc) and videoconferencing for troubleshooting, consultations and epidemiological investigations. These five activities must be addressed at the national level and cannot be left up to individual states/districts. PMID- 19189836 TI - Hypertension and epidemiological factors among tribal labour population in Gujarat. AB - A cross sectional study was carried out in 2005 to find out the magnitude of hypertension among 154 tribal labourers of Gujarat belonging to Naika, Rathwa and Damor tribes. WHO classification of hypertension was taken as operational criteria and data was collected in pre-designed, pretested schedule. Blood pressure measurement was done twice on each subject using mercury sphygmomanometer. Overall magnitude of hypertension was found to be 16.9%, and only smoking was found to have significantly associated with it. PMID- 19189837 TI - Respiratory morbidity among street sweepers working at Hanumannagar Zone of Nagpur Municipal Corporation, Maharashtra. AB - Due to the occupational exposure street sweepers are very much vulnerable to develop the chronic diseases of respiratory system. Therefore this study was undertaken to find out the proportion of chronic respiratory morbidity among the street sweepers and the role of various associated risk factors. The study included two groups: study group i.e. street sweepers and comparison group (Class IV workers working in the office buildings). Various risk factors studied were age, sex, socioeconomic status, length of service, smoking habit, type of house, area of residence, cooking fuel and pets. Proportion of chronic respiratory morbidity (chronic bronchitis, asthma and bronchiectasis) was higher (8.1%) among street sweepers compared to comparison group (2.1%), the difference being statistically significant. Unconditional multivariate logistic regression revealed that risk of having chronic respiratory morbidity among street sweepers was 4.24 (95% CI of OR = 1.24 to 14.50) times higher than that in the comparison group and the risk increased significantly with increasing length of service (OR = 1.75, 95 % CI = 1.09 to 2.81). PMID- 19189838 TI - Needle sticks injury among nurses involved in patient care: a study in two medical college hospitals of West Bengal. AB - A hospital-based retrospective study on a sample of 228 nurses involved in patient care, in two medical college hospitals of West Bengal, showed that 61.4% of them sustained at least one Needle Stick Injury (NSI) in last 12 months. The risk of such injuries per 1000 nurses per year was found to be 3,280. Out of the most recent injuries among 140 nurses, 92.9% remained unreported to appropriate authorities; in 52.9% events hand gloves were worn by the nurses; only 5% of those nurses received hepatitis B vaccine, 2.1% hepatitis B immunoglobulin and none of them received post exposure prophylaxis for HIV. PMID- 19189839 TI - Dietary profile of sportswomen participating in team games at state/national level. AB - A cross sectional study was conducted toassess dietary profile of 100 Delhi based national / state level sportswomen, aged 18-25 years, participating in team games -volleyball, hockey, football and kabaddi. Mean energy intake was found to be 1471 +/- 479 Kcal. Only 24 percent of the sports women met the recommendations of 60-65 energy percent from carbohydrates and 87 percent were consuming more than 25 energy percent from fat. The mean macronutrients and micronutrient intakes of all the subjects were much lower than the recommendations. Improper food choices were also observed in majority. It becomes necessary to generate awareness among sports personnel regarding proper nutrition practices. PMID- 19189840 TI - Perception regarding quality of services in urban ICDS blocks in Delhi. AB - The good quality of the services is an important determinant for acceptance of a programme in a community. It not only enhances the credibility of a worker at the ground level but also generate the demand for the services. In this paper perception for the quality of the services was assessed through the exit interview of the beneficiaries at the Anganwadi centres (AWCs). 200 beneficiaries were included from 20 AWCs in a period of one and half month. 52.5% respondents were dissatisfied for the services provided from the AWC for one or more reason. The most common reason mentioned was the not easy accessibility of the AWC and less space available at the AWC (68.6%), followed by the poor quality of the food distributed (66.7%) and irregular pre school education (57.1%) from AWCs. PMID- 19189841 TI - A study on delivery and newborn care practices in a rural block of West Bengal. AB - A cross-sectional study was conducted in a rural block of the State of West Bengal to generate area specific data on the proportion of home deliveries and certain newborn care practices prevalent in that area. The study was done through house-to-house survey among 165 mothers who delivered in last six months. 83.6% deliveries were conducted at home and untrained persons attended 36.3% deliveries. Bath within 24 hours of delivery was given to 17.58% newborns. Birthweight was not recorded in 38.18%. High proportion of newborns, 78.5%, was given prelacteal feeding. The health system should urgently address the deficiencies in the delivery and newborn care practices in the study area. PMID- 19189842 TI - Hospitalisation due to infectious and parasitic diseases in District Civil Hospital, Belgaum, Karnataka. AB - To assess the burden of infectious and parasitic diseases on hospital services at District Civil Hospital (DCH) Belgaum, a retrospective study was carried out using discharge records concerning 8506 inpatients due to infectious and parasitic diseases among 95,655 patients admitted for all causes during the reference period 2000-2003. Out of the 21 causes of infectious and parasitic diseases, only 5 contributed maximally towards hospital admission. The most frequent cause was intestinal infections (44.0%) followed by tuberculosis (35.4%). 57.5% of these admissions were from the productive age group of 20-54 years. Tuberculosis is the most important disease in terms of hospital bed days (59.7%). Tuberculosis and intestinal infectious diseases represent more than three-fourth of the overall burden in terms of hospital bed days. PMID- 19189843 TI - Homelessness: a hidden public health problem. AB - Homelessness is a problem, which affects not only the people who are homeless but the whole society. This problem is not well recognized among the public health professionals. This paper attempts to discuss the issues in the context of homelessness starting from the definition used to methodology of estimation of their numbers as well as their health problems and health care needs. There is lack of data on the health problems of homelessness from India. There is no special health or social programmes or services for this subsection of the society. The existing number of shelters is inadequate and as there are multiple barriers, which prevent them to have proper access to the existing health care system. With the changing social and economic scenario, homelessness is likely to increase. We need to recognize homelessness as a public health problem and attempt to target this group for special care in order to promote equity in health system. PMID- 19189844 TI - HIV/AIDS awareness through mass media--the measurement of efforts made in an urban area of India. PMID- 19189845 TI - Undernutrition in 5-10 year olds: experiences from a PHC in Pondicherry. PMID- 19189847 TI - Circulating non-immune IgG complexes in health and disease. AB - IgG molecules possess a well-defined ability to form complexes with various proteins at interaction sites composed of residues of the constant domains. Such non-immune complexes (non-ICs) were recently identified in the circulatory system of healthy people, as well as patients suffering from various pathologies. By forming non-ICs, attached proteins that are harmful to the organism (anaphylatoxins, for example) are removed from the circulation. Non-immune IgG complexes can react simultaneously with two cell receptors-one specific for IgG, and another specific for an associated protein. Such double reactions augment cellular responses. The attachment of a protein to an IgG site may induce structural changes in neighboring areas of IgG molecules. The formation of non ICs helps proteins with low molecular mass to escape glomerular filtration, as well as enzymatic degradation and cell uptake. Non-immune IgG complexes have been found in commercial immune globulin preparations used for the treatment of various diseases. Among the IgG-attached proteins, there are specific disease biomarkers used for clinical diagnostics and understanding disease processes. Therefore, in order to identify potential biomarkers, not only proteins that are free in the liquid phase of serum but also proteins associated with abundant proteins such as IgG must be investigated. PMID- 19189846 TI - Gene profiling studies in the neonatal ovine lung show enhancing effects of VEGF on the immune response. AB - Preterm and young neonates have an increased predisposition to respiratory distress syndrome (RDS) associated with an immature development of lung surfactant. Glucocorticoids (GCs) are the major immunomodulatory agents used to increase lung development and reduce the mortality and morbidity of preterm infants with RDS. However, their safety remains uncertain, and the precise mechanisms by which they improve lung function are unclear. In previous studies, we found that vascular endothelial growth factor (VEGF) enhances the innate immune response by respiratory epithelial cells, causes a monocytic infiltration into the lung, and reduces the severity of infection by respiratory syncytial virus (RSV), a respiratory pathogen known to affect preterm infants at a high prevalence. The purpose of this study is to measure the effects of VEGF administration on local immune responses in neonatal lambs, as the ovine lung is well suited for comparison to the human lung, due to similarities in alveolar development, immune responses, and RSV susceptibility. We hypothesized that VEGF induces the expression of genes necessary for host immune responses. We analyzed global gene expression profiles in the lungs of neonate lambs treated with VEGF by real-time qPCR. We report that VEGF induced the expression of chemokines (IL 8, RANTES, MCP-1), cytokines (IFN-gamma, IL-6, TNF-alpha, GMCSF), Toll-like receptor (TLR)-4, complement family members (C3, CFB, CFH) and collectins (SP-A, SP-D). These results suggest that VEGF can regulate local immune gene expression in vivo and should be further explored as a potential exogenous therapy for various lung diseases. PMID- 19189848 TI - Effective inhibition of replication of infectious bursal disease virus by miRNAs delivered by vectors and targeting the VP2 gene. AB - RNA interference (RNAi) is a potent mechanism against a variety of viral infections. Infectious bursal disease virus (IBDV) causes an important disease economically in chickens, which is difficult to control. As part of the development of viral vector-mediated RNAi strategy against the disease, five anti VP2 small interference RNAs were selected for construction of microRNA (miRNA) expression vectors tailored for avian cells. Transfection of DF-1 cells with the five vectors resulted in significant inhibition of VP2-EGFP reporter gene expression. More effective miVP2A and miVP2E were selected for further study using single or double miRNA expression vectors. After demonstration of specific miRNA expression, the gene silencing effects were determined in the vector transfected and IBDV-infected cells. Reverse transcriptase PCR and virus titration showed inhibition rates from 76 to 82% on VP2 expression and significant decreases in virus titer by individual and co-expressed miVP2A and miVP2E. The inhibitory effects lasted for at least 120 h after infection with IBDV. These data suggest that the miRNAs targeting the VP2 can inhibit efficiently replication of IBDV. PMID- 19189849 TI - Delivery of a foreign epitope by sharing amino acid residues with the carrier matrix. AB - A broad range of structural viral proteins has the ability to assemble into virus like particles (VLPs). Under the condition that modified subunits are still competent to assemble into VLPs, they are epitope delivery platforms suitable for vaccination purposes. The insertion of foreign sequences can be detrimental for the formation of chimeric VLPs as a result of misfolded subunit proteins. Hence, a strategy was adopted to screen for locations allowing the use of shared residues between the wildtype subunit sequence and the foreign insert. The insertion of a cysteine-containing sequence of hepatitis C virus (HCV) envelope protein 2 (E2) without adding an additional cysteine residue retained the ability of recombinant small hepatitis B surface antigen (HBsAg-S) to form secretion competent VLPs. A cysteine residue shared by the insert and the template protein avoided the formation of non-native disulfide bonds, and allowed the formation of VLPs. The chimeric HBsAg-S VLPs were similar to wildtype VLPs in density exposing the inserted foreign epitope and being immunogenic. Overall, the use of shared sequences between the insert and the subunit will facilitate the design of chimeric VLPs carrying multiple epitopes. PMID- 19189850 TI - A recombinant, infectious human parainfluenza virus type 3 expressing the enhanced green fluorescent protein for use in high-throughput antiviral assays. AB - The ability to rescue an infectious, recombinant, negative-stranded, RNA virus from a complementary DNA (cDNA) clone, has led to new opportunities for measuring viral replication from a viral expressed reporter gene. In this study, the enhanced green fluorescent protein (EGFP) gene was inserted into the human parainfluenza virus type 3 (HPIV-3) antigenome and a recombinant, infectious virus was rescued. Maximum EGFP expression levels, measured by fluorescence, were seen at day 3. Comparison of a 3-day, viral expressed EGFP fluorescence assay to a 7-day, neutral red assay, based on complete cell destruction in virus infected MA-104 cells, yielded Z'-factor values of 0.83 and 0.70, respectively. A 3-day, endpoint EGFP-based antiviral assay and a 7-day, endpoint neutral red based antiviral assay were run in parallel to establish antiviral sensitivity profiles of 23 compounds based on selective index (SI) values. Using an SI threshold of 10, the EGFP-based antiviral assay had a sensitivity of 100% and a specificity of 54%. Thus, the use of an EGFP-based antiviral assay for testing potential antiviral compounds against HPIV-3 in a high-throughput format may be justified. PMID- 19189851 TI - Retroviral self-inactivation in the mouse vagina induced by short DNA. AB - Human immunodeficiency virus (HIV) has been shown to undergo self-destruction upon treatment of cell-free virions with partially double-stranded oligodeoxynucleotides targeting the polypurine tract (PPT) of the viral RNA in the virus particle. The ODN forms a local hybrid with the PPT activating the viral RNase H to prematurely cleave the genomic RNA. Here we are describing the self-destruction of a recombinant lentivirus harboring the PPT of HIV in a mouse vagina model. We showed a decrease in viral RNA levels in cell-free virus particles and a reduction reverse transcribed complementary DNA (cDNA) in virus infected human and primary murine cells by incubation with ODNs. In the vagina simultaneous, prophylactic or therapeutic ODN treatments led to a significant reduction in viral RNA levels. Our finding may have some relevance for the design of other viral self-destruction approaches. It may lead to a microbicide for reduction of sexual and mother-to-child transmission. PMID- 19189852 TI - Global status of tospovirus epidemics in diverse cropping systems: successes achieved and challenges ahead. AB - The diseases caused by thrips-transmitted tospoviruses (genus Tospovirus, family Bunyaviridae) are a major constraint to production of important vegetable, legume and ornamental crops in different parts of the world. Tospoviruses are characterized by having tripartite RNA genomes and utilizing both negative and ambisense genome expression strategies. Their often wide and overlapping host ranges, emergence of resistance-breaking strains, circulative and propagative relationship with polyphagous thrips vectors, and difficulties in predicting their outbreaks pose challenges to development and implementation of effective management programmes. Despite these challenges, for a few tospoviruses, considerable progress has been made in successful development and deployment of practical and effective integrated disease management programmes. This has been due to increased understanding of their molecular biology, plant-virus and virus vector interactions and epidemiology, and to identification of risk factors that contribute to increased disease incidence and of tactics to mitigate those risk factors. However, challenges remain as resistance-breaking or other new strains of known tospoviruses and completely new tospovirus species continue to be described from various parts of the world and have the potential to cause damaging epidemics. To protect crops from the losses caused by severe tospovirus outbreaks, continued vigilance is required to identify and characterize these emerging tospoviruses, determine their impact on crop production, understand their epidemiologies and develop, evaluate and implement control measures to reduce their impact on crop production. PMID- 19189853 TI - PKR protein kinase is activated by hepatitis C virus and inhibits viral replication through translational control. AB - Hepatitis C virus (HCV) infection is currently treated with IFNalpha-based therapy but little is known how IFNalpha inhibits HCV replication. We show here that HCV JFH1 infection of human hepatoma Huh-7 cells leads to the activation of IFN-inducible protein kinase PKR and phosphorylation of the translation initiation factor eIF2alpha. Compared to a control cell HCV replication was significantly elevated in a PKR-knockdown cell, giving rise to a 10-fold higher viral titer, and was less sensitive to IFNalpha treatment. Conversely, transient expression of PKR inhibited HCV replication in a kinase-dependent manner with concomitant increase of eIF2alpha phosphorylation. Further, expression of a phospho-mimetic eIF2alpha mutant moderately inhibited HCV replication. Together, these results demonstrate that PKR is activated by HCV infection and plays a critical antiviral role through inhibition of viral protein translation. PMID- 19189854 TI - Protein-protein interactions in two potyviruses using the yeast two-hybrid system. AB - Interactions between all ten mature proteins of the potyviruses Soybean mosaic virus (Pinellia ternata isolate) and Shallot yellow stripe virus were investigated using yeast two-hybrid (Y2H) assays. Consistently strong self interactions were found between the pairs of HC-Pro, VPg, NIa-Pro, NIb and CP in both viruses. Apart from the NIb, such interactions have been previously reported for some other potyviruses. The 6K1/NIa-Pro combination gave a consistently moderate to strong interaction in both directions for both viruses. This interaction occurred even when the 6K1 of SMV-P was truncated to eliminate the C terminal motif that acts as a recognition site for cleavage by the NIa-Pro. Many other interactions occurred only in one direction or only for one of the two viruses. When taken together with other published reports, the data suggest that interactions detected by Y2H should be regarded as only preliminary indications. PMID- 19189855 TI - In vitro characterization of the Meq proteins of Marek's disease virus vaccine strain CVI988. AB - Gallid herpesvirus 2 (GaHV-2), commonly known as Marek's disease virus serotype-1 (MDV-1), causes T cell lymphomas in chickens. Vaccines prepared from the attenuated CVI988/Rispens MDV-1 strain currently offer the best protection. Although attenuated CVI988/Rispens is non-oncogenic, it codes for at least two forms of the MDV oncoprotein Meq, and these proteins (CVI-Meq and CVI-LMeq) have not been fully characterized. Here, we report that both CVI-Meq proteins, like the Meq protein of Md5 (a very virulent oncogenic strain), were capable of transforming Rat-2 and NIH3T3 cells. Both CVI-Meq and CVI-LMeq proteins activated the meq promoter only in the presence of chicken c-Jun (CK-Jun) whereas Md5-Meq activated the same promoter irrespective of CK-Jun co-expression. However, Meq proteins of both Md5 and CVI988 bound the meq promoter in a ChIP assay regardless of whether CK-Jun was co-expressed. To understand the role of Meq DNA binding and transactivation/repression domains in transcription, we constructed three chimeric Meq proteins, namely, Md5-CVI-Meq, CVI-Md5-Meq, and Md5-CVI-L by exchanging domains between Md5 meq and CVI meq genes. Although these chimeric Meq proteins, unlike CVI-Meq proteins, transactivated the meq promoter, the activation was significantly less than Md5-Meq. To determine the role of individual amino acids, point mutations were introduced corresponding to the amino acid changes of CVI-Meq into Md5-Meq. Amino acid residues at positions 71 and 320 of the Md5-Meq protein were found to be important for transactivation of the meq promoter. All three Meq proteins activated the MDV gB, MMP-3 and Bcl-2 promoters and suppressed transcription from the MDV pp38/pp14 bidirectional promoter. Although no significant differences were observed, decreased transactivation activity was observed with CVI-Meq proteins when compared to Md5 Meq. Collectively, the data presented here indicate that CVI-Meq proteins are generally weak transactivators, which might contribute to the non-oncogenic phenotype of CVI988 virus in chickens. PMID- 19189856 TI - Thermostable foot-and-mouth disease virus as a vaccine candidate for endemic countries: a perspective. PMID- 19189857 TI - HIV vaccine. A short and concise query. PMID- 19189858 TI - Antipsychotic and antidepressant co-treatment: effects on transcripts of inducible postsynaptic density genes possibly implicated in behavioural disorders. AB - Selective serotonin reuptake inhibitors (SSRIs) and antipsychotics co administration is a widely used strategy to treat both psychotic depression and depressive symptoms in schizophrenia. Nonetheless, the molecular mechanisms involved in the therapeutic benefits of antidepressant-antipsychotic combination are still elusive. It has been suggested that co-administration of SSRIs and antipsychotics may result in molecular changes different from their individual effects. In the present study, we evaluated the acute effects of two SSRIs, citalopram and escitalopram, alone or in combination with haloperidol, on the expression of Homer1a together with its splice variant ania-3, and p11, two genes linked respectively to dopaminergic and serotonergic neurotransmission and involved in synaptic plasticity. Homer1a and ania-3 were induced in the striatum by haloperidol, alone and in combination with SSRIs, but not by SSRIs only. Haloperidol+citalopram co-administration induced a stronger Homer1a expression than haloperidol alone in the ventrolateral caudate-putamen. No signal was detected for p11 in striatum, while there were no significant differences among treatments in cortical subregions. Homer1a was significantly down-regulated in the parietal cortex by all treatments. These results demonstrated that haloperidol+citalopram combination exerts synergistic effects on Homer expression, suggesting that citalopram may influence the impact by haloperidol on the dopaminergic neurotransmission. Moreover, present findings confirm that Homer1a and ania-3 are strongly induced in striatum by haloperidol, while they are not influenced by citalopram or escitalopram in this region. Oppositely, in the cortex the two transcripts are modulated by both haloperidol and SSRIs, suggesting a possible role of both dopamine and serotonin in their cortical regulation. PMID- 19189859 TI - Interleukin 8 -251T>A and Interferon gamma +874A>T polymorphism: potential predictors of allograft outcome in renal transplant recipients from north India. AB - BACKGROUND: Episodes of acute rejection represent an important risk factor for the development of chronic allograft nephropathy. We explored whether certain cytokine gene polymorphisms in renal transplant recipients may be useful markers for susceptibility to allograft rejection. Interleukin 8 (IL8) -251 T>A and interferon gamma (IFNG) +874A>T gene polymorphisms were correlated with allograft outcome in renal transplant recipients. METHODS: Genotyping was done by amplification refractory mutational system-polymerase chain reaction (ARMS-PCR) in 264 healthy controls and 296 renal transplant recipients categorized into 235 non-rejecters and 61 rejecters. RESULTS: IL8 -251AA genotype was associated with 2.7-folds increased risk for allograft rejection in recipients experiencing rejection episodes as compared to non-rejecters (OR=2.70, P=0.032). Cox proportional analysis revealed >2-folds increased susceptibility for allograft rejection (HR=2.38, P=0.010) in IL8 -251AA recipients. Kaplan-Meier analysis also demonstrated lower mean time to first rejection episode for IL8 -251AA recipients (23 months) as compared to TT recipients (30 months) (log rank P=0.022). No association of IFNG +874A>T was observed with allograft rejection, however, an increasing trend towards immunosuppressant toxicity was observed in patients with +874TT genotype at one month post transplantation. CONCLUSION: Thus, IL8 -251AA genotype may serve as potential predictor of allograft outcome in our North Indian cohort of renal transplant recipients. PMID- 19189860 TI - Purification and characterization of recombinant human soluble guanylate cyclase produced from baculovirus-infected insect cells. AB - Soluble guanylate cyclase (sGC) has been purified from 100 L cell culture infected by baculovirus using the newer and highly effective titerless infected cells preservation and scale-up (TIPS) method. Successive passage of the enzyme through DEAE, Ni(2+)-NTA, and POROS Q columns obtained approximately 100mg of protein. The sGC obtained by this procedure was already about 90% pure and suitable for various studies which include high throughput screening (HTS) and hit follow-up. However, in order to obtain enzyme of greater homogeneity and purity for crystallographic and high precision spectroscopic and kinetic studies of sGC with select stimulators, the sGC solution after the POROS Q step was further purified by GTP-agarose affinity chromatography. This additional step led to the generation of 26 mg of enzyme that was about 99% pure. This highly pure and active enzyme exhibited a M(r)=144,933 by static light scattering supportive of a dimeric structure. It migrated as a two-band protein, each of equal intensity, on SDS-PAGE corresponding to the alpha (M(r) approximately 77,000) and beta (M(r) approximately 70,000) sGC subunits. It showed an A(430)/A(280)=1.01, indicating one heme per heterodimer, and a maximum of the Soret band at 430 nm indicative of a penta-coordinated ferrous heme with a histidine as the axial ligand. The Soret band shifted to 398 nm in the presence of an NO donor as expected for the formation of a penta-coordinated nitrosyl-heme complex. Non stimulated sGC had k(cat)/K(m)=1.7 x 10(-3)s(-1)microM(-1) that increased to 5.8 x 10(-1)s(-1)microM(-1) upon stimulation with an NO donor which represents a 340 fold increase due to stimulation. The novel combination of using the TIPS method for co-expression of a heterodimeric heme-containing enzyme, along with the application of a reproducible ligand affinity purification method, has enabled us to obtain recombinant human sGC of both the quality and quantity needed to study structure-function relationships. PMID- 19189861 TI - Topiramate-induced erectile dysfunction. AB - Described here is a case of topiramate-induced reversible erectile dysfunction in which possible pathogenetic mechanisms were excluded by use of appropriate psychological, neurophysiological, ultrasound, and laboratory tests. PMID- 19189863 TI - The shiitake mushroom-derived immuno-stimulant lentinan protects against murine malaria blood-stage infection by evoking adaptive immune-responses. AB - Lentinan, a (1-3)-beta glucan from Lentinus edodes, is an effective immunostimulatory drug. We tested the effects of lentinan during blood-stage infection by Plasmodium yoelii 17XL (P.y17XL). Pre-treatment of mice with lentinan significantly decreased the parasitemia and increased their survival after infection. Enhanced IL-12, IFN-gamma and NO production induced by lentinan in spleen cells of infected mice revealed that the Th1 immune response was stimulated against malaria infection. In vitro and in vivo, lentinan can result in enhanced expression of MHC II, CD80/CD86, and Toll-like receptors (TLR2/TLR4), and increased production of IL-12 in spleen dendritic cells (DCs) co-cultured with parasitized red blood cells (pRBCs). Moreover, both the number of CD4(+)CD25(+) regulatory T cells (Tregs) and the levels of IL-10 secreted by Tregs were reduced by pre-treatment with lentinan in the spleen of malaria infected mice. Meanwhile, apoptosis of CD4(+) T cell in spleens of mice pretreated with lentinan was significantly reduced. In summary, lentinan can induce protective Th1 immune responses to control the proliferation of malaria parasites during the blood-stage of P.y17XL infection by stimulating maturation of DCs to inhibit negative regulation of the Th1 immune response by Tregs. Taken together, our findings suggest that lentinan has prophylactic potential for the treatment of malaria. PMID- 19189862 TI - Seizures in the elderly: impact on mental status, mood, and sleep. AB - Comorbidities of epilepsy have not been well explored in the elderly. Herein, we examined mental status, mood, and sleep in elderly patients with epilepsy, compared with age- and gender-matched community controls without epilepsy from the Einstein Aging Study. Testing included a mental status test, the Blessed Information Memory and Concentration (BIMC) test; Prime-MD Patient Health Questionnaire (PHQ) Depression and Anxiety Modules; and Medical Outcomes Study Sleep Scale. Persons with epilepsy (n=31) had higher mean BIMC scores than controls (n=31, BIMC 6.3 vs.1.2, P<0.0001). Mean PHQ Depression scores were higher for cases than controls, indicating more depressive symptoms (4.2 vs 0.8, P=0.006); six cases (18%) and no controls met screening criteria for depression. Mean PHQ Anxiety scores were also higher for cases than controls (3.7 vs 0.0, P=0.001). Cases had poorer sleep scores in the categories of somnolence (P=0.009) and shortness of breath/headache (P=0.021). Thus, comorbidities of epilepsy in this elderly population included decreased mental status, a higher prevalence of depression and anxiety, and poorer sleep health when compared with agemates without epilepsy. Mental status impairment was not related to antiepileptic medication or mood disturbance. Further investigation will explore these associations prospectively. PMID- 19189864 TI - Effects of 1-bromopropane, a substitute for chlorofluorocarbons, on BDNF expression. AB - 1-Bromopropane (1-BP) has been widely used as an alternative to ozone-depleting chlorofluorocarbons in various industries. Although the neurotoxicity of 1-BP has been recently reported, there is little information about the effect of 1-BP on the cells in brain by experimental approach. Here we studied the effect of 1-BP on brain-derived neurotrophic factor (BDNF) expression in astrocytes in vitro. The BDNF mRNA level was remarkably decreased by 1-BP in a human astrocytoma cell line, U251, and in mouse primary astrocytes. The DNA-binding and specific reporter activity of cAMP response element-binding transcription factor (CREB), which is one of the key molecules regulating BDNF expression, were reduced by 1 BP in U251 and/or mouse primary astrocytes. Additionally, protein kinase A (PKA) activity was suppressed by 1-BP in U251. These results suggest that BDNF expression was affected by 1-BP through at least PKA. PMID- 19189865 TI - Fluoxetine treatment to rats modifies serotonin transporter and cAMP in lymphocytes, CD4+ and CD8+ subpopulations and interleukins 2 and 4. AB - Several lines of evidences indicate that antidepressants produce various immunomodulatory effects. Fluoxetine, an antidepressant and selective serotonin reuptake inhibitor, modulates immune cells in vitro. To explore the in vivo influence of fluoxetine on lymphocytes, male Sprague-Dawley rats were treated daily, 10 mg/kg, or with saline solution for 1, 2 and 3 weeks. The presence of serotonin transporter in CD3+, CD4+ and CD8+ subpopulations of T lymphocytes was determined by immunofluorescence. Serotonin transporter was also labeled with [(3)H]paroxetine, specific binding defined with imipramine. Plasma levels of pro inflammatory interleukin 2 (IL-2), and anti-inflammatory interleukin 4 (IL-4), were measured by ELISA; and cAMP concentration by radioimmunoassay. Fluoxetine significantly increased the number of lymphocytes expressing serotonin transporter and elevated the binding of [(3)H]paroxetine. The percentage of CD4+ cells decreased, that of CD8+ increased, and CD3+ did not change. The ratio CD4+/CD8+ was significantly lowered. Fluoxetine administration elevated the levels of IL-4 at 1, 2 and 3 weeks; and of IL-2, at 2 and 3 weeks. IL-4/IL-2 ratio was significantly increased in fluoxetine group respecting the controls and was similar during the 3 weeks of treatment. Fluoxetine produced a significant decrease in cAMP concentrations in lymphocytes, probably by secondary activation of serotonin receptors. Treatment with fluoxetine modified immune parameters in plasma and lymphocytes of rats, which might be relevant for its systemic therapeutic action as an antidepressant. PMID- 19189866 TI - Apoptosis in pemphigus. AB - Pemphigus is a severe autoimmune blistering skin disease. The two main forms of pemphigus, pemphigus vulgaris and pemphigus foliaceus, are primarily caused by autoantibodies against the desmosomal cadherins desmoglein 1 and desmoglein 3. The histopathological hallmark is the loss of cell adhesion between neighbouring keratinocytes, a phenomenon called acantholysis. Various underlying mechanisms of acantholysis have been described including apoptosis. The role of apoptosis in pemphigus pathogenesis is unclear at present and the focus of this review. While characteristic signs of apoptosis can be found in lesional patient skin these can be missing in very early lesions of macroscopically perilesional skin where acantholysis is detected by light microscopy. Under experimental conditions, activation of apoptotic signalling can be induced by pemphigus IgG as well as by pemphigus serum which contains large amounts of Fas ligand. Studies using caspase inhibitors which were effective to block acantholysis indicate that the apoptotic machinery contributes to cell dissociation. However, acantholysis can occur in the absence of apoptosis which demonstrates that apoptosis is not strictly required for skin blistering in pemphigus. Taken together, signalling events leading to apoptosis including activation of executioner caspases are not required but may participate in the cellular response to pemphigus autoantibodies leading to acantholysis. PMID- 19189867 TI - Interleukin-6 as a key player in systemic inflammation and joint destruction. AB - Interleukin-6 (IL-6) is a cytokine that can facilitate autoimmune phenomena, amplify acute inflammation and promote the evolution into a chronic inflammatory state. In addition, it is a major promoter of bone resorption in pathological conditions. In particular, IL-6 has a pivotal role in synovitis, bone erosions and in the systemic features of inflammation. This cytokine specifically binds to IL-6 receptor (IL-6R), forming the IL-6/IL-6R complex that binds to gp130, a membrane-bound protein, which is involved in non-ligand-binding signal transduction. Targeting IL-6R in both animal models of arthritis and in rheumatoid arthritis patients with a humanized anti IL-6R monoclonal antibody (tocilizumab) effectively controls local and systemic inflammatory manifestations and blocks cartilage and bone destruction. Given the pleiotropic function of IL-6 it can be anticipated that other inflammatory diseases and bone metabolic conditions might benefit from selective IL-6 signaling inhibition. PMID- 19189868 TI - Decreased perinatal mortality among women with diet-controlled gestational diabetes mellitus. AB - OBJECTIVE: To examine pregnancy outcomes associated with diet-controlled gestational diabetes mellitus (GDM A1). METHODS: A retrospective cohort study compared pregnancy characteristics of women with and without GDM A1 at a center where GDM A1 patients are routinely induced at 40 weeks. RESULTS: Higher rates of complications such as shoulder dystocia, congenital malformation, and macrosomia were observed in GDM A1 patients. A lower incidence of perinatal mortality was present in GDM A1 women compared with women without GDM A1. This association lost its significance when controlled for maternal age, ethnicity, induction, cesarean delivery, and birth weight in a multivariate model. Although the stillbirth rate before 40 weeks of gestation was identical among all participants, after 40 weeks it was significantly higher in women without GDM A1. CONCLUSION: Induction of women with GDM A1 at 40 weeks may play a role in lowering perinatal mortality to below that of the general population. PMID- 19189870 TI - Paratubal borderline serous tumors. AB - BACKGROUND: While ovarian borderline tumors are common, their fallopian tube counterparts are extremely rare. We describe the first case of a serous borderline paratubal tumor with a review of the literature on paratubal and tubal low malignant potential tumors. CASE: A 26 year-old woman presented with acute onset of sharp right lower quadrant pain and nausea. Abdominal and pelvic CT revealed 12.5 cm cystic ovarian mass and she was admitted for pain control. On hospital day two, her pain escalated and torsion was suspected; thus, she underwent a diagnostic laparoscopy with right fallopian tube cystectomy and partial salpingectomy. Inspection of the cyst wall demonstrated multiple papillary excrescences and frozen section returned at least borderline fallopian tube neoplasm. After a fertility-sparing comprehensive surgical staging procedure was performed, she was diagnosed with stage IC fallopian tube serous borderline tumor and underwent no further therapy. Currently, she is without evidence of disease recurrence. CONCLUSION: Borderline fallopian tube or paratubal tumors are usually early stage at diagnosis and commonly present in the third decade with pain or are discovered on routine examination. Continued reporting of these tumors is essential to understanding the prognosis and treatment of this rare tumor. PMID- 19189871 TI - The use of lithium, valproate or lamotrigine for psychiatric conditions in children and adolescents in Norway 2004-2007 - a prescription database study. AB - BACKGROUND: There is an increasing awareness of bipolar disorder in children and adolescents. Our study aimed to explore how common the use of lithium, valproate and lamotrigine in psychiatric conditions in children and adolescents in Norway and to describe concomitant use of other psychotropic drugs. METHODS: The Norwegian Prescription Database (NorPD) captures all prescriptions given to individuals in ambulatory care. We investigated prescriptions for lithium, and for valproate and lamotrigine given for psychiatric conditions to children and adolescents under the age of 18 years from 2004 till 2007. We also studied other prescriptions for psychotropic drugs given to the same patients during the period. A comparison was made with the users of ADHD medication. RESULTS: The number below 18 years treated with lithium, valproate or lamotrigine rose from 323 (0.03%) in 2004 till 454 (0.04%) in 2007. Lamotrigine increased most rapidly and lithium was most infrequently used. The users of the three drugs had often also tried antidepressants (14-42%) and atypical antipsychotics (21-51%), but true concomitant use was rarer (7-20% and 16-43% respectively). The users of ADHD medication had a much lower use of other psychotropic drugs, but quite a few users of lithium, valproate and lamotrigine had tried ADHD medication (20-26%) and even used these drugs concomitantly (15-21%). LIMITATIONS: NorPD does not include diagnoses, and we do not know if the drugs dispensed were actually used. CONCLUSION: Overall there has been a slight increase in the use of lamotrigine in Norway during the last four years, most likely prescribed as a mood stabilizer in bipolar disorders. Still the use among the under 14-year-olds is rare in Norway. PMID- 19189872 TI - Autoimmune spread to myelin is associated with experimental autoimmune encephalomyelitis induced by a neuronal protein, beta-synuclein. AB - Accumulating evidence suggests that autoimmunity against neuronal proteins is important for MS pathogenesis. We have characterized T- and B-cell responses associated with experimental autoimmune encephalomyelitis (EAE) induced in Lewis rats with recombinant beta-Synuclein (betaSync), a neuronal component. The encephalitogenic betaSync-specific T cells recognize a single immunodominant region with an epitope delineated at amino acids 97-105; B-cell specificity is more widespread, albeit directed mostly to the C-terminus of betaSync. Most interestingly, betaSync-induced autoimmune T- and B-cell responses spread not only to other neuronal antigens but also to myelin encephalitogens, raising the possibility that anti-neuronal immune attacks could also result in demyelination. PMID- 19189873 TI - Chemoreflex sensitivity and congenital heart disease. PMID- 19189874 TI - Is low adiponectin levels more sensitive predictor of in-stent restenosis: new questions and challenges. AB - Predicting in-stent restenosis has been subject of many investigations. Several candidates have also been proposed such as CRP. However, recent findings suggested that adiponectin may be a more sensitive marker for in-stent restenosis. These initial results should be confirmed by other large scale prospective trials. PMID- 19189875 TI - Dual visualization: a technique to simplify native vessel intervention in patients with bypass grafts. AB - We describe a case of stenting to a distal left anterior descending artery stenosis via the native vessel aided by contrast injection via a left internal mammary artery graft. This dual visualization technique allows good distal vessel opacification whilst minimising instrumentation of the internal mammary artery and its associated risks. PMID- 19189876 TI - Phentermine cardiovascular safety. In response to Yosefy C, Berman M, Beeri R. Cusp tear in bicuspid aortic valve possibly caused by phentermine. International journal of cardiology 2006;106:262-3. PMID- 19189877 TI - Weight growth velocity of very low birth weight infants: role of gender, gestational age and major morbidities. AB - It's well known that VLBWI fail to thrive, however it's still unclear how gender, GA and morbidities affect growth pattern: aim of this study is to assess the influence of these factors on weight growth. 262 VLBWI were selected. Weight was recorded daily up to 28 days, weekly up to discharge and during 7 scheduled follow-up visits up to 2 years of corrected age. Individual profiles were fitted with a mathematical function suitable to model selected growth milestones and mean distance and velocity curves were drawn. Effects of gender, GA, major morbidities, nutritional and respiratory support on individual weight growth milestones were estimated using a multivariate linear model. Each of these variables acts differently on weight growth pattern mainly modifying velocity curves characteristics. In particular, infants with major morbidities weight growth impairment-seen on distance curves at 2 years of corrected age-depends on poor weight velocity during a critical period ending within 4th month of postnatal age, for SGA or BPD infants, starting from 5th month of postnatal for severely neurologically impaired infants. These critical periods could be the most appropriate to identify risk factors for weight growth impairment in VLBWI. PMID- 19189878 TI - Reduction of functional disability with atypical antipsychotic treatment: a randomized long term comparison of ziprasidone and haloperidol. AB - BACKGROUND: Recent interest has focused on the definition and measurement of clinical remission in people with schizophrenia. This study examined the process of development of "functional remission" in a long-term comparative double-blind study of haloperidol and the atypical antipsychotic medication ziprasidone. METHODS: Community dwelling patients with schizophrenia were randomized to treatment with haloperidol (n=47) or ziprasidone dosed either once or twice daily (n=139). They were re-examined at follow-up intervals that ranged up to 196 weeks. Their community functioning was examined with the Heinrichs-Carpenter Quality of Life Scale (QLS). Total scores for occupational and interpersonal functioning and achievement of improvement milestones across the individual items were both analyzed. RESULTS: Mixed model repeated measures analyses detected a significant (p<.05) treatment effect over time favoring ziprasidone for interpersonal functioning. While the mixed model was not significant for role functioning, the mean change at endpoint was significantly greater than 0 for the ziprasidone group but not the haloperidol group. Analyses of the distributions of change scores across the items showed that the number of items where endpoint scores were 5 or 6 (reflecting minimal to no impairment) was significantly higher in ziprasidone treated patients, (p=.03). IMPLICATIONS: Long term treatment with ziprasidone was associated with greater functional gains than treatment with haloperidol, even when the time course of dropout was controlled. Both treatment retention and functional gains favored the atypical treatment in this long-term study. Future long-term studies will be needed to clarify the determinants of these functional changes. PMID- 19189879 TI - Behavioral changes in G72/G30 transgenic mice. AB - Genetic studies have implicated the evolutionary novel, primates-specific gene locus G72/G30 in schizophrenia, bipolar and panic disorders. It encodes for a protein LG72 whose function has been controversially discussed as putative regulator of the peroxisomal enzyme D-amino-acid-oxidase (DAO), or as a mitochondrial protein, which promotes robust mitochondrial fragmentation in mammalian cell lines including human and rat primary neurons. Because of this conserved function we here have generated "humanized" BAC transgenic mice (G72Tg) expressing alternatively spliced G72 and G30 transcripts, and the LG72 protein. G72 expression is prominent in granular cells of the cerebellum, the hippocampus, the cortex and the olfactory bulb. Most strikingly, G72Tg mice displayed deficits in sensorimotor gating which could be reversed with haloperidol, increased sensitivity to PCP, motor-coordination deficits, increased compulsive behaviors and deficits in smell identification. These results demonstrate that expression of the human G72/G30 gene locus in mice produces behavioral phenotypes that are relevant to psychiatric disorders. PMID- 19189881 TI - [Annals of Clinical Biology on the rails]. PMID- 19189880 TI - An epigenetic intervention interacts with genetic strain differences to modulate the stress-induced reduction of flurazepam's antiseizure efficacy in the mouse. AB - Stress induces changes in the endogenous tone of both GABA and NMDA receptor mediated neurotransmission in the intact mouse. Because changes are observed 24 h after stress, epigenetically-regulated alterations in gene expression may mediate these effects. In earlier work, sodium butyrate, a centrally-active histone deacetylase inhibitor that promotes gene expression, was shown to modulate the stress-induced reduction of the ability of MK-801 (dizocilpine), a noncompetitive NMDA receptor antagonist, to antagonize electrically-precipitated seizures. In the current study, we extended this work to look at sodium butyrate's modulatory effect on stress-induced changes in the antiseizure efficacy of flurazepam, a benzodiazepine receptor agonist, in two strains of mice. Epigenetic mechanisms, genetic strain differences and a standard stress interacted to alter flurazepam's antiseizure efficacy. These data support examination and development of epigenetic treatment strategies. PMID- 19189882 TI - [Laboratory assessment of HDL heterogeneity and function]. AB - BACKGROUND: Plasma concentrations of HDL cholesterol (HDL-C) and its major protein component apolipoprotein (Apo) A-I are strongly inversely associated with cardiovascular risk, leading to the concept that therapy to increase HDL-C and ApoA-I concentrations would be antiatherosclerotic and protective against cardiovascular events. The recent failure of the drug torcetrapib, a cholesteryl ester transfer protein inhibitor that substantially increased HDL-C concentrations, has brought focus on the issues of HDL heterogeneity and function as distinct from HDL-C concentrations. CONTENT: This review addresses the current state of knowledge regarding assays of HDL heterogeneity and function and their relationship to cardiovascular disease. HDL is highly heterogeneous, with subfractions that can be identified on the basis of density, size, charge, and protein composition, and the concept that certain subfractions of HDL may be better predictors of cardiovascular risk is attractive. In addition, HDL has been shown to have a variety of functions that may contribute to its cardiovascular protective effects, including promotion of macrophage cholesterol efflux and reverse cholesterol transport and antiinflammatory and nitric oxide-promoting effects. SUMMARY: Robust laboratory assays of HDL subfractions and functions and validation of the usefulness of these assays for predicting cardiovascular risk and assessing response to therapeutic interventions are critically important and of great interest to cardiovascular clinicians and investigators and clinical chemists. PMID- 19189883 TI - [Uranium: properties and biological effects after internal contamination]. AB - Uranium is a radionuclide present in the environment since the origin of the Earth. In addition to natural uranium, recent deposits from industrial or military activities are acknowledged. Uranium's toxicity is due to a combination of its chemical (heavy metal) and radiological properties (emission of ionizing radiations). Acute toxicity induces an important weight loss and signs of renal and cerebral impairment. Alterations of bone growth, modifications of the reproductive system and carcinogenic effects are also often seen. On the contrary, the biological effects of a chronic exposure to low doses are unwell known. However, results from different recent studies suggest that a chronic contamination with low levels of uranium induces subtle but significant levels. Indeed, an internal contamination of rats for several weeks leads to detection of uranium in many cerebral structures, in association with an alteration of short term memory and an increase of anxiety level. Biological effects of uranium on the metabolisms of xenobiotics, steroid hormones and vitamin D were described in the liver, testis and kidneys. These recent scientific data suggest that uranium could participate to increase of health risks linked to environmental pollution. PMID- 19189884 TI - [PSA kinetics after total prostatectomy]. AB - The prostate specific antigen (PSA) is the best marker of the prostate cancer today although not very specific of this pathology. The dynamic interpretation of this marker always has to prevail over that of overtaking a threshold. With the lack of residual cancer, PSA becomes undetectable by the first month after total prostatectomy: less than 0.1 microg/L. The type of diminution mono- or biphasic of the marker depends on the chronology of the takings. Faced with residual cancer, PSA either does not become undetectable or increases after an initial undetectable period. A recurrence is defined by a value of PSA higher than 0.2 microg/L and confirmed on two successive assays. The time of appearance of the recurrence and the PSA doubling time after total prostatectomy have, with the initial clinical stage and the Gleason score, a diagnostic value on the nature of the site of recurrence, local or metastatic. PMID- 19189885 TI - [Evaluation of prescription and interpretation of microalbuminuria by general practitioners]. AB - Microalbuminuria is well recognized as an independent marker of early renal failure in patients with diabetes mellitus and hypertension. We describe here the french results of an international study on the use and interpretation of microalbuminuria by general practitioners. A case history based questionnaire upon a type 2 diabetic patient was sent to 600 general practitioners in the Champagne-Ardenne Region to identify their habits in terms of prescription and of results interpretation. The analysis of the results shows a great variability of practices, regarding the procedures of urine collection, the units used, or the decision limits. These discrepancies can lead to inappropriate care of the patient. Even though national recommendations on the use of MA have been made, this study highlights the necessity for general practitioners to refer to concerted and consensual practices. PMID- 19189886 TI - [Evaluation of Variant II analyzer equipped with the new 270-2101 NU kit (Bio Rad) for HbA 1c assay]. AB - HbA(1c) represents a key parameter in the follow-up of glycemic balance in diabetic patients. It may be assayed by different methods, among which high pressure liquid chromatography (HPLC). We have evaluated a new method available on HPLC Variant II analyzer (BioRad) equipped with the new kit 270-2101 NU. Chromatographic separation is improved, allowing a better identification of peaks. Intra- and inter-assay coefficients of variation are respectively lower than 1.1% and 1.8%. Linearity is excellent from 3.2% to more than 18%. The correlation with the previous method (kit 270-2101) is good: y (% HbA(1c) new kit) = 0.944x (% HbA(1c) previous kit) + 0.299, r(2) = 0.995. There is no inter sample contamination. This method is less sensitive to interferences frequently found in practice (labile glycated hemoglobin, carbamylated haemoglobin) than the previous one. Validation is possible in more circumstances when an abnormal hemoglobin is present (especially in case of hemoglobin D or E). As the control of analytic quality is a major element for validation and clinical use of HbA(1c) results, the characteristics of this new method make it a well-suited tool for daily laboratory practice. PMID- 19189887 TI - [Hematocrit measurement: comparison of conductimetry to microcentrifugation]. AB - In general, blood gas analysers can also determine the value of haematocrite by measuring the blood's conductivity. The question to ask is whether this value is reliable. In this study, hematocrit obtained via conductivity from 6 different pieces of equipment were compared with those measured using the gold standard method, which is microcentrifugation. By interpreting the results of 320 arterial blood samples taken in the intensive care unit DAR "B" we can see that the reliability between two measurements on the same piece of equipment is very good, in general > 0.95 whatever the equipment. The reliability between the means of the two measurements and the gold standard is slightly lower but remains very satisfactory, most often between 0.8 and 0.9. The Gem Premier 3000 (IL) analyser and the Roche OMNI S gave the best reliability compared with centrifugation. The Spearman coefficients between the mean values of the analysers and those of centrifugation were high, with the exception of the Rapidpoint 405. They are all statistically different from zero (p<0.0001). PMID- 19189889 TI - [Improvement of the performances of the messages for the identification of the lymphocyte population with the Beckman Coulter LH750 haematology analyser]. AB - We have compared the efficiency of the detection for lymphocyte abnormalities using the LH 750 analyzer according to standard CLSI recommendations (Clinical and Laboratory Standards Institute) and three new rules dedicated to abnormal lymphocyte recognition. These new rules are defined by combining quantitative and qualitative criteria. They improve the detection of lymphocytes with abnormal morphology with an efficiency of 97.7% as compared to 81% with the standard protocol and a sensitivity of 92.8% compared to 23% initially. Regarding the detection of large granular lymphocytes, the new rules demonstrate an efficiency of 97.6%, a specificity of 81.8% and a specificity of 98.2%. These new criterias, which can be easily implemented in the settings of the automatic counter improve the technical efficiency of the validation process with an expected decrease of the number of manual slide reviews. PMID- 19189890 TI - [Evaluation of HBA1C measurement on Architect CI8200 (Abbott Diagnostic). Comparison with HPLC D-10 Bio-Rad assay]. AB - The aim of this study is to present an evaluation of HbA(1c) Assay on Architect CI8200 (Abbott Diagnostic). The measurement includes Hb assay by colorimetry and HbA(1c) by immunoturbidimetry. The percentage of HbA(1c) is the report HbA(1c)/complete Hb with a conversion coefficient. Repetability (n = 30; CV: 1.15 1.91%) and reproductibility (n = 30; CV: 2.09-2.64%) are good. Abbott results cannot be returned above 12%. Comparison between HbA(1c) Abbott and HbA(1c) Bio Rad is performed on 161 patients samples ranging from 4.7 and 12%. Results show a correlation coefficient of 0.9847 (N = 161) with a regression equation: [Abbott] = 1.02x [Bio-Rad]-0,636]. Differences between two methods are normally distributed. 95% of differences lie between limits (-0.61%; +0.61%). Such differences are clinically important and interchangability of two measurements can't be possible now because lack of agreement. We hope that IFCC standardization will reduce these differences. Presence of a jaundice and carbamylation of haemoglobin do not interfer with Abbott assay. Hemoglobin variants are not detected. Therefore, monitoring of diabetic patients with HbA(1c) is possible only if hemoglobinopathy has been identified before. PMID- 19189891 TI - [Principles recommended to reassure the identification of the patient, the request of analysis and the report of biological results]. AB - The collection of a reliable identity is required for the constitution of any medical file, and biological file in particular. The regulation concerning the rights of the patient, the organization of the Social Security, and the respect of the private life must be applied. The biologist must comply with the GBEA with a special attention when it comes to transfusion. Each individual is unique and should be identified by a single number. Maximum precautions are essential as for the data confidentiality. Adapted procedures (Charter of patient collection of the identity, procedure of bringing together of the identities) with traceability of all the operations are essential but non-sufficient preliminaries. The identitovigilance recommended by the CNIL, must be implemented as well as all other already vigilances required by the law. In this context, the biologist must be particularly active. He must ensure the correct collection of the data, the description of the anomalies and the redundant data, and take into account the corrections. By its transverse position, he plays an important part and must be part of the cells of identitovigilance. It must respect its specific requirements for the corrections of identities and must be informed for any modification taking place after the biological validation. The principal recommendations making it possible to ensure the reliability of the identification of the patient and the biological file were joined together in this article by a working group from the National College of Biochemistry of the Hospitals (CNBH). PMID- 19189892 TI - [System analytical approach of lung function and hemodynamics]. AB - The authors critically analyse the traditional views in physiology and complete them with new statements based on computer model simulations of lung function and of hemodynamics. Conclusions are derived for the clinical practice as follows: the four-dimensional function curves are similar in both systems; there is a "waterfall" zone in the pulmonary blood perfusion; the various time constants of pulmonary regions can modify the blood gas values; pulmonary capillary pressure is equal to pulmonary arterial diastole pressure; heart is not a pressure pump, but a flow source; ventricles are loaded by the input impedance of the arterial systems and not by the total vascular (ohmlike) resistance; optimum heart rate in rest depends on the length of the aorta; this law of heart rate, based on the principle of resonance is valid along the mammalian allometric line; tachycardia decreases the input impedance; using positive end expiratory pressure respirators the blood gas of pulmonary artery should be followed; coronary circulation should be assessed in beat per milliliter, the milliliter per minute may be false. These statements are compared to related references. PMID- 19189893 TI - [Colonoscopy, the primary tool for colonic screening]. AB - Through an overview of the colonic screening methods, the authors study colonoscopy with high emphasis. Taking into consideration its high sensitivity and specificity in the detection of colorectal carcinomas and adenomas, as well as the possibility of adenoma removal through examination, colonoscopy is recommended to be applied as the primary colonic screening within the scope of a national screening program. PMID- 19189894 TI - [Comparison of postoperative complications following bone harvesting from two different donor sites for autologous bone replacement (hip bone and proximal epiphysis of the tibia)]. AB - Whereas autologous bone replacement was earlier applied in maxillofacial surgery virtually only for the restoration of mandibular defects and for the osteoplasty of patients with cleft alveolar process, the free transplantation of autologous bone (spongiosa or cortical bone or both) is nowadays primarily used for implantation purposes. Autologous bone is still the gold standard for bone replacement. This is the case even though a wide selection of bone substitutes is currently available, with which new bone equivalent to autologous bone can be produced in certain cases. Autologous bone is often obtained from intraoral sources, but if a larger quantity of spongiosa is required, these sites (the chin, the retromolar area of the mandible, the muscular process, etc.) are not suitable. Of the extraoral donor sites, the most frequently used site is the iliac crest, but the proximal epiphysis of the tibia is also appropriate for this purpose since we have recently performed bone transplantations on appreciable numbers of patients, we decided to compare the morbidity data relating to the two donor sites. In the 9 months between March and November 2007, sinus elevations were carried out on 14 patients with bone taken from the tibia, while in 38 patients bone was taken from the iliac crest for osteoplasty on clefted alveolar process. The comparison was based on postoperative clinical examinations, the complaints of the patients and objective study of the morbidity relating to the two donor sites. Clinically the patients tolerated both interventions well. Mobilization took place on the day of intervention. There were no major complications; one minor haematoma was observed after each type of surgery. The postoperative complaints of the patients did not reveal any essential difference. Following bone harvesting from the iliac crest, the gait of the patients slightly hampered for up to 10 to 14 days. In the tibia cases, the patients experienced no pain on walking by the second day. As regards donor site morbidity, protracted (1 2 weeks) oedema was observed after hip surgery, with paraesthesia of the area of innervation of the n. cutaneous femoris lateralis in 1 case, while there was a minor seroma following tibia surgery in 1 case. Our clinical experience suggests that, if 10-15 cm(3) spongiosa is required for augmentation purposes and there is no need for cortical bone, the patient is exposed to less stress when bone is taken from the proximal epiphysis of the tibia. PMID- 19189895 TI - [Surgery or invasive endoscopy for the treatment of chronic pancreatitis?]. AB - Chronic pancreatitis is primarily an internal medical disease. However, conservative therapy does not always solve the symptoms, and in such cases some kind of intervention is needed. The related possibilities are examined in the present study. Surgical indications are intractable pain, pseudocyst formation, compression syndrome and the suspicion of malignancy. Invasive endoscopy is also involved in the management of chronic pancreatitis, that could be the first-line therapy in selected cases, namely in the case of pseudocyst, among poor-risk patients for the treatment of stenosis of biliary and pancreatic duct and also in the case of partial rupture of the Wirsung-duct. Further trials are needed to elucidate the problems. It can be concluded that both surgical and endoscopic therapy have place in the armamentarium of the treatment of chronic pancreatitis, based on suitable criteria. PMID- 19189896 TI - [Frequent high-level burnout among general practitioners and residents]. AB - General practitioners (GPs) play a central role in patient care and are exposed to high levels of work strain and consequent burnout due to the large number of stressful patient-doctor relationships. Despite the high likelihood of burnout among GPs, limited information is available about this topic. AIMS: To explore the prevalence of burnout among GPs and residents in Hungary. METHODS: Exploratory/descriptive, cross-sectional study with self-administered questionnaires among 453 GPs and 43 residents. To assess burnout, the Maslach Burnout Inventory (MBI-GS) was used. To evaluate the level of burnout, mean (+/- SD) scores on the emotional exhaustion, cynicism/depersonalization, and personal accomplishment dimensions of the MBI were determined among male and female GPs and residents. Differences in the level or degree of burnout (high, intermediate and low) in all three burnout dimensions between male and female GPs and residents were examined by independent samples t -test and chi 2 -tests. Socio demographic antecedents to burnout were assessed by linear regression analyses. RESULTS: Residents reported significantly lower cynicism/depersonalization [ t (df): 2.8 (476); p < 0.01] and personal accomplishment [ t (df): 2.0 (485); p < 0.05] compared to GPs. No gender differences were identified in the level of burnout. Significantly more GPs then residents reported high [chi 2 (df) = 5.9 (1); p < 0.05] or intermediate [chi 2 (df) = 4.6 (1); p < 0.05] degree of depersonalization. Emotional exhaustion, depersonalization, and low personal accomplishment were reported by around 30%, 60%, and 100% of the physicians, respectively. Being a resident emerged as the strongest negative predictor of depersonalization (beta = -0.09, 95% CI -0.22 - -0.002). CONCLUSIONS: The prevalence of burnout is high among GPs, and almost all GPs report low degree of personal accomplishment. Residency emerged as a significant protective correlate of depersonalization. These findings provide further data for cross-cultural burnout research. PMID- 19189897 TI - [Defensive movements of the fetus and of the neonate]. PMID- 19189900 TI - Summaries for patients. Immediate versus later listing for liver transplantation for alcoholic cirrhosis. PMID- 19189901 TI - Summaries for patients. Immunochemical fecal occult blood tests. PMID- 19189902 TI - Summaries for patients. Screening for skin cancer: U.S. Preventive Services Task Force recommendation. PMID- 19189903 TI - In the clinic. Chronic kidney disease. PMID- 19189905 TI - Comparative evaluation of immunochemical fecal occult blood tests for colorectal adenoma detection. AB - BACKGROUND: Different immunochemical fecal occult blood tests (FOBTs) have been proposed for noninvasive colorectal cancer screening. Large-scale, colonoscopy based screening studies that allow evaluation of these tests for the detection of precursor lesions are scarce. OBJECTIVE: To determine and compare performance characteristics of 6 qualitative immunochemical FOBTs for identifying colorectal adenomas among adults who attended screening colonoscopy examinations. DESIGN: Prospective screening study from January 2006 to December 2007. SETTING: 20 gastroenterology practices in Germany that did screening colonoscopy. PATIENTS: 1319 participants at average risk for colorectal neoplasia who were undergoing screening colonoscopy (mean age, 63 years; 50% men). MEASUREMENTS: 6 different qualitative immunochemical FOBTs were done with stool samples collected before bowel preparation for colonoscopy. Performance characteristics (sensitivity, specificity, predictive values, and likelihood ratios) of tests were measured by comparing test results with findings on colonoscopy. Technicians who read the tests were blinded to colonoscopy results, and colonoscopists were blinded to FOBT results. RESULTS: Overall, 405 participants (31%) had an adenoma and 130 participants (10%) had an advanced adenoma. Performance characteristics varied widely among tests. For the 2 best-performing tests (immoCARE-C [CAREdiagnostica, Voerde, Germany] and FOB advanced [ulti med, Ahrensburg, Germany]), the sensitivity for detection of advanced adenomas was 25% (95% CI, 18% to 34%) and 27% (CI, 20% to 35%), respectively; specificity was 97% (CI, 95% to 98%) and 93% (CI, 91% to 95%); and the positive likelihood ratio was 3.5 (CI, 2.2 to 5.4) and 2.5 (CI, 1.9 to 3.5). LIMITATION: The study differed from real-life conditions in that stool samples were not directly dissolved in a buffer-filled vial; instead, a small container was used and stool was frozen before testing. CONCLUSION: Qualitative immunochemical FOBTs could be an option for future colorectal cancer screening because they showed better performance characteristics for precursor lesions than guaiac-based FOBTs and are practical for mass screening. However, given the large differences in diagnostic performance among tests, careful evaluation of the different test variants is important. FUNDING: The German Research Foundation (Deutsche Forschungsgemeinschaft) within the framework of a PhD program (Graduiertenkolleg 793). PMID- 19189904 TI - Immediate listing for liver transplantation versus standard care for Child-Pugh stage B alcoholic cirrhosis: a randomized trial. AB - BACKGROUND: Liver transplantation improves survival of patients with end-stage (Child-Pugh stage C) alcoholic cirrhosis, but its benefit for patients with stage B disease is uncertain. OBJECTIVE: To compare the outcomes of patients with Child Pugh stage B alcoholic cirrhosis who are immediately listed for liver transplantation with those of patients assigned to standard treatment with delay of transplantation until progression to stage C disease. DESIGN: Randomized, controlled trial. SETTING: 13 liver transplantation programs in France. PATIENTS: 120 patients with Child-Pugh stage B alcoholic cirrhosis and no viral hepatitis, cancer, or contraindication to transplantation. INTERVENTIONS: Patients were randomly assigned to immediate listing for liver transplantation (60 patients) or standard care (60 patients). MEASUREMENTS: Overall and cancer-free survival over 5 years. RESULTS: Sixty-eight percent of patients assigned to immediate listing for liver transplantation and 25% of those assigned to standard care received a liver transplant. All-cause death and cirrhosis-related death did not statistically differ between the 2 groups: 5-year survival was 58% (95% CI, 43% to 70%) for those assigned to immediate listing versus 69% (CI, 54% to 80%) for those assigned to standard care. In multivariate analysis, independent predictors of long-term survival were absence of ongoing alcohol consumption (hazard ratio, 7.604 [CI, 2.395 to 24.154]), recovery from Child-Pugh stage C (hazard ratio, 7.633 [CI, 2.392 to 24.390]), and baseline Child-Pugh score less than 8 (hazard ratio, 2.664 [CI, 1.052 to 6.746]). Immediate listing for transplantation was associated with an increased risk for extrahepatic cancer: The 5-year cancer-free survival rate was 63% (CI, 43% to 77%) for patients who were immediately listed and 94% (CI, 81% to 98%) for those who received standard care. LIMITATION: Restriction of the study sample to alcoholic patients may limit the generalizability of results to other settings. CONCLUSION: Immediate listing for liver transplantation did not show a survival benefit compared with standard care for Child-Pugh stage B alcoholic cirrhosis. In addition, immediate listing for transplantation increased the risk for extrahepatic cancer. FUNDING: The French National Program for Clinical Research. PMID- 19189906 TI - Contrast volume during primary percutaneous coronary intervention and subsequent contrast-induced nephropathy and mortality. AB - BACKGROUND: Contrast-induced nephropathy (CIN) frequently occurs in patients with acute ST-segment elevation myocardial infarction (STEMI) who are undergoing primary percutaneous coronary intervention, and CIN is associated with a more complicated clinical course and increased mortality. OBJECTIVE: To investigate the association between absolute and weight- and creatinine-adjusted contrast volume, CIN incidence, and clinical outcome in the era of mechanical reperfusion of STEMI. DESIGN: Prospective, observational study. SETTING: A university cardiology center in Milan, Italy. PATIENTS: 561 consecutive patients with STEMI who were undergoing primary percutaneous coronary intervention. MEASUREMENTS: For each patient, the maximum contrast dose was calculated, according to the formula (5 x body weight [kg])/serum creatinine, and the contrast ratio, defined as the ratio between the contrast volume administered and the maximum dose calculated, was assessed. An increase in serum creatinine of more than 25% from baseline was defined as CIN. RESULTS: 115 (20.5%) patients developed CIN. In-hospital mortality was higher among patients with CIN than those without CIN (21.4% vs. 0.9%; P < 0.001). The maximum contrast dose was exceeded in 130 (23%) patients. Patients who received more than the maximum contrast dose (contrast ratio >1) had a more complicated in-hospital clinical course and higher mortality rate (13% vs. 2.8%; P < 0.001) than did patients with a contrast ratio less than 1. Development of CIN was associated with both contrast volume and contrast ratio. LIMITATION: The association between contrast volume and outcomes was observed in a single center and could be due to comorbid conditions, disease severity, or an unknown factor. CONCLUSION: During primary percutaneous coronary intervention for STEMI, higher contrast volume is associated with higher rates of CIN and mortality; however, further study is needed to determine whether limiting contrast volume would improve patient outcome. FUNDING: Centro Cardiologico Monzino, Institute of Cardiology, University of Milan. PMID- 19189907 TI - A reengineered hospital discharge program to decrease rehospitalization: a randomized trial. AB - BACKGROUND: Emergency department visits and rehospitalization are common after hospital discharge. OBJECTIVE: To test the effects of an intervention designed to minimize hospital utilization after discharge. DESIGN: Randomized trial using block randomization of 6 and 8. Randomly arranged index cards were placed in opaque envelopes labeled consecutively with study numbers, and participants were assigned a study group by revealing the index card. SETTING: General medical service at an urban, academic, safety-net hospital. PATIENTS: 749 English speaking hospitalized adults (mean age, 49.9 years). INTERVENTION: A nurse discharge advocate worked with patients during their hospital stay to arrange follow-up appointments, confirm medication reconciliation, and conduct patient education with an individualized instruction booklet that was sent to their primary care provider. A clinical pharmacist called patients 2 to 4 days after discharge to reinforce the discharge plan and review medications. Participants and providers were not blinded to treatment assignment. MEASUREMENTS: Primary outcomes were emergency department visits and hospitalizations within 30 days of discharge. Secondary outcomes were self-reported preparedness for discharge and frequency of primary care providers' follow-up within 30 days of discharge. Research staff doing follow-up were blinded to study group assignment. RESULTS: Participants in the intervention group (n = 370) had a lower rate of hospital utilization than those receiving usual care (n = 368) (0.314 vs. 0.451 visit per person per month; incidence rate ratio, 0.695 [95% CI, 0.515 to 0.937]; P = 0.009). The intervention was most effective among participants with hospital utilization in the 6 months before index admission (P = 0.014). Adverse events were not assessed; these data were collected but are still being analyzed. LIMITATION: This was a single-center study in which not all potentially eligible patients could be enrolled, and outcome assessment sometimes relied on participant report. CONCLUSION: A package of discharge services reduced hospital utilization within 30 days of discharge. FUNDING: Agency for Healthcare Research and Quality and National Heart, Lung, and Blood Institute, National Institutes of Health. PMID- 19189908 TI - Screening for skin cancer: U.S. Preventive Services Task Force recommendation statement. AB - DESCRIPTION: Update of the 2001 U.S. Preventive Services Task Force (USPSTF) recommendation statement on screening for skin cancer. METHODS: To update its recommendation, the USPSTF reviewed evidence published since 2001 on studies on screening effectiveness, the stage of detection by screening, and the accuracy of whole-body examination by primary care clinicians and self-examination by patients. RECOMMENDATION: The USPSTF concludes that the current evidence is insufficient to assess the balance of benefits and harms of screening for skin cancer by primary care clinicians or by patient skin self-examination. (I statement). PMID- 19189909 TI - Screening for skin cancer: an update of the evidence for the U.S. Preventive Services Task Force. AB - BACKGROUND: Skin cancer is the most commonly diagnosed cancer in the United States. The majority of skin cancer is nonmelanoma cancer, either basal cell cancer or squamous cell cancer. The incidence of both melanoma and nonmelanoma skin cancer has been increasing over the past 3 decades. In 2001, the U.S. Preventive Services Task Force (USPSTF) found insufficient evidence to recommend for or against routine screening for skin cancer by using whole-body skin examination for early detection of skin cancer. PURPOSE: To update the evidence of benefits and harms of screening for skin cancer in the general population. DATA SOURCES: MEDLINE and Cochrane Library searches from 1 June 1999 to 9 August 2005 for English-language articles; recent systematic reviews; reference lists of retrieved articles; and expert suggestions. STUDY SELECTION: English-language studies were selected to answer the following key question: Does screening in asymptomatic persons with whole-body examination by a primary care clinician or by self-examination reduce morbidity and mortality from skin cancer? Randomized, controlled trials and case-control studies of screening for skin cancer were selected. One author selected English-language studies to answer the following contextual questions: Can screening with whole-body examination by primary care clinicians or by self-examination accurately detect skin cancer? Does screening with whole-body examination or by self-examination detect melanomas at an earlier stage (thinner lesions)? DATA EXTRACTION: All studies for the key question were reviewed, abstracted, and rated for quality by using predefined USPSTF criteria. DATA SYNTHESIS: No new evidence from controlled studies was found that addressed the benefit of screening for skin cancer with a whole-body examination by a physician. One article of fair quality, which reanalyzed data from a 1996 study identified for the 2001 report for the USPSTF, provides limited but insufficient evidence on the benefit of skin self-examination in the reduction of morbidity and mortality from melanoma. LIMITATIONS: Direct evidence linking skin cancer screening to improved health outcomes is lacking. Information is limited on the accuracy of screening by physicians or patients using real patients and lesions. CONCLUSION: The limited evidence prevents accurate estimation of the benefits of screening for skin cancer in the general primary care population. PMID- 19189910 TI - Update on the methods of the U.S. Preventive Services Task Force: insufficient evidence. AB - The U.S. Preventive Services Task Force (USPSTF) seeks to provide reliable and accurate evidence-based recommendations to primary care clinicians. However, clinicians indicate frustration with the lack of guidance provided by the USPSTF when the evidence is insufficient to make a recommendation. This article describes a new USPSTF plan to commission its Evidence-based Practice Centers to collect information in 4 domains pertinent to clinical decisions about prevention and to report this information routinely. The 4 domains are potential preventable burden, potential harm of the intervention, costs (both monetary and opportunity), and current practice. The process and rationale used to select these domains are presented, along with examples of how clinicians might use the information to guide clinical decision making when evidence is insufficient. PMID- 19189912 TI - Immediate listing for liver transplantation for alcoholic cirrhosis: curbing our enthusiasm. PMID- 19189911 TI - STrengthening the REporting of Genetic Association studies (STREGA): an extension of the STROBE Statement. AB - Making sense of rapidly evolving evidence on genetic associations is crucial to making genuine advances in human genomics and the eventual integration of this information into the practice of medicine and public health. Assessment of the strengths and weaknesses of this evidence, and hence the ability to synthesize it, has been limited by inadequate reporting of results. The STrengthening the REporting of Genetic Association studies (STREGA) initiative builds on the STrengthening the Reporting of Observational Studies in Epidemiology (STROBE) Statement and provides additions to 12 of the 22 items on the STROBE checklist. The additions concern population stratification, genotyping errors, modeling haplotype variation, Hardy-Weinberg equilibrium, replication, selection of participants, rationale for choice of genes and variants, treatment effects in studying quantitative traits, statistical methods, relatedness, reporting of descriptive and outcome data, and issues of data volume that are important to consider in genetic association studies. The STREGA recommendations do not prescribe or dictate how a genetic association study should be designed but seek to enhance the transparency of its reporting, regardless of choices made during design, conduct, or analysis. PMID- 19189913 TI - Exposing physicians. PMID- 19189914 TI - The unintended, negative consequences of the door-to-antibiotic measure for pneumonia. PMID- 19189915 TI - The unintended, negative consequences of the door-to-antibiotic measure for pneumonia. PMID- 19189916 TI - Are age-based criteria the best way to determine eligibility for prostate cancer screening? PMID- 19189917 TI - The unintended, negative consequences of the door-to-antibiotic measure for pneumonia. PMID- 19189918 TI - Are age-based criteria the best way to determine eligibility for prostate cancer screening? PMID- 19189919 TI - Risk prediction versus diagnosis: preserving clinical nuance in a binary world. PMID- 19189920 TI - Risk prediction versus diagnosis: preserving clinical nuance in a binary world. PMID- 19189921 TI - Risk prediction versus diagnosis: preserving clinical nuance in a binary world. PMID- 19189922 TI - Risk prediction versus diagnosis: preserving clinical nuance in a binary world. PMID- 19189923 TI - Delayed splenic rupture: myth or reality? PMID- 19189924 TI - Hypersensitivity myocarditis associated with azithromycin exposure. PMID- 19189925 TI - Acute hepatitis E virus infection in an HIV-infected person in the United States. PMID- 19189926 TI - Association between Streptococcus bovis and colon cancer. PMID- 19189928 TI - Common polymorphic transcript variation in human disease. AB - Most human genes are thought to express different transcript isoforms in different cell types; however, the full extent and functional consequences of polymorphic transcript variation (PTV), which differ between individuals within the same cell type, are unknown. Here we show that PTV is widespread in B-cells from two human populations. Tens of thousands of exons were found to be polymorphically expressed in a heritable fashion, and over 1000 of these showed strong correlations with single nucleotide polymorphism (SNP) genotypes in cis. The SNPs associated with PTV display signs of having been subject to recent positive selection in humans, and they are also highly enriched for SNPs implicated by recent genome-wide association studies of four autoimmune diseases. From this disease-association overlap, we infer that PTV is the likely mechanism by which eight common polymorphisms contribute to disease risk. A catalog of PTV will be a valuable resource for interpreting results from future disease association studies and understanding the spectrum of phenotypic differences among humans. PMID- 19189927 TI - Performance of modified universal sample processing method in a field study in Uganda. PMID- 19189929 TI - The ketogenic diet: additional information from a crossover study. AB - The purpose of this blinded, crossover study of the ketogenic diet in children with the Lennox-Gastaut syndrome was to confirm, by the addition of 60 g of glucose per day to negate the ketosis, that the effectiveness of the ketogenic diet was neither the result of a placebo effect nor due to parental expectations and commitment. We found that the additional glucose did not significantly alter the frequency of electroencephalography-assessed events, but did decrease the frequency of parent-reported "drop" seizures (P = .07). Fasting had substantial effects on both seizures and electroencephalography-assessed events. The diet remained effective in decreasing seizures of the Lennox-Gastaut syndrome at 12 days, 6 months, and 12 months. In conclusion, the ketogenic diet is effective in decreasing the drop seizures of the Lennox-Gastaut syndrome. PMID- 19189930 TI - Phenotypic variations in 3 children with POLG1 mutations. AB - Autosomal inherited mitochondrial diseases have been of increasing interest among clinicians and mitochondrial research groups because these diseases are caused through a secondary effect on the mitochondrial DNA. It was thought that the genetic stability of mitochondrial DNA relies on the accuracy of DNA polymerase gamma. Mutations of DNA polymerase gamma 1 gene (MIM# 174763) have been shown to be a cause of mitochondrial disorders associated with Mendelian disorders characterized by multiple mitochondrial DNA deletions or depletions. To date, several clinical phenotypes associated with polymerase gamma mutation have been reported presenting in both adults and children. We present 3 children in whom were found to have reported pathogenic DNA polymerase gamma 1 mutations: heterozygous p.G517V in 2 half siblings and heterozygous p.T251I and p.P587L in the other. The aim of this communication is to report 3 pediatric cases associated with DNA polymerase gamma 1 mutations to augment the expanding clinical phenotype that has been previously reported. PMID- 19189931 TI - Methyl-CpG-binding protein 2 (MECP2) gene mutations in an Italian sample of patients with pervasive developmental disorder and mental retardation. AB - Methyl-CpG-binding protein 2 (MECP2) gene mutations have been identified in girls with Rett syndrome and in boys with heterogeneous neuropsychiatric disorders. Because of the limited or inconsistent data reported in literature, the role of methyl-CpG-binding protein 2 gene in the pathogenesis of mental retardation and pervasive developmental disorders needs further study. We scanned methyl-CpG binding protein 2 gene in 99 Italian patients with pervasive developmental disorder or with nonsyndromal mental retardation. Four methyl-CpG-binding protein 2 gene mutations were found: 2 in 4 girls with Rett disorder, the others in 2 girls with mental retardation. The wide phenotypic spectrum and the variants of methyl-CpG-binding protein 2 gene, which may play an important role in gene regulation and neurodevelopment, justify the literature's interest particularly in girls. PMID- 19189932 TI - Acute tumefactive demyelinating lesions in a pediatric patient with known diagnosis of multiple sclerosis: review of the literature and treatment proposal. AB - Tumefactive demyelinating lesions are a known but uncommon complication of multiple sclerosis, a disease rarely reported in children. This is the case of a 16-year-old African American patient with multiple sclerosis, who developed 2 tumefactive demyelinating lesions. Review of the literature and our own experience helped formulate an algorithm for therapeutic options during an acute attack. PMID- 19189933 TI - Kindergarten children's failure to qualify for first grade could result from sleep disturbances. AB - Every year, 7% to 15% of preschool children are found to be underqualified for first grade. We examined whether sleep disturbances are factors in school readiness and their association with neurocognitive skills and behavior. The population included 148 kindergarten students. The study group consisted of 50 students who were assessed by the educational authority as unready for first grade. Children who were scheduled to attend first grade (n = 98) were in the control group. All children/parents filled in a sleep questionnaire and underwent a week of actigraphic sleep/wake study as well as cognitive and behavioral assessments. Children in the study group had significantly shorter total sleep time, reduced sleep efficiency, and increased number of nighttime awakenings. There were significant correlations between sleep variables, and cognitive and behavioral scores. In conclusion, children who fail to qualify for first grade have significantly inferior sleep patterns. Sleep disturbances were associated with cognitive and emotional immaturity. PMID- 19189934 TI - The underrecognized epilepsy spectrum: the effects of levetiracetam on neuropsychological functioning in relation to subclinical spike production. AB - The purpose of this prospective, open-label pilot study was to determine whether treatment with levetiracetam improves neuropsychological functioning in children and adolescents who have evidence of subclinical spike production associated with attention and learning difficulties. Six participants (mean age 9.8 years) were treated with levetiracetam up to 40 mg/kg per day and evaluated using neuropsychological (Wide Range Assessment of Memory and Learning, Second Edition), academic (Wechsler Individual Achievement Test, Second Edition, Abbreviated), and electroencephalographic assessments at baseline and after 10 weeks of treatment. Statistically significant improvements on indexes of the Wide Range Assessment of Memory and Learning, Second Edition were observed in 4 participants after 10 weeks. No statistically significant differences were observed for the Wechsler Individual Achievement Test, Second Edition, Abbreviated. Concomitant spike suppression was observed. Levetiracetam was generally well tolerated. A subset of patients exists with attention and learning problems that have associated aberrant cortical electrical activity without clinical seizures and associated neuropsychological deficits that may improve after treatment with levetiracetam. PMID- 19189935 TI - Co-occurrence of intracerebral tuberculoma with lumbar intramedullary tuberculoma. AB - Tuberculosis is common in India, but the co-occurrence of intracranial and intramedullary tuberculomas as a manifestation of central nervous system tuberculosis is extremely rare in children. We report a case of concurrent intracranial and lumbar intramedullary tuberculomas in a 6-year-old girl. The child developed the intracranial and intramedullary tuberculomas while on antituberculous therapy for previously diagnosed tuberculous meningitis. The child improved well on antituberculous drugs and neurosurgical excision of the lumbar lesion. Histopathology confirmed tuberculous etiology of the intramedullary lesion. PMID- 19189936 TI - Essential tremor after ipsilateral cerebellar hemispherectomy: support for the thalamus as the central oscillator. AB - Essential tremor is a long-recognized and common movement disorder, yet controversy still surrounds its pathophysiology. The olivo-cerebello-thalamo cortical pathway has been implicated in the genesis of essential tremor, and the inferior olive has been considered the central oscillator driving the peripheral tremor. We present the case of a patient who developed essential tremor ipsilateral to cerebellar hemispherectomy and propose that the central oscillator in patients with essential tremor may not be the inferior olive in all cases, but rather the nucleus ventralis intermedius of the thalamus. PMID- 19189937 TI - TNF-alpha stimulates the ACAT1 expression in differentiating monocytes to promote the CE-laden cell formation. AB - High levels of the inflammatory cytokine tumor necrosis factor-alpha (TNF-alpha) are present in atherosclerotic lesions. TNF-alpha regulates expression of multiple genes involved in various stages of atherosclerosis, and it exhibits proatherosclerotic and antiatherosclerotic properties. ACAT catalyzes the formation of cholesteryl esters (CE) in monocytes/macrophages, and it promotes the foam cell formation at the early stage of atherosclerosis. We hypothesize that TNF-alpha may be involved in regulating the ACAT gene expression in monocytes/macrophages. In this article, we show that in cultured, differentiating human monocytes, TNF-alpha enhances the expression of the ACAT1 but not ACAT2 gene, increases the cholesteryl ester accumulation, and promotes the lipid-laden cell formation. Several other proinflammatory cytokines tested do not affect the ACAT1 gene expression. The stimulation effect is consistent with a receptor dependent process, and is blocked by using nuclear factor-kappa B (NF-kappa B) inhibitors. A functional and unique NF-kappa B element located within the human ACAT1 gene proximal promoter is required to mediate the action of TNF-alpha. Our data demonstrate that TNF-alpha, through the NF-kappa B pathway, specifically enhances the expression of human ACAT1 gene to promote the CE-laden cell formation from the differentiating monocytes, and our data support the hypothesis that TNF-alpha is proatherosclerotic during early phase of lesion development. PMID- 19189940 TI - Motor training of upper extremity with functional electrical stimulation in early stroke rehabilitation. AB - BACKGROUND: Functional electrical stimulation (FES) allows active exercises in stroke patients with upper extremity paralysis. OBJECTIVE: To investigate the effect of motor training with FES on motor recovery in acute and subacute stroke patients with severe to complete arm and/or hand paralysis. METHODS: For this pilot study, 23 acute and subacute stroke patients were randomly assigned to the intervention (n = 12) and control group (n = 11). Distributed over 4 weeks, FES training replaced 12 conventional training sessions in the intervention group. An Extended Barthel Index (EBI) subscore assessed the performance of activities of daily living (ADL). The Chedoke McMaster Stroke Assessment (CMSA) measured hand and arm function and shoulder pain. The Modified Ashworth Scale (MAS) assessed resistance to passive movement. Unblinded assessments were performed prior to and following the end of the training period. RESULTS: The EBI subscore and CMSA arm score improved significantly in both groups. The CMSA hand function improved significantly in the FES group. Resistance to passive movement of finger and wrist flexors increased significantly in the FES group. Shoulder pain did not change significantly. None of the outcome measures, however, demonstrated significant gain differences between the groups. CONCLUSIONS: We did not find clear evidence for superiority or inferiority of FES. Our findings, and those of similar trials, suggest that the number of sessions should be at least doubled to test for superiority of FES in these highly impaired patients and approximately 50 participants would have to be assigned to each therapeutic intervention to find significant differences. PMID- 19189939 TI - The future of restorative neurosciences in stroke: driving the translational research pipeline from basic science to rehabilitation of people after stroke. AB - BACKGROUND: Major advances during the past 50 years highlight the immense potential for restoration of function after neural injury, even in the damaged adult human brain. Yet, the translation of these advances into clinically useful treatments is painstakingly slow. OBJECTIVE: Here, we consider why the traditional model of a "translational research pipeline" that transforms basic science into novel clinical practice has failed to improve rehabilitation practice for people after stroke. RESULTS: We find that (1) most treatments trialed in vitro and in animal models have not yet resulted in obviously useful functional gains in patients; (2) most clinical trials of restorative treatments after stroke have been limited to small-scale studies; (3) patient recruitment for larger clinical trials is difficult; (4) the determinants of patient outcomes and what patients want remain complex and ill-defined, so that basic scientists have no clear view of the clinical importance of the problems that they are addressing; (5) research in academic neuroscience centers is poorly integrated with practice in front-line hospitals and the community, where the majority of patients are treated; and (6) partnership with both industry stakeholders and patient pressure groups is poorly developed, at least in the United Kingdom where research in the translational restorative neurosciences in stroke depends on public sector research funds and private charities. CONCLUSIONS: We argue that interaction between patients, front-line clinicians, and clinical and basic scientists is essential so that they can explore their different priorities, skills, and concerns. These interactions can be facilitated by funding research consortia that include basic and clinical scientists, clinicians and patient/carer representatives with funds targeted at those impairments that are major determinants of patient and carer outcomes. Consortia would be instrumental in developing a lexicon of common methods, standardized outcome measures, data sharing and long-term goals. Interactions of this sort would create a research friendly, rather than only target-led, culture in front-line stroke rehabilitation services. PMID- 19189942 TI - Suppression of the double-strand-break-repair defect of the Saccharomyces cerevisiae rad57 mutant. AB - The Rad51 paralogs Rad55 and Rad57 form a heterodimer required to mediate the formation and/or stabilization of the Rad51 filament. To further characterize the function of Rad55-Rad57, we used a combination of rad57 partial suppressors to determine whether the DNA repair and recombination defects of the rad57 mutant could be completely suppressed. The combination of all suppressors, elevated temperature, srs2, rad51-I345T, and mating-type (MAT) heterozygosity resulted in almost complete suppression of the rad57 mutant defect in the recruitment of Rad51 to DNA-damaged sites, as well as survival in response to ionizing radiation and camptothecin. In a physical assay to monitor the kinetics of double-strand break (DSB)-induced gene conversion, the rad57 mutant defect was effectively suppressed by srs2 and MAT heterozygosity, but these same suppressors failed to suppress the spontaneous recombination defect. Thus the Rad55-Rad57 heterodimer appears to have a unique function in spontaneous recombination that is not essential for DSB repair. Furthermore, we investigated the currently unknown mechanism of rad57 suppression by MAT heterozygosity and found that it is independent of DNL4. PMID- 19189944 TI - Multiple SET methyltransferases are required to maintain normal heterochromatin domains in the genome of Drosophila melanogaster. AB - Methylation of histone H3 lysine 9 (H3K9) is a key feature of silent chromatin and plays an important role in stabilizing the interaction of heterochromatin protein 1 (HP1) with chromatin. Genomes of metazoans such as the fruit fly Drosophila melanogaster generally encode three types of H3K9-specific SET domain methyltransferases that contribute to chromatin homeostasis during the life cycle of the organism. SU(VAR)3-9, dG9a, and dSETDB1 all function in the generation of wild-type H3K9 methylation levels in the Drosophila genome. Two of these enzymes, dSETDB1 and SU(VAR)3-9, govern heterochromatin formation in distinct but overlapping patterns across the genome. H3K9 methylation in the small, heterochromatic fourth chromosome of D. melanogaster is governed mainly by dSETDB1, whereas dSETDB1 and SU(VAR)3-9 function in concert to methylate H3K9 in the pericentric heterochromatin of all chromosomes, with dG9a having little impact in these domains, as shown by monitoring position effect variegation. To understand how these distinct heterochromatin compartments may be differentiated, we examined the developmental timing of dSETDB1 function using a knockdown strategy. dSETDB1 acts to maintain heterochromatin during metamorphosis, at a later stage in development than the reported action of SU(VAR)3-9. Surprisingly, depletion of both of these enzymes has less deleterious effect than depletion of one. These results imply that dSETDB1 acts as a heterochromatin maintenance factor that may be required for the persistence of earlier developmental events normally governed by SU(VAR)3-9. In addition, the genetic interactions between dSETDB1 and Su(var)3-9 mutations emphasize the importance of maintaining the activities of these histone methyltransferases in balance for normal genome function. PMID- 19189945 TI - De Novo identification of single nucleotide mutations in Caenorhabditis elegans using array comparative genomic hybridization. AB - Array comparative genomic hybridization (aCGH) has been used primarily to detect copy-number variants between two genomes. Here we report using aCGH to detect single nucleotide mutations on oligonucleotide microarrays with overlapping 50 mer probes. This technique represents a powerful method for rapidly detecting novel homozygous single nucleotide mutations in any organism with a sequenced reference genome. PMID- 19189946 TI - Episodic selection and the maintenance of competence and natural transformation in Bacillus subtilis. AB - We present a new hypothesis for the selective pressures responsible for maintaining natural competence and transformation. Our hypothesis is based in part on the observation that in Bacillus subtilis, where transformation is widespread, competence is associated with periods of nongrowth in otherwise growing populations. As postulated for the phenomenon of persistence, the short term fitness cost associated with the production of transiently nongrowing bacteria can be compensated for and the capacity to produce these competent cells can be favored due to episodes where the population encounters conditions that kill dividing bacteria. With the aid of a mathematical model, we demonstrate that under realistic conditions this "episodic selection" for transiently nongrowing (persisting) bacteria can maintain competence for the uptake and expression of exogenous DNA transformation. We also show that these conditions for maintaining competence are dramatically augmented even by rare episodes where selection favors transformants. Using experimental populations of B. subtilis and antibiotic-mediated episodic selection, we test and provide support for the validity of the assumptions behind this model and the predictions generated from our analysis of its properties. We discuss the potential generality of episodic selection for the maintenance of competence in other naturally transforming species of bacteria and critically evaluate other hypotheses for the maintenance (and evolution) of competence and their relationship to this hypothesis. PMID- 19189947 TI - GPC-1, a G protein gamma-subunit, regulates olfactory adaptation in Caenorhabditis elegans. AB - Caenorhabditis elegans genome carries two Ggamma genes, gpc-1 and gpc-2, and two Gbeta genes, gpb-1 and gpb-2. Of these, gpc-2 and gpb-1 are expressed ubiquitously and are essential for viability. Through a genetic screen, we identified gpc-1 as essential for olfactory adaptation. While wild-type animals show decreased chemotaxis to the odorant benzaldehyde after a short preexposure to the odorant, gpc-1 mutants are still attracted to the odorant after the same preexposure. Cell-specific rescue experiments show that gpc-1 acts in the AWC olfactory neurons. Coexpression of GPC-1 and GPB-1, but not GPB-2, caused enhanced adaptation, indicating that GPC-1 may act with GPB-1. On the other hand, knock down of gpc-2 by cell-targeted RNAi caused reduced chemotaxis to the odorant in unadapted animals, indicating that GPC-2 mainly act for olfactory sensation and the two Ggamma's have differential functions. Nonetheless, overexpression of gpc-2 in AWC neurons rescued the adaptation defects of gpc-1 mutants, suggesting partially overlapping functions of the two Ggamma's. We further tested genetic interaction of gpc-1 with several other genes involved in olfactory adaptation. Our analyses place goa-1 Goalpha and let-60 Ras in parallel to gpc-1. In contrast, a gain-of-function mutation in egl-30 Gqalpha was epistatic to gpc-1, suggesting the possibility that gpc-1 Ggamma may act upstream of egl-30 Gqalpha. PMID- 19189948 TI - Waiting longer for two mutations. PMID- 19189949 TI - Linkage disequilibrium and demographic history of wild and domestic canids. AB - Assessing the extent of linkage disequilibrium (LD) in natural populations of a nonmodel species has been difficult due to the lack of available genomic markers. However, with advances in genotyping and genome sequencing, genomic characterization of natural populations has become feasible. Using sequence data and SNP genotypes, we measured LD and modeled the demographic history of wild canid populations and domestic dog breeds. In 11 gray wolf populations and one coyote population, we find that the extent of LD as measured by the distance at which r2=0.2 extends <10 kb in outbred populations to >1.7 Mb in populations that have experienced significant founder events and bottlenecks. This large range in the extent of LD parallels that observed in 18 dog breeds where the r2 value varies from approximately 20 kb to >5 Mb. Furthermore, in modeling demographic history under a composite-likelihood framework, we find that two of five wild canid populations exhibit evidence of a historical population contraction. Five domestic dog breeds display evidence for a minor population contraction during domestication and a more severe contraction during breed formation. Only a 5% reduction in nucleotide diversity was observed as a result of domestication, whereas the loss of nucleotide diversity with breed formation averaged 35%. PMID- 19189950 TI - Identification of receptor-tyrosine-kinase-signaling target genes reveals receptor-specific activities and pathway branchpoints during Drosophila development. AB - Receptor tyrosine kinases (RTKs) are an important family of signaling molecules with the unusual property that they are able to transduce their signals using the same downstream pathways. This has led to an unresolved debate as to whether individual receptors are interchangeable, or if each receptor can mediate specific downstream responses. To address this question, we have conducted a screen to identify target genes whose expression is differentially modulated by RTKs and their downstream pathway components. Using whole-mount in situ hybridization in Drosophila embryos exposed to constitutively active RTK pathway signaling, along with quantitative RT-PCR, we found that a significant fraction of target genes respond differentially in a spatial and/or quantitative manner. This includes differential responses to EGF receptor vs. fibroblast growth factor receptor signaling as well as to more downstream components such as Ras1 and pointed. We show that not only genes but also individual alternative transcripts can respond differently to signaling, and we present evidence that the differential responses can be mediated at the transcriptional level. Our results demonstrate that different RTKs can elicit distinct transcriptional responses, and the target genes obtained from our screen provide a valuable resource for further exploration of the mechanisms underlying this signaling specificity. PMID- 19189951 TI - Genetics and lineage-specific evolution of a lethal hybrid incompatibility between Drosophila mauritiana and its sibling species. AB - The Dobzhansky-Muller model posits that intrinsic postzygotic reproductive isolation--the sterility or lethality of species hybrids--results from the evolution of incompatible epistatic interactions between species: favorable or neutral alleles that become fixed in the genetic background of one species can cause sterility or lethality in the genetic background of another species. The kind of hybrid incompatibility that evolves between two species, however, depends on the particular evolutionary history of the causative substitutions. An allele that is functionally derived in one species can be incompatible with an allele that is functionally derived in the other species (a derived-derived hybrid incompatibility). But an allele that is functionally derived in one species can also be incompatible with an allele that has retained the ancestral state in the other species (a derived-ancestral hybrid incompatibility). The relative abundance of such derived-derived vs. derived-ancestral hybrid incompatibilities is unknown. Here, we characterize the genetics and evolutionary history of a lethal hybrid incompatibility between Drosophila mauritiana and its two sibling species, D. sechellia and D. simulans. We show that a hybrid lethality factor(s) in the pericentric heterochromatin of the D. mauritiana X chromosome, hybrid lethal on the X (hlx), is incompatible with a factor(s) in the same small autosomal region from both D. sechellia and D. simulans, Suppressor of hlx [Su(hlx)]. By combining genetic and phylogenetic information, we infer that hlx Su(hlx) hybrid lethality is likely caused by a derived-ancestral incompatibility, a hypothesis that can be tested directly when the genes are identified. PMID- 19189952 TI - Approximate bayesian computation without summary statistics: the case of admixture. AB - In recent years approximate Bayesian computation (ABC) methods have become popular in population genetics as an alternative to full-likelihood methods to make inferences under complex demographic models. Most ABC methods rely on the choice of a set of summary statistics to extract information from the data. In this article we tested the use of the full allelic distribution directly in an ABC framework. Although the ABC techniques are becoming more widely used, there is still uncertainty over how they perform in comparison with full-likelihood methods. We thus conducted a simulation study and provide a detailed examination of ABC in comparison with full likelihood in the case of a model of admixture. This model assumes that two parental populations mixed at a certain time in the past, creating a hybrid population, and that the three populations then evolve under pure drift. Several aspects of ABC methodology were investigated, such as the effect of the distance metric chosen to measure the similarity between simulated and observed data sets. Results show that in general ABC provides good approximations to the posterior distributions obtained with the full-likelihood method. This suggests that it is possible to apply ABC using allele frequencies to make inferences in cases where it is difficult to select a set of suitable summary statistics and when the complexity of the model or the size of the data set makes it computationally prohibitive to use full-likelihood methods. PMID- 19189953 TI - Recombination rates may affect the ratio of X to autosomal noncoding polymorphism in African populations of Drosophila melanogaster. PMID- 19189954 TI - Cis-regulatory mutations in the Caenorhabditis elegans homeobox gene locus cog-1 affect neuronal development. AB - We apply here comparative genome hybridization as a novel tool to identify the molecular lesion in two Caenorhabditis elegans mutant strains that affect a neuronal cell fate decision. The phenotype of the mutant strains resembles those of the loss-of-function alleles of the cog-1 homeobox gene, an inducer of the fate of the gustatory neuron ASER. We find that both lesions map to the cis regulatory control region of cog-1 and affect a phylogenetically conserved binding site for the C2H2 zinc-finger transcription factor CHE-1, a previously known regulator of cog-1 expression in ASER. Identification of this CHE-1-binding site as a critical regulator of cog-1 expression in the ASER in vivo represents one of the rare demonstrations of the in vivo relevance of an experimentally determined or predicted transcription-factor-binding site. Aside from the mutationally defined CHE-1-binding site, cog-1 contains a second, functional CHE 1-binding site, which in isolation is sufficient to drive reporter gene expression in the ASER but in an in vivo context is apparently insufficient for promoting appropriate ASER expression. The cis-regulatory control regions of other ASE-expressed genes also contain ASE motifs that can promote ASE neuron expression when isolated from their genomic context, but appear to depend on multiple ASE motifs in their normal genomic context. The multiplicity of cis regulatory elements may ensure the robustness of gene expression. PMID- 19189955 TI - A domestic cat X chromosome linkage map and the sex-linked orange locus: mapping of orange, multiple origins and epistasis over nonagouti. AB - A comprehensive genetic linkage map of the domestic cat X chromosome was generated with the goal of localizing the genomic position of the classic X linked orange (O) locus. Microsatellite markers with an average spacing of 3 Mb were selected from sequence traces of the cat 1.9x whole genome sequence (WGS), including the pseudoautosomal region 1 (PAR1). Extreme variation in recombination rates (centimorgans per megabase) was observed along the X chromosome, ranging from a virtual absence of recombination events in a region estimated to be >30 Mb to recombination frequencies of 15.7 cM/Mb in a segment estimated to be <0.3 Mb. This detailed linkage map was applied to position the X-linked orange gene, placing this locus on the q arm of the X chromosome, as opposed to a previously reported location on the p arm. Fine mapping placed the locus between markers at positions 106 and 116.8 Mb in the current 1.9x-coverage sequence assembly of the cat genome. Haplotype analysis revealed potential recombination events that could reduce the size of the candidate region to 3.5 Mb and suggested multiple origins for the orange phenotype in the domestic cat. Furthermore, epistasis of orange over nonagouti was demonstrated at the genetic level. PMID- 19189956 TI - The decay of the chromosomally encoded ccdO157 toxin-antitoxin system in the Escherichia coli species. AB - The origin and the evolution of toxin-antitoxin (TA) systems remain to be uncovered. TA systems are abundant in bacterial chromosomes and are thought to be part of the flexible genome that originates from horizontal gene transfer. To gain insight into TA system evolution, we analyzed the distribution of the chromosomally encoded ccdO157 system in 395 natural isolates of Escherichia coli. It was discovered in the E. coli O157:H7 strain in which it constitutes a genomic islet between two core genes (folA and apaH). Our study revealed that the folA apaH intergenic region is plastic and subject to insertion of foreign DNA. It could be composed (i) of a repetitive extragenic palindromic (REP) sequence, (ii) of the ccdO157 system or subtle variants of it, (iii) of a large DNA piece that contained a ccdAO157 antitoxin remnant in association with ORFs of unknown function, or (iv) of a variant of it containing an insertion sequence in the ccdAO157 remnant. Sequence analysis and functional tests of the ccdO157 variants revealed that 69% of the variants were composed of an active toxin and antitoxin, 29% were composed of an active antitoxin and an inactive toxin, and in 2% of the cases both ORFs were inactive. Molecular evolution analysis showed that ccdBO157 is under neutral evolution, suggesting that this system is devoid of any biological role in the E. coli species. PMID- 19189957 TI - Genetics of Sex-linked yellow in the Syrian hamster. AB - Alternating patches of black and yellow pigment are a ubiquitous feature of mammalian color variation that contributes to camouflage, species recognition, and morphologic diversity. X-linked determinants of this pattern--recognized by variegation in females but not in males--have been described in the domestic cat as Orange, and in the Syrian hamster as Sex-linked yellow (Sly), but are curiously absent from other vertebrate species. Using a comparative genomic approach, we develop molecular markers and a linkage map for the euchromatic region of the Syrian hamster X chromosome that places Sly in a region homologous to the centromere-proximal region of human Xp. Comparison to analogous work carried out for Orange in domestic cats indicates, surprisingly, that the cat and hamster mutations lie in nonhomologous regions of the X chromosome. We also identify the molecular cause of recessively inherited black coat color in hamsters (historically referred to as nonagouti) as a Cys115Tyr mutation in the Agouti gene. Animals doubly mutant for Sly and nonagouti exhibit a Sly phenotype. Our results indicate that Sly represents a melanocortin pathway component that acts similarly to, but is genetically distinct from, Mc1r and that has implications for understanding both the evolutionary history and the mutational mechanisms of pigment-type switching. PMID- 19189958 TI - Fission yeast rgf2p is a rho1p guanine nucleotide exchange factor required for spore wall maturation and for the maintenance of cell integrity in the absence of rgf1p. AB - Schizosaccharomyces pombe Rho1p is essential, directly activates beta-1,3-glucan synthase, and participates in the regulation of morphogenesis. In S. pombe, Rho1p is activated by at least three guanine nucleotide exchange factors (GEFs): Rgf1p, Rgf2p, and Rgf3p. In this study we show that Rgf2p is a Rho1p GEF required for sporulation. The rgf2+ deletion did not affect forespore membrane formation and the nuclei were encapsulated properly. However, the mutant ascospores appeared dark and immature. The rgf2Delta zygotes were not able to release the ascospores spontaneously, and the germination efficiency was greatly reduced compared to wild-type (wt) spores. This phenotype resembles that of the mutants in bgs2+, which encodes a sporulation-specific glucan synthase subunit. In fact, glucan synthase activity was diminished in sporulating rgf2Delta diploids. Rgf2p also plays a role in beta-glucan biosynthesis during vegetative growth. Overexpression of rgf2+ specifically increased GTP-bound Rho1p, caused changes in cell morphology, and elicited an increase in beta-1,3-glucan synthase activity. Moreover, the simultaneous disruption of rgf1+ and rgf2+ was lethal and both Rgf1p and Rgf2p were able to partially substitute for each other. Our results suggest that Rgf1p and Rgf2p are alternative GEFs with an essential overlapping function in Rho1p activation during vegetative growth. PMID- 19189960 TI - Dextromethorphan reduces oxidative stress and inhibits atherosclerosis and neointima formation in mice. AB - AIMS: Macrophage-related oxidative stress plays an important role in the inflammatory process in atherosclerosis. Recently, dextromethorphan (DXM), a common cough-suppressing ingredient with a high safety profile, was found to inhibit the activation of microglia, the resident macrophage in the nervous system. We investigated whether DXM could reduce macrophage production of cytokines and superoxide and the resultant influence on atherosclerosis formation in mice. METHODS AND RESULTS: We used in vitro and in vivo studies to evaluate the DXM inhibitory effect on oxidative stress. Dextromethorphan pretreatment significantly suppressed the production of tumour necrosis factor-alpha, monocyte chemoattractant protein-1, interleukin-6, interleukin-10, and superoxide in macrophage cell culture after stimulation. Indeed, DXM reduced macrophage nicotinamide adenine dinucleotide phosphate oxidase activity by decreasing membrane translocation of p47(phox) and p67(phox) through the inhibition of protein kinase C and extracellular signal-regulated kinase activation. The anti atherosclerosis effect of DXM was tested using two animal models, apolipoprotein E (apoE)-deficient mice and a mouse carotid ligation model. Dextromethorphan treatment (10-40 mg/kg/day) for 10 weeks in apoE-deficient mice significantly reduced superoxide production in their polymorphonuclear leukocytes and aortas. It significantly decreased the severity of aortic atherosclerosis in the apoE deficient mice and decreased carotid neointima formation after ligation in C57BL/6 mice. CONCLUSION: Our data show that DXM, with its novel effect in reducing oxidative stress, significantly reduces atherosclerosis and neointima formation in mice. PMID- 19189961 TI - Scaffold-forming and Adhesive Contributions of Synthetic Laminin-binding Proteins to Basement Membrane Assembly. AB - Laminins that possess three short arms contribute to basement membrane assembly by anchoring to cell surfaces, polymerizing, and binding to nidogen and collagen IV. Although laminins containing the alpha4 and alpha5 subunits are expressed in alpha2-deficient congenital muscular dystrophy, they may be ineffective substitutes because they bind weakly to cell surfaces and/or because they lack the third arm needed for polymerization. We asked whether linker proteins engineered to bind to deficient laminins that provide such missing activities would promote basement membrane assembly in a Schwann cell model. A chimeric fusion protein (alphaLNNd) that adds a short arm terminus to laminin through the nidogen binding locus was generated and compared with the dystrophy-ameliorating protein miniagrin (mAgrin) that binds to the laminin coiled-coil dystroglycan and sulfatides. alphaLNNd was found to mediate laminin binding to collagen IV, to bind to galactosyl sulfatide, and to selectively convert alpha-short arm deletion mutant laminins LmDeltaalphaLN and LmDeltaalphaLN-L4b into polymerizing laminins. This protein enabled polymerization-deficient laminin but not an adhesion deficient laminin lacking LG domains (LmDeltaLG) to assemble an extracellular matrix on Schwann cell surfaces. mAgrin, on the other hand, enabled LmDeltaLG to form an extracellular matrix on cell surfaces without increasing accumulation of non-polymerizing laminins. These gain-of-function studies reveal distinct polymerization and anchorage contributions to basement membrane assembly in which the three different LN domains mediate the former, and the LG domains provide primary anchorage with secondary contributions from the alphaLN domain. These findings may be relevant for an understanding of the pathogenesis and treatment of laminin deficiency states. PMID- 19189962 TI - Structure-Function, Stability, and Chemical Modification of the Cyanobacterial Cytochrome b6f Complex from Nostoc sp. PCC 7120. AB - The crystal structure of the cyanobacterial cytochrome b(6)f complex has previously been solved to 3.0-A resolution using the thermophilic Mastigocladus laminosus whose genome has not been sequenced. Several unicellular cyanobacteria, whose genomes have been sequenced and are tractable for mutagenesis, do not yield b(6)f complex in an intact dimeric state with significant electron transport activity. The genome of Nostoc sp. PCC 7120 has been sequenced and is closer phylogenetically to M. laminosus than are unicellular cyanobacteria. The amino acid sequences of the large core subunits and four small peripheral subunits of Nostoc are 88 and 80% identical to those in the M. laminosus b(6)f complex. Purified b(6)f complex from Nostoc has a stable dimeric structure, eight subunits with masses similar to those of M. laminosus, and comparable electron transport activity. The crystal structure of the native b(6)f complex, determined to a resolution of 3.0A (PDB id: 2ZT9), is almost identical to that of M. laminosus. Two unique aspects of the Nostoc complex are: (i) a dominant conformation of heme b(p) that is rotated 180 degrees about the alpha- and gamma-meso carbon axis relative to the orientation in the M. laminosus complex and (ii) acetylation of the Rieske iron-sulfur protein (PetC) at the N terminus, a post-translational modification unprecedented in cyanobacterial membrane and electron transport proteins, and in polypeptides of cytochrome bc complexes from any source. The high spin electronic character of the unique heme c(n) is similar to that previously found in the b(6)f complex from other sources. PMID- 19189959 TI - Genomewide patterns of substitution in adaptively evolving populations of the RNA bacteriophage MS2. AB - Experimental evolution of bacteriophage provides a powerful means of studying the genetics of adaptation, as every substitution contributing to adaptation can be identified and characterized. Here, I use experimental evolution of MS2, an RNA bacteriophage, to study its adaptive response to a novel environment. To this end, three lines of MS2 were adapted to rapid growth and lysis at cold temperature for a minimum of 50 phage generations and subjected to whole-genome sequencing. Using this system, I identified adaptive substitutions, monitored changes in frequency of adaptive mutations through the course of the experiment, and measured the effect on phage growth rate of each substitution. All three lines showed a substantial increase in fitness (a two- to threefold increase in growth rate) due to a modest number of substitutions (three to four). The data show some evidence that the substitutions occurring early in the experiment have larger beneficial effects than later ones, in accordance with the expected diminishing returns relationship between the fitness effects of a mutation and its order of substitution. Patterns of molecular evolution seen here--primarily a paucity of hitchhiking mutations--suggest an abundant supply of beneficial mutations in this system. Nevertheless, some beneficial mutations appear to have been lost, possibly due to accumulation of beneficial mutations on other genetic backgrounds, clonal interference, and negatively epistatic interactions with other beneficial mutations. PMID- 19189964 TI - The CoxD protein of Oligotropha carboxidovorans is a predicted AAA+ ATPase chaperone involved in the biogenesis of the CO dehydrogenase [CuSMoO2] cluster. AB - CO dehydrogenase from the Gram-negative chemolithoautotrophic eubacterium Oligotropha carboxidovorans OM5 is a structurally characterized molybdenum containing iron-sulfur flavoenzyme, which catalyzes the oxidation of CO (CO + H(2)O --> CO(2) + 2e(-) + 2H(+)). It accommodates in its active site a unique bimetallic [CuSMoO(2)] cluster, which is subject to post-translational maturation. Insertional mutagenesis of coxD has established its requirement for the assembly of the [CuSMoO(2)] cluster. Disruption of coxD led to a phenotype of the corresponding mutant OM5 D::km with the following characteristics: (i) It was impaired in the utilization of CO, whereas the utilization of H(2) plus CO(2) was not affected; (ii) Under appropriate induction conditions bacteria synthesized a fully assembled apo-CO dehydrogenase, which could not oxidize CO; (iii) Apo-CO dehydrogenase contained a [MoO(3)] site in place of the [CuSMoO(2)] cluster; and (iv) Employing sodium sulfide first and then the Cu(I)-(thiourea)(3) complex, the non-catalytic [MoO(3)] site could be reconstituted in vitro to a [CuSMoO(2)] cluster capable of oxidizing CO. Sequence information suggests that CoxD is a MoxR-like AAA+ ATPase chaperone related to the hexameric, ring-shaped BchI component of Mg(2+)-chelatases. Recombinant CoxD, which appeared in Escherichia coli in inclusion bodies, occurs exclusively in cytoplasmic membranes of O. carboxidovorans grown in the presence of CO, and its occurrence coincided with GTPase activity upon sucrose density gradient centrifugation of cell extracts. The presumed function of CoxD is the partial unfolding of apo-CO dehydrogenase to assist in the stepwise introduction of sulfur and copper in the [MoO(3)] center of the enzyme. PMID- 19189963 TI - Arabidopsis GH3.12 (PBS3) conjugates amino acids to 4-substituted benzoates and is inhibited by salicylate. AB - Salicylate (SA, 2-hydroxybenzoate) is a phytohormone best known for its role as a critical mediator of local and systemic plant defense responses. In response to pathogens such as Pseudomonas syringae, SA is synthesized and activates widespread gene expression. In gh3.12/pbs3 mutants of Arabidopsis thaliana, induced total SA accumulation is significantly compromised as is SA-dependent gene expression and plant defense. AtGH3 subfamily I and II members have been shown to conjugate phytohormone acyl substrates to amino acids in vitro, with this role supported by in planta analyses. Here we sought to determine the in vitro biochemical activity and kinetic properties of GH3.12/avrPphB susceptible 3 (PBS3), a member of the uncharacterized AtGH3 subfamily III. Using a novel high throughput adenylation assay, we characterized the acyl substrate preference of PBS3. We found PBS3 favors 4-substituted benzoates such as 4-aminobenzoate and 4 hydroxybenzoate, with moderate activity on benzoate and no observed activity with 2-substituted benzoates. Similar to known GH3 enzymes, PBS3 catalyzes the conjugation of specific amino acids (e.g. Glu) to its preferred acyl substrates. Kinetic analyses indicate 4-aminobenzoate and 4-hydroxybenzoate are preferred acyl substrates as PBS3 exhibits both higher affinities (apparent K(m) = 153 and 459 microm, respectively) and higher catalytic efficiencies (k(cat)/K(m) = 0.0179 and 0.0444 microm(-1) min(-1), respectively) with these acyl substrates compared with benzoate (apparent K(m) = 867 microm, k(cat)/K(m) = 0.0046 microm(-1) min( 1)). Notably, SA specifically and reversibly inhibits PBS3 activity with an IC(50) of 15 microm. This suggests a general mechanism for the rapid, reversible regulation of GH3 activity and small molecule cross-talk. For PBS3, this may allow for coordination of flux through diverse chorismate-derived pathways. PMID- 19189965 TI - Inhibiting the calcineurin-NFAT (nuclear factor of activated T cells) signaling pathway with a regulator of calcineurin-derived peptide without affecting general calcineurin phosphatase activity. AB - Calcineurin phosphatase plays a crucial role in T cell activation. Dephosphorylation of the nuclear factors of activated T cells (NFATs) by calcineurin is essential for activating cytokine gene expression and, consequently, the immune response. Current immunosuppressive protocols are based mainly on calcineurin inhibitors, cyclosporine A and FK506. Unfortunately, these drugs are associated with severe side effects. Therefore, immunosuppressive agents with higher selectivity and lower toxicity must be identified. The immunosuppressive role of the family of proteins regulators of calcineurin (RCAN, formerly known as DSCR1) which regulate the calcineurin-NFAT signaling pathway, has been described recently. Here, we identify and characterize the minimal RCAN sequence responsible for the inhibition of calcineurin-NFAT signaling in vivo. The RCAN-derived peptide spanning this sequence binds to calcineurin with high affinity. This interaction is competed by a peptide spanning the NFAT PXIXIT sequence, which binds to calcineurin and facilitates NFAT dephosphorylation and activation. Interestingly, the RCAN-derived peptide does not inhibit general calcineurin phosphatase activity, which suggests that it may have a specific immunosuppressive effect on the calcineurin-NFAT signaling pathway. As such, the RCAN-derived peptide could either be considered a highly selective immunosuppressive compound by itself or be used as a new tool for identifying innovative immunosuppressive agents. We developed a low throughput assay, based on the RCAN1-calcineurin interaction, which identifies dipyridamole as an efficient in vivo inhibitor of the calcineurin-NFAT pathway that does not affect calcineurin phosphatase activity. PMID- 19189966 TI - A Cytosolic Homomerization and a Modulatory Domain within STIM1 C Terminus Determine Coupling to ORAI1 Channels. AB - In immune cells, generation of sustained Ca(2+) levels is mediated by the Ca(2+) release-activated Ca(2+) (CRAC) current. Molecular key players in this process comprise the stromal interaction molecule 1 (STIM1) that functions as a Ca(2+) sensor in the endoplasmic reticulum and ORAI1 located in the plasma membrane. Depletion of endoplasmic reticulum Ca(2+) stores leads to STIM1 multimerization into discrete puncta, which co-cluster with ORAI1 to couple to and activate ORAI1 channels. The cytosolic C terminus of STIM1 is sufficient to activate ORAI1 currents independent of store depletion. Here we identified an ORAI1-activating small fragment (OASF, amino acids 233-450/474) within STIM1 C terminus comprising the two coiled-coil domains and additional 50-74 amino acids that exhibited enhanced interaction with ORAI1, resulting in 3-fold increased Ca(2+) currents. This OASF, similar to the complete STIM1 C terminus, displayed the ability to homomerize by a novel assembly domain that occurred subsequent to the coiled-coil domains. A smaller fragment (amino acids 233-420) generated by a further deletion of 30 amino acids substantially reduced the ability to homomerize concomitant to a loss of coupling to as well as activation of ORAI1. Extending OASF by 35 amino acids (233-485) did not alter homomerization but substantially decreased efficiency in coupling to and activation of ORAI1. Expressing OASF in rat basophilic leukemia (RBL) mast cells demonstrated its enhanced plasma membrane targeting associated with 2.5-fold larger CRAC currents in comparison with the complete STIM1 C terminus. In aggregate, we have identified two cytosolic key regions within STIM1 C terminus that control ORAI1/CRAC activation: a homomerization domain indispensable for coupling to ORAI1 and a modulatory domain that controls the extent of coupling to ORAI1. PMID- 19189967 TI - Proteolytic release of the intramolecular chaperone domain confers processivity to endosialidase F. AB - Endosialidases (endoNs), as identified so far, are tailspike proteins of bacteriophages that specifically bind and degrade the alpha2,8-linked polysialic acid (polySia) capsules of their hosts. The crystal structure solved for the catalytic domain of endoN from coliphage K1F (endoNF) revealed a functional trimer. Folding of the catalytic trimer is mediated by an intramolecular C terminal chaperone domain. Release of the chaperone from the folded protein confers kinetic stability to endoNF. In mutant c(S), the replacement of serine 911 by alanine prevents proteolysis and generates an enzyme that varies in activity from wild type. Using soluble polySia as substrate a 3-times higher activity was detected while evaluation with immobilized polySia revealed a 190 fold reduced activity. Importantly, activity of c(S) did not differ from wild type with tetrameric sialic acid, the minimal endoNF substrate. Furthermore, we show that the presence of the chaperone domain in c(S) destabilizes binding to polySia in a similar way as did selective disruption of a polySia binding site in the stalk domain. The improved catalytic efficiency toward soluble polySia observed in these mutants can be explained by higher dissociation and association probabilities, whereas inversely, an impaired processivity was found. The fact that endoNF is a processive enzyme introduces a new molecular basis to explain capsule degradation by bacteriophages, which until now has been regarded as a result of cooperative interaction of tailspike proteins. Moreover, knowing that release of the chaperone domain confers kinetic stability and processivity, conservation of the proteolytic process can be explained by its importance in phage evolution. PMID- 19189968 TI - Estrogen receptor beta as a mitochondrial vulnerability factor. AB - We recently demonstrated mitochondrial localization of estrogen receptor beta (ERbeta). We herein confirm the mitochondrial localization of ERbeta by the loss of mitochondrial ERbeta immunoreactivity in ERbeta knockdown cells. A phenotype change characterized as an increase in resistance to oxidative stressors is associated with ERbeta knockdown. ERbeta knockdown results in a lower resting mitochondrial membrane potential (Deltapsim) and increase in resistance to hydrogen peroxide-induced Deltapsim depolarization in both immortal hippocampal cells and primary hippocampal neurons. ERbeta knockdown cells maintained ATP concentrations despite insults that compromise ATP production and produce less mitochondrial superoxide under oxidative stress. Furthermore, similar mitochondrial phenotype changes were identified in primary hippocampal neurons derived from ERbeta knock-out mice. These data demonstrate that ERbeta is expressed in mitochondria and function as a mitochondrial vulnerability factor involved in Deltapsim maintenance, potentially through a mitochondrial transcription dependent mechanism. PMID- 19189969 TI - Kruppel-like factor 5 shows proliferation-specific roles in vascular remodeling, direct stimulation of cell growth, and inhibition of apoptosis. AB - Kruppel-like factor 5 (KLF5), originally isolated as a regulator of phenotypic modulation of vascular smooth muscle cells, induces pathological cell growth and is expressed in the neointima. Although induction of KLF5 up-regulates growth factors like platelet-derived growth factor-A chain, how KLF5 actually contributes to vascular remodeling, notably its direct effects on cell proliferation, had been poorly clarified. To investigate the effects of KLF5 on neointimal formation, we at first performed adenoviral overexpression of KLF5 to rats subjected to carotid balloon injury. Neointimal formation and proliferating cell nuclear antigen-positive rate were significantly increased at 14 days after injury in the KLF5-treated animals. At the cellular level, overexpression of KLF5 also resulted in markedly increased cell proliferation and cell cycle progression. As a molecular mechanism, we showed that KLF5 directly bound to the promoter and up-regulated gene expression of cyclin D1, as well as showing specific transactivation of cyclins and cyclin-dependent kinase inhibitors in cardiovascular cells. Conversely, knockdown of KLF5 by RNA interference specifically down-regulated cyclin D1 and impaired vascular smooth muscle cell proliferation. Furthermore, KLF5 attenuated cleavage of caspase-3 under conditions of apoptotic stimulation. Moreover, KLF5-administered animals exhibited a significant decrease in terminal deoxynucleotidyltransferase-mediated dUTP nick end-labeling-positive cells in the medial layer, suggesting inhibition of apoptosis in the early phase after denudation. These findings collectively suggest that KLF5 plays a central role in cardiovascular pathologies through direct and specific stimulation of cell growth as well as inhibition of apoptosis. PMID- 19189970 TI - Intramembrane proteolysis by signal peptide peptidases: a comparative discussion of GXGD-type aspartyl proteases. AB - Intramembrane-cleaving proteases are required for reverse signaling and membrane protein degradation. A major class of these proteases is represented by the GXGD type aspartyl proteases. GXGD describes a novel signature sequence that distinguishes these proteases from conventional aspartyl proteases. Members of the family of the GXGD-type aspartyl proteases are the Alzheimer disease-related gamma-secretase, the signal peptide peptidases and their homologs, and the bacterial type IV prepilin peptidases. We will describe the major biochemical and functional properties of the signal peptide peptidases and their relatives. We then compare these properties with those of gamma-secretase and discuss common mechanisms but also point out a number of substantial differences. PMID- 19189971 TI - Intramembrane-cleaving proteases. PMID- 19189972 TI - Functional characterization of tlmK unveiling unstable carbinolamide intermediates in the tallysomycin biosynthetic pathway. AB - Tallysomycins (TLMs) belong to the bleomycin family of anticancer antibiotics. TLMs differ from bleomycins primarily by the presence of a 4-amino-4,6-dideoxy-l talose sugar attached to C-41 as part of a glycosylcarbinolamide. We previously proposed, on the basis of bioinformatics analysis of the tlm biosynthetic gene cluster from Streptoalloteichus hindustanus E465-94 ATCC 31158, that the tlmK gene is responsible for the attachment of this sugar moiety. We now report that inactivation of tlmK in S. hindustanus abolished TLM A and TLM B production, the resultant DeltatlmK mutant instead accumulated five new metabolites, and introduction of a functional copy of tlmK to the DeltatlmK mutant restored TLM A and TLM B production. Two major metabolites, TLM K-1 and TLM K-2, together with three minor metabolites, TLM K-3, TLM K-4, and TLM K-5, were isolated from the DeltatlmK mutant, and their structures were elucidated. These findings provide experimental evidence supporting the previous functional assignment of tlmK to encode a glycosyltransferase and unveil two carbinolamide pseudoaglycones as key intermediates in the TLM biosynthetic pathway. TlmK stabilizes the carbinolamide intermediates by glycosylating their hemiaminal hydroxyl groups, thereby protecting them from hydrolysis during TLM biosynthesis. In the absence of TlmK, the carbinolamide intermediates fragment to produce an amide TLM K-1 and aldehyde intermediates, which undergo further oxidative fragmentation to afford carboxylic acids TLM K-2, TLM K-3, TLM K-4, and TLM K-5. PMID- 19189973 TI - Critical roles of subunit NuoH (ND1) in the assembly of peripheral subunits with the membrane domain of Escherichia coli NDH-1. AB - The bacterial proton-translocating NADH:quinone oxidoreductase (NDH-1) consists of two domains, a peripheral arm and a membrane arm. NuoH is a counterpart of ND1, which is one of seven mitochondrially encoded hydrophobic subunits, and is considered to be involved in quinone/inhibitor binding. Sequence comparison in a wide range of species showed that NuoH is comprehensively conserved, particularly with charged residues in the cytoplasmic side loops. We have constructed 40 mutants of 27 conserved residues predicted to be in the cytoplasmic side loops of Escherichia coli NuoH by utilizing the chromosomal DNA manipulation technique and investigated roles of these residues. Mutants of Arg(37), Arg(46), Asp(63), Gly(134), Gly(145), Arg(148), Glu(220), and Glu(228) showed low deamino-NADH K(3)Fe(CN)(6) reductase activity, undetectable NDH-1 in Blue Native gels, low contents of peripheral subunits (especially NuoB and NuoCD) bound to the membranes, and a significant loss of the membrane potential and proton-pumping function coupled to deamino-NADH oxidation. The results indicated that these conserved residues located in the cytoplasmic side loops are essential for the assembly of the peripheral subunits with the membrane arm. Implications for the involvement of NuoH (ND1) in maintaining the structure and function of NDH-1 are discussed. PMID- 19189974 TI - GLI1 is a direct transcriptional target of EWS-FLI1 oncoprotein. AB - Ewing sarcoma family of tumors (ESFT) is an undifferentiated neoplasm of the bone and soft tissue. ESFT is characterized by a specific chromosomal translocation occurring between chromosome 22 and (in most cases) chromosome 11, which generates an aberrant transcription factor, EWS-FLI1. The function of EWS-FLI1 is essential for the maintenance of ESFT cell survival and tumorigenesis. The Hedgehog pathway is activated in several cancers. Oncogenic potential of the Hedgehog pathway is mediated by increasing the activity of the GLI family of transcription factors. Recent evidence suggests that EWS-FLI1 increases expression of GLI1 by an unknown mechanism. Our data from chromatin immunoprecipitation and promoter reporter studies indicated GLI1 as a direct transcriptional target of EWS-FLI1. Expression of EWS-FLI1 in non-ESFT cells increased GLI1 expression and GLI-dependent transcription. We also detected high levels of GLI1 protein in ESFT cell lines. Pharmacological inhibition of GLI1 protein function decreased proliferation and soft agar colony formation of ESFT cells. Our results establish GLI1 as a direct transcriptional target of EWS-FLI1 and suggest a potential role for GLI1 in ESFT tumorigenesis. PMID- 19189976 TI - Genetic network inference as a series of discrimination tasks. AB - MOTIVATION: Genetic network inference methods based on sets of differential equations generally require a great deal of time, as the equations must be solved many times. To reduce the computational cost, researchers have proposed other methods for inferring genetic networks by solving sets of differential equations only a few times, or even without solving them at all. When we try to obtain reasonable network models using these methods, however, we must estimate the time derivatives of the gene expression levels with great precision. In this study, we propose a new method to overcome the drawbacks of inference methods based on sets of differential equations. RESULTS: Our method infers genetic networks by obtaining classifiers capable of predicting the signs of the derivatives of the gene expression levels. For this purpose, we defined a genetic network inference problem as a series of discrimination tasks, then solved the defined series of discrimination tasks with a linear programming machine. Our experimental results demonstrated that the proposed method is capable of correctly inferring genetic networks, and doing so more than 500 times faster than the other inference methods based on sets of differential equations. Next, we applied our method to actual expression data of the bacterial SOS DNA repair system. And finally, we demonstrated that our approach relates to the inference method based on the S system model. Though our method provides no estimation of the kinetic parameters, it should be useful for researchers interested only in the network structure of a target system. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 19189977 TI - Systems biology metabolic modeling assistant: an ontology-based tool for the integration of metabolic data in kinetic modeling. AB - SUMMARY: We present Systems Biology Metabolic Modeling Assistant (SBMM Assistant), a tool built using an ontology-based mediator, and designed to facilitate metabolic modeling through the integration of data from repositories that contain valuable metabolic information. This software can be used for the visualization, design and management of metabolic networks; selection, integration and storage of metabolic information; and as an assistant for kinetic modeling. AVAILABILITY: SBMM Assistant for academic use is freely available at http://www.sbmm.uma.es. PMID- 19189975 TI - Meta-analysis of age-related gene expression profiles identifies common signatures of aging. AB - MOTIVATION: Numerous microarray studies of aging have been conducted, yet given the noisy nature of gene expression changes with age, elucidating the transcriptional features of aging and how these relate to physiological, biochemical and pathological changes remains a critical problem. RESULTS: We performed a meta-analysis of age-related gene expression profiles using 27 datasets from mice, rats and humans. Our results reveal several common signatures of aging, including 56 genes consistently overexpressed with age, the most significant of which was APOD, and 17 genes underexpressed with age. We characterized the biological processes associated with these signatures and found that age-related gene expression changes most notably involve an overexpression of inflammation and immune response genes and of genes associated with the lysosome. An underexpression of collagen genes and of genes associated with energy metabolism, particularly mitochondrial genes, as well as alterations in the expression of genes related to apoptosis, cell cycle and cellular senescence biomarkers, were also observed. By employing a new method that emphasizes sensitivity, our work further reveals previously unknown transcriptional changes with age in many genes, processes and functions. We suggest these molecular signatures reflect a combination of degenerative processes but also transcriptional responses to the process of aging. Overall, our results help to understand how transcriptional changes relate to the process of aging and could serve as targets for future studies. AVAILABILITY: http://genomics.senescence.info/uarrays/signatures.html. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 19189978 TI - VANO: a volume-object image annotation system. AB - Volume-object annotation system (VANO) is a cross-platform image annotation system that enables one to conveniently visualize and annotate 3D volume objects including nuclei and cells. An application of VANO typically starts with an initial collection of objects produced by a segmentation computation. The objects can then be labeled, categorized, deleted, added, split, merged and redefined. VANO has been used to build high-resolution digital atlases of the nuclei of Caenorhabditis elegans at the L1 stage and the nuclei of Drosophila melanogaster's ventral nerve cord at the late embryonic stage. AVAILABILITY: Platform independent executables of VANO, a sample dataset, and a detailed description of both its design and usage are available at research.janelia.org/peng/proj/vano. VANO is open-source for co-development. PMID- 19189979 TI - Lightweight comparison of RNAs based on exact sequence-structure matches. AB - MOTIVATION: Specific functions of ribonucleic acid (RNA) molecules are often associated with different motifs in the RNA structure. The key feature that forms such an RNA motif is the combination of sequence and structure properties. In this article, we introduce a new RNA sequence-structure comparison method which maintains exact matching substructures. Existing common substructures are treated as whole unit while variability is allowed between such structural motifs. Based on a fast detectable set of overlapping and crossing substructure matches for two nested RNA secondary structures, our method ExpaRNA (exact pattern of alignment of RNA) computes the longest collinear sequence of substructures common to two RNAs in O(H.nm) time and O(nm) space, where H << n.m for real RNA structures. Applied to different RNAs, our method correctly identifies sequence-structure similarities between two RNAs. RESULTS: We have compared ExpaRNA with two other alignment methods that work with given RNA structures, namely RNAforester and RNA_align. The results are in good agreement, but can be obtained in a fraction of running time, in particular for larger RNAs. We have also used ExpaRNA to speed up state-of-the-art Sankoff-style alignment tools like LocARNA, and observe a tradeoff between quality and speed. However, we get a speedup of 4.25 even in the highest quality setting, where the quality of the produced alignment is comparable to that of LocARNA alone. AVAILABILITY: The presented algorithm is implemented in the program ExpaRNA, which is available from our website (http://www.bioinf.uni-freiburg.de/Software). PMID- 19189980 TI - Association between the interleukin 23 receptor and ankylosing spondylitis is confirmed by a new UK case-control study and meta-analysis of published series. AB - OBJECTIVES: It has been shown previously that IL-23R variants are associated with AS. We conducted an extended analysis in the UK population and a meta-analysis with the previously published studies, in order to refine these IL-23R associations with AS. METHODS: The UK case-control study included 730 new cases and 1331 healthy controls. In the extended study, the 730 cases were combined with 1088 published cases. Allelic associations were analysed using contingency tables. In the meta-analysis, 3482 cases and 3150 controls from four different published studies and the new UK cases were combined. DerSimonian-Laird test was used to calculate random effects pooled odds ratios (ORs). RESULTS: In the UK case-control study with new cases, four of the eight SNPs showed significant associations, whereas in the extended UK study, seven of the eight IL-23R SNPs showed significant associations (P < 0.05) with AS, maximal with rs11209032 (P < 10(-5), OR 1.3), when cases with IBD and/or psoriasis were excluded. The meta analysis showed significant associations with all eight SNPs; the strongest associations were again seen not only with rs11209032 (P = 4.06 x 10(-9), OR approximately 1.2) but also with rs11209026 (P < 10(-10), OR approximately 0.6). CONCLUSIONS: IL-23R polymorphisms are clearly associated with AS, but the primary causal association(s) is(are) still not established. These polymorphisms could contribute either increased or decreased susceptibility to AS; functional studies will be required for their full evaluation. Additionally, observed stronger associations with SNPs rs11209026 and rs11465804 upon exclusion of IBD and/or psoriasis cases may represent an independent association with AS. PMID- 19189981 TI - Recruitment of cardiac parasympathetic activity: effects of clonidine on cardiac vagal motoneurones, pressure lability, and cardiac baroreflex slope in rats. AB - BACKGROUND: Association of low cardiac vagal activity and poor outcome is demonstrated in the cardiology setting. This has not been addressed in the postoperative setting. Cardiac vagal motoneurones (CVMs) in the brain stem generate sinus arrhythmia. They may reduce blood pressure (BP) variability ('pressure lability'). An alpha-2 agonist, clonidine, was administered to assess whether cardiac vagal activity could be recruited from a very low baseline activity, increase the sensitivity of the cardiac baroreflex and sinus arrhythmia, and reduce the pressure lability. METHODS: In ventilated anaesthetized rats, single-unit activity from antidromically identified CVMs was recorded. Given complex interactions within the cardiac ganglion, a peripherally acting beta-blocker, atenolol, was administered before clonidine. RESULTS: Atenolol 2 mg kg(-1) i.v. did not change systolic BP (SBP), CVM firing rate and slope of the cardiac baroreflex analysed at CVM (SBP-CVM unit activity relationship) level, or at the heart level (SBP-RR interval relationship) but evoked a significant bradycardia. In the presence of atenolol 2 mg kg(-1) h(-1), clonidine 10-100 microg kg(-1) i.v. evoked a significant reduction in SBP, a large increase of CVM firing rate from a very low base line [0.16 (sd 0.28) to 1.37 (1.21) spikes s(-1), n=7 cells], and increased the slope of the cardiac baroreflex analysed at the CVM level or at the heart level. sds of SBP were reduced, and that of RR interval was increased. CONCLUSIONS: Following peripheral beta-blockade, clonidine activated CVMs from a very low baseline, increased the slope of the cardiac baroreflex and sinus arrhythmia, and reduced pressure lability. PMID- 19189982 TI - Thoracic epidural analgesia in post-thoracotomy patients: comparison of three different concentrations of levobupivacaine and sufentanil. AB - BACKGROUND: Relative effects of dosage, volume and concentration of local anaesthetics used for postoperative thoracic epidural analgesia are still under debate. In this randomized, prospective, double-blinded study, we evaluated the incidence of side-effects such as changes in arterial pressure, postoperative nausea, vomiting, and pruritus in patients admitted for thoracic surgery during continuous thoracic epidural infusion using levobupivacaine and sufentanil mixture in three different volumes. METHODS: We studied 150 patients who underwent thoracotomy with a thoracic epidural catheter placed between T4 and T7. The patients were randomized into three groups which received 10 mg h(-1) of levobupivacaine at three different concentrations (0.5%, 0.25%, and 0.15%), in combination with sufentanil at 2.6 microg h(-1). Haemodynamic effects, pruritus, nausea, vomiting, sensory and motor block, pain score, additional analgesic requirement, sedation, and patient satisfaction were registered immediately after the surgical operation and on the first, second, and third postoperative days. RESULTS: We did not detect any differences in the incidence of side-effects such as changes in arterial pressure, and also postoperative nausea, vomiting, and pruritus. The three groups were also similar with regard to patient characteristics, sensory and motor block, pain score, analgesic rescue dose, sedation, and patient satisfaction. CONCLUSIONS: The same dose of a mixture of levobupivacaine and sufentanil administered in three different volumes and concentrations during continuous thoracic epidural infusion for thoracotomy provided an equal incidence of adverse haemodynamic effects, nausea, vomiting, or pruritus. PMID- 19189983 TI - Population pharmacokinetics of the two enantiomers of tramadol and O-demethyl tramadol after surgery in children. AB - BACKGROUND: Few data are available on the stereoselective pharmacokinetics of tramadol in children. The aim of this study was to develop a population pharmacokinetic model for the (+)- and (-)-enantiomers of tramadol and its O demethyl tramadol metabolite (M1) in children. METHODS: Twenty-five children (1-8 yr) were included in this study. Tramadol was administered after surgery by continuous infusion (loading dose, 2 mg kg(-1) i.v. over 10 min followed by continuous infusion of 8 mg kg(-1) over 24 h). If pain relief was inadequate, additional 1 mg kg(-1) i.v. bolus doses of tramadol were given over 10 min. A two compartment structural model was used with NONMEM. RESULTS: For both enantiomers of tramadol, weight was the only patient characteristic parameter showing significant covariate effects on clearance (CL). CL increased by 5.7-6.1 litre h( 1) between 8-12 and 13-16 kg, and by 2.4-3.3 litre h(-1) between 13-16 and 17-33 kg. The rate constants associated with the metabolite elimination [0.144 h(-1), (+)-M1 and 0.18 h(-1), (-)-M1] were smaller than the elimination rate constants of the parent drugs [0.243 h(-1), (+)-tramadol and 0.241 h(-1), (-)-tramadol], suggesting that the metabolite disposition was rate-limited by its elimination. The presence of two subpopulations of patients was suspected on the basis of the observed bimodal distributions of the AUC(M1)/AUC(tramadol) ratios. CONCLUSIONS: The results of this study combine relationships between tramadol CL and patient covariates that may be useful for dose adjustment. Polymorphism is likely to contribute to the interpatient variability observed in the AUC M1/AUC tramadol ratios. PMID- 19189984 TI - Does implicit memory during anaesthesia persist in children? AB - BACKGROUND: Recent studies suggest that implicit memory (especially perceptual implicit memory) persists during adequate general anaesthesia in adults. Studies in children, however, have failed to demonstrate implicit memory during general anaesthesia, possibly because of differences in methodological design. We therefore designed a prospective study with the aim of evaluating implicit memory in children undergoing general anaesthesia, using a perceptual memory test based on the mere exposure effect, previously tested in a control group. METHODS: Twelve infrequent neutral words were played 12 times in a random sequence via headphones to 36 children aged 8-12 yr during elective or emergency surgery. The children were not premedicated, and general anaesthesia was maintained with isoflurane. The word presentation started immediately after the surgical incision. Within 36 h after the stimulus presentation, the memory was assessed by using a forced-choice preference judgement task. Time constraint and word deterioration with a low-pass filter were used to prevent the subjects from utilizing intentional retrieval. The implicit memory score was obtained by calculating the proportion of target words preferred, which was compared with the chance level (0.5). RESULTS: The percentage of correct responses given by the children was comparable with the chance level. The memory score was mean (sd) 0.48 (0.16) (95% CI 0.43-0.53). CONCLUSIONS: The use of a perceptual implicit memory test based on the mere exposure procedure in children failed to reveal any evidence of implicit memory under general anaesthesia. PMID- 19189985 TI - Nicotinic receptor-evoked hippocampal norepinephrine release is highly sensitive to inhibition by isoflurane. AB - BACKGROUND: Inhaled anaesthetics (IAs) produce multiple dose-dependent behavioural effects including amnesia, hypnosis, and immobility in response to painful stimuli that are mediated by distinct anatomical, cellular, and molecular mechanisms. Amnesia is produced at lower anaesthetic concentrations compared with hypnosis or immobility. Nicotinic acetylcholine receptors (nAChRs) modulate hippocampal neural network correlates of memory and are highly sensitive to IAs. Activation of hippocampal nAChRs stimulates the release of norepinephrine (NE), a neurotransmitter implicated in modulating hippocampal synaptic plasticity. We tested the hypothesis that IAs disrupt hippocampal synaptic mechanisms critical to memory by determining the effects of isoflurane on NE release from hippocampal nerve terminals. METHODS: Isolated nerve terminals prepared from adult male Sprague-Dawley rat hippocampus were radiolabelled with [(3)H]NE and either [(14)C]GABA or [(14)C]glutamate and superfused at 37 degrees C. Release evoked by a 2 min pulse of 100 microM nicotine or 5 microM 4-aminopyridine was evaluated in the presence or absence of isoflurane and/or selective antagonists. RESULTS: Nicotine-evoked NE release from rat hippocampal nerve terminals was nAChR- and Ca(2+)-dependent, involved both alpha7 and non-alpha7 subunit-containing nAChRs, and was partially dependent on voltage-gated Na(+) channel activation based on sensitivities to various antagonists. Isoflurane inhibited nicotine-evoked NE release (IC(50)=0.18 mM) more potently than depolarization-evoked NE release (IC(50)=0.27 mM, P=0.014), consistent with distinct presynaptic mechanisms of IA action. CONCLUSIONS: Inhibition of hippocampal nAChR-dependent NE release by subanaesthetic concentrations of isoflurane supports a role in IA-induced amnesia. PMID- 19189986 TI - Ketamine has no effect on bispectral index during stable propofol-remifentanil anaesthesia. AB - BACKGROUND: Ketamine 0.15-1 mg kg(-1) decreases postoperative morphine consumption, but 0.5 mg kg(-1) is associated with an increase in the bispectral index (BIS) values that can lead to an overdose of hypnotic agents. The purpose of our investigation was to study the effect of ketamine 0.2 mg kg(-1) administered over a 5 min period on the BIS during stable target-controlled infusion (TCI) propofol-remifentanil general anaesthesia. METHODS: Thirty ASA I or II patients undergoing abdominal laparoscopic surgery were included in this double-blind, randomized study. Anaesthesia was induced and maintained with a TCI of propofol and remifentanil. After 5 min of steady-state anaesthesia (BIS at 40) without surgical stimulation, patients received either an infusion of ketamine 0.2 mg kg(-1) or normal saline. The test drug was infused over 5 min. Standard parameters and BIS values were recorded every minute until 15 min post-infusion. RESULTS: The baseline mean (sd) value for the BIS was 37 (6.5) for the ketamine group and 39 (8.2) for the placebo group. The highest mean BIS value during the recording period was 41.5 (8.7) for the ketamine group and 40.1 (8.9) for the placebo group. BIS values were not statistically different between the groups (P=0.62); there was no significant change over time (P=0.65) with no group-time interaction (P=0.55). CONCLUSIONS: Under stable propofol and remifentanil TCI anaesthesia, a slow bolus infusion of ketamine 0.2 mg kg(-1) administered over a 5 min period did not increase the BIS value over the next 15 min. PMID- 19189987 TI - Influence of an increased intracranial pressure on cerebral and systemic haemodynamics during endoscopic neurosurgery: an animal model. AB - BACKGROUND: During endoscopic neurosurgery, direct mechanical stimulation of the brain by the endoscope and increased intracranial pressure (ICP) caused by the continuous rinsing can induce potentially lethal haemodynamic reflexes, brain ischaemia, and excessive fluid resorption. METHODS: In a newly presented rat model of endoscopic neurosurgery, stereotactic access to the cerebrospinal fluid was secured and the ICP was increased by controlled infusion until complete suppression of the cerebral perfusion pressure (CPP). The haematocrit (Hct) level was determined before and after the procedure. During the whole procedure, invasive arterial pressure, ICP, and heart rate were continuously recorded and evaluated in a subsequent offline analysis. After the procedure, the animals were allowed to recover and 7 days later they were killed for histological examination. RESULTS: Suppression of the CPP resulted in a severe hypertension combined with tachycardia or mild bradycardia. The Hct decreased from 41 to 35 over the minutes of CPP suppression. After cessation of the infusion, the ICP decreased to 37% of the plateau pressure within 2.5 s. In the first few minutes after restoration of normal ICP, five animals died because of pulmonary oedema. CONCLUSIONS: Upon complete suppression of the CPP, an obvious hypertension developed, often together with tachycardia, but no severe bradycardia. At high ICP levels, we observed an important translocation of irrigation fluid to the vascular space. Fatality was not caused by ischaemia or arrhythmia but due to pulmonary oedema. PMID- 19189988 TI - Alstonine as an antipsychotic: effects on brain amines and metabolic changes. AB - Managing schizophrenia has never been a trivial matter. Furthermore, while classical antipsychotics induce extrapyramidal side effects and hyperprolactinaemia, atypical antipsychotics lead to diabetes, hyperlipidaemia, and weight gain. Moreover, even with newer drugs, a sizable proportion of patients do not show significant improvement. Alstonine is an indole alkaloid identified as the major component of a plant-based remedy used in Nigeria to treat the mentally ill. Alstonine presents a clear antipsychotic profile in rodents, apparently with differential effects in distinct dopaminergic pathways. The aim of this study was to complement the antipsychotic profile of alstonine, verifying its effects on brain amines in mouse frontal cortex and striatum. Additionally, we examined if alstonine induces some hormonal and metabolic changes common to antipsychotics. HPLC data reveal that alstonine increases serotonergic transmission and increases intraneuronal dopamine catabolism. In relation to possible side effects, preliminary data suggest that alstonine does not affect prolactin levels, does not induce gains in body weight, but prevents the expected fasting-induced decrease in glucose levels. Overall, this study reinforces the proposal that alstonine is a potential innovative antipsychotic, and that a comprehensive understanding of its neurochemical basis may open new avenues to developing newer antipsychotic medications. PMID- 19189989 TI - Acupuncture and auricular acupressure in relieving menopausal hot flashes of bilaterally ovariectomized chinese women: a randomized controlled trial. AB - The objective of this study is to explore the effects of acupuncture and auricular acupressure in relieving menopausal hot flashes of bilaterally ovariectomized Chinese women. Between May 2006 and March 2008, 46 bilaterally ovariectomized Chinese women were randomized into an acupuncture and auricular acupressure group (n = 21) and a hormone replacement therapy (HRT) group (Tibolone, n = 25). Each patient was given a standard daily log and was required to record the frequency and severity of hot flashes and side effects of the treatment felt daily, from 1 week before the treatment started to the fourth week after the treatment ended. The serum levels of follicle stimulating hormone (FSH), LH and E(2) were detected before and after the treatment. After the treatment and the follow-up, both the severity and frequency of hot flashes in the two groups were relieved significantly when compared with pre-treatment (P < .05). There was no significant difference in the severity of hot flashes between them after treatment (P > .05), while after the follow-up, the severity of hot flashes in the HRT group was alleviated more. After the treatment and the follow up, the frequency of menopausal hot flashes in the HRT group was reduced more (P < .05). After treatment, the levels of FSH decreased significantly and the levels of E(2) increased significantly in both groups (P < .05), and they changed more in the HRT group (P < .05). Acupuncture and auricular acupressure can be used as alternative treatments to relieve menopausal hot flashes for those bilaterally ovariectomized women who are unable or unwilling to receive HRT. PMID- 19189990 TI - The effect of Medicare Part D coverage on drug use and cost sharing among seniors without prior drug benefits. AB - This study evaluates the effect of Medicare Part D among seniors who previously lacked drug coverage, using time-trend analyses of patient-level dispensing data from three pharmacy chains. Of 114,766 seniors without drug benefits, 55 percent initiated drug insurance under Part D. After the penalty-free Part D enrollment period, use of statins, clopidogrel, and proton pump inhibitors stabilized at levels ranging from 11 percent to 37 percent above the trend that would have been expected if Part D had not been implemented. Patients reaching the Part D coverage gap (12 percent) experienced a decrease in essential medication use ranging from 5.7 percentage points per month for warfarin to 6.3 percentage points for statins. PMID- 19189991 TI - The effects of the coverage gap on drug spending: a closer look at Medicare Part D. AB - We calculated prescription drug usage in two groups of Medicare beneficiaries: employer group with no coverage gap, and individual Part D group with no coverage or some generic drug coverage in the coverage gap. Among those with employer coverage, 40 percent reached the doughnut hole, compared with 25 percent of those without such coverage. Overall, 5 percent went through the doughnut hole to reach the catastrophic coverage level. Those lacking coverage in the doughnut hole reduced their drug use by 14 percent; those with generic coverage reduced their use by 3 percent. Coverage of generic drugs with a $0-$10 copayment in the doughnut hole could be financed by, at most, a six-to-nine-percentage-point increase in initial coinsurance. PMID- 19189992 TI - Superficial ovarian cortex vascularization is inversely related to the follicle reserve in normal cycling ovaries and is increased in polycystic ovary syndrome. AB - BACKGROUND: The superficial ovarian cortex constitutes the micro-environment where resting and early growing follicles reside. As small follicles do not possess an independent capillary network, both their survival and early growth depend on their proximity to the cortical vessels. Little is known about the possible changes in superficial ovarian cortex vascularization in normal women throughout reproductive life or in pathological conditions such as polycystic ovary syndrome (PCOS) involving abnormal early follicle growth. We studied the vascularization of the superficial and deep cortical stroma (DCS) in normal cycling ovaries from 21 to 50 years of age and in infertile women with PCOS. METHODS: We used archival ovarian samples and specific CD34 immunostaining to determine blood vessel density and to analyse correlation with age and with the ovarian follicle reserve. RESULTS: Normal cycling ovaries showed an age-related increase in the superficial cortical stroma vascularization that was inversely correlated with the density of small (primordial and primary) follicles. In contrast, blood vessel density in the DCS significantly decreased in women aged >or=40 years. Ovaries from PCOS showed a 2-fold increase in blood vessel density in both superficial cortical stroma and DCS with respect to age-matched controls. CONCLUSIONS: The increased vascularization of the superficial cortical stroma in normal ovaries in relation to age and in ovaries from PCOS could have profound effects on cortical metabolic rate, primordial follicle survival/activation and early follicle growth, and may underline changes in follicle dynamics in mid-aged women and in PCOS. PMID- 19189993 TI - Providing infertility treatment in resource-poor countries. AB - Recently, several initiatives were started to introduce medically assisted reproduction in developing countries. Infertility is a major problem in these countries and causes extensive social and psychological suffering. This article analyses the main ethical arguments pro and contra the provision of infertility treatment in resource-poor countries. It is concluded that infertility treatment should be part of an integrated reproductive care programme including family planning and motherhood care. Education, empowerment of women and economic prosperity are the most effective solutions to most problems related to both population growth and infertility. Simultaneously, investments in low-cost interventions are justified. PMID- 19189994 TI - Frequency and characteristics of endometrial carcinoma and atypical hyperplasia detected on routine infertility investigations in young women: a report of six cases. AB - Infertility patients are known to be at increased risk of endometrial carcinoma (EC) and atypical hyperplasia (AH). However, the incidence and clinical features of EC and AH in these patients remain to be clarified. In this study, we examined the rate at which a routine infertility workup revealed EC/AH and investigated the clinicopathological features of EC/AH detected in this way. Among patients diagnosed with EC or AH at the Jichi Medical University Hospital between the 10 year period from 1997 to 2006, six patients were referred from Tochigi Central Clinic, a specialized infertility facility. We report the clinicopathological features of these patients and calculate the incidence of EC/AH in patients who underwent infertility investigations at Tochigi Central Clinic. All six patients were younger than 40 and had early stage disease (final diagnosis: EC stage IA: 3, EC stage IB: 1, AH: 2). A total of 19 826 patients underwent routine infertility investigations at Tochigi Central Clinic during the same period. The incidence of EC/AH detected from these investigations was 0.03% (6/19 826) and that of EC was 0.02% (4/19 826): 5-10 times higher than the overall incidence in Japanese women of the same age. Routine infertility investigations may provide an opportunity to examine the corpus uteri of young women in whom examination is otherwise limited, contributing to the early detection of EC/AH. PMID- 19189995 TI - Dopamine agonist administration causes a reduction in endometrial implants through modulation of angiogenesis in experimentally induced endometriosis. AB - BACKGROUND: Implantation of a retrogradely-shed endometrium during menstruation requires an adequate blood supply. The endometrium has angiogenic potential, and endometriotic lesions grow in areas with a rich vascularization, suggesting that angiogenesis is a prerequisite for endometriosis development. Targeting vascular endothelial growth factor (VEGF) leads to an inhibition of endometriosis. Dopamine and its agonists, such as cabergoline (Cb2), promote VEGF receptor-2 (VEGFR-2) endocytosis in endothelial cells, preventing VEGF-VEGFR-2 binding and reducing neoangiogenesis. The aim of this study was to evaluate the anti angiogenic properties of Cb2 on growth of established endometriosis lesions and investigate the molecular mechanisms by which Cb2 exerts the anti-angiogenic effect. METHODS: Human endometrium fragments were implanted in female nude mice peritoneum, and mice were treated with vehicle, 0.05 or 0.1 mg/kg/day oral Cb2 for 14 days. After treatment, the implants were processed to assess proliferative activity, neoangiogenesis, VEGFR-2 phosphorylation and angiogenic gene expression. RESULTS: A significant decrease in the percentage of active endometriotic lesions (P < 0.05) and cellular proliferation index (P < 0.001) was found with Cb2 treatment. Neoangiogenesis was reduced by Cb2 treatment, as observed at gross morphological level and by significant changes in gene expression. The degree of VEGFR-2 phosphorylation was significantly lower in Cb2 treated animals than controls. CONCLUSIONS: Cb2 treatment in experimental endometriosis has an anti-angiogenic effect acting through VEGFR-2 activation. These findings support the testing of dopamine agonists as a novel therapeutic approach to peritoneal endometriosis in humans. PMID- 19189997 TI - Apligraf in the treatment of neuropathic diabetic foot ulcers. AB - This study compared the efficacy and safety of Apligraf (Organogenesis, Inc., Canton, MA) in combination with standard therapy versus standard therapy alone in the treatment of neuropathic diabetic foot ulcers. Efficacy was assessed by time to complete wound healing (by 12 weeks) and incidence of complete wound closure (at 12 weeks). This was an international multi-center, randomized, controlled study. Patients were eligible for entry into the study if the following criteria were met: type 1 or type 2 diabetes mellitus, age 18 to 80 years, adequate glycemic control, and the presence of a full-thickness neuropathic ulcer for at least 2 weeks prior to the initial screening visit. Following the 2-week screening period, the 2 treatment groups received standard ulcer care consistent with international treatment guidelines that comprised sharp debridement, saline moistened dressings, and a non-weight bearing regimen. There were 106 subjects screened for enrollment, 82 randomized to the treatment groups, and 72 treated (33 Apligraf subjects and 39 standard therapy subjects) before the study was terminated. Kaplan-Meier curves indicated a trend for shorter time to complete wound healing in the Apligraf group compared with the standard therapy group (p = .059; log-rank test). The median time to healing was 84 days in the Apligraf group, whereas no median time to healing could be determined for the standard therapy group because <50% of the standard therapy subjects healed. By 12 weeks, 51.5% (17/33) Apligraf subjects had achieved complete wound closure compared with 26.3% (10/38) of standard therapy subjects (p = .049; Fisher's exact test). Even though the study was halted prematurely, this study suggested that the use of Apligraf resulted in a higher incidence of wound closure by 12 weeks. PMID- 19189996 TI - Sweet liking phenotype, alcohol craving and response to naltrexone treatment in alcohol dependence. AB - AIMS: To investigate the relationship between the sweet liking/sweet disliking phenotype (a putative probe of brain opioid function), craving for alcohol and response to treatment with naltrexone in individuals with alcohol dependence. METHODS: Forty individuals with alcohol dependence were enrolled in a 12-week open-label study of 50 mg of naltrexone with four sessions of motivational enhancement therapy. Prior to treatment, individuals completed a sweet preference test and the Penn Alcohol Craving Scale. Subjects were categorized as sweet liking (SL), n = 15, or sweet disliking (SDL), n = 25, via a standard sweet tasting paradigm. The sweet tasting results were blinded to the subjects and to treatment staff. SL status, pretreatment craving and their interaction were examined as predictors of frequency of abstinent days and heavy drinking days during treatment with naltrexone. RESULTS: SL and SDL subjects achieved similar reductions in percent heavy drinking days with treatment. During treatment, SDL subjects had 48% abstinent days compared to 30% for SL subjects (P = 0.034). Pretreatment craving did not predict % heavy drinking days or % abstinent days. An interaction effect was found between the SL/SDL phenotype and pretreatment craving such that SL subjects with high craving demonstrated higher rates of percent abstinent days whereas SDL subjects with high craving demonstrated lower rates of percent abstinent days, P < 0.001. CONCLUSIONS: These findings indicate that the SL/SDL phenotype may predict variation in response to naltrexone and/or counseling treatment. Furthermore, the SL/SDL phenotype may interact with craving to provide a more robust prediction of outcome with naltrexone or counseling. PMID- 19189998 TI - Warthin tumor of the upper lip: an unusual location of a benign salivary gland tumor. AB - Warthin tumor (papillary cystadenoma lymphomatosum) is a benign salivary gland tumor involving almost exclusively the parotid gland. The lip is a very unusual location for this type of tumor, which develops only rarely in minor salivary glands. The case of 42-year-old woman with Warthin tumor arising in minor salivary glands of the upper lip is reported. PMID- 19189999 TI - Reliability (internal consistency) of the job content questionnaire on job stress among office workers of a multinational company in Kuala Lumpur. AB - This small, cross-sectional study assessed the reliability of 3 scales from the Job Content Questionnaire (JCQ)-decision latitude, psychological job demand, and social support-in a group of office workers in a multinational company in Kuala Lumpur. A universal sample of 30 white-collar workers from a department of the company self-administered the English version of the JCQ comprising 21 core items selected from the full recommended version of 49 items on-site. Reliability (internal consistency) was evaluated using Cronbach's alpha coefficients for each scale. Corrected item-total correlation was presented for each and every item. Cronbach's alpha coefficients were acceptable for decision latitude (.76) and social support (.79) but slightly lower for psychological job demand (.64). Values for all item-total correlations for all 3 scales were greater than .3. In conclusion, this study suggests that the JCQ is a reliable scale for assessing job stress in this group of workers. PMID- 19190000 TI - Women's autonomy in decision making for health care in South Asia. AB - This article aims to discuss women's autonomy in decision making on health care, and its determinants in 3 South Asian countries, using nationally representative surveys. Women's participation either alone or jointly in household decisions on their own health care was considered as an indicator of women's autonomy in decision making. The results revealed that decisions of women's health care were made without their participation in the majority of Nepal (72.7%) and approximately half of Bangladesh (54.3%) and Indian (48.5%) households. In Sri Lanka, decision making for contraceptive use was a collective responsibility in the majority (79.7%). Women's participation in decision making significantly increased with age, education, and number of children. Women who were employed and earned cash had a stronger say in household decision making than women who did not work or worked not for cash. Rural and poor women were less likely to be involved in decision making than urban or rich women. PMID- 19190001 TI - Alcohol consumption by older adults in central and southern Japan. AB - This study ascertained the level of alcohol intake and alcoholic beverages consumed by Japanese older adults. Persons aged 55 to 75 years residing in central and southern Japan were recruited and interviewed face-to-face on their habitual alcohol consumption. Among the 577 (359 men and 218 women) participants from 10 districts/prefectures, 60.5% (75.5% for men and 35.7% for women) regularly drank alcoholic beverages on at least a monthly basis. Beer was the most preferred beverage (45.2%), followed by shochu (19.8%) and sake (16.1%). The mean alcohol consumption was 22.2 g/day (95% confidence interval 19.3-25.2) overall, but drinkers had a much higher mean intake of 36.6 g/day (95% CI 32.4 40.8). Moreover, it is alarming that 25.5% of male drinkers were heavy drinkers consuming more than 60 g of alcohol on average per day. Alcohol control measures should be developed to curtail the excessive drinking by older adults. PMID- 19190002 TI - Costing dental restorations in public sector dental clinics. AB - The objective of this study is to share cost analysis methodology and to obtain cost estimates for posterior restorations in public sector dental clinics. Two urban and 2 rural dental clinics in Selangor state were selected. Only cases of 1 posterior restoration per visit by dental officers were included over 6 months. One capsulated amalgam type, 1 capsulated tooth-colored, and 1 non-capsulated tooth-colored material were selected. A clinical pathway form was formulated to collect data per patient. Annual capital and recurrent expenditures were collected per clinic. The mean cost of an amalgam restoration was RM 30.96 (sdRM 7.86); and tooth-colored restorations ranged from RM 33.00 (sdRM 8.43) to RM 41.10 (sdRM 10.61). Wherein 1 USD = RM 2.8. Restoration costs were 35% to 55% higher in clinics in rural areas than in urban areas. The findings demonstrate economy of scale for clinic operation and restoration costs with higher patient load. Costs per restoration were higher in rural than in urban dental clinics. More studies are recommended to address the dearth of dental costs data in Malaysia. PMID- 19190003 TI - Prevalence of obesity among adolescents (10 to 14 years) in Kuwait. AB - The purpose of this cross-sectional study was to find out the prevalence of obesity and overweight among intermediate school adolescents aged 10 to 14 years. The study comprised a multistage stratified random sample that included 5402 children (2657 males and 2745 females). They represent 12.7% of the total number of children between 10 and 14 years during the educational year 2005-2006. The weights and heights of adolescents were measured, from which the body mass index (BMI) was calculated, which is the weight in kilograms divided by the height in meters squared (kg/m(2)). BMI values higher than 95 percentile were accepted as being obese and those in between 85 and 94 percentile were accepted as overweight. Dietary intake was assessed by the investigators using food exchange lists designed by American Diabetic Association and physical fitness was measured by modified Harvard step test. Data regarding monthly income of the chosen sample were collected from parents of those children. The overall prevalence of overweight and obesity in adolescent Kuwaiti children aged 10 to 14 years was 30.7% and 14.6%, respectively. The overall prevalence of overweight and obesity among males was 29.3% and 14.9%, respectively (P < .001) and the prevalence of overweight and obesity among females was 32.1% and 14.2%, respectively (P < .001). High daily caloric intake by the obese and overweight children and physical inactivity was reported among the majority of them. Health education programs should be conducted to control this syndrome in order to prevent future risk of obesity-related disease, and physical activity programs should be incorporated in the schools. Any management plan for overweight and obese children should include 3 major components: diets, exercise, and family-based behavior and they should not be placed on restrictive diets because adequate calories are needed for proper growth. PMID- 19190004 TI - A way to restore British paediatricians' engagement with child protection. PMID- 19190005 TI - Insulin injections in schools. PMID- 19190006 TI - BCG protects toddlers during a tuberculosis outbreak. AB - In 2007, an outbreak of tuberculosis occurred in a toddler population attending two child care centres in Cork, Ireland. Of 268 children exposed, 18 were eventually diagnosed with active tuberculosis. We present the initial clinical and radiographic characteristics of the active disease group. Mantoux testing was positive in only 66% of cases. All cases were either pulmonary or involved hilar adenopathy on chest radiograph; there were no cases of disseminated disease or meningitis. 24% of the exposed children had been previously vaccinated with BCG, and no case of active disease was found in this group (p = 0.016), suggesting a profound protective effect of BCG in this population. Our experience provides evidence supporting a protective effect of BCG against pulmonary disease in young children. PMID- 19190007 TI - Neonatal percutaneous central venous lines: fit to burst. AB - OBJECTIVE: To examine pressure changes in neonatal percutaneous central venous catheters under varying laboratory conditions and to quantify the risks of rupture in clinical practice. DESIGN: We tested 27-gauge polyurethane Premicath and 24-gauge silicone ECC (both Vygon, Norristown, PA) catheters. Burst pressures were determined by applying a slowly ramped pressure to catheters that were occluded at the tip. Flow-pressure relationships were defined by increasing flow rates through patent catheters from 5 to 499 ml/h. Pressure changes during the manual flushing of catheters were determined for patent and occluded catheters and with different syringe sizes. RESULTS: The mean burst pressure for polyurethane catheters (1730.8 kPa, 95% CI 1634.7 to 1826.8) was higher than for silicone catheters (275.6 kPa, 95% CI 240.4 to 310.8). Polyurethane catheters demonstrated an approximately fivefold greater margin of safety above manufacturer recommended operating pressures before burst compared to silicone catheters. Pressures remained at safe levels in both catheters over the range of flows generally used in neonatal practice. Hand-flushing of obstructed silicone catheters caused rupture in 5/6 silicone catheters tested, in comparison to 0/16 polyurethane catheters. CONCLUSIONS: Polyurethane central venous catheters have a greater pressure tolerance than silicone catheters and are less likely to rupture under experimental conditions. Obstructed silicone catheters rupture easily when flushed. Catheters were not tested in human infants. PMID- 19190008 TI - Don't throw your blood clots away: use of blood clot may improve sensitivity of PCR diagnosis in invasive aspergillosis. AB - BACKGROUND: The diagnosis of invasive aspergillosis (IA) remains challenging and frequently is not made until after death. Histopathological examination remains central to confirmation of diagnosis but often requires invasive procedures to obtain tissue for the examination. Detection of aspergillus DNA by quantitative PCR (qPCR) offers the potential for earlier diagnosis due to higher sensitivity, but PCR in clinical use is poorly reproducible, with different centres reporting variable results and often using different extraction and analytical methods. AIMS: To optimise the performance of aspergillus PCR as a diagnostic modality. METHODS: A rat inhalation model of invasive aspergillosis was used to optimise the methodology of diagnostic aspergillus PCR. Infected animals were terminally bled at 4 days post-infection; samples of EDTA blood, serum and the residual clot were pooled for subsequent analysis. DNA was extracted from each fraction using a variety of methods and an optimised qPCR reaction using an Aspergillus fumigatus primer set performed. RESULTS: Significantly more aspergillus DNA was detected from the clot than EDTA and serum samples. Enzymatic and mechanical pretreatment reduced the yield of fungal DNA. There was some evidence that the average Ct values were greater for the EZ1 BioRobot than the MagNA Pure automated extractor, but this did not reach statistical significance at the 5% level (p = 0.078). CONCLUSIONS: Automated extraction from the clot present in a blood sample will increase DNA yield and improve the diagnostic sensitivity of the test. PMID- 19190009 TI - Sentinel lymph node biopsy in patients with a needle core biopsy diagnosis of ductal carcinoma in situ: is it justified? AB - BACKGROUND: The incidence of ductal carcinoma in situ (DCIS) has increased markedly with the introduction of population-based mammographic screening. DCIS is usually diagnosed non-operatively. Although sentinel lymph node biopsy (SNB) has become the standard of care for patients with invasive breast carcinoma, its use in patients with DCIS is controversial. AIM: To examine the justification for offering SNB at the time of primary surgery to patients with a needle core biopsy (NCB) diagnosis of DCIS. METHODS: A retrospective analysis was performed of 145 patients with an NCB diagnosis of DCIS who had SNB performed at the time of primary surgery. The study focused on rates of SNB positivity and underestimation of invasive carcinoma by NCB, and sought to identify factors that might predict the presence of invasive carcinoma in the excision specimen. RESULTS: 7/145 patients (4.8%) had a positive sentinel lymph node, four macrometastases and three micrometastases. 6/7 patients had invasive carcinoma in the final excision specimen. 55/145 patients (37.9%) with an NCB diagnosis of DCIS had invasive carcinoma in the excision specimen. The median invasive tumour size was 6 mm. A radiological mass and areas of invasion <1 mm, amounting to "at least microinvasion" on NCB were predictive of invasive carcinoma in the excision specimen. CONCLUSIONS: SNB positivity in pure DCIS is rare. In view of the high rate of underestimation of invasive carcinoma in patients with an NCB diagnosis of DCIS in this study, SNB appears justified in this group of patients. PMID- 19190010 TI - Induced abortions and teenage births among asylum seekers in The Netherlands: analysis of national surveillance data. AB - BACKGROUND: Asylum seekers are assumed to be a vulnerable group with respect to sexual and reproductive health. The objective of this study was to quantify induced abortion and teenage birth indicators for this group. METHODS: The population comprised all female asylum seekers aged 15-49 in The Netherlands between September 2004 and August 2005. Information was collected about induced abortions from notification forms and electronic patient files. The central agency for the reception of asylum seekers provided population and birth data. RESULTS: Among asylum seekers the abortion rate (14.4/1000 women) and teenage birth rate (49.1/1000) were higher than average in The Netherlands (8.6/1000 and 5.8/1000). Great differences were found between subgroups. High abortion rates were seen among women who were pregnant on arrival or got pregnant in the first months after arrival at the reception facilities. Abortion and teenage birth rates were particularly high among asylum seekers aged 15-19 from specific parts of Africa and Asia. Abortion ratios were high among asylum seekers aged 30-49 from parts of Europe and Asia. Decreases in the abortion rate and teenage birth rate were observed as the length of stay increased. CONCLUSION: Abortions and teenage births were more common among asylum seekers than among the overall population of The Netherlands. Increased rates were a consequence of subgroups being at high risk. Abortion and teenage birth rates were very high among women who were pregnant on arrival or got pregnant in the first few months after arrival, but decreased as the length of stay increased. PMID- 19190011 TI - Starch storage in the stems of wheat plants: localization and temporal changes. AB - BACKGROUND AND AIMS: Carbohydrate temporarily accumulates in wheat stems during the early reproductive growth phase, predominantly as water soluble carbohydrate (WSC), and is subsequently remobilized during grain filling. Starch has also been reported as a minor storage carbohydrate component in wheat stems, but the details are lacking. METHODS: The accumulation and localization of starch in wheat stem and leaf sheath tissue over a developmental period from 6 d before anthesis to 35 d after anthesis was investigated. KEY RESULTS: The region of the peduncle enclosed by the flag-leaf sheath, and the penultimate internode were the main tissues identified as containing starch, in which the starch grains localized to the storage parenchyma cells. In contrast, the exposed peduncle lacked starch grains. Starch grains were also found in the flag-leaf and second leaf sheath. Plants grown in low-nitrogen conditions exhibited increased storage of both starch and WSC compared with plants grown in high-nitrogen supply. CONCLUSIONS: The major accumulation and decrease of starch occurred temporally independently to that for WSC, suggesting a different functional role for starch in wheat stems. Starch reutilization concomitant with peduncle growth, and the early development of the reproductive structures, suggested a role in provision of energy and/or carbon scaffolds for these growth processes. PMID- 19190012 TI - Colchicine for pericarditis: hype or hope? AB - Colchicine has been effectively used in the treatment of several inflammatory conditions, such as gouty attacks, serositis related to familial Mediterranean fever, Behcet syndrome, and more recently also in acute and recurrent pericarditis. Growing evidence has shown that the drug may be useful to treat an acute attack and may be a way to cope with the prevention of pericarditis in acute and recurrent cases and after cardiac surgery. Nevertheless, clinicians are often sceptical about the efficacy of the drug, and concerns have risen on possible side effects and tolerability. In this review, we analyse current evidence to support the use of the drug, as well as possible harms and risks related to drug interactions, reaching the conclusion that colchicine is safe and useful in recurrent pericarditis, if specific precautions are followed, although less evidence supports its use for the treatment of acute pericarditis, where colchicine remains optional and there is a need for further multicentre confirmatory studies. This paper also reviews specific dosing and precautions for the clinical use. PMID- 19190013 TI - Aldosterone blockade in patients with heart failure and a reduced left ventricular ejection fraction. PMID- 19190014 TI - Practice Pointer. "I need a note, doctor": dealing with requests for medical reports about patients. PMID- 19190016 TI - Westminster primary care trust introduces programme to prevent heart disease. PMID- 19190015 TI - Vulnerability and access to care for South Asian Sikh and Muslim patients with life limiting illness in Scotland: prospective longitudinal qualitative study. AB - OBJECTIVES: To examine the care experiences of South Asian Sikh and Muslim patients in Scotland with life limiting illness and their families and to understand the reasons for any difficulties with access to services and how these might be overcome. DESIGN: Prospective, longitudinal, qualitative design using in depth interviews. SETTING: Central Scotland. PARTICIPANTS: 25 purposively selected South Asian Sikh and Muslim patients, 18 family carers, and 20 key health professionals. RESULTS: 92 interviews took place. Most services struggled to deliver responsive, culturally appropriate care. Barriers to accessing effective end of life care included resource constrained services; institutional and, occasionally, personal racial and religious discrimination; limited awareness and understanding among South Asian people of the role of hospices; and difficulty discussing death. The most vulnerable patients, including recent migrants and those with poor English language skills, with no family advocate, and dying of non-malignant diseases were at particularly high risk of inadequate care. CONCLUSIONS: Despite a robust Scottish diversity policy, services for South Asian Sikh and Muslim patients with life limiting illness were wanting in many key areas. Active case management of the most vulnerable patients and carers, and "real time" support, from where professionals can obtain advice specific to an individual patient and family, are the approaches most likely to instigate noticeable improvements in access to high quality end of life care. Improving access to palliative care for all, particularly those with non-malignant illnesses, as well as focusing on the specific needs of ethnic minority groups, is required. PMID- 19190017 TI - Guideline seeks to clarify difference between palliative sedation and euthanasia. PMID- 19190018 TI - More medical students should be educated in the community, US report says. PMID- 19190020 TI - Gates foundation pledges new funds to fight neglected diseases. PMID- 19190022 TI - Pathophysiological and clinical relevance of simplified monitoring of nocturnal breathing disorders in heart failure patients. AB - AIMS: Nocturnal breathing disorders in the form of periodic breathing (PB) are very common in heart failure (HF) patients. There is an increasing interest in simple and affordable tools to screen patients and monitor these disorders at home on a long-term basis. We aimed to assess the pathophysiological and clinical relevance of a simplified method for monitoring of PB suitable to be self-managed by the patient at home. METHODS AND RESULTS: A night-time respiratory recording was performed in 397 optimally treated HF patients (age 60 +/- 11 years, NYHA class 2.4 +/- 0.6, left ventricular ejection fraction 29 +/- 7%) and the duration of PB (PB(Dur)) automatically computed. Patients were followed-up for 1 year and the prognostic value of PB(Dur) evaluated. In 45 patients, we assessed the association between PB(Dur) and severity of oxygen desaturations (number of desaturations >3%). Twenty six of the 397 patients died of cardiac causes. A PB(Dur) > or =2 h was significantly associated with an increased risk of cardiac death after adjustment for major clinical predictors [hazard ratio (95% CI): 3.5 (1.6-7.9), P = 0.002]. The correlation between PB(Dur) and severity of desaturations was 0.94 (P < 0.0001). CONCLUSION: Relevant pathophysiological and clinical information can be obtained from simplified monitoring of breathing disorders managed by the patient. These results provide new perspectives in the investigation of the clinical impact of abnormal breathing in HF patients. PMID- 19190023 TI - Prognostic value of plasma midregional pro-adrenomedullin and C-terminal-pro endothelin-1 in chronic heart failure outpatients. AB - AIMS: The identification of chronic heart failure (CHF) patients at high risk of adverse outcome remains a challenge. New peptides are emerging that may give additional information. In CHF patients, endothelin (ET) levels predict mortality risk. Adrenomedullin has been shown to predict mortality in ischaemic heart failure, but not in unselected or non-ischaemic CHF patients. Moreover, ADM and ET have never been assessed in one model. The aim of the present study was to assess the prognostic value of midregional-pro-adrenomedullin (MR-proADM) and C terminal-pro-endothelin-1 (CT-proET-1) in outpatients with CHF. METHODS AND RESULTS: We measured plasma MR-proADM and CT-proET-1 levels in 786 consecutive CHF outpatients and compared them with B-type natriuretic peptide (BNP) levels. At 24-month follow-up, 233 patients had died. A stepwise forward Cox regression model with age, sex, estimated glomerular filtration rate, NYHA > II, left ventricular ejection fraction (LVEF), MR-proADM, CT-proET-1, and BNP as possible predictors revealed that MR-proADM levels [hazard ratio (HR) = 1.77, P < 0.001] in addition to age (HR = 1.02, P = 0.004), ejection fraction (HR = 0.98, P = 0.004), and NYHA > II (HR = 1.86, P < 0.001) were predictors of death at 24 months. When the analysis was repeated dependent on NYHA-stage, MR-proADM (HR = 2.12, P < 0.001) and LVEF (HR = 0.96, P = 0.006) were significant markers, but only in patients with mild/moderate CHF. CONCLUSION: Our data suggest that MR proADM may be an important prognostic humoral marker, especially in mild/moderately symptomatic and non-ischaemic CHF patients. PMID- 19190024 TI - What is needed for a good 'Decaf'? PMID- 19190025 TI - St Jude Epic heart valve bioprostheses versus native human and porcine aortic valves - comparison of mechanical properties. AB - OBJECTIVES: The major problem with heart valve bioprostheses made from chemically treated porcine aortic valves is their limited longevity caused by gradual deterioration, which has a causal link with valve tissue mechanical properties. To our best knowledge, there are no published studies on the mechanical properties of modern, commercially available bioprostheses comparing them to native human valves. The objective of this study is to determine the mechanical properties of St Jude Epic bioprostheses and to compare them with native human and porcine aortic valves. METHODS: Leaflets from eight porcine aortic valves and six Epic bioprostheses were analyzed using uni-axial tensile tests in radial and circumferential directions. Mechanical properties of human valves have been previously published by our group. Results are represented as mean values+/-S.D. RESULTS: Circumferential direction. Modulus of elasticity of Epic bioprostheses in circumferential direction at the level of stress 1.0 MPa is 101.99+/-58.24 MPa, 42.3+/-4.96 MPa for native porcine and 15.34+/-3.84 MPa for human aortic valves. Ultimate stress is highest for Epic bioprostheses 5.77+/-1.94 MPa, human valves have ultimate stress of 1.74+/-0.29 MPa and porcine 1.58+/-0.26 MPa. Ultimate strain in circumferential direction is highest for human valves 18.35+/ 7.61% followed by 7.26+/-0.69% for porcine valves and 5.95+/-1.54% for Epic bioprostheses. Radial direction. Modulus of elasticity in radial direction is 9.18+/-1.81 MPa for Epic bioprostheses, 5.33+/-0.61 MPa for native porcine, and 1.98+/-0.15 MPa for human aortic valve leaflets. In the radial direction ultimate stress is highest for Epic bioprostheses 0.7+/-0.21 MPa followed by native porcine valves 0.55+/-0.11 MPa and 0.32+/-0.04 MPa for human valves. For human valves ultimate strain is 23.92+/-4.87%, for native porcine valves 8.57+/-0.8% and 7.92+/-1.74% for Epic bioprostheses. CONCLUSIONS: Epic bioprostheses have non linear stress-strain behavior similar to native valve tissue, but they are significantly stiffer and hence less elastic compared to native porcine and human aortic valves. These differences in mechanical properties may cause variations in stress distribution within leaflets of the bioprosthetic valves and accelerate their deterioration. PMID- 19190026 TI - Structural and functional changes of sulfated glycosaminoglycans in Xenopus laevis during embryogenesis. AB - Xenopus laevis is an excellent animal for analyzing early vertebrate development. Various effects of glycosaminoglycans (GAGs) on growth factor-related cellular events during embryogenesis have been demonstrated in Xenopus. To elucidate the relationship between alterations in fine structure and changes in the specificity of growth factor binding during Xenopus development, heparan sulfate (HS) and chondroitin/dermatan sulfate (CS/DS) chains were isolated at four different embryonic stages and their structure and growth factor-binding capacities were compared. The total amounts of both HS and CS/DS chains decreased from the pre midblastula transition to the gastrula stage, but increased exponentially during the following developmental stages. The length of HS chains was not significantly affected by development, whereas that of CS/DS chains increased with development. The disaccharide composition of GAGs in embryos also changed during development. The degree of sulfation of the HS chains gradually decreased with development. The predominant sulfation position in the CS/DS chains shifted from C4 to C6 of GalNAc during embryogenesis. Growth factor-binding experiments using a BIAcore system demonstrated that GAGs bound growth factors including fibroblast growth factors-1 and -2, midkine, and pleiotrophin, with comparable affinities. These affinities significantly varied during development, although the correlation between the structural alterations of GAGs and the change in the ability to bind growth factors remains to be clarified. The expression of saccharide sequences, which specifically interact with a growth factor, might be regulated during development. PMID- 19190027 TI - Receptor of advanced glycation end products (RAGE) positively regulates CD36 expression and reactive oxygen species production in human monocytes in diabetes. AB - INTRODUCTION: Advanced glycation end products (AGEs) engagement of a monocyte surface receptor (RAGE) induces atherosclerosis. AGEs also act as CD36 ligands. We studied reactive oxygen species (ROS) and CD36 expression after siRNA inhibition of RAGE expression in human monocytes. METHODS: We isolated monocytes from: a) 10 type 2 diabetics, and b) 5 age- and sex-matched healthy individuals. CD36 expression and ROS production were evaluated before and after RAGE knockdown. RESULTS: After incubation of monocytes with AGE + bovine serum albumin (BSA), CD36 expression and intracellular ROS increased significantly in all groups. In RAGE-knockdown monocytes, AGE-induced CD36 expression and ROS generation were also significantly inhibited. CONCLUSIONS: Blocking RAGE expression using siRNA in human monocytes led to a significant inhibition of CD36 expression and ROS production, suggesting a positive interaction between RAGE, CD36 expression and ROS generation in monocytes. PMID- 19190028 TI - The impact of novel treatment modalities on the therapeutic approach of cardiovascular diseases. PMID- 19190029 TI - Is high-sensitivity C-reactive protein associated with subclinical peripheral atherosclerosis? PMID- 19190030 TI - Renal artery dissection following marathon running. AB - A 38-year-old, previously healthy man presented with flank pain after competing in a marathon. Initial laboratory tests and urinalysis were essentially normal. Both contrast enhanced-computed tomography and magnetic resonance angiography showed an infarcted region of the left lower kidney without renal artery dissection. Thromboembolism was suspected, but further testing was negative. The diagnosis of renal artery dissection was established by angiogram, showing dissection of the segmental branch. The patient remained normotensive, maintained normal renal function, and had resolution of pain symptoms prior to discharge. On the basis of our experience and review of the literature, renal artery dissection occurs in otherwise healthy men and often goes undiagnosed. The management strategy tends to be conservative unless the patient develops progressive decline in renal function or worsening hypertension, with an excellent prognosis. This case also shows the importance of discussing the pros and cons of extreme physical exertion with all patients. PMID- 19190031 TI - Long-term exercise training selectively alters serum cytokines involved in fever. AB - Long-term exercise training selectively alters serum cytokines involved in fever. Chronic exercise training has a number of effects on the immune system that may mimic the physiological response to fever. Female rats that voluntarily exercise on running wheels develop an elevated daytime core temperature after several weeks of training. It remains to be seen whether the elevation in daytime temperature involves inflammatory patterns characteristic of an infectious fever. We assessed whether chronic exercise training in the rat would alter levels of cytokines involved in fever. Female Sprague Dawley rats at 45 days of age weighing 90-110 g were divided into two groups (exercise and sedentary) and housed at an ambient temperature of 22( degrees )C. Interleukin-1 beta (IL 1beta), interleukin-6 (IL-6), interleukin-10 (IL-10), tumor necrosis factor alpha (TNF-alpha), iron, and zinc levels were analyzed. Rats underwent 8 weeks of exercise on running wheels. Exercise led to altered levels of some key cytokines that are involved in fever. Exercise animals had significantly higher IL-1beta levels and lower IL-10 levels compared to sedentary animals. Although IL-6 levels were slightly lower in the exercise animals, these levels were not significantly affected by training. TNF-alpha activity was similar in the two groups. Training also led to a slight increase in serum zinc and decrease in serum unsaturated iron binding capacity (UIBC). The data suggest that chronic exercise training evokes immune responses that mimic some, but not all, aspects of fever. This may explain why exercise leads to elevated daytime core temperature. PMID- 19190032 TI - The role of impaired mitochondrial lipid oxidation in obesity. AB - Obesity represents a disruption in balancing fuel intake with energy expenditure in favor of energy conservation. Adiposity is known to be carefully regulated and, over time, highly resistant to major changes, raising questions about how energy homeostasis can become dysregulated in favor of fat accumulation. In obesity, the excess lipid accumulation represents a surfeit of energy, but those who are obese often experience rapid fatigue and decreased physical endurance, reflecting an energy deficiency. To develop an explanation for this apparent contradiction in energy homeostasis and the chronic overeating relative to energy used in obesity, a review of the literature was conducted. The resulting model of obesity is based on a growing body of research demonstrating that altered mitochondrial energy production, particularly in skeletal muscles, is a major anomaly capable of setting off a chain of metabolic events leading to obesity. Alterations in skeletal muscle mitochondria distribution and their oxidative and glycolytic energy capacities in obesity are described. The metabolic responses of obese and normal individuals to exercise are contrasted, and the effects of weight loss on energy production are presented. The effect of altered fat oxidation is considered in relation to energy regulation by the central nervous system and the development of major obesity comorbidities, including systemic inflammation, insulin resistance and diabetes, and cardiovascular disease. Recommendations for clinical intervention and additional research are proposed based on the model presented of impaired mitochondrial function in obesity. PMID- 19190033 TI - Effect of Withaferin A on the development and decay of thermotolerance in B16F1 melanoma: a preliminary study. AB - Protein synthesis inhibitors can suppress the development of thermotolerance in tumor tissues during [corrected] repeated heating. Withaferin A (WA), isolated from Withania somnifera has cytotoxic and inhibitory action on protein synthesis. In the present investigation, effect of WA on development and decay of thermotolerance in B16F1 melanoma was studied in C57BL mice. Tumors of 100 +/- 10 [corrected] mm(3) size were subjected to repeated hyperthermia (HT) at 43 degrees C for 30 minutes. WA was injected after first hyperthermia treatment. The tumor response was assessed by calculating the tumor growth delay (GD). The GD increased with increase in time gap between two hyperthermia treatments and was significantly higher (p < 0.05 to p < 0.001) in WA treated groups at all the respective time gaps (except at 0h and 120h) compared to hyperthermia alone group. WA increases the tumor response during repeated hyperthermia by reducing the magnitude of thermotolerance developed and by decreasing the recovery time from thermotolerance. PMID- 19190034 TI - Effects of a yoga program on cortisol rhythm and mood states in early breast cancer patients undergoing adjuvant radiotherapy: a randomized controlled trial. AB - Objectives. This study compares the effects of an integrated yoga program with brief supportive therapy in breast cancer outpatients undergoing adjuvant radiotherapy at a cancer center. METHODS: Eighty-eight stage II and III breast cancer outpatients are randomly assigned to receive yoga (n = 44) or brief supportive therapy (n = 44) prior to radiotherapy treatment. Assessments include diurnal salivary cortisol levels 3 days before and after radiotherapy and self ratings of anxiety, depression, and stress collected before and after 6 weeks of radiotherapy. RESULTS: Analysis of covariance reveals significant decreases in anxiety (P < .001), depression (P = .002), perceived stress (P < .001), 6 a.m. salivary cortisol (P = .009), and pooled mean cortisol (P = .03) in the yoga group compared with controls. There is a significant positive correlation between morning salivary cortisol level and anxiety and depression. CONCLUSION: Yoga might have a role in managing self-reported psychological distress and modulating circadian patterns of stress hormones in early breast cancer patients undergoing adjuvant radiotherapy. PMID- 19190035 TI - A descriptive analysis of health care coverage and concerns in West Central Wisconsin. AB - This study examines the level of health care coverage and health care concerns among a sample of 222 West Central Wisconsin residents. More than 68% of respondents felt that Wisconsin health care has at least major problems, and only 2.7% felt that there were no problems. Approximately 60% were extremely or very worried about not being able to afford health care services, whereas more than 70% were extremely or very worried that insurance companies care more about profits than patient care. These concerns translate into a desire for reform, with more than 90% of respondents saying it is extremely or very important for Wisconsin to make health care more affordable and accessible for all residents. Despite some differences between the insured and uninsured, the call for action to expand access and control costs was significant among both groups. These results can help guide the current policy debate on health care reform. PMID- 19190036 TI - Update on venous procedures performed in the office setting. AB - Treatment for chronic venous disease has evolved from hospital-based surgical procedures to minimally invasive office-based office procedures that provide shorter recovery periods, less postprocedural discomfort, and quicker return to normal activities. A specialty venous clinic, separate from the arterial practice, with a specialized professional health care team, provides a comfortable setting in which patients can have access to the most up-to-date treatment options. Sclerotherapy treatment for telangectasias, reticular veins, tributary varicosities, insufficient truncal veins, and incompetent perforating veins is a common therapy that is well suited for the office setting. Most office based minimally invasive venous procedures require little or no sedation. Tumescent anesthesia is safe and effective for ambulatory phlebectomy and endovenous ablation procedures. A calm, inviting atmosphere, confident and professional health care team, and prompt access to care will improve patient satisfaction and result in a successful, growing venous practice. PMID- 19190037 TI - Are children born to light drinkers not at high risk of developing clinically relevant cognitive-behavioural problems? A response to Kelly et al. PMID- 19190038 TI - Retroperitoneal repair of abdominal aortic aneurysm reduces bowel dysfunction. AB - OBJECTIVE: To assess the effect of intestinal manipulation and mesenteric traction on gastro-intestinal function and postoperative recovery in patients undergoing abdominal aortic aneurysm (AAA) repair. METHODS: Thirty-five patients undergoing AAA repair were randomised into 3 groups. Group I (n = 11) had repair via retroperitoneal approach while Group II (n = 12) and Group III (n = 12) were repaired via transperitoneal approach with bowel packed within the peritoneal cavity or exteriorised in a bowel bag respectively. Gastric emptying was measured pre-operatively (day 0), day 1 and day 3 using paracetamol absorption test (PAT) and area under curve (P(AUC)) was calculated. Intestinal permeability was measured using the Lactulose-Mannitol test. RESULTS: Aneurysm size, operation time and PAT (on day 0 and day 3) were similar in the three groups. On day 1, the P(AUC) was significantly higher in Group I, when compared with Group II and Group III (P = .02). Resumption of diet was also significantly earlier in Group I as compared to Group II and Group III. The intestinal permeability was significantly increased in Group II and Group III at day 1 when compared with day 0, with no significant increase in Group I. Retroperitoneal repair was also associated with significantly shorter intensive care unit (P = .04) and hospital stay (P = .047), when compared with the combined transperitoneal repair group (Group II and III). CONCLUSION: Retroperitoneal AAA repair minimises intestinal dysfunction and may lead to quicker patient recovery when compared to transperitoneal repair. PMID- 19190039 TI - How optimal is the medical management of patients prior to major reconstructive vascular surgery? The results of a cross-sectional study. AB - BACKGROUND: Risk factor modification is important in patients with vascular disease. Guidelines suggest that this patient population benefits from a medical regimen of antiplatelets, statins, beta-blockers, and angiotensin-converting enzyme (ACE) inhibitors. MATERIALS AND METHODS: The medical regimen of consecutive patients who presented for major vascular surgery intervention over 18 months was examined. RESULTS: Of the 325 patients identified, 176 (54%) were on antiplatelet treatment, 197 (61%) were on a statin, 180 (55%) on beta blockers, and 146 (54%) on ACE inhibitors. A high-risk subset of 94 smokers with known coronary artery disease, hypertension, and hyperlipidemia or diabetes was identified. In this subset, patients were more likely to be on pharmacologic risk factor modification. CONCLUSIONS: Despite recommendations, the pharmacologic risk factor modification of patients prior to vascular surgery remains suboptimal. Increased awareness of the problem and active participation of the vascular specialist are essential to improve compliance with established guidelines. PMID- 19190040 TI - Case study: chronic femoropopliteal prosthetic graft infection with exposed graft. AB - One of the most feared complications following vascular reconstruction is infection due to the attendant risks of limb loss, sepsis, or death. The reported incidence of infection following infrainguinal prosthetic graft infection is 2.5% with associated mortality rates and amputation rates of 18% and 41%, respectively. There are several options in treating infected prosthetic infrainguinal bypass grafts. Some authors have advocated complete removal of the infected graft with concomitant in situ revascularization using autogenous tissue or extra-anatomic bypass using either autogenous or prosthetic material, depending upon the clinical circumstances. Other authors have advocated attempting graft preservation to decrease the risk of amputation. Infected, thrombosed grafts are generally treated with graft excision alone with care taken to preserve collateral flow. The treatment options may also be influenced by the type of infection, as infections caused by gram-negative bacteria are thought to be more virulent than those associated with gram-positive bacteria. We recently treated a patient with an 18-month history of an exposed prosthetic graft in the groin, which was infected by Proteus mirabilis. Despite the extended period of graft exposure and despite gram-negative bacteria being the causative organism, the patient reported only intermittent drainage of pus from the groin. The management of this unusual infection forms the basis of this report. PMID- 19190041 TI - Ischemia-reperfusion injury in the lung: quantitation using electron microscopy. AB - BACKGROUND: The primary objectives of this study were to determine the time course of ischemia-reperfusion injury in an isolated rabbit lung model and to quantify this damage using electron microscopic methodology coupled with statistical analyses. MATERIALS AND METHODS: Eight groups of isolated rabbit lungs (n = 5 per group) were subjected to predetermined periods of ischemia reperfusion. Two hours of ischemia and 4 hours of reperfusion were concluded to be necessary to induce optimal ischemia-reperfusion injury in this model. Four other groups were subjected to 2 hours of ischemia followed by selected periods of reperfusion. These groups were compared to 4 control groups that were perfused for comparable time periods but without the initial ischemia. New quantitative methods were developed based on the average surface area of the alveoli and average number of alveoli per unit surface area, using scanning electron microscopic examination. RESULTS: Ischemia per se caused substantial damage. Restoration of volume and nutrients reversed this damage at 1 hour of reperfusion, but severe damage was evident at 4 hours of reperfusion, as reported by subjective and blinded examination. By using the new quantitative methods, there was a significant difference between the groups (P < .005) according to the time of post-ischemia-reperfusion, which correlated with the subjective evaluation of damage. CONCLUSIONS: These 2 new quantitative techniques provide an objective assessment of damage in the isolated rabbit lung model, suggesting that they warrant further consideration in similar studies of ischemia reperfusion injury. PMID- 19190043 TI - Quality of elective surgery in treatment centres. PMID- 19190042 TI - Crossmodal integration of object features: voxel-based correlations in brain damaged patients. AB - How does the brain bind together the different sensory features of objects to form meaningful, multimodal object representations? Human functional imaging findings implicate the left posterior superior temporal sulcus/middle temporal gyrus (pSTS/MTG) in crossmodal integration, while animal ablation findings support a hierarchical object processing model in which outputs from each sensory stream are integrated in perirhinal cortex (PRc) of the anteromedial temporal lobe. To determine which neural regions are necessary for integrating audiovisual object features, and which regions are necessary for understanding the meaning of crossmodal objects, we administered crossmodal (audio-visual) and unimodal (auditory, visual) integration tasks to 16 brain-damaged patients. We correlated patients' behavioural performance with measures of neural integrity (signal intensity) of each voxel across the brains of each patient. The integrity of bilateral anteromedial and temporopolar regions, but not pSTS/MTG, was significantly correlated with poorer crossmodal compared with unimodal integration performance, and with meaningful aspects of crossmodal integration. Additional analyses confirmed the negative crossmodal integration findings in the pSTS/MTG: performance on a sentence-picture matching control task was significantly correlated with MTG/STG voxel signal intensities, suggesting that a truncated range of signals in this region could not have been responsible for the lack of a significant correlation between integrity and crossmodal integration performance, and individual analyses of three patients with lesions in pSTS/MTG but spared anteromedial temporal cortex revealed equivalent unimodal and crossmodal integration performance. These results extend findings from the non human primate literature into the human domain by demonstrating that anteromedial temporal cortex is critically involved in crossmodal integration of object features. However, pSTS/MTG appears to play a supportive but non-essential role during crossmodal integration. Taken together, the present findings are consistent with a neurocognitive account of object representations which claims that anteromedial temporal lobe is critically involved in the formation and processing of complex, multimodal object representations. PMID- 19190044 TI - The management of patients with painful total knee replacement. AB - The management of patients with a painful total knee replacement requires careful assessment and a stepwise approach in order to diagnose the underlying pathology accurately. The management should include a multidisciplinary approach to the patient's pain as well as addressing the underlying aetiology. Pain should be treated with appropriate analgesia, according to the analgesic ladder of the World Health Organisation. Special measures should be taken to identify and to treat any neuropathic pain. There are a number of intrinsic and extrinsic causes of a painful knee replacement which should be identified and treated early. Patients with unexplained pain and without any recognised pathology should be treated conservatively since they may improve over a period of time and rarely do so after a revision operation. PMID- 19190045 TI - Medical negligence in orthopaedic surgery: a review of 130 consecutive medical negligence reports. AB - Payments by the NHS Litigation Authority continue to rise each year, and reflect an increase in successful claims for negligence against NHS Trusts. Information about the reasons for which Trusts are sued in the field of trauma and orthopaedic surgery is scarce. We analysed 130 consecutive cases of alleged clinical negligence in which the senior author had been requested to act as an expert witness between 2004 and 2006, and received information on the outcome of 97 concluded cases from the relevant solicitors. None of the 97 cases proceeded to a court hearing. Overall, 55% of cases were abandoned by the claimants' solicitors, and the remaining 45% were settled out of court. The cases were settled for sums ranging from pound 4500 to pound 2.7 million, the median settlement being pound 45,000. The cases that were settled out of court were usually the result of delay in treatment or diagnosis, or because of substandard surgical technique. PMID- 19190046 TI - Variables determining outcome in total hip replacement surgery. AB - In this study we hypothesised that anxiety/depression, one of five dimensions in the health-related quality of life (HRQoL) measurement tool EQ-5D, could predict outcome after total hip replacement surgery. Pre-operative and one-year post operative data from the Swedish Hip Arthroplasty Register, including 6158 patients with primary osteoarthritis of the hip, were analysed. In order to examine the association between anxiety and outcome with respect to pain and satisfaction an analysis of covariance was used. The pre-operative EQ-5D anxiety/depression dimension was a strong predictor for pain relief and patient satisfaction (p < 0.001). Orthopaedic surgeons involved in the care of patients eligible for total hip replacement surgery should be aware that mental health may influence post-operative pain and HRQoL. An appropriate assessment of mental health may enable a modification in the way these patients are managed in order to optimise the outcome after joint replacement surgery. PMID- 19190047 TI - Predictors of progression of osteoarthritis in femoroacetabular impingement: a radiological study with a minimum of ten years follow-up. AB - Although the association between femoroacetabular impingement and osteoarthritis is established, it is not yet clear which hips have the greatest likelihood to progress rapidly to end-stage disease. We investigated the effect of several radiological parameters, each indicative of a structural aspect of the hip joint, on the progression of osteoarthritis. Pairs of plain anteroposterior pelvic radiographs, taken at least ten years apart, of 43 patients (43 hips) with a pistol-grip deformity of the femur and mild (Tonnis grade 1) or moderate (Tonnis grade 2) osteoarthritis were reviewed. Of the 43 hips, 28 showed evidence of progression of osteoarthritis. There was no significant difference in the prevalence of progression between hips with initial Tonnis grade 1 or grade 2 osteoarthritis (p = 0.31). Comparison of the hips with and without progression of arthritis revealed a significant difference in the mean medial proximal femoral angle (81 degrees vs 87 degrees, p = 0.004) and the presence of the posterior wall sign (39% vs 7%, p = 0.02) only. A logistic regression model was constructed to predict the influence of these two variables in the development of osteoarthritis. Mild to moderate osteoarthritis in hips with a pistol-grip deformity will not progress rapidly in all patients. In one-third, progression will take more than ten years to manifest, if ever. The individual geometry of the proximal femur and acetabulum partly influences this phenomenon. A hip with cam impingement is not always destined for end-stage arthritic degeneration. PMID- 19190048 TI - The use of the cemented Exeter Universal femoral stem in a District General Hospital: a minimum ten-year follow-up. AB - We describe the survivorship of the Exeter femoral component in a District General Hospital. Between 1994 and 1996, 230 Exeter Universal cemented femoral components were implanted in 215 patients who were reviewed at a mean of 11.2 years (10 to 13). We used one acetabular implant, the Elite Ogee component, in 218 of the 230 hips. During the period of this study 76 patients (79 hips) died. Of the remaining 139 patients (151 hips), 121 were able to attend for radiological analysis at a minimum of ten years. One patient was lost to follow up. No femoral component was revised for aseptic loosening. Three hips were revised for deep infection and six acetabular components required revision, four for loosening and two for recurrent dislocation. Taking the 'worst-case scenario' including the one patient lost to follow-up, the overall survival rate was 94.4% at 13 years. Our results confirm excellent medium-term results for the Exeter Universal femoral component, implanted in a general setting. The excellent survival of this femoral component, when used in combination with the Ogee acetabular component, suggests that this is a successful pairing. PMID- 19190049 TI - Six-year results of a prospective study of metal ion levels in young patients with metal-on-metal hip resurfacings. AB - We describe the findings at six years in an ongoing prospective clinicoradiological and metal ion study in a cohort of 26 consecutive male patients with unilateral Birmingham Hip Resurfacing arthroplasties with one of two femoral head sizes (50 mm and 54 mm). Their mean age was 52.9 years (29 to 67). We have previously shown an early increase in the 24-hour urinary excretion of metal ions, reaching a peak at six months (cobalt) and one year (chromium) after operation. Subsequently there is a decreasing trend in excretion of both cobalt and chromium. The levels of cobalt and chromium in whole blood also show a significant increase at one year, followed by a decreasing trend until the sixth year. PMID- 19190050 TI - Arthrodesis of the knee using cannulated screws. AB - We retrospectively evaluated eight patients who underwent arthrodesis of the knee using cannulated screws. There were six women and two men, with a mean age of 53 years. The indications for arthrodesis were failed total knee arthroplasty, septic arthritis, tuberculosis, and recurrent persistent infection. Solid union was achieved in all patients at a mean of 6.1 months. One patient required autogenous bone graft for delayed union. One suffered skin necrosis which was treated with skin grafting. The mean limb-length discrepancy was 3.1 cm. On a visual analogue scale, the mean pain score improved from 7.9 to 3.3. According to the Knee Injury and Osteoarthritis Outcome score quality of life items, the mean score improved from 38.3 pre-operatively to 76.6 at follow-up. Cannulated screws provide a high rate of union in arthrodesis of the knee with minimal complications, patient convenience, and a simple surgical technique. PMID- 19190051 TI - Cementless Oxford unicompartmental knee replacement shows reduced radiolucency at one year. AB - We randomised 62 knees to receive either cemented or cementless versions of the Oxford unicompartmental knee replacement. The implants used in both arms of the study were similar, except that the cementless components were coated with porous titanium and hydroxyapatite. The tibial interfaces were studied with fluoroscopically-aligned radiographs. At one year there was no difference in clinical outcome between the two groups. Narrow radiolucent lines were seen at the bone-implant interfaces in 75% of cemented tibial components. These were partial in 43%, and complete in 32%. In the cementless implants, partial radiolucencies were seen in 7% and complete radiolucencies in none. These differences are statistically significant (p < 0.0001) and imply satisfactory bone ingrowth into the cementless implants. PMID- 19190052 TI - Meniscal root injury and spontaneous osteonecrosis of the knee: an observation. AB - We describe injuries to the posterior root of the medial meniscus in patients with spontaneous osteonecrosis of the medial compartment of the knee. We identified 30 consecutive patients with spontaneous osteonecrosis of the medial femoral condyle. The radiographs and MR imaging were reviewed. We found tears of the posterior root of the medial meniscus in 24 patients (80%). Of these, 15 were complete and nine were partial. Complete tears were associated with > 3 mm of meniscal extrusion. Neither the presence of a root tear nor the volume of the osteonecrotic lesion were associated with age, body mass index (BMI), gender, side affected, or knee alignment. The grade of osteoarthritis was associated with BMI. Although tears of the posterior root of the medial meniscus were frequently present in patients with spontaneous osteonecrosis of the knee, this does not prove cause and effect. Further study is warranted. PMID- 19190053 TI - Prevalence and characteristics of asymptomatic tears of the rotator cuff: an ultrasonographic and clinical study. AB - We undertook clinical and ultrasonographic examination of the shoulders of 420 asymptomatic volunteers aged between 50 and 79 years. MRI was performed in selected cases. Full-thickness tears of the rotator cuff were detected in 32 subjects (7.6%). The prevalence increased with age as follows: 50 to 59 years, 2.1%; 60 to 69 years, 5.7%; and 70 to 79 years, 15%. The mean size of the tear was less than 3 cm and tear localisation was limited to the supraspinatus tendon in most cases (78%). The strength of flexion was reduced significantly in the group with tears (p = 0.01). Asymptomatic tears of the rotator cuff should be regarded as part of the normal ageing process in the elderly but may be less common than hitherto believed. PMID- 19190054 TI - Large and massive tears of the rotator cuff: functional outcome and integrity of the repair after a mini-open procedure. AB - The aim of this study was to determine the functional outcome and rate of re tears following mini-open repair of symptomatic large and massive tears of the rotator cuff using a two-row technique. The 24 patients included in the study were assessed prospectively before and at a mean of 27 months (18 to 53) after surgery using the Constant and the Oxford Shoulder scores. Ultrasound examination was carried out at follow-up to determine the integrity of the repair. Patient satisfaction was assessed using a simple questionnaire. The mean Constant score improved significantly from 36 before to 68 after operation (p < 0.0001) and the mean Oxford Shoulder score from 39 to 20 (p < 0.0001). Four of the 24 patients (17%) had a re-tear diagnosed by ultrasound. A total of 21 patients (87.5%) were satisfied with the outcome of their surgery. The repair remained intact in 20 patients (83%). However, the small number of re-tears (four patients) in the study did not allow sufficient analysis to show a difference in outcome in relation to the integrity of the repair. PMID- 19190055 TI - Conservative treatment of lumbar spondylolysis in childhood and adolescence: the radiological signs which predict healing. AB - It has been noted that bony union of a pars defect can be achieved in children if they wear a trunk brace. Our aim was to evaluate how the stage of the defect on CT and the presence or absence of high signal change in the adjacent pedicle on T2-weighted MRI were related to bony healing. We treated 23 children conservatively for at least three months. There were 19 boys and four girls with a mean age of 13.5 years (7 to 17). They were asked to refrain from sporting activity and to wear a Damen soft thoracolumbosacral type brace. There were 41 pars defects in 23 patients. These were classified as an early, progressive or terminal stage on CT. The early-stage lesions had a hairline crack in the pars interarticularis, which became a gap in the progressive stage. A terminal-stage defect was equivalent to a pseudarthrosis. On the T2-weighted MR scan the presence or absence of high signal change in the adjacent pedicle was assessed and on this basis the defects were divided into high signal change-positive or negative. Healing of the defect was assessed by CT. In all, 13 (87%) of the 15 early defects healed. Of 19 progressive defects, only six (32%) healed. None of the seven terminal defects healed. Of the 26 high signal change-positive defects 20 (77%) healed after conservative treatment whereas none of the high signal change-negative defects did so. We concluded that an early-stage defect on CT and high signal change in the adjacent pedicle on a T2-weighted MR scan are useful predictors of bony healing of a pars defect in children after conservative treatment. PMID- 19190056 TI - Health-related quality of life (EQ-5D) before and one year after surgery for lumbar spinal stenosis. AB - We investigated the pre-operative and one-year post-operative health-related quality of life (HRQoL) outcome by using a Euroqol (EQ-5D) questionnaire in 230 patients who underwent surgery for lumbar spinal stenosis. Data were obtained from the National Swedish Registry for operations on the lumbar spine between 2001 and 2002. We analysed the pre- and postoperative quality of life data, age, gender, smoking habits, pain and walking ability. The relative differences were compared to a Swedish EQ-5D population survey. The mean age of the patients was 66 years, and there were 123 females (53%). Before the operation 62 (27%) of the patients could walk more than 500 m. One year after the operation 150 (65%) were able to walk 500 m or more. The mean EQ-5D score improved from 0.36 to 0.64, and the HRQoL improved in 184 (80%) of the patients. However, they did not reach the level reported by a matched population sample (mean difference 0.18). Women had lower pre- and post-operative EQ-5D scores than men. Severe low back pain was a predictor for a poor outcome. PMID- 19190057 TI - Immediate primary skin closure in type-III A and B open fractures: results after a minimum of five years. AB - Between June 1999 and May 2003 we undertook direct primary closure of the skin wounds of 173 patients with Gustilo and Anderson grade-IIIA and grade-IIIB open fractures. These patients were selected from a consecutive group of 557 with type III injuries presenting during this time. Strict criteria for inclusion in the study included debridement within 12 hours of injury, no sewage or organic contamination, no skin loss either primarily or secondarily during debridement, a Ganga Hospital open injury skin score of 1 or 2 with a total score of ten or less, the presence of bleeding skin margins, the ability to approximate wound edges without tension and the absence of peripheral vascular disease. In addition, patients with polytrauma were excluded. At a mean follow-up of 6.2 years (5 to 7), the outcome was excellent in 150 (86.7%), good in 11 (6.4%) and poor in 12 (6.9%). A total of 33 complications occurred in 23 patients including superficial infection in 11, deep infection in five and the requirement for a secondary skin flap in three. Six patients developed nonunion requiring further surgery, one of whom declined additional measures to treat an established infected nonunion. Immediate skin closure when performed selectively with the above indications proved to be a safe procedure. PMID- 19190058 TI - Incidence of MRSA surgical-site infection in MRSA carriers in an orthopaedic trauma unit. AB - We examined the incidence of infection with methicillin-resistant Staphylococcus aureus (MRSA) in patients admitted to the Leicester Royal Infirmary Trauma Unit between January 2004 and June 2006. The influence of MRSA status at the time of their admission was examined, together with age, gender and diagnosis, using multi-variant analysis. Of 2473 patients, 79 (3.2%) were MRSA carriers at the time of admission and 2394 (96.8%) were MRSA-negative. Those carrying MRSA at the time of admission were more likely to develop surgical site infection with MRSA (7 of 79 patients, 8.8%) than non-MRSA carriers (54 of 2394 patients, 2.2%, p < 0.001). Further analysis showed that hip fracture and increasing age were also risk factors with a linear increase in relative risk of 1.8% per year. MRSA carriage at admission, age and the pathology are all associated with an increased rate of developing MRSA wound infection. Identification of such risk factors at admission helps to target health-care resources, such the use of glycopeptide antibiotics at induction and the 'building-in' of increased vigilance for wound infection pre-operatively. PMID- 19190059 TI - Review of Kinemax knee arthroplasty performed at the NHS Treatment Centre, Weston Super-Mare. AB - As part of the government's initiative to reduce waiting times for major joint surgery in Wales, the Cardiff and Vale NHS Trust sent 224 patients (258 knees) to the NHS Treatment Centre in Weston-Super-Mare for total knee replacement. The Kinemax total knee replacement system was used in all cases. The cumulative survival rate at three years was 79.2% (95% confidence interval (CI) 69.2 to 86.8) using re-operation for any cause as an endpoint and 85.3% (95% CI 75.9 to 91.8) using aseptic revision as an endpoint. This is significantly worse than that recorded in the published literature. These poor results have resulted in a significant impact on our service. PMID- 19190060 TI - Fibular centralisation for the reconstruction of defects of the tibial diaphysis and distal metaphysis after excision of bone tumours. AB - We evaluated the results of fibular centralisation as a stand alone technique to reconstruct defects that occurred after resection of tumours involving the tibial diaphysis and distal metaphysis. Between January 2003 and December 2006, 15 patients underwent excision of tumours of the tibial diaphysis or distal metaphysis and reconstruction by fibular centralisation. Their mean age was 17 years (7 to 40). Two patients were excluded; one died from the complications of chemotherapy and a second needed a below-knee amputation for a recurrent giant cell tumour. A total of 13 patients were reviewed after a mean follow-up of 29 months (16 to 48). Only 16 of 26 host graft junctions united primarily. Ten junctions in ten patients needed one or more further procedure before union was achieved. At final follow-up 12 of the 13 patients had fully united grafts; 11 walked without aids. The mean time to union at the junctions that united was 12 months (3 to 36). The mean Musculoskeletal Tumor Society Score was 24.7 (16 to 30). Fibular centralisation is a durable reconstruction for defects of the tibial diaphysis and distal metaphysis with an acceptable functional outcome. Stable osteosynthesis is the key to successful union. Additional bone grafting is recommended for patients who need postoperative radiotherapy. PMID- 19190061 TI - The effect of early surgical treatment on recovery in patients with metastatic compression of the spinal cord. AB - We analysed the influence of the timing of surgery (< 48 hours, group 1, 21 patients vs > 48 hours, group 2, 14 patients) on the neurological outcome and restoration of mobility in 35 incomplete tetra- and paraplegic patients with metastatic spinal-cord compression. Pain and neurological symptoms were assessed using the American Spinal Injury Association impairment scale. More improvement was found in group 1 than in group 2 when comparing the pre-operative findings with those both immediately post-operatively (p = 0.021) and those at follow-up at four to six weeks (p = 0.010). In group 1 the number of pre-operatively mobile patients increased from 17 (81%) to 19 patients (90%) whereas the number of mobile patients in group 2 changed from nine (64%) to ten (71%). These results suggest that early surgical treatment in patients with metastatic spinal-cord compression gives a better neurological outcome even in a palliative situation. PMID- 19190062 TI - The effectiveness of a programme for neonatal hip screening over a period of 40 years: a follow-up of the New Plymouth experience. AB - Since September 1964, neonates born in New Plymouth have undergone clinical examination for instability of the hip in a structured clinical screening programme. Of the 41 563 babies born during this period, 1639 were diagnosed as having unstable hips and 663 (1.6%) with persisting instability were splinted, five of which failed. Also, three unsplinted hips progressed to congenital dislocation, and there were four late-presenting (walking) cases, giving an overall failure rate of 0.29 per 1000 live births, with an incidence of late walking congenital dislocation of the hip of 0.1 per 1000 live births. This study confirms that clinical screening for neonatal instability of the hip by experienced orthopaedic examiners significantly reduces the incidence of late presenting (walking) congenital dislocation of the hip. PMID- 19190063 TI - Methicillin-resistant Staphylococcus aureus in orthopaedic trauma: identification of risk factors as a strategy for control of infection. AB - We have conducted a case-control study over a period of ten years comparing both deep infection with methicillin-resistant staphylococcus aureus (MRSA) and colonised cases with a control group. Risk factors associated with deep infection were vascular diseases, chronic obstructive pulmonary disease, admission to a high-dependency or an intensive-care unit and open wounds. Those for colonisation were institutional care, vascular diseases and dementia. Older age was a risk factor for any MRSA infection. The length of hospital stay was dramatically increased by deep infection. These risk factors are useful in identifying higher risk patients who may be more susceptible to MRSA infection. A strategy of early identification and isolation may help to control its spread in trauma units. PMID- 19190064 TI - Early palsy of the sciatic nerve due to heterotopic ossification after surgery for fracture of the posterior wall of the acetabulum. AB - We describe a patient in whom an initially intact sciatic nerve became rapidly encased in heterotopic bone formed in the abductor compartment after reconstruction of the posterior wall of the acetabulum following fracture. Prompt excision and neural release followed by irradiation and administration of indometacin resulted in a full neurological recovery and no recurrence 27 months later. PMID- 19190065 TI - The effect of parecoxib and indometacin on tendon-to-bone healing in a bone tunnel: an experimental study in rats. AB - Conventional non-steroidal anti-inflammatory drugs (NSAIDs) and newer specific cyclo-oxygenase-2 (cox-2) inhibitors are commonly used in musculoskeletal trauma and orthopaedic surgery to reduce the inflammatory response and pain. These drugs have been reported to impair bone metabolism. In reconstruction of the anterior cruciate ligament the hamstring tendons are mainly used as the graft of choice, and a prerequisite for good results is healing of the tendons in the bone tunnel. Many of these patients are routinely given NSAIDs or cox-2 inhibitors, although no studies have elucidated the effects of these drugs on tendon healing in the bone tunnel. In our study 60 female Wistar rats were randomly allocated into three groups of 20. One received parecoxib, one indometacin and one acted as a control. In all the rats the tendo-Achillis was released proximally from the calf muscles. It was then pulled through a drill hole in the distal tibia and sutured anteriorly. The rats were given parecoxib, indometacin or saline intraperitoneally twice daily for seven days. After 14 days the tendon/bone tunnel interface was subjected to mechanical testing. Significantly lower maximum pull-out strength (p < 0.001), energy absorption (p < 0.001) and stiffness (p = 0.035) were found in rats given parecoxib and indometacin compared with the control group, most pronounced with parecoxib. PMID- 19190066 TI - Osteogenic activity of human fracture haematoma-derived progenitor cells is stimulated by low-intensity pulsed ultrasound in vitro. AB - The haematoma occurring at the site of a fracture is known to play an important role in bone healing. We have recently shown the presence of progenitor cells in human fracture haematoma and demonstrated that they have the capacity for multilineage mesenchymal differentiation. There have been many studies which have shown that low-intensity pulsed ultrasound (LIPUS) stimulates the differentiation of a variety of cells, but none has investigated the effects of LIPUS on cells derived from human fracture tissue including human fracture haematoma-derived progenitor cells (HCs). In this in vitro study, we investigated the effects of LIPUS on the osteogenic activity of HCs. Alkaline phosphatase activity, osteocalcin secretion, the expression of osteoblast-related genes and the mineralisation of HCs were shown to be significantly higher when LIPUS had been applied but without a change in the proliferation of the HCs. These findings provide evidence in favour of the use of LIPUS in the treatment of fractures. PMID- 19190067 TI - Repair of defects and containment in revision total knee replacement: a comparative biomechanical analysis. AB - The treatment of bony defects of the tibia at the time of revision total knee replacement is controversial. The place of compacted morsellised bone graft is becoming established, particularly in contained defects. It has previously been shown that the initial stability of impaction-grafted trays in the contained defects is equivalent to that of an uncemented primary knee replacement. However, there is little biomechanical evidence on which to base a decision in the treatment of uncontained defects. We undertook a laboratory-based biomechanical study comparing three methods of graft containment in segmental medial tibial defects and compared them with the use of a modular metal augment to bypass the defect. Using resin models of the proximal tibia with medial defects representing either 46% or 65% of the medial cortical rim, repair of the defect was accomplished using mesh, cement or a novel bag technique, after which impaction bone grafting was used to fill the contained defects and a tibial component was cemented in place. As a control, a cemented tibial component with modular metal augments was used in identical defects. All specimens were submitted to cyclical mechanical loading, during which cyclical and permanent tray displacement were determined. The results showed satisfactory stability with all the techniques except the bone bag method. Using metal augments gave the highest initial stability, but obviously lacked any potential for bone restoration. PMID- 19190068 TI - Rembrandt's 'Beggar with a wooden leg' and other comparable prints. AB - Rembrandt's etching of a beggar with a wooden leg is notable because the two lower limbs of the presumed beggar are present and not deformed. Using the facilities of four specialised Dutch art institutes, we carried out a systematic investigation to find other etchings and engravings of subjects with artificial legs supporting non-amputated limbs, from the period 1500 to 1700 AD. We discovered 28 prints produced by at least 18 artists. Several offered clues to a disorder of a knee, the lower leg or the foot. All individuals were adult males, suggesting the probability of traumatic lesions. We conclude that in this period artificial legs were not only used in the case of absence of part of a lower limb, but also for other reasons, notably disorders of the knee, lower leg or foot. They may also have been used to attract compassion. PMID- 19190069 TI - The female-male disparity in obesity prevalence among black American young adults: contributions of sociodemographic characteristics of the childhood family. AB - BACKGROUND: In the United States, black women are at much greater risk of obesity than are black men. Little is known about the factors underlying this disparity. OBJECTIVE: We explored whether childhood sociodemographic factors (parental education, single-mother household, number of siblings, number of minors in household, birth order, and female caregiver's age) were associated with the gender disparity in obesity prevalence in young black adults in the United States. DESIGN: An analytic data set (n = 7747) was constructed from the nationally representative National Longitudinal Study of Adolescent Health. Childhood sociodemographic factors were assessed in 1994-1995 in nonimmigrant black and white youths aged 11-19 y. Obesity was assessed in 2001-2002. For each childhood sociodemographic factor, we evaluated whether the prevalence difference (female obesity minus male obesity) was modified by the factor. We described the contribution of each variable category to the overall prevalence difference. RESULTS: In unadjusted and multivariable-adjusted models, parental education consistently modified gender disparity in blacks (P = 0.01). The gender gap was largest with low parental education (16.7% of men compared with 45.4% of women were obese) and smallest with high parental education (28.5% of men compared with 31.4% of women were obese). In whites, there was little overall gender difference in obesity prevalence. CONCLUSIONS: To our knowledge, this was the first study to document that the gender disparity in obesity prevalence in young black adults is concentrated in families with low parental education. In these low-socioeconomic status families, obesity development is either under the control of distinct mechanisms in each gender, or men and women from these households adopt different obesity-related behaviors. PMID- 19190070 TI - Effect of 2 pieces of nutritional advice on folate status in Swedish women: a randomized controlled trial. AB - BACKGROUND: Ten years after the introduction of mandatory folic acid fortification in the United States, Canada, and Costa Rica, the issue is still under debate in several countries, and Sweden recently decided against mandatory fortification. OBJECTIVE: The objective was to determine the folate status of women after an intervention involving 2 Swedish dietary recommendations: a food recommendation (bread) and a complete meal recommendation (breakfast). DESIGN: Fifty-one free-living women with normal folate status participated in a 12-wk controlled intervention trial. Subjects were randomly assigned to one of the following interventions: apple juice (control group; n = 17), a breakfast providing 125 microg folate (breakfast group; n = 17), or 5 slices of whole-meal bread to be eaten over the course of the day, which provided 70 microg folate (bread group; n = 17). Folate status was assessed on the basis of concentrations of erythrocyte folate, serum folate, and plasma total homocysteine (tHcy) at baseline and at weeks 8 and 12 of the trial. RESULTS: In the breakfast group, initial median concentrations of erythrocyte folate (805 nmol/L) increased by 172 nmol/L (95% CI: 24, 293; P = 0.02) relative to the control. The relative increase in initial serum folate (2 nmol/L, 95% CI: 0, 5; P = 0.06) was nonsignificant. The initial tHcy concentration (8.7 micromol/L) decreased by 2.3 micromol/L (95% CI: -1, -3.4; P < 0.01). In the bread group, the initial tHcy concentration (9.1 micromol/L) decreased nonsignificantly by 1.4 micromol/L (95% CI: 0, -2.8; P = 0.08) relative to the control group, whereas other outcomes were stable. CONCLUSIONS: The folate status of the subjects improved after regular consumption of the breakfast meal. The additional folate intake from the bread maintained the folate status but was not sufficient to improve it. PMID- 19190071 TI - Leptin at 14 y of age: an ongoing story. AB - The cloning of the ob gene and its gene product leptin has led to the elucidation of a robust physiologic system that maintains constancy of fat stores. Leptin is a peptide hormone secreted by adipose tissue and regulates adipose tissue mass and energy balance. Recessive mutations in the leptin gene are associated with massive obesity in mice and in some humans, which establishes a genetic basis for obesity. Leptin circulates in blood and acts on the brain to regulate food intake and energy expenditure. When fat mass decreases, plasma leptin concentrations decrease, which stimulates appetite and suppresses energy expenditure until fat mass is restored. When fat mass increases, leptin concentrations increase, which suppresses appetite until weight is lost. This system maintains homeostatic control of adipose tissue mass. PMID- 19190072 TI - Estimates of excess deaths associated with body mass index and other anthropometric variables. AB - BACKGROUND: Estimates of excess mortality associated with body mass index (BMI; in kg/m(2)) have been calculated for the US population. OBJECTIVE: The objective of this article is to compare the excess mortality associated with BMI levels to the excess mortality associated with other anthropometric variables. DESIGN: For the 1988-1994 Third National Health and Nutrition Examination Survey, estimates of excess deaths were calculated for standard BMI levels and for comparable levels of percentage body fat, waist circumference, hip and arm circumferences, waist-hip ratio, the sum of 4 skinfold thicknesses, and waist-stature ratio. The outcome measure is the percentage of deaths in the full sample in excess of those predicted for the reference category. RESULTS: For the level equivalent to BMI <18.5, estimates of excess deaths ranged from 0.3% for waist-hip ratio to 2.4% for percentage body fat. All except waist circumference, waist-hip ratio, and waist-stature ratio were significantly greater than zero (P < 0.05). For the level equivalent to BMI 25 to <30, the percentage of excess deaths was 0.1% for percentage body fat and negative for all other variables; estimates were significantly below zero only for circumferences and waist-stature ratio. For the level equivalent to BMI > or = 30, estimates ranged from -1.7% for waist circumference to 1.5% for percentage of fat; none were significantly different from zero. Estimates for all-cause mortality, obesity-related causes of death, and other causes of death showed no statistically significant or systematic differences between BMI and other variables. CONCLUSION: In this population-based study, attributable fractions of deaths were similar across measures. PMID- 19190073 TI - Dietary sources of vitamin B-12 and their association with plasma vitamin B-12 concentrations in the general population: the Hordaland Homocysteine Study. AB - BACKGROUND: Limited information is available on the association between vitamin B 12 status and intake from different dietary sources. OBJECTIVE: We investigated the relation of dietary intake of different food items with plasma vitamin B-12 concentrations in the general population. DESIGN: A cross-sectional, population based study of 5937 subjects in 2 age groups (47-49 and 71-74 y) from the Hordaland Homocysteine Study in Norway was conducted by using a food-frequency questionnaire and measurements of plasma vitamin B-12 concentrations. RESULTS: A significant difference in plasma vitamin B-12 concentrations was observed with increasing total vitamin B-12 intake. A plateau was reached at an intake of approximately 10 microg/d. Plasma vitamin B-12 was associated with intakes of increasing amounts of vitamin B-12 from dairy products or fish (P for trend <0.001 for both) but not with intakes of vitamin B-12 from meat or eggs. For the same content of vitamin B-12, intake from dairy products led to the greatest increase in plasma vitamin B-12. Total intake of vitamin B-12, particularly from milk and fish, decreased the risk of vitamin B-12 concentrations <200 pmol/L and impaired vitamin B-12 function (vitamin B-12 <200 pmol/L and methylmalonic acid >0.27 micromol/L) in the total group and in 71-74-y-old subjects. CONCLUSIONS: Dietary intake of dairy products and fish are significant contributors to plasma vitamin B-12 and may improve plasma vitamin B-12 status. Vitamin B-12 appears to be more bioavailable from dairy products; guidelines for improving vitamin B-12 status should take this into consideration. PMID- 19190074 TI - Retrospective exposure assessment for carcinogenic agents in bitumen waterproofing industry in Finland and denmark. AB - OBJECTIVES: The purpose of the study was (i) to identify the carcinogenic agents that may cause confounding when studying the exposure-response relationship between bitumen fume exposure and cancer among roofing membrane-manufacturing workers and roofers and (ii) to assess exposures to the identified carcinogens and bitumen fume in roofing membrane manufacturing and roofing in Finland and Denmark from 1950 to 2005. METHODS: Information on the use of carcinogenic agents and other relevant data were collected through semi-structured interviews of senior employees in the industry. Semi-quantitative exposure assessments were made on the basis of available measurement data and information obtained from the interviews and literature. RESULTS: Most of the production line workers in roofing membrane plants in Finland were exposed to asbestos until the mid-1970s. Also, some of the mixer operators in the plants were exposed to asbestos in Finland during the 1970s and in Denmark from the mid-1960s to the mid-1980s. In both countries, coal tar pitch was used in roofing membrane manufacturing until the mid-1960s, and consequently, exposure to polycyclic aromatic hydrocarbons (PAHs) in the plants was high in the 1950s and still significant in the early 1960s. Exposure of production line workers to quartz dust was high until the 1980s and is still relatively high compared with current occupational exposure limit values. Bitumen roofers' exposure to coal tar-derived PAHs may have been significant in both countries until the end of 1960s. Roofers' exposure to asbestos and quartz was estimated to have been near background level. CONCLUSIONS: The estimated average annual exposures to asbestos, coal tar-derived PAHs and quartz dust in the bitumen waterproofing industry in Finland and Denmark were significant in the past but have a clear declining trend. Exposure to bitumen fume was found to follow a similar trend. PMID- 19190075 TI - Clinical and pharmacological phase I evaluation of Exherin (ADH-1), a selective anti-N-cadherin peptide in patients with N-cadherin-expressing solid tumours. AB - BACKGROUND: Upregulation of N-cadherin promotes dysregulated cell growth, motility, invasiveness, plus maintenance of vascular stability and is associated with cancer progression in several human tumour types. N-cadherin is expressed also on tumour cells and the anti-N-cadherin cyclic pentapeptide ADH-1, tested in the present study, can exert a direct antitumour effect. PATIENTS AND METHODS: Adult patients with advanced solid malignancies expressing N-cadherin on tumour biopsies carried out in the previous 12 months received escalating i.v. doses of ADH-1 given weekly (initially for 3 of 4 weeks, then every week). Plasma pharmacokinetics (PK) was studied at cycle 1. Blood flow changes were assessed after first dosing in all patients treated in the initial regimen. RESULTS: In all, 129 patients were screened, 65 (50%) were N-cadherin positive, and 30 were enrolled. The doses ranged from 150 to 2400 mg/m(2); no maximum tolerated dose was reached. Treatment was well tolerated with asthenia as the most frequent adverse event. Two patients with ovarian cancer showed prolonged disease stabilisation while one patient with fallopian tube carcinoma achieved a mixed response. PK was linear in the range of doses tested. CONCLUSION: ADH-1 is the first anti-N-cadherin compound tested in humans. In N-cadherin-positive patients, ADH-1 showed an acceptable toxicity profile, linear PK and hints of antitumour activity in gynaecological cancers. PMID- 19190076 TI - Payments to normal healthy volunteers in phase 1 trials: avoiding undue influence while distributing fairly the burdens of research participation. AB - Clinical investigators must engage in just subject recruitment and selection and avoid unduly influencing research participation. There may be tension between the practice of keeping payments to participants low to avoid undue influence and the requirements of justice when recruiting normal healthy volunteers for phase 1 drug studies. By intentionally keeping payments low to avoid unduly influenced participation, investigators, on the recommendation or insistence of institutional review boards, may be targeting or systematically recruiting healthy adult members of lower socio-economic groups for participation in phase 1 studies. Investigators are at risk of routinely failing to fulfill the obligation of justice, which prohibits the systematic targeting and recruiting of subjects for reasons unrelated to the nature of the study. Insofar as we take seriously the obligation to engage in just subject recruitment and selection, I argue that we must acknowledge the implications low payments might have for subject recruitment and selection and examine the effect of low payments. If low payments de facto target the less well-off for phase 1 studies, we must defend the priority ranking of the obligation to avoid undue influence over the obligation of justice or adopt an alternative recruitment approach. This paper identifies a number of alternatives to the current system of low-value payments to research participants. PMID- 19190077 TI - Clinical aspects of SDHx-related pheochromocytoma and paraganglioma. AB - Paragangliomas (PGLs) derive from either sympathetic chromaffin tissue in adrenal and extra-adrenal abdominal or thoracic locations, or from parasympathetic tissue of the head and neck. Mutations of nuclear genes encoding subunits B, C, and D of the mitochondrial enzyme succinate dehydrogenase (SDHB 1p35-p36.1, SDHC 1q21, SDHD 11q23) give rise to hereditary PGL syndromes PGL4, PGL3, and PGL1 respectively. The susceptibility gene for PGL2 on 11q13.1 remains unidentified. Mitochondrial dysfunction due to SDHx mutations have been linked to tumorigenesis by upregulation of hypoxic and angiogenesis pathways, apoptosis resistance and developmental culling of neuronal precursor cells. SDHB-, SDHC-, and SDHD associated PGLs give rise to more or less distinct clinical phenotypes. SDHB mutations mainly predispose to extra-adrenal, and to a lesser extent, adrenal PGLs, with a high malignant potential, but also head and neck paragangliomas (HNPGL). SDHD mutations are typically associated with multifocal HNPGL and usually benign adrenal and extra-adrenal PGLs. SDHC mutations are a rare cause of mainly HNPGL. Most abdominal and thoracic SDHB-PGLs hypersecrete either norepinephrine or norepinephrine and dopamine. However, only some hypersecrete dopamine, are biochemically silent. The biochemical phenotype of SDHD-PGL has not been systematically studied. For the localization of PGL, several positron emission tomography (PET) tracers are available. Metastatic SDHB-PGL is the best localized by [(18)F]-fluorodeoxyglucose PET. The identification of SDHx mutations in patients with PGL is warranted for a tailor-made approach to the biochemical diagnosis, imaging, treatment, follow-up, and family screening. PMID- 19190078 TI - The MPA mouse breast cancer model: evidence for a role of progesterone receptors in breast cancer. AB - More than 60% of all breast neoplasias are ductal carcinomas expressing estrogen (ER) and progesterone receptors (PR). By contrast, most of the spontaneous, chemically or mouse mammary tumor virus induced tumors, as well as tumors arising in genetically modified mice do not express hormone receptors. We developed a model of breast cancer in which the administration of medroxyprogesterone acetate to BALB/c female mice induces mammary ductal carcinomas with a mean latency of 52 weeks and an incidence of about 80%. These tumors are hormone-dependent (HD), metastatic, express both ER and PR, and are maintained by syngeneic transplants. The model has been further refined to include mammary carcinomas that evolve through different stages of hormone dependence, as well as several hormone responsive cell lines. In this review, we describe the main features of this tumor model, highlighting the role of PR as a trigger of key signaling pathways mediating tumor growth. In addition, we discuss the relevance of this model in comparison with other presently used breast cancer models pointing out its advantages and limitations and how, this model may be suitable to unravel key questions in breast cancer. PMID- 19190079 TI - Mutational analysis of the BRAF, RAS and EGFR genes in human adrenocortical carcinomas. AB - The serine/threonine kinase B-Raf plays a key role in the Ras/Raf/MEK/ERK pathway that relays extracellular signals for cell proliferation and survival. Several types of human malignancies harbor activating BRAF mutations, most frequently a V600E substitution. The epidermal growth factor receptor (EGFR), a transmembrane tyrosine kinase (TK) receptor that mediates proliferation and survival signaling, is expressed in a wide variety of normal and neoplastic tissues. EGFR inhibitors have produced objective responses in patients with non-small cell lung carcinomas harboring activating EGFR TK domain somatic mutations. We evaluated the presence of mutations in BRAF (exons 11 and 15), KRAS (exons 1 and 2), NRAS (exons 1 and 2), and EGFR (exons 18-21) in adrenal carcinomas (35 tumor specimens and two cell lines) by DNA sequencing. BRAF mutations were found in two carcinomas (5.7%). Four carcinomas (11.4%) carried EGFR TK domain mutations. One specimen carried a KRAS mutation, and another carried two NRAS mutations. No mutations were found in the two adrenocortical cell lines. BRAF- and EGFR-mutant tumor specimens exhibited stronger immunostaining for the phosphorylated forms of the MEK and ERK kinases than their wild-type counterparts. EGFR-mutant carcinomas exhibited increased phosphorylation of EGFR (Tyr 992) compared with wild-type carcinomas. We conclude that BRAF, RAS, and EGFR mutations occur in a subset of human adrenocortical carcinomas. Inhibitors of the Ras/Raf/MEK/ERK and EGFR pathways represent candidate targeted therapies for future clinical trials in carefully selected patients with adrenocortical carcinomas harboring respective activating mutations. PMID- 19190080 TI - Resistance to chemotherapy and hormone therapy in endometrial cancer. AB - Endometrial cancer is the most common gynecological malignancy in developed countries and represents the eighth leading cause of cancer related death in women. The growing incidence of endometrial cancer leads scientists and oncologists to identify effective preventive measures and also molecular markers for diagnosis and prognosis. Chemotherapy and hormone therapy is the mainstay treatment option for advanced and recurrent endometrial cancer and response to therapy is one of the most important factor which favors prognosis and overall survival. In recent years, there have been major advances in the treatment of patients with endometrial cancer. Despite advances made in the treatment of this cancer, the overall survival of patients has not significantly improved because considerable number of patients harbor tumor refractory to these therapies and the majority of the initially responsive tumors become refractory to treatments. Therefore, determination of sensitivity/resistance is becoming increasingly important for individualization of endometrial cancer therapy. The aim of this review is to present the existing knowledge about the molecular markers that could play a crucial role in determining resistance to chemo- and hormone therapy. Extensive literature search for the cell signaling pathways and factors responsible for chemoresistance have been performed and reviewed. Several recent studies suggest that deregulations in the apoptotic pathways (such as p53, Fas/FasL, Bcl-2 family proteins, inhibitor of apoptosis proteins), survival pathways (PI3K/AKT, MAPK), hormone receptor signaling pathways (progesterone receptor), Cyclooxygenase-2 and Her-2 are considered as key factors involved in the onset and maintenance of therapeutic resistance, suggesting that resistance is a multi-factorial phenomenon. PMID- 19190081 TI - The National Pharmaceuticals Strategy: Rest in peace, revive or renew? PMID- 19190082 TI - Sex of the cell dictates its response: differential gene expression and sensitivity to cell death inducing stress in male and female cells. AB - Sexual dimorphisms are typically attributed to the hormonal differences arising once sex differentiation has occurred. However, in some sexually dimorphic diseases that differ in frequency but not severity, the differences cannot be logically connected to the sex hormones. Therefore, we asked whether any aspect of sexual dimorphism could be attributed to chromosomal rather than hormonal differences. Cells taken from mice at d 10.5 postconception (PC) before sexual differentiation, at d 17.5 PC after the first embryonic assertion of sexual hormones, and at postnatal day 17 (puberty) were cultured and exposed to 400 microM ethanol or 20 microM camptothecin or to infection with influenza A virus (multiplicity of infection of 5). The results showed that untreated male and female cells of the same age grew at similar rates and manifested similar morphology. However, they responded differently to the applied stressors, even before the production of fetal sex hormones. Furthermore, microarray and qPCR analyses of the whole 10.5 PC embryos also revealed differences in gene expression between male and female tissues. Likewise, the exposure of cells isolated from fetuses and adolescent mice to the stressors and/or sex hormones yielded expression patterns that reflected chromosomal sex, with ethanol feminizing male cells and masculinizing female cells. We conclude that cells differ innately according to sex irrespective of their history of exposure to sex hormones. These differences may have consequences in the course of sexually dimorphic diseases and their therapy. PMID- 19190083 TI - Characterization of exosome-like vesicles released from human tracheobronchial ciliated epithelium: a possible role in innate defense. AB - Airway mucus forms the structural basis of the local innate immune defense mechanism. It is an integrated, active, viscoelastic gel matrix evolved to protect the exposed lung from physical, chemical, and pathological erosion. Exosomes are biologically active vesicles secreted by different cell types including epithelial, hematopoietic, and some tumor cells. They are also present in some biological fluids such as serum, urine, breast milk, and bronchoalveolar lavage fluid. In this study, we demonstrate for the first time that exosome-like vesicles with antiviral properties are present in human tracheobronchial epithelial (HTBE) cell culture secretions. These vesicles have been isolated by differential centrifugation and are characterized further by mass spectrometry, flow cytometry, immunoblotting, electron microscopy, and light-scattering methods. HTBE vesicles exhibited characteristic exosomal size (30-100 nm) and morphology (cup-shaped) with a buoyant density in sucrose of 1.12-1.18 g/ml. Biochemical characterization further revealed typical surface, cytoskeletal, and cytoplasmic proteins characteristic of exosomes, including the multivesicular and late endosomal membrane markers Tsg101 and CD63. The presence of RNA was also observed. The epithelial mucins MUC1, MUC4, and MUC16 also contributed to the vesicles' structure. Notably, alpha-2,6-linked sialic acid was associated with these mucin molecules and subsequent functional analysis showed that these vesicles have a neutralizing effect on human influenza virus, which is known to bind sialic acid. Taken together, these findings suggest that airway epithelial cells release exosome-like vesicles and that these structures may be involved in diverse physiological processes in airway biology, including innate mucosal defense. PMID- 19190084 TI - GM-CSF and IL-4 synergistically trigger dendritic cells to acquire retinoic acid producing capacity. AB - Retinoic acid (RA) produced by intestinal dendritic cells (DCs) imprints gut homing specificity on lymphocytes and enhances Foxp3(+) regulatory T-cell differentiation. The expression of aldehyde dehydrogenase (ALDH) 1A in these DCs is essential for the RA production. However, it remains unclear how the steady state ALDH1A expression is induced under specific pathogen-free (SPF) conditions. Here, we found that bone marrow-derived dendritic cells (BM-DCs) generated with granulocyte-macrophage colony-stimulating factor (GM-CSF) expressed Aldh1a2, an isoform of Aldh1a, but that fms-related tyrosine kinase 3 ligand-generated BM-DCs did not. DCs from mesenteric lymph nodes (MLN) and Peyer's patches (PP) of normal SPF mice expressed ALDH1A2, but not the other known RA-producing enzymes. Employing a flow cytometric method, we detected ALDH activities in 10-30% of PP DCs and MLN-DCs. They were CD11c(high)CD4(-/low)CD8alpha(intermediate)CD11b( /low) F4/80(low/intermediate)CD45RB(low)CD86(high)MHC class II(high)B220( )CD103(+). Equivalent levels of aldehyde dehydrogenase activity (ALDHact) and ALDH1A2 expression were induced synergistically by GM-CSF and IL-4 in splenic DCs in vitro. In BM-DCs, however, additional signals via Toll-like receptors or RA receptors were required for inducing the equivalent levels. The generated ALDH1A2(+) DCs triggered T cells to express gut-homing receptors or Foxp3. GM-CSF receptor-deficient or vitamin A-deficient mice exhibited marked reductions in the ALDHact in intestinal DCs and the T cell number in the intestinal lamina propria, whereas IL-4 receptor-mediated signals were dispensable. GM-CSF(+)CD11c( )F4/80(+) cells existed constitutively in the intestinal tissues. The results suggest that GM-CSF and RA itself are pivotal among multiple microenvironment factors that enable intestinal DCs to produce RA. PMID- 19190085 TI - Fibulin-6 expression and anoikis in human salivary gland epithelial cells: implications in Sjogren's syndrome. AB - Important changes in acinar and ductal morphology and function, together with pronounced extracellular matrix (ECM) remodelling, are detectable in the labial salivary glands of Sjogren's syndrome (SS) patients. The objective of this work was to determine the effect of treatment with the anti-Ro/SSA auto-antibodies, characterizing SS, on the expression of fibulin-6, a member of the fibulins family of the ECM, in primary human salivary gland epithelial cell (SGEC) cultures established from biopsies of labial minor salivary glands obtained from healthy donors. The induction of cell detachment and anoikis in SGECs treated with anti-Ro/SSA auto-antibodies were also investigated. Changes in fibulin-6 mRNA expression were measured by semi-quantitative reverse transcriptase-PCR and real-time PCR. Fibulin-6 expression in cells treated with anti-Ro/SSA auto antibodies was evaluated by flow cytometric analysis and confocal laser scanning microscopy. SGECs undergoing death by anoikis were identified and quantified using Calcein blue/YOPRO-1 dyes. Herein, we present the first evidence of fibulin 6 expression in SGEC that results distributed in the cytoplasm surrounding the inner side of the plasma membrane. Fibulin-6 was down-regulated in SGECs treated with anti-Ro/SSA auto-antibodies in which a marked cell detachment and a reduction of cell viability were observed. Notably, a reduction of fibulin-6 expression in SGECs treated with anti-Ro/SSA auto-antibodies corresponds to an increase of anoikis cell death. Our observations demonstrate a dysregulation of fibulin-6 in the pathological processes observed in SS and provide new evidence that disorganization of the ECM environment could damage the architecture and function of the salivary glands. PMID- 19190086 TI - Variations of radon and thoron concentrations in different types of dwellings in Mysore city, India. AB - (222)Rn and (220)Rn, the immediate decay products of radium isotopes, are causative agents of lung cancer. (222)Rn and (220)Rn concentrations in houses with different floorings, roofs and walls and in different rooms of houses have been measured in Mysore city, Karnataka state, India, using solid-state nuclear track detectors. The radon and thoron concentrations in dwellings with granite floorings are found to be higher compared with other types. A correlation between the indoor radon concentration and dose in air from terrestrial gamma radiation is observed. PMID- 19190087 TI - Feasibility of iterative learning control mediated by functional electrical stimulation for reaching after stroke. AB - BACKGROUND: An inability to perform tasks involving reaching is a common problem following stroke. Evidence supports the use of robotic therapy and functional electrical stimulation (FES) to reduce upper limb impairments, but current systems may not encourage maximal voluntary contribution from the participant because assistance is not responsive to performance. OBJECTIVE: This study aimed to investigate whether iterative learning control (ILC) mediated by FES is a feasible intervention in upper limb stroke rehabilitation. METHODS: Five hemiparetic participants with reduced upper limb function who were at least 6 months poststroke were recruited from the community. No participants withdrew. INTERVENTION: Participants undertook supported tracking tasks using 27 different trajectories augmented by responsive FES to their triceps brachii muscle, with their hand movement constrained in a 2-dimensional plane by a robot. Eighteen 1 hour treatment sessions were used with 2 participants receiving an additional 7 treatment sessions. OUTCOME MEASURES: The primary functional outcome measure was the Action Research Arm Test (ARAT). Impairment measures included the upper limb Fugl-Meyer Assessment (FMA), tests of motor control (tracking accuracy), and isometric force. RESULTS: Compliance was excellent and there were no adverse events. Statistically significant improvements were measured (P 7000 were observed in mock selections of model libraries containing functional riboswitches in liquid culture. The technique was applied to optimize various parameters affecting the selection outcome, and to isolate novel thiamine pyrophosphate riboswitches from a complex library. Artificial riboswitches with excellent characteristics were isolated that exhibit up to 58-fold activation as measured by fluorescent reporter gene assay. PMID- 19190096 TI - Crystal structure of KorA bound to operator DNA: insight into repressor cooperation in RP4 gene regulation. AB - KorA is a global repressor in RP4 which regulates cooperatively the expression of plasmid genes whose products are involved in replication, conjugative transfer and stable inheritance. The structure of KorA bound to an 18-bp DNA duplex that contains the symmetric operator sequence and incorporates 5-bromo-deoxyuridine nucleosides has been determined by multiple-wavelength anomalous diffraction phasing at 1.96-A resolution. KorA is present as a symmetric dimer and contacts DNA via a helix-turn-helix motif. Each half-site of the symmetric operator DNA binds one copy of the protein in the major groove. As confirmed by mutagenesis, recognition specificity is based on two KorA side chains forming hydrogen bonds to four bases within each operator half-site. KorA has a unique dimerization module shared by the RP4 proteins TrbA and KlcB. We propose that these proteins cooperate with the global RP4 repressor KorB in a similar manner via this dimerization module and thus regulate RP4 inheritance. PMID- 19190097 TI - Characterization of pullulanase (PUL)-deficient mutants of rice (Oryza sativa L.) and the function of PUL on starch biosynthesis in the developing rice endosperm. AB - Rice (Oryza sativa) allelic sugary1 (sug1) mutants defective in isoamylase 1 (ISA1) accumulate varying levels of starch and phytoglycogen in their endosperm, and the activity of a pullulanase-type of a debranching enzyme (PUL) was found to correlate closely with the severity of the sug1 phenotype. Thus, three PUL deficient mutants were generated to investigate the function of PUL in starch biosynthesis. The reduction of PUL activity had no pleiotropic effects on the other enzymes involved in starch biosynthesis. The short chains (DP < or = 13) of amylopectin in PUL mutants were increased compared with that of the wild type, but the extent of the changes was much smaller than that of sug1 mutants. The alpha-glucan composition [amylose, amylopectin, water-soluble polysaccharide (WSP)] and the structure of the starch components (amylose and amylopectin) of the PUL mutants were essentially the same, although the average chain length of the B(2-3) chains of amylopectin in the PUL mutant was approximately 3 residues longer than that of the wild type. The double mutants between the PUL-null and mild sug1 mutants still retained starch in the outer layer of endosperm tissue, while the amounts of WSP and short chains (DP < or = 7) of amylopectin were higher than those of the sug1 mutant; this indicates that the PUL function partially overlaps with that of ISA1 and its deficiency has a much smaller effect on the synthesis of amylopectin than ISA1 deficiency and the variation of the sug1 phenotype is not significantly dependent on the PUL activities. PMID- 19190099 TI - Preferences of cancer patients regarding communication of bad news: a systematic literature review. AB - Most physicians regard the communication of bad news to be a difficult issue in clinical oncology practice. The optimal manner of communicating bad news to patients so that physicians can create maximal understanding in patients and facilitate their psychological adjustment is unknown. A systematic review of the literature was conducted to clarify available knowledge on patient preferences regarding the communication of bad news and associated factors. A comprehensive computer search of databases (MEDLINE and PsychINFO) and a manual search identified 24 studies. The above issue has been discussed mainly in Western countries. Most studies used different measures to obtain information on patient preferences and have provided mostly descriptive evidence. The findings in this review suggest that patient preferences with regard to the communication of bad news by physicians consist of four components: setting, manner of communicating bad news, what and how much information is provided and emotional support, and that patients' preferences are associated with demographic factors. Younger patients, female patients and more highly educated patients consistently expressed a desire to receive as much detailed information as possible and to receive emotional support. Asian patients were shown to prefer that relatives be present when receiving bad news more than Westerners do and to prefer to discuss their life expectancy less than Westerners. Physicians need to recognize these preferences to help patients understand. PMID- 19190098 TI - A naturally occurring splicing site mutation in the Brassica rapa FLC1 gene is associated with variation in flowering time. AB - FLOWERING LOCUS C (FLC), encoding a MADS-domain transcription factor in Arabidopsis, is a repressor of flowering involved in the vernalization pathway. This provides a good reference for Brassica species. Genomes of Brassica species contain several FLC homologues and several of these colocalize with flowering time QTL. Here the analysis of sequence variation of BrFLC1 in Brassica rapa and its association with the flowering-time phenotype is reported. The analysis revealed that a G-->A polymorphism at the 5' splice site in intron 6 of BrFLC1 is associated with flowering phenotype. Three BrFLC1 alleles with alternative splicing patterns, including two with different parts of intron 6 retained and one with the entire exon 6 excluded from the transcript, were identified in addition to alleles with normal splicing. It was inferred that aberrant splicing of the pre-mRNA leads to loss-of-function of BrFLC1. A CAPS marker was developed for this locus to distinguish Pi6+1(G) and Pi6+1(A). The polymorphism detected with this marker was significantly associated with flowering time in a collection of 121 B. rapa accessions and in a segregating Chinese cabbage doubled-haploid population. These findings suggest that a naturally occurring splicing mutation in the BrFLC1 gene contributes greatly to flowering-time variation in B. rapa. PMID- 19190100 TI - Identification and molecular characterization of triple- and quadruple-resistant Helicobacter pylori clinical isolates in Germany. AB - OBJECTIVES: The aim of this study was to estimate the frequency of triple- and quadruple-resistant Helicobacter pylori isolated in Germany, to characterize those isolates molecular genetically and to identify risk factors for the development of multiresistance. METHODS: Antimicrobial susceptibility to metronidazole, clarithromycin, amoxicillin, tetracycline, ciprofloxacin/levofloxacin and rifampicin in 1118 clinical isolates obtained between July 2006 and December 2007 was tested by the Etest method. For patients harbouring triple- or quadruple-resistant strains (n = 169), data on prior eradication therapies and underlying diseases were collected and evaluated. A select number of quadruple- and triple-resistant strains were examined for resistance-mediating mutations in their 23S rRNA, 16S rRNA, gyrA and rpoB genes, respectively. RESULTS: From 1118 clinical isolates, 13.4% (n = 150) showed phenotypic resistance to metronidazole, clarithromycin and quinolones and 0.9% (n = 10) to metronidazole, clarithromycin and rifampicin; one isolate exhibited resistance to clarithromycin, quinolones and rifampicin. In eight isolates (0.7%), we detected phenotypic quadruple resistance to metronidazole, clarithromycin, quinolones and rifampicin or tetracycline. Triple- and quadruple resistant strains harboured resistance-associated mutations in their 23S rRNA, 16S rRNA, gyrA or rpoB genes and were nearly exclusively isolated from patients who had already been unsuccessfully treated on multiple occasions. CONCLUSIONS: We show that more than 15% of H. pylori strains isolated from routine samples in the German National Reference Centre are resistant to three or more antimicrobials and identified prior unsuccessful eradication therapies as a key factor for the development of multiresistance. Our data emphasize the need for further comprehensive surveillance studies monitoring the role of treatment regimens in antimicrobial resistance in H. pylori. PMID- 19190101 TI - Comment on: Urinary tract infection in hospitalized elderly patients in the United Kingdom: the importance of making an accurate diagnosis in the post broad spectrum antibiotic era. PMID- 19190102 TI - Localization of insulinomas to regions of the pancreas by intraarterial calcium stimulation: the NIH experience. AB - CONTEXT: Selective intraarterial calcium injection of the major pancreatic arteries with hepatic venous sampling [calcium arterial stimulation (CaStim)] has been used as a localizing tool for insulinomas at the National Institutes of Health (NIH) since 1989. The accuracy of this technique for localizing insulinomas was reported for all cases until 1996. OBJECTIVES: The aim of the study was to assess the accuracy and track record of the CaStim over time and in the context of evolving technology and to review issues related to result interpretation and procedure complications. CaStim was the only invasive preoperative localization modality used at our center. Endoscopic ultrasound (US) was not studied. DESIGN AND SETTING: We conducted a retrospective case review at a referral center. PATIENTS: Twenty-nine women and 16 men (mean age, 47 yr; range, 13-78) were diagnosed with an insulinoma from 1996-2008. INTERVENTION: A supervised fast was conducted to confirm the diagnosis of insulinoma. US, computed tomography (CT), magnetic resonance imaging (MRI), and CaStim were used as preoperative localization studies. Localization predicted by each preoperative test was compared to surgical localization for accuracy. MAIN OUTCOME: We measured the accuracy of US, CT, MRI, and CaStim for localization of insulinomas preoperatively. RESULTS: All 45 patients had surgically proven insulinomas. Thirty-eight of 45 (84%) localized to the correct anatomical region by CaStim. In five of 45 (11%) patients, the CaStim was falsely negative. Two of 45 (4%) had false-positive localizations. CONCLUSION: The CaStim has remained vastly superior to abdominal US, CT, or MRI over time as a preoperative localizing tool for insulinomas. The utility of the CaStim for this purpose and in this setting is thus validated. PMID- 19190103 TI - Cardiovascular and cerebrovascular comorbidities of hypokalemic and normokalemic primary aldosteronism: results of the German Conn's Registry. AB - CONTEXT: Primary aldosteronism (PA) is associated with vascular end-organ damage. OBJECTIVE: Our objective was to evaluate differences regarding comorbidities between the hypokalemic and normokalemic form of PA. DESIGN AND SETTING: This was a retrospective cross-sectional study collected from six German centers (German Conn's registry) between 1990 and 2007. PATIENTS: Of 640 registered patients with PA, 553 patients were analyzed. MAIN OUTCOME MEASURES: Comorbidities depending on hypokalemia or normokalemia were examined. RESULTS: Of the 553 patients (61 +/- 13 yr, range 13-96), 56.1% had hypokalemic PA. The systolic (164 +/- 29 vs. 155 +/- 27 mm Hg; P < 0.01) and diastolic (96 +/- 18 vs. 93 +/- 15 mm Hg; P < 0.05) blood pressures were significantly higher in hypokalemic patients than in those with the normokalemic variant. The prevalence of cardiovascular events (angina pectoris, myocardial infarction, chronic cardiac insufficiency, coronary angioplasty) was 16.3%. Atrial fibrillation occurred in 7.1% and other atrial or ventricular arrhythmia in 5.2% of the patients. Angina pectoris and chronic cardiac insufficiency were significantly more prevalent in hypokalemic PA (9.0 vs. 2.1%, P < 0.001; 5.5 vs. 2.1%, P < 0.01). Overall, cerebrovascular comorbidities were not different between hypokalemic and normokalemic patients, however, stroke tended to be more prevalent in normokalemic patients. CONCLUSIONS: Our data indicate a high prevalence of comorbidities in patients with PA. The hypokalemic variant is defined by a higher morbidity than the normokalemic variant regarding some cardiovascular but not cerebrovascular events. Thus, PA should be sought not only in hypokalemic but also in normokalemic hypertensives because high-excess morbidity occurs in both subgroups. PMID- 19190104 TI - Twenty-four-hour simultaneous subcutaneous Basal-bolus administration of insulin and amylin in adolescents with type 1 diabetes decreases postprandial hyperglycemia. AB - CONTEXT: The purpose of this study was to examine the effect of continuous sc replacement of amylin and insulin for a 24-h period on glucose homeostasis in adolescents with type 1diabetes. METHODS: Thirteen adolescents with type 1 diabetes on insulin pump therapy participated in a randomized, controlled, crossover design study comparing continuous sc insulin monotherapy (part A) vs. continuous sc insulin and pramlintide infusion (part B). In part A, basal and bolus insulin infusion was per prescribed home regimen. In part B, the basal insulin infusion was the same as part A, but prandial insulin boluses were reduced by 20%. Basal and prandial bolus pramlintide were administered simultaneously via another pump. All boluses were given as a dual wave. RESULTS: The study regimen resulted in a 26% reduction in postprandial hyperglycemia as compared to insulin monotherapy (area under the curve, 600 min, 2610 +/- 539 vs. 692 +/- 861 mg/liter . min) (P < 0.008). Glucagon concentrations were suppressed postprandially (P < 0.003) but not in the postabsorptive state, whereas plasma insulin concentrations were unchanged. CONCLUSIONS: Simultaneous continuous sc pramlintide and insulin infusion has the potential of improving glucose concentrations by way of physiological replacement. PMID- 19190105 TI - Association of high iodine intake with the T1799A BRAF mutation in papillary thyroid cancer. AB - CONTEXT: Epidemiological studies have indicated that high iodine intake might be a risk factor for papillary thyroid cancer (PTC), which commonly harbors the oncogenic T1799A BRAF mutation. OBJECTIVE: The objective of the study was to investigate the relationship between BRAF mutation in PTC and iodine intake in patients. SUBJECTS AND METHODS: We analyzed and compared the prevalences of the T1799A BRAF mutation in classical PTC of 1032 patients from five regions in China that uniquely harbor different iodine contents in natural drinking water, ranging from normal (10-21 microg/liter) to high (104-287 microg/liter). The BRAF mutation was identified by direct DNA sequencing. RESULTS: The prevalence of BRAF mutation was significantly higher in any of the regions with high iodine content than any of the regions with normal iodine content. Overall, BRAF mutation was found in 387 of 559 PTC with high iodine content (69%) vs. 252 of 473 PTC with normal iodine content (53%), with an odds ratio of 1.97 (95% confidence interval 1.53-2.55) for the association of BRAF mutation with high iodine content (P < 0.0001). In addition, clinicopathological correlation analysis, the largest one of its type ever, showed that BRAF mutation was significantly associated with extrathyroidal invasion, lymph node metastasis, and advanced tumor stages of PTC. CONCLUSIONS: High iodine intake seems to be a significant risk factor for the occurrence of BRAF mutation in thyroid gland and may therefore be a risk factor for the development of PTC. This large study also confirmed the association of BRAF mutation with poorer clinicopathological outcomes of PTC. PMID- 19190107 TI - Bromocriptine reduces augmented thyrotropin secretion in obese premenopausal women. AB - CONTEXT: Diurnal TSH secretion is enhanced in obese premenopausal women. Dopamine inhibits TSH secretion through activation of dopamine D(2) receptors (D(2)R). Dopamine D(2)R availability in the brain is reduced in obese humans in proportion to body adiposity. We hypothesized that deficient dopamine D(2)R signaling is involved in the enhanced TSH secretion associated with obesity. OBJECTIVE: The effect of short-term bromocriptine treatment on spontaneous TSH secretion in obese women was studied while body weight and caloric intake remained constant. DESIGN AND SETTING: We conducted a prospective, fixed-order, crossover study in a Clinical Research Center. PARTICIPANTS: Seventeen obese women (body mass index, 33.2 +/- 0.6 kg/m(2)) were studied twice in the early follicular phase of their menstrual cycle. INTERVENTION: Subjects were treated for 8 d with placebo and bromocriptine. MAIN OUTCOME MEASURE(S): Blood was collected for 24 h at 10-min intervals, and TSH and leptin were analyzed with deconvolution and correlation techniques, approximate entropy, and cosine regression. RESULTS: Bromocriptine reduced 24-h TSH secretion (placebo, 29.8 +/- 4.6 mU/liter . 24 h, vs. bromocriptine, 22.4 +/- 3.7 mU/liter . 24 h; P = 0.001), whereas free T(4) and total T(3) concentrations did not change. Bromocriptine administration reduced the mesor and amplitude of the 24-h rhythm without resetting the phase. The regularity of the subordinate TSH pattern and synchrony between leptin and TSH were unaffected by bromocriptine. CONCLUSION: Activation of dopamine D(2)R by bromocriptine reverses enhanced diurnal TSH secretion in obese women. Thus, reduced dopaminergic neuronal signaling might be involved in the perturbation of the thyrotrope hormonal axis in obese premenopausal women. PMID- 19190106 TI - Knockdown of IG20 gene expression renders thyroid cancer cells susceptible to apoptosis. AB - AIM: The aim of the study was to investigate the expression and function of the IG20 gene in thyroid cancer cell survival, proliferation, and apoptosis. METHODS: We determined the expression levels of the major isoforms of IG20 by quantitative RT-PCR in normal and thyroid tumor tissues/cell lines. We evaluated the functional consequence of IG20 knockdown in WRO (follicular carcinoma) and FRO (anaplastic carcinoma) thyroid cancer cell lines by measuring spontaneous, TNFalpha-related apoptosis-inducing ligand (TRAIL), and TNFalpha-induced apoptosis. RESULTS: The IG20 gene expression levels were higher in benign and malignant thyroid tumors and in WRO and FRO cells relative to normal tissues. Predominantly, MADD and DENN-SV isoforms of IG20 gene were expressed. IG20 knockdown resulted in increased spontaneous, TRAIL-, and TNFalpha-induced apoptosis in WRO, but not FRO, cells. FRO cell resistance to apoptosis is likely due to caspase-8 deficiency. CONCLUSION: IG20 knockdown renders WRO cells more susceptible to spontaneous, TRAIL-, and TNFalpha-induced apoptosis and thus demonstrates the prosurvival function of the IG20 gene in thyroid cancer. These observations, combined with overexpression of IG20 noted in thyroid tumor tissues, may suggest a potential role in thyroid cancer survival and growth and indicate that IG20 may be targeted either alone or in conjunction with TRAIL or TNFalpha treatment in certain thyroid cancers. PMID- 19190108 TI - 18F-Fluorodeoxyglucose positron emission tomography for the diagnosis of adrenocortical tumors: a prospective study in 77 operated patients. AB - CONTEXT: Most adrenal incidentalomas are nonfunctioning adrenocortical adenomas (ACAs). Adrenocortical carcinomas (ACCs) are rare but should be recognized at an early stage. OBJECTIVE: The objective of the study was to evaluate the usefulness of (18)F-fluorodeoxyglucose positron emission tomography ((18)F-FDG PET) to predict malignancy in patients without a previous history of cancer. DESIGN: This was a prospective, multicenter study from 2001 to 2006. SETTING: The study was conducted at a network of seven university hospitals in Paris. PATIENTS: Seventy seven patients were included. All underwent surgery because of hypersecretory and/or growing benign lesions (n = 18), obvious ACCs (n = 21), or radiologically indeterminate lesions (n = 38). MAIN OUTCOME MEASURE: The degree of (18)F-FDG PET uptake [maximum standardized uptake value (maxSUV)] was related to the pathological findings serving as a reference, and its diagnostic value was compared with that of computerized tomography (CT) scan. RESULTS: Pathology eventually diagnosed 43 ACAs, 22 ACCs, and 12 nonadrenocortical lesions. Using a cutoff value above 1.45 for adrenal to liver maxSUV ratio, the sensitivity and specificity to distinguish ACAs from ACCs were, respectively, 1.00 (95% confidence interval 0.85-1.00) and 0.88 (95% confidence interval 0.75-0.96). Among the 38 indeterminate lesions at CT scan, we could analyze a subgroup of 16 adrenocortical tumors with high unenhanced density (>10 HU) and an inappropriate washout: (18)F-FDG PET correctly predicted the benignity in 13 of 15 ACAs. CONCLUSIONS: In a multidisciplinary team approach, (18)F-FDG PET helps to manage suspicious CT scan lesions. An adrenal to liver maxSUV ratio less than 1.45 is highly predictive of a benign lesion. PMID- 19190109 TI - Type I gonadotropin-releasing hormone receptor mediates the antiproliferative effects of GnRH-II on prostate cancer cells. AB - BACKGROUND: GnRH-II has been shown to exert a strong antiproliferative action on tumors of the female reproductive system. The data so far reported on the effects of GnRH-II on prostate cancer growth are controversial. Moreover, it is still unclear through which receptor [type I or type II GnRH-receptor (GnRH-R)] GnRH-II might modulate cancer cell proliferation. OBJECTIVE: The objective of this work was to investigate whether GnRH-II might affect the proliferation of prostate cancer cells and to identify the GnRH-R through which the peptide might exert its activity. DESIGN: We investigated the effects of GnRH-II on prostate cancer cell proliferation. We then transfected PC3 cells with a small interfering RNA targeted to type I GnRH-R. After receptor silencing we evaluated the effects of GnRH-II on cell proliferation and on forskolin-induced intracellular cAMP accumulation. Similar experiments were performed by silencing type II GnRH-R. RESULTS: GnRH-II exerted an antiproliferative activity on prostate cancer cells. Transfection of PC3 cells with a type I GnRH-R small interfering RNA resulted in a significant decrease of the expression of this receptor. After type I GnRH-R silencing: 1) the antiproliferative effect of GnRH-II was completely abrogated; and 2) GnRH-II lost its capacity to counteract the forskolin-induced cAMP accumulation. On the contrary, type II GnRH-R silencing did not counteract the antiproliferative effect of GnRH-II. CONCLUSIONS: GnRH-II exerts a specific and significant antiproliferative action on prostate cancer cells. This antitumor effect is mediated by the activation of type I (but not of type II) GnRH-R and by its coupled cAMP intracellular signaling pathway. PMID- 19190111 TI - Euthyroid hyperthyrotropinemia in children born after in vitro fertilization. AB - CONTEXT: Assisted reproduction techniques are now commonly used. Although classic in vitro fertilization (IVF) started almost 30 yr ago, few long-term systematic prospective studies of children conceived with assisted reproduction have been performed. OBJECTIVE: Our objective was to investigate thyroid function in children conceived after IVF vs. naturally conceived controls. POPULATIONS AND METHODS: A total of 106 children conceived after classic IVF and 68 naturally conceived controls, aged 4-14 yr, were studied. All children were thoroughly examined, and serum T(3), T(4), TSH, anti-thyroid peroxidase, and anti thyroglobulin antibodies were measured. A second TSH determination and a thyroid ultrasound were performed for TSH higher than 5 microIU/ml, and children were considered to have persistent hyperthyrotropinemia, if the TSH elevation was confirmed. RESULTS: Seven IVF children but none of the controls had persistent elevations of circulating TSH, suggesting euthyroid hyperthyrotropinemia or subclinical primary hypothyroidism (P = 0.044). TSH was significantly higher in the IVF group than in controls (P = 0.046), whereas no significant differences in the concentrations of T(3) or T(4) were observed. None of the children had detectable circulating antithyroid antibodies in either group. CONCLUSIONS: A significant elevation of serum TSH compatible with a mild TSH resistance of the thyroid were found in IVF children compared with controls. This was not due to the presence of antithyroid autoantibodies. We suggest that this might represent a slight epigenetic developmental abnormality related to the preimplantation manipulation of the embryo. Further studies are needed to confirm these findings and to better determine their etiopathogenesis and clinical significance. PMID- 19190110 TI - Blunted sleep-related luteinizing hormone rise in healthy premenarcheal pubertal girls with elevated body mass index. AB - OBJECTIVE: Our objective was to determine whether excessive adiposity is associated with alteration of the normal hormonal changes of early pubertal girls. DESIGN AND PARTICIPANTS: Healthy 6.4- to 9.5-yr-old, prepubertal (PRE, n = 20) and 9.4- to 13.0-yr-old pubertal premenarcheal volunteers (PUB, n = 20) were divided into excessive-weight (EW) or normal-weight (NW) groups according to the 85th percentile body mass index. INTERVENTIONS AND SETTING: Overnight blood sampling; GnRH agonist (GnRHag), low-dose ACTH, oral glucose tolerance tests, and pelvic ultrasonograms were performed in our Clinical Research Center. RESULTS: EW girls were similar in age and baseline and ACTH- and GnRHag-stimulated androgen levels to stage-matched NW girls. However, the sleep-related LH rise was blunted in EW-PUB girls compared with NW-PUB girls. The sleep-related rise of mean LH in EW-PUB [0.68 +/- 0.35 (sem) U/liter] was insignificant, less than that of NW-PUB (2.1 +/- 0.45, P < 0.05) and not significantly different from that of PRE girls (0.08+/-0.03). EW-PUB had slower LH pulse frequency and a lower rise in LH pulse amplitude during sleep than NW-PUB girls (both P < 0.05). Overnight FSH patterns paralleled LH patterns, whereas estradiol levels were similar in stage-matched NW and EW groups, differing between stages as expected. Early morning and peak LH, FSH, and estradiol responses to GnRHag were similar in EW-PUB and NW-PUB and significantly greater than those of PRE girls. CONCLUSIONS: Healthy EW-PUB girls have significantly blunted sleep-related LH production. These data suggest that excess adiposity, in the absence of sex steroid excess, may subtly suppress hypothalamic-pituitary-gonadal function in premenarcheal pubertal girls. PMID- 19190112 TI - Maternal levels of corticotropin-releasing hormone during pregnancy in relation to adiponectin and leptin in early childhood. AB - BACKGROUND: Fetal glucocorticoid exposure is associated with later development of features of the metabolic syndrome such as central obesity and insulin resistance. Fat tissue, especially visceral fat, produces adiponectin, which is inversely associated with insulin resistance in older children and adults. Adipocytes also produce leptin, directly related to measures of adiposity. It is unknown how the secretion of these hormones in early childhood is related to pregnancy levels of CRH, a proxy of fetal glucocorticoid exposure. AIM: Our aim was to study the relationship of maternal midpregnancy CRH levels with offspring levels of adiponectin and leptin in early childhood. METHODS: The study population consisted of 349 mother-children pairs from Project Viva, a prospective prebirth cohort study from eastern Massachusetts. We created a general linear model with log CRH levels in midpregnancy maternal blood as the predictor and adiponectin and leptin measured in the 3-yr-old offspring as outcomes, adjusting for covariates. RESULTS: The means (sd) of log CRH, adiponectin, and leptin were 4.97 (0.65) log pg/ml, 22.4 (5.8) microg/ml, and 1.9 (1.8) ng/ml. For each unit increment in log CRH, mean value of offspring adiponectin was 1.10 microg/ml (95% confidence interval = 0.06-2.14) higher. We found no association with leptin (-0.08 ng/ml; 95% confidence interval = -0.40 0.24). CONCLUSIONS: Higher maternal blood levels of CRH were associated with higher levels of adiponectin but unchanged levels of leptin at age 3 yr. The increased adiponectin levels might represent secretion from organs other than fat or reflect a compensatory mechanism to increase insulin sensitivity. PMID- 19190113 TI - Common variation in the DIO2 gene predicts baseline psychological well-being and response to combination thyroxine plus triiodothyronine therapy in hypothyroid patients. AB - INTRODUCTION: Animal studies suggest that up to 80% of intracellular T(3) in the brain is derived from circulating T(4) by local deiodination. We hypothesized that in patients on T(4) common variants in the deiodinase genes might influence baseline psychological well-being and any improvement on combined T(4)/T(3) without necessarily affecting serum thyroid hormone levels. METHODS: We analyzed common variants in the three deiodinase genes vs. baseline psychological morbidity and response to T(4)/T(3) in 552 subjects on T(4) from the Weston Area T(4) T(3) Study (WATTS). Primary outcome was improvement in psychological well being assessed by the General Health Questionnaire 12 (GHQ-12). RESULTS: The rarer CC genotype of the rs225014 polymorphism in the deiodinase 2 gene (DIO2) was present in 16% of the study population and was associated with worse baseline GHQ scores in patients on T(4) (CC vs. TT genotype: 14.1 vs. 12.8, P = 0.03). In addition, this genotype showed greater improvement on T(4)/T(3) therapy compared with T(4) only by 2.3 GHQ points at 3 months and 1.4 at 12 months (P = 0.03 for repeated measures ANOVA). This polymorphism had no impact on circulating thyroid hormone levels. CONCLUSIONS: Our results require replication but suggest that commonly inherited variation in the DIO2 gene is associated both with impaired baseline psychological well-being on T(4) and enhanced response to combination T(4)/T(3) therapy, but did not affect serum thyroid hormone levels. PMID- 19190114 TI - MAPK-independent impairment of T-cell responses by the multikinase inhibitor sorafenib. AB - Sorafenib, originally developed as CRAF inhibitor but soon recognized as a multikinase inhibitor, is currently widely tested for the treatment of different cancers either alone or in combination therapy. However, the clinical success, particularly in immunogenic tumors such as melanoma, was less than anticipated. Because T-cell activation is tightly regulated by a multitude of kinases, we scrutinized effects of sorafenib on immune responses. To this end, comprehensive in vitro studies revealed that the presence of sorafenib concentrations comparable with observed plasma levels in patients strongly impairs the activation of T cells. Notably, even established tumor-specific immune responses are influenced by sorafenib. Indeed, ELISPOT data of peripheral blood lymphocytes obtained from melanoma patients vaccinated against survivin show markedly diminished survivin-specific immune responses in the presence of sorafenib. Surprisingly, inhibition of T-cell activation was not associated with reduced extracellular signal-regulated kinase phosphorylation. In fact, on T-cell receptor stimulation phospho-extracellular signal-regulated kinase and phospho mitogen-activated protein kinase kinase levels were found to be elevated in the presence of sorafenib, showing the complexity of signal transduction events following T-cell receptor stimulation. In conclusion, our data show that T-cell function is sensitive toward the multikinase inhibitor sorafenib in a mitogen activated protein kinase-independent fashion. This observation has important implications for the use of sorafenib as therapy for immunogenic cancers. PMID- 19190115 TI - Mechanisms involved in synergistic anticancer effects of anti-4-1BB and cyclophosphamide therapy. AB - Chemotherapy can precondition for immunotherapy by creating an environment for homeostatic lymphoproliferation and eliminating some of the suppressive immune networks. We found that combination therapy with anti-4-1BB and cyclophosphamide (CTX) produced synergistic anticancer effects in the poorly immunogenic B16 melanoma model in mice. The antitumor effect of the combination therapy depended mainly on CD8(+) T cells, the 4-1BB-dependent expansion and differentiation of which into IFN-gamma-producing CD11c(+)CD8(+) T cells was enhanced by CTX. Anti-4 1BB induced a rapid repopulation of T and B cells from CTX-mediated lymphopenia. Anti-4-1BB protected naive T cells from CTX and promoted proliferation of memory/effector and memory T cells. The combination treatment produced approximately 60- and 2.2-fold more CTLs per tumor-associated antigen compared with CTX or anti-4-1BB alone, respectively. This indicates that anti-4-1BB promoted a preferential expansion of tumor-specific CD8(+) T cells among the repopulated lymphocytes following CTX-mediated lymphopenia. CTX treatment enhanced 4-1BB expression on CD4 and CD8 T cells, and CTX alone or in combination with anti-4-1BB effectively suppressed peripheral regulatory T cells. Our results indicate that anti-4-1BB and CTX can be practical partners in cancer therapy because CTX creates an environment in which anti-4-1BB actively promotes the differentiation and expansion of tumor-specific CTLs. PMID- 19190116 TI - Trabectedin (ET-743) promotes differentiation in myxoid liposarcoma tumors. AB - Differentiation is a complex set of events that can be blocked by rearrangements of regulatory genes producing fusion proteins with altered properties. In the case of myxoid liposarcoma (MLS) tumors, the causative abnormality is a fusion between the CHOP transcription factor and the FUS or EWS genes. CHOP belongs to and is a negative regulator of the large CAAT/enhancer binding protein family whose alpha, beta, and delta members are master genes of adipogenesis. Recent clinical data indicate a peculiar sensitivity of these tumors to the natural marine compound trabectedin. One hypothesis is that the activity of trabectedin is related to the inactivation of the FUS-CHOP oncogene. We find that trabectedin causes detachment of the FUS-CHOP chimera from targeted promoters. Reverse transcription-PCR and chromatin immunoprecipitation analysis in a MLS line and surgical specimens of MLS patients in vivo show activation of the CAAT/enhancer binding protein-mediated transcriptional program that leads to morphologic changes of terminal adipogenesis. The activity is observed in cells with type 1 but not type 8 fusions. Hence, the drug induces maturation of MLS lipoblasts in vivo by targeting the FUS-CHOP-mediated transcriptional block. These data provide a rationale for the specific activity of trabectedin and open the perspective of combinatorial treatments with drugs acting on lipogenic pathways. PMID- 19190117 TI - Dihydrofolate reductase amplification and sensitization to methotrexate of methotrexate-resistant colon cancer cells. AB - Gene amplification is one of the most frequent manifestations of genomic instability in human tumors and plays an important role in tumor progression and acquisition of drug resistance. To better understand the factors involved in acquired resistance to cytotoxic drugs via gene amplification, we have analyzed the structure and dynamics of dihydrofolate reductase (DHFR) gene amplification in HT29 cells treated with methotrexate (MTX). Analysis of the DHFR gene amplification process shows that the amplicon exhibits a complex structure that is consistently reproduced in independent treatments. The cytogenetic manifestation of the amplification in advanced stages of the treatment may be in the form of double minutes or as a homogeneously stained region. To get insights into the mechanisms of resistance, we have also investigated the sensitization to MTX of MTX-resistant cells after drug withdrawal and reexposure to MTX. Passive loss of the DHFR amplicon by withdrawal of the drug results in MTX-sensitive cells exhibiting a substantial reduction of their capacity or even an incapacity to generate resistance when submitted to a second cycle of MTX treatment. On a second round of drug administration, the resistant cells generate a different amplicon structure, suggesting that the formation of the amplicon as in the first cycle of treatment is not feasible. These results indicate that DHFR gene amplification is a "wear and tear" process in HT29 cells and that MTX-resistant cells may become responsive to a second round of treatment if left untreated during a sufficient period of time. PMID- 19190118 TI - CNOB/ChrR6, a new prodrug enzyme cancer chemotherapy. AB - We report the discovery of a new prodrug, 6-chloro-9-nitro-5-oxo-5H benzo(a)phenoxazine (CNOB). This prodrug is efficiently activated by ChrR6, the highly active prodrug activating bacterial enzyme we have previously developed. The CNOB/ChrR6 therapy was effective in killing several cancer cell lines in vitro. It also efficiently treated tumors in mice with up to 40% complete remission. 9-Amino-6-chloro-5H-benzo(a)phenoxazine-5-one (MCHB) was the only product of CNOB reduction by ChrR6. MCHB binds DNA; at nonlethal concentration, it causes cell accumulation in the S phase, and at lethal dose, it induces cell surface Annexin V and caspase-3 and caspase-9 activities. Further, MCHB colocalizes with mitochondria and disrupts their electrochemical potential. Thus, killing by CNOB involves MCHB, which likely induces apoptosis through the mitochondrial pathway. An attractive feature of the CNOB/ChrR6 regimen is that its toxic product, MCHB, is fluorescent. This feature proved helpful in in vitro studies because simple fluorescence measurements provided information on the kinetics of CNOB activation within the cells, MCHB killing mechanism, its generally efficient bystander effect in cells and cell spheroids, and its biodistribution. The emission wavelength of MCHB also permitted its visualization in live animals, allowing noninvasive qualitative imaging of MCHB in mice and the tumor microenvironment. This feature may simplify exploration of barriers to the penetration of MCHB in tumors and their amelioration. PMID- 19190119 TI - Dasatinib-induced autophagy is enhanced in combination with temozolomide in glioma. AB - Glioblastoma is defined by its aggressive invasion, microvascular proliferation, and central necrosis. BMS-354825 (dasatinib) is an ATP-competitive small-molecule inhibitor effective in treating drug-resistant tumors with mutant BCR-ABL, KIT, and epidermal growth factor receptor by blocking tyrosine phosphorylation sites that are critical in tumorigenesis. In studying the action of dasatinib in human glioblastoma, we found that levels of phospho-SRC, AKT, and ribosomal protein S6 were decreased in cell lines treated with low nanomolar concentrations of dasatinib at baseline and following stimulation with epidermal growth factor. Furthermore, an increased sensitivity to dasatinib was noted in glioma cells with functional PTEN. Reduction of invasive potential was observed in vitro at concentrations well below the IC(50) of dasatinib, which was corroborated by immunofluorescence staining showing disruption of paxillin localization to focal adhesions and decreases in focal adhesion kinase autophosphorylation. Cell cycle analysis revealed minimal G(1) arrest but a significant increase in autophagic cell death in glioma cells treated with dasatinib as assessed by acridine orange staining and a concomitant increase in light chain 3 expression and processing. Combination treatment of glioma cells with dasatinib and temozolomide resulted in a significant increase in cell cycle disruption and autophagic cell death. Dasatinib in combination with temozolomide more effectively increased the therapeutic efficacy of temozolomide than when dasatinib was combined with carboplatin or irinotecan. These results strongly support the clinical use of dasatinib in the treatment of glioblastoma and provide a rationale for combination therapy with dasatinib and temozolomide. PMID- 19190120 TI - Interactions between PTEN and the c-Met pathway in glioblastoma and implications for therapy. AB - The tyrosine kinase receptor c-Met and its ligand hepatocyte growth factor (HGF) are frequently overexpressed and the tumor suppressor PTEN is often mutated in glioblastoma. Because PTEN can interact with c-Met-dependent signaling, we studied the effects of PTEN on c-Met-induced malignancy and associated molecular events and assessed the potential therapeutic value of combining PTEN restoration approaches with HGF/c-Met inhibition. We studied the effects of c-Met activation on cell proliferation, cell cycle progression, cell migration, cell invasion, and associated molecular events in the settings of restored or inhibited PTEN expression in glioblastoma cells. We also assessed the experimental therapeutic effects of combining anti-HGF/c-Met approaches with PTEN restoration or mTOR inhibition. PTEN significantly inhibited HGF-induced proliferation, cell cycle progression, migration, and invasion of glioblastoma cells. PTEN attenuated HGF induced changes of signal transduction proteins Akt, GSK-3, JNK, and mTOR as well as cell cycle regulatory proteins p27, cyclin E, and E2F-1. Combining PTEN restoration to PTEN-null glioblastoma cells with c-Met and HGF inhibition additively inhibited tumor cell proliferation and cell cycle progression. Similarly, combining a monoclonal anti-HGF antibody (L2G7) with the mTOR inhibitor rapamycin had additive inhibitory effects on glioblastoma cell proliferation. Systemic in vivo delivery of L2G7 and PTEN restoration as well as systemic in vivo deliveries of L2G7 and rapamycin additively inhibited intracranial glioma xenograft growth. These preclinical studies show for the first time that PTEN loss amplifies c-Met-induced glioblastoma malignancy and suggest that combining anti-HGF/c-Met approaches with PTEN restoration or mTOR inhibition is worth testing in a clinical setting. PMID- 19190121 TI - Notch1 mediates growth suppression of papillary and follicular thyroid cancer cells by histone deacetylase inhibitors. AB - Notch1 is a multifunctional transmembrane receptor that regulates cellular differentiation, development, proliferation, and survival in a variety of contexts. We have previously shown that Notch1 may function as a tumor suppressor and that histone deacetylase (HDAC) inhibitors can induce Notch1 expression in some endocrine cancers. Here, we showed that although there was minimal Notch1 expression in follicular thyroid cancer FTC236 and papillary thyroid cancer DRO cells, transfection of constitutive Notch1 plasmid into these cells led to growth inhibition, down-regulation of cyclin D1, and up-regulation of p21. Treatment of FTC236 cells with HDAC inhibitors valproic acid (1-4 mmol/L) or suberoyl bishydroxamic acid (10-30 micromol/L) induced functional Notch1 protein expression and suppressed cell growth in a dose-dependent manner. Notch1 siRNA interference blocked the antiproliferative effect of HDAC inhibitors. Western blot analysis revealed the reduction of cyclin D1 and the increase of p21 in HDAC inhibitor-treated cells. These results indicate that HDAC inhibitors activate Notch1 signaling in thyroid cancer cells and lead to the suppression of proliferation by cell cycle arrest. Our findings provide the first documentation of the role of Notch1 signaling as a tumor suppressor in DRO and FTC236 cells, suggesting that Notch1 activation may be a potential therapeutic target for papillary and follicular thyroid cancers. PMID- 19190122 TI - Monoclonal antibody 16D10 to the COOH-terminal domain of the feto-acinar pancreatic protein targets pancreatic neoplastic tissues. AB - We have shown that the 16D10 antigen located on the mucin-like COOH-terminal domain of the feto-acinar pancreatic protein (FAPP) is expressed at the surface of human pancreatic tumor cell lines such as SOJ-6 cell line. Furthermore, an in vivo study indicates that targeting this cell-membrane glycopeptide by the use of the monoclonal antibody (mAb) 16D10 inhibits the growth of SOJ-6 xenografts in nude mice. To validate the potential use of the mAb16D10 in immune therapy, this study examined the expression of 16D10 antigens at the surface of human pancreatic adenocarcinomas versus control tissues. We examined the reactivity of mAb16D10 and mAb8H8 with pancreatic ductal adenocarcinomas (PDAC) compared with controls by using immunohistochemistry and confocal laser scanning microscopy. mAb8H8 does react with control or nontumoral human pancreatic tissues. mAb16D10 has a strong and specific reactivity with PDAC and does not react with other cancers of epithelia or normal tissues tested. Notable, mAb16D10 mostly recognizes membrane of tumoral cells. Furthermore, mAb8H8 and mAb16D10 recognized a protein of 110 to 120 kDa in homogenates of nontumoral and tumoral human pancreatic tissues, respectively. This size correlates with that of FAPP or with that of the normal counterpart of FAPP, the so-called bile salt-dependent lipase. The results suggest that mAb16D10 presents a unique specificity against PDAC; consequently, it could be effective in immune therapy of this cancer. Furthermore, mAb16D10 and mAb8H8 pair might be useful for diagnosis purpose in discriminating tumoral from nontumoral human pancreatic tissues. PMID- 19190124 TI - Inhibiting the teratogenicity of the immunosuppressant leflunomide in mice by supplementation of exogenous uridine. AB - Leflunomide is an immunosuppressant drug displaying teratogenicity in mice, rats, and rabbits. Its immunosuppressive effect occurs via inhibition of dihydroorotate dehydrogenase (DHODH) and tyrosine kinases. In this study, we coadministered Leflunomide and uridine, a precursor substance of pyrimidine nucleotides, to pregnant CD-1 mice, and examined whether or not a decreased level of intracellular pyrimidine nucleotides with inhibition of DHODH is related to the teratogenicity of Leflunomide. Then we examined the alteration of the nucleotide level in fetal tissue by Leflunomide and the effect of coadministered uridine. We administered Leflunomide with or without uridine to pregnant mice on gestation day 10, and used the vehicle of Leflunomide as a control. Leflunomide caused multiple malformations in all fetuses, but coadministration with uridine inhibited most of its teratogenicity. Leflunomide decreased the concentration of pyrimidine nucleotides, not purine nucleotides, whereas uridine coadministered with Leflunomide partially restored the level of pyrimidine nucleotides. These results indicate that the inhibitory effect of DHODH activity is related to the teratogenicity of Leflunomide. PMID- 19190123 TI - Evaluation of endocrine disrupting effects of nitrate after in utero exposure in rats and of nitrate and nitrite in the H295R and T-screen assay. AB - Animal studies have shown that nitrate acts as an endocrine disrupter affecting the androgen production in adult males. This raises a concern for more severe endocrine disrupting effects after exposure during the sensitive period of prenatal male sexual development. As there are no existing studies of effects of nitrate on male sexual development, the aim of the study was to examine how in utero exposure to nitrate would affect male rat fetuses. Pregnant dams were dosed with nitrate in the drinking water from gestational day (GD) 7 to GD21 at the following dose levels 17.5, 50, 150, 450, and 900 mg/l. At GD21, fetuses were examined for anogenital distance, plasma thyroxine levels, testicular and plasma levels of testosterone and progesterone, and testicular testosterone production and histopathology. In addition, endocrine disrupting activity of nitrate and nitrite were studied in two in vitro assays, the H295R assay and T-screen. There were no consistent indications that nitrate induces anti-androgenic effects in male fetuses or that prenatal nitrate exposure affected the thyroid axis. However, a more comprehensive study with long-term exposure before and during pre and postnatal development would be relevant to sufficiently address the concerns based on the indications for endocrine disrupting effects in adult animals. PMID- 19190125 TI - Developmental exposure to chlorpyrifos induces alterations in thyroid and thyroid hormone levels without other toxicity signs in CD-1 mice. AB - Organophosphorus insecticides, as Chlorpyrifos (CPF), are widely used in agriculture and against household pests; these compounds receive an increasing consideration as potential endocrine disrupters. The aim of the present study was to examine the potential short- and long-term effects of CPF on thyroid and adrenal glands in CD1 mice following exposure at dose levels not inducing brain acetyl cholinesterase (AchE) inhibition, during gestational and/or postnatal vulnerable phases. Pregnant dams were treated with 0, 3, 6 mg/kg bw/day of CPF on gestational days 15-18. After delivery, pups were treated subcutaneously on postnatal days (PND) 11-14 with: 0, 1, 3 mg/kg bw/day of CPF. Serum thyroxin (T4), thyroid and adrenals histology and histomorphometry were evaluated in dams and in F1 mice. In dams at 6 mg/kg, decreased T4 levels and increased cell height in thyroid were observed, and adrenal histology showed a slightly increased vacuolization in the X-zone. In the F1, short-term morphological modifications (reduced follicular size at PND 2) and long-term morphological (increased necrotic follicular cells) and biochemical alterations (reduced serum T4 levels) were found at PND 150 with an apparent higher vulnerability of males. For the first time these results indicate that CPF exposure at dose levels not inducing brain AchE inhibition causes thyroid alterations in dams and in F1 CD1 mice. Thyroid may be a sensitive target to CPF developmental exposure possibly leading to long-term effects on thyroid function. Because thyroid plays a pivotal role in mammalian development, these findings can be relevant to humans. PMID- 19190126 TI - Small-molecule drugs mimicking DNA damage: a new strategy for sensitizing tumors to radiotherapy. AB - PURPOSE: Enhanced DNA repair activity is often associated with tumor resistance to radiotherapy. We hypothesized that inhibiting DNA damage repair would sensitize tumors to radiation-induced DNA damage. EXPERIMENTAL DESIGN: A novel strategy for inhibiting DNA repair was tested. We designed small DNA molecules that mimic DNA double-strand breaks (called Dbait) and act by disorganizing damage signaling and DNA repair. We analyzed the effects of Dbait in cultured cells and on xenografted tumors growth and performed preliminary studies of their mechanism(s) of action. RESULTS: The selected Dbait molecules activate H2AX phosphorylation in cell culture and in xenografted tumors. In vitro, this activation correlates with the reduction of Nijmegen breakage syndrome 1 and p53 binding protein 1 repair foci formation after irradiation. Cells are sensitized to irradiation and do not efficiently repair DNA damage. In vivo, Dbait induces regression of radioresistant head and neck squamous cell carcinoma (Hep2) and melanoma (SK28 and LU1205) tumors. The combination of Dbait32Hc treatment and fractionated radiotherapy significantly enhanced the therapeutic effect. Tumor growth control by Dbait molecules depended directly on the dose and was observed with various irradiation protocols. The induction of H2AX phosphorylation in tumors treated with Dbait suggests that it acts in vivo through the induction of "false" DNA damage signaling and repair inhibition. CONCLUSIONS: These data validate the concept of introducing small DNA molecules, which mimic DNA damage, to trigger "false" signaling of DNA damage and impair DNA repair of damaged chromosomes. This new strategy could provide a new method for enhancing radiotherapy efficiency in radioresistant tumors. PMID- 19190127 TI - Phase I and pharmacokinetic study of pegylated liposomal CKD-602 in patients with advanced malignancies. AB - PURPOSE: S-CKD602 is a pegylated liposomal formulation of CKD602, a semisynthetic camptothecin analogue. Pegylated (STEALTH) liposomes can achieve extended drug exposure in plasma and tumor. Based on promising preclinical data, the first phase I study of S-CKD602 was done in patients with refractory solid tumors. EXPERIMENTAL DESIGN: S-CKD602 was administered i.v. every 3 weeks. Modified Fibonacci escalation was used (three to six patients/cohort), and dose levels ranged from 0.1 to 2.5 mg/m2. Serial plasma samples were obtained over 2 weeks and total (lactone+hydroxyl acid) concentrations of encapsulated, released, and sum total (encapsulated+released) CKD602 measured by liquid chromatography-tandem mass spectrometry. RESULTS: Forty-five patients (21 males) were treated. Median age, 62 years (range, 33-79 years) and Eastern Cooperative Oncology Group status, 0 to 1 (43 patients) and 2 (2 patients). Dose-limiting toxicities of grade 3 mucositis occurred in one of six patients at 0.3 mg/m2, grade 3 and 4 bone marrow suppression in two of three patients at 2.5 mg/m2, and grade 3 febrile neutropenia and anemia in one of six patients at 2.1 mg/m2. The maximum tolerated dose was 2.1 mg/m2. Partial responses occurred in two patients with refractory ovarian cancer (1.7 and 2.1 mg/m2). High interpatient variability occurred in the pharmacokinetic disposition of encapsulated and released CKD602. CONCLUSIONS: S CKD602 represents a promising new liposomal camptothecin analogue with manageable toxicity and promising antitumor activity. Phase II studies of S-CKD602 at 2.1 mg/m2 i.v. once every 3 weeks are planned. Prolonged plasma exposure over 1 to 2 weeks is consistent with STEALTH liposomes and provides extended exposure compared with single doses of nonliposomal camptothecins. PMID- 19190128 TI - Local delivery of poly lactic-co-glycolic acid microspheres containing imatinib mesylate inhibits intracranial xenograft glioma growth. AB - PURPOSE: In an effort to develop new therapeutic strategies to treat malignant gliomas, we have designed poly (lactic-co-glycolic) acid (PLGA) microparticles that deliver imatinib mesylate, a small molecule tyrosine kinase inhibitor. The local continuous release of imatinib mesylate at the tumor site overcomes many obstacles associated with systemic delivery. EXPERIMENTAL DESIGN: Polymeric microspheres were prepared from various compositions of PLGA and loaded with imatinib mesylate. Imatinib release profiles, biological activity, and effect on PDGFR-B phosphorylation were confirmed in vitro. The therapeutic efficacy of imatinib microspheres was examined in two s.c. and orthotopic human glioblastoma xenograft models. RESULTS: A single local injection of PLGA microspheres loaded with a low concentration of imatinib mesylate led to 88% and 79% reduction in s.c. human (U87-MG) and murine (GL261) glioma tumors, respectively. PLGA-imatinib mesylate administered intracranially led to a 79% reduction in U87MG tumor volume. Immunohistochemical analysis showed a marked decrease in proliferation indices and tumor vessel density in the s.c. model and induction of apoptosis in an intracranial model. CONCLUSION: This is the first study to show the therapeutic efficacy of the local delivery of imatinib mesylate using a polymeric delivery system. PMID- 19190130 TI - Longitudinal analysis and prognostic effect of cancer-testis antigen expression in multiple myeloma. AB - PURPOSE: Reliable data on the persistence of tumor expression of cancer-testis (CT) antigens over time and consequent analyses of the effect of CT antigen expression on the clinical course of malignancies are crucial for their evaluation as diagnostic markers and immunotherapeutic targets. EXPERIMENTAL DESIGN: Applying conventional reverse transcription-PCR, real-time PCR, and Western blot, we did the first longitudinal study of CT antigen expression in multiple myeloma analyzing 330 bone marrow samples from 129 patients for the expression of four CT antigens (MAGE-C1/CT7, MAGE-C2/CT10, MAGE-A3, and SSX-2). RESULTS: CT antigens were frequently and surprisingly persistently expressed, indicating that down-regulation of these immunogenic targets does not represent a common tumor escape mechanism in myeloma. We observed strong correlations of CT antigen expression levels with the clinical course of myeloma patients as indicated by the number of bone marrow-residing plasma cells and peripheral paraprotein levels, suggesting a role for CT antigens as independent tumor markers. Investigating the prognostic value of CT antigen expression in myeloma patients after allogeneic stem cell transplantation, we found that expression of genes, such as MAGE-C1, represents an important indicator of early relapse and dramatically reduced survival. CONCLUSIONS: Our findings suggest that CT antigens might promote the progression of multiple myeloma and especially MAGE-C1/CT7, which seems to play the role of a "gatekeeper" gene for other CT antigens, might characterize a more malignant phenotype. Importantly, our study also strongly supports the usefulness of CT antigens as diagnostic and prognostic markers as well as therapeutic targets in myeloma. PMID- 19190131 TI - Marked activity of irinotecan and rapamycin combination toward colon cancer cells in vivo and in vitro is mediated through cooperative modulation of the mammalian target of rapamycin/hypoxia-inducible factor-1alpha axis. AB - PURPOSE: Despite recent progress, colon cancer is often resistant to combination chemotherapy, highlighting the need for development of novel therapeutic approaches. An attractive target is hypoxia-inducible factor-1alpha (HIF-1alpha), a key transcription factor with a pivotal role in tumor cell metabolism. One potential class of therapeutic agents targeting HIF-1alpha are mammalian target of rapamycin inhibitors such as rapamycin. A second class are topoisomerase I inhibitors, such as irinotecan, which are able to inhibit the accumulation of HIF 1alpha. We here investigated whether combination of rapamycin and irinotecan was active in human colon cancer models. EXPERIMENTAL DESIGN: Human metastatic tumors were xenografted in nude mice and treated with low doses of irinotecan alone, rapamycin alone, or combination of both drugs. The cellular effects of irinotecan and rapamycin were further characterized for HT-29 and HCT-116 colon cancer cells in vitro. RESULTS: In contrast to single-agent therapy, xenografted tumors treated with combination of irinotecan and rapamycin showed potent inhibition of the mammalian target of rapamycin/HIF-1alpha axis, which was accompanied by a dramatic reduction in tumor volume. In vitro experiments showed that exposure to low concentrations of the two drugs resulted in massive HT-29 cell death under hypoxic, but not normoxic, conditions, in full agreement with a cytotoxic effect mediated through HIF-1alpha rather than through induction of genotoxic lesions. HCT-116 cells were less sensitive to the combined treatment due to constitutive activation of phosphatidylinositol 3-kinase/Akt and Ras/mitogen-activated protein kinase pathways. CONCLUSION: These results identify HIF-1alpha as a promising target and provide a rationale for clinical trials of low-dose irinotecan and rapamycin combination toward metastatic colon cancer. PMID- 19190129 TI - Distinct BRAF (V600E) and KRAS mutations in high microsatellite instability sporadic colorectal cancer in African Americans. AB - PURPOSE: Colorectal cancer develops through genetic, epigenetic, and environmental events that result in uncontrolled cell proliferation. Colorectal cancer incidence and mortality is higher in African Americans (AA) than in the general population. Here, we carried out a molecular analysis of sporadic colorectal cancer tumors from AAs to investigate possible explanations for the observed disparities. EXPERIMENTAL DESIGN: A total of 222 AA colorectal cancer tumors were analyzed for microsatellite instability (MSI) for protein expression of two DNA mismatch repair genes, MLH1 and MSH2, by immunohistochemistry; for the methylation silencing of MLH1, p16, APC, and APC2 promoters by methylation specific PCR; and for point mutations in two oncogenes, KRAS and BRAF, by sequencing. RESULTS: In our sample, 19.8% of the AAs colorectal cancer tumors were MSI high (MSI-H) and did not associate with any of the clinicopathologic features, except tumor differentiation. Higher levels of inactive DNA mismatch repair proteins MLH1 (41%) and MSH2 (33%) were found by immunohistochemistry. Methylation-specific PCR analysis revealed a high level of methylation for MLH1 (66%), APC (53%), and APC2 (90%), but not for p16 (26%). BRAF mutations were only within the MSI-H tumors, whereas most (64%) of KRAS mutations were found within the non-MSI-H group. CONCLUSIONS: MLH1, MSH2, and BRAF alterations are significantly associated with MSI-H phenotype, unlike APC, APC2 and KRAS alterations. The prominent role of DNA mismatch repair gene suppression in MSI-H and a distinctive role of BRAF and KRAS mutations with respect to MSI status are supported by this study. PMID- 19190132 TI - The expression of three genes in primary non-small cell lung cancer is associated with metastatic spread to the brain. AB - PURPOSE: Brain metastases affect 25% of patients with non-small cell lung cancer (NSCLC). We hypothesized that the expression of genes in primary NSCLC tumors could predict brain metastasis and be used for identification of high-risk patients, who may benefit from prophylactic therapy. EXPERIMENTAL DESIGN: The expression of 12 genes was measured by real-time quantitative reverse transcriptase PCR in 142 frozen NSCLC tissue samples. Univariate and multivariate Cox regression analysis was used to analyze the correlation between gene expression and the occurrence of brain metastasis. Immunohistochemistry on independent samples was used to verify the findings. RESULTS: A score based on the expression levels of three genes, CDH2 (N-cadherin), KIFC1, and FALZ, was highly predictive of brain metastasis in early and advanced lung cancer. The probability of remaining brain metastasis-free at 2 years after diagnosis was 90.0+/-9.5% for patients with stage I/stage II tumors and low score compared with 62.7+/-12% for patients with high score (P<0.01). In patients with more advanced lung cancer, the brain metastasis-free survival at 24 months was 89% for patients with low score compared with only 37% in patients with high score (P<0.02). These results were confirmed by immunohistochemical detection of N-cadherin in independent cohort of primary NSCLC. CONCLUSIONS: The expression levels of three genes in primary NSCLC tumors may be used to identify patients at high risk for brain metastasis who may benefit from prophylactic therapy to the central nervous system. PMID- 19190134 TI - Communicable Disease and Health Protection Quarterly Review: September to December 2008. PMID- 19190135 TI - The quality of family planning services and client satisfaction in the public and private sectors in Kenya. AB - OBJECTIVE: To compare the quality of family planning services delivered at public and private facilities in Kenya. METHODS: Data from the 2004 Kenya Service Provision Assessment were analysed. The Kenya Service Provision Assessment is a representative sample of health facilities in the public and private sectors, and comprises data obtained from a facility inventory, service provider interviews, observations of client-provider interactions and exit interviews. Quality-of-care indicators are compared between the public and private sectors along three dimensions: structure, process and outcome. RESULTS: Private facilities were superior to public sector facilities in terms of physical infrastructure and the availability of services. Public sector facilities were more likely to have management systems in place. There was no difference between public and private providers in the technical quality of care provided. Private providers were better at managing interpersonal aspects of care. The higher level of client satisfaction at private facilities could not be explained by differences between public and private facilities in structural and process aspects of care. CONCLUSIONS: Formal private sector facilities providing family planning services exhibit greater readiness to provide services and greater attention to client needs than public sector facilities in Kenya. Consistent with this, client satisfaction is much higher at private facilities. Technical quality of care provided is similar in public and private facilities. PMID- 19190133 TI - Establishment and characterization of a model of acquired resistance to epidermal growth factor receptor targeting agents in human cancer cells. AB - PURPOSE: The epidermal growth factor receptor (EGFR) is recognized as a key mediator of proliferation and progression in many human tumors. A series of EGFR specific inhibitors have recently gained Food and Drug Administration approval in oncology. These strategies of EGFR inhibition have shown major tumor regressions in approximately 10% to 20% of advanced cancer patients. Many tumors, however, eventually manifest resistance to treatment. Efforts to better understand the underlying mechanisms of acquired resistance to EGFR inhibitors, and potential strategies to overcome resistance, are greatly needed. EXPERIMENTAL DESIGN: To develop cell lines with acquired resistance to EGFR inhibitors we utilized the human head and neck squamous cell carcinoma tumor cell line SCC-1. Cells were treated with increasing concentrations of cetuximab, gefitinib, or erlotinib, and characterized for the molecular changes in the EGFR inhibitor-resistant lines relative to the EGFR inhibitor-sensitive lines. RESULTS: EGFR inhibitor-resistant lines were able to maintain their resistant phenotype in both drug-free medium and in athymic nude mouse xenografts. In addition, EGFR inhibitor-resistant lines showed a markedly increased proliferation rate. EGFR inhibitor-resistant lines had elevated levels of phosphorylated EGFR, mitogen-activated protein kinase, AKT, and signal transducer and activator of transcription 3, which were associated with reduced apoptotic capacity. Subsequent in vivo experiments indicated enhanced angiogenic potential in EGFR inhibitor-resistant lines. Finally, EGFR inhibitor-resistant lines showed cross-resistance to ionizing radiation. CONCLUSIONS: We have developed EGFR inhibitor-resistant human head and neck squamous cell carcinoma cell lines. This model provides a valuable preclinical tool to investigate molecular mechanisms of acquired resistance to EGFR blockade. PMID- 19190137 TI - Physical activity and endogenous sex hormone levels in postmenopausal women: a cross-sectional study in the Prospect-EPIC Cohort. AB - BACKGROUND: The protective effect of physical activity on breast cancer risk might be mediated by sex hormone levels. In this study, we examined the association between usual physical activity and plasma levels of estrogens, androgens, dehydroepiandrosterone sulphate (DHEAS), and sex hormone binding globulin (SHBG) in postmenopausal women. METHODS: We conducted a cross-sectional study among 806 postmenopausal women participating in the Prospect-EPIC study. Usual physical activity was assessed using a short questionnaire and summarized into a simple, validated four-level index. This index combines occupational physical activity with time spent on cycling and sporting. Levels of estrone, estradiol, androstenedione, DHEAS, testosterone, and SHBG were measured in plasma. General linear models were used to examine the association between usual physical activity and sex hormone levels, adjusted for confounders. RESULTS: We observed an inverse association between physical activity and estradiol levels (free: inactive, 0.26 pg/mL; active, 0.23 pg/mL; P-trend = 0.045; total: inactive, 8.8 pg/mL; active, 8.0 pg/mL; P-trend = 0.08) and a positive association between physical activity and SHBG (inactive, 15.1 nmol/L; active, 19.3 nmol/L; P-trend = 0.05). These associations could largely be explained by the effects of sporting. We also observed a positive association between physical activity and DHEAS (inactive, 352.4 ng/mL; active, 460.3 ng/mL; P-trend = 0.01). CONCLUSIONS: Our results are in accordance with the hypothesis that usual physical activity decreases estradiol levels and increases SHBG. We also found that high levels of physical activity are associated with high levels of DHEAS. Furthermore, our results suggest that vigorous forms of physical activity influence sex hormone levels most. PMID- 19190136 TI - Association of thymidylate synthase gene with endometrial cancer risk in a Chinese population. AB - We comprehensively evaluated genetic variants in the thymidylate synthase (TYMS) gene in association with endometrial cancer risk in a population-based case control study of 1,199 incident endometrial cancer cases and 1,212 age frequency matched population controls. Exposure information was obtained via in-person interview, and DNA samples (blood or buccal cell) were collected. Genotyping of 11 haplotype-tagging single nucleotide polymorphisms (SNP) for the TYMS gene plus the 5-kb flanking regions was done for 1,028 cases and 1,003 controls by using the Affymetrix MegAllele Targeted Genotyping System. Of 11 haplotype-tagging SNPs identified, 7 that are located in flanking regions of the TYMS gene are also in the ENOSF1 (rTS) gene. The SNP rs3819102, located in the 3'-flanking region of the TYMS gene and in an intron of the ENOSF1 gene, was associated with risk of endometrial cancer. The odds ratio (95% confidence interval) for the CC genotype was 1.5 (1.0-2.2) compared with the TT genotype. Haplotype TTG in block 2 of the TYMS gene, which includes SNPs rs10502289, rs2298583, and rs2298581 (located in introns of the ENOSF1 gene), was associated with a marginally significant decrease in risk of endometrial cancer under the dominant model (odds ratio, 0.8; 95% confidence interval, 0.6-1.0). This study suggests that genetic polymorphisms in the TYMS or ENOSF1 genes may play a role in the development of endometrial cancer among Chinese women. PMID- 19190138 TI - Longitudinal biological exposure to carotenoids is associated with breast cancer free survival in the Women's Healthy Eating and Living Study. AB - In some cohort studies, a high-vegetable diet has been associated with greater likelihood of recurrence-free survival in women diagnosed with breast cancer. Carotenoids are obtained primarily from vegetables and fruit and they exhibit biological activities that may specifically reduce the progression of mammary carcinogenesis. The present analysis examines the relationship between plasma carotenoids at enrollment and 1, 2 or 3, 4, and 6 years and breast cancer-free survival in the Women's Healthy Eating and Living Study participants (N = 3,043), who had been diagnosed with early-stage breast cancer. The primary end point was time to a second breast cancer event (a recurrence or new primary breast cancer). An average carotenoid concentration over time was estimated for each participant as the average area under the plasma carotenoid curve formed by the plasma carotenoid concentrations at scheduled clinic visits. Multiple regression Cox proportional hazards analysis with adjustment for prognostic and other factors was used to examine the association between carotenoids and breast cancer-free survival. A total of 508 (16.7%) breast cancer events occurred over a median 7.12 years follow-up. Compared with the lowest tertile, the hazard ratio for the medium/high plasma carotenoid tertiles was 0.67 (95% confidence interval, 0.54 0.83) after adjustment. The interaction between the study group and tertile of average carotenoid concentration over time was not significant (P = 0.23). Higher biological exposure to carotenoids, when assessed over the time frame of the study, was associated with greater likelihood of breast cancer-free survival regardless of study group assignment. PMID- 19190139 TI - Is there a difference in the association between percent mammographic density and subtypes of breast cancer? Luminal A and triple-negative breast cancer. AB - BACKGROUND: Mammographic density is a potentially modifiable risk factor for breast cancer. To what extent mammographic density is a predictor for both hormone receptor-positive and hormone receptor-negative tumors is unclear. Even less is known about whether mammographic density predicts subtypes of breast cancer defined by expression status of the three receptors: estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor-2 (HER-2). METHODS: We estimated the association of percent mammographic density with subtypes of invasive breast cancer among 479 population-based female breast cancer patients and 376 control subjects ages 35 to 64 years. The expression status of ER, PR, and HER-2 was assessed using immunohistochemistry methods in a single laboratory. We considered ER+ or PR+ plus HER-2- tumors as luminal A breast cancer and ER-/PR-/HER-2- tumors as triple-negative breast cancer. We used unconditional logistic regression methods to estimate odd ratios (95% confidence intervals) for both case-control and case-case comparisons. RESULTS: Mammographic density was associated with increased risk of both invasive breast cancer subtypes, luminal A and triple-negative, in the case-control analysis. Results from case-case comparisons yielded no differences between the two subtypes among all women combined or in analyses done separately by race (White versus African American women) or menopausal status (premenopausal versus postmenopausal women; all P values > 0.05). CONCLUSIONS: Our results suggest that percent mammographic density is positively associated with both luminal A and triple-negative breast cancer. PMID- 19190140 TI - Influence of primary care use on population delivery of colorectal cancer screening. AB - OBJECTIVE: Colorectal cancer (CRC) screening is commonly initiated during primary care visits. Thus, at the population level, limited primary care attendance may constitute a substantial barrier to CRC screening uptake. Within a defined population, we quantified the percent of CRC screening underuse that is potentially explained by low use of primary care visits. METHODS: Among 48,712 adults ages 50 to 78 years eligible for CRC screening within a Washington state health plan, we estimated the degree to which a lack of CRC screening in 2002 to 2003 (fecal occult blood testing, sigmoidoscopy, or colonoscopy) was attributable to low primary care use, expressed as the population attributable risk percent (PAR%) associated with 0 to 3 primary care visits during the 2-year period. RESULTS: In analyses adjusted for age, comorbidity, nonprimary care visit use, and prior preventive service use, low primary care use in 2002 to 2003 was strongly associated with a lack of CRC screening among both women and men. However, a majority of unscreened women and men had > or =4 primary care visits. Thus, whether low primary care use was defined as 0, 0 to 1, 0 to 2, or 0 to 3 primary care visits, the PAR% associated with low primary care use was large in neither women (range, 3.0-6.8%) nor men (range: 5.6-11.5%). CONCLUSIONS: Health plan outreach efforts to encourage primary care attendance would be unlikely to substantially increase population uptake of CRC screening. In similar settings, resources might be more fruitfully devoted to the optimization of screening delivery during primary care visits that patients already attend. PMID- 19190141 TI - Antacid drug use and risk of esophageal and gastric adenocarcinomas in Los Angeles County. AB - OBJECTIVES: Concern has been expressed that antacid drugs increase the risk of esophageal and gastric adenocarcinomas. METHODS: This population-based case control study recruited patients with incident esophageal adenocarcinoma (n = 220), gastric cardiac adenocarcinoma (n = 277), or distal gastric adenocarcinoma (n = 441) diagnosed between 1992 and 1997, and 1,356 control participants in Los Angeles County. Unconditional polychotomous multivariable logistic regression analyses were done to evaluate the association between antacid drug use and these cancers. RESULTS: Among participants who took nonprescription acid neutralizing agents for >3 years, the odds ratio for esophageal adenocarcinoma was 6.32 compared with never users (95% confidence interval, 3.14-12.69; P(trend) < 0.01). Analyses stratified by history of physician diagnosed upper gastrointestinal (UGI) disorders revealed a greater increase in esophageal adenocarcinoma risk associated with nonprescription antacid use among persons with no UGI disorder than among those with an UGI disorder (homogeneity of trends P = 0.07). Regular use of nonprescription acid neutralizing agents was not associated with risk of adenocarcinomas of the gastric cardia or distal stomach. Regular use of prescription acid suppressive drugs was not associated with risk for any of these cancers. CONCLUSION: We found risk of esophageal adenocarcinoma was greater among long-term nonprescription acid neutralizing drugs in participants without physician-diagnosed UGI conditions than among those with these conditions; this may represent self medication for undiagnosed precursor conditions or it may be that nonprescription acid neutralizing drugs, taken without limitation on amount used when symptoms are most intense, may permit alkaline bile reflux into the lower esophagus, thereby increasing esophageal adenocarcinoma risk. PMID- 19190142 TI - Health risk appraisal models for mass screening for esophageal and pharyngeal cancer: an endoscopic follow-up study of cancer-free Japanese men. AB - PURPOSE: To assess the performance of our health risk appraisal (HRA) models for screening individuals at high risk of esophageal/pharyngeal squamous cell carcinoma (EPSCC). METHODS: Based on the results of our previous case-control study, we invented HRA models that enable screening for EPSCC cases in Japanese men with high sensitivity and specificity based on either their aldehyde dehydrogenase-2 genotype (HRA-G model) or alcohol flushing (HRA-F model) and drinking, smoking, and dietary habits. Follow-up endoscopy combined with esophageal iodine staining (median follow-up period: 5.0 years) was done on 404 Japanese men (50-78 years) who were registered as cancer-free controls in the previous study. RESULTS: The follow-up endoscopy resulted in a diagnosis of 6 esophageal SCC (T(is) in 5 and T(1) in 1), 1 hypopharyngeal SCC (T(2)), and 1 oropharyngeal SCC (T(2)). Seven and 6 of the 8 EPSCC cases were in the top 10% risk group at baseline according to the HRA-G and HRA-F models, respectively. The EPSCC detection rates per 100 person-years in the top 10% risk groups by the HRA G and HRA-F models were 4.38 (95% confidence interval, 1.76-9.01) and 3.48 (95% confidence interval, 1.28-7.58), respectively. Their age-adjusted relative risk was 95.1- and 26.3-fold, respectively (P < 0.0001), higher than in the bottom 90% risk groups. CONCLUSIONS: The high detection rates for EPSCC in the top 10% risk group of this preliminary follow-up study were in good agreement with those predicted by the HRA models and thus encouraged the screening based on our HRA models in larger populations of Japanese men. PMID- 19190143 TI - Dietary intake of omega-6 and omega-3 fatty acids and risk of colorectal cancer in a prospective cohort of U.S. men and women. AB - BACKGROUND: omega-6 and omega-3 polyunsaturated fatty acids intakes may play opposing roles in inflammation-driven colorectal carcinogenesis. We examined the relationship of these polyunsaturated fatty acids and the ratio of their intake with colorectal cancer risk in a large U.S. prospective cohort. DESIGN: Participants in the Cancer Prevention Study-II Nutrition Cohort completed a detailed questionnaire on diet, medical history, and lifestyle in 1999. Between 1999 and 2005, 869 incident colorectal cancer cases (452 men and 417 women) were identified among 99,080 participants (43,108 men and 55,972 women). Multivariate adjusted rate ratios were calculated using Cox proportional hazards models. RESULTS: The ratio of total omega-6 to total omega-3 intake was not associated with colorectal cancer risk in either sex. Contrary to our initial hypothesis, total omega-6 intake was inversely related to colorectal cancer risk in men [multivariate relative risk (95% confidence interval) for highest to lowest quartile, 0.81 (0.61-1.07); P(trend) = 0.07], and alpha-linolenic acid, the primary contributor to total omega-3 intake, was associated with increased risk in women for quartiles 2 through 4 versus the lowest quartile [relative risk (95% confidence interval), 1.50 (1.12-2.01), 1.40 (1.04-1.87), and 1.38 (1.02-1.85), respectively; P(trend) = 0.13]. In women, total omega-6 and marine omega-3 intake appeared to be associated with higher and lower risk, respectively, but associations were attenuated with adjustment for other risk factors. CONCLUSIONS: The ratio of omega-6 to omega-3 intake was not related to colorectal cancer risk in this cohort, which may be due to unexpected findings for the individual components. Differential associations by sex warrant further investigation. PMID- 19190144 TI - Iron overload is associated with hepatic oxidative damage to DNA in nonalcoholic steatohepatitis. AB - Several lines of evidence have suggested that oxidative stress plays an important role for the pathogenesis of nonalcoholic steatohepatitis (NASH). Therefore, by using immunohistochemical staining of liver biopsy samples, we measured hepatic 7,8-dihydro-8-oxo-2' deoxyguanosine (8-oxodG), a DNA base-modified product generated by hydroxyl radicals, of 38 NASH patients and compared with 24 simple steatosis and 10 healthy subjects. Relation of hepatic 8-oxodG with clinical, biochemical, and histologic variables and changes after iron reduction therapy (phlebotomy plus iron-restricted diet) were also examined. Hepatic 8-oxodG levels were significantly higher in NASH compared with simple steatosis (17.5 versus 2.0 8-oxodG-positive cells/10(5) microm(2); P < 0.0001). 8-oxodG was significantly related to iron overload condition, glucose-insulin metabolic abnormality, and severities of hepatic steatosis in NASH patients. Logistic regression analysis also showed that hepatic iron deposit and insulin resistance were independent variables associated with elevated hepatic 8-oxodG. After the iron reduction therapy, hepatic 8-oxodG levels were significantly decreased (from 20.7 to 13.8 positive cells/10(5) microm(2); P < 0.01) with concomitant reductions of serum transaminase levels in NASH patients. In conclusion, iron overload may play an important role in the pathogenesis of NASH by generating oxidative DNA damage and iron reduction therapy may reduce hepatocellular carcinoma incidence in patients with NASH. PMID- 19190145 TI - Nuclear overexpression of mitotic regulatory proteins in biliary tract cancer: correlation with clinicopathologic features and patient survival. AB - Mitosis dysregulation is common in cancers. This study explored the nuclear expression patterns and prognostic significance of mitotic regulatory proteins, including Aurora kinases, survivin, and p53, in biliary tract cancer (BTC). Archival tumor samples from 161 BTC patients who underwent surgery were tested for the expression of Aurora-A, Aurora-B, survivin, and p53 by immunohistochemistry. The potential endogeneity among the clinicopathologic variables and survival outcome was assessed by a generalized simultaneous equations model. Nuclear overexpression of Aurora-A, Aurora-B, survivin, and p53 was found in 79 (49.1%), 45 (28.0%), 55 (34.2%), and 55 (34.2%) patients, respectively. Intrahepatic cholangiocarcinoma, compared with the other two subtypes, had significantly higher proportions of nuclear overexpression of Aurora-B and survivin (37.8% and 47.3%, respectively). Simultaneous overexpression of Aurora-A and Aurora-B was correlated with that of p53. Overexpression of Aurora-B was also correlated with that of survivin and tumor grade. Our data indicate that simultaneous overexpression of Aurora-A and Aurora B, suggesting dysregulated mitosis is associated with worse survival in patients with BTC. Independent prognostic factors for poor overall survival included simultaneous overexpression of Aurora-A and Aurora-B (hazard ratio, 1.997; 95% confidence interval, 1.239-3.219; P = 0.0045) and tumor grade (hazard ratio, 2.117; 95% confidence interval, 1.339-3.348; P = 0.0013) assessed by a multivariate analysis stratified by American Joint Committee on Cancer stage and p53 overexpression. Endogeneity testing suggested that nuclear overexpression of p53 and tumor type may influence patient survival through their interactions with Aurora-A/Aurora-B expression and tumor grade. PMID- 19190147 TI - Incident invasive breast cancer, geographic location of residence, and reported average time spent outside. AB - There have been reports of greater breast cancer incidence and mortality at northern compared with southern latitudes postulated to be related to vitamin D exposure. Among 71,662 participants in the Women's Health Initiative Observational Study (WHIOS) free of cancer at baseline (1993-1998), associations were explored between incident invasive postmenopausal breast cancer (n = 2,535), over approximately 8.6 years follow-up, and the following: (a) region of residence at birth, age 15 years, age 35 years; (b) region of residence at WHIOS baseline; and (c) clinic center solar irradiance. Hazard ratios and 95% confidence intervals (CI) for breast cancer were estimated after adjustment for individual level confounders. There was no difference in breast cancer risk by region of earlier life, baseline residence, or solar irradiance measured in Langelys (gm-cal) per cm(2). There was an observed 15% decreased risk among women residing in areas of low versus high solar irradiance measured in Watts per m(2) (95% CI, 2-26%). However, the associated P(trend) of 0.20 was not significant. Conversely, women who reported spending on average <30 minutes versus >2 hours outside in daylight year round at WHIOS year 4 follow-up (n = 46,926), had a 20% (95% CI, 2-41%; P(trend) = 0.001) increased risk of breast cancer. In conclusion, region of residence and geographic solar irradiance are not consistently related to risk of breast cancer and may not be sufficient proxy measures for sunlight/vitamin D exposure. The observed association between time spent outside and breast cancer risk support the hypothesis that vitamin D may protect against breast cancer. PMID- 19190146 TI - Genetic variation in the androgen receptor gene and endometrial cancer risk. AB - Genetic variation in the androgen receptor (AR) gene may be associated with endometrial cancer risk based on the role of AR in regulating androgen levels. However, endometrial cancer studies reported inconsistent associations for a CAG repeat polymorphism in exon 1. Only one of these studies measured haplotype tagging single nucleotide polymorphisms (htSNP) in AR and found statistically nonsignificant, decreased associations with endometrial cancer risk. In a population-based case-control study of 497 cases and 1,024 controls, we examined the CAG repeat polymorphism and six htSNPs (rs962458, rs6152, rs1204038, rs2361634, rs1337080, and rs1337082), which cover an estimated 80% of the known common variation in AR among Caucasian populations. CAG repeat length was not significantly associated with endometrial cancer [odds ratio per unit increase in the average number of repeats, 1.02 (95% confidence interval, 0.97-1.08); P(trend) = 0.29]. Minor alleles in three correlated htSNPs (rs6152, rs1204038, and rs1337082; r(2) >0.6) were associated with increased risk for endometrial cancer. The strongest association was observed for rs6152, with the odds ratios (95% confidence interval) being 1.13 (0.89-1.44) for heterozygous and 2.40 (1.28 4.51) for homozygous minor genotypes (P(trend) = 0.02) compared with homozygous major allele genotype. However, these associations were not statistically significant after permutation adjustment for multiple comparisons (P(trend) > 0.09). Haplotype analyses did not reveal any additional associations with endometrial cancer. Results from our study, taken together with previously published studies, provide little evidence of a consistent association between common genetic variation in AR and endometrial cancer risk. PMID- 19190148 TI - Vacations to waterside locations result in nevus development in Colorado children. AB - Nevi are a main risk factor for malignant melanoma, and most nevi develop in childhood. This study examined the relationship between vacations and nevi in 681 White children born in 1998 who were lifetime residents of Colorado. Vacation histories were assessed through telephone interviews of parents, whereas nevus and phenotypic characteristics were assessed through skin exams at age 7. Multiple linear and logistic regression were used to assess the influence of vacations on counts of nevi <2 mm in size and the presence of any nevi > or = 2 mm after controlling for other variables. Each waterside vacation > or = 1 year before the exam at age 7 was found to be associated with a 5% increase in nevi <2 mm. Waterside vacations <1 year before the skin exam were not related to nevus count (<2 mm); regardless of timeframe, waterside vacations were not related to the presence of nevi > or = 2 mm. UV dose received on waterside vacations, number of days spent on waterside vacations, and nonwaterside vacations were not significantly related to nevi <2 or > or = 2 mm. These results suggest that there is a lag of at least 1 year in the development of new nevi after vacation sun exposure. It appears that a threshold dose of UV exposure is received quickly on each waterside vacation. Parents of young children should exercise caution in selection of vacation locations to reduce melanoma risk. PMID- 19190149 TI - Sperm-associated antigen 9, a novel biomarker for early detection of breast cancer. AB - To date, there have been no tumor biomarkers validated and incorporated into oncologic practice for the early diagnosis of breast cancer. Recently, we showed that sperm-associated antigen 9 (SPAG9), a member of cancer testis (CT) antigen family, is associated with ovarian carcinomas. In the present study, we investigated SPAG9 expression and humoral immune response in breast cancer. We further evaluated the diagnostic potential of autoantibodies to SPAG9 protein in various stages, grades, and histotypes of breast cancer. We analyzed the association of SPAG9 immunoreactivity score (IRS) with predicted risk of breast cancer recurrence over 10 years. Our reverse transcription-PCR and immunohistochemical analyses revealed SPAG9 expression in 88% breast cancer specimens independent of tumor stages and grades. Further, the humoral immune response against SPAG9 was detected in 80% breast cancer patients with SPAG9 expressing tumors. The linear regression modeling predicted a direct relationship between presence of lymphovascular invasion and high SPAG9 IRS, whereas the univariate and multivariate logistic regression models predicted a strong association of SPAG9 IRS with tumor grade. Further, our data indicated a significant higher trend of SPAG9 IRS with the predicted high risk of breast cancer recurrence. The present investigation reports for the first time SPAG9 expression and humoral immune response in early stages and low-grade breast cancer. Although our data indicated that autoantibodies against SPAG9 represent a promising approach for the development of biomarker, further large-scale validation studies are required to establish its potential use in early diagnosis and monitoring of breast cancer recurrence. PMID- 19190150 TI - Telomere length, current perceived stress, and urinary stress hormones in women. AB - Telomeres are repetitive DNA sequences that cap and protect the ends of chromosomes; critically short telomeres may lead to cellular senescence or carcinogenic transformation. Previous findings suggest a link between psychosocial stress, shorter telomeres, and chronic disease risk. This cross sectional study examined relative telomere length in relation to perceived stress and urinary stress hormones in a sample of participants (n = 647) in the National Institute of Environmental Health Sciences Sister Study, a cohort of women ages 35 to 74 years who have a sister with breast cancer. Average leukocyte telomere length was determined by quantitative PCR. Current stress was assessed using the Perceived Stress Scale and creatinine-adjusted neuroendocrine hormones in first morning urines. Linear regression models estimated differences in telomere length base pairs (bp) associated with stress measures adjusted for age, race, smoking, and obesity. Women with higher perceived stress had somewhat shorter telomeres [adjusted difference of -129bp for being at or above moderate stress levels; 95% confidence interval (CI), -292 to 33], but telomere length did not decrease monotonically with higher stress levels. Shorter telomeres were independently associated with increasing age (-27bp/year), obesity, and current smoking. Significant stress-related differences in telomere length were seen in women ages 55 years and older (-289bp; 95% CI, -519 to -59), those with recent major losses (-420bp; 95% CI, -814 to -27), and those with above-average urinary catecholamines (e.g., epinephrine: -484bp; 95% CI, -709 to -259). Although current perceived stress was only modestly associated with shorter telomeres in this broad sample of women, our findings suggest the effect of stress on telomere length may vary depending on neuroendocrine responsiveness, external stressors, and age. PMID- 19190151 TI - Physical activity among lung cancer survivors: changes across the cancer trajectory and associations with quality of life. AB - BACKGROUND: Regular physical activity may offer benefits to lung cancer survivors, many of whom experience quality-of-life (QOL) impairments. However, little is know about lung cancer survivors' engagement in physical activity across the cancer trajectory. The current study addressed this research gap and also examined the association between lung cancer survivors' physical activity and their QOL. METHODS: The study participants were 175 individuals who completed surgical treatment for early-stage non-small cell lung cancer 1 to 6 years previously. Participants completed a one-time survey regarding their current QOL and their engagement in physical activities currently, during the 6 months after treatment, and during the 6 months before diagnosis. RESULTS: Participants' reported engagement in both moderate and strenuous intensity activities was lower during the post-treatment period compared with before diagnosis and at the current time. Engagement in light intensity activities did not differ for the three time points. Almost two-thirds of participants did not engage in sufficient activity to meet national physical activity guidelines for any of the three time points. Lung cancer survivors who currently met physical activity guidelines reported better QOL in multiple domains than less active individuals. CONCLUSIONS: Engagement in physical activity among lung cancer survivors is particularly low during the early post-treatment period. Current engagement in physical activity is associated with better QOL. However, most lung cancer survivors do not meet physical activity guidelines and may benefit from interventions to promote engagement in regular physical activities. PMID- 19190152 TI - Social-cognitive and school factors in initiation of smoking among adolescents: a prospective cohort study. AB - AIMS: The aim of the present study was to examine the association between social cognitive factors, school factors, and smoking initiation among adolescents who had never smoked. METHODS: The study was based on longitudinal data on Danish adolescents attending randomly selected public schools. Adolescents enrolled in grade 7 (mean age, 13 years) who had never smoked (n = 912) were followed up for 6 months after baseline. Those who had still never smoked were followed up again 18 months after baseline, in grade 8 (n = 442). Social-cognitive factors were examined with five measures: self-efficacy, social influence (norms), social influence (behavior), social influence (pressure), and attitude. We used multilevel analyses to estimate the associations between social-cognitive factors at baseline and smoking initiation as well as the random effects of school, school class, and gender group in the school class. RESULTS: At the first follow up, we found significant associations between attitude, father's smoking, best friend's smoking, and smoking initiation. At the second follow-up, we found a significant association with pressure to smoke from friends. Of the school factors, gender group in the school class showed an effect at both first and second follow-up. CONCLUSION: Our results suggest that father's smoking, best friend's smoking, attitude, and pressure to smoke from friends affect smoking initiation. The results for school factors suggest an effect of classmates of the same gender, which has not previously been examined longitudinally. PMID- 19190153 TI - Smokers of menthol and nonmenthol cigarettes exhibit similar levels of biomarkers of smoke exposure. AB - There has been speculation that the addition of menthol to cigarettes may affect the manner in which cigarettes are smoked, potentially influencing smokers' exposures to smoke constituents that have been associated with smoking-related diseases. One hundred twelve male and female smokers participated in a parallel arm study to determine whether the ad libitum smoking of menthol cigarettes results in differences in smoke constituent exposure biomarkers in blood and urine relative to those smoking nonmenthol cigarettes having similar machine measured (Federal Trade Commission) yields of approximately 9 to 10 mg "tar." The study subjects were provided cigarettes of their preferred menthol or nonmenthol types prior to two 24-hour study intervals spaced one week apart. Carboxyhemoglobin levels were measured in blood samples drawn at midafternoon following the two 24-hour urine collection periods. Six urinary nicotine metabolites (nicotine, cotinine, trans-3'-hydroxycotinine and respective glucuronides) were determined as measures of nicotine intake, and urinary 4-(N nitrosomethylamino)-1-(3-pyridyl)-1-butanol (NNAL) and its glucuronide were determined to assess exposure to the tobacco-specific nitrosamine 4-(N nitrosomethylamino)-1-(3-pyridinyl)-1-butanone. Subjects' median blood carboxyhemoglobin values did not differ significantly between the cigarette types. Neither total urinary NNAL nor urinary nicotine equivalents exhibited statistically significant differences between the menthol and nonmenthol cigarette smokers. The present findings indicate that moderately heavy smokers of menthol and nonmenthol cigarettes of similar machine-generated smoke yield exhibit essentially identical levels of biomarkers of smoke constituent exposure. These results are consistent with the substantial majority of epidemiology studies to date that suggest the risks attending the smoking of menthol and nonmenthol cigarettes are similar. PMID- 19190154 TI - Reproductive and hormonal factors, and ovarian cancer risk for BRCA1 and BRCA2 mutation carriers: results from the International BRCA1/2 Carrier Cohort Study. AB - BACKGROUND: Several reproductive and hormonal factors are known to be associated with ovarian cancer risk in the general population, including parity and oral contraceptive (OC) use. However, their effect on ovarian cancer risk for BRCA1 and BRCA2 mutation carriers has only been investigated in a small number of studies. METHODS: We used data on 2,281 BRCA1 carriers and 1,038 BRCA2 carriers from the International BRCA1/2 Carrier Cohort Study to evaluate the effect of reproductive and hormonal factors on ovarian cancer risk for mutation carriers. Data were analyzed within a weighted Cox proportional hazards framework. RESULTS: There were no significant differences in the risk of ovarian cancer between parous and nulliparous carriers. For parous BRCA1 mutation carriers, the risk of ovarian cancer was reduced with each additional full-term pregnancy (P trend = 0.002). BRCA1 carriers who had ever used OC were at a significantly reduced risk of developing ovarian cancer (hazard ratio, 0.52; 95% confidence intervals, 0.37 0.73; P = 0.0002) and increasing duration of OC use was associated with a reduced ovarian cancer risk (P trend = 0.0004). The protective effect of OC use for BRCA1 mutation carriers seemed to be greater among more recent users. Tubal ligation was associated with a reduced risk of ovarian cancer for BRCA1 carriers (hazard ratio, 0.42; 95% confidence intervals, 0.22-0.80; P = 0.008). The number of ovarian cancer cases in BRCA2 mutation carriers was too small to draw definitive conclusions. CONCLUSIONS: The results provide further confirmation that OC use, number of full-term pregnancies, and tubal ligation are associated with ovarian cancer risk in BRCA1 carriers to a similar relative extent as in the general population. PMID- 19190155 TI - Serum levels of soluble mesothelin-related peptides in malignant and nonmalignant asbestos-related pleural disease: relation with past asbestos exposure. AB - BACKGROUND: Malignant pleural mesothelioma (MPM) results from malignant transformation of mesothelial cells. Past asbestos exposure represents a major risk factor for MPM and other benign pleural disease. Soluble mesothelin-related peptides (SMRP) have been regarded as a promising serum biomarker for MPM. The aim of this study was to investigate serum levels of SMRP in malignant and nonmalignant asbestos-related pleural disease. PATIENTS: Four groups of patients were investigated: group 1 composed of 48 healthy subjects, group 2 composed of 177 patients with previous asbestos exposure and no pleural disease, group 3 composed of 36 patients with MPM, and group 4 composed of 101 patients with previous asbestos exposure and benign pleural disease. Serum SMRP levels were determined by ELISA. RESULTS: Serum SMRP levels were significantly higher among group 3 than the other three groups. There were no differences in SMRP concentrations between groups 2 and 4. Subjects exposed to asbestos had higher SMRP concentrations than normal control subjects regardless of the presence of pleural disease. The area under the receiver operating characteristic curve for SMRP values was 0.75 (95% confidence interval, 0.68-0.83). The SMRP level at 0.55 nmol/L/L was determined as the most optimal cutoff value with resulting sensitivity and specificity of 72% and 72% for the diagnosis of MPM. CONCLUSIONS: These data attest to good diagnostic sensitivity and specificity of SMRP for the diagnosis of malignant mesothelioma. We have also shown that serum SMRP levels might serve as a marker of asbestos exposure. PMID- 19190156 TI - The impact of human papillomavirus information on perceived risk of cervical cancer. AB - There is a need to develop public education about the link between human papillomavirus (HPV) and cervical cancer. Explaining that a sexually transmitted virus causes cervical cancer may affect perceived risk of cervical cancer. We hypothesized that presenting HPV information would have differential effects depending on age and screening attendance. Data were collected during face-to face interviews with a sample of British women age 16 to 75 years who had not heard of HPV before (n = 965). A repeated measures design was used, assessing perceived risk of cervical cancer before and after providing information about HPV. Perceived risk was assessed using a comparative risk measure with a five point response scale. Preinformation, the mean perceived risk score was 2.64 (SE, 0.03). Overall, presentation of HPV information did not have an effect on perceived risk of cervical cancer [chi(2)(1) = .72; P = 0.396], but as expected, there was a significant time by age interaction for the change in perceived risk [chi(2)(5) = 33.56; P < 0.001], which increased in the youngest age group (16-25 years) and decreased in the oldest age group (65-75 years). In a separate analysis with women in the screening age range (25-64 years; n = 709), there was a significant time by screening attendance interaction [chi(2)(1) = 5.25; P = 0.022], with an increase in perceived risk among women who did not regularly attend screening. This is the first study to examine the effect of HPV information on perceived risk across different population groups. Interventions to increase awareness of HPV could benefit from tailoring information to prescreening age, screening age, and postscreening age women. PMID- 19190157 TI - Race/ethnicity, physical activity, and quality of life in breast cancer survivors. AB - PURPOSE: To examine associations between recreational physical activity and quality of life (QOL) in a multiethnic cohort of breast cancer survivors, specifically testing whether associations are consistent across racial/ethnic groups after accounting for relevant medical and demographic factors that might explain disparities in QOL outcomes. METHODS: Data were collected from a population-based cohort of non-Hispanic White (n = 448), Black (n = 197), and Hispanic (n = 84) breast cancer survivors (stage 0-IIIa) in the Health, Eating, Activity, and Lifestyle Study. Physical activity was assessed approximately 2.5 years after breast cancer diagnosis, with QOL assessed on average 6 to 12 months later. We used structural equation modeling to examine relationships between meeting recommended levels of physical activity and QOL, stratifying by race/ethnicity and adjusting for other demographic, comorbidity, and treatment effects. RESULTS: Structural equation modeling indicated that meeting recommended levels of physical activity had significant positive associations with QOL for Black and non-Hispanic White women (P < 0.05). Fewer Black women reported meeting recommended physical activity levels (P < 0.001), but meeting recommendations was associated with better QOL. Post hoc tests showed that meeting physical activity recommendations was specifically associated with better vitality, social functioning, emotional roles, and global QOL (P < 0.05 for all). CONCLUSIONS: These results suggest that meeting recommended levels of physical activity is associated with better QOL in non-Hispanic White and Black breast cancer survivors. Findings may help support future interventions among breast cancer survivors and promote supportive care that includes physical activity, although more research is needed to determine these relationships among Hispanic and other ethnic minority women. PMID- 19190158 TI - Interaction between tobacco and alcohol use and the risk of head and neck cancer: pooled analysis in the International Head and Neck Cancer Epidemiology Consortium. AB - BACKGROUND: The magnitude of risk conferred by the interaction between tobacco and alcohol use on the risk of head and neck cancers is not clear because studies have used various methods to quantify the excess head and neck cancer burden. METHODS: We analyzed individual-level pooled data from 17 European and American case-control studies (11,221 cases and 16,168 controls) participating in the International Head and Neck Cancer Epidemiology consortium. We estimated the multiplicative interaction parameter (psi) and population attributable risks (PAR). RESULTS: A greater than multiplicative joint effect between ever tobacco and alcohol use was observed for head and neck cancer risk (psi = 2.15; 95% confidence interval, 1.53-3.04). The PAR for tobacco or alcohol was 72% (95% confidence interval, 61-79%) for head and neck cancer, of which 4% was due to alcohol alone, 33% was due to tobacco alone, and 35% was due to tobacco and alcohol combined. The total PAR differed by subsite (64% for oral cavity cancer, 72% for pharyngeal cancer, 89% for laryngeal cancer), by sex (74% for men, 57% for women), by age (33% for cases <45 years, 73% for cases >60 years), and by region (84% in Europe, 51% in North America, 83% in Latin America). CONCLUSIONS: Our results confirm that the joint effect between tobacco and alcohol use is greater than multiplicative on head and neck cancer risk. However, a substantial proportion of head and neck cancers cannot be attributed to tobacco or alcohol use, particularly for oral cavity cancer and for head and neck cancer among women and among young-onset cases. PMID- 19190159 TI - Serum high-density lipoprotein cholesterol and breast cancer risk by menopausal status, body mass index, and hormonal receptor in Korea. AB - High-density lipoprotein cholesterol (HDL-C) has been suggested to be associated with breast cancer. However, the roles of HDL-C and hypertriglyceridemia on breast cancer still have been controversial. The goal of this study was to investigate the association between HDL-C with breast cancer risk, stratifying by menopausal status, and body mass index. The hormonal receptor status of breast has been proposed to modify the effect of HDL-C on breast cancer. Multicenter hospital-based case-control study was conducted from November 2004 to December 2005 in Korea. After one to two individual matchings by age (+/-5 years) and menopausal status, 690 cases and 1,380 controls were included in the analysis. Odds ratios (OR) and 95% confidence intervals (95% CI) were estimated by conditional, unconditional, and multinomial logistic regressions. Protective effect of HDL-C on breast cancer was only observed among premenopausal women with an OR (95% CI) of 0.49 (0.33-0.72) for HDL-C > or = 60 versus <50 mg/dL (P(trend) < 0.01). Only nonobese premenopausal women had a significant decreased risk (OR, 0.34; 95% CI, 0.22-0.53). OR (95% CI) of low HDL-C (<50 mg/dL) and high triglyceride (TG; > or = 150 mg/dL) category was 2.20 (1.32-3.67) on estrogen receptor-negative and progesterone receptor-negative breast cancer compared with high HDL-C (> or = 50 mg/dL) and low TG (<150 mg/dL) category. This study suggests that higher level of HDL-C may reduce breast cancer risk among premenopausal women. Estrogen receptor-negative and progesterone receptor negative breast cancer was associated with dyslipidemia, which implicates that association among HDL-C, TG, and breast cancer may be modified by receptor status. PMID- 19190161 TI - Disparities in how parents are learning about the human papillomavirus vaccine. AB - BACKGROUND: Differential access to basic health information may contribute to persistent cervical cancer disparities. We examined whether human papillomavirus (HPV) vaccine awareness, HPV knowledge, and use of information sources about the vaccine differ by sociodemographic characteristics associated with cervical cancer. METHODS: Study participants (n = 889) were caregivers of adolescent girls ages 10 to 18 years living in southeastern North Carolina. Analyses simultaneously controlled for caregivers' gender, race, age, education, income, and rural residence. RESULTS: Although most caregivers were aware of HPV (83%) and the HPV vaccine (82%), awareness differed by gender, race, education, and income. The largest differences were for race, with 87% of Whites versus 68% of African Americans having heard of the vaccine (P < 0.001). Caregivers correctly answered an average of 69% of questions on HPV, with differences by race and education. Most respondents heard of the HPV vaccine through drug company advertisements (83%) or broadcast media coverage (69%). African Americans were less likely than Whites to have heard about the vaccine from advertisements but more likely from a broadcast source (P < 0.05). Health care providers (88%) and the internet (65%) were the most favored sources for future information about the vaccine. Vaccine uptake was associated with awareness, knowledge, and media use. DISCUSSION: Whereas drug company advertisements seem to play a central role in high HPV vaccine awareness, doctors and the internet are the preferred future "go to" sources for seeking out information. Communication-based interventions for caregivers from cervical cancer risk groups, especially African Americans, may need to use different communication channels and content. PMID- 19190162 TI - Risk factors for pancreatic cancer mortality: extended follow-up of the original Whitehall Study. AB - Given the well-established links between diabetes and elevated rates of pancreatic cancer, there are reasons to anticipate that other markers of metabolic abnormality (increased body mass index, plasma cholesterol, and blood pressure) and their correlates (physical activity and socioeconomic status) may also confer increased risk. However, to date, the results of a series of population-based cohort studies are inconclusive. We examined these associations in the original Whitehall cohort study of 17,898 men. A maximum of 38 years of follow-up gave rise to 163 deaths due to carcinoma of the pancreas. Although Poisson regression analyses confirmed established risk factor-disease associations for increasing age, smoking, and type II diabetes, there was essentially no evidence that body mass index (rate ratio, 1.01; 95% confidence interval per 1 SD increase, 0.86-1.18), plasma cholesterol (0.91; 0.78-1.07), diastolic blood pressure (0.93; 0.78-1.09), systolic blood pressure (0.98; 0.83 1.15), physical activity (sedentary versus high: 1.37; 0.89-2.12), or socioeconomic status [clerical (low) versus professional/executive, 0.95; 0.59 1.51] offered any predictive value for pancreatic cancer mortality. These results were unchanged following control for a range of covariates. PMID- 19190163 TI - Antidepressant medications and change in mammographic density in postmenopausal women. AB - BACKGROUND: Several studies suggest antidepressant medications can increase prolactin levels. Some, but not all, studies suggest prolactin levels are positively associated with mammographic breast density, an established risk factor for breast cancer. METHODS: We evaluated 29,284 women with two routine screening mammograms 9 to 28 months apart between 1996 to 2006 to examine whether antidepressant use was associated with changes in mammographic breast density. Mammographic breast density was assigned by radiologists and coded according the Breast Imaging Reporting and Data System classification. Exposure to antidepressants was ascertained based on electronic pharmacy dispensing data, including dispensings from claims data. We used polytomous logistic regression to estimate the odds of an increase or decrease in density categories between mammograms associated with antidepressant initiation, continuation, and discontinuation compared with nonusers of any antidepressants. RESULTS: Initiation, continuation, and discontinuation of antidepressant medications were not associated with changes in mammographic density. CONCLUSIONS: The lack of association between antidepressant use and breast density is consistent with recent studies that do not suggest an association between antidepressant use and breast cancer risk. PMID- 19190164 TI - Bladder cancer-associated gene expression signatures identified by profiling of exfoliated urothelia. AB - Bladder cancer is the fifth most commonly diagnosed malignancy in the United States and one of the most prevalent worldwide. It harbors a probability of recurrence of >50%; thus, rigorous, long-term surveillance of patients is advocated. Flexible cystoscopy coupled with voided urine cytology is the primary diagnostic approach, but cystoscopy is an uncomfortable, invasive procedure and the sensitivity of voided urine cytology is poor in all but high-grade tumors. Thus, improvements in noninvasive urinalysis assessment strategies would benefit patients. We applied gene expression microarray analysis to exfoliated urothelia recovered from bladder washes obtained prospectively from 46 patients with subsequently confirmed presence or absence of bladder cancer. Data from microarrays containing 56,000 targets was subjected to a panel of statistical analyses to identify bladder cancer-associated gene signatures. Hierarchical clustering and supervised learning algorithms were used to classify samples on the basis of tumor burden. A differentially expressed geneset of 319 gene probes was associated with the presence of bladder cancer (P < 0.01), and visualization of protein interaction networks revealed vascular endothelial growth factor and angiotensinogen as pivotal factors in tumor cells. Supervised machine learning and a cross-validation approach were used to build a 14-gene molecular classifier that was able to classify patients with and without bladder cancer with an overall accuracy of 76%. Our results show that it is possible to achieve the detection of bladder cancer using molecular signatures present in exfoliated tumor urothelia. Further investigation and validation of the cancer-associated profiles may reveal important biomarkers for the noninvasive detection and surveillance of bladder cancer. PMID- 19190165 TI - Rad50 c.687delT does not contribute significantly to familial breast cancer in a French population. AB - Mutations in DNA repair genes are known for their association with hereditary breast cancer. BRCA1 and BRCA2 are the major genes for high-penetrance familial breast and ovarian cancer, whereas mutations in ATM or Chek2 confer more modest cancer risk. Additional genes involved in DNA double-strand break repair have more recently been associated with breast cancer risk: heterozygosity for deleterious mutations in components of the Rad50-Mre11-Nbs1 complex seems to predispose to breast cancer. In particular, the c.687delT mutation in Rad50 conferred an odds ratio of 4.3 for the risk of breast cancer in a study of Finnish breast cancer families. To explore the contribution of this mutation to breast cancer in French families for which no BRCA mutation could be found, we analyzed the relevant exon in 618 familial breast cancer cases and 513 controls with no personal or familial history of breast cancer. Rad50 was analyzed in its entirety for 231 familial cases, with no clearly deleterious mutations detected. These data together suggest that although founder mutations may make Rad50 a significant breast cancer risk factor in certain populations, it is not a factor in others. PMID- 19190166 TI - Prospective evaluation of hepatitis B 1762(T)/1764(A) mutations on hepatocellular carcinoma development in Shanghai, China. AB - Chronic infection with the hepatitis B virus (HBV) is the most important risk factor for hepatocellular carcinoma (HCC). However, determinants of HCC risk in infected individuals are not well understood. We prospectively evaluated the association between acquired HBV 1762(T)/1764(A) double mutations and HCC risk among 49 incident HCC cases and 97 controls with seropositive hepatitis B surface antigen at baseline from a cohort of 18,244 men in Shanghai, China, enrolled during 1986 to 1989. Compared with HBV carriers without the mutations, chronic HBV carriers with the HBV 1762(T)/1764(A) double mutations experienced an elevated risk of HCC (odds ratio, 2.47; 95% confidence interval, 1.04-5.85; P = 0.04). Risk increased with increasing copies of the double mutations; men with > or =500 copies/microL serum had an odds ratio of 14.57 (95% confidence interval, 2.41-87.98) relative to those without the double mutations (P(trend) = 0.004). Thus, the HBV 1762(T)/1764(A) double mutation is a codeterminant of HCC risk for people chronically infected with HBV. PMID- 19190167 TI - A two-stage case-control study of EGFR polymorphisms and breast cancer risk. PMID- 19190168 TI - Use of molecular testing to identify a cluster of patients with polycythemia vera in eastern Pennsylvania. AB - BACKGROUND: The role of the environment in the origin of polycythemia vera has not been well documented. Recently, molecular diagnostic tools have been developed to facilitate the diagnosis of polycythemia vera. A cluster of patients with polycythemia vera was suspected in three countries in eastern Pennsylvania where there have long been a concern about environment hazards. METHODS: Rigorous clinical criteria and JAK2 617V>F testing were used to confirm the diagnosis of polycythemia vera in patients in this area. Participants included cases of polycythemia vera from the 2001 to 2005 state cancer registry as well as self- and physician-referred cases. FINDING: A diagnosis of polycythemia vera was confirmed in 53% of 62 participants using WHO criteria, which includes JAK2 617V>F testing. A statistically significant cluster of cases (P < 0.001) was identified where the incidence of polycythemia vera was 4.3 times that of the rest of the study area. The area of the cluster contained numerous sources of hazardous material including waste-coal power plants and U.S. Environmental Protection Agency Superfund sites. INTERPRETATION: The diagnosis of polycythemia vera based solely on clinical criteria is frequently erroneous, suggesting that our prior knowledge of the epidemiology of this disease might be inaccurate. The JAK2 617V>F mutational analysis provides diagnostic clarity and permitted the confirmation of a cluster of polycythemia vera cases not identified by traditional clinical and pathologic diagnostic criteria. The close proximity of this cluster to known areas of hazardous material exposure raises concern that such environmental factors might play a role in the origin of polycythemia vera. PMID- 19190169 TI - Immunosuppression and other risk factors for lip cancer after kidney transplantation. AB - Incidence of lip cancer is markedly increased after kidney transplantation. Immunosuppression and other risk factors for lip cancer were investigated in a population-based, nationwide cohort of 8,162 kidney transplant recipients registered on the Australia and New Zealand Dialysis and Transplant Registry (1982-2003). Lip cancer diagnoses were ascertained using probabilistic data linkage with the Australian National Cancer Statistics Clearing House. Standardized incidence ratios were used to compare lip cancer risk by subsite of lip and during periods of transplant function and failure. Risk factors during the first functioning transplant were examined using multivariate Poisson regression. Lip cancer was diagnosed in 203 patients. All cases were of squamous cell origin and mostly (77%) affected the lower vermillion. Cases occurred predominantly during periods of transplant function, with incidence decreasing to pretransplantation level on transplant failure and cessation of immunosuppression. During transplant function, cancer of the lower vermillion was associated with increasing year of age [incidence rate ratio (IRR), 1.03; 95% confidence interval (95% CI), 1.02-1.05], greater time since transplantation (P < 0.001), smoking (IRR, 2.13; 95% CI, 1.12-4.07), and current use of azathioprine (IRR, 2.67; 95% CI, 1.39-5.15) or cyclosporine (IRR, 1.63; 95% CI, 1.00-2.65). Female sex (IRR, 0.29; 95% CI, 0.18-0.46) and non-Australian/New Zealand country of birth (P = 0.006), surrogate indices of reduced exposure to solar UV radiation, were significantly protective. Lip cancer after transplantation is strongly related to the current receipt of immunosuppression. During transplant function, lip cancer risk is associated with the duration of immunosuppression, receipt of specific immunosuppressive agents, and UV exposure. PMID- 19190170 TI - A prospective study of total and ionized serum calcium and fatal prostate cancer. AB - We recently reported a significant positive association in the National Health and Nutrition Examination Survey between high levels of total calcium in serum, measured prospectively, and risk of fatal prostate cancer. To confirm this, we examined associations between total and ionized serum calcium and prostate cancer mortality in an independent cohort, the Third National Health and Nutrition Examination Survey. Twenty-five prostate cancer deaths occurred over 56,625 person-years of follow-up. Compared with men in the lowest tertile of total serum calcium, the multivariate-adjusted relative risk for death from prostate cancer for men in the highest tertile was 2.07 (95% confidence interval, 1.06-4.04). For ionized serum calcium, the physiologically active fraction of total serum calcium, the relative risk for men in the highest tertile was 3.18 (95% confidence interval, 1.09-9.28). These findings support the hypothesis that serum calcium is a prospective biomarker of fatal prostate cancer. PMID- 19190171 TI - Assessment of lifetime cumulative sun exposure using a self-administered questionnaire: reliability of two approaches. AB - Few studies have evaluated the reliability of lifetime sun exposure estimated from inquiring about the number of hours people spent outdoors in a given period on a typical weekday or weekend day (the time-based approach). Some investigations have suggested that women have a particularly difficult task in estimating time outdoors in adulthood due to their family and occupational roles. We hypothesized that people might gain additional memory cues and estimate lifetime hours spent outdoors more reliably if asked about time spent outdoors according to specific activities (an activity-based approach). Using self administered, mailed questionnaires, test-retest responses to time-based and to activity-based approaches were evaluated in 124 volunteer radiologic technologist participants from the United States: 64 females and 60 males 48 to 80 years of age. Intraclass correlation coefficients (ICC) were used to evaluate the test retest reliability of average number of hours spent outdoors in the summer estimated for each approach. We tested the differences between the two ICCs, corresponding to each approach, using a t test with the variance of the difference estimated by the jackknife method. During childhood and adolescence, the two approaches gave similar ICCs for average numbers of hours spent outdoors in the summer. By contrast, compared with the time-based approach, the activity based approach showed significantly higher ICCs during adult ages (0.69 versus 0.43, P = 0.003) and over the lifetime (0.69 versus 0.52, P = 0.05); the higher ICCs for the activity-based questionnaire were primarily derived from the results for females. Research is needed to further improve the activity-based questionnaire approach for long-term sun exposure assessment. PMID- 19190172 TI - In vivo role of cytochrome P450 2E1 and glutathione-S-transferase activity for acrylamide toxicokinetics in humans. AB - Acrylamide, a potential food carcinogen in humans, is biotransformed to the epoxide glycidamide in vivo. Both acrylamide and glycidamide are conjugated with glutathione, possibly via glutathione-S-transferases (GST), and bind covalently to proteins and nucleic acids. We investigated acrylamide toxicokinetics in 16 healthy volunteers in a four-period change-over trial and evaluated the respective role of cytochrome P450 2E1 (CYP2E1) and GSTs. Participants ingested self-prepared potato chips containing acrylamide (1 mg) without comedication, after CYP2E1 inhibition (500 mg disulfiram, single dose) or induction (48 g/d ethanol for 1 week), and were phenotyped for CYP2E1 with chlorzoxazone (250 mg, single dose). Unchanged acrylamide and the mercapturic acids N-acetyl-S-(2 carbamoylethyl)-cysteine (AAMA) and N-acetyl-S-(2-hydroxy-2-carbamoylethyl) cysteine (GAMA) accounted for urinary excretion [geometric mean (percent coefficient of variation)] of 2.9% (42), 65% (23), and 1.7% (65) of the acrylamide dose in the reference period. Hemoglobin adducts increased clearly following the acrylamide test-meal. The cumulative amounts of acrylamide, AAMA, and GAMA excreted and increases in AA adducts changed significantly during CYP2E1 blockade [point estimate (90% confidence interval)] to the 1.34-fold (1.14-1.58), 1.18-fold (1.02-1.36), 0.44-fold (0.31-0.61), and 1.08-fold (1.02-1.15) of the reference period, respectively, but were not changed significantly during moderate CYP2E1 induction. Individual baseline CYP2E1 activity, CYP2E1*6, GSTP1 313A>G and 341T>C single nucleotide polymorphisms, and GSTM1-and GSTT1-null genotypes had no major effect on acrylamide disposition. The changes in acrylamide toxicokinetics upon CYP2E1 blockade provide evidence that CYP2E1 is a major but not the only enzyme mediating acrylamide epoxidation in vivo to glycidamide in humans. No obvious genetic risks or protective factors in xenobiotic-metabolizing enzymes could be determined for exposed subjects. PMID- 19190173 TI - Prior victimization and sexual and contraceptive self-efficacy among adolescent females under Child Protective Services care. AB - Adolescent females under Child Protective Services care in Quebec, Canada (n = 328) completed a questionnaire designed to explore associations between prior victimization (childhood sexual abuse and four forms of dating violence) and four dimensions of sexual and contraceptive self-efficacy. Five MANCOVAs were performed. In each model, a victimization index served as the independent variable, four Contraceptive Self-Efficacy subscales as dependant variables, and age, age at first sexual intercourse, and lifetime number of sexual partners as covariates. Childhood sexual abuse was negatively associated with adolescent's perceived ability to communicate about her sexuality and contraceptive practices. Sexual coercion in dating relationships was negatively associated with the adolescent's perceived ability to communicate about her sexuality and contraception, her perceived control over her sexual activity, and her perceived control over her use of contraceptives in passionate situations. Sexual victimization was found to impair important aspects of adolescent females' sexual and contraceptive self-efficacy. PMID- 19190175 TI - On receiving the diagnosis of cancer: the adolescent perspective. AB - The adolescent life stage encompasses unique developmental challenges. Little is known about the experience of receiving a cancer diagnosis during this already demanding life stage. The aim of this study is to explore the lived experience of being diagnosed with cancer from the perspective of the adolescent. A phenomenological approach is used to perform semistructured interviews with 10 adolescents who have been diagnosed with cancer within the previous 4 to 6 months. Colaizzi's method of analysis is used to allow themes to emerge from these data. Six themes capture the essence of being diagnosed with cancer from the perspective of the adolescent: (1) the stunning loss of normalcy, (2) gaining information, (3) the importance of friends and their reactions, (4) getting used to it, (5) giving back, and (6) family support. The results of this study provide an emerging understanding of the unique perspective of the adolescent with cancer. Understanding this perspective is critical to meet the needs of this population. PMID- 19190174 TI - Mitogen-activated protein kinase pathways contribute to hypercontractility and increased Ca2+ sensitization in murine experimental colitis. AB - Inflammatory bowel disease (IBD) is associated with intestinal smooth muscle dysfunction. Many smooth muscle contractile events are associated with alterations in Ca(2+)-sensitizing pathways. The aim of the present study was to assess the effect of colitis on Ca(2+) sensitization and the signaling pathways responsible for contractile dysfunction in murine experimental colitis. Colitis was induced in BALB/c mice by providing 5% dextran sulfate sodium (DSS) in drinking water for 7 days. Contractile responses of colonic circular smooth muscle strips to 118 mM K(+) and carbachol (CCh) were assessed. DSS induced a T(H)2 colitis [increased interleukin (IL)-4 and IL-6] with no changes in T(H)1 cytokines. Animals exposed to DSS had increased CCh-induced contraction (3.5 fold) and CCh-induced Ca(2+)-sensitization (2.2-fold) responses in intact and alpha-toxin permeabilized colonic smooth muscle, respectively. The contributions of extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein kinase (MAPK) to CCh-induced contractions were significantly increased during colitis. Ca(2+)-independent contraction induced by microcystin was potentiated (1.5-fold) in mice with colitis. ERK and p38MAPK (but not Rho-associated kinase) contributed to this potentiation. ERK1/2 and p38MAPK expression were increased in the muscularis propria of colonic tissue from both DSS-treated mice and patients with IBD (ulcerative colitis >> Crohn's disease). Murine T(H)2 colitis resulted in colonic smooth muscle hypercontractility with increased Ca(2+) sensitization. Both ERK and p38MAPK pathways contributed to this contractile dysfunction, and expression of these molecules was altered in patients with IBD. PMID- 19190176 TI - Ethical considerations in pediatric oncology phase I clinical trials according to The Belmont Report. AB - Whether to suggest enrollment of pediatric oncology patients with advanced or refractory disease into a Phase I clinical trial may present a significant ethical dilemma for health care professionals. Phase I trials are experimental and unpredictable by nature, yet health care professionals must ensure the trial's therapeutic intent as well as address the many vulnerabilities of the child with terminal cancer. After reviewing the role and phases of clinical research in pediatric oncology, this article discusses ethical considerations in Phase I clinical trials according to The Belmont Report: Ethical Principles and Guidelines for the Protection of Human Subjects in Research and discusses specific applications of these key ethical principles. PMID- 19190177 TI - Parental fears following their child's brain tumor diagnosis and treatment. AB - The objective of this study is to portray the illness-related threats experienced by parents of children after the diagnosis of central nervous system (CNS) tumor. Parents were asked to rate the extent to which they experienced a set of specific fears related to their child's brain tumor and its treatment. Outcomes for parents of CNS tumor patients (n = 82) were compared with those of reference parents of patients treated for acute lymphoblastic leukemia (n = 208). The fears about an illness recurrence and the late effects of treatment were most prominent among parents of CNS tumor patients. For 7 out of 11 kinds of fear, parents of CNS tumor patients expressed a stronger fear than the reference group. More than a quarter of the parents of children treated for CNS tumors feared a complete decline of the child. Parents of CNS tumor patients experience relatively heightened cancer related fears in several domains. The fear of devastating consequences felt by one fourth of parents signals the need of individualized psychological support and information at diagnosis and follow-up to facilitate parental coping with the posttreatment situation. PMID- 19190178 TI - Differential NF-kappaB pathways induction by Lactobacillus plantarum in the duodenum of healthy humans correlating with immune tolerance. AB - How do we acquire immune tolerance against food microorganisms and commensal bacteria that constitute the intestinal microbiota? We investigated this by stimulating the immune system of adults with commensal Lactobacillus plantarum bacteria. We studied the in vivo human responses to L. plantarum in a randomized double-blind placebo-controlled cross-over study. Healthy adults ingested preparations of living and heat-killed L. plantarum bacteria. Biopsies were taken from the intestinal duodenal mucosa and altered expression profiles were analyzed using whole-genome microarrays and by biological pathway reconstructions. Expression profiles of human mucosa displayed striking differences in modulation of NF-kappaB-dependent pathways, notably after consumption of living L. plantarum bacteria in different growth phases. Our in vivo study identified mucosal gene expression patterns and cellular pathways that correlated with the establishment of immune tolerance in healthy adults. PMID- 19190179 TI - Pituitary adenylyl cyclase-activating polypeptide is an intrinsic regulator of Treg abundance and protects against experimental autoimmune encephalomyelitis. AB - Pituitary adenylyl cyclase-activating polypeptide (PACAP) is a widely expressed neuropeptide originally discovered in the hypothalamus. It closely resembles vasoactive intestinal peptide (VIP), a neuropeptide well known to inhibit macrophage activity, promote Th2-type responses, and enhance regulatory T cell (Treg) production. Recent studies have shown that administration of PACAP, like VIP, can attenuate dramatically the clinical and pathological features of murine models of autoimmune diseases such as experimental autoimmune encephalomyelitis (EAE) and collagen-induced arthritis. However, specific roles (if any) of endogenous VIP and PACAP in the protection against autoimmune diseases have not been explored. Here, we subjected PACAP-deficient mice to myelin oligodendrocyte glycoprotein (MOG(35-55))-induced EAE. MOG immunization of PACAP-deficient mice triggered heightened clinical and pathological manifestations of EAE compared to wild-type mice. The increased sensitivity was accompanied by enhanced mRNA expression of proinflammatory cytokines (TNFalpha, IL-6, IFN-gamma, IL-12p35, IL 23p19, and IL-17), chemokines (MCP-1/CCL2, MIP-1alpha/CCL3, and RANTES/CCL5), and chemotactic factor receptors (CCR1, CCR2, and CCR5), but downregulation of the anti-inflammatory cytokines (IL-4, IL-10, and TGF-beta) in the spinal cord. Moreover, the abundance of CD4(+)CD25(+)FoxP3(+) Tregs in lymph nodes and levels of FoxP3 mRNA in the spinal cord were also diminished. The reduction in Tregs was associated with increased proliferation and decreased TGF-beta secretion in lymph node cultures stimulated with MOG. These results demonstrate that endogenous PACAP provides protection in EAE and identify PACAP as an intrinsic regulator of Treg abundance after inflammation. PMID- 19190180 TI - Tri-split tRNA is a transfer RNA made from 3 transcripts that provides insight into the evolution of fragmented tRNAs in archaea. AB - Transfer RNA (tRNA) is essential for decoding the genome sequence into proteins. In Archaea, previous studies have revealed unique multiple intron-containing tRNAs and tRNAs that are encoded on 2 separate genes, so-called split tRNAs. Here, we discovered 10 fragmented tRNA genes in the complete genome of the hyperthermoacidophilic Archaeon Caldivirga maquilingensis that are individually transcribed and further trans-spliced to generate all of the missing tRNAs encoding glycine, alanine, and glutamate. Notably, the 3 mature tRNA(Gly)'s with synonymous codons are created from 1 constitutive 3' half transcript and 4 alternatively switching transcripts, representing tRNA made from a total of 3 transcripts named a "tri-split tRNA." Expression and nucleotide sequences of 10 split tRNA genes and their joined tRNA products were experimentally verified. The intervening sequences of split tRNA have high identity to tRNA intron sequences located at the same positions in intron-containing tRNAs in related Thermoproteales species. This suggests that an evolutionary relationship between intron-containing and split tRNAs exists. Our findings demonstrate the first example of split tRNA genes in a free-living organism and a unique tri-split tRNA gene that provides further insight into the evolution of fragmented tRNAs. PMID- 19190181 TI - Following movement of the L1 stalk between three functional states in single ribosomes. AB - The L1 stalk is a mobile domain of the large ribosomal subunit E site that interacts with the elbow of deacylated tRNA during protein synthesis. Here, by using single-molecule FRET, we follow the real-time dynamics of the L1 stalk and observe its movement relative to the body of the large subunit between at least 3 distinct conformational states: open, half-closed, and fully closed. Pretranslocation ribosomes undergo spontaneous fluctuations between the open and fully closed states. In contrast, posttranslocation ribosomes containing peptidyl tRNA and deacylated tRNA in the classical P/P and E/E states, respectively, are fixed in the half-closed conformation. In ribosomes with a vacant E site, the L1 stalk is observed either in the fully closed or fully open conformation. Several lines of evidence show that the L1 stalk can move independently of intersubunit rotation. Our findings support a model in which the mobility of the L1 stalk facilitates binding, movement, and release of deacylated tRNA by remodeling the structure of the 50S subunit E site between 3 distinct conformations, corresponding to the E/E vacant, P/E hybrid, and classical states. PMID- 19190182 TI - Systems-level analysis of cell-specific AQP2 gene expression in renal collecting duct. AB - We used a systems biology-based approach to investigate the basis of cell specific expression of the water channel aquaporin-2 (AQP2) in the renal collecting duct. Computational analysis of the 5'-flanking region of the AQP2 gene (Genomatix) revealed 2 conserved clusters of putative transcriptional regulator (TR) binding elements (BEs) centered at -513 bp (corresponding to the SF1, NFAT, and FKHD TR families) and -224 bp (corresponding to the AP2, SRF, CREB, GATA, and HOX TR families). Three other conserved motifs corresponded to the ETS, EBOX, and RXR TR families. To identify TRs that potentially bind to these BEs, we carried out mRNA profiling (Affymetrix) in mouse mpkCCDc14 collecting duct cells, revealing expression of 25 TRs that are also expressed in native inner medullary collecting duct. One showed a significant positive correlation with AQP2 mRNA abundance among mpkCCD subclones (Ets1), and 2 showed a significant negative correlation (Elf1 and an orphan nuclear receptor Nr1h2). Transcriptomic profiling in native proximal tubules (PT), medullary thick ascending limbs (MTAL), and IMCDs from kidney identified 14 TRs (including Ets1 and HoxD3) expressed in the IMCD but not PT or MTAL (candidate AQP2 enhancer roles), and 5 TRs (including HoxA5, HoxA9 and HoxA10) expressed in PT and MTAL but not in IMCD (candidate AQP2 repressor roles). In luciferase reporter assays, overexpression of 3 ETS family TRs transactivated the mouse proximal AQP2 promoter. The results implicate ETS family TRs in cell-specific expression of AQP2 and point to HOX, RXR, CREB and GATA family TRs as playing likely additional roles. PMID- 19190183 TI - Inherent flexibility determines the transition mechanisms of the EF-hands of calmodulin. AB - We explore how inherent flexibility of a protein molecule influences the mechanism controlling allosteric transitions by using a variational model inspired from work in protein folding. The striking differences in the predicted transition mechanism for the opening of the two domains of calmodulin (CaM) emphasize that inherent flexibility is key to understanding the complex conformational changes that occur in proteins. In particular, the C-terminal domain of CaM (cCaM), which is inherently less flexible than its N-terminal domain (nCaM), reveals "cracking" or local partial unfolding during the open/closed transition. This result is in harmony with the picture that cracking relieves local stresses caused by conformational deformations of a sufficiently rigid protein. We also compare the conformational transition in a recently studied even-odd paired fragment of CaM. Our results rationalize the different relative binding affinities of the EF-hands in the engineered fragment compared with the intact odd-even paired EF-hands (nCaM and cCaM) in terms of changes in flexibility along the transition route. Aside from elucidating general theoretical ideas about the cracking mechanism, these studies also emphasize how the remarkable intrinsic plasticity of CaM underlies conformational dynamics essential for its diverse functions. PMID- 19190184 TI - Transient ciliogenesis involving Bardet-Biedl syndrome proteins is a fundamental characteristic of adipogenic differentiation. AB - Bardet-Biedl syndrome (BBS) is an inherited ciliopathy generally associated with severe obesity, but the underlying mechanism remains hypothetical and is generally proposed to be of neuroendocrine origin. In this study, we show that while the proliferating preadipocytes or mature adipocytes are nonciliated in culture, a typical primary cilium is present in differentiating preadipocytes. This transient cilium carries receptors for Wnt and Hedgehog pathways, linking this organelle to previously described regulatory pathways of adipogenesis. We also show that the BBS10 and BBS12 proteins are located within the basal body of this primary cilium and inhibition of their expression impairs ciliogenesis, activates the glycogen synthase kinase 3 pathway, and induces peroxisome proliferator-activated receptor nuclear accumulation, hence favoring adipogenesis. Moreover, adipocytes derived from BBS-patients' dermal fibroblasts in culture exhibit higher propensity for fat accumulation when compared to controls. This strongly suggests that a peripheral primary dysfunction of adipogenesis participates to the pathogenesis of obesity in BBS. PMID- 19190185 TI - Wildfire responses to abrupt climate change in North America. AB - It is widely accepted, based on data from the last few decades and on model simulations, that anthropogenic climate change will cause increased fire activity. However, less attention has been paid to the relationship between abrupt climate changes and heightened fire activity in the paleorecord. We use 35 charcoal and pollen records to assess how fire regimes in North America changed during the last glacial-interglacial transition (15 to 10 ka), a time of large and rapid climate changes. We also test the hypothesis that a comet impact initiated continental-scale wildfires at 12.9 ka; the data do not support this idea, nor are continent-wide fires indicated at any time during deglaciation. There are, however, clear links between large climate changes and fire activity. Biomass burning gradually increased from the glacial period to the beginning of the Younger Dryas. Although there are changes in biomass burning during the Younger Dryas, there is no systematic trend. There is a further increase in biomass burning after the Younger Dryas. Intervals of rapid climate change at 13.9, 13.2, and 11.7 ka are marked by large increases in fire activity. The timing of changes in fire is not coincident with changes in human population density or the timing of the extinction of the megafauna. Although these factors could have contributed to fire-regime changes at individual sites or at specific times, the charcoal data indicate an important role for climate, and particularly rapid climate change, in determining broad-scale levels of fire activity. PMID- 19190186 TI - Determining the scale of the Bicoid morphogen gradient. AB - Bicoid is a morphogen that sets up the anterior-posterior axis in early Drosophila embryos. Although the form of the Bicoid profile is consistent with a simple diffusion/degradation model, the observed length scale is much larger than should be expected based on the measured diffusion rate. Here, we study two possible mechanisms that could, in principle, affect this gradient and, hence, address this disagreement. First, we show that including trapping and release of Bicoid by the nuclei during cleavage cycles does not alter the morphogen length scale. More crucially, the inclusion of advective transport due to cytoplasmic streaming can have a large effect. Specifically, we build a simple model based on the (limited) experimental data and show that such a flow can lead to a Bicoid profile that is consistent with various experimental features. Specifically, the observed length scale is obtained, a steady profile is established, and improved scaling between embryos of different lengths is demonstrated. PMID- 19190187 TI - Prediction of membrane protein structures with complex topologies using limited constraints. AB - Reliable structure-prediction methods for membrane proteins are important because the experimental determination of high-resolution membrane protein structures remains very difficult, especially for eukaryotic proteins. However, membrane proteins are typically longer than 200 aa and represent a formidable challenge for structure prediction. We have developed a method for predicting the structures of large membrane proteins by constraining helix-helix packing arrangements at particular positions predicted from sequence or identified by experiments. We tested the method on 12 membrane proteins of diverse topologies and functions with lengths ranging between 190 and 300 residues. Enforcing a single constraint during the folding simulations enriched the population of near native models for 9 proteins. In 4 of the cases in which the constraint was predicted from the sequence, 1 of the 5 lowest energy models was superimposable within 4 A on the native structure. Near-native structures could also be selected for heme-binding and pore-forming domains from simulations in which pairs of conserved histidine-chelating hemes and one experimentally determined salt bridge were constrained, respectively. These results suggest that models within 4 A of the native structure can be achieved for complex membrane proteins if even limited information on residue-residue interactions can be obtained from protein structure databases or experiments. PMID- 19190188 TI - Variations on a theme. PMID- 19190189 TI - A little-known aspect of Arthur Conan Doyle (1859-1930): the call of India and a debt to Walter Scott (1771-1832). AB - This paper recalls the early life of Dr Arthur Conan Doyle when his writing centred briefly on India. The significance of a young female skeleton given to the museum of the Royal College of Surgeons of Edinburgh in 1879 is reviewed. Morphometric and genetic evidence is provided to show that the skeleton originated in the Andaman Islands. It is suggested that Doyle saw it during his undergraduate or early postgraduate years, leading him to introduce an Andaman Islander into his novel The Sign of the Four, published in 1890. Like his inspiring predecessor Walter Scott, Doyle wrote of India but did not visit the country: both authors learned indirectly of the Indian Raj and the Indian Medical Service. Doyle knew of the convict colony established after the Sepoy Mutiny of 1857 at Port Blair, capital of the Andamans, but the reason he chose an Islander to commit murder in London has, until now, remained contentious. PMID- 19190190 TI - Leonardo Da Vinci (1452-1519) as a stroke victim: hemiparesis: a result of a vegetarian diet? PMID- 19190191 TI - Professor Arthur Norman Exton-Smith CBE MA MD FRCP (1920-90): distinguished geriatrician and gerontologist. AB - Professor Norman Exton-Smith was a highly respected, distinguished postwar consultant geriatrician with a worldwide reputation. He devoted his life to improving the medical care of elderly people and researching age-related decline in physical function, particularly thermoregulation and postural balance. He established thriving clinical and research departments at St Pancras Hospital, London. Many of his junior medical staff became well-known geriatricians. He published and lectured extensively, organized many meetings and conferences, and was advisor to the Department of Health and Social Security for many years. He was a valued authority on geriatric medicine within the Royal College of Physicians of London and a major influence in the British Geriatrics Society (BGS) of which he was Secretary and later the President. PMID- 19190192 TI - Moses Maimonides (1135-1204). PMID- 19190193 TI - Sir Stanford Cade KBE CB FRCS (1895-1973): a pioneer in the modern treatment of cancer. AB - Stanford Cade, born in Tsarist Russia, trained in Medicine first in Brussels and then in London at King's College and Westminster Hospital. His potential as a brilliant clinician was recognized by his appointment to the surgical staff at Westminster at the early age of 29. Here he was one of the first in the UK to use radium in the treatment of a wide variety of tumours. His interests covered the broad spectrum of malignant diseases including the head and neck, breast, bone and soft tissues. He was an early exponent of the multidisciplinary approach to the management of cancer. PMID- 19190194 TI - Dr Adam Zamenhof (1888-1940) and his insight into ophthalmology. AB - Adam Zamenhof was greatly influenced by his father, Ludwik Zamenhof, who designed the international language Esperanto. Like his father, he became an ophthalmologist and joined the Esperanto movement. He published in the field of ophthalmology and was soon chosen as head of an ophthalmology department. He subsequently became Chief of the Orthodox Jewish Hospital at Czystem in Warsaw. He was active in the leadership of the Bialystok-Warsaw Chamber of Medical Doctors. He perished in the Nazi Holocaust (Shoah) but all Zamenhof's ideals that Adam served as a doctor and social activist remain still alive. PMID- 19190195 TI - Ignac Fulop Semmelweis (1818-65). PMID- 19190196 TI - Dr William Sargant (1907-88) and the emergence of physical treatments in British psychiatry. AB - Dr William Walters Sargant stands out as a firm champion of physical treatments in 20th century British psychiatry. Some saw his ultra-physical approach as evidence of the progress that the speciality of psychiatry had made in moving on from its unscientific beginnings in the 19th century. Other psychiatrists, however, maintained that Sargant's practices were too 'surgical', immoral even. Sargant was a flag-bearer for the new treatment methods of insulin coma, electroconvulsive therapy (ECT), psychosurgery and intensive drug therapy. He rebutted the claims of clinicians who argued for a more psychologically or socially driven approach. Moreover, he advocated the eradication of memories from the mind by physical means rather than teasing them out by Freudian analysis. PMID- 19190197 TI - Walter Russell Brain (1895-1966), Baron Brain of Eynsham, Lord Brain MA DM DSc LLD DCL FRCP FRCS FRCOG FRCPEd. FRCP.Glas FRACP FACP FCPSA FRS. PMID- 19190198 TI - Intensive care 1650: the revival of Anne Greene (c. 1628-59). AB - On a cold December day in 1650, 22-year-old Anne Greene was hanged in Oxford. When taken down after half an hour, she was found to show signs of life and over the next few days William Petty (1623-87), Thomas Willis (1621-75), Ralph Bathurst (1620-74) and Henry Clerke (1622-87) ministered to her full recovery. She was later pardoned of the charge of infanticide and, with the coffin wherein she had lain as a trophy, went into the country, became the subject not only of a prose and poetic narrative but also of a woodcut. Anne married happily, bore three children and lived until 1659. A combination of low-body temperature and external (pedal) cardiac massage after her failed execution, it is suggested, helped to keep her alive until the arrival of the physicians who had come to make an anatomical dissection but serendipitously won golden opinions. PMID- 19190199 TI - John Hyacinth de Magellan (1722-90): 18th century physicist with views on medical matters. AB - John Hyacinth de Magellan, whose Portuguese name was Joao Hyacintho de Magalhaens, though not a doctor nevertheless had many contacts with doctors and showed a genuine interest in disseminating medical news to his many friends and correspondents in Europe. The abundant and less formal correspondence with his friend Ribeiro Sanches forms the greater part of the work but in letters to other correspondents, including Trudaine de Montigny, Condorcet, Volta, J A Euler, Fabroni and Johann III Bernoulli, we find comments on medical subjects. The Sanches letters are particularly interesting because they are private, friend-to friend letters that convey spontaneous and sincere thoughts and feelings. PMID- 19190200 TI - Nikolai Konstantinovich Kulchitsky (1856-1925). AB - Nikolai Kulchitsky is best remembered for his identification of the Kulchitsky (enterochromaffin) cell. His life spanned a teaching and scientific career at Kharkov University, employment as the Imperial Minister of Education for all Russia, work in a soap factory and flight from the Russian Revolution to London, and a position at the University College with Elliot Smith. His subsequent contributions to the anatomic delineation of dual nerve-endings in the muscle were highly regarded, although his identification of the enterochromaffin cell (1897) remains his enduring scientific legacy. The observation of a cardinal neuroendocrine cell of the gut formed the basis for the subsequent delineation of the diffuse neuroendocrine system and provided the cellular framework on which the discipline of gut neuroendocrinology would be established. Kulchitsky's mysterious demise in a bizarre lift-shaft accident at UCL on his 69th birthday tragically terminated a life of service to science. PMID- 19190201 TI - Conn's syndrome. PMID- 19190202 TI - Gertrude Stein (1874-1946) and the nucleus of Darkschewitsch: a discursive commentary. AB - Gertrude Stein is an icon of American literature whose scientific and medical background has become shrouded in obscurity. As an undergraduate at Radcliffe she was strongly influenced by William James and published two papers on motor automatism in the Psychological Review. As a medical student at Johns Hopkins University, her research on the nucleus of Darkschewitsch was quoted in Lewellys F Barker's acclaimed textbook on neuroanatomy; Stein's first book appearance. The background of the Russian neurologist Liverji O Darkschewitsch, little known in the West, is explored particularly in regard to his relationship and collaboration with Sigmund Freud whose letters provide considerable insight. Gertrude Stein failed to graduate with her class of 1901 at Johns Hopkins and soon after departed for an expatriate life in Europe devoted to art and literature. PMID- 19190203 TI - Letter to the editor. PMID- 19190204 TI - Asymptomatic, transient complete heart block in a pediatric patient with Lyme disease. AB - Lyme Disease, caused by the spirochete Borrellia burgdorferi, is the most common vector-borne disease in the United States. Clinically, it primarily affects the skin, joints, nervous system, and heart. Lyme carditis occurs in 4%-10% of adults with Lyme disease. Transient variable-level atrioventricular blocks, occurring in 77% of adults with Lyme carditis, are the most common cardiac manifestation. Up to 50% of Lyme carditis patients may develop complete heart block. The incidence of Lyme carditis in the pediatric population is not well established. We present a pediatric patient with a transient asymptomatic complete heart block resulting from Lyme carditis, an under-recognized complication of Lyme disease in the pediatric population. PMID- 19190205 TI - Malarial retinopathy in cerebral malaria. PMID- 19190206 TI - Efficacy of cryotherapy versus intralesional meglumine antimoniate (glucantime) for treatment of cutaneous leishmaniasis in children. AB - We compared intralesional glucantine and cryotherapy for treatment of children with cutaneous leishmaniasis in Iran. We observed that cryotherapy is an effective treatment for cutaneous leishmaniasis in children. No serious post treatment side effects were observed in either group. At six months of follow-up, no recurrence of disease was observed in cured patients in either group. Because of its simplicity, lower cost, low rate of serious complications, and greater tolerability, cryotherapy should be recommended as an appropriate alternative treatment for leishmaniasis in children. PMID- 19190207 TI - Case report: nitazoxanide for treatment of refractory bony hydatid disease. AB - Although nitazoxanide has been shown to have activity against Echinococcus multilocularis in animal studies and against E. granulosus in vitro, its use in treatment of human cystic echinococcosis has not been reported. We report a case of progressive bony hydatid disease treated with nitazoxanide that showed a clinical and radiologic response. The patient had a 40-year history of hydatid disease involving the left hip. Despite prolonged courses of albendazole and praziquantel, she had progressive disease with extensive involvement of the left hemipelvis and adjacent soft tissue cysts. She was treated with nitazoxanide, 500 mg twice a day for 3 months in combination with albendazole. The clinical response was supported by imaging showing marked improvement in the soft tissue cysts, with stable disease in the bony pelvis. Although further studies are required, this case suggests that nitazoxanide may be an effective treatment option in hydatid disease, particularly in patients with progressive disease who are receiving conventional therapy. PMID- 19190208 TI - Case report: rectal perforation caused by Schistosoma haematobium. AB - A 31-year-old woman from Cameroon was admitted to the University of Strasbourg Hospital in December 2007 with pelvic pain and fever that developed over three days. Her condition rapidly worsened and she underwent emergency exploratory celioscopy. Surgeons found peritoneal and retrouterine abscesses. The high rectum had a 4-cm perforation with infiltrated, friable, and irregular edges. A biopsy specimen of this pseudotumoral specimen showed many Schistosoma haematobium eggs with an inflammatory reaction surrounding the eggs. The patient was treated with praziquantel (40 mg/kg/day) for 5 days and a 4-week course of antibiotic therapy. Her progress was good and digestive continuity surgery was performed four months later. Schistosomiasis frequently involves rectal mucosa, but perforation is unusual. Our review of the literature found only two cases of colon perforation associated with S. mansoni infection. To our knowledge, this is the first case of rectal perforation caused by S. haematobium described in the literature. PMID- 19190209 TI - Melioidosis after brief exposure: a serologic survey in US Marines. AB - Melioidosis is endemic to Southeast Asia. The incidence of infection in visitors is not well known, especially for short visits. Thirteen (38%) of 34 previously unexposed US Marines had positive serology after 2 weeks in Thailand, and one developed acute disseminated disease. Asymptomatic infection with Burkholderia pseudomallei may be common, even from brief exposures. PMID- 19190210 TI - Short report: case of gnathostomiasis in Beijing, China. AB - A 59-year-old woman sought treatment of for creeping eruption and erythematous plaques associated with high fever and systemic symptoms. She had a history of eating undercooked freshwater fish raised in Beijing 10 days before admission and reported no travel during the previous year. Blood examination showed eosinophila and ultrasonography detected multiple hypoechoic areas in the liver and spleen. Western blot test detected specific antibodies to the larvae of Gnathostoma spinigerum. Cutaneous and visceral larva migrations associated with G. spinigerum were diagnosed. Twenty-three cases in 12 provinces have been reported in the Chinese literature but none have been reported in English. PMID- 19190211 TI - Methemoglobinemia and adverse events in Plasmodium vivax malaria patients associated with high doses of primaquine treatment. AB - Primaquine (PQ) is recommended to prevent relapses in patients with Plasmodium vivax malaria infection. However, treatment with PQ causes methemoglobinemia. In this study, we measured the methemoglobin (MetHB) levels in three groups of subjects who received PQ treatment at 0.58, 0.83, or 1.17 mg/kg/d. A total of 112 subjects were studied. MetHB levels were detected at > or = 4% in 46-50% 1 day after PQ treatment in all three groups and 4-9% of subjects had MetHB levels > or = 4% 15 days after treatment. Only subjects receiving the highest doses of PQ had mild and brief adverse events, and 17% of them were associated with treatment. We conclude that when PQ is administered under certain conditions (i.e., normal glucose-6-phosphate dehydrogenase activity, in non-pregnant subjects and with a light meal), daily doses as high as 1.17 mg/kg do not represent a serious risk of high MetHB levels to patients. PMID- 19190212 TI - Severe Plasmodium vivax malaria: a report on serial cases from Bikaner in northwestern India. AB - Epidemiologic studies and clinical description of severe Plasmodium vivax malaria in adults living in malaria-endemic areas are rare and more attention is needed to understand the dynamics and its interaction with the immune system. This observational study included 1,091 adult patients admitted to medical wards of S. P. Medical College and associated group of hospitals in Bikaner, India from September 2003 through December 2005. The diagnosis of P. vivax malaria was established by peripheral blood film (PBF), rapid diagnostic test (RDT), and polymerase chain reaction (PCR), and severe malaria was categorized as per World Health Organization guidelines. Of 1,091 patients with malaria, 635 had P. falciparum malaria and 456 had P. vivax malaria. Among patients with severe manifestations, 40 had evidence of monoinfection of P. vivax malaria diagnosed by PBF, RDT, and PCR. Complications observed were hepatic dysfunction and jaundice in 23 (57.5%) patients, renal failure in 18 (45%) patients, severe anemia in 13 (32.5%) patients, cerebral malaria in 5 patients (12.5%), acute respiratory distress syndrome in 4 patients (10%), shock in 3 patients (7.5%), and hypoglycemia in 1 (2.5%) patient. Thrombocytopenia was observed in 5 (12.5%) patients, and multi-organ dysfunction was detected in 19 (47.5%) patients. Further large-scale multicentric epidemiologic studies are needed to define the basic pathology of this less known entity. PMID- 19190213 TI - Short report: comparison of chlorproguanil-dapsone with a combination of sulfadoxine-pyrimethamine and chloroquine in children with malaria in northcentral Nigeria. AB - Effective and affordable treatment of malaria is critical in the face of resistance of Plasmodium falciparum to chloroquine (CQ) and sulfadoxine pyrimethamine (SP). We conducted a randomized controlled trial comparing the efficacy of chlorproguanil-dapsone (CD) with a combination SP plus CQ in children in Nigeria less than five years of age with malaria. Of 264 children enrolled, 122 (89.7%) and 118 (92.2%) completed the study in the SP + CQ and CD groups, respectively. By day 3, 96 (78.7%) and 94 (79.7%) had cleared their parasitemia (P = 0.79), and 107 (87.7%) and 109 (92.4%) were symptom free (P = 0.32) in the SP + CQ and CD groups, respectively. Adequate clinical and parasitologic response at day 14 occurred in 111 (94.1%; 95% confidence interval [CI] = 91.6-95.7%) in the CD group and 113 (92.6%; 95% CI = 89.9-94.3%) in the SP + CQ group (P = 0.85). SP + CQ and CD had similar antimalarial efficacy and still provide affordable treatment of uncomplicated malaria in northcentral Nigeria. PMID- 19190214 TI - Influence of wasting and stunting at the onset of the rainy season on subsequent malaria morbidity among rural preschool children in Senegal. AB - In sub-Saharan Africa, malaria and malnutrition are major causes of morbidity and mortality in children less than five years of age. To explore the impact of malnutrition on subsequent susceptibility to malaria, a cohort of 874 rural preschool children in Senegal was followed-up during one malaria transmission season from July through December. Data on nutritional status and Plasmodium falciparum parasitemia were collected at baseline. Malaria morbidity was monitored through weekly home visits. Wasted children (weight-for-height z-score < -2) were at lower risk of having at least one subsequent clinical malaria attack (odds ratio = 0.33; 95% confidence interval = 0.13-0.81, P = 0.02), whereas stunting (height-for-age z-score < -2) or being underweight (weight-for age z-score < -2) was not associated with clinical malaria. Although non biological explanations such as overprotection of wasted children by their mothers should be considered, immunomodulation according to nutritional status could explain the lower risk of malaria attack among wasted children. PMID- 19190215 TI - Assessment of insecticide-treated bednet use among children and pregnant women across 15 countries using standardized national surveys. AB - Impact of insecticide-treated bednets (ITNs) on preventing malaria may be minimized if they are not used by vulnerable populations. Among ITN-owning households from 15 standardized national surveys from 2003 to 2006, we identify factors associated with ITN use among children younger than 5 years of age and make comparisons of ITN use among children and pregnant women across countries. Within ITN-owning households, many children and pregnant women are still not using them. Between-country analysis with linear regression showed child ITN use increases as intra-household access to ITNs increases (P = 0.020, R2 = 0.404), after controlling for season and survey year. Results from within-country logistic regression analyses were consistent with between-country analysis showing intra-household access to ITNs is the strongest and most consistent determinant of use among children. The gaps in ITN use and possession will likely persist in the absence of achieving a ratio of no more than two people per ITN. PMID- 19190216 TI - Short report: chloroquine-resistant Plasmodium vivax in the Republic of Korea. AB - The number of Plasmodium vivax malaria patients in the Republic of Korea and North Korea since the re-emergence of malaria in 1993 is estimated to be approximately one million. To cope with this situation, the Army of the Republic of Korea has performed chemoprophylaxis with hydroxychloroquine and primaquine since 1997. The cumulative number of soldiers in the Army of the Republic of Korea given chemoprophylaxis exceeded 1.4 million by 2007. Extensive chemoprophylaxis contributed to preventing a rapid increase of malaria patients in the Army of the Republic of Korea, but increased the possibility of the occurrence of chloroquine (CQ)-resistant P. vivax strains. In this study, treatment responses of P. vivax malaria patients in the Republic of Korea monitored during 2003-2007, and CQ resistance was confirmed in 2 of 484 enrolled patients. Our results are the first report of CQ-resistant P. vivax in a temperate region of Asia. Continuous surveillance is warranted to monitor the change in CQ resistance frequency of P. vivax in the Republic of Korea. PMID- 19190218 TI - Studies on the Salvador I strain of Plasmodium vivax in non-human primates and anopheline mosquitoes. AB - A review is presented on studies conducted in New World monkeys and chimpanzees with the Salvador I strain of Plasmodium vivax. This isolate has been adapted to Aotus and Saimiri (squirrel) monkeys and developed as a model for the testing of antimalarial vaccines. After the injection of 10,000 sporozoites, the median prepatent period in S. boliviensis monkeys was 21.5 days. In 103 sporozoite induced infections in splenectomized monkeys, the median maximum parasite count ranged from 2,139 to 202,368/microL, with a median maximum parasite count of 48,174/microL. Median maximum parasite counts in Aotus lemurinus griseimembra, A. nancymaae, A. azarae boliviensis, and A. vociferans monkeys were 19,902, 18,390, 21,420, and 18,210/microL, respectively and ranged from 124 to 156,000/microL. Mosquito infections were readily obtained in different species of Anopheles mosquitoes. The S. boliviensis monkey and Salvador I strain seems suitable for the testing of sporozoite and liver stage vaccines but not for blood-stage vaccines against P. vivax unless adapted further in spleen-intact Saimiri boliviensis monkeys. PMID- 19190217 TI - Genetic variation among Plasmodium vivax isolates adapted to non-human primates and the implication for vaccine development. AB - Plasmodium vivax Duffy binding protein (DBP) is vital for parasite development, thereby making this molecule a good vaccine candidate. Preclinical development of a P. vivax vaccine often involves use of primate models prior to testing efficacy in humans, but primate isolates are poorly characterized. We analyzed the complete gene coding for the DBP in several P. vivax isolates that are used for experimental primate infections and compared these sequences with the Salvador I DBP isolate, which is being used for vaccine development. Our results affirm that primate-adapted isolates are genetically similar to P. vivax circulating in humans, but variability is greatest in the putative target of protective antibodies. In addition, some P. vivax isolates contain multiple genetically different clones. Testing a DBP vaccine may therefore be complicated by heterogeneity and diversity of the P. vivax isolates available for in vivo challenge. PMID- 19190219 TI - Short report: a multiplex PCR assay for simultaneous genotyping of kdr and ace-1 loci in Anopheles gambiae. AB - The selection of insecticide-resistant genotypes in Anopheles gambiae, the most important malaria vector in Africa, makes disease control problematic in several endemic areas. The early detection and monitoring of resistance associated mutations in field mosquito populations is essential for the application of successful insecticide-based control interventions. Currently, the surveillance of these mutations is performed using individual assays, some of which require sophisticated and expensive equipment. Here we describe a novel multiplex polymerase chain reaction-based assay for detecting simultaneously the five single nucleotide polymorphisms in the voltage-gated sodium channel and the ace-1 genes, which have been associated with the mosquito response to most commonly used insecticides. PMID- 19190220 TI - Short report: concurrent detection of Trypanosoma cruzi lineages I and II in domestic Triatoma dimidiata from Guatemala. AB - The agent of Chagas disease, Trypanosoma cruzi, is phylogenetically divided into two lineages, T. cruzi I (TCI) and II (TCII). TCI is found in sylvatic and domestic habitats across South America. Despite a high prevalence of TCII in domestic habitats in South America, it has been rarely found in domestic habitats in Central America and Mexico. This may be caused by limitations in detection tests previously used. A modified hemi-nested polymerase chain reaction assay was developed to improve detection of TCI and TCII mini-exon genes. This method detected TCI and II concurrently in 84% of 44 cultured isolates from Triatoma dimidiata specimens collected inside homes across the disease-endemic area in Guatemala. PMID- 19190221 TI - Short report: serologic evidence of human ehrlichiosis in Peru. AB - A serosurvey for human ehrlichiosis caused by Ehrlichia chaffeensis and Anaplasma phagocytophilum was performed in different regions of Peru by using indirect immunofluorescence assays (IFAs). Regions included an urban community in a shantytown in Lima (Pampas) and three rural communities located on the northern coast of Peru (Cura Mori), in the southern Peruvian Andes (Cochapata), and in the Peruvian jungle region (Santo Tomas). An overall E. chaffeensis seroprevalence of 13% (21 of 160) was found by IFA. Seroprevalences in females and males was 15% (16 of 106) and 9% (5 of 53), respectively. Seroprevalences in Cura Mori, Cochapata, Pampas, and Santo Tomas were 25% (10 of 40), 23% (9 of 40), 3% (1 of 40), and 3% (1 of 40), respectively. Seroprevalences in Cura Mori and Cochapata were significantly higher than in Santo Tomas or Pampas (P < 0.01). No sera were reactive to A. phagocytophilum. These findings suggest that human infection with E. chaffeensis occurs in Peru. Further studies are needed to characterize Ehrlichia species in Peru, their vectors and their clinical significance. PMID- 19190222 TI - Genetic diversity of Trichomonas vaginalis clinical isolates determined by EcoRI restriction fragment length polymorphism of heat-shock protein 70 genes. AB - Restriction fragment length polymorphism (RFLP) analysis using a multilocus heat inducible cytoplasmic heat-shock protein 70 (Hsp70) hybridization probe with EcoRI-digested genomic DNA was used in molecular typing of 129 Trichomonas vaginalis isolates. Results indicate that Trichomonas organisms exhibit considerable polymorphism in their Hsp70 RFLP patterns. Analysis of seven American Type Culture Collection reference strains and 122 clinical isolates, including 84 isolates from Jackson, Mississippi, 18 isolates from Atlanta, Georgia, and 20 isolates from throughout the United States, showed 105 distinct Hsp70 RFLP pattern subtypes for Trichomonas. Phylogenetic analysis of the Hsp70 RFLP data showed that the T. vaginalis isolates were organized into two clonal lineages. These results illustrate the substantial genomic diversity present in T. vaginalis and indicate that a large number of genetically distinct Trichomonas isolates may be responsible for human trichomoniasis in the United States. PMID- 19190223 TI - Arthritogenicity of Borrelia burgdorferi and Borrelia garinii: comparison of infection in mice. AB - Arthritogenicity, as determined by joint swelling and synovial histology, was compared between or within two Borrelia genospecies that cause Lyme arthritis in humans. The spirochete burden in bladder tissue (a site of spirochete persistence) was documented by quantitative polymerase chain reaction, and immune responses were analyzed. In C3H/HeJ mice, three B. burgdorferi isolates and two of the three B. garinii isolates induced severe arthritis and swelling. Previous designation as invasive or noninvasive B. garinii, or RNA spacer type of B. burgdorferi did not determine arthritis severity induced by isolates. Compared with the other five isolates, the B. garinii PBi isolate induced significantly less arthritis, a lower humoral immune response, and persisted at a much lower level in bladder tissue. However, B. garinii PBi isolates induced similar Borrelia antigen-specific inflammatory T cell responses from the local draining lymph node. Thus, diverse B. burgdorferi and B. garinii isolates were highly arthritogenic in immune competent mice. PMID- 19190224 TI - Short report: time course of hematogenous dissemination of Francisella tularensis A1, A2, and Type B in laboratory mice. AB - Tularemia is a tick-borne zoonotic bacterial disease. In the United States, human tularemia infections are caused by Francisella tularensis subspecies tularensis (Type A, clades A1 and A2) or F. tularensis subspecies holarctica (Type B). We developed a mouse model that can be used to study the ability of ticks to acquire and transmit fully virulent strains of F. tularensis (A1, A2, and Type B). We showed that 1) bacteremia was evident by 2 days post-infection (dpi) for A1, A2, and B, 2) bacteremia was expected to reach levels of > 10(8) cfu/mL by 3 dpi for A1 and A2 but not until 4 dpi for Type B, and 3) illness onset was delayed for mice exposed to Type B compared with A1 and A2. To maximize the likelihood of ticks acquiring infection from laboratory-infected mice before they become moribund and must be euthanized, ticks should be placed on mice so that periods of rapid engorgement occur 3-4 dpi for A1 and A2 and 4-5 dpi for Type B. Rigorous experimental studies of tick vector competence and efficiency conducted under standardized conditions are required to address several significant public health issues related to preventing and controlling tularemia. Our study provides the basis for a mouse model needed as the starting point to address these questions. PMID- 19190225 TI - Short report: prevalence of Neospora caninum and Toxoplasma gondii antibodies in northern Egypt. AB - In view of the worldwide importance of Toxoplasma gondii and Neospora caninum and the limited data on the seroprevalence of these parasites in Egypt, this study aimed to estimate the prevalence of anti-T. gondii and anti-N. caninum antibodies in rabbits, cattle, and humans. We used ELISA methods based on surface antigen 2 of T. gondii (TgSAG2t) and surface antigen 1 of N. caninum (NcSAG1t). High seroprevalence of T. gondii (51.49%) was detected in pregnant women, and antibodies to N. caninum were also detected in human samples (7.92%). Anti-T. gondii or N. caninum antibodies were detected in cattle (TgSAG2t: 10.75%; NcSAG1t: 20.43%). In rabbits, only one sample was N. caninum positive (1.85%). The high prevalence of toxoplasmosis and neosporosis in cattle affects the development of the livestock industry and is also an important infective source for human infection in Egypt. PMID- 19190226 TI - Host selection by Culex pipiens mosquitoes and West Nile virus amplification. AB - Recent field studies have suggested that the dynamics of West Nile virus (WNV) transmission are influenced strongly by a few key super spreader bird species that function both as primary blood hosts of the vector mosquitoes (in particular Culex pipiens) and as reservoir-competent virus hosts. It has been hypothesized that human cases result from a shift in mosquito feeding from these key bird species to humans after abundance of the key birds species decreases. To test this paradigm, we performed a mosquito blood meal analysis integrating host feeding patterns of Cx. pipiens, the principal vector of WNV in the eastern United States north of the latitude 36 degrees N and other mosquito species with robust measures of host availability, to determine host selection in a WNV endemic area of suburban Chicago, Illinois, during 2005-2007. Results showed that Cx. pipiens fed predominantly (83%) on birds with a high diversity of species used as hosts (25 species). American robins (Turdus migratorius) were marginally overused and several species were underused on the basis of relative abundance measures, including the common grackle (Quiscalus quiscula), house sparrow (Passer domesticus), and European starling (Sturnus vulgaris). Culex pipiens also fed substantially on mammals (19%; 7 species with humans representing 16%). West Nile virus transmission intensified in July of both years at times when American robins were heavily fed upon, and then decreased when robin abundance decreased, after which other birds species were selected as hosts. There was no shift in feeding from birds to mammals coincident with emergence of human cases. Rather, bird feeding predominated when the onset of the human cases occurred. Measures of host abundance and competence and Cx. pipiens feeding preference were combined to estimate the amplification fractions of the different bird species. Predictions were that approximately 66% of WNV-infectious Cx. pipiens became infected from feeding on just a few species of birds, including American robins (35%), blue jays (17%, Cyanocitta cristata), and house finches (15%, Carpodacus mexicanus). PMID- 19190228 TI - A randomized controlled trial of the concrete biosand filter and its impact on diarrheal disease in Bonao, Dominican Republic. AB - A number of household water treatment and safe storage technologies, such as chlorine disinfection, solar disinfection, and ceramic filtration, have been documented for their ability to reduce diarrheal disease and improve microbial water quality. The biosand filter (BSF) is a promising household water treatment technology in use by > 500,000 people globally. The purpose of this research was to document the ability of BSFs to improve water quality and to reduce diarrheal disease in user compared with non-user households in a randomized controlled trial in Bonao, Dominican Republic, during 2005-2006. During the 6-month intervention period, 75 BSF households had significantly improved drinking water quality on average compared with 79 control households (P < 0.001). Based on random intercepts logistic regression, BSF households had 0.53 times the odds of diarrheal disease as control households, indicating a significant protective effect of the BSF against waterborne diarrheal disease. PMID- 19190227 TI - Potential renal sequelae in survivors of hantavirus cardiopulmonary syndrome. AB - Although other hantaviruses are associated with renal manifestations, hantavirus cardiopulmonary syndrome (HCPS) has not been associated with such sequelae. The HCPS survivors were prospectively evaluated for renal complications. Subjects underwent yearly evaluation, laboratory studies, and 24-hour urine collection. Thirty subjects were evaluated after recovery from HCPS with the first follow-up at a median of 7.4 months after discharge. Subjects were a wide age range (18-51) but had an equal gender composition. Eighteen of 30 (60%) returned for > 1 evaluation. Half (15/30) had a 24-hour urine collection with > 150 mg of total protein and 6 had > 300 mg. Seven had a Cockcroft-Gault creatinine clearance (CrClCG) < 90 mL/min/1.73 m2 and 2 were < 60. Fifty-three percent met the definition of chronic kidney disease. Those treated with extracorporeal membrane oxygenation had less renal sequelae (P = 0.035). Our data suggest that renal sequelae may occur in HCPS. Further study of renal complications of New World hantavirus infections are needed. PMID- 19190229 TI - Short report: high prevalence of serine protease autotransporter cytotoxins among strains of enteroaggregative Escherichia coli. AB - Enteroaggregative Escherichia coli (EAEC) pathogenesis is thought to comprise intestinal colonization followed by the release of enterotoxins and cytotoxins. Here, we use a polymerase chain reaction (PCR) to determine the prevalence of 10 genes encoding serine protease autotransporter toxins (SPATEs) in a collection of clinical EAEC isolates. Eighty-six percent of EAEC strains harbored genes encoding one or more class I cytotoxic SPATE proteins (Pet, Sat, EspP, or SigA). Two Class II, non-cytotoxic, SPATE genes were found among EAEC strains: pic and sepA, each originally described in Shigella flexneri 2a. Using a multiplex PCR for five SPATE genes (pet, sat, sigA, pic, and sepA), we found that most of the Shigella isolates also harbored more than one SPATE, whereas members of most other E. coli pathotypes rarely harbored a cytotoxic SPATE gene. SPATEs may be relevant to the pathogenesis of both EAEC and Shigella spp. PMID- 19190230 TI - Evaluation of interferences between dengue vaccine serotypes in a monkey model. AB - Interferences between different antigens in the same vaccine formulation have been reported for some vaccines (e.g., polio vaccines, live attenuated dengue vaccine candidates). We examined interferences between the four serotypes of ChimeriVax dengue vaccines (CYDs) in a monkey model when present within a tetravalent formulation in equal concentrations (TV-5555). Immunoassays of vaccinated non-human primates showed that serotype 4 (DEN-4), and to a lesser extent, DEN-1 were dominant in terms of neutralizing antibody levels. Parameters that affected the interferences were identified, including 1) the simultaneous administration of two complementary bivalent vaccines at separate anatomical sites drained by different lymph nodes; 2) the sequential administration of two complementary bivalent vaccines; 3) the establishment of heterologous flavivirus pre-immunity before subsequent tetravalent immunization; 4) the adaptation of formulations by decreasing the dose of the immunodominant serotype; and 5) the administration of a 1-year booster. The applicability of these data to human responses is discussed. PMID- 19190231 TI - Development of a framework for evaluating the sustainability of community-based dengue control projects. AB - There are currently no frameworks developed specifically for assessing community based dengue control project sustainability. We first review the literature for frameworks for assessing project sustainability and second validate the framework criteria against the oldest community-based intervention using Mesocyclops in Xuan Phong commune, Nam Dinh province, north Vietnam, the subject of an intervention in 1998-2000. The framework used 13 criteria, clustered into three categories: 1) maintenance of health benefits from the original project, 2) continued delivery of community activities, and 3) human resource development. To provide consistency between criteria and to allow comparison both over time and with non-intervention communes, a five-point scale for each criterion was used, with the overall sustainability score calculated as the mean of all criteria. The framework offers a practical tool for assessing sustainability, and is amenable to adaptation for specific interventions without compromising the framework as a whole. PMID- 19190232 TI - Evaluation of immunoprophylactic efficacy of Brugia malayi transglutaminase (BmTGA) in single and multiple antigen vaccination with BmALT-2 and BmTPX for human lymphatic filariasis. AB - An attempt was made to study the immunoprophylactic efficacy of recombinant Brugia malayi transglutaminase (BmTGA) as protein vaccine along with two other recombinant proteins, Brugia malayi abundant larval transcript-2 (BmALT-2) and Brugia malayi thioredoxin peroxidase (BmTPX), in single and multiple antigen form for human lymphatic filariasis. Parasite challenge studies in jirds exhibited protection of 30%, 69%, and 43% against BmTGA, BmALT-2, and BmTPX, respectively, in single antigen vaccination mode. The protective efficacy of BmTGA was enhanced significantly (74%) by immunizing the jirds in multiple antigen vaccination mode along with BmTPX, whereas immunizing with the combination of BmTGA and BmALT2 conferred only 47% protection. The same protection profiles were obtained by in vitro antibody-dependent cellular cytotoxicity, using live microfilariae and L3 stage larvae. The immune response was Th2 biased, irrespective of single or multiple vaccinations. The combination of BmTGA and BmTPX seems to be a promising vaccine candidate against lymphatic filariasis. PMID- 19190233 TI - Camostat attenuates airway epithelial sodium channel function in vivo through the inhibition of a channel-activating protease. AB - Inhibition of airway epithelial sodium channel (ENaC) function enhances mucociliary clearance (MCC). ENaC is positively regulated by channel-activating proteases (CAPs), and CAP inhibitors are therefore predicted to be beneficial in diseases associated with impaired MCC. The aims of the present study were to 1) identify low-molecular-weight inhibitors of airway CAPs and 2) to establish whether such CAP inhibitors would translate into a negative regulation of ENaC function in vivo, with a consequent enhancement of MCC. To this end, camostat, a trypsin-like protease inhibitor, provided a potent (IC(50) approximately 50 nM) and prolonged attenuation of ENaC function in human airway epithelial cell models that was reversible upon the addition of excess trypsin. In primary human bronchial epithelial cells, a potency order of placental bikunin > camostat > 4 guanidinobenzoic acid 4-carboxymethyl-phenyl ester > aprotinin >> soybean trypsin inhibitor = alpha1-antitrypsin, was largely consistent with that observed for inhibition of prostasin, a molecular candidate for the airway CAP. In vivo, topical airway administration of camostat induced a potent and prolonged attenuation of ENaC activity in the guinea pig trachea (ED(50) = 3 microg/kg). When administered by aerosol inhalation in conscious sheep, camostat enhanced MCC out to at least 5 h after inhaled dosing. In summary, camostat attenuates ENaC function and enhances MCC, providing an opportunity for this approach toward the negative regulation of ENaC function to be tested therapeutically. PMID- 19190234 TI - Carbon monoxide rescues mice from lethal sepsis by supporting mitochondrial energetic metabolism and activating mitochondrial biogenesis. AB - Use of metal carbonyl-based compounds capable of releasing carbon monoxide (CO) in biological systems have emerged as a potential adjunctive therapy for sepsis via their antioxidant, anti-inflammatory, and antiapoptotic effects. The role of CO in regulation of mitochondrial dysfunction and biogenesis associated with sepsis has not been investigated. In the present study, we employed a ruthenium based water-soluble CO carrier, tricarbonylchoro(glycinato)ruthenium (II) (CORM 3), one of the novel CO-releasing molecules (CO-RMs), to test whether CO can improve cardiac mitochondrial dysfunction and survival in peritonitis-induced sepsis. Peritonitis was performed in mice by cecal ligation and perforation. Tumor necrosis factor-alpha, interleukin-10, and nitrite/nitrate plasma levels were tested to evaluate the systemic inflammatory response. Functional mitochondrial studies included determination of membrane potential, respiration, and redox status. Oxidative stress was evaluated by measurements of mitochondrial hydrogen peroxide, carbonyl protein and GSH levels. Mitochondrial biogenesis was assessed by peroxisome proliferator-activated receptor gamma coactivator (PGC) 1alpha protein expression and mitochondrial DNA (mtDNA) copy number. The systemic inflammatory response elicited by peritonitis was accompanied by mitochondrial energetic metabolism deterioration and reduced PGC-1alpha protein expression. CORM-3 treatment in septic mice restored the deleterious effects of sepsis on mitochondrial membrane potential, respiratory control ratio, and energetics. It is interesting that administration of CORM-3 during sepsis elicited a mild oxidative stress response that stimulated mitochondrial biogenesis with PGC 1alpha protein expression and mtDNA copy number increases. Our results reveal that delivery of controlled amounts of CO dramatically reduced mortality in septic mice, indicating that CO-RMs could be used therapeutically to prevent organ dysfunction and death in sepsis. PMID- 19190235 TI - The activation of M1 muscarinic receptor signaling induces neuronal differentiation in pyramidal hippocampal neurons. AB - Muscarinic receptors have been proposed to play an important role during brain development by regulating cell survival, proliferation, and differentiation. This study investigated the effect of muscarinic receptor activation on prenatal rat hippocampal pyramidal neuron differentiation and the signal transduction pathways involved in this effect. The cholinergic agonist carbachol, after 24 h in vitro, increased the length of the axon, without affecting the length of minor neurites. Carbachol-induced axonal growth was also observed in pyramidal neurons from the neocortex but not in granule neurons from the cerebellum. The effect of carbachol was mediated by the M(1) subtype of muscarinic receptors. The Ca(2+) chelator 1,2 bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester, the two protein kinase C (PKC) inhibitors 3-[1-[3-(dimethylaminopropyl]-1H-indol-3-yl]-4 (1H-indol-3-yl)-1H-pyrrole-2,5-dione monohydrochloride (GF109203X) and 2-[8 [(dimethylamino)methyl]-6,7,8,9-tetrahydropyridol[1,2-a]indol-3-yl]-3-(1 methylindol-3-yl)maleimide (Ro-32-0432), and the extracellular signal-regulated kinase (ERK)1/2 inhibitors 2'-amino-3'-methoxyflavone (PD98059) and 1,4-diamino 2,3-dicyano-1,4-bis(methylthio)butadiene (U0126) all blocked carbachol-induced axonal outgrowth. In addition, down-regulation of ERK1/2 with small interfering RNA abolished the neuritogenic effect of carbachol. These data suggest an involvement of Ca(2+), PKC, and ERK1/2 in carbachol-induced axonal growth. Carbachol indeed increased the release of Ca(2+) from intracellular stores and induced PKC and ERK1/2 activation. Additional experiments showed that PKC, but not Ca(2+), is involved in carbachol-induced ERK1/2 activation. Together, these results show that cholinergic stimulation of prenatal hippocampal pyramidal neurons accelerates axonal growth through the induction of Ca(2+) mobilization and the activation of PKC and especially of ERK1/2. PMID- 19190236 TI - Nonpeptidergic allosteric antagonists differentially bind to the CXCR2 chemokine receptor. AB - The chemokine receptor CXCR2 is involved in different inflammatory diseases, like chronic obstructive pulmonary disease, psoriasis, rheumatoid arthritis, and ulcerative colitis; therefore, it is considered an attractive drug target. Different classes of small CXCR2 antagonists have been developed. In this study, we selected seven CXCR2 antagonists from the diarylurea, imidazolylpyrimide, and thiazolopyrimidine class and studied their mechanisms of action at human CXCR2. All compounds are able to displace (125)I-CXCL8 and inhibit CXCL8-induced beta arrestin2 recruitment. Detailed studies with representatives of each class showed that these compounds displace and antagonize CXCL8, most probably via a noncompetitive, allosteric mechanism. In addition, we radiolabeled the high affinity CXCR2 antagonist SB265610 [1-(2-bromophenyl)-3-(4-cyano-1H-benzo[d] [1,2,3]-triazol-7-yl)urea] and subjected [(3)H]SB265610 to a detailed analysis. The binding of this radioligand was saturable and reversible. Using [(3)H]SB265610, we found that compounds of the different chemical classes bind to distinct binding sites. Hence, the use of a radiolabeled low-molecular weight CXCR2 antagonist serves as a tool to investigate the different binding sites of CXCR2 antagonists in more detail. PMID- 19190237 TI - Smoke, choline acetyltransferase, muscarinic receptors, and fibroblast proliferation in chronic obstructive pulmonary disease. AB - Acetylcholine (ACh), synthesized by choline acetyltransferase (ChAT), and muscarinic M(1), M(2), and M(3) receptors (MRs) are involved in fibroblast proliferation. We evaluated ChAT, MRs, and extracellular signal-regulated kinase (ERK) 1/2 and nuclear factor (NF) kappaB activation in lung fibroblasts from patients with chronic obstructive pulmonary disease (COPD), control smokers, and controls. Human fetal lung fibroblasts (HFL-1) stimulated with interleukin (IL) 1beta, tumor necrosis factor (TNF)-alpha, and cigarette smoke extracts (CSEs) were evaluated for ChAT and MR expression. We tested the effects of ACh on fibroblast proliferation and its ability to bind fibroblasts from patients with COPD, control smokers, controls, and HFL-1 stimulated with IL-1beta, TNF-alpha, and CSE. ChAT, M(1), and M(3) expression and ERK1/2 and NFkappaB activation were increased, whereas M(2) was reduced, in COPD and smoker subjects compared with controls. IL-1beta increased the ChAT and M(3), TNF-alpha down-regulated M(2), and CSE increased ChAT and M(3) expression while down-regulating the expression of M(2) in HFL-1 cells. ACh stimulation increased fibroblast proliferation in patients with COPD, control smokers, and controls, with higher effect in control smokers and patients with COPD and increased HFL-1 proliferation only in CSE treated cells. The binding of ACh was higher in patients with COPD and in control smokers than in controls and in CSE-treated than in IL-1beta- and TNF-alpha stimulated HFL-1 cells. Tiotropium (Spiriva; [1alpha,2beta,4beta,5alpha,7beta-7 hydroxydi-2-thienylacetyl)oxy]-9,9-dimethyl-3-oxa-9-azoniatrcyclo[3.3.1.0(24)], C(19)H(22) NO(4)S(2)Br.H(2)O), gallamine triethiodide (C(19)H(22)N(4)O(2)S.2HCl.H(2)O), telenzepine [4,9-d-dihydro-3-methyl-4-[(4 methyl-1piperazinyl) acetyl]-10H-thieno [3,4-b][1,5]benzodiazepine-10-one dihydrobromide, C(30)H(60)I(3)N(3)O(3)], 4-diphenylacetoxy-N-methylpiperidine, PD098059 [2-(2-amino-3methoxyphenyl)-4H-1benzopyran-4-one, C(16)H(13)NO(3)], and BAY 11-7082 [(E)-3-(4-methylphenylsulfonyl)-2-propenetrile, C(10)H(9)NO(2)C], down-regulated the ACh-induced fibroblast proliferation, promoting the MRs and ERK1/2 and NFkappaB pathways involvement in this phenomenon. These results suggest that cigarette smoke might alter the expression of ChAT and MRs, promoting airway remodeling in COPD and that anticholinergic drugs, including tiotropium, might prevent these events. PMID- 19190238 TI - Prostaglandin E2 derived from cyclooxygenases 1 and 2 mediates intestinal epithelial ion transport stimulated by the activation of protease-activated receptor 2. AB - Proteinase-activated receptor (PAR)(2) is activated by trypsin-like serine proteinases and has been implicated in intestinal inflammation. However, its role in the regulation of intestinal mucosal function remains unclear. Using the intestinal epithelial cell line, SCBN, we have studied the stimulus-secretion coupling mechanisms of PAR(2)-induced epithelial chloride transport, focusing on cyclooxygenase (COX)-1 and COX-2 activities and prostaglandin (PG) E(2) secretion. SCBN monolayers were grown on Snapwell supports, mounted in modified Ussing chambers, and exposed to the activating peptide, SLIGRL-NH(2) (50 microM), to activate PAR(2). Pretreatment with inhibitors of cytosolic PLA(2) (cPLA(2)) (AACOCF3, arachidonyltrifluoromethyl ketone), COX-1 [SC560, 5-(4-chlorophenyl)-1 (4-methoxyphenyl)-3-(trifluoromethyl)-1H-pyrazole], and COX-2 (celecoxib) resulted in a significant concentration-dependent attenuation of PAR(2)-induced changes in short-circuit current. Immunoblot analysis showed a PAR(2)-induced increase in cPLA(2) phosphorylation that was blocked by the mitogen-activated protein kinase kinase inhibitor, PD98059 [2-(2-amino-3methoxyphenyl)-4H 1benzopyran-4-one, C(16)H(13)NO(3)], and the pan-protein kinase C inhibitor, GFX (bisindolylmaleimide). PAR(2) stimulation also resulted in a large increase in the production of PGE(2) as determined by enzyme-linked immunosorbent assay and was also blocked by PD98059 and GFX. Immunofluorescence and immunoblot analysis determined that EP2 and EP4 are expressed at the basolateral membrane of SCBN cells. Through the use of selective inhibitors (EP2, AH6809 [6-isopropoxy-9 oxoxanthene-2-carboxylic acid]; EP4, GW627368X [N-[2[4,9-diethoxy-1-oxo-1,3 dihydro-2H-benzo[f]isoindol-2-yl)phenyl] acetyl]benzene sulphonamide]), it was found that both EP2 and EP4 were involved in mediating the PAR(2)-induced chloride secretory response. We conclude that basolateral PAR(2) activation induces epithelial chloride secretion that is mediated by cPLA(2), COX-1, COX-2, and the subsequent release of PGE(2). The production of PGE(2) results in an autocrine secretory response that is dependent on basolateral EP2 and EP4 receptors. PMID- 19190239 TI - Potential treatment options after first-line chemotherapy for advanced NSCLC: maintenance treatment or early second-line? AB - Although substantial progress has been made in the therapeutic options currently available for patients with advanced non-small cell lung cancer (NSCLC), the overall survival profile remains poor for most patients. One of the strategies currently under investigation with the aim of prolonging survival in NSCLC patients is maintenance treatment with either a chemotherapeutic agent or a molecularly targeted agent after first-line chemotherapy. Moreover, this can consist of drugs included in the induction regimen or other noncrossresistant agents. With the currently available data, maintenance treatment with a different noncrossresistant agent (i.e., an early second-line treatment) is perhaps the most promising strategy. The drug chosen for the early second-line treatment should be a well-tolerated agent, considering that patients have just completed a particularly toxic platinum-based chemotherapy. Extending treatment with targeted agents rather than chemotherapy can provide longer progression-free and overall survival times without increasing toxicity. However, at the moment, only progression-free survival has been shown to be consistently superior with maintenance approaches; the evaluation of survival benefits is warranted before defining this strategy as a possible treatment option. Further studies are warranted to establish the role of maintenance chemotherapy in patients with advanced NSCLC. PMID- 19190240 TI - Ethylene mediates response and tolerance to potassium deprivation in Arabidopsis. AB - Potassium deprivation leads to large reductions in plant growth and yields. How plants sense and transduce the stress signals initiated by potassium deprivation is poorly understood. Both ethylene production and the transcription of genes involved in ethylene biosynthesis increase when plants are deprived of potassium. To elucidate the role of ethylene in low potassium signaling pathways, we used both genetic and chemical approaches. Our results showed that ethylene is important in tolerance to low potassium and for changes in both root hair and primary root growth in Arabidopsis thaliana. We show that ethylene acts upstream of reactive oxygen species in response to potassium deprivation. The expression of High-Affinity K(+) Transporter5 was used as a marker of potassium deprivation and was found to be dependent on ethylene signaling. In the ethylene insensitive2 1 (ein2-1) mutant, the ethylene-mediated low potassium responses were not completely eliminated, suggesting that some potassium deprivation-induced responses are either ethylene independent or EIN2 independent. Ethylene signaling is a component of the plant's response to low potassium that stimulates the production of reactive oxygen species and is important for changes in root morphology and whole plant tolerance to low potassium conditions. PMID- 19190241 TI - Genetic regulation of meiotic cross-overs between related genomes in Brassica napus haploids and hybrids. AB - Although the genetic regulation of recombination in allopolyploid species plays a pivotal role in evolution and plant breeding, it has received little recent attention, except in wheat (Triticum aestivum). PrBn is the main locus that determines the number of nonhomologous associations during meiosis of microspore cultured Brassica napus haploids (AC; 19 chromosomes). In this study, we examined the role played by PrBn in recombination. We generated two haploid x euploid populations using two B. napus haploids with differing PrBn (and interacting genes) activity. We analyzed molecular marker transmission in these two populations to compare genetic changes, which have arisen during meiosis. We found that cross-over number in these two genotypes was significantly different but that cross-overs between nonhomologous chromosomes showed roughly the same distribution pattern. We then examined genetic recombination along a pair of A chromosomes during meiosis of B. rapa x B. napus AAC and AACC hybrids that were produced with the same two B. napus genotypes. We observed significant genotypic variation in cross-over rates between the two AAC hybrids but no difference between the two AACC hybrids. Overall, our results show that PrBn changes the rate of recombination between nonhomologous chromosomes during meiosis of B. napus haploids and also affects homologous recombination with an effect that depends on plant karyotype. PMID- 19190242 TI - Cdc42-dependent leading edge coordination is essential for interstitial dendritic cell migration. AB - Mature dendritic cells (DCs) moving from the skin to the lymph node are a prototypic example of rapidly migrating amoeboid leukocytes. Interstitial DC migration is directionally guided by chemokines, but independent of specific adhesive interactions with the tissue as well as pericellular proteolysis. Instead, the protrusive flow of the actin cytoskeleton directly drives a basal mode of locomotion that is occasionally supported by actomyosin contractions at the trailing edge to propel the cell's rigid nucleus. We here delete the small GTPase Cdc42 in DCs and find that actin flow and actomyosin contraction are still initiated in response to chemotactic cues. Accordingly, the cells are able to polarize and form protrusions. However, in the absence of Cdc42 the protrusions are temporally and spatially dysregulated, which leads to impaired leading edge coordination. Although this defect still allows the cells to move on 2 dimensional surfaces, their in vivo motility is completely abrogated. We show that this difference is entirely caused by the geometric complexity of the environment, as multiple competing protrusions lead to instantaneous entanglement within 3-dimensional extracellular matrix scaffolds. This demonstrates that the decisive factor for migrating DCs is not specific interaction with the extracellular environment, but adequate coordination of cytoskeletal flow. PMID- 19190243 TI - Nutlin-3 up-regulates the expression of Notch1 in both myeloid and lymphoid leukemic cells, as part of a negative feedback antiapoptotic mechanism. AB - The small molecule inhibitor of the MDM2/p53 interaction Nutlin-3 significantly up-regulated the steady-state mRNA and protein levels of Notch1 in TP53(wild type) (OCI, SKW6.4) but not in TP53(deleted) (HL-60) or TP53(mutated) (BJAB) leukemic cell lines. A direct demonstration that NOTCH1 was a transcriptional target of p53 in leukemic cells was obtained in experiments carried out with siRNA for p53. Moreover, inhibition of Notch1 expression using Notch1-specific siRNA significantly increased cytotoxicity in TP53(wild-type) leukemic cells. Of note, Nutlin-3 up-regulated Notch1 expression also in primary TP53(wild-type) B chronic lymphocytic leukemia (B-CLL) cells and the combined use of Nutlin-3 plus pharmacological gamma-secretase inhibitors of the Notch signaling showed a synergistic cytotoxicity in both TP53(wild-type) leukemic cell lines and primary B-CLL cells. A potential drawback of gamma-secretase inhibitors was their ability to enhance osteoclastic maturation of normal circulating preosteoclasts induced by RANKL + M-CSF. Notwithstanding, Nutlin-3 completely suppressed osteoclastogenesis irrespective of the presence of gamma-secretase inhibitors. Taken together, these data indicate that the p53-dependent up-regulation of Notch1 in response to Nutlin-3 represents an antiapoptotic feedback mechanism able to restrain the potential therapeutic efficacy of Nutlin-3 in hematologic malignancies. Therefore, therapeutic combinations of Nutlin-3 + gamma-secretase inhibitors might potentiate the cytotoxicity of Nutlin-3 in p53(wild-type) leukemic cells. PMID- 19190244 TI - p85beta phosphoinositide 3-kinase regulates CD28 coreceptor function. AB - CD28 is a receptor expressed on T cells that regulates their differentiation after antigen stimulation to long-term-survival memory T cells. CD28 enhances T cell receptor signals and reduces expression of CBL ubiquitin ligases, which negatively control T-cell activation. In the absence of CD28 ligation during the primary stimulation, CBL levels remain high and T cells fail to mount an efficient secondary response. CD28 associates with p85alpha, one of the regulatory subunits of phosphoinositide-3-kinase (PI3K), but the relevance of this interaction is debated. We examined here the contribution of the other ubiquitous PI3K regulatory subunit, p85beta, in CD28 function. We describe that p85beta bound to CD28 and to CBL with greater affinity than p85alpha. Moreover, deletion of p85beta impaired CD28-induced intracellular events, including c-CBL and CBL-b down-regulation as well as PI3K pathway activation. This resulted in defective differentiation of activated T cells, which failed to exhibit an efficient secondary immune response. Considering that p85beta-deficient T cells fail in recall responses and that p85beta binds to and regulates CD28 signals, the presented observations suggest the involvement of p85beta in CD28-mediated activation and differentiation of antigen-stimulated T cells. PMID- 19190245 TI - Cytoskeletal protein 4.1R negatively regulates T-cell activation by inhibiting the phosphorylation of LAT. AB - Protein 4.1R (4.1R) was first identified in red cells where it plays an important role in maintaining mechanical stability of red cell membrane. 4.1R has also been shown to be expressed in T cells, but its function has been unclear. In the present study, we use 4.1R-deficient mice to explore the role of 4.1R in T cells. We show that 4.1R is recruited to the immunologic synapse after T cell-antigen receptor (TCR) stimulation. We show further that CD4+ T cells of 4.1R-/- mice are hyperactivated and that they displayed hyperproliferation and increased production of interleukin-2 (IL-2) and interferon gamma (IFNgamma). The hyperactivation results from enhanced phosphorylation of LAT and its downstream signaling molecule ERK. The 4.1R exerts its effect by binding directly to LAT, and thereby inhibiting its phosphorylation by ZAP-70. Moreover, mice deficient in 4.1R display an elevated humoral response to immunization with T cell-dependent antigen. Thus, we have defined a hitherto unrecognized role for 4.1R in negatively regulating T-cell activation by modulating intracellular signal transduction. PMID- 19190246 TI - Loss of pleckstrin defines a novel pathway for PKC-mediated exocytosis. AB - Pleckstrin, the platelet and leukocyte C kinase substrate, is a prominent substrate of PKC in platelets, monocytes, macrophages, lymphocytes, and granulocytes. Pleckstrin accounts for 1% of the total protein in these cells, but it is best known for containing the 2 prototypic Pleckstrin homology, or PH, domains. Overexpressed pleckstrin can affect polyphosphoinositide second messenger-based signaling events; however, its true in vivo role has been unknown. Here, we describe mice containing a null mutation within the pleckstrin gene. Platelets lacking pleckstrin exhibit a marked defect in exocytosis of delta and alpha granules, alphaIIbbeta3 activation, actin assembly, and aggregation after exposure to the PKC stimulant, PMA. Pleckstrin-null platelets aggregate normally in response to thrombin, but they fail to aggregate in response to thrombin in the presence of PI3K inhibitors, suggesting that a PI3K-dependent signaling pathway compensates for the loss of pleckstrin. Although pleckstrin null platelets merged their granules in response to stimulation of PKC, they failed to empty their contents into the open canalicular system. This might be attributable to impaired actin assembly present in cells lacking pleckstrin. These data show that pleckstrin regulates the fusion of granules to the cell membrane and is an essential component of PKC-mediated exocytosis. PMID- 19190248 TI - Adverse events among 2408 unrelated donors of peripheral blood stem cells: results of a prospective trial from the National Marrow Donor Program. AB - Limited data are available describing donor adverse events (AEs) associated with filgrastim mobilized peripheral blood stem cell (PBSC) collections in unrelated volunteers. We report results in 2408 unrelated PBSC donors prospectively evaluated by the National Marrow Donor Program (NMDP) between 1999 and 2004. Female donors had higher rates of AEs, requiring central line placement more often (17% vs 4%, P< .001), experiencing more apheresis-related AEs (20% vs 7%, P< .001), more bone pain (odds ratio [OR]=1.49), and higher rates of grades II-IV and III-IV CALGB AEs (OR=2.22 and 2.32). Obese donors experienced more bone pain (obese vs normal, OR=1.73) and heavy donors had higher rates of CALGB toxicities (>95 kg vs <70 kg, OR=1.49). Six percent of donors experienced grade III-IV CALGB toxicities and 0.6% experienced toxicities that were considered serious and unexpected. Complete recovery is universal, however, and no late AEs attributable to donation have been identified. In conclusion, PBSC collection in unrelated donors is generally safe, but nearly all donors will experience bone pain, 1 in 4 will have significant headache, nausea, or citrate toxicity, and a small percentage will experience serious short-term adverse events. In addition, women and larger donors are at higher risk for donation-related AEs. PMID- 19190247 TI - The novel plant-derived agent silvestrol has B-cell selective activity in chronic lymphocytic leukemia and acute lymphoblastic leukemia in vitro and in vivo. AB - Therapeutic options for advanced B-cell acute lymphoblastic leukemia (ALL) and chronic lymphocytic leukemia (CLL) are limited. Available treatments can also deplete T lymphocytes, leaving patients at risk of life-threatening infections. In the National Cancer Institute cell line screen, the structurally unique natural product silvestrol produces an unusual pattern of cytotoxicity that suggests activity in leukemia and selectivity for B cells. We investigated silvestrol efficacy using primary human B-leukemia cells, established B-leukemia cell lines, and animal models. In CLL cells, silvestrol LC(50) (concentration lethal to 50%) is 6.9 nM at 72 hours. At this concentration, there is no difference in sensitivity of cells from patients with or without the del(17p13.1) abnormality. In isolated cells and whole blood, silvestrol is more cytotoxic toward B cells than T cells. Silvestrol causes early reduction in Mcl-1 expression due to translational inhibition with subsequent mitochondrial damage, as evidenced by reactive oxygen species generation and membrane depolarization. In vivo, silvestrol causes significant B-cell reduction in Emu-Tcl-1 transgenic mice and significantly extends survival of 697 xenograft severe combined immunodeficient (SCID) mice without discernible toxicity. These data indicate silvestrol has efficacy against B cells in vitro and in vivo and identify translational inhibition as a potential therapeutic target in B-cell leukemias. PMID- 19190249 TI - Green tea polyphenols block the anticancer effects of bortezomib and other boronic acid-based proteasome inhibitors. AB - The anticancer potency of green tea and its individual components is being intensely investigated, and some cancer patients already self-medicate with this "miracle herb" in hopes of augmenting the anticancer outcome of their chemotherapy. Bortezomib (BZM) is a proteasome inhibitor in clinical use for multiple myeloma. Here, we investigated whether the combination of these compounds would yield increased antitumor efficacy in multiple myeloma and glioblastoma cell lines in vitro and in vivo. Unexpectedly, we discovered that various green tea constituents, in particular (-)-epigallocatechin gallate (EGCG) and other polyphenols with 1,2-benzenediol moieties, effectively prevented tumor cell death induced by BZM in vitro and in vivo. This pronounced antagonistic function of EGCG was evident only with boronic acid-based proteasome inhibitors (BZM, MG-262, PS-IX), but not with several non-boronic acid proteasome inhibitors (MG-132, PS-I, nelfinavir). EGCG directly reacted with BZM and blocked its proteasome inhibitory function; as a consequence, BZM could not trigger endoplasmic reticulum stress or caspase-7 activation, and did not induce tumor cell death. Taken together, our results indicate that green tea polyphenols may have the potential to negate the therapeutic efficacy of BZM and suggest that consumption of green tea products may be contraindicated during cancer therapy with BZM. PMID- 19190250 TI - Subcortical alterations in tissue microstructure adjacent to focal cortical dysplasia: detection at diffusion-tensor MR imaging by using magnetoencephalographic dipole cluster localization. AB - PURPOSE: To determine whether changes at diffusion-tensor magnetic resonance (MR) imaging were present in children with intractable epilepsy and focal cortical dysplasia (FCD) in (a) subcortical white matter subjacent to MR imaging-visible areas of FCD, (b) subcortical white matter beyond the MR imaging-visible abnormality but subjacent to a magnetoencephalographic (MEG) dipole cluster, and (c) deep white matter tracts. MATERIALS AND METHODS: The study protocol had institutional research ethics board approval, and written informed consent was obtained. Fifteen children with FCD and intractable epilepsy (mean age, 11.6 years; range, 3.6-18.3 years) underwent diffusion-tensor MR imaging and MEG. Regions of interest were placed in (a) the subcortical white matter subjacent to the MR imaging-visible abnormality, as well as the contralateral side; (b) the subcortical white matter beyond the MR imaging-visible abnormality but subjacent to a MEG dipole cluster, as well as the contralateral side; and (c) deep white matter tracts projecting to or from the MR imaging-visible FCD, as well as the contralateral side. Fractional anisotropy (FA), mean diffusivity, and eigenvalues (lambda(1), lambda(2), lambda(3)) were evaluated. RESULTS: Eleven of 15 children had MEG dipole clusters, and four children had MEG scatter. There were significant differences in FA, mean diffusivity, lambda(2), and lambda(3) of the subcortical white matter subjacent to the MR imaging-visible FCD (P < .001 for all), as well as that beyond the MR imaging-visible FCD but subjacent to a MEG dipole cluster (P = .001, P = .036, P < .001, and P = .002, respectively), compared with the contralateral side. There were also significant differences in FA (P < .001), mean diffusivity (P = .008), lambda(2) (P < .001), and lambda(3) (P = .001) of the deep white matter tracts projecting to or from the MR imaging visible FCD compared with the contralateral side. CONCLUSION: With use of MEG dipole clusters to localize the epileptogenic zone, diffusion-tensor imaging can help identify alterations in tissue microstructure beyond the MR imaging-visible FCD. PMID- 19190251 TI - Difference in tracer delay-induced effect among deconvolution algorithms in CT perfusion analysis: quantitative evaluation with digital phantoms. AB - Institutional review board approval and informed consent were obtained. The purpose was to evaluate the differences in tracer delay-induced effects of various deconvolution algorithms for computed tomographic (CT) perfusion imaging by using digital phantoms created from actual source data. Three methods of singular value decomposition (SVD) were evaluated. For standard SVD (sSVD), the delays induced significant errors in cerebral blood flow and mean transit time. In contrast, for block-circulant SVD (bSVD), these values remained virtually unchanged, whereas for delay-corrected SVD (dSVD), mild changes were observed. bSVD was superior to sSVD and dSVD for avoiding the tracer delay-induced effects in CT perfusion imaging. PMID- 19190252 TI - Detection of epidermal thickening in GJB2 carriers with epidermal US. AB - PURPOSE: To measure epidermal thickness by using skin ultrasonography (US) in a series of healthy control subjects and obligate carriers for the worldwide most frequent form of congenital hearing loss owing to the mutated alleles of the connexin 26 gene (GJB2). MATERIALS AND METHODS: The patent for the protocol, coupled with a new sonographic probe specifically designed to analyze epidermal thickness and a dedicated algorithm to classify individuals in groups, is pending. Institutional ethics committee approval and patient consent were obtained. After a preliminary study in 23 subjects aimed to define the best body site and instrument and protocol for US, a total of 303 individuals (237 healthy subjects, 51 carriers, and 15 homozygotes) were tested at midline forehead by using a linear large-band probe with a frequency ranging from 6 to 15 MHz to determine epidermal thickness. Variance and linear regression analyses were performed. Regression coefficients were then used to obtain measurements of thickness corrected for age and sex. RESULTS: GJB2 obligate carriers had a significant increase in epidermal thickness compared with control subjects. GJB2 status explains about 50.0% of this variability, whereas an additional 25.0% is explained by sex and age. Results led to the development of a possible screening protocol with a 98.0% sensitivity and 92.8% specificity in subjects aged 2080 years, with a likelihood ratio of a positive test of 14:1. Even better results (100% sensitivity and 98.9% specificity) were obtained in an analysis of people of only reproductive age. CONCLUSION: Epidermal thickening in the white population owing to GJB2 carrier status can be detected by using US. This measurement could provide a simple, noninvasive, rapid, and sensitive test for carrier screening. PMID- 19190253 TI - Maternal years of schooling but not academic skills is independently associated with infant-feeding practices in a cohort of rural Guatemalan women. AB - The effects of maternal academic skills on infant-feeding practices are not clear. From 1996 to 1999, the authors collected information on infant-feeding practices from birth on infants born to 279 mothers from 4 rural villages in Guatemala. They examined associations between maternal academic skills and indicators for the initiation of exclusive breastfeeding (EBF) and timely introduction of complementary foods (CF). Mothers in the highest category of academic skills had greater odds of initiating EBF, but this association failed to remain significant after adjusting for schooling. Compared with mothers with < 1 year of school, mothers with > 3 to 6 years of school had greater odds of introducing CF early, while mothers with >or= 1 to or =18 years from three regions of Alaska completed questions on safety behaviours as part of a comprehensive health and lifestyle computer assisted self-administered questionnaire. RESULTS: Most participants reported never driving after drinking (94.1%) or riding with a driver who had been drinking (91.3%). Fewer (74.1%) participants reported using a seatbelt always or almost always when riding in a vehicle. Only about half (55%) always kept to the speed limit when driving or used a personal flotation device when boating (51.5%). Even fewer (20.5%) reported using a helmet when riding on off-road vehicles, including four-wheelers and snowmobiles. Factors identified among those least likely to use safety devices and practise good safety behaviours are: younger age, lower household income and education, non-married, speaking only English at home, and a self-reported health status of poor to fair (p<0.05). CONCLUSIONS: Recommendations for future injury prevention efforts in this population are to increase use of personal flotation devices while boating and address the underuse of helmets with off-road vehicles. Limited prevention resources should be used to target those who engage in risky behaviours to maximise programme impact. PMID- 19190274 TI - Injuries from interpersonal violence presenting to a rural health center in Western Kenya: characteristics and correlates. AB - OBJECTIVE: To define the scope of injury due to interpersonal violence in a medium-sized town in Western Kenya. DESIGN: Prospective, cross-sectional data collection and analysis. SETTING/SUBJECTS: Data were prospectively collected on all injured patients (n = 562) presenting to a health center in Western Kenya, 2002-2004. Age, gender, type, and severity of injury, relationship to assailant, disposition, and clinician's suspicion of alcohol use were recorded. MAIN OUTCOME MEASURES: Number of injuries due to interpersonal violence; correlation of gender, alcohol use, relationship to assailant, and type of injury. RESULTS: Interpersonal violence caused 43% of all injuries. Men and women were equally likely to suffer violent injuries (42% vs 45%); however, women were more likely to suffer injury from domestic violence (4.7% vs 7.0%) and sexual assault (0% vs 3.5%). Men and women were equally likely to know their assailant. Women were more likely to be injured by a spouse/partner (19% vs 1.3%), whereas men were more likely to be injured by an acquaintance (29% vs 16%). Alcohol use was more often suspected for victims of violent, as opposed to unintentional, injury (45% vs 16%). Men with violent injuries were more likely than women to be suspected of having used alcohol (51% vs 35%). CONCLUSIONS: Interpersonal violence is a leading cause of injury in Western Kenya. Although men and women are equally likely to be assaulted, women are more likely to be injured by a spouse, and men by an acquaintance. Alcohol use is common among those who suffer violent injuries in this population. PMID- 19190275 TI - Descriptive epidemiological study of sites of suicide jumps in Taipei, Taiwan. AB - OBJECTIVE: To identify locations to target for suicide preventive measures to reduce suicide by jumping in Taipei city, Taiwan. METHOD: A descriptive study of suicide by jumping from a height was conducted in Taipei, 2002-2005. Information on sites from which suicide jumps occurred was obtained from death certificates as well as ambulance, medical and newspaper records. RESULTS: Of the 210 jumping suicide deaths with sufficient information on the site of the jump, private residential buildings comprised the highest proportion (67%) of all jumping sites, followed by business office buildings (13%), hospitals (8%) and shopping malls (5%). However, the number of jumping suicides per 1000 sites was highest for hospitals (307 per 1000 hospitals) and shopping malls (275 per 1000 shopping malls) compared with 2 per 1000 for residential buildings. Women and older people who died by jumping were more likely to have jumped from buildings where they lived, compared with men and other age groups. CONCLUSION: Although two-thirds of suicides occurred from residential buildings, the sites with the greatest number of jumps per location were hospitals and shopping malls, making them the most cost-effective focus for preventive efforts. PMID- 19190276 TI - Asphyxial games or "the choking game": a potentially fatal risk behaviour. AB - OBJECTIVES: To determine the prevalence of knowledge about and participation in asphyxial games, sometimes called "the choking game", and how best to raise awareness of this risk-taking behaviour and provide preventive education. DESIGN: Questionnaire; collaborative research model; lay advocacy group/university researchers. SETTING: 8 middle and high schools in Texas (six) and Ontario (two). A recent death from playing the choking game had occurred in one Texas school, and two other fatalities had occurred within the state. SUBJECTS: Students in grades 4-12, aged 9-18 years. INTERVENTION: None. OUTCOME MEASURES: None. RESULTS: Of 2762 surveys distributed, 2504 (90.7%) were completed. The mean (SD) age of the responders was 13.7 (2.2) years. 68% of children had heard about the game, 45% knew somebody who played it, and 6.6% had tried it, 93.9% of those with someone else. Forty percent of children perceived no risk. Information that playing the game could result in death or brain damage was reported as most likely to influence behaviour. The most respected source of a preventive education message was parents for pre-adolescents (43%) or victim/victim's family (36%) for older adolescents. CONCLUSIONS: Knowledge of and participation in self asphyxial behaviour is not unusual among schoolchildren. The age of the child probably determines the best source (parents or victim/victim's family) of preventive education. PMID- 19190277 TI - Once bitten, twice shy? Medically-attended injuries can sensitise parents to children's risk of injuries on playgrounds. AB - OBJECTIVE: To compare risk perceptions of parents whose child sustained a medically attended playground injury (cases) with those of parents whose child had not (controls) to address two questions. Does having a child experience a medically attended injury: (1) sensitise parents to children's injury vulnerability and severity; (2) influence parents' appraisal of the injury mechanism (child's behaviour), attributions for injuries or beliefs about strategies for prevention? METHOD: Each case-control parent dyad was assigned to one of two conditions: (1) being presented with 10 common injury-risk playground behaviours specific to the equipment on which their child had been hurt, and asked to appraise injury vulnerability and severity; or (2) being presented with scenarios about playground injuries that varied in severity but were all based on the same child behaviour, and asked questions about this behaviour, attributions for injury and strategies for prevention. RESULTS: The results support the occurrence of a sensitisation process. Compared with control parents, case parents showed higher ratings of injury severity and children's vulnerability to injury, made fewer attributions for injuries to bad luck, and endorsed a greater diversity of prevention strategies, including parent (closer supervision), child (teaching rules about safe play on playgrounds) and environmental (modifications to playgrounds). CONCLUSIONS: A child's medically attended injury can create a "teachable moment" for the parent. Linking injury-prevention programming to this teachable moment may increase the likelihood of evoking changes in parental supervisory behaviour and their setting of rules limiting their child's risk behaviours to reduce the occurrence of childhood injury. PMID- 19190278 TI - Distribution of road traffic deaths by road user group: a global comparison. AB - BACKGROUND: Road traffic deaths are a major global health and development problem. An understanding of the existing burden of road traffic deaths in the population is necessary for developing effective interventions. OBJECTIVE: To outline systematically the global distribution of road traffic deaths by road user groups (pedestrians, bicyclists, motorcyclists, motorised four-wheeler occupants). METHODS: Comprehensive searches of PubMed, Google, Google Scholar, TransportLink, grey literature and reference lists and communication with experts from international organisations and country-level institutions were conducted to identify eligible studies and data sources. All data sources that provided a breakdown of road traffic deaths by road user group at the national or sub national level were eligible for inclusion. A breakdown of road traffic deaths by road user group was constructed for 14 epidemiologically defined World Health Organization (WHO) sub-regions in addition to World Bank income categories. In addition, the total number of road traffic fatalities by road user group in low income, middle-income and high-income countries was estimated. RESULTS: The breakdown of road traffic deaths by road user group varies dramatically across epidemiological WHO sub-regions. The magnitude of pedestrian fatalities ranges from more than half in African sub-region AfrE (55%) to 15% or less in AmrA or EurA. The distribution also varies across low-income, middle-income and high income countries. 45% of road traffic fatalities in low-income countries are among pedestrians, whereas an estimated 29% in middle-income and 18% in high income countries are among pedestrians. The burden of road traffic injuries on vulnerable road users differs substantially across income levels. An estimated total of 227,835 pedestrians die in low-income countries, as opposed to 161,501 in middle-income countries and 22,500 in high-income countries each year. CONCLUSIONS: Ameliorating road safety requires the implementation of context specific solutions. This review of the road traffic injury literature provides strong evidence that the distribution of road traffic fatalities varies dramatically across different parts of the world. Therefore, context-appropriate and effective prevention strategies that protect the particular at-risk road user groups should be carefully investigated. PMID- 19190279 TI - Accuracy of external cause-of-injury coding in hospital records. AB - OBJECTIVE: To appraise the published evidence regarding the accuracy of external cause-of-injury codes in hospital records. DESIGN: Systematic review. DATA SOURCES: Electronic databases searched included PubMed, PubMed Central, Medline, CINAHL, Academic Search Elite, Proquest Health and Medical Complete, and Google Scholar. Snowballing strategies were used by searching the bibliographies of retrieved references to identify relevant associated articles. SELECTION CRITERIA: Studies were included in the review if they assessed the accuracy of external cause-of-injury coding in hospital records via a recoding methodology. METHODS: The papers identified through the search were independently screened by two authors for inclusion. Because of heterogeneity between studies, meta analysis was not performed. RESULTS: Very limited research on the accuracy of external cause coding for injury-related hospitalisation using medical record review and recoding methodologies has been conducted, with only five studies matching the selection criteria. The accuracy of external cause coding using ICD 9-CM ranged from approximately 64% when exact code agreement was examined to approximately 85% when agreement for broader groups of codes was examined. CONCLUSIONS: Although broad external cause groupings coded in ICD-9-CM can be used with some confidence, researchers should exercise caution for very specific codes until further research is conducted to validate these data. As all previous studies have been conducted using ICD-9-CM, research is needed to quantify the accuracy of coding using ICD-10-AM, and validate the use of these data for injury surveillance purposes. PMID- 19190280 TI - MENTOR-VIP: helping develop key skills for injury and violence prevention. PMID- 19190281 TI - The role of psychology in injury prevention efforts. PMID- 19190284 TI - Telling stories. PMID- 19190286 TI - Teaching evidence-based psychiatry to residents and fellows: developing the curriculum. PMID- 19190288 TI - Where there's smoke, can there be fire? PMID- 19190289 TI - The teaching of psychiatry to non-psychiatrists: the patient as a person. PMID- 19190290 TI - Evidence-based medicine in the education of psychiatrists. AB - OBJECTIVE: Evidence-based medicine has an important place in the teaching and practice of psychiatry. Attempts to teach evidence-based medicine skills can be weakened by conceptual confusions feeding a false polarization between traditional clinical skills and evidence-based medicine. METHODS: The author develops a broader conception of clinical expertise consisting of three tasks, clarifies the role of evidence-based medicine, and explores the implications for teaching and practice. RESULTS: Evidence-based medicine is an essential tool that enables clinicians to assess causal explanations in etiology, risk, or prognosis, or to compare treatments. CONCLUSION: An explicit and inclusive conceptualization of clinical skills can provide a framework to implement and assess curricular efforts to teach evidence-based medicine. PMID- 19190291 TI - Teaching evidence-based psychiatry: integrating and aligning the formal and hidden curricula. AB - OBJECTIVE: The authors argue that adopting evidence-based psychiatry will require a paradigm shift in the training of psychiatry residents, and offer some suggestions for how this transformation might be achieved. METHODS: The authors review the growing literature that addresses how best to teach evidence-based medicine and highlight several examples of innovative instructional and assessment methods. RESULTS: Little is known about how best to instill among residents the attitudes, knowledge, skills, and behaviors that are necessary to practice evidence-based psychiatry. However, there are indications that the integration of evidence-based medicine instruction into routine clinical care and the alignment of the "hidden curriculum" with evidence-based practice are important. CONCLUSION: A whole-program approach may be necessary to create the conditions required in postgraduate training to produce evidence-based psychiatrists. PMID- 19190292 TI - Guided mentorship in evidence-based medicine for psychiatry: a pilot cohort study supporting a promising method of real-time clinical instruction. AB - OBJECTIVE: Evidence-based medicine has been promoted to enhance clinical decision making and outcomes in psychiatry. Residency training programs do not routinely provide instruction in evidence-based medicine. Where instruction exists, it tends to occur in classroom settings divorced from the clinical decision-making process and is focused narrowly on appraisal of evidence quality. The goal of this pilot study was to develop and evaluate the promise of a method of "hands on" instruction in evidence-based medicine done in real clinical time. METHODS: A modularized curriculum to promote decisionmaking strategies using evidence-based medicine during the course of actual patient care was delivered by an attending physician mentoring a small team on the inpatient and consultation-liaison psychiatry services at Stanford. A staggered cohort of 24 consecutive trainees was followed between August and January 2007. Measures of trainees' skills in evidence-based medicine were assessed before and after mentoring. A blinded grader scored each inventory according to an explicit, predefined rubric. Demonstrated proficiency in delivery in each of the core skills of evidence-based medicine was assessed as a secondary outcome measure via the attending physician's unblinded subjective evaluation of trainee performance. Subjective descriptions of the experience were obtained via review of trainees' evaluations. RESULTS: Postmeasures of knowledge and skills in evidence-based medicine increased significantly relative to baseline. The Cohen's d effect size was large and clinically meaningful. The majority of trainees were able to demonstrate adequate proficiency of skills by attending subjective evaluation. Trainees' subjective experiences overall were positive. CONCLUSION: Guided mentoring in evidence-based medicine appears promising for further study. PMID- 19190293 TI - Evaluation of an evidence-based tobacco treatment curriculum for psychiatry residency training programs. AB - OBJECTIVE: Smokers with mental illness and addictive disorders account for nearly one in two cigarettes sold in the United States and are at high risk for smoking related deaths and disability. Psychiatry residency programs provide a unique arena for disseminating tobacco treatment guidelines, influencing professional norms, and increasing access to tobacco cessation services among smokers with mental illness. The current study evaluated the Rx for Change in Psychiatry curriculum, developed for psychiatry residency programs and focused on identifying and treating tobacco dependence among individuals with mental illness. METHODS: The 4-hour curriculum emphasized evidence-based, patient oriented cessation treatments relevant for all tobacco users, including those not yet ready to quit. The curriculum was informed by comprehensive literature review, consultation with an expert advisory group, faculty interviews, and a focus group with psychiatry residents. This study reports on evaluation of the curriculum in 2005-2006, using a quasi-experimental design, with 55 residents in three psychiatry residency training programs in Northern California. RESULTS: The curriculum was associated with improvements in psychiatry residents' knowledge, attitudes, confidence, and counseling behaviors for treating tobacco use among their patients, with initial changes from pre- to posttraining sustained at 3 months' follow-up. Residents' self-reported changes in treating patients' tobacco use were substantiated through systematic chart review. CONCLUSION: The evidence based Rx for Change in Psychiatry curriculum is offered as a model tobacco treatment curriculum that can be implemented in psychiatry residency training programs and disseminated widely, thereby effectively reaching a vulnerable and costly population of smokers. PMID- 19190294 TI - Peer-mentored research development meeting: a model for successful peer mentoring among junior level researchers. AB - OBJECTIVE: This report describes a model for the development, process, and tracking methods of a Peer-mentored Research Development Meeting (PRDM), an interdisciplinary peer mentoring program. The program was initiated in 2004 by a group of postdoctoral scholars and junior faculty from the Schools of the Health Sciences at the University of Pittsburgh. METHOD: From February 2004 through February 2006, PRDM's first five members tracked and documented their research activity (e.g., manuscripts, grants) every 4 months. The defining features of PRDM are adherence to a structured frequency and format for meetings, systematic tracking and evaluation of research development activities, and maintenance of ongoing relationships with senior mentors. RESULTS: During the 24-month data collection period, members were involved in 91 research development projects including grant applications, journal article manuscripts, book chapters, and conference abstracts. Members' productivity increased during the 24-month period, as did the efficiency and focus of the completed projects. CONCLUSION: Members increased the efficiency and focus of their research development activities during the study period. Structured peer-mentoring groups have the potential to enhance research productivity among junior investigators in research intensive environments. PMID- 19190295 TI - Can admissions interviews predict performance in residency? AB - OBJECTIVE: The authors aim to determine whether admission interviews predict performance in residency. METHODS: The authors determined whether interview and other admission data were correlated with performance during postgraduate years 2 4 and with remaining in the residency in 544 residents enrolled in a single psychiatry residency program between 1963 and 2004. RESULTS: Considered together, admissions data predicted 13% of the variance in performance ratings in postgraduate year 2 (PGY-2) and 5% in PGY-4. Interview scores were moderately correlated with performance ratings in PGY-2, modestly correlated with performance in PGY-3, and not correlated with performance ratings in PGY-4. Letters of reference were moderately correlated with performance ratings in PGY-2 and modestly correlated with performance in PGY-3 and PGY-4. In PGY-2, interview scores differentiated between the top quartile of performance and the other three quartiles, while letters of reference differentiated performance in the top and bottom quartiles from the middle quartiles. Numerical differences among groups were not great enough to be practically useful, and no variables predicted which residents would leave the program before completing it. CONCLUSION: As they are currently conducted, application interviews do not have sufficient power to predict performance during residency. Letters of reference may be useful to the extent that they reflect personal experience with the applicant, but differences in ratings of these letters are not great enough to base admission decisions on them. As it is currently performed, the interview process may be more useful as a means of interesting applicants in the program than of evaluating their potential for success in the residency. PMID- 19190296 TI - Teaching psychiatry in primary care residencies: do training directors of primary care and psychiatry see eye to eye? AB - OBJECTIVE: This study compares the views of psychiatry residency training directors about psychiatry and mental health training in the primary care programs in their institutions with those of the primary care residency training directors. METHODS: A 16-item questionnaire surveying specific areas of training and perceived adequacy of current teaching was distributed to 1,544 U.S. primary care and psychiatry program directors. RESULTS: The response rate was 53%. Among psychiatry training directors, 85% responded that psychiatry training in their primary care programs was minimal to suboptimal, while 68% of family practice training directors responded that their psychiatry training was optimal to extensive. Among psychiatry training directors, 89% were dissatisfied with the psychiatry training in their primary care programs, and only 8% were satisfied. In contrast, almost half of primary care training directors were satisfied. However, within the primary care programs, there was a marked difference between family practice (majority satisfied) and the rest (internal medicine, obstetrics and gynecology, pediatrics, mostly unsatisfied). All primary care and psychiatry training directors agreed that most basic psychiatric skills and diagnoses were taught in the primary care programs. For all skills and syndromes examined, psychiatry training directors consistently and significantly rated the training to be less adequate than did primary care training directors. There was general agreement that primary care physicians should be able to treat most uncomplicated cases in patients with psychiatric disorders, and some but not other psychiatric conditions. CONCLUSION: Psychiatry and primary care training directors, except in family practice, generally agree that psychiatry training in primary care programs is inadequate and should be significantly enhanced. There should be more communication between psychiatry and primary care training programs for optimal curriculum development. PMID- 19190297 TI - A novel network for mentoring family physicians on mental health issues using E mail. AB - OBJECTIVE: Family practitioners are significant providers of mental health care and routinely report difficulty acquiring timely support in this area. The Collaborative Mental Health Care Network assembled groups of family practitioners and provided them with mental health practitioner mentors. This article addresses communication in the Network, its effect on family practitioners, and the role e mail plays. METHODS: This descriptive study utilizes two sources of data: a quality assurance survey administered to family practitioners in the Network and a sampling of e-mail correspondence between family practitioners and mental health and addiction mentors, examined qualitatively. RESULTS: Family practitioners in the Network requested consultation on pharmacotherapy (53%), psychotherapy (34%), treatment review (27%), and diagnosis (24%). Satisfaction with the Network was high, with 88% of family practitioners reporting an improvement in ability to provide mental health care. E-mail analysis suggests that mentors convey information directly and indirectly and that a knowledge hierarchy, but not a power hierarchy, develops. The trusted relationship between the mentee and mentor is an important context for effective education. CONCLUSION: This model of mentoring is highly satisfactory to family practitioners and correlates with increased confidence in caring for patients with mental health issues. E-mail is a promising strategy for effective feedback and support between family practitioners and specialists. PMID- 19190298 TI - Positive attitude change toward psychiatry in pharmacy students following an active learning psychopharmacology course. AB - OBJECTIVE: Psychiatric care in many rural communities has been demonstrated to be less adequate compared with urban environments partially because of attitudes and stigmatization issues. Educated pharmacists with professional attitudes can have a major impact in helping mental health patients receive more accurate diagnostic assessments and safe and effective treatment. This report demonstrates the effects of a specialized elective course in psychopharmacology on doctor of pharmacy students' attitudes toward psychiatric disorders and their treatment. METHODS: Third-year pharmacy students (N=13) completed the Attitudes toward Psychiatry-30 (ATP-30) questionnaire before and after taking an active learning, advanced psychopharmacology course. RESULTS: ATP-30 questionnaires indicated that the course resulted in a positive change in students' attitudes toward psychiatric patients and treatment. CONCLUSION: Considering the increasing role of rural pharmacists in the care of patients, it is suggested that colleges of pharmacy that emphasize rural practice should augment mental health related education. PMID- 19190300 TI - A novel approach to medicine training for psychiatry residents. AB - OBJECTIVE: A unique rotation was developed to address limited outpatient internal medicine training in psychiatric residency by the University of California, Davis, Department of Psychiatry and Behavioral Sciences, which provides medical care to patients with mental illness. METHODS: The number of patients seen by the service and the number of psychiatric consults was determined from electronic records for the 2005-2006 academic year. Evaluations by psychiatry residents completing the rotation were reviewed. Three internist-psychiatrists and one family medicine-psychiatrist provided supervision. RESULTS: A total of 1,255 patients were treated during the 2005-2006 academic year. The quality of the educational experience was positive, with an overall rating of 4.43 on a scale from 1 to 5, with 5 being the highest. CONCLUSION: Training psychiatry residents in internal medicine can be better integrated into their psychiatry education in a creative fashion when the teaching and supervision is provided by jointly trained attendings in internal medicine/psychiatry or family medicine/psychiatry. The success of the rotation contributed to the development of a combined internal medicine and psychiatry residency program. PMID- 19190301 TI - Supervising the supervisors: a case-based group forum for faculty development. AB - OBJECTIVE: Faculty development has increasingly become a focus in medical education. Many models are employed around the United States. The authors present a group model process developed to train new educators to teach our medical students. METHODS: The authors met monthly with a diverse faculty over the past 4 years and interacted in a nonhierarchal manner. The authors reviewed the literature, supervised one another, and developed a participatory method of solving educational and administrative problems. RESULTS: What emerged was a "case-based," collaborative group format to develop an educational philosophy, address challenging supervisory problems, develop tangible educational materials and "products," and promote academic careers. CONCLUSION: The authors propose this model for faculty development. PMID- 19190303 TI - University of Colorado department of psychiatry evidence-based medicine educational project. PMID- 19190304 TI - A 76-year-old man with recurrent Clostridium difficile-associated diarrhea: review of C. difficile infection. AB - Clostridium difficile infection (CDI) is a common and increasingly severe nosocomial infectious disease. The case of Mr S, a 76-year-old man with multiple recurrences of CDI, illustrates the difficulties in treating recurrent disease and the way it complicates the management of other medical conditions. Risk factors for CDI include antimicrobial use, hospital admission, advancing age, and severe underlying disease. A clinical diagnosis of CDI is usually confirmed by identifying C. difficile toxins in a stool sample. Evidence supports metronidazole, 500 mg every 6 hours for 10 to 14 days, as the treatment of choice for mild to moderately severe CDI. Oral vancomycin, 125 mg every 6 hours for 10 to 14 days, is recommended for severe CDI, for which it is more effective than metronidazole. Recurrent CDI occurs in more than 20% of patients when metronidazole or vancomycin treatment is discontinued. Few studies have evaluated treatment options for recurrent CDI, but a prolonged, tapering, and pulse-dosed regimen of oral vancomycin is commonly used. Careful attention to antimicrobial stewardship and infection control practices is essential to curb this nosocomial, iatrogenic disease. PMID- 19190305 TI - The cover. Black Indian-Buffalo Soldiers. PMID- 19190306 TI - A piece of my mind. The first wake. PMID- 19190307 TI - Melamine scandals highlight hazards of increasingly globalized food chain. PMID- 19190308 TI - Guideline provides evidence-based advice for treating osteoarthritis of the knee. PMID- 19190309 TI - Researchers seek MRI-safe pacemakers. PMID- 19190310 TI - Evidence-based evaluation of physician work hour regulations. PMID- 19190311 TI - Reporting system for violent deaths and youth suicide. PMID- 19190312 TI - A view on the room. PMID- 19190313 TI - Mechanical reperfusion and long-term mortality in patients with acute myocardial infarction presenting 12 to 48 hours from onset of symptoms. PMID- 19190314 TI - Estimated radiation dose associated with cardiac CT angiography. AB - CONTEXT: Cardiac computed tomography (CT) angiography (CCTA) has emerged as a useful diagnostic imaging modality in the assessment of coronary artery disease. However, the potential risks due to exposure to ionizing radiation associated with CCTA have raised concerns. OBJECTIVES: To estimate the radiation dose of CCTA in routine clinical practice as well as the association of currently available strategies with dose reduction and to identify the independent factors contributing to radiation dose. DESIGN, SETTING, AND PATIENTS: A cross-sectional, international, multicenter, observational study (50 study sites: 21 university hospitals and 29 community hospitals) of estimated radiation dose in 1965 patients undergoing CCTA between February and December 2007. Linear regression analysis was used to identify independent predictors associated with dose. MAIN OUTCOME MEASURE: Dose-length product (DLP) of CCTA. RESULTS: The median DLP of 1965 CCTA examinations performed at 50 study sites was 885 mGy x cm (interquartile range, 568-1259 mGy x cm), which corresponds to an estimated radiation dose of 12 mSv (or 1.2 x the dose of an abdominal CT study or 600 chest x-rays). A high variability in DLP was observed between study sites (range of median DLPs per site, 331-2146 mGy x cm). Independent factors associated with radiation dose were patient weight (relative effect on DLP, 5%; 95% confidence interval [CI], 4%-6%), absence of stable sinus rhythm (10%; 95% CI, 2%-19%), scan length (5%; 95% CI, 4%-6%), electrocardiographically controlled tube current modulation (-25%; 95% CI, -23% to -28%; applied in 73% of patients), 100-kV tube voltage (-46%; 95% CI, -42% to -51%; applied in 5% of patients), sequential scanning (-78%; 95% CI, -77% to -79%; applied in 6% of patients), experience in cardiac CT (-1%; 95% CI, -1% to 0%), number of CCTAs per month (0%; 95% CI, 0% 1%), and type of 64-slice CT system (for highest vs lowest dose system, 97%; 95% CI, 88%-106%). Algorithms for dose reduction were not associated with deteriorated diagnostic image quality in this observational study. CONCLUSIONS: Median doses of CCTA differ significantly between study sites and CT systems. Effective strategies to reduce radiation dose are available but some strategies are not frequently used. The comparable diagnostic image quality may support an increased use of dose-saving strategies in adequately selected patients. PMID- 19190315 TI - Altitude and all-cause mortality in incident dialysis patients. AB - CONTEXT: Patients undergoing dialysis at higher altitude receive lower erythropoietin doses, yet achieve higher hemoglobin concentrations. Increased iron availability caused by activation of hypoxia-induced factors at higher altitude may explain this finding. Hypoxia-induced factors are also involved in other pathways that may affect morbidity and mortality. OBJECTIVE: To study whether mortality differed by altitude in patients initiating dialysis. DESIGN, SETTING, AND PARTICIPANTS: Retrospective cohort of patients initiating dialysis in the United States between 1995 and 2004. Patients were stratified by the average elevation of their residential zip code. Covariates included age, sex, race, Medicaid coverage, dialysis modality, comorbidities, and reported laboratory measurements. We constructed proportional hazards models of all-cause mortality, stratifying by year, and censoring patients at 5 years from first dialysis, at the end of the database (December 31, 2004), or loss to follow-up. We also compared age- and sex-adjusted standardized mortality rates of US patients receiving dialysis with the general population. MAIN OUTCOME MEASURE: Mortality from any cause. RESULTS: A total of 804 812 patients initiated dialysis and were followed up for a median of 1.78 years. Crude mortality rates per 1000 person-years were 220.1 at an altitude lower than 76 m (<250 ft), 221.2 from 76 through 609 m (250-1999 ft), 214.6 from 610 through 1218 m (2000-3999 ft), 184.9 from 1219 through 1828 m (4000 to 5999 ft), and 177.2 at an altitude higher than 1828 m (>6000 ft). After multivariable adjustment, compared with patients living at an altitude of lower than 76 m, the relative mortality rates were 0.97 (95% confidence interval [CI], 0.96-0.98) for those living from 76 through 609 m; 0.93 (95% CI, 0.91-0.95), from 610 through 1218 m; 0.88 (95% CI, 0.84-0.91), from 1219 through 1828 m, and 0.85 (95% CI, 0.79-0.92) higher than 1828 m. Age- and sex standardized mortality decreased more with altitude in patients receiving dialysis than in the general population. CONCLUSIONS: Altitude was inversely associated with all-cause mortality among US patients receiving dialysis. PMID- 19190316 TI - Mortality risk associated with low-trauma osteoporotic fracture and subsequent fracture in men and women. AB - CONTEXT: There are few data on long-term mortality following osteoporotic fracture and fewer following subsequent fracture. OBJECTIVES: To examine long term mortality risk in women and men following all osteoporotic fractures and to assess the association of subsequent fracture with that risk. DESIGN, SETTING, AND PARTICIPANTS: Prospective cohort from the Dubbo Osteoporosis Epidemiology Study of community-dwelling women and men aged 60 years and older from Dubbo, Australia, who sustained a fracture between April 1989 and May 2007. MAIN OUTCOME MEASURES: Age- and sex-specific standardized mortality ratios (SMRs) compared with the overall Dubbo population for hip, vertebral, major, and minor fractures. RESULTS: In women, there were 952 low-trauma fractures followed by 461 deaths, and in men, 343 fractures were followed by 197 deaths. Age-adjusted SMRs were increased following hip fractures (SMRs, 2.43 [95% confidence interval [CI], 2.02 2.93] and 3.51 [95% CI, 2.65-4.66]), vertebral fractures (SMRs, 1.82 [95% CI, 1.52-2.17] and 2.12 [95% CI, 1.66-2.72]), major fractures (SMRs, 1.65 [95% CI, 1.31-2.08] and 1.70 [95% CI, 1.23-2.36]), and minor fractures (SMRs, 1.42 [95% CI, 1.19-1.70] and 1.33 [95% CI, 0.99-1.80]) for both women and men, respectively. Mortality was increased for all ages for all fractures except minor fractures for which increased mortality was only apparent for those older than 75 years. Increased mortality risk persisted for 5 years for all fractures and up to 10 years for hip fractures. Increases in absolute mortality that were above expected, for 5 years after fracture, ranged from 1.3 to 13.2 per 100 person years in women and from 2.7 to 22.3 per 100 person-years in men, depending on fracture type. Subsequent fracture was associated with an increased mortality hazard ratio of 1.91 (95% CI, 1.54-2.37) in women and 2.99 (95% CI, 2.11-4.24) in men. Mortality risk following a subsequent fracture then declined but beyond 5 years still remained higher than in the general population (SMR, 1.41 [95% CI, 1.01-1.97] and SMR, 1.78 [95% CI, 0.96-3.31] for women and men, respectively). Predictors of mortality after any fragility fracture for both men and women included age, quadriceps weakness, and subsequent fracture but not comorbidities. Low bone mineral density, having smoked, and sway were also predictors for women and less physical activity for men. CONCLUSIONS: In a sample of older women and men, all low-trauma fractures were associated with increased mortality risk for 5 to 10 years. Subsequent fracture was associated with increased mortality risk for an additional 5 years. PMID- 19190317 TI - Update: A 27-year-old woman with a diagnosis of polycystic ovary syndrome. PMID- 19190318 TI - Obesity and the economy: from crisis to opportunity. PMID- 19190319 TI - A closer look at the economic argument for disease prevention. PMID- 19190320 TI - Smoking as a factor in causing lung cancer. PMID- 19190321 TI - Radiation protection of patients undergoing cardiac computed tomographic angiography. PMID- 19190322 TI - JAMA patient page. Bipolar disorder. PMID- 19190323 TI - Galectin-3 mediates nuclear beta-catenin accumulation and Wnt signaling in human colon cancer cells by regulation of glycogen synthase kinase-3beta activity. AB - Wnt/beta-catenin signaling plays an essential role in colon carcinogenesis. Galectin-3, a beta-galactoside-binding protein, has been implicated in Wnt signaling, but the precise mechanisms by which galectin-3 modulates the Wnt pathway are unknown. In the present study, we determined the effects of galectin 3 on the Wnt/beta-catenin pathway in colon cancer cells, as well as the mechanisms involved. Galectin-3 levels were manipulated in human colon cancer cells by stable transfection of galectin-3 antisense, short hairpin RNA, or full length galectin-3 cDNA, and effects on beta-catenin levels, subcellular distribution, and Wnt signaling were determined. Galectin-3 levels correlated with beta-catenin levels in a variety of colon cancer cell lines. Down-regulation of galectin-3 resulted in decreased beta-catenin protein levels but no change in beta-catenin mRNA levels, suggesting that galectin-3 modulates beta-catenin by another mechanism. Reduction of galectin-3 led to reduced nuclear beta-catenin with a concomitant decrease in TCF4 transcriptional activity and expression of its target genes. Conversely, transfection of galectin-3 cDNA into colon cancer cells increased beta-catenin expression and TCF4 transcriptional activity. Down regulation of galectin-3 resulted in AKT and glycogen synthase kinase-3beta (GSK 3beta) dephosphorylation and increased GSK activity, increasing beta-catenin phosphorylation and degradation. Ly294002, an inhibitor of phosphatidylinositol 3 kinase, and dominant-negative AKT, suppressed TCF4 transcriptional activity induced by galectin-3 whereas LiCl, a GSK-3beta inhibitor, increased TCF4 activity, mimicking the effects of galectin-3. These results suggest that galectin-3 mediates Wnt signaling, at least in part, by regulating GSK-3beta phosphorylation and activity via the phosphatidylinositol 3-kinase/AKT pathway, and, thus, the degradation of beta-catenin in colon cancer cells. PMID- 19190324 TI - Inhibition of eIF2alpha dephosphorylation maximizes bortezomib efficiency and eliminates quiescent multiple myeloma cells surviving proteasome inhibitor therapy. AB - The proteasome inhibitor bortezomib (Velcade) effectively eradicates multiple myeloma (MM) cells, partly by activating endoplasmic reticulum (ER) stress apoptotic signaling. However, MM recurrences in bortezomib-treated patients are invariable. We have shown that ER stress signaling can also induce growth arrest and survival in cancer cells. Thus, we hypothesized that bortezomib therapy could induce quiescence and survival of residual MM cells, contributing to disease recurrence. Here, we report that in MM cells, proteasome inhibition with MG-132 or bortezomib results in a surviving cell fraction that enters a prolonged quiescent state (G(0)-G(1) arrest). Mechanism analysis revealed that bortezomib surviving quiescent cells attenuate eIF2alpha phosphorylation and induction of the ER stress proapoptotic gene GADD153. This occurs independently of the eIF2alpha upstream kinases PERK, GCN2, and PKR. In contrast, the prosurvival ER chaperone BiP/Grp78 was persistently induced. The bortezomib-surviving quiescent fraction could be eradicated by a simultaneous or sequential combination therapy with salubrinal, an inhibitor of GADD34-PP1C phosphatase complex, and, in consequence, eIF2alpha dephosphorylation. This effect was mimicked by expression of a phosphorylated mimetic eIF2alpha-S51D mutant. Our data indicate that bortezomib can induce growth arrest in therapy-surviving MM cells and that attenuation of eIF2alpha phosphorylation contributes to this survival. Most importantly, this survival mechanism can be blocked by inhibiting eIF2alpha dephosphorylation. Thus, strategies that maintain eIF2alpha in a hyperphosphorylated state may be a novel therapeutic approach to maximize bortezomib-induced apoptosis and reduce residual disease and recurrences in this type of cancer. PMID- 19190325 TI - Functional analysis of 11q13.5 amplicon identifies Rsf-1 (HBXAP) as a gene involved in paclitaxel resistance in ovarian cancer. AB - The chromosome 11q13.5 locus is frequently amplified in several types of human cancer. We have previously shown that 11q13.5 amplification was associated with significantly shorter overall survival in ovarian cancer patients, but the molecular mechanisms of how amplification of this locus contributes to disease aggressiveness remain unclear. Because ovarian cancer mortality is primarily related to resistance of chemotherapeutic agents, we screened the top six candidate genes within this amplicon for their contribution to drug resistance. Rsf-1 (also known as HBXAP) was found to be the only gene in which gene knockdown sensitized tumor cells to paclitaxel. Rsf-1 has been known to interact with hSNF2H to form an ISWI chromatin remodeling complex. We found that Rsf-1 was up regulated in paclitaxel-resistant ovarian cancer cell lines, and Rsf-1 immunoreactivity in primary ovarian carcinoma tissues correlated with in vitro paclitaxel resistance. Ectopic expression of Rsf-1 significantly enhanced paclitaxel resistance in ovarian cancer cells. Down-regulation of hSNF2H or disruption of hSNF2H and Rsf-1 interaction enhanced paclitaxel sensitivity in tumor cells with Rsf-1 up-regulation. Rsf-1 expression altered expression in several genes and activated certain signaling pathways that may contribute to drug resistance. In conclusion, our results suggest that Rsf-1 is the major gene within the 11q13.5 amplicon that contributes to paclitaxel resistance, and the formation of the Rsf-1/hSNF2H complex is required for inducing this phenotype. PMID- 19190326 TI - Breast cancer metastasis suppressor 1 up-regulates miR-146, which suppresses breast cancer metastasis. AB - Breast cancer metastasis suppressor 1 (BRMS1) is a predominantly nuclear protein that differentially regulates expression of multiple genes, leading to suppression of metastasis without blocking orthotopic tumor growth in multiple human and murine cancer cells of diverse origins. We hypothesized that miR-146 may be involved in the ability of BRMS1 to supress metastasis because miR-146 expression is altered by BRMS1 and because BRMS1 and miR-146 are both associated with decreased signaling through the nuclear factor-kappaB pathway. BRMS1 significantly up-regulates miR-146a by 6- to 60-fold in metastatic MDA-MB-231 and MDA-MB-435 cells, respectively, and miR-146b by 40-fold in MDA-MB-435 as measured by real-time quantitative reverse transcription-PCR. Transduction of miR-146a or miR-146b into MDA-MB-231 down-regulated expression of epidermal growth factor receptor, inhibited invasion and migration in vitro, and suppressed experimental lung metastasis by 69% and 84%, respectively (mean +/- SE: empty vector = 39 +/- 6, miR-146a = 12 +/- 1, miR-146b = 6 +/- 1). These results further support the recent notion that modulating the levels of miR-146a or miR-146b could have a therapeutic potential to suppress breast cancer metastasis. PMID- 19190327 TI - Zerumbone abolishes RANKL-induced NF-kappaB activation, inhibits osteoclastogenesis, and suppresses human breast cancer-induced bone loss in athymic nude mice. AB - Receptor activator of nuclear factor-kappaB (NF-kappaB) ligand (RANKL) has emerged as a major mediator of bone resorption, commonly associated with cancer and other chronic inflammatory diseases. Inhibitors of RANKL signaling thus have potential in preventing bone loss. In the present report, the potential of zerumbone, a sesquiterpene derived from subtropical ginger, to modulate osteoclastogenesis induced by RANKL and breast cancer was examined. We found that zerumbone inhibited RANKL-induced NF-kappaB activation in mouse monocyte, an osteoclast precursor cell, through inhibition of activation of IkappaBalpha kinase, IkappaBalpha phosphorylation, and IkappaBalpha degradation. Zerumbone also suppressed RANKL-induced differentiation of these cells to osteoclasts. This sesquiterpene also inhibited the osteoclast formation induced by human breast tumor cells and by multiple myeloma cells. Finally, we examined whether zerumbone could prevent human breast cancer-induced bone loss in animals. We found that zerumbone decreased osteolysis in a dose-dependent manner in MDA-MB-231 breast cancer tumor-bearing athymic nude mice. These results indicate that zerumbone is an effective blocker of RANKL-induced NF-kappaB activation and of osteoclastogenesis induced by RANKL and tumor cells, suggesting its potential as a therapeutic agent for osteoporosis and cancer-associated bone loss. PMID- 19190328 TI - In vivo off-resonance saturation magnetic resonance imaging of alphavbeta3 targeted superparamagnetic nanoparticles. AB - Magnetic resonance imaging is a powerful clinical imaging technique that allows for noninvasive tomographic visualization of anatomic structures with high spatial resolution and soft tissue contrast. However, its application in molecular imaging of cancer has been limited by the lack of sensitivity and detection accuracy in depicting the biochemical expression of these diseases. Here, we combine an ultrasensitive design of superparamagnetic polymeric micelles (SPPM) and an off-resonance saturation (ORS) method to enhance the imaging efficacy of tumor biomarkers in vivo. SPPM nanoparticles encoded with cyclic(RGDfK) were able to target the alpha(v)beta(3)-expressing microvasculature in A549 non-small cell lung tumor xenografts in mice. ORS greatly improved tumor detection accuracy over the conventional T(2)*-weighted method by its ability to turn "ON" the contrast of SPPM. This combination of ORS imaging with a tumor vasculature-targeted, ultrasensitive SPPM design offers new opportunities in molecular imaging of cancer. PMID- 19190329 TI - Whole genome comparison of allelic imbalance between noninvasive and invasive small-sized lung adenocarcinomas. AB - Seventy-two small-sized (IA) and associated with worse prognoses (P = 0.04) and, thus, would be involved in invasion and/or metastasis of adenocarcinoma cells and useful for the prediction of prognosis of patients with small-sized lung adenocarcinoma. PMID- 19190330 TI - Human renal cancer cells express a novel membrane-bound interleukin-15 that induces, in response to the soluble interleukin-15 receptor alpha chain, epithelial-to-mesenchymal transition. AB - Although interleukin-15 (IL-15) is a powerful immunomodulatory factor that has been proposed for cancer immunotherapy, its intratumoral expression may be correlated with tumor progression and/or poor clinical outcome. Therefore, neoplasias potentially sensitive to immunotherapy should be checked for their IL 15 expression and function before choosing immunotherapy protocols. Primary human renal cancer cells (RCC) express a novel form of membrane-bound IL-15 (mb-IL-15), which displays three major original properties: (a) It is expressed as a functional membrane homodimer of 27 kDa, (b) it is shed in the extracellular environment by the metalloproteases ADAM17 and ADAM10, and (c) its stimulation by soluble IL-15 receptor alpha (s-IL-15Ralpha) chain triggers a complex reverse signal (mitogen-activated protein kinases, FAK, pMLC) necessary and sufficient to ~induce epithelial-mesenchymal transdifferentiation (EMT), a crucial process in tumor progression whose induction is unprecedented for IL-15. In these cells, complete EMT is characterized by a dynamic reorganization of the cytoskeleton with the subsequent generation of a mesenchymal/contractile phenotype (alpha-SMA and vimentin networks) and the loss of the epithelial markers E-cadherin and ZO 1. The retrosignaling functions are, however, hindered through an unprecedented cytokine/receptor interaction of mb-IL-15 with membrane-associated IL-15Ralpha subunit that tunes its signaling potential competing with low concentrations of the s-IL-15Ralpha chain. Thus, human RCC express an IL-15/IL-15R system, which displays unique biochemical and functional properties that seem to be directly involved in renal tumoral progression. PMID- 19190331 TI - A high throughput, whole cell screen for small molecule inhibitors of the mitotic spindle checkpoint identifies OM137, a novel Aurora kinase inhibitor. AB - In mitosis, the kinetochores of chromosomes that lack full microtubule attachments and/or mechanical tension activate a signaling pathway called the mitotic spindle checkpoint that blocks progression into anaphase and prevents premature segregation of the chromatids until chromosomes become aligned at the metaphase plate. The spindle checkpoint is responsible for arresting cells in mitosis in response to chemotherapeutic spindle poisons such as paclitaxel or vinblastine. Some cancer cells show a weakened checkpoint signaling system that may contribute to chromosome instability in tumors. Because complete absence of the spindle checkpoint leads to catastrophic cell division, we reasoned that drugs targeting the checkpoint might provide a therapeutic window in which the checkpoint would be eliminated in cancer cells but sufficiently preserved in normal cells. We developed an assay to identify lead compounds that inhibit the spindle checkpoint. Most cells respond to microtubule drugs by activating the spindle checkpoint and arresting in mitosis with a rounded morphology. Our assay depended on the ability of checkpoint inhibitor compounds to drive mitotic exit and cause cells to flatten onto the substrate in the continuous presence of microtubule drugs. In this study, we characterize one of the compounds, OM137, as an inhibitor of Aurora kinases. We find that this compound is growth inhibitory to cultured cells when applied at high concentration and potentiates the growth inhibitory effects of subnanomolar concentrations of paclitaxel. PMID- 19190332 TI - An activating beta1 integrin mutation increases the conversion of benign to malignant skin tumors. AB - Identifying the physiologic relevance of cancer-associated genetic polymorphisms is a major challenge. Several changes in the coding sequence of beta integrin subunits have now been described in human tumors. One of these, T188Ibeta1, was identified as a heterozygous mutation in a poorly differentiated squamous cell carcinoma (SCC) and shown to activate extracellular matrix adhesion and inhibit keratinocyte differentiation in vitro. To study its contribution to tumor development, we overexpressed the mutant or wild-type (WT) human beta1 subunit in the basal layer of mouse epidermis using the keratin 14 promoter. The transgenic integrins were expressed at the cell surface and were functional, with the T188Ibeta1 subunit promoting cell spreading to a greater extent than WTbeta1. Epidermal proliferation and differentiation were unaffected and no expansion of the stem cell compartment was detected. During chemical carcinogenesis, both transgenes increased papilloma formation, but only the T188Ibeta1 transgene stimulated the conversion of papillomas to SCCs. Papillomas bearing the mutation showed increased Erk activity and reduced differentiation. SCCs expressing T188Ibeta1 were less well-differentiated than those expressing WTbeta1. These observations establish that the expression of a genetic variant in the I-like domain of beta1 integrins does not affect normal epidermal homeostasis, but increases tumor susceptibility and influences tumor type. PMID- 19190333 TI - Telomere attrition in cancer cells and telomere length in tumor stroma cells predict chromosome instability in esophageal squamous cell carcinoma: a genome wide analysis. AB - Previous studies showed that chromosomal instability was common in esophageal squamous cell carcinoma (ESCC); however, the mechanisms underlying this instability are unknown. Individuals with deficiencies in telomere maintenance are susceptible to enhanced telomere loss during cell proliferation; such deficiencies could result in telomere dysfunction and genomic instability. We investigated the association between genome-wide chromosomal changes in cancer cells and telomere length/attrition in cancer/stroma cells in 47 ESCC patients. Genome-wide detection of loss of heterozygosity was performed using the Affymetrix GeneChip single nucleotide polymorphism arrays. Telomere length was assessed separately for cancer cells, carcinoma-associated fibroblasts (CAF), infiltrative lymphocytes, and adjacent normal epithelial cells by quantitative fluorescent in situ hybridization using paraffin-embedded sections. Telomere length differed significantly among cell types, such that length in infiltrative lymphocytes > CAFs > cancer cells. Shortened telomeres were observed in cancer cells in 44 of 47 (94%) of the tumors examined. Telomere length in CAFs was significantly associated with chromosomal instability on 4q and 13q and lymphocyte telomere length was significantly associated with instability on chromosomal arms 15q. Although telomere length in cancer cells was not associated with chromosome arm instability, telomere attrition in cancer cells, defined as the telomere length in CAFs minus the telomere length in cancer cells, was significantly associated with chromosomal instability on 13q and 15q. This study provides evidence that telomere shortening is a common genetic alteration in ESCC and that chromosome arm instability is related to both telomere attrition in cancer cells and telomere length in tumor stroma cells. PMID- 19190334 TI - Altered proliferation and differentiation properties of primary mammary epithelial cells from BRCA1 mutation carriers. AB - Female BRCA1 mutation carriers have a nearly 80% probability of developing breast cancer during their life-time. We hypothesized that the breast epithelium at risk in BRCA1 mutation carriers harbors mammary epithelial cells (MEC) with altered proliferation and differentiation properties. Using a three-dimensional culture technique to grow MECs ex vivo, we found that the ability to form colonies, an indication of clonality, was restricted to the aldehyde dehydrogenase 1-positive fraction in MECs but not in HCC1937 BRCA1-mutant cancer cells. Primary MECs from BRCA1 mutation carriers (n = 9) had a 28% greater ability for clonal growth compared with normal controls (n = 6; P = 0.006), and their colonies were significantly larger. Colonies in controls and BRCA1 mutation carriers stained positive for BRCA1 by immunohistochemistry, and 79% of the examined single colonies from BRCA1 carriers retained heterozygosity for BRCA1 (ROH). Colonies from BRCA1 mutation carriers frequently showed high epidermal growth factor receptor (EGFR) expression (71% EGFR positive versus 44% in controls) and were negative for estrogen receptor (ERalpha; 32% ER negative, 44% mixed, 24% ER positive versus 90% ER positive in controls). Expression of CK14 and p63 were not significantly different. Microarray studies revealed that colonies from BRCA1 mutant PMECs anticipate expression profiles found in BRCA1-related tumors, and that the EGFR pathway is up-regulated. We conclude that BRCA1 haploinsufficiency leads to an increased ability for clonal growth and proliferation in the PMECs of BRCA1 mutation carriers, possibly as a result of EGFR pathway activation. These altered growth and differentiation properties may render BRCA1-mutant PMECs vulnerable to transformation and predispose to the development of ER-negative, EGFR-positive breast cancers. PMID- 19190335 TI - Effective immunotherapy against murine gliomas using type 1 polarizing dendritic cells--significant roles of CXCL10. AB - In an attempt to develop effective vaccines against central nervous system (CNS) tumors, we evaluated the ability of vaccines with standard dendritic cells (DC) versus type 1 polarizing DCs (DC1) to induce glioma-specific type 1 CTLs with CNS tumor-relevant homing properties and the mechanism of their action. C57BL/6 mouse derived bone marrow cells were cultured with mouse granulocyte/macrophage colony stimulating factor (GM-CSF) for 6 days, and CD11c(+) cells were subsequently cultured with GM-CSF, rmIFN-gamma, rmIFN-alpha, rmIL-4, and polyinosinic polycytidylic acid stabilized by lysine and carboxymethylcellulose for 24 hours to generate DC1s. In analogy to their human counterparts, mouse DC1s exhibited surface marker profiles of mature DCs and produced high levels of IL-12 and CXCL10. Importantly for their application as cancer vaccines, such DC1s stably retained their type 1 phenotype even when exposed to type 2-promoting or regulatory T cell (Treg)-promoting environments. Consistently, mouse DC1s induced antigen-specific type 1 CTLs more efficiently than nonpolarized DCs in vitro. DC1s given s.c. migrated into draining lymph nodes, induced antigen-specific CTLs, and suppressed Treg accumulation. In addition, s.c. immunization with DC1s loaded with glioma-associated antigen (GAA)-derived CTL epitope peptides prolonged the survival of CNS GL261 glioma-bearing mice, which was associated with efficient CNS glioma homing of antigen-specific CTLs. Intratumoral injections of GAA peptide-loaded DC1s further enhanced the anti-CNS glioma effects of DC1-based s.c. immunization. Interestingly, the antitumor functions were abrogated with CXCL10(-/-) mouse-derived DC1s. Collectively, these findings show the anti-CNS glioma effects of DC1-based therapy and a novel role of CXCL10 in the immunologic and therapeutic activity of DC-based cancer vaccines. PMID- 19190336 TI - Identification of a novel pathway that selectively modulates apoptosis of breast cancer cells. AB - Expression of the nuclear receptor interacting factor 3 (NRIF3) coregulator in a wide variety of breast cancer cells selectively leads to rapid caspase-2 dependent apoptotic cell death. A novel death domain (DD1) was mapped to a 30 amino acid region of NRIF3. Because the cytotoxicity of NRIF3 and DD1 seems to be cell type-specific, these studies suggest that breast cancer cells contain a novel "death switch" that can be specifically modulated by NRIF3 or DD1. Using an MCF-7 cell cDNA library in a yeast two-hybrid screen, we cloned a factor that mediates apoptosis by DD1 and refer to this factor as DD1-interacting factor-1 (DIF-1). DIF-1 is a transcriptional repressor that mediates its effect through SirT1, and this repression is attenuated by the binding of NRIF3/DD1. DIF-1 expression rescues breast cancer cells from NRIF3/DD1-induced apoptosis. Small interfering RNA (siRNA) knockdown of DIF-1 selectively leads to apoptosis of breast cancer cells, further suggesting that DIF-1 plays a key role in NRIF3/DD1 mediated apoptosis. A protein kinase A inhibitor (H89) also elicits apoptosis of breast cancer cells but not of the other cell types examined, and DIF-1 also protects these cells from H89-mediated apoptosis. In addition, H89 incubation results in a rapid increase in NRIF3 levels and siRNA knockdown of NRIF3 protects breast cancer cells from H89-mediated apoptosis. Our results indicate that DIF-1 plays a key role in breast cancer cell survival and further characterizing this pathway may provide important insights into developing novel therapies to selectively target breast cancer cells for apoptosis. PMID- 19190337 TI - Suppressor of cytokine signaling 3 promotes bone marrow cells to differentiate into CD8+ T lymphocytes in lung tissue via up-regulating Notch1 expression. AB - Suppressor of cytokine signaling 3 (SOCS3) expression in bone marrow cells (BMC) was up-regulated upon exposure to interleukin 6, lipopolysaccharide, or tumor associated factors. But, how the up-regulated SOCS3 affects differentiation of BMCs is incompletely characterized. Here, we showed that SOCS3 promoted BMCs to intently differentiate into CD8 T cells. Importantly, lung can be as one athymus tissue for the BMCs to differentiate into CD8(+) T cells. Notch1 plays a critical role in the differentiation from SOCS3-transfected BMCs to CD8(+) T cells. We conclude that the up-regulated SOCS3 in some pathologic conditions, such as tumor and inflammation, might promote BMCs to differentiate into CD8(+) T lymphocytes in lung tissue via up-regulating Notch1 expression. This may represent a new mechanism against diseases such as tumor. PMID- 19190338 TI - Identification of SULF2 as a novel transcriptional target of p53 by use of integrated genomic analyses. AB - Microarray analysis has been useful for identifying the targets of many transcription factors. However, gene expression changes in response to transcription factor perturbation reveal both direct transcriptional targets and secondary gene regulation. By integrating RNA interference, gene expression profiling, and chromatin immunoprecipitation technologies, we identified a set of 32 direct transcriptional targets of the tumor suppressor p53. Of these 32 genes, 11 are not currently associated with the core p53 pathway. From among these novel pathway members, we focused on understanding the connection between p53 and SULF2, which encodes an extracellular heparan sulfate 6-O-endosulfatase that modulates the binding of growth factors to their cognate receptors and that has been shown to function as a tumor suppressor. Genetic and pharmacologic perturbation of p53 directly influences SULF2 expression, and similar to silencing of TP53, RNA interference-mediated suppression of SULF2 results in an impaired senescence response of cells to genotoxic stress. Thus, our integrated genomic approach has led to the identification of a novel mediator of p53 network biology. PMID- 19190339 TI - Breast cancer cell lines contain functional cancer stem cells with metastatic capacity and a distinct molecular signature. AB - Tumors may be initiated and maintained by a cellular subcomponent that displays stem cell properties. We have used the expression of aldehyde dehydrogenase as assessed by the ALDEFLUOR assay to isolate and characterize cancer stem cell (CSC) populations in 33 cell lines derived from normal and malignant mammary tissue. Twenty-three of the 33 cell lines contained an ALDEFLUOR-positive population that displayed stem cell properties in vitro and in NOD/SCID xenografts. Gene expression profiling identified a 413-gene CSC profile that included genes known to play a role in stem cell function, as well as genes such as CXCR1/IL-8RA not previously known to play such a role. Recombinant interleukin 8 (IL-8) increased mammosphere formation and the ALDEFLUOR-positive population in breast cancer cell lines. Finally, we show that ALDEFLUOR-positive cells are responsible for mediating metastasis. These studies confirm the hierarchical organization of immortalized cell lines, establish techniques that can facilitate the characterization of regulatory pathways of CSCs, and identify potential stem cell markers and therapeutic targets. PMID- 19190340 TI - Wilms' tumor 1 gene expression in hepatocellular carcinoma promotes cell dedifferentiation and resistance to chemotherapy. AB - The Wilms' tumor 1 gene (WT1) encodes a transcription factor involved in cell growth and development. As we previously reported, WT1 expression is hardly detectable in normal hepatic tissue but is induced in liver cirrhosis. Although WT1 has been found to be overexpressed in a number of malignancies, the role of WT1 in hepatocarcinogenesis has not been clarified. We found that WT1 is expressed in several human hepatocellular carcinoma (HCC) cell lines, including PLC/PRF/5 and HepG2, and in HCC tumor tissue in 42% of patients. WT1 small interfering RNAs did not affect proliferation rate of HCC cells but abrogated their resistance to anoikis. Transcriptome analysis of PLC/PRF/5 cells after WT1 knockdown showed up-regulation of 251 genes and down-regulation of 321. Ninety percent of the former corresponded to metabolic genes, mostly those characterizing the mature hepatocyte phenotype. On the contrary, genes that decreased upon WT1 inhibition were mainly related to defense against apoptosis, cell cycle, and tumor progression. In agreement with these findings, WT1 expression increased the resistance of liver tumor cells to doxorubicin, a compound used to treat HCC. Interestingly, doxorubicin strongly enhanced WT1 expression in both HCC cells and normal human hepatocytes. Among different chemotherapeutics, induction of WT1 transcription was restricted to topoisomerase 2 inhibitors. When WT1 expression was prohibited, doxorubicin caused a marked increase in caspase-3 activation. In conclusion, WT1 is expressed in a substantial proportion of HCC contributing to tumor progression and resistance to chemotherapy, suggesting that WT1 may be an important target for HCC treatment. PMID- 19190341 TI - Correlation analysis between single-nucleotide polymorphism and expression arrays in gliomas identifies potentially relevant target genes. AB - Primary brain tumors are a major cause of cancer mortality in the United States. Therapy for gliomas, the most common type of primary brain tumors, remains suboptimal. The development of improved therapeutics will require greater knowledge of the biology of gliomas at both the genomic and transcriptional levels. We have previously reported whole genome profiling of chromosome copy number alterations (CNA) in gliomas, and now present our findings on how those changes may affect transcription of genes that may be involved in tumor induction and progression. By calculating correlation values of mRNA expression versus DNA copy number average in a moving window around a given RNA probe set, biologically relevant information can be gained that is obscured by the analysis of a single data type. Correlation coefficients ranged from -0.6 to 0.7, highly significant when compared with previous studies. Most correlated genes are located on chromosomes 1, 7, 9, 10, 13, 14, 19, 20, and 22, chromosomes known to have genomic alterations in gliomas. Additionally, we were able to identify CNAs whose gene expression correlation suggests possible epigenetic regulation. This analysis revealed a number of interesting candidates such as CXCL12, PTER, and LRRN6C, among others. The results have been verified using real-time PCR and methylation sequencing assays. These data will further help differentiate genes involved in the induction and/or maintenance of the tumorigenic process from those that are mere passenger mutations, thereby enriching for a population of potentially new therapeutic molecular targets. PMID- 19190342 TI - Expression of human organic cation transporter 3 in kidney carcinoma cell lines increases chemosensitivity to melphalan, irinotecan, and vincristine. AB - Renal cell carcinoma (RCC) is usually chemoresistant. This chemoresistance could be overcome if specific cytostatics are applied for which the RCC expresses an uptake transporter. In the present study, we investigated the expression of solute carrier (SLC) transporters in different RCC lines and their ability to interact with chemotherapeutics. We tested five RCC lines for the expression of different SLCs by reverse transcription-PCR and TaqMan real-time PCR. In two of five RCC lines, A498 and 7860, we observed a highly significant expression of SLC22A3 (hOCT3). Uptake of the organic cation [(3)H]MPP (4-methyl-pyridinium iodide) into these cells and also into hOCT3 stably transfected Chinese hamster ovary (CHO) cells was inhibited by irinotecan, vincristine, and melphalan. The K(i) values [determined from Dixon plots] for irinotecan, vincristine, and melphalan were 1.72 +/- 0.45 micromol/L, 17 +/- 4.81 micromol/L, and 366 +/- 51 micromol/L, respectively. Cytotoxic activities of the selected drugs were tested by [(3)H]thymidine incorporation and 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide assays on CHO-hOCT3, A498 (high expression of hOCT3), and ACHN cell lines (low expression of hOCT3). The growth of CHO-hOCT3 was inhibited by 20% more with irinotecan and by 50% more with vincristine compared with nontransfected CHO cells. Melphalan produced 20% to 30% more inhibition in hOCT3-expressing cells compared with nonexpressing control cells. Similar results were obtained for A498 and ACHN cells. Thus, our data support the hypothesis that the sensitivity of tumor cells to chemotherapeutic treatment depends on the expression of transporter proteins mediating specific drug accumulation into target cells. PMID- 19190343 TI - TMPRSS2-ERG gene fusion is not associated with outcome in patients treated by prostatectomy. AB - A significant number of prostate cancers have been shown to have recurrent chromosomal rearrangements resulting in the fusion of the androgen-regulated TMPRSS2 promoter to a member of the ETS transcription factor family, most commonly ERG. This results in ERG overexpression, which may have a direct causal role in prostate tumorigenesis or progression. However, the clinical significance of the rearrangement is unclear, and in particular, relationship to outcome has been inconsistent in recent reports. We analyzed TMPRSS2-ERG gene rearrangement status by fluorescence in situ hybridization in 521 cases of clinically localized surgically treated prostate cancer with 95 months of median follow-up and also in 40 unmatched metastases. Forty-two percent of primary tumors and 40% of metastases had rearrangements. Eleven percent had copy number increase (CNI) of the TMPRRS2-ERG region. Rearrangement alone was associated with lower grade, but not with stage, biochemical recurrence, metastases, or death. CNI with and without rearrangement was associated with high grade and advanced stage. Further, a subgroup of cancers with CNI and rearrangement by deletion, with two or more copies of the deleted locus, tended to be more clinically aggressive. DNA index assessment revealed that the majority of tumors with CNI of TMPRSS2-ERG had generalized aneuploidy/tetraploidy in contrast to tumors without TMPRSS2-ERG CNI, which were predominantly diploid. We therefore conclude that translocation of TMPRSS2-ERG is not associated with outcome, and the aggressive clinical features associated with CNI of chromosome 21 reflect generalized aneuploidy and are not due to CNI specifically of rearranged TMPRSS2-ERG. PMID- 19190344 TI - Association of variants in two vitamin e transport genes with circulating vitamin e concentrations and prostate cancer risk. AB - Significant reductions in prostate cancer incidence and mortality were observed in men randomized to receive 50 mg supplemental vitamin E (alpha-tocopherol) per day in the Alpha-Tocopherol, Beta-Carotene Cancer Prevention (ATBC) Study. We hypothesized that variation in key vitamin E transport genes might directly affect prostate cancer risk or modify the effects of vitamin E supplementation. Associations between prostate cancer risk and 13 polymorphisms in two genes, TTPA and SEC14L2, were examined in 982 incident prostate cancer cases and 851 controls drawn from the ATBC Study. There was no association between the genetic variants and prostate cancer risk. Significant interactions were observed, however, between two variants in SEC14L2 (IVS11+931A>G and IVS11-896A>T) and the trial alpha-tocopherol supplement such that vitamin E supplementation reduced prostate cancer risk among men who were homozygous for either common allele [odds ratios (OR) and 95% confidence intervals (95% CI), 0.52 (0.30-0.90) and 0.64 (0.46 0.88), respectively] and nonsignificantly increased risk among those who carried one or two copies of either variant allele [ORs and 95% CIs, 1.27 (0.90-1.79) and 1.21 (0.96-1.52), respectively; both P for interaction < 0.05]. Genotype phenotype analyses revealed significant but modest differences in baseline circulating concentrations of alpha-tocopherol and serum responses to the vitamin E supplementation for several polymorphisms. This study shows that genetic variation in TTPA and SEC14L2 is associated with serum alpha-tocopherol but does not have a direct effect on prostate cancer. Our results do, however, suggest that polymorphisms in SEC14L2 may modify the effect of vitamin supplementation regimens on prostate cancer risk. PMID- 19190346 TI - Methylation-mediated repression of GADD45alpha in prostate cancer and its role as a potential therapeutic target. AB - Defects in apoptotic pathway contribute to uncontrolled proliferation of cancer cells and confer resistance to chemotherapy. Growth arrest and DNA damage inducible, alpha (GADD45alpha) is up-regulated on docetaxel treatment and may contribute to docetaxel-mediated cytotoxicity. We examined the mechanism of regulation of GADD45alpha in prostate cancer cells and the effect of its up regulation on sensitivity to docetaxel chemotherapy. Expression of GADD45alpha in PC3 cells was higher than that in Du145 and LNCaP cells (17- and 12-fold, respectively; P < 0.05). Although the proximal promoter region was unmethylated in all three cell lines, methylation of a 4 CpG region upstream of the proximal promoter correlated inversely with gene expression levels. Methylation was reversed by treatment of Du145 and LNCaP cells with DNA methyltransferase inhibitors, leading to reactivation of GADD45alpha expression in these cells. The 5' 4 CpG region was also frequently methylated in prostate cancer tissues. Methylation of this region correlated inversely with gene expression in prostate cancer and benign prostate tissues. The methyl binding protein MeCP2 was associated with the methylated 4 CpGs in Du145 cells, and knockdown of MeCP2 in these cells (Du145 MeCP2(-)) led to a significantly increased expression of GADD45alpha (3-fold; P = 0.035) without affecting the methylation status of the gene. Enhanced sensitivity to docetaxel was observed by up-regulation of GADD45alpha in Du145 cells by recombinant expression of GADD45alpha or pretreatment with 5-azacytidine. Our results show that GADD45alpha is epigenetically repressed and is a potential target for treatment of prostate cancer. PMID- 19190345 TI - Epidermal growth factor receptor signaling synergizes with Hedgehog/GLI in oncogenic transformation via activation of the MEK/ERK/JUN pathway. AB - Persistent activation of the Hedgehog (HH)/GLI signaling pathway has been implicated in the development of a number of human cancers. The GLI zinc finger transcription factors act at the end of the HH signaling cascade to control gene expression, and recent studies have shown that the activity of GLI proteins can be additionally modified by integration of distinct signals, such as the MEK/extracellular signal-regulated kinase (ERK) and phosphinositide-3 kinase (PI3K)/AKT pathway. However, little is known about the identity of the upstream activators of these HH/GLI interacting signaling pathways in cancer. Here, we provide evidence that integration of the HH/GLI and epidermal growth factor receptor (EGFR) pathway synergistically induces oncogenic transformation, which depends on EGFR-mediated activation of the RAS/RAF/MEK/ERK but not of the PI3K/AKT pathway. EGFR/MEK/ERK signaling induces JUN/activator protein 1 activation, which is essential for oncogenic transformation, in combination with the GLI activator forms GLI1 and GLI2. Furthermore, pharmacologic inhibition of EGFR and HH/GLI efficiently reduces growth of basal cell carcinoma (BCC) cell lines derived from mice with activated HH/GLI signaling. The results identify the synergistic integration of GLI activator function and EGFR signaling as a critical step in oncogenic transformation and provide a molecular basis for therapeutic opportunities relying on combined inhibition of the HH/GLI and EGFR/MEK/ERK/JUN pathway in BCC. PMID- 19190348 TI - Antitumoral immune response by recruitment and expansion of dendritic cells in tumors infected with telomerase-dependent oncolytic viruses. AB - Virotherapy can potentially be used to induce tumor-specific immune responses and to overcome tumor-mediated tolerance mechanisms because apoptotic tumor cells are exposed together with viral danger signals during oncolysis. However, insufficient numbers of dendritic cells (DC) present at the site of oncolysis can limit a tumor-specific immune response and the resulting therapeutic benefit. We investigated MHC class I peptide-specific immune responses against model antigens ovalbumin (OVA) and hemagglutinin (HA) in mouse tumor models that support efficient replication of the oncolytic adenovirus hTert-Ad. Virotherapy resulted in peptide-specific cytotoxic T-cell responses against intracellular tumor antigens. Triggering of DC and T-cell infiltration to the oncolytic tumors by macrophage inflammatory protein 1alpha (MIP-1alpha, CCL3) and Fms-like tyrosine kinase-3 ligand (Flt3L) enhanced both antitumoral and antiviral immune responses. Although immune-mediated clearance of the virus can restrict therapeutic efficacy of virotherapy, MIP-1alpha/FLT3L-augmented hTert-Ad virotherapy inhibited local tumor growth more effectively than virotherapy alone. In agreement with the hypothesis that immune-mediated mechanisms account for improved outcome in MIP 1alpha/FLT3L virotherapy, we observed systemic antitumoral effects by MIP 1alpha/FLT3L virotherapy on uninfected lung metastasis in immunocompetent mice but not in nude mice. Furthermore, MIP-1alpha/FLT3L virotherapy of primary tumors was strongly synergistic with tumor DC vaccination in inhibition of established lung metastasis. Combined viroimmunotherapy resulted in long-term survival of 50% of treated animals. In summary, improvement of cross-presentation of tumor antigens by triggering of DC and T-cell infiltration during virotherapy enhances antitumoral immune response that facilitates an effective viroimmunotherapy of primary tumors and established metastases. PMID- 19190347 TI - Insulin receptor substrate-2 mediated insulin-like growth factor-I receptor overexpression in pancreatic adenocarcinoma through protein kinase Cdelta. AB - Pancreatic adenocarcinoma (PCA) is an almost invariably fatal disease. Recently, it has been shown by several groups as well as ours that insulin-like growth factor-I receptor (IGF-IR) overexpression is related to higher proliferation, survival, angiogenesis, and highly invasive pancreatic tumors. Several studies have been carried out to understand the pathways that lead to growth factor mediated signaling, but the molecular mechanism of receptor overexpression remains mostly unknown. Treatment with neutralizing antibodies or a specific kinase inhibitor against IGF-IR could block the receptor expression in PCA cells. Furthermore, we also showed that insulin receptor substrate (IRS)-2, but not IRS 1, is involved in regulation of IGF-IR expression, which is most likely not transcriptional control. By blocking mammalian target of rapamycin (mTOR) pathway with rapamycin as well as other biochemical analysis, we defined a unique regulation of IGF-IR expression mediated by protein kinase Cdelta (PKCdelta) and mTOR pathway. Moreover, we showed that the down-regulation of IGF-IR expression due to IRS-2 small interfering RNA can be compensated by overexpression of dominant-active mutant of PKCdelta, suggesting that PKCdelta is downstream of IGF IR/IRS-2 axis. Overall, these findings suggest a novel regulatory role of IRS-2 on the expression of IGF-IR through PKCdelta and mTOR in pancreatic cancer cells. PMID- 19190350 TI - 17Beta-hydroxysteroid dehydrogenase type 12 in human breast carcinoma: a prognostic factor via potential regulation of fatty acid synthesis. AB - 17beta-Hydroxysteroid dehydrogenase type 12 (17beta-HSD12) has been shown to be involved in elongation of very long chain fatty acid (VLCFA) as well as in biosynthesis of estradiol (E2). 17beta-HSD12 expression was also reported in breast carcinomas but its functions have remained unknown. In this study, we examined the correlation between mRNA expression profiles determined by microarray analysis and tissue E2 concentrations obtained from 16 postmenopausal breast carcinoma cases. No significant correlations were detected between 17beta HSD12 expression and E2 concentration. We then immunolocalized this enzyme in 110 cases of invasive ductal carcinoma. 17beta-HSD12 immunoreactivity in breast carcinoma cells was significantly associated with poor prognosis of the patients. We further examined the biological significance of 17beta-HSD12 using cell-based studies. Small interfering RNA-mediated knockdown of 17beta-HSD12 in SK-BR-3 (estrogen receptor-negative breast carcinoma cell line) resulted in significant growth inhibition, which was recovered by the addition of VLCFAs such as arachidonic acid. The status of 17beta-HSD12 immunoreactivity was also correlated with adverse clinical outcome in cyclooxygenase 2 (COX2)-positive breast cancer patients but not in COX2-negative patients. Therefore, these findings indicated that 17beta-HSD12 was not necessarily related to intratumoral E2 biosynthesis, at least in human breast carcinoma, but was rather correlated with production of VLCFAs such as arachidonic acid, which may subsequently be metabolized to prostaglandins by COX2 and result in tumor progression of the patients. PMID- 19190349 TI - Trastuzumab reverses letrozole resistance and amplifies the sensitivity of breast cancer cells to estrogen. AB - In this study, we investigated adaptive mechanisms associated with aromatase inhibitor (AI) resistance in breast cancer cells and show that sensitivity to AIs can be extended through dual inhibition of estrogen receptor (ER) and human epidermal receptor-2 (Her-2) signaling. We used human ER-positive breast cancer cells stably transfected with the aromatase gene (MCF-7Ca). These cells grow as tumors in nude mice and are inhibited by AIs. Despite continued treatment, tumors eventually become insensitive to AI letrozole. The cells isolated from these long term letrozole-treated tumors (LTLT-Ca) were found to have decreased ERalpha levels. Our results suggest that LTLT-Ca cells survive estrogen deprivation by activation of Her-2/mitogen-activated protein kinase (MAPK) pathway. Here, we show that trastuzumab (antibody against Her-2; IC(50) = 0.4 mg/mL) was very effective in restoring the ERalpha levels and sensitivity of LTLT-Ca cells to endocrine therapy by down-regulation of Her-2/MAPK pathway and up-regulation of ERalpha. In contrast, trastuzumab was ineffective in the parental hormone responsive MCF-7Ca cells (IC(50) = 4.28 mg/mL) and xenografts. By blocking Her-2, trastuzumab also up-regulates ERalpha and aromatase expression and hypersensitized MCF-7Ca cells to E(2). We show that trastuzumab is beneficial in hormone-refractory cells and xenografts by restoring ER, implicating Her-2 as a negative regulator of ERalpha. In xenograft studies, the combination of trastuzumab plus letrozole is equally effective in inhibiting growth of MCF-7Ca tumors as letrozole alone. However, on the acquisition of resistance and increased Her-2 expression, the combination of letrozole plus trastuzumab provided superior benefit over letrozole or trastuzumab alone. PMID- 19190351 TI - Sensitivity modeling study for an ozone occurrence during the 1996 Paso Del Norte Ozone Campaign. AB - Surface ozone pollution has been a persistent environmental problem in the US and Europe as well as the developing countries. A key prerequisite to find effective alternatives to meeting an ozone air quality standard is to understand the importance of local anthropogenic emissions, the significance of biogenic emissions, and the contribution of long-range transport. In this study, an air quality modeling system that includes chemistry and transport, CMAQ, an emission processing model, SMOKE, and a mesoscale numerical meteorological model, WRF, has been applied to investigate an ozone event occurring during the period of the 1996 Paso del Norte Ozone Campaign. The results show that the modeling system exhibits the capability to simulate this high ozone occurrence by providing a comparable temporal variation of surface ozone concentration at one station and to capture the spatial evolution of the event. Several sensitivity tests were also conducted to identify the contributions to high surface ozone concentration from eight VOC subspecies, biogenic VOCs, anthropogenic VOCs and long-range transportation of ozone and its precursors. It is found that the reductions of ETH, ISOP, PAR, OLE and FORM help to mitigate the surface ozone concentration, and like anthropogenic VOCs, biogenic VOC plays a nonnegligible role in ozone formation. But for this case, long-range transport of ozone and its precursors appears to produce an insignificant contribution. PMID- 19190352 TI - Human health risk assessment due to global warming--a case study of the Gulf countries. AB - Accelerated global warming is predicted by the Intergovernmental Panel on Climatic Change (IPCC) due to increasing anthropogenic greenhouse gas emissions. The climate changes are anticipated to have a long-term impact on human health, marine and terrestrial ecosystems, water resources and vegetation. Due to rising sea levels, low lying coastal regions will be flooded, farmlands will be threatened and scarcity of fresh water resources will be aggravated. This will in turn cause increased human suffering in different parts of the world. Spread of disease vectors will contribute towards high mortality, along with the heat related deaths. Arid and hot climatic regions will face devastating effects risking survival of the fragile plant species, wild animals, and other desert ecosystems. The paper presents future changes in temperature, precipitation and humidity and their direct and indirect potential impacts on human health in the coastal regions of the Gulf countries including Yemen, Oman, United Arab Emirates, Qatar, and Bahrain. The analysis is based on the long-term changes in the values of temperature, precipitation and humidity as predicted by the global climatic simulation models under different scenarios of GHG emission levels. Monthly data on temperature, precipitation, and humidity were retrieved from IPCC databases for longitude 41.25 degrees E to 61.875 degrees E and latitude 9.278 degrees N to 27.833 degrees N. Using an average of 1970 to 2000 values as baseline, the changes in the humidity, temperature and precipitation were predicted for the period 2020 to 2050 and 2070 to 2099. Based on epidemiological studies on various diseases associated with the change in temperature, humidity and precipitation in arid and hot regions, empirical models were developed to assess human health risk in the Gulf region to predict elevated levels of diseases and mortality rates under different emission scenarios as developed by the IPCC.The preliminary assessment indicates increased mortality rates due to cardiovascular and respiratory illnesses, thermal stress, and increased frequency of infectious vector borne diseases in the region between 2070 and 2099. PMID- 19190353 TI - Pathogenic microorganisms associated with childhood diarrhea in low-and-middle income countries: case study of Yaounde - Cameroon. AB - Notwithstanding significant advancement in the understanding of pathogenesis and management, diarrheal illnesses remain one of the principal causes of global childhood mortality and morbidity. Infections account for most illnesses, with pathogens employing ingenious mechanisms to establish disease. In 2002, an interdisciplinary program "Populations et al. Espaces a Risques SANitaires" (PERSAN) was set up under the patronage of the Development Research Institute (IRD). Focused on health in Cameroon's urban environment, the program mainly sought to identify diarrhea risk factors in Yaounde. So for, a cross-sectional epidemiological study in children aged 6-59 months was carried out using a standardized protocol. The survey was initiated in 2002 and conducted during April to June in the year 2005. 3,034 stool samples were collected from children in twenty neighbourhoods in Yaounde and examined at the Epidemiology and Public Health Laboratory of the Cameroon Pasteur Institute. About 60% of the patients were aged less than two years and 52% were male. Among the 437 patients with the diarrheal disease, 260 were found to be of infectious etiology, i.e. micro organism was detected in 59.5% of the cases. Out of which, 10 (03.8%), 96 (36.9%), and 154 (59.2%) were respectively caused by pathogenic viruses, pathogenic bacteria and pathogenic parasites. Higher prevalence was found in overcrowded and under supply spontaneous settlement (78.4%) than in less crowded and formal residential settlement (21.5%). Etiologic data on diarrheal diseases and their spatial distribution are important tools for public health management and control strategic planning. PMID- 19190354 TI - Gender specific differences in the pros and cons of smoking among current smokers in Eastern Kentucky: implications for future smoking cessation interventions. AB - This study investigated gender differences in the perceived "pros" and "cons" of smoking using the constructs of decisional balance (DB) and stage of change from the Transtheoretical Model. The population distribution for stage of change among a population-based, cross-sectional survey of 155 current smokers over 40 years was: precontemplation (22.6%), contemplation (41.9%), preparation (35.5%). Results of stepwise regression models indicated significant gender differences in DB were in the preparation stage of change; scores on the DB measure increased 3.94 points (95% CI: 1.94, 5.93) for male smokers. Interventions targeting the "pros" and "cons" of smoking may need to be gender specific. PMID- 19190355 TI - Food intakes by preschool children in Flanders compared with dietary guidelines. AB - The objective of this study was to compare food group intakes among preschool children with food-based dietary guidelines (FBDG) and to determine the proportion of children meeting these guidelines. Food consumption of preschool children (2.5-6.5 years) living in Flanders (Belgium) were assessed in a cross sectional study, using proxy reported 3d estimated dietary records (EDR) (n 696). Statistical modelling was used to account for within-individual variation in the 3d EDR. Mean daily intakes of most food groups (beverages, vegetables, fruit and milk) were below the minimum recommendations. Only 'grains and potatoes' and 'meat products' were in line with the recommendations and 'bread and cereals' showed borderline intakes. Mean intakes of energy-dense and low-nutritious foods, which are discouraged within a healthy diet (like snacks and sugared drinks), were high. Furthermore, the percentage of children complying with the different food-based dietary guidelines was for most food groups extremely low (ranging from approximately 4% for fluid and vegetable intakes up to 99% for potato intakes). Boys had in general higher mean intakes of the recommended food groups. In conclusion, preschool children in Flanders follow eating patterns that do not meet Flemish FBDG. Although the impact of these eating habits on preschooler's current and future health should be further investigated, it is clear that nutrition education and intervention are needed among preschool children and their parents in Flanders. PMID- 19190356 TI - Towards integration of environmental and health impact assessments for wild capture fishing and farmed fish with particular reference to public health and occupational health dimensions. AB - The paper offers a review and commentary, with particular reference to the production of fish from wild capture fisheries and aquaculture, on neglected aspects of health impact assessments which are viewed by a range of international and national health bodies and development agencies as valuable and necessary project tools. Assessments sometimes include environmental health impact assessments but rarely include specific occupational health and safety impact assessments especially integrated into a wider public health assessment. This is in contrast to the extensive application of environmental impact assessments to fishing and the comparatively large body of research now generated on the public health effects of eating fish. The value of expanding and applying the broader assessments would be considerable because in 2004 the United Nations Food and Agriculture Organization reports there were 41,408,000 people in the total 'fishing' sector including 11,289,000 in aquaculture. The paper explores some of the complex interactions that occur with regard to fishing activities and proposes the wider adoption of health impact assessment tools in these neglected sectors through an integrated public health impact assessment tool. PMID- 19190357 TI - Cigarette smoking among adolescents in Northwest Ohio: correlates of prevalence and age at onset. AB - This study examined the prevalence and correlates of smoking initiation among adolescents. We have used data from adolescents (n=5,392) ages 10-18 who participated in the 2003 Tobacco Survey, a representative sample of adolescents in Northwest Ohio. A self-report of cigarette smoking was obtained using a questionnaire administered in classrooms. Data were analyzed using weighted chi square and multiple logistic regressions in SAS that accounted for the survey design. The prevalence rates for adolescents that ever tried smoking were 7.4% in elementary (grades 4-5); 17.7% in middle (grades 6-8), and 41.4% in high (grades 9-12) schools, respectively. The highest prevalence rate was among Hispanics. Having a close friend that smoked and a smoker at home correlated significantly with both initiation of smoking and smoking at an earlier age. Smoking was correlated with low academic achievement among adolescents in all grades. Students who reported smoking by parents or siblings were significantly more likely to start smoking at an earlier age, compared to other students living in a non-smoking home environment. Smoking prevention program should include components focused on adolescents' home environment and should start as early as the 4th grade. PMID- 19190358 TI - Web 3D for public, environmental and occupational health: early examples from second life. AB - Over the past three years (2006-2008), the medical/health and public health communities have shown a growing interest in using online 3D virtual worlds like Second Life(R) (http://secondlife.com/) for health education, community outreach, training and simulations purposes. 3D virtual worlds are seen as the precursors of 'Web 3D' , the next major iteration of the Internet that will follow in the coming years. This paper provides a tour of several flagship Web 3D experiences in Second Life(R), including Play2Train Islands (emergency preparedness training), the US Centers for Disease Control and Prevention--CDC Island (public health), Karuna Island (AIDS support and information), Tox Town at Virtual NLM Island (US National Library of Medicine--environmental health), and Jefferson's Occupational Therapy Center. We also discuss the potential and future of Web 3D. These are still early days of 3D virtual worlds, and there are still many more untapped potentials and affordances of 3D virtual worlds that are yet to be explored, as the technology matures further and improves over the coming months and years. PMID- 19190359 TI - Treatment vault shielding for a flattening filter-free medical linear accelerator. AB - The requirements for shielding a treatment vault with a Varian Clinac 2100 medical linear accelerator operated both with and without the flattening filter were assessed. Basic shielding parameters, such as primary beam tenth-value layers (TVLs), patient scatter fractions, and wall scatter fractions, were calculated using Monte Carlo simulations of 6, 10 and 18 MV beams. Relative integral target current requirements were determined from treatment planning studies of several disease sites with, and without, the flattening filter. The flattened beam shielding data were compared to data published in NCRP Report No. 151, and the unflattened beam shielding data were presented relative to the NCRP data. Finally, the shielding requirements for a typical treatment vault were determined for a single-energy (6 MV) linac and a dual-energy (6 MV/18 MV) linac. With the exception of large-angle patient scatter fractions and wall scatter fractions, the vault shielding parameters were reduced when the flattening filter was removed. Much of this reduction was consistent with the reduced average energy of the FFF beams. Primary beam TVLs were reduced by 12%, on average, and small-angle scatter fractions were reduced by up to 30%. Head leakage was markedly reduced because less integral target current was required to deliver the target dose. For the treatment vault examined in the current study, removal of the flattening filter reduced the required thickness of the primary and secondary barriers by 10-20%, corresponding to 18 m(3) less concrete to shield the single energy linac and 36 m(3) less concrete to shield the dual-energy linac. Thus, a shielding advantage was found when the linac was operated without the flattening filter. This translates into a reduction in occupational exposure and/or the cost and space of shielding. PMID- 19190361 TI - Cramer-Rao lower bound of basis image noise in multiple-energy x-ray imaging. AB - We present an analytical method to compute the basis image noise in the context of multi-energy x-ray imaging based on the Cramer-Rao lower bound (CRLB). The proposed formalism extends the original idea of Alvarez and Macovski (1976 Phys. Med. Biol. 21 733) to estimate the noise in the photo-effect and Compton-effect basis images in the case of dual-energy imaging. It includes an arbitrary number of independent, spectrally distinct attenuation measurements and also goes beyond the two-dimensional decomposition of the attenuation, including, e.g., a contrast agent as a third basis material. To illustrate our method, we consider three simple applications. The first application is to study the influence of the exact values for the energy thresholds on the basis image noise for a binned photon counting system. The second application relates to the same detector system as the first and is an investigation of the dependence of the basis image noise on the energy resolution of the detector system. Finally, the third application provides an example for the case of an energy-integrating detector: the aim is to optimize the front-scintillator layer thickness of a dual-crystal detector for dual-energy imaging. The CRLB is used to minimize the noise of a photo effect/Compton-effect basis material decomposition. PMID- 19190360 TI - A high-sensitivity small animal SPECT system. AB - Medical imaging using single gamma-ray-emitting radionuclides typically makes use of parallel hole collimators or pinholes in order to achieve good spatial resolution. However, a tradeoff in sensitivity is inherent in the use of a collimator, and modern preclinical single photon emission computed tomography (SPECT) systems detect a very small fraction of emitted gamma rays, often less than 0.1%. A system for small animal SPECT imaging which uses no collimators could potentially achieve very high sensitivity-several tens of percent-with reasonably sized detectors. This would allow two significant improvements in preclinical studies: images could be obtained more rapidly, allowing higher throughput for screening applications, or for dynamic processes to be observed with very good time resolution; and images could be obtained with less radioactive tracer, making possible the in vivo imaging of low-capacity receptor systems, aiding research into new tracer compounds, and reducing the cost and easing the regulatory burden of an experiment. Of course, a system with no collimator will not be able to approach the submillimeter spatial resolutions produced by the most advanced pinhole and collimated systems, but a high sensitivity system with resolution of order 1 cm could nonetheless find significant and new use in the many molecular imaging applications which do not require good spatial resolution-for example, screening applications for drug development or new imaging agents. Rather than as an alternative to high resolution SPECT systems, the high-sensitivity system is proposed as a radiotracer alternative to optical imaging for small animals. We have developed a prototype system for mouse imaging applications. The scanner consists of two large, thin, closely spaced scintillation detectors. Simulation studies indicate that a FWHM spatial resolution of 7 mm is possible. In an in vivo mouse imaging study using the (99m)Tc labeled tracer MAG-3, the sensitivity of the system is measured to be 40%. Simple projection images created by analytically combining the two detectors' data show sufficient resolution to observe the dynamic distribution of the radiotracer in the mouse. PMID- 19190362 TI - Megavoltage image contrast with low-atomic number target materials and amorphous silicon electronic portal imagers. AB - Low-atomic number (Z) targets have been shown to improve contrast in megavoltage (MV) images when using film-screen detection systems. This research aims to quantify the effect of low-Z targets on MV image contrast using an amorphous silicon electronic portal image detector (a-Si EPID) through both experimental measurement and Monte Carlo (MC) simulation. Experimental beams were produced with the linac running in the 6 MeV electron mode and with a 1.0 cm aluminum (Al, Z = 13) target replacing flattening filtration in the carousel, (6 MeV/Al). A 2100EX Varian linac equipped with an aS500 EPID was used with the QC3 MV phantom for the majority of contrast measurements. The BEAMnrc/EGSnrc MC package was used to build a model of the full imaging system including beam generation (linac head), the a-Si detector and the contrast phantom. The model accurately reproduces contrast measurements to within 2.5% for both the standard 6 MV therapy beam and the 6 MeV/Al beam. The contrast advantage of 6 MeV/Al over 6 MV, as quantified with the QC3 phantom, ranged from a factor increase of 1.6 +/- 0.1 to 2.8 +/- 0.2. Only a modest improvement in contrast was seen when the incident electron energy was reduced to 4 MeV (up to factor of 1.2 +/- 0.1 over 6 MeV/Al) or with removal of the copper build-up layer in the detector, (up to factor of 1.2 +/- 0.1 over 6 MeV/Al). Further decreasing the target Z, to beryllium (Be, Z = 4), at 4 MeV showed no significant improvement over 4 MeV/Al. Experimentally, the contrast advantage of 6 MeV/Al over 6 MV was found to decrease with increasing patient thickness, as can be expected due to selective attenuation of low-energy photons. At head and neck-like thicknesses, the low-Z advantage is a factor increase of 1.7 +/- 0.1. PMID- 19190363 TI - {[SmCl(en)4][In2Te4]}n: an indium telluride with a one-dimensional sinusoidal chain. AB - The title compound, catena-poly[[chloridotetrakis(ethylenediamine kappa(2)N,N')samarium(III)] [indium(III)-di-mu-tellurido-indium(III)-di-mu tellurido]], {[SmCl(C(2)H(8)N(2))(4)][In(2)Te(4)]}(n), consists of a one dimensional sinusoidal {[InTe(2)](-)}(n) anionic chain and [SmCl(en)(4)](2+) cations (en is ethylenediamine). The only other previously reported lanthanide analogue, viz. [LaCl(en)(4)][In(2)Te(4)], contains more usual linear one dimensional {[InTe(2)](-)}(n) anion chains [Chen, Li, Chen & Proserpio (1998). Inorg. Chim. Acta, 273, 255-258]. The one-dimensional polymeric {[InTe(2)](-)}(n) chain is built of InTe(4) tetrahedra sharing opposite edges. The Sm(III) ion in the [SmCl(en)(4)](2+) cation is nine-coordinated by eight N atoms from four bidentate en molecules and by one chloride ion to form a monocapped square antiprismatic geometry. The presence or absence of N-H...Cl hydrogen bonding is shown to affect the conformation of the anion. PMID- 19190364 TI - Poly[triaqua(mu4-4-carboxybenzenesulfonato-kappa4O:O':O'':O''')(4 carboxybenzenesulfonato-kappaO)strontium(II)]. AB - This study presents the coordination modes and two-dimensional network of a novel strontium(II) coordination polymer, [Sr(C(7)H(5)O(5)S)(2)(H(2)O)(3)](n). The eight-coordinate Sr(2+) ion is in a distorted bis-disphenoidal coordination environment, surrounded by four sulfonate and one carboxyl O atom from five benzenesulfonate ligands, two of which are symmetry unique, and by three O atoms from three independent aqua ligands. The compound exhibits a monolayer structure with coordination bonds within and hydrogen bonds between the layers. The mu(4) acid ligand bridges the metal ions in two dimensions to form a thick undulating monolayer with a hydrophobic interior and hydrophilic surfaces. A second independent monoanion is arranged outward from both sides of the monolayer and serves to link adjacent monolayers via carboxyl-water and water-carboxyl hydrogen bonds. PMID- 19190365 TI - Poly[[[mu-1,1'-(butane-1,4-diyl)diimidazole-kappa2N:N'](mu-cyclohexane-1,4 dicarboxylato-kappa4O1,O1':O4,O4')cadmium(II)] hemihydrate]: a parallel interpenetrating two-dimensional (4,4) network. AB - In the title coordination compound, {[Cd(C(8)H(10)O(4))(C(10)H(14)N(4))].0.5H(2)O}(n), the 1,1'-(butane-1,4 diyl)diimidazole ligand and the cyclohexane-1,4-dicarboxylate dianion both function in a bridging mode to link adjacent cadmium(II) centers into a two dimensional four-connected (4,4) network. The networks are parallel to the (001) plane. Two (4,4) networks are interpenetrated in an unusual parallel mode. The compound is the first two-dimensional parallel interpenetrating (4,4) network structure based on a flexible dicarboxylate and a long bidentate N-donor ligand. The interpenetrating nets are further consolidated by water-carboxylate O-H...O hydrogen bonds. PMID- 19190366 TI - trans-Chlorido(phenyl)bis(triphenylphosphine)nickel(II) and its 1:1 cocrystal with chloridobis(triphenylphosphine)nickel(I). AB - Two conformational polymorphs of trans chlorido(phenyl)bis(triphenylphosphine)nickel(II), [Ni(C(6)H(5))Cl(C(18)H(15)P)(2)], (1), viz. orange needle-shaped crystals (form I) and brown prism-shaped crystals (form II), were obtained under different crystallization conditions from a mixture of toluene and n-hexane, and characterized by single-crystal X-ray diffraction at low temperature. These two forms were compared with that published previously [Zeller, Herdtweck & Strassner (2003). Eur. J. Inorg. Chem. pp. 1802-1806], characterized at room temperature. Additionally, blue-green prisms of a 1:1 cocrystal of complex (1) with chloridobis(triphenylphosphine)nickel(I), (2), viz. trans chlorido(phenyl)bis(triphenylphosphine)nickel(II) chloridobis(triphenylphosphine)nickel(I) (1/1), [Ni(C(6)H(5))Cl(C(18)H(15)P)(2)].[NiCl(C(18)H(15)P)(2)], (3), were obtained concomitantly with form I. In forms I and II, as well as in the cocrystal, the overall crystal packings are determined by an energetic interplay between intramolecular torsions and weak intermolecular C-H...pi and C-H...Cl interactions. PMID- 19190367 TI - A two-dimensional network formed by self-associating silver(I) perchlorate with 3 [4-(2-thienyl)-2H-cyclopenta[d]pyridazin-1-yl]benzonitrile. AB - In the organometallic silver(I) supramolecular complex poly[[silver(I)-mu(3)-3-[4 (2-thienyl)-2H-cyclopenta[d]pyridazin-1-yl]benzonitrile] perchlorate methanol solvate], {[Ag(C(18)H(11)N(3)S)](ClO(4)).CH(3)OH}(n), there is only one type of Ag(I) center, which lies in an {AgN(2)Spi} coordination environment. Two unsymmetric multidentate 3-[4-(2-thienyl)-2H-cyclopenta[d]pyridazin-1 yl]benzonitrile (L) ligands link two Ag(I) atoms through pi-Ag(I) interactions into an organometallic box-like unit, from which two 3-cyanobenzoyl arms stretch out in opposite directions and bind two Ag(I) atoms from neighboring box-like building blocks. This results in a novel two-dimensional network extending in the crystallographic bc plane. These two-dimensional sheets stack together along the crystallographic a axis to generate parallelogram-like channels. The methanol solvent molecules and the perchlorate counter-ions are located in the channels, where they are fixed by intermolecular hydrogen-bonding interactions. This architecture may provide opportunities for host-guest chemistry, such as guest molecule loss and absorption or ion exchange. The new fulvene-type multidentate ligand L is a good candidate for the preparation of Cp-Ag(I)-containing (Cp is cyclopentadienyl) organometallic coordination polymers or supramolecular complexes. PMID- 19190368 TI - Robust packing in cyclopalladated primary amines: isomorphous crystal structures of four complexes with varying substitution patterns. AB - The crystal structures of (SP-4-4)-[rac-2-(1-aminoethyl)phenyl kappa(2)C(1),N]chlorido(pyridine-kappaN)palladium(II), [Pd(C(8)H(10)N)Cl(C(5)H(5)N)], (I), (SP-4-4)-[rac-2-(1-aminoethyl)phenyl kappa(2)C(1),N]bromido(pyridine-kappaN)palladium(II), [PdBr(C(8)H(10)N)(C(5)H(5)N)], (II), (SP-4-4)-[rac-2-(1-aminoethyl)-5-bromophenyl kappa(2)C(1),N]bromido(4-methylpyridine-kappaN)palladium(II), [PdBr(C(8)H(9)BrN)(C(6)H(7)N)], (III), and (SP-4-4)-[rac-2-(1-aminoethyl)-5 bromophenyl-kappa(2)C(1),N]iodido(4-methylpyridine-kappaN)palladium(II), [Pd(C(8)H(9)BrN)I(C(6)H(7)N)], (IV), are reported. The latter is the first iodide complex in this class of compounds. All four complexes crystallize in the same space group, viz. I4(1)/a, with very similar lattice parameters a and more flexible lattice parameters c. Their packing corresponds to that of their enantiomerically pure congeners, which crystallize in the t2 subgroup I4(1). PMID- 19190369 TI - Luminescence properties of three structures built from 3-cyano-4-dicyanomethylene 5-oxo-4,5-dihydro-1H-pyrrol-2-olate and alkaline metals (Na, K and Rb). AB - The structures of three salts of 3-cyano-4-dicyanomethylene-5-oxo-4,5-dihydro-1H pyrrol-2-olate with alkali metals (Na, K and Rb) are related to their luminescence properties. The Rb salt, rubidium(I) 3-cyano-4-dicyanomethylene-5 oxo-4,5-dihydro-1H-pyrrol-2-olate, Rb(+).C(8)HN(4)O(2)(-), is isomorphous with the previously reported potassium salt. For the Na compound, sodium(I) 3-cyano-4 dicyanomethylene-5-oxo-4,5-dihydro-1H-pyrrol-2-olate dihydrate, Na(+).C(8)HN(4)O(2)(-).2H(2)O, two independent sodium ions, located on inversion centers, are coordinated by four water molecules each and additionally by two cyano groups for one and two carbonyl groups for the other. The luminescence spectra in solution are unaffected by the nature of the cation but vary strongly with the dielectric constant of the solvent. In the solid state, the emission maxima vary with structural features; the redshift of the maximum luminescence varies inversely with the distance between the stacked anions. PMID- 19190370 TI - A three-dimensional homochiral metal-organic framework constructed from manganese(II) with S-carboxymethyl-N-(p-tosyl)-L-cysteine and 4,4'-bipyridine. AB - In the chiral polymeric title compound, poly[aqua(4,4'-bipyridine)[mu(3)-S carboxylatomethyl-N-(p-tosyl)-L-cysteinato]manganese(II)], [Mn(C(12)H(13)NO(6)S(2))(C(10)H(8)N(2))(H(2)O)](n), the Mn(II) ion is coordinated in a distorted octahedral geometry by one water molecule, three carboxylate O atoms from three S-carboxyatomethyl-N-(p-tosyl)-L-cysteinate (Ts-cmc) ligands and two N atoms from two 4,4'-bipyridine molecules. Each Ts-cmc ligand behaves as a chiral mu(3)-linker connecting three Mn(II) ions. The two-dimensional frameworks thus formed are further connected by 4,4'-bipyridine ligands into a three dimensional homochiral metal-organic framework. This is a rare case of a homochiral metal-organic framework with a flexible chiral ligand as linker, and this result demonstrates the important role of noncovalent interactions in stabilizing such assemblies. PMID- 19190371 TI - A novel three-dimensional copper(I) coordination polymer constructed from a copper-bromide net and nicotinic acid ligands. AB - The title compound, poly[mu(3)-bromido-(pyridine-3-carboxylato-kappaN)copper(I)], [CuBr(C(6)H(5)NO(2))](n), is a novel coordination polymer based on a copper bromide net and nicotinic acid ligands. The asymmetric unit contains one copper(I) ion, one bromide ligand and one nicotinic acid ligand, all on general positions. The Cu(I) atom is tetrahedral and coordinated by three bridging Br atoms and the N atom from the nicotinic acid ligand. The Cu-Br units form alternating six-membered chair-patterned rings in net-like layers. The attached nicotinic acid units point alternately up and down. The layers are assembled into a three-dimensional network via intermolecular O-H...O and C-H...Br hydrogen bonding interactions. PMID- 19190372 TI - Square-grid coordination networks of diaquabis(4,4'-bipyridyl)copper(II) crosslinked by hydrogen bonds through two monoanions of 1-benzofuran-2,3 dicarboxylic acid and five molecules of water. AB - The title compound, poly[[[diaquacopper(II)]-di-mu-4,4'-bipyridyl] bis(3-carboxy 1-benzofuran-2-carboxylate) pentahydrate], {[Cu(C(10)H(8)N(2))(2)(H(2)O)(2)](C(10)H(5)O(5))(2).5H(2)O}(n), crystallizes in a single-framework architecture. It is composed of two-dimensional square-grid coordination networks of 1:2:2 copper-4,4'-bipyridine-water units, wherein each copper ion coordinates equatorially to four bipyridyl units and axially to two water ligands. The polymeric nets are intercalated by layers of the benzofurandicarboxylic acid monoanions and additional water species. An extensive array of hydrogen bonds interlinks the various components of the structure. The Cu atom and the bipyridyl entities are located on axes of twofold rotation. This study confirms the preferred monoanionic nature of the benzofurandicarboxylic acid molecule. It reveals a rarely observed extended coordination polymer composed only of copper ions and bipyridyl linkers, and an interesting hydrogen bonding connectivity between the polymeric layers aided by the benzofurandicarboxylic acid and water components intercalated in the structure. PMID- 19190373 TI - A novel two-dimensional framework based on unprecedented cadmium(II) chains. AB - The bent ligand 4-[(1H-1,2,4-triazol-1-yl)methyl]benzoic acid (HL) has been used to create the novel two-dimensional coordination polymer poly[mu(2)-aqua-mu(2) chlorido-{mu(2)-4-[(1H-1,2,4-triazol-1-yl)methyl]benzoato}cadmium(II)], [Cd(C(10)H(8)N(3)O(2))Cl(H(2)O)](n), under hydrothermal reaction of HL with cadmium chloride. The crystallographically unique Cd atom is seven-coordinated in an approximately pentagonal-bipyramidal environment of two carboxylate O atoms, two water O atoms, two Cl atoms and one triazole N atom. A notable feature is the presence of zigzag ...Cd...Cd... inorganic chains, in which neighboring Cd(II) ions are doubly bridged by pairs of mu(2)-Cl atoms and mu(2)-H(2)O ligands in an alternating fashion. To the authors' knowledge, this is the first example containing this bridging mode in a cadmium(II) framework. The chains are connected to one another through the bridging L(-) ligand into a two-dimensional undulating network. All of the two-dimensional nets stack exactly together in an ...AA... stacking sequence along the crystallographic b axis. Neighboring layers are further linked into a three-dimensional framework via interlayer hydrogen bonding interactions. PMID- 19190374 TI - Directed anisotropy in bis(acetato-kappa2O,O')diaquazinc(II) at 110 and 250 K. AB - The molecule of the title compound, [Zn(C(2)H(3)O(2))(2)(H(2)O)(2)], is located on a twofold axis in the crystal structure. The displacement parameters and the thermal expansion of the crystal show significant anisotropy. This is explained by the two-dimensional hydrogen-bonded structure, with only very weak interactions perpendicular to it. Besides the overall molecular motion, there are internal vibrations, which cause the Zn-O(carboxylate) bonds to fail the Hirshfeld rigid-bond test. It is shown that this can be interpreted in terms of the steric strain in the four-membered chelate ring due to the bidentate carboxylate coordination. PMID- 19190375 TI - A three-dimensional Al(III)/Na(I) metal-organic framework resulting from dimethylformamide hydrolysis. AB - The three-dimensional metal-organic framework poly[bis(dimethylammonium) [hexa mu(2)-formato-kappa(12)O:O'-aluminium(III)sodium(I)]], {(C(6)H(8)N)(2)[AlNa(HCOO)(6)]}(n), was obtained serendipitously and has been characterized by X-ray diffraction. The product has arisen as a result of a hydrolysis reaction of dimethylformamide (DMF) and contains dimethylammonium (DMA) cations included in structural voids formed by a three-dimensional [AlNa(HCOO)(6)](-) network. This study provides evidence that, in the presence of traces of aluminium, DMF stored in a glass bottle can be hydrolysed to formate and dimethylamine with simultaneous extraction of Na(+) cations from the glass. It also demonstrates that care must be taken regarding the metal and water content when DMF is not freshly distilled, since the hydrolysis of amide can occur. PMID- 19190376 TI - Two bicyclic dinuclear complexes generated from 3,3'-[1,3,4-thiadiazole-2,5 diyldi(thiomethylene)]dibenzoic acid (L) and dimethylformamide (DMF): [Cu(L)(DMF)]2and [Zn(L)(DMF)]2. AB - A new 1,3,4-thiadiazole bridging ligand, namely 3,3'-[1,3,4-thiadiazole-2,5 diyldi(thiomethylene)]dibenzoic acid (L), has been used to create the novel isomorphous complexes bis{mu-3,3'-[1,3,4-thiadiazole-2,5 diyldi(thiomethylene)]dibenzoato}bis[(N,N-dimethylformamide)copper(II)], [Cu(2)(C(18)H(12)N(2)O(4)S(3))(2)(C(3)H(7)NO)(2)], (I), and bis{mu-3,3'-[1,3,4 thiadiazole-2,5-diyldi(thiomethylene)]dibenzoato}bis[(N,N dimethylformamide)zinc(II)], [Zn(2)(C(18)H(12)N(2)O(4)S(3))(2)(C(3)H(7)NO)(2)], (II). Both exist as centrosymmetric bicyclic dimers constructed through the syn syn bidentate bridging mode of the carboxylate groups. The two rings share a metal-metal bond and each of the metal atoms possesses a square-pyramidal geometry capped by the dimethylformamide molecule. The 1,3,4-thiadiazole rings play a critical role in the formation of a pi-pi stacking system that expands the dimensionality of the structure from zero to one. The thermogravimetric analysis of (I) indicates decomposition of the coordinated ligands on heating. Compared with the fluorescence of L in the solid state, the fluorescence intensity of (II) is relatively enhanced with a slight redshift, while that of (I) is quenched. PMID- 19190377 TI - A novel threefold-interpenetrating primitive cubic network based on a dinuclear Zn2 node. AB - In the mixed-ligand metal-organic polymeric compound poly[[mu(2)-1,4-bis(imidazol 1-yl)benzene](mu(2)-terephthalato)dizinc(II)], [Zn(2)(C(8)H(4)O(4))(2)(C(12)H(10)N(4))](n) or [Zn(2)(bdc)(2)(bib)](n) [H(2)bdc is terephthalic acid and bib is 1,4-bis(imidazol-1-yl)benzene], the asymmetric unit contains one Zn(II) ion, with two half bdc anions and one half bib molecule lying around inversion centers. The Zn(II) ion is in a slightly distorted tetrahedral environment, coordinated by three carboxylate O atoms from three different bdc anions and by one bib N atom. The crystal structure is constructed from the secondary building unit (SBU) [Zn(2)(CO(2))(2)N(2)O(2)], in which the two metal centers are held together by two bdc linkers with bis(syn,syn-bridging bidentate) bonding modes. The SBU is connected by bdc bridges to form a two dimensional grid-like (4,4)-layer, which is further pillared by the bib ligand. Topologically, the dinuclear SBU can be considered to be a six-connected node, and the extended structure exhibits an elongated primitive approximately cubic framework. The three-dimensional framework possesses a large cavity with dimensions of approximately 10 x 13 x 17 A in cross-section. The potential porosity is filled with mutual interpenetration of two identical equivalent frameworks, generating a novel threefold interpenetrating network with an alpha polonium topology [Abrahams, Hoskins, Robson & Slizys (2002). CrystEngComm, 4, 478-482]. PMID- 19190378 TI - Influence of solvent on the structures of two one-dimensional cobalt(II) coordination polymers with tetrachloroterephthalate. AB - The title cobalt coordination polymers, catena-poly[[[aquatripyridinecobalt(II)] mu-tetrachloroterephthalato] pyridine solvate], {[Co(C(8)Cl(4)O(4))(C(5)H(5)N)(3)(H(2)O)].C(5)H(5)N}(n), (I), and catena poly[[[diaquadipyridinecobalt(II)]-mu-tetrachloroterephthalato] 1,4-dioxane trihydrate], {[Co(C(8)Cl(4)O(4))(C(5)H(5)N)(2)(H(2)O)(2)].C(4)H(8)O(2).3H(2)O}(n), (II), have been prepared with tetrachloroterephthalic acid (H(2)BDC-Cl(4)) under different solvent media. Both complexes form infinite cobalt(II)-tetrachloroterephthalate polymeric chains. In (I), two independent Co(II) ions are six-coordinated through N(3)O(3) donor sets in slightly distorted octahedral geometries provided by two carboxylate and three pyridine ligands, and one water molecule. The structure of (II) contains two independent Co(II) atoms, each lying on a twofold axis, which adopt a tetragonally distorted N(2)O(4) octahedral geometry via two carboxylate groups, two pyridine ligands and two water molecules. The different stoichiometry of coordinated and solvent guest molecules leads to different two-dimensional supramolecular networks, with (I) utilizing C-H...pi and weak pi-pi interactions and (II) utilizing mainly conventional hydrogen bonding. PMID- 19190379 TI - An unprecedented three-dimensional silver(I) cluster built up from pyrazine-2,3 dicarboxylate ligands. AB - The title complex, poly[(mu-3-carboxypyrazine-2-carboxylato)(mu-pyrazine-2,3 dicarboxylato)trisilver(I)], [Ag(3)(C(6)H(2)N(2)O(4))(C(6)H(3)N(2)O(4))](n) or [Ag(3)(pzdca)(Hpzdca)](n) (H(2)pzdca is pyrazine-2,3-dicarboxylic acid), has a three-dimensional structure. The carboxylate groups of the pzdca(2-) and Hpzdca( ) ligands adopt both bridging and chelating coordination modes. Although each Ag(I) ion displays a tetrahedral coordination, the coordination environment of each Ag atom is very different, viz. AgN(3)O, AgNO(3) and AgO(4). PMID- 19190380 TI - A polymorph of tetrakis(acetonitrile-kappaN)copper(I) tetrafluoridoborate. AB - A P2(1)2(1)2(1) polymorph of the title compound, [Cu(CH(3)CN)(4)]BF(4), is reported. The crystal structure is very similar to the structure of the Pna2(1) polymorph reported by Jones & Crespo [Acta Cryst. (1998), C54, 18-20]. The anions and one of the three independent cations occupy similar positions in both polymorphs. Two of the four symmetry-related positions of the other two cations are also identical in the two polymorphs, and the other two positions are related by mirror symmetry. The crystal used for the structure determination contained a volume fraction of 0.088 (7) of the Pna2(1) polymorph. PMID- 19190381 TI - Bis[2-(2-hydroxyethyl)pyridinium] mu-decavanadato-bis[pentaaquamanganate(II)] tetrahydrate. AB - The structure of the title compound, (C(7)H(10)NO)(2)[Mn(2)V(10)O(28)(H(2)O)(10)].4H(2)O or (C(5)H(4)NHCH(2)CH(2)OH)(2)[{Mn(H(2)O)(5)}(2)V(10)O(28)].4H(2)O, at 293 (2) K has triclinic (P?overline{1}) symmetry. The asymmetric unit consists of one half of a decavanadate anion of C(i) symmetry, one [Mn(H(2)O)(5)](2+) group, one 2-(2 hydroxyethyl)pyridinium cation and two solvent water molecules. The decavanadate ion bridges between two [Mn(H(2)O)(5)](2+) groups, thus forming a dodecanuclear complex unit. Complex units are connected via a hydrogen-bonding network, forming supramolecular layers lying in the (001) plane. Cations and solvent water molecules are located between these layers. PMID- 19190382 TI - A new polymorph of triphenylmethylamine: the effect of hydrogen bonding. AB - Crystallization of the hexane reaction mixture after treatment of LiGe(OCH(2)CH(2)NMe(2))(3) with Ph(3)CN(3) gives rise to a new triclinic (space group P?overline{1}) polymorph of triphenylmethylamine, C(19)H(17)N, (I), containing dimers formed by N-H...N hydrogen bonds, whereas the structure of the known orthorhombic (space group P2(1)2(1)2(1)) polymorph of this compound, (II), consists of isolated molecules. While the dimers in (I) lie across crystallographic inversion centres, the molecules are not truly related by them. The centrosymmetric structure is due to the statistical disordering of the amino H atoms participating in the N-H...N hydrogen-bonding interactions, and thus the inversion centre is superpositional. The conformations and geometric parameters of the molecules in (I) and (II) are very similar. It was found that the polarity of the solvent does not affect the capability of triphenylmethylamine to crystallize in the different polymorphic modifications. The orthorhombic polymorph, (II), is more thermodynamically stable under normal conditions than the triclinic polymorph, (I). The experimental data indicate the absence of a phase transition in the temperature interval 120-293 K. The densities of (I) (1.235 Mg m(-3)) and (II) (1.231 Mg m(-3)) at 120 K are practically equal. It would seem that either the kinetic factors or the effects of the other products of the reaction facilitating the hydrogen-bonded dimerization of triphenylmethylamine molecules are the determining factor for the isolation of the triclinic polymorph (I) of triphenylmethylamine. PMID- 19190383 TI - Tetrameric aggregate of 1,c-3-diphenyltetran-r-1-ol via an R4(4)8 homodromic ring. AB - The compound 1,c-3-diphenyltetran-r-1-ol (systematic name: 1,c-3-diphenyl-1,2,3,4 tetrahydro-r-1-naphthol), C(22)H(20)O, which possesses the tetrahydronaphthalene core that is found in a large number of natural products, crystallizes with Z' = 4 and with the four molecules forming a hydrogen-bonded cyclic aggregate. The aliphatic six-membered rings are present with two different conformations in the molecules of the asymmetric unit. A comparison with similar fragments reveals their conformational flexibility. In addition, the structure demonstrates the relative stereochemistries of the chiral centers, which are important since the title compound is used in the stereoselective synthesis of compounds with therapeutic activity. PMID- 19190384 TI - gem-Dimethylcyclopropanation of dibenzylideneacetone using triisopropyl sulfoxonium tetrafluoroborate. AB - The reaction between dibenzylideneacetone (dba) and triisopropyl sulfoxonium tetrafluoroborate has been reinvestigated. The stereochemistry of the major diasteromeric bis(gem-dimethylcyclopropane) adduct has now been assigned as [(1RS,3RS)-2,2-dimethyl-3-phenylcyclopropyl][(1SR,3SR)-2,2-dimethyl-3 phenylcyclopropyl]methanone, C(23)H(26)O, by X-ray crystallographic studies on a twinned crystal. The asymmetric unit contains two molecules of the adduct, the conformations of which differ in the orientation of the phenyl ring relative to the adjacent cyclopropanated double bond. The carbonyl groups of each adduct are aligned approximately along the a axis and in opposite directions to each other. The molecules pack to give a sinusoidal pattern along the b axis. This is the first acyclic bis(dimethylcyclopropyl) ketone for which an X-ray crystal structure determination has been reported, and is also the first bis cyclopropanated dba analogue. The knowledge that the major diastereomer has the meso structure (and therefore the confirmation that the minor isomer is the racemate) will prove invaluable in future studies to utilize bis(dimethylcyclopropyl) ketones as reagents, in rearrangement processes, and as potential ligands and ligand precursors in organometallic chemistry. PMID- 19190385 TI - Hydrogen-bonded supramolecular motifs in 2-amino-4,6-dimethoxypyrimidinium picrate and pyrimethaminium picrate dimethyl sulfoxide solvate. AB - In the crystal structures of 2-amino-4,6-dimethoxypyrimidinium 2,4,6 trinitrophenolate (picrate), C(6)H(10)N(3)O(2)(+).C(6)H(2)N(3)O(7)(-), (I), and 2,4-diamino-5-(4-chlorophenyl)-6-ethylpyrimidin-1-ium (pyrimethaminium or PMN) picrate dimethyl sulfoxide solvate, C(12)H(14)ClN(4)(+).C(6)H(2)N(3)O(7)( ).C(2)H(6)OS, (II), the 2-amino-4,6-dimethoxypyrimidine and PMN cations are protonated at one of the pyrimidine N atoms. The picrate anion interacts with the protonated cations through bifurcated N-H...O hydrogen bonds, forming R(2)(1)(6) and R(1)(2)(6) ring motifs. In (I), Z' = 2. In (II), two inversion-related PMN cations are connected through a pair of N-H...N hydrogen bonds involving the 4 amino group and the uncharged N atom of the pyrimidine ring, forming a cyclic hydrogen-bonded R(2)(2)(8) motif. In addition to the pairing, the O atom of the dimethyl sulfoxide solvent molecule bridges the 2-amino and 4-amino groups on both sides of the paired bases, resulting in a self-complementary ...DADA... array of quadruple hydrogen-bonding patterns. PMID- 19190386 TI - Two enantiomerically pure cyclic arenesulfonamide hydrochloride salts. AB - The crystal structures of N-[(1R)-1-(1-naphthyl)ethyl]-3,4-dihydro-2H-1,2 benzothiazin-4-aminium 1,1-dioxide chloride, C(20)H(21)N(2)O(2)S(+).Cl(-), (I), a six-membered cyclic sulfonamide, and (1R)-N-[(5,5-dioxo-6,7 dihydrodibenzo[d,f][1,2]thiazepin-7-yl)methyl]-1-(1-naphthyl)ethanaminium chloride, C(26)H(25)N(2)O(2)S(+).Cl(-), (II), a seven-membered cyclic sulfonamide, both representative of a novel family of agonists of the extracellular calcium sensing receptor (CaSR) of possible clinical importance, are reported. The known chirality of the naphthylethylamine precursor has enabled assignment of the absolute configuration of both compounds, which is crucial for the receptor recognition. The crystal structures, though different, reveal for these agonists a notable absence of intramolecular pi-pi stacking between their respective aromatic groups. This suggests a common structural feature that allows CaSR agonists to be distinguished from antagonists, since in the latter, such interactions have been shown to be important. The connectivities between molecules in the crystal structures are also different, but both involve hydrogen bonding mediated by chloride ions as a common dominant feature. PMID- 19190387 TI - Three-dimensional networks in the 1:2 organic salts 2,2'-biimidazolium bis(3 carboxy-4-hydroxybenzenesulfonate) and 2,2'-bibenzimidazolium bis(3-carboxy-4 hydroxybenzenesulfonate) trihydrate. AB - The two title compounds of 2,2'-biimidazole (Bim) with 5-sulfosalicylic acid (5 H(2)SSA) and 2,2'-bibenzimidazole (Bbim) with 5-H(2)SSA are 1:2 organic salts, viz. C(6)H(8)N(4)(2+).2C(7)H(5)O(6)S(-), (I), and C(14)H(12)N(4)(2+).2C(7)H(5)O(6)S(-).3H(2)O, (II). The cation of compound (I) lies on a centre of inversion, whereas that of (II) lies on a twofold axis. Whilst compound (I) is anhydrous, three water molecules are incorporated into the crystal structure of (II). The substitution of imidazole H atoms by other chemical groups may favour the incorporation of water molecules into the crystal structure. In both compounds, the component cations and anions adopt a homogeneous arrangement, forming alternating cation and anion layers which run parallel to the (001) plane in (I) and to the (100) plane in (II). By a combination of N-H...O, O-H...O and C-H...O hydrogen bonds, the ions in both compounds are linked into three-dimensional networks. In addition, pi-pi interactions are observed between symmetry-related benzene rings of Bbim(2+) cations in (II). PMID- 19190388 TI - Two modes of O--H...O hydrogen bonding utilized in dimorphs of racemic 6-O acryloyl-2-O-benzoyl-myo-inositol 1,3,5-orthoformate. AB - The title compound, C(17)H(16)O(8), yields conformational dimorphs [forms (I) and (II)] at room temperature, separately or concomitantly, depending on the solvent of crystallization. The yield of crystals of form (I) is always much more than that of crystals of form (II). The molecule has one donor -OH group that can make intermolecular O-H...O hydrogen bonds with one of the two acceptor C=O groups, as well as with the hydroxyl O atom; interestingly, each of the options is utilized separately in the dimorphs. The crystal structure of form (I) contains one molecule in the asymmetric unit and is organized as a planar sheet of centrosymmetric dimers via O-H...O hydrogen bonds involving the OH group and the carbonyl O atom of the acryloyl group. In the crystal structure of form (II), which contains two independent molecules in the asymmetric unit, two different O H...O hydrogen bonds, viz. hydroxyl-hydroxyl and hydroxyl-carbonyl (benzoyl), connect the molecules in a layered arrangement. Another notable feature is the transformation of form (II) to form (I) via melt crystallization upon heating to 411 K. The higher yield of form (I) during crystallization and the thermal transition of form (II) to form (I) suggest that the association in form (I) is more highly favoured than that in form (II), which is valuable in understanding the priorities of molecular aggregation during nucleation of various polymorphs. PMID- 19190389 TI - Salts of maleic and fumaric acids with oxine: the role of isomeric acids in hydrogen-bonding patterns. AB - Both maleic and fumaric acid readily form adducts or complexes with other organic molecules. The 1:1 adduct formed by quinolin-8-ol (oxine) with maleic and fumaric acid are salts, namely 8-hydroxyquinolinium hydrogen maleate, C(9)H(8)NO(+).C(4)H(3)O(4)(-), (I), and 8-hydroxyquinolinium hydrogen fumarate, C(9)H(8)NO(+).C(4)H(3)O(4)(-), (II). The cations and anions of both salts are linked by ionic N(+)-H...O(-) hydrogen bonds. The maleate salt crystallizes in the space group P2(1)2(1)2(1), while the fumarate salt crystallizes in P?overline{1}. The maleic and fumaric acids in their complex forms exist as semimaleate and semifumarate ions (mono-ionized state), respectively. Classical N H...O and O-H...O hydrogen bonds, together with short C-H...O contacts, generate an extensive hydrogen-bonding network. The crystal structures of the maleate and fumarate salts of oxine have been elucidated to study the importance of noncovalent interactions in the aggregation and interaction patterns of biological molecules. The structures of the salts of the Z and E isomers of butenedioic acid (maleic and fumaric acid, respectively) with quinolin-8-ol are compared. PMID- 19190390 TI - Bis[2,4-diamino-5-(3,4,5-trimethoxybenzyl)pyrimidin-1-ium] DL-malate. AB - Racemic malic acid and trimethoprim [5-(3,4,5-trimethoxybenzyl)pyrimidine-2,4 diamine] form a 1:2 salt (monoclinic, P2(1)/c), 2C(14)H(19)N(4)O(3)(+).C(4)H(4)O(5)(2-), in which the malate component is disordered across a centre of inversion. The crystal structure of the salt consists of protonated trimethoprim residues and a malate dianion. The carboxylate group of the malate ion interacts with the trimethoprim cation in a linear fashion through pairs of N-H...O hydrogen bonds to form a cyclic hydrogen bonded motif. This is similar to the carboxylate-trimethoprim cation interaction observed earlier in the complex of dihydrofolate reductase with trimethoprim. The structure of the salt of trimethoprim with racemic DL-malic acid reported here is the first of its kind. The present study investigates the conformations and the hydrogen-bonding interactions, which are very important for biological functions. The pyrimidine plane makes a dihedral angle of 78.08 (7) degrees with the benzene ring of the trimethoprim cation. The cyclic hydrogen-bonded motif observed in this structure is self-organized, leading to novel types of hydrogen-bonding motifs in supramolecular patterns. PMID- 19190391 TI - Quinolin-6-ol at 100 K. AB - The title compound, C(9)H(7)NO, has two symmetry-independent molecules in the asymmetric unit, which have different conformations of the hydroxy group with respect to the quinoline ring. One of the molecules adopts a cis conformation, while the other shows a trans conformation. Each type of independent molecule links into a separate infinite O-H...N hydrogen-bonded chain with the graph-set notation C(7). These chains are perpendicular in the unit cell, one extended in the a-axis direction and the other in the b-axis direction. There is also a weak C-H...O hydrogen bond with graph-set notation D(2), which runs in the c-axis direction and joins the two separate O-H...N chains. The significance of this study lies in the comparison drawn between the experimental and calculated data of the crystal structure of the title compound and the data of several other derivatives possessing the hydroxy group or the quinoline ring. The correlation between the IR spectrum of this compound and the hydrogen-bond energy is also discussed. PMID- 19190392 TI - The influence of sulfur substituents on the molecular geometry and packing of thio derivatives of N-methylphenobarbital. AB - The room-temperature crystal structures of four new thio derivatives of N methylphenobarbital [systematic name: 5-ethyl-1-methyl-5-phenylpyrimidine 2,4,6(1H,3H,5H)-trione], C(13)H(14)N(2)O(3), are compared with the structure of the parent compound. The sulfur substituents in N-methyl-2-thiophenobarbital [5 ethyl-1-methyl-5-phenyl-2-thioxo-1,2-dihydropyrimidine-4,6(3H,5H)-dione], C(13)H(14)N(2)O(2)S, N-methyl-4-thiophenobarbital [5-ethyl-1-methyl-5-phenyl-4 thioxo-3,4-dihydropyrimidine-2,6(1H,5H)-dione], C(13)H(14)N(2)O(2)S, and N-methyl 2,4,6-trithiophenobarbital [5-ethyl-1-methyl-5-phenylpyrimidine-2,4,6(1H,3H,5H) trithione], C(13)H(14)N(2)S(3), preserve the heterocyclic ring puckering observed for N-methylphenobarbital (a half-chair conformation), whereas in N-methyl-2,4 dithiophenobarbital [5-ethyl-1-methyl-5-phenyl-2,4-dithioxo-1,2,3,4 tetrahydropyrimidine-6(5H)-one], C(13)H(14)N(2)OS(2), significant flattening of the ring was detected. The number and positions of the sulfur substituents influence the packing and hydrogen-bonding patterns of the derivatives. In the cases of the 2-thio, 4-thio and 2,4,6-trithio derivatives, there is a preference for the formation of a ring motif of the R(2)(2)(8) type, which is also a characteristic of N-methylphenobarbital, whereas a C(6) chain forms in the 2,4 dithio derivative. The preferences for hydrogen-bond formation, which follow the sequence of acceptor position 4 > 2 > 6, confirm the differences in the nucleophilic properties of the C atoms of the heterocyclic ring and are consistent with the course of N-methylphenobarbital thionation reactions. PMID- 19190393 TI - 4,6-Dinitro-N,N'-di-n-octylbenzene-1,3-diamine, 4,6-dinitro-N,N'-di-n undecylbenzene-1,3-diamine and N,N'-bis(2,4-dinitrophenyl)octane-1,8-diamine. AB - 4,6-Dinitro-N,N'-di-n-octylbenzene-1,3-diamine, C(22)H(38)N(4)O(4), (I), 4,6 dinitro-N,N'-di-n-undecylbenzene-1,3-diamine, C(28)H(50)N(4)O(4), (II), and N,N' bis(2,4-dinitrophenyl)octane-1,8-diamine, C(20)H(24)N(6)O(8), (III), are the first synthetic meta-dinitroarenes functionalized with long-chain aliphatic amine groups to be structurally characterized. The intra- and intermolecular interactions in these model compounds provide information that can be used to help understand the physical properties of corresponding polymers with similar functionalities. Compounds (I) and (II) possess near-mirror symmetry, with the octyl and undecyl chains adopting fully extended anti conformations in the same direction with respect to the ring. Compound (III) rests on a center of inversion that occupies the mid-point of the central C-C bond of the octyl chain. The middle six C atoms of the chain form an anti arrangement, while the remaining two C atoms take hard turns almost perpendicular to the rest of the chain. All three molecules display intramolecular N-H...O hydrogen bonds between the amine and nitro groups, with the same NH group forming a bifurcated intermolecular hydrogen bond to the nitro O atom of an adjacent molecule. In each case, these interactions link the molecules into one-dimensional molecular chains. In (I) and (II), these chains pack so that the pendant alkyl groups are interleaved parallel to one another, maximizing nonbonded C-H contacts. In (III), the alkyl groups are more isolated within the molecular chains and the primary nonbonded contacts between the chains appear to involve the nitro groups not involved in the hydrogen bonding. PMID- 19190394 TI - Renal function in the elderly and drug dosing. PMID- 19190395 TI - Reference values for CD4+ and CD8+ T lymphocytes with naive or memory phenotype and their association with mortality in the elderly. AB - BACKGROUND: Well-established reference values which take into account the influence of age on immune cell phenotype, and the impact of naive or memory T cells on mortality have not been well defined in the elderly. OBJECTIVE: The aim of this study was to evaluate the reference values for the peripheral number of total and naive or memory CD4 and CD8 T cells in a healthy population in Italy, and to analyze whether the immune phenotype was associated with an increased risk of death among older adults. METHODS: The number of total or naive and memory CD4+ or CD8+ T cells was evaluated in the peripheral blood of 288 healthy people ranging in age from 20 to 107 years. Furthermore, to correlate peripheral immune phenotype with mortality rate after a 3-years follow-up, a retrospective analysis was performed on the results from those individuals aged >65 years at the time of the enrollment in the study. RESULTS: The absolute number of total and naive T cells was progressively reduced with increasing age in both the CD4+ and CD8+ T cell populations. The decrease was particularly evident for cells with naive phenotype, since CD4-naive and CD8-naive T cells respectively showed a 4- and a 2 to 3-fold reduction in 70- to >90-year-old subjects in comparison with young adults. The number of CD4 memory T cells significantly increased with age. No significant age-related change was observed in the number of CD8+ memory T cells. Of the 194 subjects included in the study of association of immune phenotype with mortality, 121 were alive and 73 deceased during the 3-year follow-up. The impact of immune parameters on survival demonstrated that only the absolute number of CD8 memory T cells, after adjustment for age, correlated with increased mortality (OR 1.007, 95% CI 1.002-1.012, p = 0.01). The correlation was significant in female but not in male subjects. CONCLUSION: We provide reference values for total and naive or memory CD4 and CD8 T cell populations, and demonstrate that the absolute number of CD8 memory T cells, after adjustment for age, correlates with increased mortality. PMID- 19190396 TI - Effective determination of dry weight by intradialytic bioimpedance analysis in hemodialysis. AB - BACKGROUND: The new dry weight (DW) assessment technique by intradialytic sequential bioimpedance analysis (BIA) was evaluated in this study. METHODS: Intradialytic changes in the ratio of resistance at baseline per each time point (R(0)/R(t)) determined by a total body single-frequency BIA were recorded in 19 hemodialysis patients. The bioimpedance DW (BIADW) could be drawn from a time point at which R(0)/R(t) was constant. RESULTS: After the first session where the previous DW was prescribed, the participants could be divided into 2 groups: the under BIADW and over BIADW groups. In the under BIADW group, the BIADW was achieved earlier than the previous DW. The intradialytic symptoms disappeared without changes in the home-monitored mean blood pressure and antihypertensive agents at 1-month follow-up. In the over BIADW group, the BIADW could be obtained in the following sessions and the prescribed DW could be gradually reduced. The home-monitored mean blood pressure was decreased and the numbers of antihypertensive drugs were reduced. CONCLUSION: Intradialytic single-frequency BIA assessment can be used as a tool to accurately determine the DW. PMID- 19190397 TI - Long-term use of low-molecular-weight heparin in hemodialysis patients: a 7-year experience. AB - BACKGROUND: Low-molecular-weight-heparin (LMWH) is not routinely used as anticoagulant in hemodialysis (HD). The ideal dose and the safety of long-term use are not known. METHODS: A prospective three-phase interventional study. Phase 1 involved dose titration, phase 2 safety and efficacy and phase 3 routine practice. RESULTS: During 7 years of the use of the LMWH enoxaparin (EN), 236 patients were treated with a total number of 60,987 HD sessions. The mean dose used during the titration phase was 0.43 +/- 0.16 mg/kg/session, which was subsequently reduced in phase 3 to 0.36 +/- 0.14 mg/kg/session. The long-term effects of EN on the platelet count and lipid profile were comparable to unfractionated heparin. CONCLUSION: The long-term use of LMWH (EN) with a reduced dose in HD is practical and safe. PMID- 19190398 TI - Very early initiation of renal replacement therapy in critically ill patients with acute kidney injury: benefits and risks. PMID- 19190400 TI - Correlation between pulse wave velocity and fluid distribution in hemodialysis patients. AB - BACKGROUND: In this study, we focused on whether volume overload plays a role in the development of arterial stiffness. METHODS: Seventy-three prevalent hemodialysis patients were enrolled in a cross-sectional study. Arterial stiffness was assessed by carotid-femoral pulse wave velocity (PWV), and volume overload was assessed by bioimpedance analysis. RESULTS: Patients were divided into a high PWV group and a low PWV group based on the median of PWV. Age, systolic blood pressure, pulse pressure (PP), extracellular water/total body water ratio (ECW/TBW), diabetic status (all p < 0.01), and history of cardiac events (p < 0.05) were significantly higher in the high PWV group (n = 37) than in the low PWV group. In the correlation analysis, PWV was positively associated with PP, systolic blood pressure, ECW/TBW, age, history of cardiac events, and diabetes (all p < 0.01). Multiple regression analysis showed that PWV was independently related to PP and ECW/TBW (p < 0.01). CONCLUSION: Volume overload plays an important role in the development of arterial stiffness in hemodialysis patients. PMID- 19190401 TI - Biological mechanism of antidepressant effect of omega-3 fatty acids: how does fish oil act as a 'mind-body interface'? AB - The unsatisfactory results of monoamine-based antidepressant therapy and the high occurrence of somatic symptoms and physical illness in patients with depression imply that the serotonin hypothesis is insufficient to approach the aetiology of depression. Depressive disorders with somatic presentation are the most common form of depression. Somatization, the bodily symptoms without organic explanation, is similar to cytokine-induced sickness behaviour. Based on recent evidence, omega-3 polyunsaturated fatty acids (n-3 PUFAs, or n-3 fatty acids) are enlightening a promising path to discover the unsolved of depression, sickness behaviour and to link the connection of mind and body. The PUFAs are classified into n-3 (or omega-3) and n-6 (or omega-6) groups. Eicosapentaenoic acid and docosahexaenoic acid, the major bioactive components of n-3 PUFAs, are not efficiently synthesized in humans and should therefore be obtained directly from the diet, particularly by consuming fish. Docosahexaenoic acid deficiency is associated with dysfunctions of neuronal membrane stability and transmission of serotonin, norepinephrine and dopamine, which might connect to the aetiology of mood and cognitive dysfunction of depression. Likewise, eicosapentaenoic acid is important in balancing the immune function and physical health by reducing membrane arachidonic acid (an n-6 PUFA) and prostaglandin E(2) synthesis, which might be linked to the somatic manifestations and physical comorbidity in depression. The role of n-3 PUFAs in immunity and mood function supports the promising hypothesis of psychoneuroimmunology of depression and provides an excellent interface between 'mind' and 'body'. This review is to provide an overview of the evidence about the role of n-3 PUFAs in depression and its common comorbid physical conditions and to propose mechanisms by which they may modulate molecular and cellular functions. PMID- 19190402 TI - Inhella inkyongensis gen. nov., sp. nov., a new freshwater bacterium in the order Burkholderiales. AB - A freshwater bacterium, designated IMCC1713(T), was isolated from a highly eutrophic artificial pond. Cells of the strain were Gram-negative, chemoheterotrophic, polybeat and obligately aerobic short rods that were motile with a single polar flagellum. The 16S rRNA gene sequence similarity analysis showed that the novel strain was most closely related to the species Roseateles depolymerans (96.3%), Mitsuaria chitosanitabida (96.2%), Ideonella dechloratans (96.2%), and Pelomonas saccharophila (96.1%) in the Sphaerotilus-Leptothrix group within the order Burkholderiales. Phylogenetic trees based on 16S rRNA gene sequences indicated that the isolate formed an independent monophyletic clade within the order Burkholderiales. The relatively low DNA G+C content (57.4 mol%), together with several phenotypic characteristics, differentiated the novel strain from other members of the Sphaerotilus-Leptothrix group. From the taxonomic data, therefore, the strain should be classified as a novel genus and species, for which the name Inhella inkyongensis gen. nov., sp. nov. is proposed. The type strain of the proposed species is strain IMCC1713(T) (=KCTC 12791(T)=NBRC 103252(T)=CCUG 54308(T)). PMID- 19190403 TI - Functional analysis of MCNA, a gene encoding a catalytic subunit of calcineurin, in the rice blast fungus magnaporthe oryzae. AB - Magnaporthe oryzae, the causal agent of rice blast, forms a specialized infection structure, called an appressorium, which is crucial for penetration and infection of the host plant. Pharmacological data suggest that calcium/calmodulin-dependent signaling is involved in appressorium formation in this fungus. To understand the role of the calcium/ calmodulin-activated protein phosphatase on appressorium formation at the molecular level, MCNA, a gene encoding the catalytic subunit of calcineurin, was functionally characterized in M. oryzae. Transformants expressing sense/antisense RNA of MCNA exhibited significant reductions in mycelial growth, conidiation, appressorium formation, and pathogenicity. cDNA of MCNA functionally complemented a calcineurin disruptant strain (cmp1::LEU2 cmp2::HIS3) of Saccharomyces cerevisiae. These data suggest that calcineurin A plays important roles in signal transduction pathways involved in the infection related morphogenesis and pathogenicity of M. oryzae. PMID- 19190404 TI - Thiosulfate Oxidation and mixotrophic growth of Methylobacterium goesingense and Methylobacterium fujisawaense. AB - The mixotrophic growth with methanol plus thiosulfate was examined in nutrient limited mixotrophic condition for Methylobacterium goesingense CBMB5 and Methylobacterium fujisawaense CBMB37. Thiosulfate oxidation increased the growth and protein yield in mixotrophic medium that contained 150 mM methanol and 20 mM sodium thiosulfate, at 144 h. Respirometric study revealed that thiosulfate was the most preferable reduced inorganic sulfur source, followed by sulfite and sulfur. M. goesingense CBMB5 and M. fujisawaense CBMB37 oxidized thiosulfate directly to sulfate, and intermediate products of thiosulfate oxidation such as polythionates, sulfite, and sulfur were not detected in spent medium and they did not yield positive amplification for tested soxB primers. Enzymes of thiosulfate oxidation such as rhodanese and sulfite oxidase activities were detected in cell free extracts of M. goesingense CBMB5, and M. fujisawaense CBMB37, and thiosulfate oxidase (tetrathionate synthase) activity was not observed. It indicated that both the organisms use the "non-S4 intermediate" sulfur oxidation pathway for thiosulfate oxidation. It is concluded from this study that M. goesingense CBMB5, and M. fujisawaense CBMB37 exhibited mixotrophic metabolism in medium containing methanol plus thiosulfate and that thiosulfate oxidation and the presence of a "Paracoccus sulfur oxidation" (PSO) pathway in methylotrophic bacteria are species dependant. PMID- 19190405 TI - Investigation of the central carbon metabolism of Sorangium cellulosum: metabolic network reconstruction and quantification of pathway fluxes. AB - In the present work, the metabolic network of primary metabolism of the slow growing myxobacterium Sorangium cellulosum was reconstructed from the annotated genome sequence of the type strain So ce56. During growth on glucose as the carbon source and asparagine as the nitrogen source, So ce56 showed a very low growth rate of 0.23 d-(1), equivalent to a doubling time of 3 days. Based on a complete stoichiometric and isotopomer model of the central metabolism, 13C metabolic flux analysis was carried out for growth with glucose as carbon and asparagine as nitrogen sources. Normalized to the uptake flux for glucose (100%), cells recruited glycolysis (51%) and the pentose phosphate pathway (48%) as major catabolic pathways. The Entner-Doudoroff pathway and glyoxylate shunt were not active. A high flux through the TCA cycle (118%) enabled a strong formation of ATP, but cells revealed a rather low yield for biomass. Inspection of fluxes linked to energy metabolism revealed that S. cellulosum utilized only 10% of the ATP formed for growth, whereas 90% is required for maintenance. This explains the apparent discrepancy between the relatively low biomass yield and the high flux through the energy-delivering TCA cycle. The total flux of NADPH supply (216%) was higher than the demand for anabolism (156%), indicating additional reactions for balancing of NADPH. The cells further exhibited a highly active metabolic cycle, interconverting C3 and C4 metabolites of glycolysis and the TCA cycle. The present work provides the first insight into fluxes of the primary metabolism of myxobacteria, especially for future investigation on the supply of cofactors, building blocks, and energy in myxobacteria, producing natural compounds of biotechnological interest. PMID- 19190406 TI - Energy generation coupled to azoreduction by membranous vesicles from Shewanella decolorationis S12. AB - Previous studies have demonstrated that Shewanella decolorationis S12 can grow on the azo compound amaranth as the sole electron acceptor. Thus, to explore the mechanism of energy generation in this metabolism, membranous vesicles (MVs) were prepared and the mechanism of energy generation investigated. The membrane, which was fragmentized during preparation, automatically formed vesicles ranging from 37.5-112.5 nm in diameter under electron micrograph observation. Energy was conserved when coupling the azoreduction by the MVs of an azo compound or Fe(III) as the sole electron acceptor with H2, formate, or lactate as the electron donor. The amaranth reduction by the vesicles was found to be inhibited by specific respiratory inhibitors, including Cu(2+) ions, dicumarol, stigmatellin, and metyrapone, indicating that the azoreduction was indeed a respiration reaction. This finding was further confirmed by the fact that the ATP synthesis was repressed by the ATPase inhibitor N,N'-dicyclohexylcarbodiimide (DCCD). Therefore, this study offers solid evidence of a mechanism of microbial dissimilatory azoreduction on a subcell level. PMID- 19190408 TI - LC-MS/MS profiling-based secondary metabolite screening of Myxococcus xanthus. AB - Myxobacteria, Gram-negative soil bacteria, are a well-known producer of bioactive secondary metabolites. Therefore, this study presents a methodological approach for the high-throughput screening of secondary metabolites from 4 wild-type Myxococcus xanthus strains. First, electrospray ionization mass spectrometry (ESI MS) was performed using extracellular crude extracts. As a result, 22 metabolite peaks were detected, and the metabolite profiling was then conducted using the m/z value, retention time, and MS/MS fragmentation pattern analyses. Among the peaks, one unknown compound peak was identified as analogous to the myxalamid A, B, and C series. An analysis of the tandem mass spectrometric fragmentation patterns and HR-MS identified myxalamid K as a new compound derived from M. xanthus. In conclusion, LC-MS/MS-based chemical screening of diverse secondary metabolites would appear to be an effective approach for discovering unknown microbial secondary metabolites. PMID- 19190407 TI - Diverse Antibacterial activity of Pectobacterium carotovorum subsp.carotovorum isolated in Korea. AB - Fifty-four Pectobacterium carotovorum subsp. carotovorum strains isolated in Korea were characterized by a spectrum of antibacterial activities against 7 indicator strains chosen to represent various regions and host plants. All P. carotovorum subsp. carotovorum isolates tested could be grouped into 4 classes depending on the pattern of antibacterial substance production. All tested strains had DNA fragment(s) homologous to the genes encoding carotovoricin and 21 of them had genes homologous to DNA invertase. Sixteen strains had genes homologous to the genes encoding carocin S1. Several isolates produced antibacterial substances active against strains in Brenneria, Pantoea, and Pectobacterium genera that belonged formerly to the genus Erwinia. Strains in Pseudomonas or Xanthomonas sp. were not sensitive to the antibacterial substances produced by P. carotovorum subsp. carotovorum, except for X. albilineans that was sensitive to antibacterial substances produced by most strains in P. carotovorum subsp. carotovorum and P. betavasculorum KACC10056. These results demonstrated the diverse patterns of antibacterial substance production and the possibility of the existence of new antibacterial substance(s) produced by P. carotovorum subsp. carotovorum isolated in Korea. PMID- 19190409 TI - Characteristics of B-cell-specific growth substance produced by Bacillus licheniformis E1. AB - A B cell-specific growth substance (BGS) was isolated from the slime layer of Bacillus licheniformis E1. Unlike LPS, the BGS was not affected by polymixin B, an inhibitor of LPS, or by TLR4, and resulted in the growth of B cells. When BALB/c mice were treated with the BGS, the B cell population was found to increase in both the bone marrow and the spleen, with a marked increase after 24 h in the bone marrow and after 48 h in the spleen. When using antibodies to B cell lineage-restricted surface molecules to analyze the B cell population changes resulting from treatment with the BGS, an increase in immature B cells (IgM(+) and AA4.1(+)) and mature B cells (IgM(+) and IgD(+)) was found in the bone marrow 24 h after treatment with the BGS, whereas a decrease in mature B cells and increase in IgG(+) B cells were found in the spleen. When the BGS and OVA antigen were injected into the peritoneal cavity of BALB/c mice, this resulted in a high OVA-specific antibody titer in the sera, similar to that induced by aluminum hydroxide. Therefore, it is anticipated that the mass production of the BGS by B. licheniformis E1 could be used for studies of B cells in immunology, and contribute to the development of a new adjuvant for vaccine manufacture. PMID- 19190410 TI - A new strategy to improve the efficiency and sustainability of Candida parapsilosis catalyzing deracemization of (R,S)-1-phenyl-1,2-ethanediol under non growing conditions: increase of NADPH availability. AB - Microbial oxidoreductive systems have been widely used in asymmetric syntheses of optically active alcohols. However, when reused in multi-batch reaction, the catalytic efficiency and sustainability of non-growing cells usually decreased because of continuous consumption of required cofactors during the reaction process. A novel method for NADPH regeneration in cells was proposed by using pentose metabolism in microorganisms. Addition of D-xylose, L-arabinose, or D ribose to the reaction significantly improved the conversion efficiency of deracemization of racemic 1-phenyl-1,2-ethanediol to (S)-isomer by Candida parapsilosis cells already used once, which afforded the product with high optical purity over 97%e.e. in high yield over 85% under an increased substrate concentration of 15 g/l. Compared with reactions without xylose, xylose added to multi-batch reactions had no influence on the activity of the enzyme catalyzing the key step in deracemization, but performed a promoting effect on the recovery of the metabolic activity of the non-growing cells with its consumption in each batch. The detection of activities of xylose reductase and xylitol dehydrogenase from cell-free extract of C. parapsilosis made xylose metabolism feasible in cells, and the depression of the pentose phosphate pathway inhibitor to this reaction further indicated that xylose facilitated the NADPH-required deracemization through the pentose phosphate pathway in C. parapsilosis. moreover, by investigating the cofactor pool, the xylose addition in reaction batches giving more NADPH, compared with those without xylose, suggested that the higher catalytic efficiency and sustainability of C. parapsilosis non-growing cells had resulted from xylose metabolism recycling NADPH for the deracemization. PMID- 19190411 TI - Improving the productivity of recombinant protein in Escherichia coli under thermal stress by coexpressing GroELS chaperone system. AB - Here, we demonstrate that the overexpression of the GroELS chaperone system, which assists the folding of intracellular proteins and prevents aggregation of its biological targets, can enhance the thermotolerance of Escherichia coli strains and facilitate the production of recombinant protein under thermal stress. The overexpression of GroELS led to an about 2-fold higher growth rate of E. coli XL-1 blue than control at 45 degrees and induced the growth of the strain even at 50 degrees , although the growth was not sustained in the second-round culture. The effect of GroELS overexpression was also effective on other E. coli strains such as JM109, DH5alpha, and BL21. Finally, we have shown that coexpression of GroELS allows us to produce recombinant protein even at 50 degrees , a temperature at which the protein production based on E. coli is not efficient. This study indicates that the employment of the GroELS overexpression system can expand the range of environmental conditions for E. coli. PMID- 19190412 TI - Isolation of Hafnia species from kimchi. AB - Eight commercially packaged kimchi products were examined over 15 days of storage at 4 degrees to evaluate the occurrence of Hafnia alvei (H. alvei). Additionally, 7 saeujeot products, as a possible ingredient source, were analyzed to examine the bacteria's origin. Over the storage period, kimchi samples had decreasing pH levels, which stabilized at pH 4.2; acidity increased to 0.9+/-0.1%. Lactose nonfermenting bacteria, which H. alvei belongs to, gradually reduced in numbers over the kimchi storage. However, the relative frequency of H. alvei to lactose nonfermenting bacteria tended to increase. From the kimchi samples, 58 H. alvei presumptive colonies were selected. Forty-three colonies turned out to be H. alvei and 15 colonies were identified as other strains or of uncertain identification when the API 20E system was used. From further test, 3 of the 43 colonies were H. alvei (-) against the phage test. Finally, H. alvei was isolated from saeujeot, indicating that this ingredient can be an originating source of H. alvei in kimchi. PMID- 19190413 TI - One-step simultaneous immunochromatographic strip test for multianalysis of ochratoxin a and zearalenone. AB - Individual immunochromatographic assays (ICG) for ochratoxin A (OTA) and zearalenone (ZEA) were optimized and used in the development of a one-step simultaneous immunochromatographic assay (OS-ICG) for the rapid multianalysis of two mycotoxins in corn samples. The nitrocellulose membrane of the OS-ICG was treated with OTA-bovine serum albumin (BSA), ZEA-ovalbumin (OVA), and anti-mouse IgG in the OTA test, ZEA test, and control zones, respectively. Monoclonal antibody-gold conjugates (OTA3 MAb-gold and ZEA2C5 MAb-gold) were sprayed onto the conjugate pad. The visual detection limits were 2.5 and 5 ng/ml for OTA and ZEA, resepectively, and the results were obtained within 15 min after starting the analysis. An efficient, simple, and rapid extraction method using 30% MeOH/PBS was established and validated by analyzing the corn samples spiked with OTA/ZEA mixtures (0/0, 5/10, 10/20, and 20/30 microg/kg). The cut-off values of the OS-ICG for the spiked corn were 5 and 10 microg/kg for OTA and ZEA, respectively. Natural corn samples were analyzed by OS-ICG, direct competitive enzyme-linked immunosorbent assay (DC-ELISA), and HPLC. Results of the OS-ICG were in good agreement with those obtained by DC-ELISA and HPLC. The developed OS ICG offers a rapid, easy-to-use, and portable analytical system and can be used as a convenient qualitative tool for the on-site simultaneous determination of OTA and ZEA in cereals, food, and agricultural products in one analytical cycle. PMID- 19190414 TI - Aeration control of thermophilic aerobic digestion using fluorescence monitoring. AB - The thermophilic aerobic digestion (TAD) process is recognized as an effective method for rapid waste activated sludge (WAS) degradation and the deactivation of pathogenic microorganisms. Yet, high energy costs due to heating and aeration have limited the commercialization of economical TAD processes. Previous research on autothermal thermophilic aerobic digestion (ATAD) has already reduced the heating cost. However, only a few studies have focused on reducing the aeration cost. Therefore, this study applied a two-step aeration control strategy to a fill-and-draw mode semicontinuous TAD process. The NADH-dependent fluorescence was monitored throughout the TAD experiment, and the aeration rate shifted according to the fluorescence intensity. As a result, the simple two-step aeration control operation achieved a 20.3% reduction in the total aeration, while maintaining an effective and stable operation. It is also expected that more savings can be achieved with a further reduction of the lower aeration rate or multisegmentation of the aeration rate. PMID- 19190415 TI - Characterization of chryseobacterium aquaticum strain PUPC1 producing a novel antifungal protease from rice rhizosphere soil. AB - Strain PUPC1 produces an antifungal protease as well as plant growth promoting enzymes such as 1- aminocyclopropane-1-carboxylate (ACC) deaminase and phosphatase. Morphological, cultural, and physiological characteristics as well as 16S rRNA gene-sequence-based phylogenetic analysis confirmed the taxonomic affiliation of PUPC1 as Chryseobacterium aquaticum. The optimum growth of PUPC1 was observed at pH 6.0 and 30 degrees , and maximum protease production was observed in medium B amended with 1% tryptone, 0.5% sucrose, and 0.005% MnCl2. The protease was purified by ammonium sulfate precipitation, Sephadex G-75 gel filtration chromatography, and electroelution from preparative SDS-PAGE. The protease had a molecular mass of 18.5 kDa. The optimum pH and temperature stability of the protease were pH 5.0- 10.0 and temperature 40-70 degrees . Chryseobacterium aquaticum PUPC1 and its protease showed a broad-spectrum antifungal activity against phytopathogenic fungi. Strain PUPC1 also exhibited plant growth promoting traits. The objective of the present investigation was to isolate a strain for agricultural application for plant growth promotion and biocontrol of fungal diseases. PMID- 19190416 TI - Improved purification process for cholera toxin and its application to the quantification of residual toxin in cholera vaccines. AB - A simplified method for the purification of cholera toxin was developed. The 569B strain of Vibrio cholerae, a recognized hyper-producer of cholera toxin, was propagated in a bioreactor under conditions that promote the production of the toxin. The toxin was separated from the bacterial cells using 0.2-microm crossflow microfiltration, the clarified toxin was passed through the membrane into the permeate, and the bacterial cells were retained in the retentate. The 0.2-microm permeate was then concentrated 3-fold and diafiltered against 10 mM phosphate buffer, pH 7.6, using 30-kDa crossflow ultrafiltration. The concentrated toxin was loaded onto a cation exchange column, the toxin was bound to the column, and most of the impurities were passed unimpeded through the column. The toxin was eluted with a salt gradient of phosphate buffer, pH7.0, containing 1.0M NaCl. The peak containing the toxin was assayed for cholera toxin and protein and the purity was determined to be 92%. The toxin peak had a low endotoxin level of 3.1 EU/microg of toxin. The purified toxin was used to prepare antiserum against whole toxin, which was used in a G(M1) ganglioside-binding ELISA to determine residual levels of toxin in an oral inactivated whole-cell cholera vaccine. The G(M1) ganglioside-binding ELISA was shown to be very sensitive and capable of detecting as little as 1 ng/ml of cholera toxin. PMID- 19190417 TI - Planning a National Nursing Quality and Safety Alliance: strengthening nursing's policy voice. PMID- 19190418 TI - Code green: a new design imperative for healthcare facilities. PMID- 19190419 TI - Seasoned almost to perfection. PMID- 19190420 TI - Breaking news on social intelligence. PMID- 19190421 TI - The importance of positive culture in hospitals. PMID- 19190422 TI - Creating a positive work environment: implementation of the nurse-friendly hospital criteria. AB - The nursing shortage is negatively affecting the quality, effectiveness, and timeliness of patient care, which warrants a transformation of the nursing work environment. The Nurse-Friendly (NF) Hospital Project assisted 30 rural or small hospitals implement 12 NF criteria into hospital policies to create a positive work environment. The NF criteria represent organizational attributes necessary for ensuring quality patient care, positive work environment, professional development, and nurse satisfaction. The authors discuss nurses' perception of presence of NF criteria before and after they were incorporated at each hospital and the implementation outcomes. PMID- 19190423 TI - Clinical nurse leader impact on clinical microsystems outcomes. AB - The clinical nurse leader (CNL) role is being rapidly implemented in healthcare settings. A major component of the CNL role is to provide a leader at the center of the microsystem to promote quality outcomes. To examine the impact of CNLs at the Department of Veterans Affairs Tennessee Valley Healthcare System, outcomes from 5 diverse microsystems were evaluated before and after CNL implementation using electronic scheduling system reports, patient medical records, and quality improvement reports. Statistically significant improvements were found in all 5 of the indicators evaluated. The authors discuss the implications of improvement for clinical, satisfaction, and financial performance. PMID- 19190424 TI - Medication administration process assessment: applying lessons learned from commercial aviation. AB - Medication administration is a major safety issue for patients and providers. The authors describe a commercial aviation-based system safety assessment conducted on the medication administration process for a community teaching hospital in the northeast United States. Processes on 2 medical units and 1 surgical unit were assessed. A sampling of qualitative outcomes is presented in a risk prioritization framework, along with practical recommendations predicated on the valuable lessons learned in commercial aviation. PMID- 19190425 TI - Workplace bullying: concerns for nurse leaders. AB - OBJECTIVE: The aim of this study was to describe nurses' experiences with and characteristics related to workplace bullying. BACKGROUND: Although the concept of workplace bullying is gaining attention, few studies have examined workplace bullying among nurses. METHODS: This was a descriptive study using a convenience sample of 249 members of the Washington State Emergency Nurses Association. The Negative Acts Questionnaire-Revised was used to measure workplace bullying. RESULTS: Of the sample, 27.3% had experienced workplace bullying in the last 6 months. Most respondents who had been bullied stated that they were bullied by their managers/directors or charge nurses. Workplace bullying was significantly associated with intent to leave one's current job and nursing. CONCLUSION: In seeking remedies to the problem of workplace bullying, nurse leaders need to focus on why this bullying occurs and on ways to reduce its occurrence. This is a critical issue, since it is linked with nurse attrition. PMID- 19190426 TI - Organizational readiness for evidence-based practice. AB - OBJECTIVE: This study explored factors that affect the adoption or rejection of evidence-based practice (EBP) changes and differences in nurse manager and staff nurse perceptions about those factors. BACKGROUND: Roger's Diffusion of Innovations Theory explains relevant organizational strategies for guiding practice change. METHODS: The primary author developed the Evidence-Based Practice Changes Survey consisting of 12 items, completed by 92 nurses at a level 1 trauma center. RESULTS: Top barriers to EBP were insufficient time, lack of staff, and not having the right equipment and supplies. Top reasons to adopt EBP were having personal interest in the practice change, avoiding risk of negative consequences to the patient, and personally valuing the evidence. Several statistically significant differences emerged for demographic variables. CONCLUSION: Planning for EBP change must address barriers and facilitators to practice change and emphasize the benefit for patients and value of the practice change to nurses. PMID- 19190427 TI - Quantitative estimation of the recurrent laryngeal nerve irritation by employing spontaneous intraoperative electromyographic monitoring during anterior cervical discectomy and fusion. AB - STUDY DESIGN: Prospective, clinical study. OBJECTIVE: The objective of our study was to evaluate the role of laryngeal intraoperative electromyography (IEMG) in predicting the development of postoperative recurrent laryngeal nerve (RLN) palsy in patients undergoing anterior cervical discectomy and fusion (ACDF). We also attempted to develop a method to quantify the total IEMG irritation of the RLN. SUMMARY OF BACKGROUND DATA: RLN injury has been recognized as the most common ACDF-associated neurologic injury. It has been postulated, that the employment of laryngeal IEMG may identify the operative events leading to RLN injury and subsequent postoperative palsy. METHODS: Laryngeal IEMG monitoring was performed in 298 patients undergoing ACDF. Preexistent baseline EMG activity, amplitude, and duration of IEMG activity were recorded. The total amount of RLN irritation was expressed as an Irritation Score (IS) applying a specially designed mathematical equation incorporating the amplitude, the duration, and the presence of any baseline EMG irritation. The relationship of IEMG activity with parameters such as the number of operative levels, the duration of the procedure, the presence of any previous neck surgeries, and the type of the used retractor was examined. RESULTS: IEMG activity was recorded in 14.4% of our patients. Postoperative RLN injury occurred in 2.3% of our patients. The sensitivity of IEMG was 100%, the specificity 87%, the positive predictive value 16%, and its negative predictive value 97%. The calculated IS ranged between 0.28 and 3.47 (mean IS: 2.09). Significantly increased IEMG activity was found in patients with previous surgical intervention, patients undergoing multilevel procedures, long lasting procedures, and cases in which self-retained retractors were used. Likewise, significantly increased IS were observed in patients with previous surgeries and in cases where self-retained retractors were used. CONCLUSIONS: Our study indicates that laryngeal IEMG is a high-sensitivity modality that can provide real-time information and can potentially minimize the risk of operative RLN injury. PMID- 19190428 TI - Dynamic evaluation of the spinal cord in patients with cervical spondylotic myelopathy using a kinematic magnetic resonance imaging technique. AB - STUDY DESIGN: This prospective study analyzed preoperative and postoperative dynamic changes of the spinal cord in patients with cervical spondylotic myelopathy. OBJECTIVE: (1) To study preoperative kinematic characteristics of the spinal cord in patients with cervical spondylotic myelopathy and compare results with healthy individuals, (2) to understand the dynamic changes of the spinal cord after posterior decompression, and (3) to discover whether the degree of posterior shifting is correlated with surgical outcomes using kinematic magnetic resonance imaging (MRI). SUMMARY OF BACKGROUND DATA: Laminoplasty for cervical myelopathy increases the space occupied by the spinal cord leading to a decompressive effect on the cord. However, no consecutive studies have reported the kinematic characteristics of the cervical spine in patients with cervical spondylotic myelopathy both preoperatively and postoperatively. Additionally, there have been no reports investigating the effects of posterior cord shifting in the neutral and maximum flexion and extension positions on surgical outcomes after cervical laminoplasty. METHODS: Twenty cervical spondylotic myelopathy patients who underwent extensive laminoplasty and 20 healthy individuals were examined. Preoperative and postoperative MRI records were available in all cases. The cervical spines of the subjects were examined in the neutral and maximum flexion and extension positions using an MRI scanner. Sagittal T1-weighted images were obtained at 12 different angles. Images were analyzed for the distance between the dorsal edge of the vertebral column and the center of the cord at each disc level using NIH image software. RESULTS: Average cord distances (L value) in the neutral position and maximum extension position at C4/5 was significantly smaller than those at the other disc levels. The spinal cords of the patients after laminoplasty moved dorsally in the enlarged spinal canal in the neutral position, and in the maximum flexion and extension position. However, the degree of posterior spinal cord shifting was not correlated with surgical outcomes. CONCLUSIONS: Cord distances are relatively smaller at C4/5 and C5/6 levels, resulting in a narrowing of the posterior subarachnoid space with posterior cord compression in patients with cervical spondylotic myelopathy. The outcome of surgery was not correlated with the magnitude of postoperative backward shifting of the spinal cord, although the spinal cord of patients after posterior decompression moved significantly dorsally at any of the flexed, neutral, or extended spinal positions. Thus numerous factors might affect the postoperative outcomes. PMID- 19190429 TI - Long-term follow-up after open-window corpectomy in patients with advanced cervical spondylosis and/or ossification of the posterior longitudinal ligament. AB - STUDY DESIGN: Prospective study was performed in case with cervical spondylosis and ossification of the posterior longitudinal ligament (OPLL). OBJECTIVE: A novel anterior approach was defined and some patients were operated in this study. The late results of open-window corpectomy (OWC) were documented in a group of patients. SUMMARY OF BACKGROUND DATA: The aim of this study was to assess long-term clinical and radiologic outcomes after OWC in patients with advanced cervical spondylosis and/or OPLL. METHODS: Data were collected from 15 patients at 3 time points: preoperatively and at 15 months and 92 months postoperatively. The clinical parameters were pain (assessed by visual analog scale) and upper extremity function (assessed by Japanese Orthopedics Association) score. The radiologic parameters recorded from magnetic resonance imaging and computed tomography were lordosis angles (cervical and segmental) and cervical spinal canal diameter. Group means were calculated and findings at the 3 time points were statistically compared. Adjacent-segment degeneration was also evaluated at 92 months. RESULTS: Concerning visual analog scale and Japanese Orthopedics Association results, the 2 postoperative mean scores were both significantly better than the preoperative mean but were not statistically different. The postoperative mean results for cervical and segmental lordosis angles and spinal canal diameter were all significantly better than the respective preoperative means for these parameters. Comparisons of postoperative means revealed no significant change in cervical lordosis, segmental lordosis, or cervical spinal canal diameter from short-term to long-term follow-ups. Five patients showed mild degeneration at adjacent segments in long-term follow-up. CONCLUSIONS: Long-term follow-up indicates that OWC yields satisfactory clinical and radiologic outcomes in patients with anterior compressive cervical spondylosis and/or OPLL. OWC technique creates a more stable construct with 3 point fixation and offers better load sharing among implants and healthy vertebrae. Our observations suggest that OWC is a good surgical option for this patient group. PMID- 19190430 TI - The assessment of cervical foramina with oblique radiographs: the effect of film angle on foraminal area. AB - STUDY DESIGN: Radiographic evaluation of cadaveric cervical spine specimens. OBJECTIVE: Assess reliability and reproducibility of foraminal dimensions obtained from cervical radiographs of varying obliquity and determine optimal angles for visualizing foramina at each cervical spine level. SUMMARY OF BACKGROUND DATA: Oblique radiographs may be considered to assess cervical foraminal dimensions and are generally obtained 45 degrees from the anteroposterior (AP) orientation. Previous reports have suggested that foraminal area observed on these radiographs may be influenced by changes in obliquity so that certain film angles may be better suited for accurately assessing foramen size, depending on which level is being evaluated. METHODS: Radiographs of 4 human cadaveric cervical spines were obtained at 5-degree increments from 20 to 70 degrees relative to AP orientation, using both left and right sides of each specimen. Foraminal area was estimated by measuring height and width of each foramen and also with a freehand area measurement tool. Reliabilities of both methods were calculated. At each level, foraminal area was plotted against film angle and quadratic best-fit curves were used to determine the maximum area observed and the optimal angle of obliquity for assessment. RESULTS: Assessment of foraminal area using the height and width values was associated with good interobserver reliability, whereas the freehand method exhibited excellent reliability. The optimal film angles for calculating foraminal area increased from 46.3 degrees for C2-C3 to 56.1 degrees for C7-T1. The ideal film angle that minimized the overall error of measurement across the entire cervical spine was estimated to be 52.4 degrees. CONCLUSIONS: Optimal angles for visualizing lower cervical foramina are larger than those for upper cervical spine. To minimize overall loss of foraminal area throughout the entire cervical spine, oblique cervical radiographs should be obtained at an angle of approximately 52 degrees from the AP orientation. PMID- 19190431 TI - Current trends in spinal arthroplasty: an assessment of surgeon practices and attitudes regarding cervical and lumbar disk replacement. AB - STUDY DESIGN: Survey study involving orthopedic and neurosurgical spine surgeons. OBJECTIVE: To report the current practices and opinions of spine surgeons regarding cervical and lumbar total disk arthroplasty (TDA) as alternatives to arthrodesis for the treatment of degenerative conditions of the spine. SUMMARY OF BACKGROUND DATA: TDA represents an alternative surgical treatment for degenerative spinal conditions that may avoid the deleterious effects associated with fusion. Unfortunately, the prevailing opinions of surgeons about TDA have not been well characterized and the patterns of its utilization have not been documented previously in the literature. METHODS: A questionnaire was developed to record the attitudes of spine surgeons regarding cervical and lumbar TDA and to assess their utilization of this technology. This survey was distributed to all of the surgeons attending the 2007 "Contemporary Update on Disorders of the Spine" meeting in Whistler, British Columbia. RESULTS: One hundred thirteen of the 133 surgeons present at the meeting completed the questionnaire, corresponding to a return rate of 85%. The percentage of surgeons who had performed lumbar TDA was significantly higher than that for cervical TDA (42% vs. 30%, P=0.05). However, 81% of respondents stated that they were more likely to perform cervical TDA now compared with 1 year ago, whereas 64% indicated that they were less likely to perform lumbar TDA. The most frequently cited reasons for not performing both cervical and lumbar TDA were questions concerning long term outcomes and perceived difficulties with obtaining financial compensation from insurance companies; in addition, surgeons were also concerned about revising lumbar TDA cases. CONCLUSIONS: Although the results of this study confirm that the enthusiasm for TDA was shared by many of these respondents, it is clear that additional long-term, prospective, comparative data are required before this technology may be considered as a replacement for more traditional spinal fusion procedures. PMID- 19190432 TI - Direct lateral lumbar interbody fusion for degenerative conditions: early complication profile. AB - STUDY DESIGN/SETTING: A community hospital prospective, nonrandomized chart review. March 2004 to December 2006, 58 patients were treated. Adverse events: new complaints and increasing length of stay limit early mobilization and require consultation with other physicians or reoperation. These formed the focus of the study. OBJECTIVE: Assess patient demographics and adverse events related to direct lateral lumbar approach. SUMMARY OF BACKGROUND DATA: Clinicians advocate anterior column support for lumbar conditions. Minimally invasive stabilization of lumbar spine via direct lateral approach gained popularity owing to perceived decrease in patient morbidity. METHODS: Mild or major adverse events during hospital stay or within 6 weeks of discharge were considered early complications and designated as medical, approach, or implant related. A historical cohort of open posterior spinal fusion patients was used for comparison. RESULTS: Forty three female and 15 male patients with a mean age of 61 years formed the study group. Surgery performed included 38 single level, 19 2-level, and 1 3-level case. Adverse events occurred in 13 patients (22.4%); 8 events were approach, 3 medical, and 1 implant bone interface related. Major complications occurred in 5 patients (8.6%). Two patients (3.4%) with L4 nerve injury showed residual motor effects, at 1 year postoperatively. Significant differences were noted between single and 2-level cases; Estimated blood loss (EBL) and operative time. Open posterior fusion patients experienced greater operative time and increased EBL compared with minimally invasive cases. CONCLUSIONS: Major adverse events approximated 8.6% with approach-related complaints of nerve irritation nearing 3.4%. Mild complications occurred in 13.7% of patients. Meralgia paresthetica was a primary approach-related complaint. Most complaints significantly reduced by first postoperative visit. One patient (1.7%) had symptoms lasting over a year that did not adversely affect function. Significant finding related to exposure, that is, 1-versus 2-level cases. Overall morbidity reduction noted by EBL is considerably less compared with the historical cohort. Direct lateral lumbar interbody fusion has proven to be of value. PMID- 19190433 TI - Monosegmental transpedicular fixation for selected patients with thoracolumbar burst fractures. AB - STUDY DESIGN: A prospective cohort study on selected consecutive patients. OBJECTIVE: To evaluate the efficacy of an innovative operative technique called monosegmental transpedicular fixation for the treatment of some thoracolumbar burst fractures. SUMMARY OF BACKGROUND DATA: Short-segment pedicle screw instrumentation is accepted by many spinal surgeons as an acceptable technique for the treatment of thoracolumbar burst fractures. Preoperative evaluation using the spinal load-sharing makes this technique more reliable. To preserve more motion segments, some authors have advocated using monosegmental pedicle screw instrumentation (MSPI) to treat thoracolumbar fractures. However, up until now this kind of maneuver is only performed in cases of flexion distraction injuries. METHODS: A cohort of 20 patients with thoracolumbar burst fractures fulfilling the inclusion criteria were prospectively submitted to surgical treatment of monosegmental transpedicular fixation plus posterior fusion. All instrumentations were performed with pedicle screws inserted bilaterally into the fractured level and 1 adjacent level, either superior or inferior depending on the locating side of the intact endplate. All patients were followed up. The preoperative radiographs, the postoperative radiographs within 1 week of operation, and the radiographs of the most recent follow-up were evaluated for kyphosis correction recorded in the Sagittal Index and Load-Sharing Classification (LSC) index. The postoperative functional outcomes were evaluated using the Frankel Performance Scale together with the Denis Pain Scale. RESULTS: Eighteen patients were followed up successfully with an average final follow-up of 24.7+/-8.0 months. The focal kyphotic angulations were corrected satisfactorily with the mean Sagittal Index of preoperative 16.5+/-6.6 degrees, initial postoperative 4.0+/ 2.4 degrees, and latest follow-up 4.8+/-4.0 degrees. No obvious loss of correction occurred except for 2 patients who both scored 8 points on the LSC Score. Postoperatively, most patients attained both functional neurologic improvement and pain relief, and only a few complications were noted. CONCLUSIONS: For selected thoracolumbar burst fractures, MSPI can provide the same or better fixation and preserve more motion segments than other methods of posterior pedicle instrumentation. With preoperative evaluation using the spinal LSC system, MSPI is effective and reliable for the treatment of thoracolumbar burst fractures when properly indicated. PMID- 19190434 TI - Motion generated in the unstable lumbar spine during hospital bed transfers. AB - STUDY DESIGN: A parallel group design with repeated measures using a cadaver model was employed. OBJECTIVE: The purpose of this study was to evaluate and compare lumbar spine motion generated in the presence of spinal instabilities during common hospital moves using different transfer techniques. SUMMARY OF BACKGROUND DATA: Up to 25% of spinal cord injuries may occur during initial management of the patient with a compromised spinal column, when multiple transfers between diagnostic locations and operating, recovery and hospital rooms are often required. Few studies have compared methods of moving patients with lumbar spinal column injuries in hospital settings. METHODS: A global instability was created in 3 cadavers at L1 and sensors were attached to T12 and L2. A 3 dimensional electromagnetic tracking system (Liberty, Polhemus Inc) was used to measure flexion, lateral bending, and axial rotation while moving a cadaver from one bed to another to compare 2 transfer techniques used in hospitals: manual transfer and the On3, a motorized lateral transfer device (Hill-Rom, Batesville, IN). RESULTS: Significant increases in lumbar angulations (P<0.05 for all motions) were produced by manual transfers as compared with the On3 device. Manual transfers completed with 6 trained personnel produced 153% more flexion, 92% more axial rotation, and 177% more lateral bending than moves performed by 2 people with the On3 transfer device. CONCLUSIONS: The On3 device reduced motion in the patient with an unstable lumbar spine during hospital bed transfers with fewer personnel. PMID- 19190435 TI - A comparison of the effectiveness of waxed paper and Gore-Tex on the minimally invasive epidural fibrosis model. AB - STUDY DESIGN: Experimental animal study. OBJECTIVE: The authors conducted a study to determine the effectiveness of waxed paper in preventing postlaminectomy epidural fibrosis in rats. SUMMARY OF BACKGROUND DATA: Extensive epidural fibrosis after lumbar surgery may be the underlying cause in most cases of failed back surgery syndrome. Various materials have been used to prevent epidural fibrosis, but only moderate success has been shown. METHODS: Laminectomies were performed at the fourth lumbar vertebra (L-4) in 30 rats. Waxed paper or Gore-Tex was applied in a blinded fashion to the operative sites. In the control group, only the L-4 laminectomy was performed. The rats were killed 28 days after surgery. RESULTS: The extent of epidural fibrosis was evaluated by histologic analysis. There was a meaningful statistical difference among the waxed paper group and the Gore-Tex group compared with the control group, but there was no difference when the effectiveness of waxed paper was compared with that of Gore Tex. CONCLUSIONS: In this experimental model, the waxed paper applied locally effectively reduced epidural fibrosis, completely avoided dural adherence, and induced no side effects. PMID- 19190436 TI - An evaluation of information on the Internet of a new device: the lumbar artificial disc replacement. AB - STUDY DESIGN: An analysis of websites, accessible to the public, was conducted pertaining to the lumbar artificial disc replacement. OBJECTIVE: The objective was to investigate the content of information available on the Internet pertaining to the lumbar artificial disc replacement. SUMMARY OF BACKGROUND DATA: The Internet is widely used by patients as an educational tool for health care information. Additionally, the Internet is used as a medium for direct-to consumer marketing. Recent approval of the lumbar artificial disc replacement has led to the emergence of numerous websites offering information about this procedure. It is thought that patients can be influenced by information found on the Internet; therefore, it is imperative that this information be accurate and as complete as possible. METHODS: Three commonly used search engines were used to locate 105 (35/search engine) websites providing information about the lumbar artificial disc replacement. Each website was evaluated with regard to authorship and content. RESULTS: Fifty-nine percent of the websites reviewed were authorized by a private physician group, 9% by an academic physician group, 6% by industry, 11% were news reports, and 15% were not otherwise categorized. Seventy-two percent offered a mechanism for direct contact and 30% provided clear patient selection criteria. Benefits were expressed in 87% of websites, whereas associated risks were described in 28% or less. European experiences were noted in 53%, whereas only 22% of websites detailed the current US experience. CONCLUSIONS: The results of this study demonstrate that much of the content of Internet-derived information pertaining to the lumbar artificial disc replacement is potentially misleading. Until long-term data are available, patients should be cautioned when using the Internet as a source for health care information, particularly with regard to the lumbar artificial disc replacement. PMID- 19190437 TI - Resolution of intraspinal retro-odontoid cyst associated with os odontoideum after posterior fixation. AB - STUDY DESIGN: A case report and review of previous literature. OBJECTIVES: To describe the diagnosis and successful treatment of an intraspinal retro-odontoid cyst associated with an os odontoideum. SUMMARY OF BACKGROUND DATA: Retro odontoid cysts associated with atlantoaxial instability are extremely rare. Direct excision of the cysts has commonly been performed for the surgical treatment of myelopathy. METHODS: A retro-odontoid cyst in a 58-year-old woman with os odontoideum was treated surgically by posterior fixation between C1 and C3 without resection of the retro-odontoid mass. The patient's history, clinical examination, imaging findings, and treatment are reported. RESULTS: The patient experienced walking difficulty, numbness in the upper and lower limbs, and hypesthesia in both hands. Congenital C2/3 fusion and atlantoaxial instability associated with an os odontoideum were found in the radiographs. Magnetic resonance imaging detected a round retro-odontoid mass, which compressed the spinal cord. The mass showed uniform low intensity on T1-weighted images and uniform high signal intensity on T2-weighted images. Gadolinium diethylenetriaminepenta-acetic acid-enhanced T1-weighted images showed rim enhancement of the mass with no internal enhancement. Three months after the operation, the mass was no longer visible in a magnetic resonance imaging examination and the patient completely recovered motor function in her extremities. CONCLUSIONS: Posterior fixation without resection was successfully used to eliminate a retro-odontoid cyst associated with atlantoaxial instability. PMID- 19190438 TI - Transvertebral herniotomy for T2/3 disc herniation--a case report. AB - STUDY DESIGN: A case report of a 51-year-old man with acute myelopathy owing to T2/3 disc herniation that was treated with transvertebral herniotomy. OBJECTIVES: To report surgical advantages of the transvertebral approach in the upper thoracic spine. SUMMARY OF BACKGROUND DATA: Various surgical approaches to the upper thoracic spine have been reported because the approach is difficult owing to the specific anatomical structure. However, a lack of consensus still remains regarding the choice of operative procedure because of some problems for each approach. METHODS: A 51-year-old man presented acute paraparesis of lower extremities and bladder paralysis owing to T2/3 disc herniation. The herniated disc was removed microscopically by the anterior approach through a 10-mm diameter hole made in the T2 vertebral body without sternum splitting. RESULTS: Satisfactory decompression was performed. After operation, the patient had full clinical motor and sensory recovery. CONCLUSIONS: Transvertebral approach, which has been recently performed for cervical disc lesion, was also less invasive and safer than the conventional approaches, such as sternum splitting, transthoracic or posterolateral approaches, for our patient with T2/3 disc herniation. PMID- 19190439 TI - Surgical management of thoracic spinal cord herniation: technical consideration. AB - STUDY DESIGN: This report describes a case of spinal cord herniation (SCH) and the surgical technique used to repair the herniation. OBJECTIVE: To describe a new surgical technique used to provide increased exposure to minimize spinal cord traction for safe repair of the ventral dural defect. SUMMARY OF BACKGROUND DATA: SCH is a relatively rare pathologic condition that is frequently misdiagnosed. It is characterized by the spontaneous herniation and tethering of the spinal cord through a ventral dural defect. A limitation of current surgical management is sufficient exposure of the defect through which the cord herniates. METHODS: This technique entails a posterior decompressive laminectomy and bilateral transpedicular approach, followed by unilateral removal of the pedicle and transverse processes to sufficiently expose the dural defect and SCH. A Gore-Tex graft is then used to repair the dural defect. RESULTS: This approach provided greater exposure of the ventral dural defect and anterior spinal cord, and allowed for safer dissection of dural adhesions. By releasing these adhesions, the spinal cord was returned to its normal position in the thecal sac. The dural defect was then repaired with minimal manipulation of the spinal cord. CONCLUSIONS: We present a rare case of SCH. We describe a new technique used to repair SCH, which adds to the repertoire of existing surgical techniques. By providing a wide exposure, this approach provides wide access to the ventral thecal sac, thus minimizing the requirement for cord traction and allowing for safer closure of the dural defect. PMID- 19190440 TI - Traumatic subacute spinal subdural hematoma successfully treated with lumbar drainage: case report. AB - STUDY DESIGN: A case report. OBJECTIVE: To report a rare case of traumatic subacute spinal subdural hematoma (SDH) successfully treated by lumbar drainage. SUMMARY OF BACKGROUND DATA: Few reports on traumatic spinal SDH are available in the English literature. METHODS: The authors present a rare case of traumatic subacute spinal SDH. A 43-year-old man developed severe pain in his back and both legs approximately 3 weeks after a trivial car accident that occurred 1 month before presentation. The patient reported paresthesia in the L5 and S1 dermatomes bilaterally and a walking difficulty because of severe leg pain. He assessed his pain severity as 8 using the visual analog pain score scale. Magnetic resonance imaging of the lumbar spine revealed a spinal SDH extending from the level of the L5 vertebral body to S1-S2 disc space. RESULTS: The spinal SDH was drained by lumbar puncture at the L5-S1 level without surgical exploration. The intensity of the pain decreased considerable immediately, and he was able to walk independently. The patient recovered uneventfully and a follow-up magnetic resonance imaging conducted 4 weeks after the procedure revealed complete disappearance of the hematoma. CONCLUSIONS: Traumatic spinal SDH is extremely rare and is usually associated with intracranial injury. Here, the authors report a rare case of traumatic spinal SDH not associated with head injury, which was successfully treated by lumbar drainage. PMID- 19190442 TI - Ectopic recurrence of craniopharyngiomas: case report. AB - OBJECTIVE: Craniopharyngiomas are benign tumors that originate from squamous cell rests of the embryonal hypophyseal-pharyngeal duct located along the pituitary stalk. After their surgical resection, recurrence usually occurs in the region of the original tumor bed. Ectopic recurrence of craniopharyngiomas is extremely rare. It usually occurs either along the surgical route, because of direct surgical seeding, or at a distal location in the subarachnoid space, because of seeding along the cerebrospinal fluid pathways. We present 3 examples of ectopic recurrences of craniopharyngiomas. CLINICAL PRESENTATION: The first patient was a 52-year-old woman with a history of resected suprasellar craniopharyngioma presenting 15 years later with a history of balance problems and new onset of double vision. Her magnetic resonance imaging scan revealed a tumor in the prepontine cistern. The second patient was a 41-year-old man with a history of a resected suprasellar craniopharyngioma presenting 9 years later with headache, dizziness, and disequilibrium. He was noted by his family to have an altered behavior with progressively increasing indifference. His magnetic resonance imaging scan showed a right frontal lesion in the vicinity of the sylvian fissure. The third patient was a 24-year-old man with a history of suprasellar craniopharyngioma resection, followed by conventional radiotherapy 12 years before his recent presentation with headache, numbness of the right side of his face, and increased drowsiness. His magnetic resonance imaging scan showed a bilateral cystic cerebellopontine angle lesion. INTERVENTION: The first patient underwent operation via a petrosal approach with subtotal resection of the tumor and decompression of the brainstem; this patient had an uneventful postoperative course. The tumor in the second patient was surgically resected through a pterional craniotomy, with an uneventful postoperative course. The third patient's right-sided cerebellopontine angle lesion was microsurgically resected, and the patient was given a single-dose gamma knife for the left-side and residual small right-side tumor. The histological diagnosis of all 3 lesions was craniopharyngioma. CONCLUSION: Although ectopic recurrence of a craniopharyngioma is very rare, it should always be considered in the differential diagnosis of what appears to be a new tumor in a patient with a history of previously resected craniopharyngiomas. Long-term follow-up of patients with resected craniopharyngioma is very important. PMID- 19190443 TI - Multiple primary cranial Ewing's sarcoma in adulthood: case report. AB - OBJECTIVE: Ewing's sarcoma is a malignant bone tumor occurring most frequently in the long bones and flat bones as a solitary lesion during the first 2 decades of life. Ewing's sarcoma and peripheral primitive neuroectodermal tumor have recently been considered to be the same entity because of histological and molecular similarities. CLINICAL PRESENTATION: A 25-year-old man presented with swelling in the right parietal region. Magnetic resonance imaging scans showed 2 mixed intense mass lesions in the right parietal and left frontal areas with heterogeneous gadolinium enhancement. A computed tomographic scan revealed osteolytic changes of the inner calvarial bone. Ga-67 citrate scintigraphy demonstrated abnormal uptake in the right parietal and the left frontal areas consistent with the findings on magnetic resonance imaging scans. There was no uptake lesion beyond the cranium. The patient underwent surgical excision of the right parietal lesion. INTERVENTION: Histological examinations showed densely packed, small round cells with rosette formation. The cells had scanty clear cytoplasm, and regular vesicular and hyperchromatic nuclei. Electron microscopy showed little differentiation to neuronal tissue, indicating Ewing's sarcoma. After surgical treatment, conventional whole cranial irradiation of 40 Gy and chemotherapy were conducted. The tumor in the left frontal region disappeared. Follow-up examinations showed no evidence of recurrence 1 year after the surgery. CONCLUSION: Although quite rare, Ewing's sarcoma should be taken into consideration as a differential diagnosis of multiple cranial mass lesions in adulthood. PMID- 19190444 TI - Intradural clival chordoma and ecchordosis physaliphora: a challenging differential diagnosis: case report. AB - OBJECTIVE: Purely intradural clival chordomas are rare neoplasms, and only a few cases have been reported. The reported cases present features similar to ecchordosis physaliphora, which is a notochordal remnant. We describe these 2 entities and their differential diagnoses, clinical courses, and management. This is the first reported case to be treated using a neuroendoscopic technique. CLINICAL PRESENTATION: A 60-year-old man presenting with memory loss underwent magnetic resonance imaging, which revealed an intradural retroclival mass without bone involvement. INTERVENTION: The patient underwent an endoscopic transsphenoidal-transclival procedure with subtotal removal of the tumor. Histological findings confirmed the diagnosis of a chordoma. CONCLUSION: Even if some parameters exist for a differential diagnosis, ecchordosis physaliphora and intradural chordoma may represent different aspects of the spectrum of the same pathology. Intradural clival chordomas have a better prognosis with respect to classic chordomas. Therefore, in subtotal removal such as that performed in our case, postoperative radiation therapy should be performed only if a regrowth of the remnant is seen during neuroradiological follow-up. PMID- 19190445 TI - Near-complete pathological response of chordoma to high-dose single-fraction radiotherapy: case report. AB - OBJECTIVE: High-dose, single-fraction radiotherapy, also known as radiosurgery, has theoretical advantages for the treatment of chordoma, which is considered resistant to conventional radiation therapy. Demonstration of biological effectiveness, as indicated by pathological response of a chordoma specimen after such radiotherapy, has not previously been reported. CLINICAL PRESENTATION: A 72 year-old man presented with lower back pain and was found to have an L3 vertebral mass. Open biopsy yielded chordoma. He was considered for en bloc resection, but definitive radiation therapy was preferred owing to medical comorbidities and concern for field contamination from the previous biopsy. INTERVENTION: The patient underwent single-fraction, high-dose, image-guided radiation therapy to the tumor. Two months later, he underwent kyphoplasty because of recurrent pain. A core biopsy performed at that time showed viable chordoma. Four months after treatment, he underwent L3 corpectomy because of symptoms of mechanical instability. Pathology now showed near-complete necrosis of the resected chordoma. CONCLUSION: Preoperative, single-fraction radiotherapy for chordoma induced a near-complete pathological response in this patient. That the effect was seen at 4 months but not 2 months after treatment suggests a time-dependent effect. This case suggests a promising role for single-fraction, image-guided radiation therapy in the treatment of chordoma. PMID- 19190446 TI - Delayed symptomatic coil migration after initially successful balloon-assisted aneurysm coiling: technical case report. AB - OBJECTIVE: To describe delayed migration of a coil loop after adjunctive balloon remodeling of an anterior communicating artery aneurysm. METHODS: A 56-year-old man with a subarachnoid hemorrhage from a small anterior communicating artery aneurysm underwent successful coil embolization with adjunctive balloon remodeling. RESULTS: Eight days after the procedure, the patient returned with the acute onset of right lower extremity paralysis. Magnetic resonance imaging demonstrated an acute left anterior cerebral artery territory infarction. Angiography demonstrated that this infarct was secondary to the delayed migration of a coil loop out of the aneurysm and into the left A1 to A2 junction. CONCLUSION: Delayed migration of a coil loop after adjunctive balloon remodeling represents a rare but potentially severe complication of this technique. PMID- 19190447 TI - Novel surgical treatment of a transverse-sigmoid sinus aneurysm presenting as pulsatile tinnitus: technical case report. AB - OBJECTIVE: Pulsatile tinnitus is a relatively common, potentially incapacitating condition that is often vascular in origin. We present a case of disabling pulsatile tinnitus caused by a transverse-sigmoid sinus aneurysm that was surgically treated with self-tying U-clips (Medtronic, Inc., Memphis, TN). We also review the literature and discuss other described interventions. CLINICAL PRESENTATION: A 48-year-old woman presented with a 5-year history of progressive pulsatile tinnitus involving the right ear. Her physical examination was consistent with a lesion that was venous in origin. Angiography demonstrated a wide-necked venous aneurysm of the transverse-sigmoid sinus that had eroded the mastoid bone. INTERVENTION: The patient underwent a retromastoid suboccipital craniectomy to expose the aneurysm and surrounding anatomy. The aneurysm dome was tamponaded and the aneurysm neck was coagulated until the dome had shrunk to a small remnant. The linear defect in the transverse sigmoid junction was then reconstructed with a series of U-clips and covered with Gelfoam hemostatic sponge (Pfizer, Inc., New York, NY). The patient awakened without neurological deficit and with immediate resolution of her tinnitus. A postoperative angiogram demonstrated obliteration of the aneurysm, with minimal stenosis in the region of the repair and good flow through the dominant right transverse-sigmoid junction. CONCLUSION: This technical case report describes a novel definitive surgical treatment of venous sinus aneurysms. This technique does not necessitate long term anticoagulation, has a low likelihood of reintervention, and provides immediate resolution of pulsatile tinnitus. PMID- 19190452 TI - Results, outcomes, and follow-up of remnants in the treatment of ophthalmic aneurysms: a 16-year experience of a combined neurosurgical and endovascular team. AB - OBJECTIVE: Ophthalmic aneurysms present unique challenges to a vascular team. This study reviews the 16-year experience of a multidisciplinary neurovascular service in the treatment, complications, outcomes, and follow-up of patients with ophthalmic aneurysms from 1990 to 2005. METHODS: A retrospective analysis of prospectively collected data of 134 patients with 157 ophthalmic aneurysms is presented. Subgroup analysis is performed based on treatment and clinical presentation of the patients. RESULTS: Clinical outcomes are reported using the Glasgow Outcome Scale. A "good" outcome is defined as a Glasgow Outcome Scale score of 4 or 5, and a "poor" outcome is defined as a Glasgow Outcome Scale score of 1 to 3. Outcome was related to patient age (P = 0.0002) and aneurysm size (P = 0.046). Outcomes for patients with ruptured aneurysms were related to hypertension (P < 0.0001) and clinical admission grade (P = 0.001). In patients with unruptured aneurysms, a good clinical outcome was noted in 103 (92.7%) of 111 patients at discharge and 83 (94.3%) of 88 patients at the time of the 1-year follow-up evaluation. Complete clipping was attained in 89 (79.5%) of 112 patients with angiographic follow-up. Patients with aneurysm remnants from both coiling and clipping had a low risk of regrowth, and there were no rehemorrhages. One of 25 patients with angiographic follow-up (average, 4.3 +/- 4.1 years) after "complete" clipping showed recurrence of the aneurysm. CONCLUSION: Despite the difficulties presented by ophthalmic aneurysms, these lesions can be successfully managed by a multidisciplinary team. Imaging follow-up of patients is important, as there is a risk of aneurysm regrowth after either coiling or clipping. PMID- 19190451 TI - High-intensity focused ultrasound surgery of the brain: part 1--A historical perspective with modern applications. AB - The field of magnetic resonance imaging-guided high-intensity focused ultrasound surgery (MRgFUS) is a rapidly evolving one, with many potential applications in neurosurgery. The first of 3 articles on MRgFUS, this article focuses on the historical development of the technology and its potential applications in modern neurosurgery. The evolution of MRgFUS has occurred in parallel with modern neurological surgery, and the 2 seemingly distinct disciplines share many of the same pioneering figures. Early studies on focused ultrasound treatment in the 1940s and 1950s demonstrated the ability to perform precise lesioning in the human brain, with a favorable risk-benefit profile. However, the need for a craniotomy, as well as the lack of sophisticated imaging technology, resulted in limited growth of high-intensity focused ultrasound for neurosurgery. More recently, technological advances have permitted the combination of high-intensity focused ultrasound along with magnetic resonance imaging guidance to provide an opportunity to effectively treat a variety of central nervous system disorders. Although challenges remain, high-intensity focused ultrasound-mediated neurosurgery may offer the ability to target and treat central nervous system conditions that were previously extremely difficult to address. The remaining 2 articles in this series will focus on the physical principles of modern MRgFUS as well as current and future avenues for investigation. PMID- 19190453 TI - Cranial dural arteriovenous fistulae: asymptomatic cortical venous drainage portends less aggressive clinical course. AB - OBJECTIVE: Cranial dural arteriovenous fistulae (dAVF) with cortical venous drainage (CVD) (Borden Types 2 and 3) are reported to carry a 15% annual risk of intracranial hemorrhage (ICH) or nonhemorrhagic neurological deficit (NHND). The purpose of this study was to compare the clinical course of Type 2 and 3 dAVFs that present with ICH or NHND with those that do not. METHODS: Twenty-eight patients with Type 2 or 3 dAVFs were retrospectively evaluated. CVD was classified as asymptomatic (aCVD) if patients presented incidentally or with pulsatile tinnitus or orbital phenomena. CVD was classified as symptomatic (sCVD) if patients presented with ICH or NHND. Occurrence of new ICH or new or worsening NHND between diagnosis and disconnection of CVD or last follow-up (if not disconnected) was noted. Overall frequency of events was compared using Fisher's exact test. Cumulative, event-free survival was compared using Kaplan-Meier analysis with log-rank testing. RESULTS: Of 17 patients with aCVD, 1 (5.9%) developed ICH and none experienced NHND or death during the median 31.4-month follow-up period. Of 11 patients with sCVD, 2 (18.2%) developed ICH and 3 (27.3%) experienced new or worsened NHND over the median 9.7-month follow-up period. One of these patients subsequently died. Overall frequency of ICH or NHND was significantly lower in patients with aCVD versus sCVD (P = 0.022). Respective annual event rates were 1.4 versus 19.0%. aCVD patients had significantly higher cumulative event-free survival (P = 0.0016). CONCLUSION: Cranial dAVFs with aCVD may have a less aggressive clinical course than those with sCVD. PMID- 19190454 TI - A national internet-based survey of cerebral aneurysm preference-based quality of life. AB - OBJECTIVE: Preference-based quality of life (QOL) instruments integrate all factors contributing to QOL and provide a comprehensive valuation of a health state. QOL values of the general public for cerebral aneurysm health states are not well understood. METHODS: In an internet-based survey of a nationally representative sample of the United States population, we measured standard gamble QOL values for the subjects' own health and for low-, medium-, and high risk cerebral aneurysm health state scenarios and determined the effect of depicting aneurysm-associated stroke and death risk as annual or cumulative. RESULTS: The 1654 subjects matched the United States population demographics. The subjects' mean (standard deviation) QOL for their current health was 0.82 (0.19), and for a cerebral aneurysm it was 0.78 (0.19) (P < 0.001). Mean low-, medium-, and high-risk aneurysm QOL values were 0.01, 0.06, and 0.13 lower than for current health, respectively (P < 0.001). The average discrepancy between aneurysm QOL and current health narrowed with age: 18 to 25 years, 0.09; 24 to 44 years, 0.06; 45 to 64 years, 0.03; and 65+ years, 0.01 (trend P < 0.001). Subjects who received only annual risk data provided the highest mean aneurysm QOL values (0.81 [0.18]); those who received both annual and 20-year cumulative risk information gave intermediate values (0.79 [0.18]), and those who received only data on cumulative 20-year risk provided the lowest values (0.76 [0.20]) (P < 0.001). CONCLUSION: Preference-based QOL values for cerebral aneurysms derived from the general public vary with the subjects' age, the risk of aneurysmal stroke and death, and the mathematical terminology used to convey the risk of stroke and death. PMID- 19190455 TI - Intraoperative direct third ventriculostomy and aqueductal stenting in deep seated midline brain tumor surgery. AB - OBJECTIVE: Surgical resection of deep-seated midline brain tumors does not always resolve obstruction of cerebrospinal fluid pathways, and an additional operation- ventricular shunting--is required. To prevent postoperative obstructive hydrocephalus, we combine tumor removal and internal ventricular shunting in 1 stage. METHODS: Between 2000 and 2006, 82 patients with deep-seated midline brain tumors (tumors of the third ventricle, pineal region, thalamus, upper brainstem, and superior half of the fourth ventricle) underwent 84 tumor resections with intraoperative internal ventricular shunting. Two types of intraoperative shunting were performed: direct third ventriculostomy with fenestration of the premammillary membrane of the third ventricle floor and Liliequist's membrane, 53 operations; and aqueductal stenting, 30 operations. In 1 patient, third ventriculostomy and aqueductal stenting were performed simultaneously. RESULTS: As most of the tumors had an infiltrative growth pattern, gross total tumor removal was achieved in only 31% of patients in this series. There were no fatal outcomes related to the surgery. Follow-up data were collected in 73 patients (89%) and ranged from 2 to 68 months (median, 16 months). Additional shunting because of inadequate function of stoma or stent was performed in 13 patients at various times after surgery (median, 30 days). The Kaplan-Meier survival analysis demonstrated that at 12 and 24 months the intraoperative direct third ventriculostomy success rates were 67 and 61%, respectively; aqueductal stenting success rates were 93% at both 12 and 24 months. CONCLUSION: Intraoperative direct third ventriculostomy and aqueductal stenting under direct visual control were found to be reliable methods of hydrocephalus management in patients with deep-seated midline brain tumors. PMID- 19190456 TI - Endoscopic endonasal approach for clival chordomas. AB - OBJECTIVE: Cranial base chordomas are difficult lesions to treat. The endoscopic endonasal approach (EEA) takes advantage of the natural sinus corridor and may provide a less invasive approach for these midline tumors. METHODS: Patients undergoing EEA for chordomas were selected from a database of more than 800 consecutive patients undergoing EEA at the University of Pittsburgh Medical Center and were retrospectively evaluated. Additionally, a systematic review of the literature of endoscopic endonasally resected chordomas was performed and compared with our personal experience. RESULTS: Twenty patients (8 females and 12 males) underwent 26 endoscopic EEAs for cranial base chordomas. Eight chordomas (40%) were recurrent. Treatment of the 12 newly diagnosed chordomas included 8 total resections (66.7%), 2 near total resections (16.7%), and 2 subtotal resections (16.7%). Treatment of the 8 recurrent chordomas included 1 gross total resection (12.5%), 2 near total resections (25.0%), and 5 subtotal resections (62.5%). Two patients (10%) had recurrences, and 5 patients (25%) progressed during the mean follow-up period of 13 months (range, 1-45 months). Five patients (25%) underwent re-resection, 1 patient was lost to follow-up, and 1 patient died secondary to progression of disease. There was 1 intraoperative vascular complication with no sequelae. The cerebrospinal fluid leak rate was 25%, and there were no cases of bacterial meningitis. The incidence of a new permanent neurological complication was 5%. A systematic review of the literature yielded a total of 26 cases of chordomas resected via a completely endoscopic endonasal technique. CONCLUSION: Endoscopic endonasal resection of cranial base chordomas is safe once adequate experience is gained with the technique. This approach provides the potential for, at the least, similar resections compared with traditional cranial base approaches while potentially limiting morbidity. PMID- 19190457 TI - Outcome predictors for intracranial ependymoma radiosurgery. AB - OBJECTIVE: To develop outcome predictors after stereotactic radiosurgery (SRS) in patients with intracranial ependymomas who had received previous fractionated radiation therapy, we compared tumor control, survival, and complications with tumor grade, volume, age of patients, and imaging characteristics. METHODS: We retrospectively reviewed records of 39 consecutive ependymoma patients who underwent SRS for 56 tumors. The median patient age was 22.8 years (range, 2.9 71.1 years). All patients had previous surgical resection of their ependymomas followed by radiotherapy, and 14 patients underwent previous chemotherapy. Twenty five patients had low-grade ependymomas (34 tumors), and 14 patients had anaplastic ependymomas (22 tumors). The median radiosurgery target volume was 3.6 cm (range, 0.1-36.8 cm), and the median margin dose was 15.0 Gy (range, 10-22 Gy). RESULTS: At a median of 23.5 months after SRS (range, 6.1-155.2 months), 25 patients died as a result of metastases (12 patients) or disease progression (13 patients). The overall survival rates after SRS were 60.1, 36.1, and 32.1% at 1, 3, and 5 years, respectively. The progression-free survival rates after SRS at 1, 3, and 5 years were 81.6, 45.8, and 45.8%, respectively, for all grades of ependymomas. Lower histological tumor grade was not significantly associated with better progression-free survival (P = 0.725). Factors associated with an improved progression-free survival included smaller tumor volume and homogeneous tumor contrast enhancement in low-grade ependymomas. CONCLUSION: SRS provides another management option for patients with residual or recurrent ependymomas that have failed surgery and radiation therapy. Predictors of response include smaller volume and homogeneous contrast enhancement. PMID- 19190458 TI - Long-term follow-up of Chiari-related syringomyelia in adults: analysis of 157 surgically treated cases. AB - OBJECTIVE: To determine the long-term outcome of surgically treated Chiari related syringomyelia. METHODS: The medical charts of 157 consecutive surgically treated patients with Chiari-related syringomyelia were retrospectively analyzed. Factors predicting outcome, either clinical or radiological, are discussed, and our results are compared with those of other large series in the literature. RESULTS: The study included 74 men and 83 women (age range, 16-75 years; mean age at surgery, 38.3 years). Pain and sensory disturbance were the most frequent initial symptoms. The average duration of preoperative symptoms was 8.2 years. The follow-up period ranged from 82 to 204 months (median, 88 months). At the end of the study, 99 patients (63.06%) had improved, 48 (30.58%) had stabilized, 9 (5.73%) had worsened, and 1 (0.63%) had died during the postoperative period. Factors predicting improvement or stabilization were young age at the time of surgery and clinical signs of paroxysmal intracranial hypertension. Factors associated with a poor outcome were older age at the time of surgery, arachnoiditis, and a clinical feature of long-tract impairment syndrome. The presence of arachnoiditis or of basilar invagination was associated with poor clinical presentation (P = 0.05 and 0.0001, respectively). The extent of the cyst on postoperative magnetic resonance imaging was a predictor of poor clinical outcome (P = 0.002). CONCLUSION: Our results confirmed that surgery is an effective and safe treatment of Chiari-related syringomyelia, with a 90% chance of long-term stabilization or improvement on average. Surgery should be proposed as soon as possible in patients with clearly progressing clinical features. PMID- 19190459 TI - Fat and neurosurgery: does obesity affect outcome after intracranial surgery? AB - OBJECTIVE: Obesity has been linked to increased morbidity and mortality after some surgical procedures. The purpose of this study was to determine whether obesity affects outcome after general neurosurgery and subarachnoid hemorrhage (SAH). METHODS: Three data sets were analyzed, including a retrospective review of 404 patients undergoing cranial and spinal neurosurgical procedures, a prospective collection of 100 patients with aneurysmal SAH, and data from 3567 patients with aneurysmal SAH who were entered into randomized clinical trials of tirilazad. For each data set, outcome was assessed by mortality, postoperative morbidity, and Glasgow Outcome Scale score. Prognostic factors, including body weight and body mass index, were tested for their effect on these outcomes using multivariable logistic regression. RESULTS: For patients undergoing general cranial and spinal neurosurgery, independent predictors of morbidity and mortality were age, American Society of Anesthesia class, disseminated malignancy, emergency surgery, and increased duration of surgery. For patients with SAH, score on the Glasgow Outcome Scale was associated with age and admission Glasgow Coma Scale score. In the tirilazad data set, multiple factors were associated with score on the Glasgow Outcome Scale, but, as with the other 2 data sets, body weight had no relationship to outcome. CONCLUSION: Obesity may have less effect on the outcome of patients with mainly cranial neurosurgical disease and aneurysmal SAH than it does on patients undergoing other types of surgery. PMID- 19190460 TI - Epilepsy surgery in children with electrical status epilepticus in sleep. AB - OBJECTIVE: Pediatric epilepsy surgery candidates with unilateral congenital or early-acquired brain lesions may present with refractory seizures and generalized electroencephalographic features such as electrical status epilepticus in sleep (ESES). The purpose of our study was to review the clinical presentation, neuroimaging findings, and outcome in a series of children with unilateral brain lesions and ESES undergoing resective surgery for refractory epilepsy. METHODS: A total of 415 consecutive patients younger than 18 years of age undergoing video electroencephalographic evaluation and epilepsy surgery at Cleveland Clinic were reviewed for ESES, an underlying pathological lesion, and outcome after surgery. RESULTS: Eight patients were included. All patients presented with medically refractory epilepsy, hemiparesis, and developmental delay. The pathogenesis was perinatal infarction in 7 patients and malformation of cortical development in 1 patient. Preoperative electroencephalography demonstrated generalized interictal spikes, electroencephalographic seizures, and ESES in all cases. Age at the time of surgery ranged from 3 to 14 years. Six patients underwent hemispherectomy, and 2 patients underwent focal resection. Six patients became seizure-free after resection. Two patients with functional hemispherectomy continued to have rare seizures, but were much improved. These patients also had perinatal infarctions in the hemisphere contralateral to the resection, possibly indicating a less beneficial outcome. Postoperative electroencephalography demonstrated resolution of generalized interictal discharges and ESES in all. Formal pre- and postoperative neuropsychological testing showed overall improvement of age equivalent scores. CONCLUSION: Children with unilateral brain lesions and seizures may become seizure-free after epilepsy surgery, even if the preoperative electroencephalogram shows generalized ESES. The lesion occurring early in life and the location of the lesion may play a role in the development of ESES. Cognitive impairment may be aggravated by the persistence of ESES. Preliminary developmental data in this small sample suggest that termination of seizures and possibly of ESES by epilepsy surgery may have developmental benefits. PMID- 19190461 TI - Recurrent disc herniation and long-term back pain after primary lumbar discectomy: review of outcomes reported for limited versus aggressive disc removal. AB - OBJECTIVE: It remains unknown whether aggressive disc removal with curettage or limited removal of disc fragment alone with little disc invasion provides a better outcome for the treatment of lumbar disc herniation with radiculopathy. We reviewed the literature to determine whether outcomes reported after limited discectomy (LD) differed from those reported after aggressive discectomy (AD) with regard to long-term back pain or recurrent disc herniation. METHODS: A systematic MEDLINE search was performed to identify all studies published between 1980 and 2007 reporting outcomes after AD or LD for a herniated lumbar disc with radiculopathy. The incidence of short- and long-term recurrent back or leg pain and recurrent disc herniation was assessed from each reported LD or AD cohort and the cumulative incidence compared. RESULTS: Fifty-four studies (60 discectomy cohorts) met the inclusion criteria, reporting the outcomes of 13 359 patients after lumbar discectomy (LD, 6135 patients; AD, 7224 patients). The reported incidence of short-term recurrent back or leg pain was similar after LD (mean, 14.5%; range, 7-16%) and AD (mean, 14.1%; range, 6-43%) (P < 0.01). However, more than 2 years after surgery, the reported incidence of recurrent back or leg pain was 2.5-fold less after LD (mean, 11.6%; range, 7-16%) compared with AD (mean, 27.8%; range, 19-37%) (P < 0.0001). The reported incidence of recurrent disc herniation after LD (mean, 7%; range, 2-18%) was greater than that reported after AD (mean, 3.5%; range, 0-9.5%) (P < 0.0001). CONCLUSION: Review of the literature demonstrates a greater reported incidence of long-term recurrent back and leg pain after AD but a greater reported incidence of recurrent disc herniation after LD. Prospective, randomized trials are needed to firmly assess this possible difference. PMID- 19190462 TI - Radiosurgery with the world's first fully robotized Leksell Gamma Knife PerfeXion in clinical use: a 200-patient prospective, randomized, controlled comparison with the Gamma Knife 4C. AB - OBJECTIVE: The world's first Leksell Gamma Knife PerfeXion (Elekta Instrument AB, Stockholm, Sweden) for radiosurgery of the head and neck became operational at Timone University Hospital in Marseille on July 10, 2006. To allow strict evaluation of the capabilities, advantages, disadvantages, and limitations of this new technology, patients were enrolled in a prospective, randomized trial. METHODS: In 66 working days, between July 10 and December 20, 2006, 363 patients were treated by gamma knife surgery at Timone University Hospital, Marseille. Of these patients, 200 were eligible for the comparative prospective study (inclusion criteria were informed consent obtained, tumor or vascular indication, and no previous radiosurgery or radiotherapy). In accordance with the blinded randomization process, 100 patients were treated with the Leksell Gamma Knife 4C (Elekta Instrument AB) and Gamma Knife 100 (Elekta Instrument AB) with the Leksell Gamma Knife PerfeXion. Dose planning parameters, dosimetry measurements on the patient's body, workflow, patient comfort, quality assurance procedure, and a series of other treatment-related parameters were systematically and prospectively evaluated in both arms of the trial. RESULTS: No technical failure of the treatment procedure was encountered. The new dose-planning system led to the use of composite shots in 39.4% of the patients. The median number of different collimator sizes used was larger with the PerfeXion than with the 4C (2 and 1, respectively). The mean number of isocenters used was lower (10.67 and 13.08, respectively). The median total treatment time was significantly shorter with the PerfeXion (40 and 60 minutes, respectively), but there was no significant difference in the median radiation time (34.02 and 33.40 minutes, respectively). The procedure was performed using only a single run in 98.99% of the PerfeXion cases and in 42% of the 4C cases. Collision risk on the 4C forced us to change the frame gamma angle for at least 1 shot in 24% of the patients and led to treatment in manual mode for at least 1 shot in 21% of the patients. Collision risk requiring technical adaptation did not occur with the PerfeXion. In 1 patient treated with the PerfeXion, the system required a direct collision check. In terms of dose to structures outside the target area, the PerfeXion delivers 8.2 times less to the vertex, 10 times less to the thyroid, 12.9 times less to the sternum, and 15 times less to the gonads. CONCLUSION: Our prospective study indicates that procedures with the PerfeXion were collision-free, even with very eccentric lesions (e.g., multiple metastases). The duration of the surgical procedure, the amount of time required for nurse, physicist, and physician intervention on the machine, and the duration of the quality assurance procedure were all shown to be dramatically reduced with the PerfeXion gamma knife. Patient protection is greatly improved with the PerfeXion. In our experience, the technological advances of the Leksell Gamma Knife PerfeXion will make a very significant contribution to future progress in head and neck radiosurgery. PMID- 19190463 TI - Pitavastatin suppresses formation and progression of cerebral aneurysms through inhibition of the nuclear factor kappaB pathway. AB - OBJECTIVE: Recent investigations strongly suggest that the pathophysiology of cerebral aneurysms (CA) is closely associated with chronic inflammation in vascular walls. Nuclear factor kappaB (NF-kappaB) has a key role in the formation and progression of CAs. Because statins exert anti-inflammatory effects in various vascular diseases, we investigated the effect of pitavastatin on NF kappaB activation and CA formation in experimentally induced CAs in rats. METHODS: CAs were induced in Sprague-Dawley rats with or without administration of pitavastatin (4 mg/kg/d orally). Size, change of internal elastic lamina, and media thickness of induced CAs were measured in both groups after aneurysm induction. The effects of pitavastatin on NF-kappaB activation in aneurysmal walls were examined by immunohistochemistry and gel shift assay. Expression of downstream genes was analyzed by quantitative polymerase chain reaction and immunohistochemistry. To examine whether pitavastatin has a suppressive effect on preexisting CAs, pitavastatin administration started 1 month after aneurysm induction. RESULTS: Pitavastatin treatment significantly prevented CA progression (P < 0.01) and NF-kappaB activation in aneurysmal walls. Expression of monocyte chemotactic protein-1, vascular cell adhesion molecule-1, interleukin-1beta, inducible nitric oxide synthase, and matrix metalloproteinase-9 in aneurysmal walls was also inhibited by pitavastatin. Pitavastatin treatment led to media thickening in preexisting CAs. CONCLUSION: Pitavastatin has a suppressive effect on CA progression through the inhibition of NF-kappaB activation in aneurysmal walls. Moreover, pitavastatin treatment can cause the regression of degenerative changes in preexisting CA walls. Pitavastatin is a promising candidate for a novel preventive agent against subarachnoid hemorrhage. PMID- 19190464 TI - Treatment of experimental aneurysms with an embolic-containing device and liquid embolic agent: feasibility and angiographic and histological results. AB - OBJECTIVE: To assess the feasibility and analyze angiographic and histological results of experimental aneurysms treated with an embolic-containing device (ECD) and to test a liquid embolic agent (LEA), N-butyl cyanoacrylate. METHODS: Four experimental large bifurcation aneurysms and 1 large sidewall aneurysm were created in 5 dogs. These venous pouch aneurysms were occluded endosaccularly with an ECD and N-butyl cyanoacrylate. Angiographic and/or histopathological data were analyzed at 1 day (bifurcation), 1 week (bifurcation), 4 months (sidewall), 6 months (bifurcation), and 12 months (bifurcation). RESULTS: Aneurysm dimensions were 16 to 18 mm in length, 6 to 8 mm in width, and a neck measuring 6 to 8 mm. Immediately after the procedure, 2 of 4 bifurcation aneurysms were 100% occluded. In one case, glue had spilled into the parent artery. One occluded aneurysm recanalized at 6 months. One occluded aneurysm remained closed at 12 months. Histopathological analysis of the 6- and 12-month specimens showed adherence of glue to the aneurysm wall, no appreciable inflammatory response to the ECD, and aneurysm wall fibrosis with adventitial chronic inflammation. The aneurysm necks were covered by neointima with the presence of endothelialization. CONCLUSION: Within the limitations of this experimental study, treatment of large, wide necked aneurysms with the ECD and LEA may be feasible. Suboptimal technique and ECD geometry can cause leakage of LEA into the parent vessel or incomplete apposition of the ECD/glue to the aneurysm wall. However, the ECD and glue injection technique did achieve complete occlusion in 1 aneurysm that persisted 1 year later. The histopathological findings in this instance are moderately encouraging. Further investigations of an ECD with N-butyl cyanoacrylate or another LEA are warranted. PMID- 19190465 TI - Averroes in The school of Athens: a Renaissance man and his contribution to Western thought and neuroscience. AB - The European Renaissance was a revolutionary movement in human history. In this era of the rebirth of humanity, a great number of valuable artworks were created. One of these masterpieces is the magnificent wall painting The School of Athens, by Raphael, which is also known as a "visualization of knowledge." Raphael depicted almost all ancient Greek philosophers in this fresco. The painting displays one noticeable figure among the scholars: the philosopher/scientist Averroes from Andalusia. Western thought was greatly influenced by Averroes' philosophy. Averroes was as extraordinary a physician as he was a philosopher. His medical works presented novel concepts such as the discovery of the photoreceptor function of the retina, describing symptoms and signs of Parkinson's disease, and proposing a cardiac/vascular origin of cerebral stroke. In this article, we aimed to introduce Averroes' works and his contribution to neuroscience in the context of art, framed particularly with Raphael's masterwork The School of Athens. PMID- 19190467 TI - Taking care of those who need us most. PMID- 19190468 TI - Planning ahead for better outcomes: preparation for joint replacement surgery begins at home! AB - For 15 years, a small community hospital has sent registered nurses into the home to do a preoperative visit with patients planning joint replacement surgery. These visits help the patient by providing a better understanding of what to expect from the preadmission visit through the recovery period. It also ensures a safe environment for the recovery process whether at home or alternative environment with family/friends or at a short-term rehabilitation facility. Its success in patient and nurse satisfaction has not only saved it from cost cutting measures but also allowed it to expand over the years to include follow-up home visits and now includes patients undergoing shoulder replacement. This was a great undertaking for a hospital of this size in 1993 and is one of the few, if not the only, such home visit programs in Massachusetts, doing both pre and postoperative visits, funded by a hospital, utilizing nursing hours. PMID- 19190470 TI - Recurrent popliteal cyst in an adult: a case report and review. AB - Popliteal cysts are typically secondary to an intra-articular pathology in adults but may occur as primary lesions unrelated to knee injury. Careful examination of any mass forming on the posterior aspect of the knee is needed to differentiate benign from malignant conditions. Asymptomatic cysts are often treated conservatively, whereas surgical intervention is recommended for troublesome cysts and for those related to intra-articular derangements. Surgical intervention focused on amelioration of intra-articular derangements and the communication between the knee and the popliteal cyst results in cyst recurrence of less than 5%. A report of a recurrent popliteal cyst, in spite of aggressive surgical intervention, is presented. PMID- 19190472 TI - Wildfire disaster leads to facilities evacuation. AB - Disaster preparedness in the healthcare setting traditionally focuses on accepting the capacity surge and influx of patients in time of crisis. The concept of a hospital evacuation is given little thought, and minimal time is spent practicing even though healthcare organizations have identified a hazard vulnerability that may require a full-scale evacuation. One healthcare organization recently pushed its staff to stretch its idea of disaster preparedness when it evacuated a community hospital and skilled nursing facility to ensure the safety of its patients and staff. The success of the evacuation was attributed to communication, preplanning, and leadership within the organization. PMID- 19190474 TI - Podcasts, blogs, and webinars. PMID- 19190475 TI - Understanding the health literacy of America: results of the National Assessment of Adult Literacy. AB - Health literacy refers to an individual's ability to understand healthcare information to make appropriate decisions. Healthcare professionals are obligated to make sure that patients understand information to maximize the benefits of healthcare. The National Assessment of Adult Literacy (NAAL) provides information on the literacy/health literacy levels of the U.S. adult population. The NAAL is the only large-scale survey of health literacy. The results of the NAAL provide information on literacy/health literacy and the relationship between background variables and literacy/health literacy. Multiple variables with potential for a relationship with literacy/health literacy were chosen for the NAAL including, but not limited to, education, language, race, gender, income, overall health, seeking health information, and health insurance. PMID- 19190477 TI - Treating clients with a deadly poison. AB - Botulinum toxin (Botox) injections are a highly sought-after elective treatment for temporary relief of some facial lines. Botox is also a highly useful therapeutic agent, approved for use in treating a variety of health conditions. However, Botulinum toxin is a poison responsible for botulism, and, regardless of the purpose for Botox injections, there are serious considerations about safety and adverse effects related to its use that are important for nurses to know. In this article, the relationship between botulism and therapeutic use of Botox is identified. An overview of information about Botox includes particulars related to safe use and Federal Drug Administration warnings and suggestions for patient education. PMID- 19190479 TI - Collateral ligament interarticular avulsion fracture. PMID- 19190480 TI - Lumbar flexion and dynamic EMG among persons with single level disk herniation pre- and postsurgery with radicular low-back pain. AB - OBJECTIVE: The paraspinal muscles often fail to relax on forward flexion in many persons with low-back pain. The goal of this prospective study was to determine whether this abnormal lack of a flexion-relaxation phenomenon corrects after lumbar diskectomy for symptoms of radiculopathy with low-back pain. DESIGN: Electromyographic testing was performed on 17 patients before and 30 days after lumbar diskectomy. RESULTS: Although pain improved significantly (P < 0.05), the flexion-relaxation phenomenon did not improve. CONCLUSIONS: Failure to recover muscle relaxation while pain is relieved suggests another mechanism for paraspinal activity. PMID- 19190481 TI - Pressure ulcers in spinal cord injury: an unusual site and etiology. AB - Case series of pressure ulcers in patients with spinal cord injury after inappropriate and prolonged application of graduated compression stockings are presented. Three adult patients with spinal cord injury were fitted with graduated compression stockings in the acute phase to prevent thromboembolism. Acute hospital staff did not monitor the skin for potential pressure ulcer because of the graduated compression stockings. Grade II pressure ulcers developed on the toes in all three cases. All pressure ulcers were managed conservatively and healed without complication. The manufacturers of graduated compression stockings did not include information about the need to monitor patients with sensory impairment for pressure ulcers. It is suggested that the manufacturers of graduated compression stockings include information about the need to monitor patients with sensory impairment for skin breakdown. When graduated compression stockings are applied to patients with sensory impairment, including spinal cord injury, staff needs to be vigilant for pressure ulcers occurring at the extremes of the graduated compression stockings, in addition to the more common areas where pressure ulcers occur. PMID- 19190482 TI - Gait changes after tendon functional surgery for equinovarus foot in patients with stroke: assessment of temporo-spatial, kinetic, and kinematic parameters in 177 patients. AB - OBJECTIVE: In patients with hemiplegic stroke, equinovarus foot is one of the most frequent deformities. Outcome evidence for surgical correction of equinovarus foot is scarce, and results are usually assessed only clinically. Moreover, concerns about possible loss of function after elongation of the plantar flexor muscles are still at issue. The objective of this study was to verify if surgical correction of equinovarus foot can improve gait speed and function. DESIGN: We used a retrospective, nonrandomized design. One hundred seventy-seven chronic hemiplegic patients who underwent surgical correction of equinovarus foot were evaluated before and 1 yr after surgery. Outcome measures were walking handicap score, temporal-spatial parameters, gait kinematics and kinetics, and paretic propulsion. RESULTS: After surgery, walking handicap and temporal-spatial parameters significantly improved, as did ankle kinematic data and gait kinetic data. Patients' gait at follow-up was faster, with a more normal base of support and with better foot advancement. Paretic propulsion increased significantly after surgery, even if ankle power at push-off was reduced. We also observed a low complication rate. CONCLUSIONS: Surgical correction of equinovarus foot deformity in patients with stroke is a safe and effective procedure. Even if the power generation at the ankle decreased, overall gait function and parameters improved after surgery. PMID- 19190483 TI - Voluntary activation of the knee extensors in chronic poststroke subjects. AB - OBJECTIVE: To assess the extent to which knee extensor muscle weakness in subjects with chronic mild to moderate poststroke hemiparesis is caused by a decreased voluntary activation. DESIGN: Forty community dwelling and ambulant men and women (mean age, 59.8 +/- 5.5 yrs) with residual hemiparesis (19.2 +/- 8.5 mos poststroke) were tested. Torque measurements were performed on a computerized dynamometer and the superimposed electrical stimulation technique was used to assess voluntary activation of the knee extensors in both the paretic and the nonparetic lower limbs. RESULTS: The mean voluntary activation ratio of the knee extensors in the nonparetic and paretic leg was 0.97 +/- 0.04 and 0.86 +/- 0.13, respectively. Subjects who had a greater relative weakness, implying a more pronounced poststroke impairment, also had lower voluntary activation ratios. The mean percentage difference in total torque between the nonparetic and the paretic knee extensors after the electrical stimulation was 36.4% +/- 17.0%. CONCLUSIONS: Paretic knee extensor muscle weakness in chronic poststroke subjects is only partially explained by a reduced voluntary activation ability, indicating that other neuromuscular structural or functional factors contribute to poststroke hemiparetic muscle weakness. PMID- 19190484 TI - Incidental findings during functional magnetic resonance imaging: ethical and procedural issues. AB - Incidental findings during fMRI: ethical and procedural issues.This purpose of this report was to describe the discovery of an incidental finding during functional magnetic resonance imaging in a child of typical development. During the completion of a functional magnetic resonance imaging protocol, an abnormality was noted by a board-certified neuroradiologist that was identified as a benign developmental arachnoid pouch. Significant ethical concerns were apparent with the discovery of an incidental finding, which included how to address the likelihood of incidental findings in the consent and assent process, how to disclose the incidental findings to the subject's parents and primary care provider, and how to minimize subject and parental anxiety. PMID- 19190485 TI - Measurement properties of fine motor scale of Peabody developmental motor scales second edition: a Rasch analysis. AB - OBJECTIVE: To investigate the measurement properties (including rating scale performance, unidimensionality, and differential item functioning) of the fine motor scale of the Peabody Developmental Motor Scales-Second Edition in children, by using the Rasch analysis. DESIGN: A total of 419 children (including 342 typically developing children and 77 children with fine motor delays or difficulties) were recruited in Taiwan for this prospective study. Each child was evaluated with the Peabody Developmental Motor Scales-Second Edition that consists of 26-item grasping and 72-item visual-motor integration subtests. Partial credit Rasch analysis was used for all analyses. RESULTS: The Rasch analysis indicated that middle rating category for 19 grasping and 52 visual motor integration items could be collapsed to allow only dichotomous response categories. Item fit analysis and principal component analysis suggested that the unidimensionality of the grasping and visual-motor integration subtests could be achieved after removal of two grasping and eight visual-motor integration misfitting items. All but 13 items in the composite scale could form a unidimensional construct of overall fine motor ability. Furthermore, only a few items were found to show differential item functioning across sex (ten items) or fine motor status (seven items). However, significant ceiling effects were found in the Peabody Developmental Motor Scales-Second Edition subtests and composite scale when applied to these typically developing children. CONCLUSIONS: Our results suggest grounds for the revision of the Peabody Developmental Motor Scales-Second Edition in a subsequent edition. Simplifying the rating scales and reducing the misfitting items in the subtests and composite scales might result in a unidimensional assessment of children's fine motor ability. Clinicians and researchers could use the reduced Peabody Developmental Motor Scales-Second Edition as a criterion-referenced outcome measure to document changes; however, further work is needed to reduce the ceiling effects. PMID- 19190487 TI - Hydatid cyst disease of the spine. PMID- 19190486 TI - Use of physical and occupational therapy by Medicare beneficiaries within five conditions: 1994-2001. AB - OBJECTIVE: To examine the use of physical therapy and occupational therapy among Medicare beneficiaries nationwide before and after the 1997 Balanced Budget Act, which introduced prospective payment for rehabilitation services. DESIGN: We analyzed responses from the longitudinal Medicare Current Beneficiary Survey, merged with Medicare claims, to track physical therapy and occupational therapy rates and intensity (units of service) from 1994 through 2001. This observational study focused on elderly and disabled Medicare beneficiaries within five conditions: stroke, acute myocardial infarction, chronic obstructive pulmonary disease, arthritis, and lower-limb mobility problems. We used cubic smoothing spline functions to describe trends in service intensity over time and generalized estimating equations to assess changes in service intensity. RESULTS: Controlling for demographic characteristics, adjusted mean level of physical therapy and occupational therapy intensity rose significantly between 1994 and 2001 for all five conditions. Service intensity leveled off in 1999 for occupational therapy and 2000 for physical therapy. With few exceptions, physical therapy and occupational therapy intensity was not significantly associated with patients' demographic characteristics. CONCLUSIONS: Medicare beneficiaries with conditions that can potentially benefit from physical therapy or occupational therapy or both continued to get these services at similar-and sometimes increasing-intensity during years after passage of the Balanced Budget Act. PMID- 19190488 TI - Quality of life of men and women with osteoarthritis of the hip and arthroplasty: assessment by WOMAC questionnaire. AB - OBJECTIVE: The aim of this study was to investigate the differences in quality of life between men and women in preoperative and postoperative period after hip arthroplasty because of severe hip osteoarthritis. DESIGN: This is a prospective study of 160 patients (average age, 61.7 yrs), 92 women and 68 men, with a diagnosis of osteoarthritis of the hip who underwent total hip arthroplasty. All patients completed the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) questionnaires that measured health-related quality of life preoperatively, at discharge, and 6-wk postoperatively. To establish the occurrence of differences between men and women in preoperative and postoperative period after arthroplasty, Student's t test and multivariate logistic regression analysis were used. RESULTS: The WOMAC global score was a significant predictor (the better the rather men) preoperatively (P < 0.01) and 6 wks after total hip arthroplasty (P < 0.001). Global WOMAC score was significantly better in men than in women preoperatively (t = 4.02; P < 0.001) and 6 wks after arthroplasty (t = 3.42; P < 0.001). CONCLUSIONS: These results suggest that men with severe osteoarthritis of the hip have better quality of life than do women preoperatively and 6 wks after hip arthroplasty. These findings would be important for improving quality of care of our patients. PMID- 19190489 TI - Noninvasive determination of the tension-time index in Duchenne muscular dystrophy. AB - OBJECTIVE: Noninvasive determination of the tension-time index of the inspiratory muscles (TT MUS) can identify predisposition to respiratory muscle fatigue in neuromuscular disease. We correlated TT MUS with age and extent of need of ventilator use for patients with Duchenne muscular dystrophy. DESIGN: Maximal inspiratory pressure, inspiratory pressure 0.1 sec after the onset of inspiration (P 0.1), and the breathing pattern during spontaneous breathing were measured in 46 subjects with Duchenne muscular dystrophy and in 46 healthy males of the same age. TT MUS (TT MUS = T I/T TOT x P I/MIP) was determined by calculating P I from P 0.1 (P I = 5 x P 0.1 x T I). The data were compared with normal values and related to age and ventilator use. RESULTS: TT MUS was significantly higher in the entire Duchenne muscular dystrophy group than in controls (0.21 +/- 0.11 vs. 0.06 +/- 0.02, P < 0.001) and increased with age in the patients (P < 0.001). TT MUS was significantly higher in subjects ventilated 8-20 hrs per day than in nocturnal-only users and in patients ventilated >20 hrs per day than in those ventilated 8-20 hrs per day (P < 0.001). TT MUS surpassed 0.23 in 95% of subjects ventilated 8-20 hrs. TT MUS exceeded 0.37 in 95% of individuals ventilated >20 hrs per day, whereas it was less than this value in 95% of patients ventilated 8 20 hrs. CONCLUSIONS: TT MUS increases significantly with disease progression and is a sensitive indicator of risk of inspiratory muscle fatigue and ventilator use. Longitudinal determination of TT MUS in patients with Duchenne muscular dystrophy may help to justify the extent of need for mechanical ventilation. PMID- 19190490 TI - Comparison of two automated sphygmomanometers for use in the office setting. AB - OBJECTIVE: To compare readings obtained using two automated blood pressure (BP) recording devices, BpTRU and Omron 907, in a clinical setting. METHODS: Two series of 50 patients attending a hypertension unit for 24-h ambulatory BP monitoring had BP recorded either every 1 or every 2 min using BpTRU and Omron 907 devices with the order of the measurements being randomized. RESULTS: No significant differences for systolic BP between the mean readings taken using the BpTRU or Omron 907 recorders at either 1 or 2 min were observed. Diastolic BP readings were similar using the 1-min interval setting but were 5.0 mmHg lower for the Omron 907 (P<0.001) when readings were taken at 2-min intervals. CONCLUSION: BP measurements can be made in the clinical setting using either the BpTRU or Omron 907 automated sphygmomanometers. PMID- 19190491 TI - Conventional and 24-h ambulatory blood pressure as independent predictors of elastic arterial properties. AB - OBJECTIVE: No population study investigated whether 24-h ambulatory blood pressure (ABP) predicts distensibility of the elastic common carotid (DCar) and the muscular femoral (DFem) arteries over and beyond conventionally measured blood pressure (CBP). METHODS: At baseline, we measured CBP and 24-h ABP in 1063 randomly recruited participants (mean age, 44.3 years). CBP was the average of five consecutive readings obtained by trained observers at the participants' homes. We measured arterial distensibility by a wall-tracking ultrasound system, 21 months after CBP and ABP (5-95th percentile interval range, 13-33 months). RESULTS: Compared with men, women (49.2%) had higher (P<0.03) DCar (24.7 vs. 23.3 x 10(-3)/kPa) and higher DFem (10.6 vs. 9.2 x 10(-3)/kPa). In multivariate adjusted models, including both CBP and ABP and stratified by sex, DCar was negatively related to systolic, diastolic, and mean arterial CBP in both sexes, and to diastolic ABP in women. DFem was inversely correlated with diastolic ABP in both sexes and with systolic and mean arterial ABP in men. Moreover, DFem was also negatively correlated with systolic and mean arterial CBP in men. In most instances, pulse pressure on CBP or ABP measurement did not predict DCar or DFem. No evidence of influential collinearity between CBP and ABP was observed. CONCLUSION: Depending on vascular territory, there is competition between highly standardized CBP and ABP in predicting DCar and DFem. These findings show that CBP under standardized conditions, and subject to rigorous quality control, is equally predictive of the elastic properties of large arteries as ABP. PMID- 19190492 TI - Home sphygmomanometers: validation versus accuracy. AB - OBJECTIVE: The validation and accuracy of sphygmomanometers are important issues in the home sphygmomanometer market and in clinical practice. The aims of this study are to assess the frequency of validated home sphygmomanometers and to evaluate the relationship between the validation and accuracy of the devices. METHODS: Eight hundred and ninety-six home sphygmomanometers were brought by individuals to the University Hospital Hypertension Clinic for this study, and 870 (97%) of the devices were in adequate working condition and suitable for analysis. The relationship between accuracy and validation was investigated only in automated sphygmomanometers (n = 554). RESULTS: Both the manufacturer and model were known in 500 devices (90%), and these devices were eligible for evaluation of validation. We found 74 devices listed in the dabl Educational Trust website. Twenty-two of the 74 (30%) devices were validated. Four hundred (72%) of the 554 automated sphygmomanometers were inaccurate. The frequency of accuracy was higher among validated devices compared with nonvalidated devices (68 vs. 15%) (P<0.01). CONCLUSION: Our study showed that the frequency of accurate devices was higher among validated sphygmomanometers compared with nonvalidated sphygmomanometers. To our knowledge, our study is the first to demonstrate the relationship between accuracy and validation of home sphygmomanometers. The frequency of device-related errors can be decreased by training patients and supervising the blood pressure device market. Nonprofit organizations can help patients to overcome some of the problems in the blood pressure device market. PMID- 19190493 TI - Adapting standards of care under extreme conditions. AB - The filing of criminal charges against a group of clinicians in New Orleans for failure to meet expected standards of care following the hurricanes of 2005 made the growing concern among health professionals about care provided during extreme emergencies or disasters all too real. Questions about what may lead to censure, penalties from licensing boards, or lawsuits have come from nurses, physicians, and many other licensed health professionals. A panel convened by the American Nurses Association that included representatives of medical, public health, hospital, and government agencies considered the ethical, professional, and practical aspects of meeting standards of care in such circumstances. Clinicians are reminded that in emergencies, it is only the circumstances that change (perhaps radically); neither the individual's professional competency nor the basic professional standard of care is different. In making prioritized decisions under such circumstances, the individual's ethical framework is utilitarian, and there are 3 areas for action, even when some routine tasks are set aside: maintain worker and patient safety; maintain airway, breathing, and circulation; and establish or maintain infection control. Policy recommendations such as state legislation for the adoption of comprehensive immunity for volunteer health care workers, and the establishment of a medical review panel as arbitration board are also suggested. The resulting white paper summarizes the issues and provides guidance to individual professionals, institutions in which they work, and emergency planners. PMID- 19190494 TI - Educational inequalities in cancer incidence in Turin, Italy. AB - The objective of this study was to investigate the relationship between cancer incidence and socioeconomic status, and to examine the temporal trends in social inequalities in cancer risk. Educational differentials in the incidence of cancer (25 sites) among adult residents of Turin (Italy) were examined using data from the Turin Longitudinal Study and the Piedmont Cancer Registry. The relationship between cancer incidence and educational level was evaluated over three 5-year periods between 1985 and 1999 using Poisson models. An estimated 17% of malignancies among men in the low-educational group were attributable to education, whereas women with a low educational level were slightly protected. Less-educated men had higher risks of upper aero-digestive tract, stomach, lung, liver, rectal, bladder, central nervous system and ill-defined cancers, and lower risks of melanoma, kidney and prostate cancers. Women with lower educational levels were at higher risk of stomach, liver and cervical cancers, whereas they were less likely to be diagnosed with melanoma, ovarian and breast cancers. For most sites, the educational gradient in risk did not vary substantially over time. The educational inequalities in cancer incidence observed in this cohort appear similar in magnitude and direction to socioeconomic inequalities found in other Western countries; for some cancer sites results partly differ from the results of other studies, and require further investigation. A thorough understanding of the relative burden of well-documented causes of social inequalities in cancer risk is essential to address preventive measures and to direct future research on unexplained social differences. PMID- 19190495 TI - Seizure or syncope: the diagnostic value of serum creatine kinase and myoglobin levels. AB - OBJECTIVE: To determine if the serum levels of creatine kinase and myoglobin can be used to differentiate the grandmal tonic-clonic seizure and syncope activities in the emergency department (ED). METHODS: Consecutive patients over 16 years old who presented to an ED of a tertiary care hospital with a witnessed tonic-clonic seizure activity and a history of a transient loss of consciousness with normal neurological exams in the ED were selected to either seizure or syncope groups. Patients with an unclear history of seizure or syncope with more than 4 h of the activity and with any conditions that could elevate creatine kinase and myoglobin levels were excluded. Serum samples were drawn at presentation and at the fourth hour of the event. RESULTS: Thirty-seven syncope and 26 generalized tonic-clonic seizure patients with a definite history were assigned to study groups. There was not a statistically significant difference in the time of drawing of the first serum sample among groups. No statistically significant differences were determined with the first samples of creatine kinase and myoglobin for both groups. Serum levels of creatine kinase drawn at the fourth hour of the activity were significantly higher in favor of the seizure group. However, myoglobin levels were insignificant at the fourth hour. CONCLUSION: Serum creatine kinase measured at the fourth hour of loss of consciousness may be a potentially useful laboratory test to differentiate tonic-clonic seizure from syncope. Patently, it requires and warrants further study. PMID- 19190496 TI - Lamivudine monotherapy for chronic hepatitis B infection with acute exacerbation revisited. AB - BACKGROUND: Acute exacerbation (AE) of chronic hepatitis B virus (HBV) infection in cancer chemotherapy patients and in organ transplant recipients receiving immunosuppressants may cause catastrophe and high mortality. Hence, immediate treatment with nucleoside analogues for such patients has become a consensus. Anti-HBV therapeutic trials in Asia have shown that AE of chronic hepatitis B (CH B) may result in increased sustained remission (SR) rate with lamivudine monotherapy. Nonetheless, AE episodes in CH-B patients may evolve uneventfully and lead to spontaneous remission. Thus, the policy of immediate anti-HBV therapy for AE patients reaches an impasse. Once treatment is initiated, life long HBV suppression may be necessary. OBJECTIVE: To determine whether lamivudine monotherapy during an AE of CH-B results in an increase in SR compared with no therapy. METHODS: A cohort of 154 CH-B patients seropositive for hepatitis B e antigen with AE formed the study group. This included 102 cases receiving a nationwide therapeutic trial of 18-month lamivudine monotherapy that were compared with 52 cases with no therapy. All were observed for at least 30 months, which encompassed the 18-month on treatment period and a 12-month posttreatment follow-up. RESULTS: No significant increase was observed in the SR rate in the lamivudine treatment group compared with the spontaneous remission rate in the untreated patients (P=0.782, Fisher's exact test). CONCLUSION: AE does not increase the SR rate during 18-month lamivudine monotherapy. Immediate lamivudine therapy for AE patients is not justified as mandatory. The policy should be only applied to AE patients with impending liver failure. PMID- 19190497 TI - Massive bleeding from ectopic varices in the postpartum period: rare but serious complication in women with portal hypertension. AB - Portal hypertension (PHT) often leads to collateralization of blood flow through variceal vessels that shunt blood from the portal to the systemic circulation. Life-threatening bleeding from esophageal and ectopic varices often complicates severe PHT. Increase in PHT occurs during the last stages of the second trimester of pregnancy and is associated with increased risk of PHT bleeding in the later stages of pregnancy. In this report, we present two rare cases of pregnant women with PHT, who had postpartum bleeding from very uncommon sites. The first had a rupture of an intra-abdominal varix and the second had two episodes of bleeding from abdominal wall varices, after two emergent cesarean sections, in two consecutive pregnancies. On the basis of a literature review, we constructed an algorithm that includes instructions on how to handle women with PHT during the various stages of pregnancy and labor. PMID- 19190498 TI - Unintentional video capsule bronchoscopy. PMID- 19190499 TI - Improvable quality of diagnostic assessment of colorectal cancer in southern Netherlands. AB - OBJECTIVE: To determine the extent of guideline implementation of the diagnostic approach in patients with colorectal cancer (CRC) in southern Netherlands in 2005, with special focus on colonoscopy. METHODS: Data were extracted from the medical records for a random sample of 257 colon and 251 rectal cancer patients newly diagnosed in 2005 and recorded from the Eindhoven Cancer Registry. Adherence to guidelines was determined for diagnostic assessment. Multivariable logistic regression analysis was conducted to assess determinants of complete colonoscopy. RESULTS: Diagnostic assessment was carried out mainly by internists (50%) and gastroenterologists (36%). Colonoscopy was performed in 83% of patients with proximal/transverse colon cancer, 55% of those with distal colon cancer, and 65% of those with rectal cancer. A tumour biopsy was taken for 84% of colon and 93% of rectal tumours. Colonoscopy completeness was lower for patients with comorbidity, obstructing tumours, and patients with poor bowel preparation. Abdominal ultrasound was performed for 72% of colon and 52% of rectal cancer patients and a thoracic radiography of over 80% of CRC patients. Computed tomography (CT) of the abdomen was performed in over half of the colon cancer patients and a pelvic CT scan or magnetic resonance imaging in 36% of rectal cancer patients. CONCLUSION: Improvements in adherence to diagnostic guidelines for CRC appear possible, especially in the performance of imaging procedures. Among patients where complete visualization of the colon was not feasible with colonoscopy, imaging techniques such as virtual CT might be of added value in the near future. PMID- 19190500 TI - Liver steatosis in patients with chronic hepatitis B infection: host and viral risk factors. AB - BACKGROUND: Liver steatosis (LS) and chronic infection with hepatitis B virus (HBV) are two common causes of chronic liver disease in Iran. LS is also a common histopathological feature of chronic hepatitis B (CHB). Association of chronic HBV infection and LS has not been extensively studied. AIM: We investigated the prevalence of histological evidence of LS in a group of patients with chronic HBV infection undergoing liver biopsy and compared clinical data, laboratory features, and severity of hepatic fibrosis between patients both with and without steatosis. METHODS: A total of 132 patients with CHB, undergoing liver biopsy for diagnostic purposes over a 2-year period were enrolled in this study. Clinical, biochemical, and histological factors that might have any kind of association with the presence of steatosis were evaluated. RESULTS: Of the 132 patients with a liver biopsy, steatosis was present in 56 (42.4%) of the patients, of whom 36 (64%) had grade 1, 14 (25%) grade 2, and six (10.7%) grade 3. Our data showed that LS is not associated with age, sex, HBeAg, viral load, amount of fibrosis, serum cholesterol level, aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase. Although body mass index, serum triglyceride, fasting blood glucose, and gamma-glutamyl transpeptidase showed significant correlation with LS in univariate analysis, in multivariate analysis only the serum triglyceride level was significantly correlated with LS. CONCLUSION: Steatosis is a relatively common finding in CHB and metabolic host factors rather than viral factors responsible for the presence of steatosis in these patients. PMID- 19190501 TI - Colon cancer in Chile before and after the start of the flour fortification program with folic acid. AB - BACKGROUND: Folate depletion is associated with an increased risk of colorectal carcinogenesis. A temporal association between folic acid fortification of enriched cereal grains and an increase in the incidence of colorectal cancer in the USA and Canada has, however, been recently reported. AIM: To compare the rates of hospital discharges owing to colon cancer in Chile before and after the start of the mandatory flour fortification program with 220 microg of synthetic folic acid/100 g of wheat flour. METHODS: Cancer and cardiovascular hospital discharge rates were compared using rate ratios between two study periods, 1992 1996, before folic acid fortification and 2001-2004, after the flour fortification with folic acid was established in the country. Standard errors of the log rate ratio to derive confidence intervals, and to test the null hypothesis of no difference, were calculated. RESULTS: The highest rate ratio between the two periods was for colon cancer in the group aged 45-64 years (rate ratio: 2.6, confidence interval: 99% 2.93-2.58) and in the 65-79 years (rate ratio: 2.9, confidence interval: 99% 3.25-2.86). CONCLUSION: Our data provide new evidence that a folate fortification program could be associated with an additional risk of colon cancer. PMID- 19190502 TI - Feasibility of a slower lamotrigine titration schedule for bipolar depression: a naturalistic study. AB - The development of a skin rash is often associated with a rapid escalation of lamotrigine dose. We used lamotrigine to treat 259 patients with Diagnostic and Statistical Manual of Mental Disorders, 4th edition, bipolar depression using either the standard titration schedule (n=132) or a slower titration schedule (n=127) and compared the clinical efficacy and safety of both groups. Clinical efficacy of lamotrigine treatment was assessed using changes in the Clinical Global Impression Scale for Bipolar Disorder-Modified scores during the course of the 12-week treatment. A significant reduction of the Clinical Global Impression Scale for Bipolar Disorder-Modified score was observed in both groups and the effect size was large for both groups (standard, 0.75; slower, 0.71). A mixed effect model of repeated measurement revealed an increased rate of improvement in the standard titration group that was significant during the first 5 weeks (P<0.001) but became nonsignificant by the final 7 weeks of treatment. The statistically significant reduction in the development of rashes (P=0.005) was a major advantage for patients in the slower titration group. Although the standard titration schedule generally led to faster recovery from depressive symptoms, the slower titration schedule may be an option for patients with a high risk of rash development. PMID- 19190503 TI - From blue jeans to blue genes. AB - Cutaneous venous anomalies are common. They are blue and vary in size, number, and location and account for most consultations at specialized interdisciplinary clinics for vascular anomalies. Venous lesions are clinically important because they cause pain, dysfunction, destruction of adjacent tissues, and esthetic concern. Only resection and sclerotherapy are helpful, although not always curative. Understanding etiopathogenesis could help design animal models and develop novel therapeutic approaches. John B. Mulliken, MD, envisioned a project to uncover the genetic basis of an inherited form of venous malformation in a large New England family. Recruitment of 2 young fellows resulted in a collaborative project that unraveled the searched-for gene and its mutation. This was an opening for a new era in vascular anomalies. Two blue genes' mutations were discovered, which account for most, if not all, of the inherited forms of venous anomalies, but other genes as well, for rheologically diverse lesions. Differential diagnosis and management has improved, and animal models are being made. This was achieved through the help of Dr Mulliken, who inspired 2 young investigators in blue jeans to find 2 blue genes. PMID- 19190504 TI - Predictors of severity in deformational plagiocephaly. AB - Multiple risk factors for deformational plagiocephaly (DP) have been reported. The purpose of this study was to establish the impact of these variables on the severity of this deformity. A prospective cohort study was performed. Parents completed a standardized questionnaire assessing potential risk factors for DP before assessment. Examination included measurement of transcranial difference (TCD; ie, difference in oblique cranial lengths), evaluation of head tilt, and rotational asymmetry. Pearson correlation coefficient, 1-way analysis of variance, and 2-sample t-test were used to quantify the relationship between identified risk factors and TCD. A total of 434 patients with DP were evaluated. Male-to-female ratio was 2:1; mean gestational age was 36.5 weeks. Deformational plagiocephaly was first appreciated at a mean infant age of 6 weeks. A preexisting diagnosis of torticollis was noted in fewer than 50%. Mean TCD was 11.2 mm. Head tilt was documented in 80% of infants, and mean head rotational asymmetry was 16.4 degrees. Deformational plagiocephaly was more severe in multiple birth pregnancies (P < 0.05), males (P < 0.05), infants with a favorite head position (P < 0.01), preexamination diagnosis of torticollis (P < 0.05), and infants with a head tilt (P < 0.05). Lower gestational age (P < 0.05) and greater head rotational asymmetry (P < 0.0001) were found to correlate with DP severity. This study suggests that the relationship between the severity of DP and certain risk factors can be quantified. The presence and degree of cervical imbalance correlate strongly with deformational cranial asymmetry. PMID- 19190505 TI - Infantile hemangioma: challenges, new insights, and therapeutic promise. AB - Infantile hemangiomas are the most common tumors of infancy. They follow a predictable clinical course, beginning in the first 2 weeks of life with a proliferative phase, dominated by rapidly proliferating endothelial cells, lasting for up to a year. Over the next 7 to 10 years (involuting phase), proliferation is reduced, apoptosis increases, and growth of the lesions slows down and finally stops. The regressed lesion may leave behind flabby fatty tissue in place of an often disfiguring lesion, but many involuted lesions will leave the patient without the need for any corrective surgery. About 20% of hemangiomas are extremely disfiguring and destructive to normal tissue and may even be life threatening. In the last several years, much has been learned about molecular features of hemangioma and hemangioma-derived endothelial cells cultured in vitro, but the specific etiology remains unclear. The abundance of molecular clues from educated guesswork, histology, molecular screening studies, and immunohistochemistry with antibodies against specific proteins have prompted much speculation about the biochemical defect underlying hemangiogenesis, and many pathogenetic mechanisms have been proposed. This review summarizes the current state of knowledge about hemangioma, highlighting the proposed mechanisms that are best supported by the available data and the implications for therapeutic advances. PMID- 19190506 TI - Reconstruction of pediatric cranial base defects: a review of a single microsurgeon's 30-year experience. AB - BACKGROUND: For the past 30 years, microsurgical free tissue transfer has enabled the reconstruction of pediatric cranial base lesions formerly believed to be refractory to surgical therapy. Due to the relative rarity of these oncologic processes and the highly specialized requirements for their treatment, few large scale reviews of microsurgical reconstruction of pediatric cranial base lesions have been published to date. METHODS: A review of all free tissue transfer reconstructive procedures undertaken by a single microsurgeon for pediatric cranial base defects was performed for operations occurring between 1977 and 2007. All procedures were performed at a single institution on patients ranging from infancy to 16 years of age. Data were culled from a combination of patient charts, hospital records, radiographic studies, and clinical photographs. RESULTS: Thirty patient charts were analyzed from the defined 30-year period. The average patient age at the time of diagnosis was 5.3 years (SD = 4.9 years). The most common primary diagnosis was rhabdomyosarcoma (n = 10; 33%). Most patients received chemotherapy (n = 26; 87%) or radiotherapy (n = 16; 53%). Most patients required extirpative hemimaxillectomy or hemimandiblectomy, necessitating reconstruction of intraoral structures in 16 children (53%). Forty free tissue transfers were performed; the most commonly used donor site was the rectus abdominis muscle (n = 19; 48%), followed by the fibula (n = 13; 30%), scapula (n = 5; 13%), latissimus dorsi muscle (n = 2; 5%), and radial forearm (n = 1; 3%). Reconstructive adjuncts included nonvascularized bone grafts (n = 13; 43%) and sural nerve grafts (n = 6; 20%). Short-term perioperative complications were relatively minor; no flap losses were recorded. The most common anticipated long term complications included growth disturbances (n = 10; 33%), resorption of nonvascularized bone grafts (n = 8; 27%), and soft tissue atrophy/contracture (n = 8; 27%). Most patients studied were noted to be surviving (n = 22; 73%), with an average age of 19.2 years (SD, 10.1 years); among those patients who had died (n = 8; 27%), the average age at death was 14.6 years (SD, 6.2 years). The preponderance of patients who had died received their initial surgery and reconstruction during the first 15 years of this study period (n = 7; 88% of subgroup), with death most often due to complications related to extension of the original malignancy through the cranial base. CONCLUSIONS: As advances in oncologic therapy continue to improve survival among pediatric patients experiencing malignancies involving the cranial base, microsurgery simultaneously continues to enable robust options for postextirpative reconstruction and therefore provides a major benefit to the ongoing care of these individuals. PMID- 19190507 TI - Catatonia and autism: a historical review, with implications for electroconvulsive therapy. AB - Current autism research is historically separated from catatonia and other childhood psychotic disorders, although catatonia and autism share several common symptoms (mutism, echolalia, stereotypic speech and repetitive behaviors, posturing, grimacing, rigidity, mannerisms, and purposeless agitation). Electroconvulsive therapy (ECT) effectively treats catatonia and catatonia related conditions of intractable compulsions, tics, and self-injury in people with autism. We assess the incidence of catatonic symptoms in autism, examine emerging ECT indications in people with autism and related developmental disorders, and encourage ethical debate and legal-administrative action to assure equal access to ECT for people with autism. PMID- 19190508 TI - Successful use of donepezil in treatment of cognitive impairment caused by maintenance electroconvulsive therapy: a case report. AB - Cognitive deficits are commonly associated with maintenance electroconvulsive therapy (ECT). Treatment of these cognitive deficits is important but difficult. Cholinergic pathways are involved in these side effects, and donepezil, a cholinesterase inhibitor, may be useful for improvement of cognition. In this case report, we describe for the first time successful use of donepezil in treating cognitive deficits associated with maintenance ECT. The role of cholinesterase inhibitors in the treatment of cognitive deficits caused by ECT needs further studies. PMID- 19190510 TI - Electroconvulsive therapy in the Netherlands: a questionnaire survey on contemporary practice. AB - OBJECTIVE: To investigate contemporary Dutch practice of electroconvulsive therapy (ECT) and adherence to national and international ECT guidelines. METHODS: Among psychiatrists from all Dutch ECT sites in university and general hospitals (n = 24) and psychiatric hospitals (n = 11), a survey was conducted regarding (1) characteristics of practitioners and institutions, (2) clinical practice of ECT, and (3) technical aspects of ECT. Adherence to 16 criteria selected from (inter)national guidelines was scored. RESULTS: Response rate was 94% (all 24 university and general hospitals and 9 of 11 psychiatric hospitals). Most respondents had extensive experience with ECT (median, 10 years; interquartile range, 4-15 years). Annually, approximately 8.5 sessions of ECT per 10,000 inhabitants were administered. In all ECT sites, 24% used exclusively bilateral electrode placement, 26% used ultrabrief pulse width, and 36% used dosage titration methods. Many practitioners had no knowledge of pulse width (42%) or of current characteristics (12%). Of the 16 investigated criteria, 14 were followed in at least 75% of the institutions. CONCLUSIONS: Although still increasing, the use of ECT in the Netherlands remains modest. Electroconvulsive therapy is generally practiced according to (inter)national guidelines. Electroconvulsive therapy is mostly started unilaterally, and generally, age dependent dosage methods are being used. Knowledge on pulse width and current characteristics is limited. The implementation of updated guidelines might offer the opportunity to further improve practice and stimulate availability of ECT. PMID- 19190511 TI - In the beginning... PMID- 19190512 TI - Combination of electroconvulsive therapy with skin graft surgery for a schizophrenic patient with burns. AB - We report on the use of electroconvulsive therapy (ECT) on a patient with schizophrenia immediately after skin graft surgery for extensive burns. The patient was 22 years old and had been burned in more than 33% of his body because of a suicide attempt. After his admission to a burn unit, he continued to make repeated suicide attempts. Electroconvulsive therapy with a muscle relaxant and intubation was administered after scheduled skin graft surgery. His hallucinations diminished after a series of ECT treatments, and his behavior improved. This combination avoids an anesthetic procedure and makes it possible for physicians to administer ECT earlier for burned psychopharmacological treatment-resistant psychiatric patients. The combination treats a patient's physical and mental illnesses simultaneously. PMID- 19190513 TI - Successful electroconvulsive therapy in a patient with intellectual disability and bipolar disorder, with catatonic features misdiagnosed as encephalopathy. AB - We describe the successful use of electroconvulsive therapy (ECT) in a 39-year old woman with a complex clinical picture including bipolar disorder with catatonic features superimposed on a history of multiple congenital anomalies and intellectual and developmental disabilities. At age 34, the patient presented with marked regression and behavioral disturbance diagnosed as encephalopathy. Multiple medical evaluations were inconclusive, and treatment courses with antibiotics and steroids failed. After psychiatric evaluation, her condition was diagnosed as bipolar disorder with catatonic features. She was treated with ECT with dramatic improvement and resumption of her premorbid level of functioning. We discuss the relevant literature on the use of ECT in patients with intellectual disability. PMID- 19190514 TI - UFT plus vinorelbine in advanced non-small cell lung cancer: a phase I and an elderly patient-directed phase II study. AB - PURPOSE: The combination of tegafur-uracil (UFT) with vinorelbine has provided synergistic activity against non-small cell lung cancer (NSCLC) in experimental models. The recommended dose of UFT in combination with vinorelbine in NSCLC was determined in a phase I study. The phase II study evaluated efficacy and tolerability of this combination in elderly patients. METHODS: Vinorelbine was infused on days 1 and 8, and UFT was administered twice daily on days 2 to 6 and days 9 to 13 of a 3-week cycle. UFT and vinorelbine were increased during the phase I study from 400 to 600 mg/d and 20 to 25 mg/m(2), respectively, in 12 patients. In the phase II portion, previously untreated elderly patients were treated with 600 mg/d UFT and 20 mg/m(2) vinorelbine. RESULTS: At the dose level of 600 mg/d UFT and 25 mg/m(2) vinorelbine, dose-limiting toxicity of neutropenia or neutropenic fever was observed in two of three patients, determining the recommended dose of 600 mg/d UFT and 20 mg/m(2) vinorelbine. In 30 evaluable elderly patients of the phase II study, the response rate was 27% (8/30). The median survival and progression-free survival time was 11.8 (range 2.7-34.8) and 5.0 (range 0.5-32.5) months, respectively. Grade 3 or grade 4 neutropenia and grade 3 anemia occurred in 40% and 7% of phase II patients, respectively. Gastrointestinal toxicity was frequent but mild. As the most serious toxicity, pneumonitis was observed in three patients. CONCLUSION: This combination of UFT and vinorelbine is both feasible and active in the treatment of elderly patients with advanced NSCLC. PMID- 19190515 TI - Time to progression as a surrogate marker for overall survival in patients with advanced non-small cell lung cancer. AB - PURPOSE: With the increasing number of active compounds available for advanced non-small cell lung cancer, it is useful to evaluate whether surrogate end points can replace survival in randomized trials for the rapid and efficient assessment of efficacy. We examined the association between differences in overall survival and time to progression (TTP) using a literature survey. METHODS: We used median TTP (MTTP) and median survival time (MST) from 54 phase III trials of first-line chemotherapy involving 23,457 advanced non-small cell lung cancer patients in a multiple linear regression analysis. The MST ratio in each trial was defined as the ratio of MST in the investigational arm to that in the reference arm. The MTTP ratio was defined similarly. RESULTS: The degree of the association between the MST and MTTP ratios was only moderate both in the overall cohort (R(2) = 0.33) and various trial settings (R(2) = 0.16-0.51), although the MTTP ratio was an independent factor influencing the MST ratio in the multiple regression model (p < 0.01). This means that the MTTP ratio could account for less than half of the variance in the MST ratio. CONCLUSIONS: The TTP potentially acts as a surrogate marker, but may not be still a definitive alternative in the first-line setting. PMID- 19190516 TI - Upregulation of hypoxia-inducible factor-1alpha mRNA and its clinical significance in non-small cell lung cancer. AB - BACKGROUND: Hypoxia-inducible factor 1alpha (HIF-1alpha) is a transcription factor that plays an important role in tumor growth by regulating the energy metabolism and angiogenesis. We herein investigated the mRNA expression level of HIF-1alpha in non-small cell lung cancer (NSCLC) tissues to clarify the impact on the clinical aspects of NSCLC patients. EXPERIMENTAL DESIGN: HIF-1alpha mRNA derived from either a tumor or an adjacent lung tissue was quantified using quantitative reverse transcription polymerase chain reaction in 66 patients with NSCLC. The relationship between the mRNA expression level of HIF-1alpha and clinicopathological factors was investigated. RESULTS: The expression level of HIF-1alpha mRNA, which correlated with its protein level, was significantly higher in tumor tissue than in the corresponding nontumor-bearing lung tissue (4.22 x 10(4) +/- 4.99 x 10(4) versus 1.24 x 10(4) +/- 1.15 x 10(4); p < 0.001). The level of HIF-1alpha mRNA showed a significantly positive correlation with the mRNA levels of vascular endothelial growth factor and type II hexokinase in tumors (p < 0.0001 for each). In node-negative patients, high expression levels of HIF-1alpha mRNA in tumors were associated with a poor prognosis (p = 0.0401), but not in the node-positive cases. CONCLUSION: The expression of HIF-1alpha mRNA is associated with disease progression in NSCLC tissues, and is expected as a biomarker or therapeutic target. PMID- 19190517 TI - Establishment of a multi-analyte serum biomarker panel to identify lymph node metastases in non-small cell lung cancer. AB - INTRODUCTION: In non-small cell lung cancer (NSCLC), the presence of locoregional lymph node metastases remains the most important prognostic factor and significantly guides treatment regimens. Unfortunately, currently-available noninvasive staging modalities have limited accuracy. The objective of this study was to create a multianalyte blood test capable of discriminating a patient's true (pathologic) nodal status preoperatively. METHODS: Pretreatment serum specimens collected from 107 NSCLC patients with localized disease were screened with 47 biomarkers implicated in disease presence or progression. Multivariate statistical algorithms were then used to identify the optimal combination of biomarkers for accurately discerning each patient's nodal status. RESULTS: We identified 15 candidate biomarkers that met our criteria for statistical relevance in discerning a patient's preoperative nodal status. A 'random forest' classification algorithm was used with these parameters to define a 6-analyte panel, consisting of macrophage inflammatory protein-1alpha, carcinoembryonic antigen, stem cell factor, tumor necrosis factor-receptor I, interferon-gamma, and tumor necrosis factor-alpha, that was the optimum combination of biomarkers for identifying a patient's pathologic nodal status. A Classification and Regression Tree analysis was then created with this panel that was capable of correctly classifying 88% of the patients tested, relative to the pathologic assessments. This value is in contrast to our observed 85% classification rate using conventional clinical methods. CONCLUSIONS: This study establishes a serum biomarker panel with efficacy in discerning preoperative nodal status. With further validation, this blood test may be useful for assessing nodal status (including occult disease) in NSCLC patients facing tumor resection therapy. PMID- 19190518 TI - Chronic obstructive lung diseases and risk of non-small cell lung cancer in women. AB - INTRODUCTION: The link between lung cancer and chronic obstructive lung diseases (COPD) has not been well studied in women even though lung cancer and COPD account for significant and growing morbidity and mortality among women. METHODS: We evaluated the relationship between COPD and non-small cell lung cancer in a population-based case-control study of women and constructed a time course of chronic lung diseases in relation to onset of lung cancer. Five hundred sixty-two women aged 18 to 74, diagnosed with non-small cell lung cancer and 564 population based controls matched on race and age participated. Multivariable unconditional logistic regression models were used to estimate risk associated with a history of COPD, chronic bronchitis, or emphysema. RESULTS: Lung cancer risk increased significantly for white women with a history of COPD (odds ratios [OR] = 1.85; 95% confidence intervals [CI]: 1.21-2.81), but this was not seen in African American women. Risk associated with a history of chronic bronchitis was strongest when diagnosed at age 25 or earlier (OR = 2.35, 95% CI: 1.17-4.72); emphysema diagnosed within 9 years of lung cancer was also associated with substantial risk (OR = 6.36, 95% CI: 2.36-17.13). Race, pack-years of smoking, exposure to environmental tobacco smoke as an adult, childhood asthma, and exposure to asbestos were associated with a history of COPD among lung cancer cases. CONCLUSIONS: In women, COPD is associated with risk of lung cancer differentially by race. Untangling whether COPD is in the causal pathway or simply shares risk factors will require future studies to focus on specific COPD features, while exploring underlying genetic susceptibility to these diseases. PMID- 19190519 TI - Risk of intracranial hemorrhage and cerebrovascular accidents in non-small cell lung cancer brain metastasis patients. AB - BACKGROUND: Brain metastases confer significant morbidity and a poorer survival in non-small cell lung cancer (NSCLC). Vascular endothelial growth factor targeted antiangiogenic therapies (AAT) have demonstrated benefit for patients with metastatic NSCLC and are expected to directly inhibit the pathophysiology and morbidity of brain metastases, yet patients with brain metastases have been excluded from most clinical trials of AAT for fear of intracranial hemorrhage (ICH). The underlying risk of ICH from NSCLC brain metastases is low, but needs to be quantitated to plan clinical trials of AAT for NSCLC brain metastases. METHODS: Data from MD Anderson Cancer Center Tumor Registry and electronic medical records from January 1998 to March 2006 was interrogated. Two thousand one hundred forty-three patients with metastatic NSCLC registering from January 1998 to September 2005 were followed till March 2006. Seven hundred seventy-six patients with and 1,367 patients without brain metastases were followed till death, date of ICH, or last date of study, whichever occurred first. RESULTS: The incidence of ICH seemed to be higher in those with brain metastasis compared with those without brain metastases, in whom they occurred as result of cerebrovascular accidents. However, the rates of symptomatic ICH were not significantly different. All ICH patients with brain metastasis had received radiation therapy for them and had been free of anticoagulation. Most of the brain metastasis-associated ICH's were asymptomatic, detected during increased radiologic surveillance. The rates of symptomatic ICH, or other cerebrovascular accidents in general were similar and not significantly different between the two groups. CONCLUSIONS: In metastatic NSCLC patients, the incidence of spontaneous ICH appeared to be higher in those with brain metastases compared with those without, but was very low in both groups without a statistically significant difference. These data suggest a minimal risk of clinically significant ICH for NSCLC brain metastasis patients and proposes having more well designed prospective trail to see the role of AAT in this patient population. PMID- 19190520 TI - Cytokeratin expression in malignant melanoma: potential application of in-situ hybridization analysis of mRNA. AB - Occasional reports indicated cytokeratin (CK) protein expression (mainly by immunohistochemistry) in malignant melanoma (MM) and suggested an association with unfavorable clinical parameters. However, the mRNA expression of CK and its clinicopathologic significance in MM has not been specifically evaluated. We investigated the mRNA and protein expression of nine CKs in melanoma cell lines and tissues, in particular the prognostic significance of CK18 mRNA expression. Reverse transcription (RT)-PCR (CK6-10, 14 and 18-20), in-situ hybridization (ISH) (CK18), and western blotting (CK18 and pan-cytokeratin AE1/AE3) were performed on MM cell lines A375, A875, M14, and SK-MEL-1. Eighty MM tissue samples were analyzed by ISH and immunohistochemistry for CK18 expression. The mRNA of CK6-8, 10, 14, 18, and 19 (but not CK9 and 20) was detected in one to four of the melanoma cell lines by RT-PCR. CK18 was detected in all four cell lines by RT-PCR, ISH, and western blotting. CK18 mRNA ISH was positive in three of 30 (10.0%), 10 of 25 (40.0%), and 12 of 25 (48.0%) of primary cutaneous, primary mucosal, and metastatic melanomas, respectively (overall positivity: 25 of 80, 31.3%). CK18 immunostaining was only observed focally in eight of 80 (10.0%) of MM tissue samples, and AE1/AE3 immunostaining was altogether negative. Significantly, CK18 mRNA ISH positivity (but not protein immunohistochemistry) was associated with poorer prognosis by both univariate analysis (P<0.001) and multivariate analysis (relative risk=5.430, 95% confidence interval 2.246-13.128, P<0.001). CK18 mRNA could be identified in one-third of melanoma tissue samples and is an adverse prognostic factor. ISH is superior to immunohistochemistry for analyzing CK18 expression in MM. PMID- 19190521 TI - Multisensory processing and neural oscillatory responses: separation of visuotactile congruency effect and corresponding electroencephalogram activities. AB - By using electroencephalogram, we aimed at identifying the neural oscillations corresponding to two subcomponents of the visuotactile congruency process, facilitation and interference effects. The participants identified the location of the tactile stimulus presented at their index finger or thumb while ignoring simultaneous visual stimuli presented at the same (congruent) or different (incongruent) finger. Significant differences in reaction time were found between the congruent or incongruent condition and the unimodal baseline condition, indicating two different effects of visuotactile interaction. Electroencephalogram results showed significant increases in the gamma-band response in the congruent condition and in the theta-band response in the incongruent condition. These results suggest two systems of multisensory interaction. PMID- 19190522 TI - Distinct neural processes of bodily awareness in crossed fingers illusion. AB - The tactile reassignment process supports the flexible and dynamic changes of body schema in various situations such as those involving tool use. Here, we show that there exist two distinct neural processes in the dynamical reassignment process. One process is involved in identifying the body part where the tactile stimuli are applied, whereas the other is involved in the assignment of the tactile stimuli in the external space including one's body. These processes, combined together, would facilitate the quick and appropriate acquisition of information from the environment, resulting in the speedy spatial perception and execution of motor activities. In addition, we show that the body posture affects the accuracy of tactile localization in the crossed fingers illusion. PMID- 19190523 TI - 5-HT antagonists NAN-190 and SB 269970 block alpha2-adrenoceptors in the guinea pig. AB - Serotonin (5-HT) plays a significant role in the regulation of intestinal secretion of water and electrolytes. The initial aim of this study was to use intracellular recording and specific antagonists to identify roles of 5-HT1A and 5-HT7 receptors of submucosal noncholinergic secretomotor neurons of guinea pig ileum, in vitro. However, it was found that the widely used 5-HT receptor antagonists NAN-190 (5-HT1A) and SB 269970 (5-HT7) both blocked alpha2 adrenoceptors, and hence depressed inhibitory synaptic potentials and hyperpolarizations evoked by noradrenaline, in these neurons. Both compounds enhanced neurally evoked contractions of the guinea pig vas deferens, an effect characteristic of blockade of alpha2-adrenoceptors. These results raise significant concerns about studies using NAN-190 and SB 269970 as specific antagonists of serotonin receptors. PMID- 19190525 TI - Clinical manifestations, etiology, and pathogenesis of the hyper-IgE syndromes. AB - Autosomal dominant Hyper-IgE syndrome (AD-HIES) is a rare primary immunodeficiency characterized by eczema, recurrent skin and lung infections, elevated serum IgE, and various connective tissue, skeletal, and vascular abnormalities. Mutations in signal transducer and activator of transcription 3 (STAT3) have recently been found to account for most cases; however, the pathogenesis of the varied features remains poorly defined. A distinct syndrome, known as autosomal recessive HIES (AR-HIES) manifests as severe eczema, recurrent bacterial and viral skin infections, and sinopulmonary infections. As opposed to STAT3 deficient HIES, AR-HIES lacks the connective tissue and skeletal manifestations but has an increase in neurologic abnormalities. In this review, we discuss the clinical presentations, genetic etiologies, and immunologic abnormalities of these two syndromes. In addition, we discuss animal models of STAT3 deficiency that provide insight into the pathogenesis of HIES. Further understanding of how STAT3 results in the diverse manifestations of HIES will allow us to develop more specific therapies for HIES as well as for many of the manifestations, such as scoliosis, recurrent staphylococcal infections, and eczema, which are common in the general population. PMID- 19190524 TI - Host genetic determinants of human immunodeficiency virus infection and disease progression in children. AB - Increasing data support host genetic factors as an important determinants of human immunodeficiency virus type-1 (HIV-1) susceptibility, mother-to-child transmission (MTCT), and disease progression. Of these genetic mediators, those impacting innate and adaptive immune responses seem to play a critical role in viral infectivity and pathogenesis. During primary infection, CCR5 using virus is predominantly transmitted and polymorphisms that affect the expression of CCR5 alter the risk for MTCT and rate of disease. Chemokines that naturally bind to coreceptors alter infectivity and viral pathogenesis. Additional genes that affect innate immunity including those encoding for MBL2 and those modulating the adaptive immune response including CX3CR1 and human leukocyte antigen types can significantly modify susceptibility and response to HIV-1 infection. As young children develop, the dependence on certain arms of the immune system varies and can alter the effect of genetic variants. Additionally, host genetic factors may alter the response to antiretrovirals. Finally, because HIV-infected children progress more rapidly than adults and have fewer background cofactors, such as drug use and coinfections, the effects of host factors on HIV-1 disease may be more clearly identified. In this review, we summarize available data on the impact of host genetics on MTCT and disease progression of HIV-infected children. PMID- 19190526 TI - Monogenic autoimmune diseases: insights into self-tolerance. AB - Autoimmune diseases affect a significant segment of the population and are typically thought to be multifactorial in etiology. Autoimmune diseases due to single gene defects are rare, but offer an invaluable window into understanding how defects in the immune system can lead to autoimmunity. In this review, we will focus on autoimmune polyendocrinopathy syndrome type 1 and recent advances in our understanding of this disease. We will also discuss two other monogenic autoimmune diseases: immunodysregulation, polyendocrinopathy, and enteropathy, X linked and Autoimmune lymphoproliferative syndrome. Importantly, the knowledge and principles gained from studying these diseases have been applicable to more common autoimmune diseases and have opened the door to better diagnostic and therapeutic modalities. PMID- 19190527 TI - Molecular pathogenesis of Staphylococcus aureus infection. AB - S. aureus has evolved a comprehensive strategy to address the challenges posed by the human immune system. The emergence of community-associated methicillin resistant S. aureus (CA-MRSA) infections in individuals with no predisposing conditions suggests an increased pathogenicity of the bacterium, which may be related to acquisition of novel genetic elements. Remarkably, despite an abundance of research, the underlying cause of the epidemic is not known. Here, the various strategies used by S. aureus to evade obstacles laid out by the human host during colonization and infection were reviewed. The controversies surrounding MRSA research were described, and how acquisition of the novel genes could explain the increased incidence and severity of CA-MRSA diseases was described. PMID- 19190529 TI - Common variable immunodeficiency. AB - Common variable immunodeficiency is the most prevalent clinically significant antibody deficiency at all ages. The disorder is defined principally by characteristic infection susceptibility with hypogammaglobulinemia and impaired specific antibody response. Several recent large registry-based studies have defined distinct phenotypic subtypes. Several studies have also correlated specific immunologic markers with these phenotypes. The biochemical or genetic abnormality in the majority of patients remains unknown. Recently, several molecular genetic lesions have been defined. Among these, mutations of inducible costimulator, and CD19, appear to be disease causing by themselves. These account for about 1% of cases. Other mutations or polymorphisms, such as in the human homolog of Escherichia coli MutS 5 (MSH5), and transmembrane activator and calcium mobilizing ligand interactor, seem to be disease associated in 5-10% of patients, but may require additional immunologic abnormalities for full expression of the phenotype, as unaffected heterozygotes have also been described. PMID- 19190530 TI - Recent advances in primary immunodeficiencies: identification of novel genetic defects and unanticipated phenotypes. AB - Primary immunodeficiencies (PIDs) have traditionally been defined according to their immunologic phenotype. Far from being concluded, the search for human genes that, when mutated, cause PID is actively being pursued. During the last year, four novel genetic defects that cause severe combined immunodeficiency and severe congenital neutropenia have been identified. At the same time, the immunologic definition of primary immunodeficiencies has been expanded by the recognition that genetic defects affecting innate immunity may result in selective predisposition to certain infections, such as mycobacterial disease, herpes simplex encephalitis, and invasive pneumococcal infections. Studies of genetically determined susceptibility to infections have recently shown that immunologic defects may also account for novel infectious phenotypes, such as malaria or leprosy. Finally, a growing body of evidence indicates that primary immunodeficiencies may present with a noninfectious clinical phenotype that may be restricted to single organs, as in the case of atypical hemolytic uremic syndrome or pulmonary alveolar proteinosis. Overall, these achievements highlight the importance of human models, which often differ from the corresponding animal models. PMID- 19190528 TI - Role of Ureaplasma species in neonatal chronic lung disease: epidemiologic and experimental evidence. AB - The contribution of Ureaplasma respiratory tract colonization to the pathogenesis of bronchopulmonary dysplasia in preterm infants has been debated for over 20 y. We review the current understanding of the role of inflammation in altered developmental signaling in the preterm lung and the evidence from human studies and experimental models that Ureaplasma-mediated inflammation produces the BPD phenotype. We propose that Ureaplasma infection initiated in utero and augmented postnatally by exposure to volutrauma and oxygen elicits a sustained, dysregulated inflammatory response in the immature lung that impairs alveolarization, and stimulates myofibroblast proliferation and excessive collagen and elastin deposition. Potential strategies to prevent or ameliorate the effects of Ureaplasma infection in utero and in the preterm lung are discussed. PMID- 19190531 TI - An integrated classification of pediatric inflammatory diseases, based on the concepts of autoinflammation and the immunological disease continuum. AB - Historically, pediatric inflammatory diseases were viewed as autoimmune but developments in genetics of monogenic disease have supported our proposal that "inflammation against self" be viewed as an immunologic disease continuum (IDC), with genetic disorders of adaptive and innate immunity at either end. Innate immune-mediated diseases may be associated with significant tissue destruction without evident adaptive immune responses and are designated as autoinflammatory due to distinct immunopathologic features. However, the majority of pediatric inflammatory disorders are situated along this IDC. Innate immunity has been demonstrated in polygenic disorders, particularly Crohn's disease (CD). A genetic overlap exists between CD and some major histocompatibility complex (MHC) class I associated diseases, including psoriasis; these diseases seem to represent a true intermediate between autoinflammation and autoimmunity. Conversely, classical autoimmune diseases, with autoantibody and MHC class II associations, including celiac disease and rheumatoid arthritis (RA), have adaptive immune genetic associations, including Cytotoxic T-Lymphocyte Antigen-4 (CTLA4) and PTPN22. This proposed classification is clinically relevant, because innate immune-mediated disorders may respond to cytokine antagonism whereas autoimmune-mediated diseases respond better to anti-T and B cell therapies. Furthermore, the etiopathogenesis of poorly defined "autoimmune" diseases, such as juvenile idiopathic arthritis, may be inferred to have substantial innate immune involvement, based on response to IL-1 antagonism. PMID- 19190532 TI - The vitamin D connection to pediatric infections and immune function. AB - Over the past 20 y, a resurgence in vitamin D deficiency and nutritional rickets has been reported throughout the world, including the United States. Inadequate serum vitamin D concentrations have also been associated with complications from other health problems, including tuberculosis, cancer (prostate, breast, and colon), multiple sclerosis, and diabetes. These findings support the concept of vitamin D possessing important pleiotropic actions outside of calcium homeostasis and bone metabolism. In children, an association of nutritional rickets with respiratory compromise has long been recognized. Recent epidemiologic studies clearly demonstrate the link between vitamin D deficiency and the increased incidence of respiratory infections. Further research has also elucidated the contribution of vitamin D in the host defense response to infection. However, the mechanism(s) by which vitamin D levels contribute to pediatric infections and immune function has yet to be determined. This knowledge is particularly relevant and timely, because infants and children seem more susceptible to viral rather than bacterial infections in the face of vitamin D deficiency. The connection among vitamin D, infections, and immune function in the pediatric population indicates a possible role for vitamin D supplementation in potential interventions and adjuvant therapies. PMID- 19190534 TI - Molecular genetics of Kawasaki disease. AB - Kawasaki disease (KD) is a leading cause of acquired cardiac disease of children in the developed countries. The pathogen that triggers this perplexing disease is still unknown after 40 y from the first description. Epidemiologic findings have made us believe that there are considerable genetic components in the etiology and some candidate genetic variations, which confer susceptibility to KD or risk for coronary artery lesions have been identified. However, most of them remain to be definitively confirmed by replication studies with large cohorts. In this article, I review the candidate gene association studies to date. I also introduce our recent findings in genome-wide approach, which revealed the importance of Ca2+/nuclear factor of activated T-cells pathway in the pathogenesis of KD. PMID- 19190535 TI - Human metapneumovirus and human bocavirus in children. AB - Several new viruses have recently been described in children, including human metapneumovirus (hMPV) and human bocavirus (HBoV). hMPV has been established as a common cause of upper and lower respiratory tract infections in children, often second only to respiratory syncytial virus as a cause of bronchiolitis in infants. Diagnostic tools have been developed for the clinician and effective treatment and prevention strategies are being investigated. HBoV was more recently identified. Although it was initially identified in the airway of children, high rates of codetection of other viral pathogens and detection of the virus in the stool have raised questions about the true role of HBoV as a cause of respiratory infections. A focus on epidemiology, pathogenesis, clinical features, and diagnostic techniques for hMPV and HBoV is presented. PMID- 19190533 TI - Biomarkers for infants at risk for necrotizing enterocolitis: clues to prevention? AB - Necrotizing enterocolitis (NEC) is the most common severe gastrointestinal emergency that affects premature newborns. This disease often has a rapid onset with few, if any, antecedent signs that can be used to reliably predict its occurrence. Its rapid onset and progression to death, as well as its severe morbidity when the infant survives, begs for early diagnostic tools that may be used in determining those infants who would be at greatest risk for development of the disease and for whom early preventative measures could be targeted. Although studies have suggested efficacy of several techniques such as breath hydrogen, inflammatory mediators in blood, urine or stool, and genetic markers, these all have drawbacks limiting their use. The application of newly developed "omic" approaches may provide biomarkers for early diagnosis and targeted prevention of this disease. PMID- 19190537 TI - Establishing functional residual capacity at birth: the effect of sustained inflation and positive end-expiratory pressure in a preterm rabbit model. AB - The effect of a 20 s sustained inflation (SI) and positive end-expiratory pressure (PEEP) on functional residual capacity (FRC) formation at birth were investigated. Preterm rabbit pups (28 d) were randomized at birth into four groups (n = 6 for each): 1) SI, PEEP 5 cm H2O, 2) no SI, PEEP 5 cm H2O, 3) no SI + no PEEP, 4) SI + no PEEP. FRC and tidal volume (Vt) were measured by plethysmography and uniformity of lung aeration by phase contrast x-ray imaging. Ventilation with a SI and PEEP uniformly aerated the lung and Vt and FRC were recruited by the first tidal inflation. Ventilation without a SI, with PEEP, gradually recruited Vt and FRC with each inflation but aeration was not uniform. Ventilation without a SI or PEEP, gradually recruited Vt, but no FRC. Ventilation with a SI, without PEEP, uniformly aerated the lung and recruited Vt but no FRC. FRC was greater with SI (p = 0.006) during the first minute, but was larger with PEEP than without PEEP throughout the first 7 min (p < 0.0005). Effects of PEEP and SI were additive. In ventilated preterm rabbits at birth, combining a SI and PEEP improved FRC formation and uniformity of lung aeration, but PEEP had the greatest influence. PMID- 19190536 TI - Developmental changes in brainstem neurons regulating lower airway caliber. AB - Premature infants are at risk for lower airway obstruction; however, maturation of reflex pathways regulating lower airway patency is inadequately studied. We hypothesized that postnatal maturation causes developmental change in brainstem efferent airway-related vagal preganglionic neurons (AVPNs) within the rostral nucleus ambiguus (rNA) that project to the airways and in pulmonary afferent fibers that terminate in the nucleus tractus solitarius (NTS). Ferrets aged 7, 14, 21, and 42 d received intrapulmonary injection of cholera toxin (CT)-beta subunit, a transganglionic retrograde tracer. Five days later, their brainstem was processed for dual immunolabeling of CT-beta and the cholinergic marker, choline acetyl transferase. CT-beta-labeled AVPNs and CT-beta-labeled afferent fiber optical density (OD) were analyzed. There was a significantly higher CT beta-labeled cell number within the rNA at the youngest compared with older ages. All efferent CT-beta-labeled cells expressed choline acetyl transferase. OD of CT beta-labeled afferent fibers was also higher at 7 d compared with 14 d. We conclude that the number of efferent AVPNs and afferent fiber OD both diminish over the second postnatal week. We speculate that exposure to injurious agents in early postnatal life may inhibit natural remodeling and thereby enhance later vulnerability to airway hyperreactivity. PMID- 19190538 TI - Suppression of nonsense mutations in Rett syndrome by aminoglycoside antibiotics. AB - Rett Syndrome (RTT) is caused in more than 60% of cases by nonsense mutations in the MECP2 gene. So far, no curative therapy for RTT has become available. In other genetic disorders, it has been shown that aminoglycosides can cause a read through of nonsense mutations with an efficiency of up to 20%. The aim of this study was to evaluate if this therapeutic concept is applicable to RTT. HeLa cells were transfected with eukaryotic expression vectors carrying mutant alleles of frequently occurring MECP2 nonsense mutations that were N-terminally fused to a FLAG tag. Transfected cells were incubated 24 h in the presence of gentamicin. The expression of full-length protein was analyzed by Western blotting and immunofluorescent cell staining. In the presence of gentamicin a read-through varying between 10 and 21.8% was found, depending on the nucleotide sequence context of the nonsense mutations. The full-length protein was located correctly in the nucleus. We have shown that aminoglycoside-mediated read-through of nonsense mutations in the MECP2 gene can be achieved in vitro with efficiency comparable with that seen in other disorders. PMID- 19190539 TI - Brain-derived neurotrophic factor in infants <32 weeks gestational age: correlation with antenatal factors and postnatal outcomes. AB - Neurotrophins (NTs) play important roles in brain growth and development. Cord blood (CB) brain-derived neurotrophic factor (BDNF) concentrations increase with gestational age but data regarding postnatal changes are limited. We measured BDNF concentrations after birth in 33 preterm infants <32-wk gestation. Serum was collected at birth (CB), at day 2, between day 6 and 10 (D6), at day 30 (D30), and at day 60 (D60). BDNF concentrations fell on D2 (p = 0.03), recovered by D6 (p = 0.10), and continued to rise thereafter at D30 (p = 0.06) and D60 (p = 0.01) compared with CB. CB BDNF concentrations positively correlated with duration of rupture of membranes (r = 0.43, p = 0.04). Antenatal steroids (ANS, p = 0.02), postnatal steroids (PNS, p = 0.04), and retinopathy of prematurity (ROP, p = 0.02) were identified as significant factors in multivariate analyses. The median (25-75th interquartile range) CB BDNF concentrations were higher in infants who received a complete course ANS compared with those who received a partial course [1461 (553-2064) versus 281 (171-536) pg/mL, p = 0.04]. BDNF concentrations negatively correlated with the use of PNS at D30 (r = -0.53, p = 0.002) and at D60 (r = -0.55, p = 0.009). PNS use was associated with reduced concentrations of BDNF at D30 [733 (101-1983) versus 2224 (1677- 4400) pg/mL, p = 0.004] and at D60 [1149 (288-2270) versus 2560 (1337-5166) pg/mL, p = 0.01]. BDNF concentrations on D60 in infants who developed ROP (n = 16) were lower than those who did not develop ROP (n = 7) [1417 (553-2540) versus 3593 (2620-7433) pg/mL, respectively, p = 0.005]. Our data suggests that BDNF concentrations rise beyond the first week of age. BDNF concentrations correlate with factors that influence neurodevelopment outcomes. PMID- 19190540 TI - A pharmacokinetic and dosing study of intravenous insulin-like growth factor-I and IGF-binding protein-3 complex to preterm infants. AB - In preterm infants, low levels of insulin like growth factor 1 (IGF-I) have been associated with impaired growth and retinopathy of prematurity. Our objective was to study safety and pharmacokinetics of i.v. administered rhIGF-I with its binding protein 3 (rhIGFBP-3) to preterm infants. At 3 d chronological age, an i.v. 3 h infusion of rhIGF-I/rhIGFBP-3 was administered followed by serial measurements of IGF-I and IGFBP-3. Infants were evaluated for physiologic safety measurements. The individual dose of rhIGF-I ranged from 1 to 12 microg/kg. The study was conducted at Queen Silvia Children's Hospital, Gothenburg, Sweden, between January and November 2007. Five patients (3 F) with mean (range) post menstrual age 27 wk (26-29) and birth weight 1022 g (810-1310) participated. IGF I and IGFBP-3 levels before infusion were median (range) 18 (12-28) and 771 (651 1047) ng/mL, respectively. Immediately after study drug infusion, serum IGF-I and IGFBP-3 levels were 38 (25-59) and 838 (754-1182) ng/mL, respectively. Median (range) half-life for IGF-I and IGFBP-3 was 0.79 (0.59-1.42) and 0.87 (0.85-0.94) hours, respectively. Blood glucose, insulin, sodium, potassium, and physiologic safety measures were within normal ranges. The rhIGF-I/rhIGFBP-3 equimolar proportion was effective in increasing serum IGF-I levels and administration under these study conditions was safe and well tolerated. PMID- 19190541 TI - Whole body magnetic resonance imaging of healthy newborn infants demonstrates increased central adiposity in Asian Indians. AB - Abdominal adiposity and metabolic ill health in Asian Indians are a growing public health concern. Causal pathways are unknown. Preventive measures in adults have had limited success. The aim of this observational case-control study was to compare adipose tissue partitioning in 69 healthy full term Asian Indian and white European newborns born in Pune, India and London, UK, respectively. The main outcome measures were total and regional adipose tissue content measured by whole body magnetic resonance imaging. Although smaller in weight (95% CI for difference -0.757 to -0.385 kg, p < 0.001), head circumference (-2.15 to -0.9 cm, p < 0.001), and length (-2.9 to -1.1 cm p < 0.001), the Asian Indian neonates had significantly greater absolute adiposity in all three abdominal compartments, internal (visceral) (0.012-0.023 L, p < 0.001), deep s.c. (0.003-0.017 L, p = 0.006) and superficial s.c. (0.006-0.043 L, p = 0.011) and a significant reduction in nonabdominal superficial s.c. adipose tissue (-0.184 to -0.029 L, p = 0.008) in comparison to the white European babies despite similar whole body adipose tissue content (-0.175 to 0.034 L, p = 0.2). We conclude that differences in adipose tissue partitioning exist at birth. Investigative, screening, and preventive measures must involve maternal health, intrauterine life, and infancy. PMID- 19190542 TI - Oscillations in cardiovascular function during acute hypoxia in the newborn piglet are associated with less neurological damage and occur more frequently in females. AB - The function of the cardiovascular system during hypoxia is an important determinant of neurologic outcome. Oscillations in blood pressure, particularly type-3 waves with a duration of 10 to 160 s, have been shown to occur in the presence of hypoxia in the neonatal rat. The aim of this study was to determine the characteristics and occurrence of type-3 waves in the neonatal piglet and any relationship to neurologic damage after acute global hypoxia. Hypoxia/hypercarbia was induced in 32 anesthetized piglets by reducing the fraction of inspired oxygen to 0.1 and the ventilation rate from 30 to 10 breaths per minute for 45 min. The degree of neurologic damage was assessed using both physiologic (EEG amplitude and cerebral impedance at 6 h posthypoxia) and structural (microtubule associated protein-2 immunohistochemistry) markers. Type-3 waves in cardiovascular function occurred in 56% of animals. An oscillating pattern was significantly associated with less neurologic damage (p = 0.01) and a lower duration of hypotension during hypoxia (p = 0.02), and occurred more frequently in females (p = 0.024). PMID- 19190543 TI - Microarray analysis of high-dose recombinant erythropoietin treatment of unilateral brain injury in neonatal mouse hippocampus. AB - Recombinant human erythropoietin (rEpo) is neuroprotective in neonatal models of brain injury. Proposed mechanisms of neuroprotection include activation of gene pathways that decrease oxidative injury, inflammation, and apoptosis, while increasing vasculogenesis and neurogenesis. To determine the effects of rEpo on gene expression in 10-d-old BALB-c mice with unilateral brain injury, we compared microarrays from the hippocampi of brain-injured pups treated with saline or rEpo to similarly treated sham animals. Total RNA was extracted 24 h after brain injury and analyzed using Affymetrix GeneChip Mouse Exon 1.0 ST Arrays. We identified sex-specific differences in hippocampal gene expression after brain injury and after high-dose rEpo treatment using single-gene and gene set analysis. Although high-dose rEpo had minimal effects on hippocampal gene expression in shams, at 24-h post brain injury, high-dose rEpo treatment significantly decreased the proinflammatory and antiapoptotic response noted in saline-treated brain-injured comparison animals. PMID- 19190544 TI - Long-term reduction of hippocampal brain-derived neurotrophic factor activity after fetal-neonatal iron deficiency in adult rats. AB - Fetal-neonatal iron deficiency acutely alters hippocampal biochemistry, neural morphology, and electrophysiology accompanied by a downregulation of brain derived neurotrophic factor (BDNF). These changes provide a cellular and molecular basis for observed short-term learning and memory impairments. However, the etiology of residual, long-term hippocampal neurotransmission abnormalities and learning impairments after treatment remain unclear. Because BDNF modulates learning and memory, we assessed its expression in 65-d-old formerly iron deficient (FID) male rats that had been iron deficient during the fetal-neonatal period and treated with iron since postnatal day 7. BDNF-III and -IV mRNAs and BDNF protein expression remained down-regulated in FID rats when compared with the always iron-sufficient rats. Expressions of BDNF activity-dependent downstream targets (3-hydroxy-3-methylglutaryl CoA reductase and immediate early genes c-fos, early growth response gene 1 and 2) were reduced in FID rats. In turn, hippocampal expressions of direct targets of early-growth response genes, including hypoxia-inducible factor 1, dual-specificity phosphatase 4, IGF 2, and myelin basic protein were also diminished in FID rats. Collectively, fetal neonatal iron deficiency lowers hippocampal BDNF expression and function beyond the period of iron deficiency. These findings may underlie the persistence of learning deficits seen after fetal-neonatal iron deficiency. PMID- 19190545 TI - Infantile spasms and Down syndrome: a new animal model. AB - Infantile spasms is a catastrophic childhood seizure disorder for which few animal models exist. Children with Down syndrome are highly susceptible to infantile spasms. The Ts65Dn mouse is a valid model for Down syndrome; therefore, we tested the hypothesis that the Ts65Dn mouse represents a substrate for an animal model of infantile spasms. The baseline of naive Ts65Dn mice showed spontaneous spike-and-wave discharges, a pattern that worsened with baclofen and gamma-butyrolactone, which induced acute epileptic extensor spasms (AEES) associated with epileptiform polyspike bursts and an electrodecremental response on the EEG. GABABR-agonist-induced AEES were significantly reduced with vigabatrin, rodent ACTH fragment, valproic acid, ethosuximide, and CGP 35348. Porcine ACTH had no effect. GABABR protein expression was significantly increased in the thalamus and medulla oblongata of Ts65D mice in comparison with wild-type controls. The GABABR agonist-treated Ts65Dn mouse shows the unique clinical, electrographic, and pharmacologic signature of infantile spasms and represents a valid, acute model of this disorder. GABABR-mediated mechanisms may contribute to the increased susceptibility of children with Down syndrome to infantile spasms. PMID- 19190546 TI - Development of upper body coordination during sitting in typically developing infants. AB - Our goal was to determine how the actions of the thorax and the pelvis are organized and coordinated to achieve independent sitting posture in typically developing infants. The participants were 10 typically developing infants who were evaluated longitudinally from first onset of sitting until sitting independence. Each infant underwent nine testing sessions. The first session included motor evaluation with the Peabody test. The other eight sessions occurred over a period of 4 mo where sitting behavior was evaluated by angular kinematics of the thorax and the pelvis. A physical therapist evaluated sitting behavior in each session and categorized it according to five stages. The phasing relationship of the thorax and the pelvis was calculated and evaluated longitudinally using a one-way analysis of variance. With development, the infants progressed from an in-phase (moving in the same direction) to an out-of phase (moving in an opposite direction) coordinative relationship between the thorax and the pelvis segments. This change was significant for both sagittal and frontal planes of motion. Clinically, this relationship is important because it provides a method to quantify infant sitting postural development, and can be used to assess efficacy of early interventions for pediatric populations with developmental motor delays. PMID- 19190547 TI - Intravitreal ranibizumab, intravitreal ranibizumab with PDT, and intravitreal triamcinolone with PDT for the treatment of retinal angiomatous proliferation: a prospective study. AB - PURPOSE: To compare 1) intravitreal ranibizumab, 2) intravitreal ranibizumab plus photodynamic therapy (PDT), and 3) intravitreal triamcinolone plus PDT in retinal angiomatous proliferation. METHODS: In this prospective study, 37 eyes of 37 patients with retinal angiomatous proliferation were randomly assigned in 1 of the 3 groups. The patients in Group 1 (n = 13) received 3 monthly injections of 0.5 mg of ranibizumab. The patients in Group 2 (n = 13) received one session of PDT and 3 monthly intravitreal injections of 0.5 mg ranibizumab and the patients in Group 3 (n = 11) received one session of PDT and 1 injection of 4 mg triamcinolone. Retreatment, with the same therapeutic scheme in each group, was considered in case of persistence or recurrence of subretinal fluid or intraretinal fluid. RESULTS: All the patients completed at least 6 months of follow-up. A total of 61.53% patients in Group 1, 76.92% in Group 2, and all in Group 3 had the same or better visual acuity at the end of the follow-up (P = 0.0232). The mean central retinal thickness reduction in Group 1 was 32.23 mum (P = 0.548), in Group 2 20.31 mum (P = 0.042), and in Group 3 73.92 mum (P < 0.0001). Also, the patients in Group 3 received on average the lowest number of injections (P < 0.0001). CONCLUSION: All groups resulted in stabilization of the disease, while a significant trend towards better visual acuity and anatomic restoration of the affected area was observed in the intravitreal triamcinolone & PDT group. PMID- 19190548 TI - Dizziness: when is it vascular? PMID- 19190549 TI - Is total abolishment of great saphenous reflux in the invasive treatment of superficial chronic venous insufficiency always necessary? AB - Chronic venous insufficiency ranges from mild telangiectasias to skin ulceration with poor prognosis regarding healing and quality of life. Varicose veins are the most frequent clinical presentation, affecting 75% of such patients and 71% are due to primary reflux of great saphenous vein, which is the most compromised vein in chronic venous insufficiency. However, about 75% of these veins are not dilated. The standard treatment has been stripping of the saphenous vein, because it has 85% of good results at long term. However, saphenous vein is the main arterial substitute and should be spared whenever possible. The development of non-invasive diagnostic methods showed that hemodynamic worsening correlates with clinical severity and that the majority of patients did not have a dilated saphenous vein. Thus, several selective operations proposed to spare the saphenous vein have reported good results. Minimally invasive techniques (eco guided foam, radiofrequency and laser) have also emerged aiming to obliterate the vein and abolishing reflux and have also reported good results, but they do not spare the vessel. Measurement of saphenous diameter has been shown to correlate with clinical and hemodynamic worsening, thus allowing planning the invasive treatment of chronic venous insufficiency. Dilated diameters (>7.2 mm) correlate with severe disease and poor prognosis, being an indication for total abolishment of saphenous vein reflux. All other presentations must be individualized, sparing saphenous vein whenever possible and and a standardized approach is not indicated for all patients. PMID- 19190550 TI - Prognostic role of endothelial dysfunction and carotid intima-media thickness in patients undergoing coronary stent implantation. AB - AIM: Impaired endothelial function and increased carotid intima-media thickness are key events in the atherosclerotic process and predict future cardiovascular events in subjects with and without coronary artery disease. The purpose of this study was to investigate whether the vasodilator response to increased flow in the brachial artery and the presence of carotid lesions may have a prognostic significance for in-stent restenosis in patients undergoing coronary angioplasty. METHODS: The study population included 58 patients undergoing percutaneous coronary intervention (PCI) with stenting and at least 10 months of follow-up. All patients underwent ultrasound detection of brachial artery reactivity 30 days after PCI. Flow mediated dilatation (FMD) was investigated after 5 minutes of occlusion of the artery and nitroglycerin mediated dilation (NMD) was investigated after administration of sublingual nitrates. Vascular echography was performed to measure intima media thickness (IMT) of carotid arteries. At baseline we evaluated all the established traditional cardiovascular risk factors. We also subdivided our study cohort according to values of FMD in patients with FMD above and patients below the median value. RESULTS: Patients with FMD above the median value showed higher prevalence of hypertension (P=0.002), diabetes (P=0.02) and carotid IMT (P=0.006) than those below the median. Brachial FMD was inversely correlated (P=0.001) to carotid IMT. At the end of follow-up clinical events occurred in nine patients. In a multivariate analysis, including all the variables evaluated at baseline, carotid IMT (P=0.02), level of glycemia (P=0.001), a lower FMD (P=0.005) and presence of carotid plaque remained the only variables predictive of restenosis. CONCLUSIONS: Evaluation of FMD and carotid IMT may provide important prognostic information in patients undergoing PCI. PMID- 19190551 TI - Prevalence of peripheral artery disease and its associated risk factors in Spain: The ESTIME Study. AB - AIM: Several studies have demonstrated that patients with peripheral arterial disease (PAD), are at an increased risk of morbidity and mortality compared with those without PAD. However, few population-based studies have addressed the prevalence of PAD and intermittent claudication (IC). We assessed the prevalence of and the factors associated with PAD and IC in the Spanish population. METHODS: A cross sectional study with 1324 participants aged 55 to 84 years randomly selected from the census was conducted in 12 Spanish regions. The presence of PAD and IC was determined by an ankle-brachial index (ABI) <0.90 in either leg and by means of the Edinburgh questionnaire, respectively, fulfilled together with a detailed past history. All participants had blood pressure, body mass index, glycemia, and lipid profile measured. RESULTS: The response rate was 63.9% (846/1 324). ABI prevalence of PAD was 8.03% The prevalence of symptoms of definite or atypical IC was 6%. Subjects with an ABI <0.9 were more likely to be older, men, diabetics, current smokers, with coronary heart disease, with higher systolic pressure and with higher triglyceride levels than participants with ABI 0.9. CONCLUSIONS: ESTIME study confirms the high prevalence of asymptomatic PAD, and its relation with typical cardiovascular risk factors. ABI provides early diagnosis before claudication symptoms in a high proportion of patients. ABI could contribute to developing early prevention programmes. PMID- 19190552 TI - Vacuum assisted wound closure in patients with lower extremity arterial disease. The experience from two tertiary referral-centres. AB - AIM: The purpose of this investigation was to analyze predictors for wound healing, amputation and mortality after vacuum assisted closure (VAC) therapy of wounds in the lower limb in patients with arterial disease. METHODS: One hundred and twenty one wounds were treated and followed for 12 months at two vascular centres in Uppsala and Malmo, Sweden. VAC therapy was applied in the wound at a topical negative pressure of 125 mmHg. RESULTS: Median age of the patients was 74 years and critical lower limb ischemia was present in 87% of the patients at admission. Intestinal flora was cultivated in 74% of the wounds. VAC associated bleeding occurred in four patients. Complete wound healing was achieved in 66%. Deep groin infections were associated with synthetic graft infection (P<0.001), treatment outside hospital (P<0.001), faster healing (P<0.01) and lower amputation rate (P<0.005). Diabetes mellitus (OR 2.7; [95% CI 1.2-6.2]) and foot wound (OR 3.0; [95% CI 1.2-7.4]) were independent predictors for amputation. The absence of complete wound healing was the strongest factor for both amputation (P<0.001) and death (P<0.001). CONCLUSIONS: VAC therapy of complex wounds in the lower limbs in patients with vascular disease was associated with high healing rates. Non-healed wounds after VAC therapy were predictors for amputation and death. PMID- 19190553 TI - Does laser power influence the results of endovenous laser ablation (EVLA) of incompetent saphenous veins with the 1 470-nm diode laser? A prospective randomized study comparing 15 and 25 W. AB - AIM: Major side effects after endovenous laser ablation (EVLA) are pain and bruising. The aim of this study was to compare outcome and side effects after EVLA of incompetent great saphenous veins (GSV) with a 1 470 nm diode laser (Ceralas E, biolitec) using a power of 15 or 25 W. METHODS: Between 28 November 2007 and 15 January 2008, 40 consecutive patients (40 legs) with an incompetent GSV were treated by EVLA. The patients were randomized in two groups. In Group A (20 patients) was used a 15-W-power laser and in Group B (20 patients) a 25-W power laser was used. All patients were re-examined after 1, 10 and 30 days clinically and by Duplex for complications and occlusion in the treated vein segment in a standardized way. RESULTS: There was no significant difference concerning gender, age, C of CEAP, body mass index or diameter of the treated vein. In Group A a mean of 465 mL tumescence fluid (TF) was used and in Group B TF was of 433 mL. In Group A the mean linear endovenous energy density (LEED) was 109.7 J/cm and in Group B 132.6 J/cm. The subgroup Bsub included 16 patients of Group B with a comparable mean LEED of 115.8 J/cm. In all groups occlusion of the treated veins was achieved for all patients. The diameter of the GSV reduced at 3 cm below the sapheno-femoral junction from 1.1 to 0.6 cm, 1.0 to 0.6 cm and 0.9 to 0.6 cm respectively in the three groups. The modified CEAP clinical score improved significantly in all groups. In Group A patients have been administered analgesic tablets for a shorter period. There was also a trend to less postinterventional pain and analgesic use in Group A, but it was not significant. Ecchymosis was rare in both groups (8 in Group A, 7 in Group B). CONCLUSIONS: In this prospective randomized comparative study the power of the laser did not influence the occlusion rate when a high LEED with comparable values was used. In both groups pain and ecchymoses were less frequent in this study with a 1 470 nm diode laser than reported in studies with 810-980 nm systems. A lower power level significantly reduced use of analgesic tablets. PMID- 19190554 TI - Short-term prediction of major lower-limb amputation based on clinical indicators on admission: a single institutional experience in a developing country. AB - AIM: The aim of this study was to analyze the possibility of predicting short term major lower-limb amputation (SMLA) in patients with vascular diagnoses, based only on clinical variables measured on admission. METHODS: A longitudinal, retrospective cohort study of patients with peripheral vascular diagnoses admitted at our Institute was carried out. A stratified sample of 463 patients admitted during 1997, 2000 and 2003, was studied. Logistic regression was used to identify significant predictors of amputation; twelve clinical variables measured on admission were considered as potential predictors. RESULTS: Of the 463 patients, 93 (20%) were amputated. Significant predictors of amputation identified by the logistic regression analysis were ''type of lesion'' (none; neuro-infectious; ischemic; mixture), ''initial diagnosis'' (phlebolymphopathies, acute arterial insufficiency, chronic arterial insufficiency, diabetic foot, others), ''plantar region lesion'' (no; yes), ''diabetes'' (no; yes), ''number of toes affected'' (none; 1-2; 3 or more), and ''area of leg affected'' (none; lower third; + lower third). More than 80% of patients were correctly classified with the final model: sensitivity was 42% and specificity 96%. CONCLUSIONS: It seems that SMLA in patients with vascular diagnoses can be predicted reasonably well using as predictors only clinical variables measured on admission. This is a potentially useful result for Angiology Services located in developing/poor communities. The amputation probability for each patient obtained from the logistic regression model can be used in several ways: 1) the medical care of patients can be customized so that the amputation rate of the whole Service can be reduced, and 2) the amputation probability of the statistical model can be used as an estimation of the severity of the disease in each patient, which in turn can be used to standardize the amputation rates computed on different years; this would allow a better assessment of the Institutional performance over time. PMID- 19190555 TI - Quantification of plasminogen activator inhibitor type 1 in the aortic wall. AB - AIM: Plasminogen activator inhibitor type 1 (PAI-1) plays a key role in regulation of fibrinolytic system, cell-associated proteolysis and migration of smooth muscle cells (SMC). This study is focused on the types of PAI-1 expressing cells, quantification of PAI-1 expression in the walls of aneurysmatic abdominal aortas (AAA) and correlation between histological and clinical findings. METHODS: A group of nine patients who underwent surgery for AAA: asymptomatic (aAAA), symptomatic (sAAA) and ruptured (rAAA) and one control specimen (CA) were included in the study. Samples underwent histological processing and immunohistochemistry in comparison with in situ hybridisation. In order to assess the PAI-1 area fraction in histological sections through the aortic wall the Line System module of Ellipse software was used. PAI-1 expressing cells were measured in CA and AAA: endothelium, SMC, and foam cells. Photomicrographs with a total area of 0.7 mm(2) for each specimen were analysed by two independent observers. Mean values of PAI-1 positive components per section area were calculated as average values. RESULTS: The results of both observers are as follows: 28.6% in CA; 18.1% in aAAA; 10.9% in sAAA; 11.0% in rAAA. During the progression of AAA, the SMC (PAI-1 expression was found mainly in them) became less abundant in agreement with the values of PAI-1 area fraction. In rAAA immunohistochemistry detected PAI-1 in necrotic centres of atheromathous plaques. CONCLUSIONS: AAA may be evaluated as the result of gradual changes in regulation of fibrinolysis that is observed as redistribution of cells expressing PAI-1. The area fraction of PAI 1 positive components correlates with clinical classification of AAA. PMID- 19190556 TI - Lack of major gender-related differences in patients undergoing elective endovascular or surgical repair of abdominal aortic aneurysm. AB - AIM: The aim of this single center retrospective study was to determine gender related differences in patients undergoing repair of aortic aneurysm. METHODS: A total of 296 consecutive patients with an abdominal aneurysm undergoing elective endovascular or surgical repair was included in the study. Among these, 24 (8.1%) were females and 272 (91.9%) were males. Demographic and clinical characteristics as well as laboratory values in terms of any potential gender-specific differences were compared. RESULTS: Females were significantly older at the time of repair than males. No gender-related differences were found analysing major clinical and laboratory parameters. CONCLUSIONS: The clinical and laboratory profile with which males and females present at the time of elective endovascular or surgical repair for aortic aneurysm is very similar. It does not reveal any potential gender-specific risk constellation. It eventually remains unclear why the prevalence of aortic aneurysm is higher in male than in females. PMID- 19190557 TI - Fate of left kidney after left renal vein division during management of aortic occlusive disease. AB - AIM: Left renal vein division (LRVD) has been used as a technical aid to gain exposure to the peri-renal aorta and to control bleeding in abdominal aortic operations. A few retrospective series studied the effects of such division on the overall renal function with contradicting results. The aim of this study was to assess the effect of such division on the left kidney during the management of aortic occlusive disease. METHODS: A prospective study was undertaken on all patients that had abdominal aortic bypass surgery for juxta-renal aortic occlusive disease scheduled between October 2003 and September 2007. Renal function was assessed by measuring serum creatinine (mg/L+/-SD) and creatinine clearance (mL/min+/-SD) preoperatively and postoperatively up to 14 days in patients who had LRVD and patients who had left renal vein intact (LRVI) which served as control. Left kidney was assessed preoperatively by performing abdominal computed tomography angiography (CTA) and postoperatively with either ultrasonography (US) or CTA depending on the serum creatinine level. RESULTS: Thirty-two patients were included in this study. Six patients had LRVD. There was no statistically significant difference between the LRVD and LRVI groups as regard mean serum creatinine, preoperatively (LRVD 1.05+/-0.31 vs LRVI 1.08+/ 0.21,P=0.38) and XIV day postoperatively (LRVD 1.15+/-0.31 vs LRVI 1.09+/-0.24, P=0.34), and mean creatinine clearance, preoperatively (LRVD 94+/-17.4 vs LRVI 97.9+/-11.1, P=0.31) and XIV day postoperatively (LRVD 88.8+/-13.1 vs LRVI 94.3+/ 6.7 p=0.11). CTA and US showed postoperative early diffuse swelling and congestion of the left kidney in 4 cases that had LRVD, which reverted to normal at XIV day postoperatively. CONCLUSIONS: Selective LRVD during aortic occlusive surgery does not compromise the left kidney. PMID- 19190558 TI - Occurrence and management of chronic venous disease in primary health care in Spain. A comparison of DETECT-2006 with DETECT-2000. AB - AIM: The aim of this study was to investigate the prevalence and initial treatment of chronic venous disease (CVD) in the Spanish primary health care system during 2006 and to compare the results with those obtained in 2000. METHODS: The survey involved 1 118 general practitioners co-ordinated by 37 specialists in angiology and vascular surgery, assessing 15 consecutive patients, each attending the clinics between 29 May and 2 June 2006. RESULTS: Of the16 186 patients reported, 82% had CVD risk factors. When asked about CVD signs or symptoms 11 277 patients (69.7%) mentioned some kind of clinical manifestation/sign compatible with the disease. Diagnosis was established in accordance with the clinical section of the CEAP classification, 38% of the total being classified as C2-C6. Sixty-two percent of the symptomatic patients had received prior treatment (vs 24.8% in year 2000), and following the survey 88% of the patients diagnosed with CVD were treated (vs 62% in 2000). CONCLUSIONS: The results for 2006 confirm the high prevalence of CVD in primary health care in Spain. In comparison with the results for 2000, an improvement in the patterns of medical advice use, treatment and prescriptions are observed. Despite these findings, specific training programs in primary health care regarding the diagnosis and treatment of CVD continue to be necessary. PMID- 19190559 TI - Isolated distal deep vein thrombosis: efficacy and safety of a protocol of treatment. Treatment of Isolated Calf Thrombosis (TICT) Study. AB - AIM: The optimal treatment of isolated distal deep vein thrombosis (ID-DVT) is still controversial. A complete anticoagulation as soon as the diagnosis is made is recommended by some authors. Alternatively, other authors suggest to perform serial ultrasonography assessments to detect the possible extension of DVT towards proximal veins. Only in this case the treatment should be initiated. Furthermore, the optimal duration of treatment is far from established. The Treatment of Isolated Calf Thrombosis (TICT) study was set up to assess the efficacy and safety of a particular treatment regimen of ID-DVT based on low molecular weight heparins (LMWH). METHODS: The drug treatment consisted of a twice-daily subcutaneous administration of a full dose of weight-adjusted LMWH for one week, followed by a half dose of LMWH administered once-daily for another three weeks. At the end of the four-week period of treatment, a colour-coded Doppler ultrasonography (CCDU) assessment was scheduled and after three months a follow-up visit was performed. If a patient was unable to attend the visit, he was contacted by a phone-call to assess if any adverse events occurred. The study enrolled 192 outpatients with ID-DVT confirmed by CCDU. Twenty-one out of 192 patients (10.9%) were excluded for violation of protocol. Thus 171 (39.9% men, mean age of 60.45 years ) were eligible and were included in the study. Sixty-one patients (36.6%) presented an unprovoked ID-DVT. RESULTS: Events during the period of treatment (4 weeks). Ten out of 171 patients (5.8%) had complications: five patients showed an extension proximal to the knee (2.9%) all with an unprovoked ID-DVT; two showed an extension of thrombus within the distal veins. Three patients (1.7%) suffered from minor bleeding; there was no major bleeding. Further events during three months of observation occurred. Five patients had thrombus recurrences: four patients showed a proximal DVT (3 with a previous unprovoked ID-DVT, 1 with a previous ID-DVT secondary to a traumatic leg fracture, with persistent difficulty of deambulation); one, with a previous secondary thrombosis, showed a ID-DVT. CONCLUSIONS: In our study only 2.9% of patients with ID-DVT showed a progression of thrombosis to proximal deep veins; the majority of thrombus progression, during the treatment period, was observed in patients with unprovoked ID-DVT. Our results support the usefulness of a prolonged treatment in unprovoked ID-DVT. PMID- 19190560 TI - Does a dedicated nurse practitioner improve thromboprophylaxis use in acutely ill medical patients in Australia? The methodology for a multicentre VTE Task Force Audit. AB - AIM: To determine the extent to which a multifaceted venous thromboembolism (VTE) prophylaxis program, coordinated by a dedicated nurse practitioner, improves the level of appropriate prophylaxis in patients hospitalised with an acute medical illness. METHODS: A multicentre clinical audit was conducted in 16 hospitals across Australia. Approximately 9600 acutely ill medical patients over 18 years of age hospitalised for at least 3 days. A 4-month programme implemented by a VTE Nurse Educator to raise awareness of the risk of VTE, the importance of risk assessment and the appropriate prophylactic management of high-risk medical patients with local VTE prophylaxis audit result feedback. RESULTS: The effect of this programme on the proportion of high-risk medical patients receiving appropriate thromboprophylaxis according to current guidelines. CONCLUSIONS: The VTE Task Force Audit will be the first multicentre clinical audit in Australia to evaluate thromboprophylaxis use in acutely ill medical patients and the effects of employing a nurse educator. Based on published results from clinical audits conducted overseas, it is expected that thromboprophylaxis will be underutilised in these patients. It is hypothesised that an active multifaceted programme will improve the rate of thromboprophylaxis among eligible medical patients through the effective implementation of evidence-based guidelines. PMID- 19190561 TI - Debranching of the aorta produces thrombosis of the false lumen in selected cases of chronic type A aortic dissection. AB - Patent false lumen of the descending aorta is related to poor prognosis in patients who undergo surgery for acute type A aortic dissection. We describe a patient with chronic type A aortic dissection and a patent dilated false lumen that was successfully thrombosed following an ilio-renal bypass. In patients with chronic aortic dissection and a single intimal tear on contrast spiral CT, debranching of arteries of the infra-diaphragmatic aorta supplied by the false lumen and reimplantation of these vessels may produce complete thrombosis of the false lumen. This outcome can spare such patients from extensive aortic replacement or endografts. PMID- 19190562 TI - Skin necrosis during oral anticoagulant long-term treatment: an atypical side. AB - The effectiveness of anticoagulant therapy for venous thromboembolism, with regards to both acute phase and long term prophylaxis, in patients with recurrent deep venous thrombosis (DVT) and persistence of risk factors, has been confirmed by many studies. However, it is not free of complications such as hemorrhage or, more rarely, skin necrosis. The patient, observed by us since 1994, was treated with oral vitamin K antagonists: he was affected by post-thrombotic syndrome and deficiency of congenital procoagulant factors (factor II heterozygote and MTHFR positive heterozygote) and secondary deficiency of procoagulant factors due to the consumption of protein C, with appearance of skin necrosis that occurred after many years of oral anticoagulant treatment. The change of therapy from oral anticoagulant to low molecular weight heparin and the use of local dressing, led to the resolution of the clinical symptoms and on to healing. PMID- 19190563 TI - A liberal preoperative fasting regimen improves patient comfort and satisfaction with anesthesia care in day-stay minor surgery. AB - BACKGROUND: The aim of this study was to evaluate whether a single preoperative limited oral intake of a carbohydrate drink could improve perioperative patient comfort and satisfaction with anesthesia care in elective day-stay ophthalmologic surgery. METHODS: A single-center, prospective, randomized clinical trial was conducted in a university hospital. The study included ASA I-III patients undergoing ophthalmologic surgery. Patients undergoing both general anesthesia and local anesthesia were included in the study. The control group fasted in accordance to nil per os after midnight, while patients in the experimental group received 200 mL of a carbohydrate drink 2 h before the operation. Both groups were allowed to drink and eat until midnight ad libitum. Patient characteristics, subjective perceptions, taste of the drink, and satisfaction with anesthesia care were ascertained using a questionnaire administered three times: after the anesthesiologist's visit, before surgery and before discharge from the ward to assess patient comfort. An analysis of variance and the Mann-Whitney U-test were used for statistical analysis. RESULTS: A total of 123 patients were included and 109 patients were randomly assigned to one of two preoperative fasting regimens. Patients drinking 200 mL 2 h before surgery were not as hungry (P<0.05), not as thirsty preoperatively (P<0.001) and not as thirsty after surgery (P<0.05), resulting in increased postoperative satisfaction with anesthesia care (P<0.05). CONCLUSION: Standardized limited oral preoperative fluid intake increases patient comfort and satisfaction with anesthesia care and should be a part of modern day stay ophthalmologic surgery. PMID- 19190564 TI - Appendectomy for an adult with cyanotic congenital heart disease. AB - Few patients with cyanotic congenital heart disease reach adulthood without a cardiac operation. The prognosis for ''unrepaired'' pulmonary atresia with ventricular septal defect is approximately 8% in the 1st decade of age. Consequently, the number of adults with this particular heart disease (unrepaired) who are expected to need a non-cardiac surgery is extremely low. General anesthesia may aggravate the preexisting right to left shunt and lead to persistent severe hypoxemia. The goal of anesthetic management should be to maintain intravascular volume. Systemic and pulmonary vascular resistance changes, such as might occur due to acidosis, hypothermia, hypercarbia or excessive airway pressures, should be avoided. Maintenance of preload, contractility and sinus rhythm is of major importance. The complex pathophysiology of such heart disease, in addition to the circumstances of emergency operation, exacerbate the total anesthetic risk. We present here a rare case of an acute appendectomy with successful outcome in an adult with pulmonary atresia and ventricular septal defect. PMID- 19190567 TI - Connecting active living research and public policy: transdisciplinary research and policy interventions to increase physical activity. AB - National and international organizations recommend creation of environments that support physical activity where people live, work, play, study, and travel. Policy changes can lead to activity-supportive environments and incentives. Research on environmental and policy influences on physical activity is well underway in many countries. An important use of the research is to inform policy debates, but the "translation" of research to policy is an emerging science. The papers in this supplement were presented at the 2008 Active Living Research Conference whose theme was "Connecting Active Living Research to Policy Solutions." The papers include evaluations of policy initiatives and research that suggests promising new policies. Commentaries propose principles for improving the translation of research to policy. Improving the rigor of research, asking policy-relevant questions, presenting country-specific data, and effectively communicating findings to policy makers are likely to contribute to greater impact of research on policy processes. PMID- 19190568 TI - Effect of innovative building design on physical activity. AB - Stair climbing can be a low-cost and relatively accessible way to add everyday physical activity, but many building stairwells are inaccessible or unpleasant and elevators are far more convenient. This study explores the use of and attitude toward stairs in an innovative office building where the main elevators for able-bodied users stop only at every third floor ("skip-stop" elevators). These users are expected to walk up or down nearby stairs that have been made open and appealing ("skip-stop" stairs). The study takes advantage of a natural experiment. Some workers' offices were clustered around the skip-stop elevator and the stairs, whereas others had access to a traditional elevator core, that is, an elevator that stopped at each floor with nearby fire exit stairs. Stair use on the open skip-stop stairs and enclosed fire stairs was measured using infrared monitors and card-reader activity logs. An online survey of employees (N=299, a 17.4% response rate) gathered information on stair use and attitudes and behaviors toward physical activity; interviews with key personnel identified major implementation issues. The skip-stop stair was used 33 times more than the enclosed stair of the traditional elevator core, with 72% of survey participants reporting daily stair use. Although implementation issues related to organizational objectives, costs, security, barrier-free accessibility, and building codes exist, the skip-stop feature offers a successful strategy for increasing stair use in workplaces. PMID- 19190569 TI - Arkansas Act 1220 of 2003 to reduce childhood obesity: its implementation and impact on child and adolescent body mass index. AB - Arkansas was among the first states to pass comprehensive legislation to combat childhood obesity, with Arkansas Act 1220 of 2003. Two distinct but complementary evaluations of the process, impact, and outcomes of Act 1220 are being conducted: first, surveillance of the weight status of Arkansas children and adolescents, using the statewide data amassed from the required measurements of students' body mass indexes (BMIs); and second, an independent evaluation of the process, impact, and outcomes associated with Act 1220. Various stakeholder groups initially expressed concerns about the Act, specifically concerns related to negative social and emotional consequences for students and an excessive demand on health care. Evaluation data, however, suggest that few adverse effects have occurred either in these areas of concern or in other concerns which have emerged over time. Schools are changing environments and implementing policies and programs to promote healthy behaviors and BMI levels have not increased since the implementation of Act 1220 in 2004. The Arkansas experience to date may serve to inform the efforts of other states to adopt policies to address the epidemic of childhood obesity. PMID- 19190570 TI - Early impact of the federally mandated Local Wellness Policy on physical activity in rural, low-income elementary schools in Colorado. AB - The What's Working project described the initial impact of the United States' federally mandated Local Wellness Policy in rural, low-income elementary schools located in Colorado. Before and after the Local Wellness Policy mandate went into effect, a survey about school features related to nutrition and physical activity was sent to a random sample of 45 rural elementary schools (i.e., schools located outside of urban areas), in which at least 40% of students qualified for free or reduced-cost lunch. Overall, opportunities for physical activity did not change after the policy went into effect: although time in physical education increased by 14 min per week (P=0.10), time for recess decreased by roughly 19 min per week (P=0.10). Policies supporting student participation in physical education and recess (an unstructured time during school hours when students are allowed to play outside) did not change. The researchers coded Local Wellness Policies and found them to have weak wording that produced minimal impact. Content analysis of key informant interviews suggested several barriers to the impact of the Local Wellness Policies: (1) competing pressures facing school districts, (2) lack of resources devoted to the Local Wellness Policy, (3) principals' lack of knowledge about the policy, and (4) lack of accountability mechanisms to ensure policy implementation. Financial resources and more effective communication about Local Wellness Policies among school districts and principals are needed to elevate the importance of and increase opportunities for physical activity in rural, low income Colorado elementary schools. PMID- 19190571 TI - Translating research into public policy. PMID- 19190572 TI - Preventing childhood obesity through state policy: qualitative assessment of enablers and barriers. AB - As the prevalence of obesity rapidly climbs among youth in the United States, public health practitioners and policymakers seek effective means of slowing and reversing these trends. Recently, many state laws and regulations addressing childhood obesity have been introduced and enacted. Understanding determinants of such legislation may inform the development and passage of future policies. For this study, key-informant interviews were conducted with 16 legislators and staffers from 11 states in 2005-2006 to examine qualitative factors that enable and impede state-level childhood obesity prevention legislation. Commonly cited factors positively influencing the passage of childhood obesity prevention legislation included national media exposure, introduction of the policy by senior legislators, and gaining the support of key players including parents, physicians, and schools. Noteworthy barriers included powerful lobbyists of companies that produce unhealthy foods and misconceptions about legislating foods at schools. Although the total number of informants was modest, their valuable insights provide policymakers and practitioners with a set of enablers and barriers to be considered when pursuing state-level policy. PMID- 19190573 TI - Correlates of walking to school and implications for public policies: survey results from parents of elementary school children in Austin, Texas. AB - Walking can be a healthy, sustainable, and equitable mode of transportation, but is not widely used for children's school travel. This study identifies multi level correlates of walking to/from school and relevant policy implications. We surveyed parents/guardians of 2,695 students from 19 elementary schools in Austin, Texas, which featured diverse sociodemographic and environmental characteristics. Among the personal and social factors, negative correlates were parents' education, car ownership, personal barriers, and school bus availability; positive correlates were parents' and children's positive attitude and regular walking behavior, and supportive peer influences. Of physical environmental factors, the strongest negative correlates were distance and safety concerns, followed by the presence of highways/freeways, convenience stores, office buildings, and bus stops en route. Our findings suggest that society should give high priority to lower socioeconomic status populations and to multi agency policy interventions that facilitate environmental changes, safety improvements, and educational programs targeting both parents and children. PMID- 19190574 TI - Sociodemographic, family, and environmental factors associated with active commuting to school among US adolescents. AB - Active commuting (non-motorized transport) to school can be an important source of physical activity for children and adolescents. This research examined sociodemographic, family, and environmental characteristics associated with active commuting to or from school among 3,451 US adolescents aged 12-17 years, who responded to the 2005 California Health Interview Survey. Logistic regression results indicated that those more likely to actively commute were males, Latinos, from lower-income families, attending public school, living in urban areas, and living closer to school. Adolescents without an adult present after school and those whose parents know little about their whereabouts after school were also more likely to actively commute. Parental walking for transportation and perceptions of neighborhood safety were not associated with adolescent active commuting. Important family and individual correlates of walking or biking to school among adolescents were identified, even after adjusting for distance to school and urbanicity. PMID- 19190575 TI - Can we achieve evidence-based policy and practice on active travel? PMID- 19190576 TI - Implementation of Texas Senate Bill 19 to increase physical activity in elementary schools. AB - In 2001, the 77th Texas legislature passed Senate Bill 19 requiring students in publicly funded elementary schools to participate in physical activity and schools to implement a coordinated school health program by September 1, 2007. We report on awareness of and adherence to Senate Bill 19 in a statewide sample of elementary schools and a subsample in two public health regions located along the Texas-Mexico border. Statewide, structured interviews with principals indicated high awareness of Senate Bill 19's requirements, but lower awareness of the need for parental involvement. Only 43% of Texas schools had adopted their coordinated program 1 year or less before the implementation deadline. Principals reported an average of 179 min of physical education per week, higher than the 135-min mandate. Among the subsample border schools, principals' physical activity reports were consistent with teacher records and student reports. Further, direct observation of physical education indicated that 50% of class time was spent in moderate and vigorous physical activity, meeting the level of physical activity intensity recommended by Healthy People 2010. The differences observed by public health regions included: more physical activity minutes in Region 10 (231 min compared to 217 min in Region 11); higher adoption of coordinated school health programs (92% compared to 75%); more district-level school health advisory committees (58% compared to 38%) and school-level school health advisory committees (83% compared to 25%); and a lower prevalence of obesity in 4th grade students (21% compared to 32%). Differences by region suggest that Senate Bill 19 is not being adhered to equally across the state, and some regions may require further support to increase implementation. Results underscore the importance of continued monitoring of enacted legislation, and that legislation for child health that focuses on school programs and policies requires funding and refinement to produce the intended effect. PMID- 19190578 TI - Social science, equal justice, and public health policy: lessons from Los Angeles. PMID- 19190577 TI - New recreational facilities for the young and the old in Los Angeles: policy and programming implications. AB - It is assumed that higher quality recreation facilities promote physical activity and serve communities better. We tested this assumption by comparing changes in the use of an expanded and renovated skate park (a facility for skateboarding) and a modernized senior citizen's center to two similar facilities that were not refurbished. The skate park was nearly tripled in size, and the senior center was remodeled and received new exercise equipment, a courtyard garden, and modern architectural features. We assessed use of these facilities through direct observation and surveyed both facility users and residents living within 2 miles of each facility. We found that making improvements to facilities alone will not always guarantee increased use. Although there was a 510% increase in use of the expanded skate park compared to a 77% increase in the comparison skate park, the senior center had substantially fewer users and provided fewer hours of exercise classes and other programmed activities after the facility was renovated. The implication of our study is that use results from a complex equation that includes not only higher quality recreation facilities but also progamming, staffing, fees, hours of operation, marketing, outreach, and perhaps a host of other human factors. PMID- 19190579 TI - Disparities in urban neighborhood conditions: evidence from GIS measures and field observation in New York City. AB - Although many low-income urban areas are highly walkable by conventional measures such as population density or land use mix, chronic diseases related to lack of physical activity are more common among residents of these areas. Disparities in neighborhood conditions may make poor areas less attractive environments for walking, offsetting the advantages of density and land use mix. This study compared poor and nonpoor neighborhoods in New York City, using geographic information systems measures constructed from public data for US census tracts within New York City (N=2,172) as well as field observation of a matched-pair sample of 76 block faces on commercial streets in poor and nonpoor neighborhoods. Poor census tracts had significantly fewer street trees, landmarked buildings, clean streets, and sidewalk cafes, and higher rates of felony complaints, narcotics arrests, and vehicular crashes. The field observation showed similar results. Improving aesthetic and safety conditions in poor neighborhoods may help reduce disparities in physical activity among urban residents. PMID- 19190580 TI - The relation of the perceived environment to fear, physical activity, and health in public housing developments: evidence from Chicago. AB - Within the realm of active living in urban neighborhoods in the United States, only a few studies have addressed the factors that promote or inhibit active living among residents in public housing. This paper examines the environmental and interpersonal factors associated with active living and health in public housing. We specifically examine the environmental predictors of fear of crime, and whether fear is related to moderate physical activity, and in turn, health and obesity. The analysis drew upon data from a sample of 328 African-American residents living in two severely distressed public housing developments in 2007. Structural equation modeling was used to calculate a path model of direct and indirect effects. Perceptions of violence were positively associated with fear, and both fear and physical activity predicted health. However, neither fear nor physical activity was associated with obesity. We discuss the implications of these findings for improving the quality of life of disadvantaged residents living in high-crime neighborhoods. PMID- 19190581 TI - Disparities in physical activity and sedentary behaviors among US children and adolescents: prevalence, correlates, and intervention implications. AB - Regular physical activity is important for health benefits among youth, but disparities exist. This paper describes disparities in physical activity participation and sedentary behaviors among youth in the United States, provides intervention implications, and offers recommendations for future research focused on reducing disparities related to levels of physical activity. Secondary analysis of national accelerometer data showed that achievement of recommended levels of physical activity ranged across subgroups from 2% to 61%. Mean hours per day spent in sedentary behavior ranged from 5.5 to 8.5. The largest disparities were by gender and age. An improved understanding of correlates may inform the design of interventions to increase physical activity in targeted subgroups. Additional theoretically based research is needed to elucidate which factors contributing to physical activity disparities are amenable to change via intervention. To eliminate health disparities, changes in policies that have an impact on physical activity may be necessary to promote physical activity among high-risk youth. PMID- 19190582 TI - Where different worlds collide: expanding the influence of research and researchers on policy. PMID- 19190583 TI - Factors associated with federal transportation funding for local pedestrian and bicycle programming and facilities. AB - Providing safe, convenient places for walking and bicycling can reduce barriers to participating in regular physical activity. We examined bicycle- and pedestrian-related investments authorized by federal transportation legislation in 3,140 counties in the United States by region, population size and urbanization, social and economic characteristics, and indicators of travel related walking and bicycling. From 1992 to 2004, states and counties implemented 10,012 bicycle- and pedestrian-related projects representing $3.17 billion in federal expenditures. We found disparities in implementation and system-building outcomes according to population size and location and social and economic indicators. Counties characterized by persistent poverty (odds ratio=0.69, 95% confidence interval 0.53-0.91) or low educational status (odds ratio=0.66, 95% confidence interval 0.52-0.84) were less likely to implement projects. Three key policy recommendations for improving public health outcomes are drawn from this research: Improved data tracking, more explicit linkages between transportation projects and public health, and improved planning assistance to underserved communities are all seen as essential steps. PMID- 19190584 TI - Transit and health: mode of transport, employer-sponsored public transit pass programs, and physical activity. AB - Increased provision of transit service and policy incentives that favor transit use can support a physically active lifestyle. We used the smartraq travel survey in metropolitan Atlanta, Georgia (in 2001-2002) to assess whether transit and car trips were associated with meeting the recommended levels of physical activity by using walking as a means of transportation. Additionally, we assessed associations between walking and using an employer-sponsored public transit pass. We controlled for demographics, neighborhood density, presence of services near workplaces, distance from home to transit, and car availability in our sample of 4,156 completed surveys. Walking distances from origin to destination were derived by a geographical information system and categorized as: no walking, moderate walking, or meeting recommendation (walking>or=2.4 km (1.5 miles) a day, approximately>or=30 min). In a multinomial logistic regression controlling for other covariates, transit trips were associated with an odds ratio (OR) of 3.87 (confidence interval (CI) 95%, 2.93-5.11) of meeting recommendation. In a multinominal logistical regression controlling for other covariates, transit users were associated with meeting recommendation, OR 2.23 (CI 95%, 1.27-3.90). PMID- 19190585 TI - Bicycling for transportation and health: the role of infrastructure. AB - This paper aims to provide insight on whether bicycling for everyday travel can help US adults meet the recommended levels of physical activity and what role public infrastructure may play in encouraging this activity. The study collected data on bicycling behavior from 166 regular cyclists in the Portland, Oregon metropolitan area using global positioning system (GPS) devices. Sixty percent of the cyclists rode for more than 150 minutes per week during the study and nearly all of the bicycling was for utilitarian purposes, not exercise. A disproportionate share of the bicycling occurred on streets with bicycle lanes, separate paths, or bicycle boulevards. The data support the need for well connected neighborhood streets and a network of bicycle-specific infrastructure to encourage more bicycling among adults. This can be accomplished through comprehensive planning, regulation, and funding. PMID- 19190586 TI - Emergency surgical treatment of advanced endomyocardial fibrosis in Mozambique. AB - BACKGROUND: An 11-year-old girl presented to a specialist cardiac facility in Mozambique. She had severe heart failure and massive cardiac enlargement, herniation of the heart into the epigastrium, atrial fibrillation, signs of severe pulmonary hypertension and a low cardiac output. INVESTIGATIONS: Chest radiography, echocardiography, 24 h Holter monitoring, and cardiac catheterization. DIAGNOSIS: Left and right endomyocardial fibrosis in conjunction with an aneurysmal left atrium, severe heart failure, and atrial fibrillation. MANAGEMENT: Left ventricular endocardiectomy with mobilization of the chordae tendineae, mitral valve repair, tricuspid annuloplasty, and left atrial resection. PMID- 19190587 TI - Do tumor characteristics predict risk of malignancy in thyroid nodules with indeterminate cytology? AB - The use of ultrasonography-guided, fine-needle aspiration cytology (FNAC) has enhanced the preoperative characterization of thyroid nodules and reduced the need for diagnostic surgeries. However, complete preoperative characterization is not yet possible for patients with an indeterminate FNAC result. Here, results are discussed from a retrospective study by Banks et al. that aimed to refine preoperative risk assessment of thyroid cancer in patients with indeterminate FNAC results. The authors found that patients' age, nodule size and specific cytological diagnoses were all associated with thyroid malignancy. These variables were used to construct a diagnostic predictor model. The study by Banks et al. provides general guidance for clinicians who need to estimate thyroid cancer risk for patients with indeterminate FNAC results. Refinements in risk assessment for thyroid malignancy might help to individualize care of these patients; however, the results also underscore the need for enhanced diagnostic methods. PMID- 19190588 TI - Does intensive glycemic control for type 2 diabetes mellitus have long-term benefits for cardiovascular disease risk? AB - This Practice Point commentary discusses the findings of the 10-year follow-up of the United Kingdom Prospective Diabetes Study, which was a multicenter, randomized, controlled trial in which patients with newly diagnosed type 2 diabetes mellitus received conventional or intensive glycemic control. The follow up study by Holman et al. suggests that intensive glycemic control can have long lasting benefits in reducing the incidence not only of diabetes-related end points and microvascular complications (as shown in the original UKPDS study) but also of myocardial infarction and death from any cause. These benefits occurred despite the early loss of within-trial differences in HbA(1c) levels between the intensive and conventional treatment groups. The findings of Holman et al. support the notion of a sustained, legacy effect of intensive glycemic control, which was originally suggested in the Epidemiology of Diabetes Interventions and Complications study. PMID- 19190589 TI - Predicting response to acetylcholinesterase inhibitor treatment in Alzheimer disease: has the time come? AB - Identification of both the initial and the sustained clinical benefits achieved with symptomatic treatments for Alzheimer disease (AD) is a challenge. This commentary addresses a report by Wattmo et al. on 3-year follow-up of a cohort of patients with AD who were treated with donepezil. The investigators developed predictive regression models that can accurately calculate group mean Alzheimer's Disease Assessment Scale-cognitive subscale (ADAS-cog) and Mini-Mental State Examination (MMSE) scores. They determined that patients with mild to moderate AD have a mean 5-7-month cognitive improvement with donepezil treatment, with greater benefit in more-advanced disease. While these results are encouraging, this study has important limitations. Although the predictive models work well for determining group means, the authors note that they do not predict individual patient responses, which vary greatly. Additionally, the study had a drop-out rate of 62%, which might elicit survivorship bias and overestimation of treatment benefit. We remind clinicians that small improvements in cognition matter most when a concurrent measurable benefit is seen in daily function. PMID- 19190590 TI - Cetuximab therapy for patients with advanced squamous cell carcinomas of the head and neck. AB - We discuss the results of the phase III EXTREME (Erbitux in First-Line Treatment of Recurrent or Metastatic Head and Neck Cancer) trial. In this study, 442 untreated patients with advanced squamous cell carcinomas of the head and neck (SCCHN) were randomly assigned to receive platinum and 5-fluorouracil with or without cetuximab. Median overall survival -- the primary end point of the trial - was improved in the cetuximab arm compared with the chemotherapy-alone arm (10.1 months and 7.4 months, respectively; P = 0.04). This is the first phase III trial in over two decades to exhibit a survival advantage in patients with SCCHN not amenable for curative treatment; however, it raises several considerations for clinical practice and future research directions, such as the optimum choice of chemotherapy to be combined with cetuximab, sequencing of cetuximab with chemotherapy, predictive markers of benefit from cetuximab, as well as implications for patients with locally advanced, potentially curable disease. PMID- 19190591 TI - Treatment of advanced disease in patients with well-differentiated neuroendocrine tumors. AB - Well-differentiated neuroendocrine tumors (NETs), which are also referred to as well-differentiated endocrine carcinoma according to WHO terminology, are usually slow-growing cancers, even when they exhibit gross local invasion and/or metastases. The survival of patients with metastatic NET is often measured in years to decades. Once NET progresses or becomes symptomatic the patient's prognosis is poor. An important challenge for clinicians is to distinguish at an early stage those patients who will die with the disease, from those who will succumb because of it, so that the appropriate level of care can be administered. Reliable genomic predictors could provide substantial advancements in prognosis and, possibly, treatment; however, such markers are currently unavailable. Early literature on the treatment of NETs is confounded by a lack of formal objective response criteria. Somatostatin analogs can control symptoms and can stabilize some slow-growing tumors, but rarely result in tumor regression. Surgery is curative in only a minority of patients, and systemic chemotherapy is minimally effective. Advances in the understanding of tumor biology have led to the identification of important cellular processes involved in the pathogenesis of NETs, and agents that target these processes have now entered clinical trials. We will discuss the data on therapies currently used to treat well-differentiated NETs, and the strategies being used in clinical trials. PMID- 19190593 TI - Health-related quality-of-life effects of watchful waiting re-evaluated in SPCG 4. AB - This commentary discusses the re-analysis of the Scandinavian Prostate Cancer Group Study Number 4 (SPCG-4), which had previously shown a modest but significant survival benefit of prostatectomy over watchful waiting. In the re analysis, patients allocated to watchful waiting experienced a significantly decreased health-related quality of life (HRQOL) at 6-8 years after randomization, compared with those who underwent radical prostatectomy. HRQOL differences between treatment arms in the first 5 years after randomization were limited to worse erectile dysfunction among patients randomized to prostatectomy. These findings indicate that watchful waiting might be associated with greater late deterioration in select domains of HRQOL than is prostatectomy. In addition, patients randomized to watchful waiting who received androgen deprivation therapy (ADT) reported a lower sense of well-being, more-depressed mood, and a lower self assessed quality of life compared with those who did not receive ADT, which indicates that ADT might adversely affect HRQOL in this patient population. PMID- 19190592 TI - Effect of bisphosphonates on pain and quality of life in patients with bone metastases. AB - Bone is the most common organ for tumor metastasis, especially in patients with cancers of the breast or prostate. Bone metastases disrupt skeletal metabolism and result in considerable skeletal morbidity, including intractable, chronic bone pain, hypercalcemia of malignancy, pathologic fracture and spinal-cord compression. In addition to the chronic pain caused by bone metastases, skeletal related events (SREs) such as pathologic fractures and spinal-cord compression can result in acute increases in pain. These effects can severely impair mobility and contribute to a general decrease in quality of life. Palliative options to treat bone metastases include radiotherapy, analgesics, surgery and bisphosphonates. These drugs bind to the surface of the bone and impair osteoclast-mediated bone resorption, and reduce the tumor-associated osteolysis that is initiated by the development of skeletal metastases. In addition to preventing SREs, bisphosphonates can palliate bone pain caused by a variety of solid tumors. This Review summarizes the clinical trial data of bisphosphonates for the prevention of SREs and the palliation of bone pain. Among these agents, nitrogen-containing bisphosphonates are recognized as the most effective, and zoledronic acid has demonstrated the broadest clinical utility. PMID- 19190594 TI - Does pelvic floor muscle training prevent and treat urinary and fecal incontinence in pregnancy? AB - This Practice Point commentary discusses the systematic review by Hay-Smith and colleagues, which investigated the efficacy of pelvic floor muscle training (PFMT) for preventing and treating urinary and fecal incontinence in pregnant and postnatal women. The authors concluded that antenatal PFMT improved continence outcomes during first pregnancy, and that PFMT represents an appropriate treatment for women with persistent postpartum incontinence. This systematic review is of high methodological quality. Nevertheless, the exclusion of studies with nonrandomized designs and the inclusion of large studies of weak interventions might have resulted in an underestimation of the effect of PFMT. The dose-response relationship is important in PFMT, and intensive training with close follow-up is required to achieve beneficial effects. PMID- 19190596 TI - Crohn's colitis versus ulcerative colitis: should surveillance for dysplasia and cancer differ? AB - Although the risk of developing dysplasia and carcinomas in the colon is known to increase over time in patients with ulcerative colitis, data are scarce on the need for surveillance colonoscopies in patients with long-standing Crohn's colitis. This article discusses the results of a study by Friedman et al. conducted in 259 patients with chronic Crohn's colitis who received a median of four follow-up colonoscopies over 25 years after an initial negative examination result for dysplasia or carcinomas. The cumulative risk for developing dysplasia or carcinomas has only increased from 22% to 25% in the 10 years since the previous report from this study's authors, and the prevalence of extensive colitis was higher in study participants than that expected in a random population of patients with Crohn's colitis. However, Friedman et al. provide convincing evidence that long-term surveillance for the development of colonic dysplasia and cancers is advantageous in this population. PMID- 19190597 TI - Behavioral therapy for IBS. PMID- 19190595 TI - Inhibition of multidrug-resistant Acinetobacter baumannii by nonviral expression of hCAP-18 in a bioengineered human skin tissue. AB - When skin is compromised, a cascade of signals initiates the rapid repair of the epidermis to prevent fluid loss and provide defense against invading microbes. During this response, keratinocytes produce host defense peptides (HDPs) that have antimicrobial activity against a diverse set of pathogens. Using nonviral vectors we have genetically modified the novel, nontumorigenic, pathogen-free human keratinocyte progenitor cell line (NIKS) to express the human cathelicidin HDP in a tissue-specific manner. NIKS skin tissue that expresses elevated levels of cathelicidin possesses key histological features of normal epidermis and displays enhanced antimicrobial activity against bacteria in vitro. Moreover, in an in vivo infected burn wound model, this tissue results in a two log reduction in a clinical isolate of multidrug-resistant Acinetobacter baumannii. Taken together, these results suggest that this genetically engineered human tissue could be applied to burns and ulcers to counteract bacterial contamination and prevent infection. PMID- 19190598 TI - A case of Cowden's syndrome presenting with gastric carcinomas and gastrointestinal polyposis. AB - BACKGROUND: A 73-year-old white man was referred to a cancer genetics clinic for evaluation of a approximately 20-year history of mixed upper and lower gastrointestinal polyposis, including hyperplastic, inflammatory and adenomatous polyps, colonic ganglioneuromas, and associated diffuse, esophageal glycogenic acanthosis. Two synchronous gastric carcinomas had been identified before referral and the patient had undergone a total gastrectomy, omentectomy and cholecystectomy. Multiple hyperplastic polyps and small, sessile polyps were also observed in the gastrectomy specimen. INVESTIGATIONS: History and physical examination, upper and lower gastrointestinal endoscopy and biopsy, genetic testing, molecular pathology investigations (immunohistochemistry), thyroid ultrasonography, fine-needle aspiration of a thyroid nodule. DIAGNOSIS: Cowden's syndrome. MANAGEMENT: Genetic counseling, thyroidectomy, vitamin B(12) supplementation, continued endoscopic surveillance and genetic testing of at-risk family members. PMID- 19190603 TI - Pro: a large colonic polyp is best removed by laparoscopy. PMID- 19190600 TI - Pathogenesis of portal hypertensive gastropathy: translating basic research into clinical practice. AB - Portal hypertensive gastropathy (PHG) is often seen in patients with portal hypertension, and can lead to transfusion-dependent anemia as well as acute, life threatening bleeding episodes. This Review focuses on the mechanisms that underlie the pathogenesis of PHG that provide reasonable grounds for the treatment of this condition, and ultimately enable translation of basic research into clinical practice. Increased portal pressure associated with cirrhosis and liver dysfunction is critical for the development of clinically significant PHG, and leads to impaired gastric mucosal defense mechanisms that render the stomach susceptible to mucosal injury. The use of pharmacological agents such as beta blockers reduces the frequency of bleeding episodes in PHG. As a last resort, surgical decompression of the portal system, transjugular intrahepatic stent placement and liver transplantation can resolve this condition. Elimination of known risk factors for gastric injury such as alcohol, aspirin and traditional NSAIDs is critical. The role of Helicobacter pylori colonization of the gastric mucosa in PHG is not clear. Careful and critical interpretation of human and experimental data can be helpful to establish a rationale for the medical management of this important condition. PMID- 19190599 TI - Multimodal approaches to the treatment of hepatocellular carcinoma. AB - The prevalence of hepatocellular carcinoma in Europe and the US is increasing and is currently the leading cause of death in patients with cirrhosis. Surveillance programs for patients with cirrhosis aim to detect tumors at an early stage, when the greatest therapeutic benefits can be achieved. Curative treatments for early stage tumors include liver transplantation, resection and percutaneous ablation. Transarterial chemoembolization (TACE) and sorafenib can improve survival for patients with intermediate and advanced tumors, respectively. In clinical practice, combination therapies are often used, despite limited evidence to support this approach from randomized controlled trials. Combination therapy with radiofrequency ablation (RFA) plus percutaneous ethanol injection can, however, improve survival for selected patients compared with RFA alone. Combined treatment with TACE and RFA also improves patients' survival compared with TACE or RFA monotherapy. TACE performed before or after surgical resection, however, is not beneficial. Prevention of tumor progression in patients awaiting liver transplantation requires nonsurgical treatments; however, the real advantages of the available treatment modalities are yet to be defined. The role of sorafenib administration in combination with TACE after the use of potentially curative treatments, for the treatment of intermediate hepatocellular carcinoma, or in selective settings after liver transplantation, requires further study. PMID- 19190604 TI - Con: colonoscopic resection of large neoplastic lesions is appropriate and safe. PMID- 19190605 TI - A balancing view: should large colon polyps be removed colonoscopically or surgically? PMID- 19190606 TI - Non-acid reflux: detection by multichannel intraluminal impedance and pH, clinical significance and management. PMID- 19190607 TI - Globus: are we any nearer to understanding it? AB - Globus is a common functional symptom without a clearly accepted etiology. Upper esophageal sphincter (UES) hypertension and gastroesophageal reflux have been proposed but not confirmed. Kwiatek et al. report a detailed study in globus patients using high-resolution manometry (HRM). The study showed greater respiratory augmentation of UES pressure compared with control groups among two thirds of globus patients. Further prospective studies are needed to confirm these findings and to establish whether they are of etiologic significance. PMID- 19190608 TI - Auditory evoked potentials compared with bispectral index for monitoring of midazolam and propofol sedation during colonoscopy. AB - OBJECTIVES: The purpose of this study was to evaluate and compare Bispectral index (BIS) and A-line auditory evoked potential index (AAI) for monitoring depth of low-dose midazolam and propofol sedation during colonoscopy. METHODS: A total of 115 consecutive patients (ASA I-IV), receiving low-dose midazolam and propofol sedation for colonoscopy, were evaluated. BIS and AAI levels, Observer's Assessment of Alertness/Sedation (OAA/S) scores, blood pressure, heart rate, oxygen saturation, as well as the presence or absence of eyelash reflex, patient reaction to an external noxious stimulus and to procedure-related pain were recorded every 1-3 min by a single trained observer. RESULTS: There was a positive correlation between BIS and OAA/S scores (correlation coefficient=0.77) and to a lesser extent AAI and OAA/S scores (correlation coefficient=0.47). BIS and AAI showed significant differences between subsequent levels of sedation (P<0.001). The clustered receiver operating characteristic curve estimate of BIS for the detection of deep sedation was significantly better than that of AAI (P<0.001). Regarding the presence or absence of eyelash reflex and patient reaction to an external noxious stimulus and to procedure-related pain, significant different levels were found for BIS as well as AAI, respectively. Only small changes were observed in hemodynamic variables and oxygen saturation. Overall, our data suggest target BIS levels of slightly above 73 for moderate sedation (defined as OAA/S scores 2 and 3). CONCLUSIONS: BIS and AAI correlated with the level of sedation. Hemodynamic variables were poor indicators of the hypnotic-anesthetic status of the patient. BIS discriminated best between moderate and deep sedation and could complement clinical observation for guidance of moderate sedation. PMID- 19190609 TI - Management of painful chronic pancreatitis with single-dose radiotherapy. AB - OBJECTIVES: Some patients with chronic pancreatitis present recurrent flare-ups of pancreatitis and/or unrelenting pain. Current management is mostly limited to analgesics and surgery. We reasoned that anti-inflammatory radiotherapy, which has proven useful to alleviate other painful inflammatory painful disorders, might prove valuable for severely symptomatic patients with chronic pancreatitis. METHODS: We prospectively studied the efficacy of single-dose anti-inflammatory radiotherapy in 15 consecutive patients with chronic pancreatitis who fulfilled the following criteria: either two flare-ups of pancreatitis in the previous 6 months and/or continuous pain for more than 3 months. Treatment consisted of a single radiation dose of 8 Gy to the pancreas. Exocrine function (fecal elastase), endocrine function (c peptide), quality of life (EuroQol questionnaire), and clinical outcome were assessed before and after radiation. Response was defined as no further pain or flare-ups of pancreatitis. RESULTS: During follow-up (median: 39 months; range: 4-72 months), 12 patients had no further pain or flare-ups. One patient required a second radiation dose 1 year after the initial treatment, but he has remained well ever since (50 months). Two other patients did not respond to radiotherapy. After radiotherapy either exocrine or endocrine pancreatic function, or both, deteriorated in three patients. Patients who responded to treatment (13/15) gained 4-20 kg in body weight during follow-up (median 4 kg) and EuroQol improved significantly from 0.58 to 0.86 (P<0.001). CONCLUSIONS: Radiotherapy for severe symptomatic chronic pancreatitis appears to be a useful and effective therapeutic choice that could potentially substitute for or delay surgery. PMID- 19190610 TI - The utility of an evidence-based lecture and clinical prompt as methods to improve quality of care in colorectal cancer screening. AB - OBJECTIVES: Colorectal cancer screening can decrease both the incidence of and mortality from colorectal cancer. Unfortunately, participation rates remain suboptimal. We assessed whether an evidence-based lecture or a written clinical prompt at the time of a patient visit would independently increase colorectal cancer screening rates in an internal medicine resident clinic. METHODS: Three phase prospective cohort trial of 750 patients. The first phase assessed the baseline screening rate. The second phase assessed the screening rate after an evidenced-based lecture. The third phase assessed the screening rate after the addition of written prompts to patient charts. All 50- to 80-year-old patients who met the criteria for colorectal cancer screening were included in the study. The first intervention was a 1-h evidence-based lecture addressing colorectal cancer screening. The second intervention was placing a written prompt on all eligible patients' charts, reminding physicians to assess whether their patient had been screened. Demographic characteristics were assessed for each cohort of 250 patients. The percentage of patients with documented intention to be screened was assessed for each cohort. RESULTS: The evidence-based lecture did not significantly improve overall attempted screening rates or colon imaging rates relative to baseline. The clinical prompt significantly improved attempts at screening relative to baseline (39.6 to 67.6%) (P<0.0001). Ordering of colon imaging rates also significantly improved after instituting the prompt from 24 to 46% (P<0.0001). CONCLUSIONS: Clinical prompts are superior to evidence-based lectures for improving physician colorectal cancer screening practices. These prompts are simple low-cost measures that can improve quality of care. PMID- 19190616 TI - Spontaneous rupture of gastroduodenal artery pseudoaneurysm following vigorous cough. PMID- 19190617 TI - Consecutively acquired sexually transmitted infections mimicking Crohn's disease. PMID- 19190621 TI - Are static resting wrist splints beneficial in early RA? AB - In a randomized controlled trial, Adams et al. compared the effectiveness of static resting wrist splints plus occupational therapy with occupational therapy alone in 120 patients with early rheumatoid arthritis. Self-reported adherence with splint wear was moderate, with 24.5% of patients reporting that they had never worn the splints over the 12-month study period. Except for a small benefit of resting splints in the occurrence, but not the duration, of hand morning stiffness, there were no significant differences in structural impairment and functional hand ability outcomes between the control and splint groups. The findings of this trial are in line with the negative outcomes of earlier studies of static resting wrist splints in patients with longer disease duration. The currently available data suggest, therefore, that resting wrist splints might not be an effective routine treatment for patients with rheumatoid arthritis. PMID- 19190622 TI - A new 5-lipoxygenase inhibitor seems to be safe and effective for the treatment of osteoarthritis. AB - There is a clear need for both disease-modifying agents and alternative analgesic therapies in osteoarthritis. Currently, there are none of the former, and the latter are limited by adverse effects, particularly gastrointestinal and cardiovascular effects. A new class of analgesic agent that is under investigation inhibits 5-lipoxygenase (5-LOX), the enzyme that catalyzes the conversion of membrane-bound arachidonic acid to leukotrienes. As leukotrienes are implicated in a wide variety of pathologies, the potential of 5-LOX inhibitors has been explored in conditions as diverse as asthma, acute mountain sickness and coronary artery disease. A new 5-LOX inhibitor, derived from a herb, has undergone a phase II trial in osteoarthritis with promising results. Although a putative mechanism of action suggests a disease-modifying effect, the important outcomes from this trial are good symptom response and a low adverse effect profile, albeit in the small number of patients studied. PMID- 19190620 TI - Central and peripheral regulation of food intake and physical activity: pathways and genes. AB - A changing environment and lifestyle on the background of evolutionary engraved and perinatally imprinted physiological response patterns is the foremost explanation for the current obesity epidemic. However, it is not clear what the mechanisms are by which the modern environment overrides the physiological controls of appetite and homeostatic body-weight regulation. Food intake and energy expenditure are controlled by complex, redundant, and distributed neural systems involving thousands of genes and reflecting the fundamental biological importance of adequate nutrient supply and energy balance. There has been much progress in identifying the important role of hypothalamus and caudal brainstem in the various hormonal and neural mechanisms by which the brain informs itself about availability of ingested and stored nutrients and, in turn, generates behavioral, autonomic, and endocrine output. Some of the genes involved in this "homeostatic" regulator are crucial for energy balance as manifested in the well known monogenic obesity models. However, it can be clearly demonstrated that much larger portions of the nervous system of animals and humans, including the cortex, basal ganglia, and the limbic system, are concerned with the procurement of food as a basic and evolutionarily conserved survival mechanism to defend the lower limits of adiposity. By forming representations and reward expectancies through processes of learning and memory, these systems evolved to engage powerful emotions for guaranteed supply with, and ingestion of, beneficial foods from a sparse and often hostile environment. They are now simply overwhelmed with an abundance of food and food cues no longer contested by predators and interrupted by famines. The anatomy, chemistry, and functions of these elaborate neural systems and their interactions with the "homeostatic" regulator in the hypothalamus are poorly understood, and many of the genes involved are either unknown or not well characterized. This is regrettable because these systems are directly and primarily involved in the interactions of the modern environment and lifestyle with the human body. They are no less "physiological" than metabolic regulatory mechanisms that have attracted most of the research during the past 15 years. PMID- 19190623 TI - Certolizumab pegol--what role does this new TNF inhibitor have in the treatment of RA? AB - The efficacy and safety of a new tumor necrosis factor inhibitor, certolizumab pegol, in active rheumatoid arthritis has now been assessed in three phase III, multicenter, randomized, double-blind, placebo-controlled clinical trials. This commentary focuses on the paper by Keystone et al., in which patients were followed for the longest duration. This study, which compared two doses of subcutaneous certolizumab pegol with placebo in patients with active RA receiving methotrexate, showed no advantage of 400 mg over 200 mg certolizumab pegol over 52 weeks, after induction with 400 mg. The nature of the patients enrolled in this study, trial design and possible safety issues are discussed, as is whether this trial can teach us anything about tumor necrosis factor inhibitors in general. On the basis of the results from this study, certolizumab pegol does not represent a major addition to our armamentarium, but because of the slightly different mechanism of action and structure of this drug, and the apparently acceptable therapeutic effects over one year, it is, nevertheless, welcome. PMID- 19190624 TI - Clinical and radiographic response to a local infliximab injection in a patient with chronic sacroiliitis. AB - BACKGROUND: A 49-year-old man presented to a rheumatology clinic with a 2-month history of low back pain. The pain seemed to be inflammatory in origin, with nocturnal occurrence and substantial early morning stiffness. The patient was previously well, with no history of psoriasis, inflammatory bowel disease or iritis. INVESTIGATIONS: Physical examination, anteroposterior X-ray of the pelvis, isotope bone scan, CT and MRI of the sacroiliac joints, measurement of serum inflammatory markers, HLA-B27 testing, routine and mycobacterial culture of the sacroiliac joint fluid aspirate, tuberculosis skin testing and chest X-ray. DIAGNOSIS: An HLA-B27-negative patient with chronic unilateral sacroiliitis refractory to conventional management. MANAGEMENT: After unsuccessful treatment with various NSAIDs and physical therapy, local corticosteroid injections on two occasions in the affected sacroiliac joint provided transient relief. A subsequent 20 mg injection of infliximab into the affected joint resulted in clinical and radiological improvement that has been sustained for more than 2 years. PMID- 19190625 TI - Skin problems in chronic kidney disease. AB - Skin disorders associated with chronic kidney disease (CKD) can markedly affect a patient's quality of life and can negatively impact their mental and physical health. Uremic pruritus, which is frequently encountered in patients with CKD, is considered to be an inflammatory systemic disease rather than a local skin disorder. Biomarkers of inflammation are increased in patients with uremic pruritus and an imbalance of the endogenous opioidergic system might be involved in the complex pathogenesis of the disease. Treatment options for uremic pruritus include emollients, topical capsaicin cream, ultraviolet B phototherapy, gabapentin, oral activated charcoal and nalfurafine, a kappa-opioid-receptor agonist. Calcific uremic arteriolopathy is triggered by an imbalance of promoters and inhibitors of vascular calcification, caused by the inflammatory changes that occur in uremia. Promising therapeutic strategies for calcific uremic arteriolopathy include bisphosphonates and intravenous sodium thiosulfate. Nephrogenic systemic fibrosis is a devastating condition associated with the use of gadolinium-based contrast agents in patients with CKD. At present, no therapies are available for this complication. Preventive measures include use of iodine-based contrast agents, particularly in patients with CKD stage 4 and 5. If gadolinium contrast is necessary, administration of low volumes of the more stable macrocyclic ionic types of gadolinium-based contrast agent is advocated. Hemodialysis following gadolinium exposure might offer benefits but evidence is lacking. PMID- 19190626 TI - Regulation of focal adhesion turnover by ErbB signalling in invasive breast cancer cells. AB - A crucial early event by which cancer cells switch from localised to invasive phenotype is initiated by the acquisition of autonomous motile properties; a process driven by dynamic assembly and disassembly of multiple focal adhesion (FA) proteins, which mediate cell-matrix attachments, extracellular matrix degradation, and serve as traction sites for cell motility. We have reported previously that cancer cell invasion induced by overexpression of members of the ErbB tyrosine kinase receptors, including ErbB2, is dependent on FA signalling through FA kinase (FAK). Here, we show that ErbB2 receptor signalling regulates FA turnover, and cell migration and invasion through the Src-FAK pathway. Inhibition of the Src-FAK signalling in ErbB2-positive cells by Herceptin or RNA interference selectively regulates FA turnover, leading to enhanced number and size of peripherally localised adhesions and inhibition of cell invasion. Inhibition of ErbB2 signalling failed to regulate FA and cell migration and invasion in cells lacking FAK or Src but gains this activity after restoration of these proteins. Taken together, our results show a regulation of FA turnover by ErbB2 signalling. PMID- 19190627 TI - Cumulative survival in early-onset unilateral and bilateral breast cancer: an analysis of 1907 Taiwanese women. AB - As the epidemiological pattern of breast cancer in modernising Asian countries differs greatly from that in Western countries, it is worthwhile to investigate the long-term prognoses of unilateral and bilateral breast cancer in these nations. A retrospective cohort study composed of 1907 Taiwanese women was conducted to follow 1863 unilateral and 44 bilateral cases of breast cancer. Time dependent Cox regression was used to assess the risk of breast cancer death by considering the time course of unilateral and bilateral tumour development. The 15-year survival rates were 68.37, 62.63, and 26.42% for unilateral, synchronous bilateral, and metachronous bilateral breast cancer, respectively. Differences among types were most apparent after 5 years of follow-up. After adjusting for significant prognostic factors, the risk of death for overall bilateral breast cancer was 2.50-fold greater (95% CI, 1.43-4.37) compared to unilateral breast cancer. The corresponding figures were 1.12-fold (95% CI, 0.42-3.02) and 6.11 fold (95% CI, 3.14-11.89) for synchronous and metachronous bilateral breast cancer, respectively. Taiwanese women, who are frequently diagnosed with breast cancer before 50 years of age, showed poorer survival for metachronous bilateral than for synchronous bilateral or unilateral breast cancer. Survival was markedly poorer compared to recent data from Sweden. PMID- 19190628 TI - Breast cancer risk in elderly women with systemic autoimmune rheumatic diseases: a population-based case-control study. AB - Systemic autoimmune rheumatic diseases (SARDs) are chronic inflammatory and immuno-modulatory conditions that have been suggested to affect cancer risk. Using the Surveillance, Epidemiology and End Results-Medicare-linked database, women aged 67-99 years and diagnosed with incident breast cancer in 1993-2002 (n=84 778) were compared with an equal number of age-matched cancer-free female controls. Diagnoses of SARDs, including rheumatoid arthritis (RA, n=5238), systemic lupus erythematosus (SLE, n=340), Sjogren's syndrome (n=374), systemic sclerosis (n=128), and dermatomyositis (n=31), were determined from claim files for individuals from age 65 years to 1 year before selection. Associations of SARD diagnoses with breast cancer, overall and by oestrogen receptor (ER) expression, were assessed using odds ratio (OR) estimates from multivariable logistic regression models. The women diagnosed with RA were less likely to develop breast cancer (OR=0.87, 95% confidence interval (CI)=0.82-0.93). The risk reduction did not differ by tumour ER-status (OR=0.83, 95% CI=0.78-0.89 for ER positive vs OR=0.91, 95% CI=0.81-1.04 for ER-negative, P for heterogeneity=0.14). The breast cancer risk was not associated with any of the other SARDs, except for a risk reduction of ER-negative cases (OR=0.49, 95% CI=0.26-0.93) among women with SLE. These findings suggest that systemic inflammation may affect breast epithelial neoplasia. PMID- 19190629 TI - Carbogen breathing increases prostate cancer oxygenation: a translational MRI study in murine xenografts and humans. AB - Hypoxia has been associated with poor local tumour control and relapse in many cancer sites, including carcinoma of the prostate. This translational study tests whether breathing carbogen gas improves the oxygenation of human prostate carcinoma xenografts in mice and in human patients with prostate cancer. A total of 23 DU145 tumour-bearing mice, 17 PC3 tumour-bearing mice and 17 human patients with prostate cancer were investigated. Intrinsic susceptibility-weighted MRI was performed before and during a period of carbogen gas breathing. Quantitative R(2)* pixel maps were produced for each tumour and at each time point and changes in R(2)* induced by carbogen were determined. There was a mean reduction in R(2)* of 6.4% (P=0.003) for DU145 xenografts and 5.8% (P=0.007) for PC3 xenografts. In all, 14 human subjects were evaluable; 64% had reductions in tumour R(2)* during carbogen inhalation with a mean reduction of 21.6% (P=0.0005). Decreases in prostate tumour R(2)* in both animal models and human patients as a result of carbogen inhalation suggests the presence of significant hypoxia. The finding that carbogen gas breathing improves prostate tumour oxygenation provides a rationale for testing the radiosensitising effects of combining carbogen gas breathing with radiotherapy in prostate cancer patients. PMID- 19190630 TI - Does type 2 diabetes influence the risk of oesophageal adenocarcinoma? AB - Since hyperinsulinaemia may promote obesity-linked cancers, we compared type 2 diabetes prevalence among oesophageal adenocarcinoma (OAC) patients and population controls. Diabetes increased the risk of OAC (adjusted odds ratio 1.59, 95% confidence interval (CI) 1.04-2.43), although the risk was attenuated after further adjusting for body mass index (1.32, 95% CI 0.85-2.05). PMID- 19190631 TI - Overexpressed vs mutated Kras in murine fibroblasts: a molecular phenotyping study. AB - Ras acts in signalling pathways regulating the activity of multiple cellular functions including cell proliferation, differentiation, and apoptosis. Amino acid exchanges at position 12, 13, or 61 of the Kras gene convert the proto oncogene into an activated oncogene. Until now, a direct comparison of genome wide expression profiling studies of Kras overexpression and different Kras mutant forms in a single assay system has not been carried out. In our study, we focused on the direct comparison of global gene expression effects caused by mutations in codon 12 or 13 of the Kras gene and Kras overexpression in murine fibroblasts. We determined Kras cellular mRNA, Ras protein and activated Ras protein levels. Further, we compared our data to the proteome analysis of the same transfected cell lines. Both overexpression and mutations of Kras lead to common altered gene expression patterns. Only two genes, Lox and Col1a1, were reversely regulated in the Kras transfectants. They may contribute to the higher aggressiveness of the Kras codon 12 mutation in tumour progression. The functional annotation of differentially expressed genes revealed a high frequency of proteins involved in tumour growth and angiogenesis. These data further support the important role of these genes in tumour-associated angiogenesis. PMID- 19190632 TI - Gemcitabine-oxaliplatin combination for ovarian cancer resistant to taxane platinum treatment: a phase II study from the GINECO group. AB - Advanced ovarian carcinoma in early progression (<6 months) (AOCEP) is considered resistant to most cytotoxic drugs. Gemcitabine (GE) and oxaliplatin (OXA) have shown single-agent activity in relapsed ovarian cancer. Their combination was tested in patients with AOCEP in phase II study. Fifty patients pre-treated with platinum-taxane received q3w administration of OXA (100 mg m(-2), d1) and GE (1000 mg m(-2), d1, d8, 100-min infusion). Patient characteristics were a : median age 64 years (range 46-79),and 1 (84%) or 2 (16%) earlier lines of treatment. Haematological toxicity included grade 3-4 neutropaenia (33%), anaemia (8%), and thrombocytopaenia (19%). Febrile neutropaenia occurred in 3%. Non haematological toxicity included grade 2-3 nausea or vomiting (34%), grade 3 fatigue (25%),and grade 2 alopecia (24%). Eighteen (37%) patients experienced response. Median progression-free (PF) and overall survivals (OS) were 4.6 and 11.4 months, respectively. The OXA-GE combination has high activity and acceptable toxicity in AOCEP patients. A comparison of the doublet OXA-GE with single-agent treatment is warranted. PMID- 19190633 TI - The risk of febrile neutropenia in patients with non-small-cell lung cancer treated with docetaxel: a systematic review and meta-analysis. AB - We aimed to assess the incidence of febrile neutropenia in patients with non small cell lung cancer treated with docetaxel as second line chemotherapy by systematic review and meta-analysis of clinical studies. Published studies were retrieved and included if they considered docetaxel at the licensed dose after a previous chemotherapy regimen, and reported the proportion of patients getting FN. Meta-analysis was conducted to estimate the proportion of patients who experience one or more episodes of FN. The pooled, random effects meta-analysis estimate for the proportion of patients who experience one or more episodes of FN on docetaxel was 5.95% (95% CI 4.22-8.31) based on 13 studies, comprising 1609 patients. No significant differences were seen either between studies that permitted the use of prophylactic granulocyte colony-stimulating factors or between phase II and phase III trials.Evidence from randomised controlled trials suggests that the incidence of FN with docetaxel is around 6% and therefore an important factor to consider in the choice of the chemotherapy regimen. PMID- 19190634 TI - Reproductive factors and specific histological types of breast cancer: prospective study and meta-analysis. AB - Little is known about how reproductive factors affect the risk of breast cancers of different histology. In an analysis of prospective data on 1.2 million middle aged UK women, we used proportional hazards models to estimate the relative risks of six histological types in relation to menarche, childbearing and menopause. During 8.7 million person-years of follow-up, 17 923 ductal, 3332 lobular, 1062 tubular, 944 mixed ductal lobular, 330 mucinous and 117 medullary cancers were diagnosed. The effect of both age at menarche and age at first birth was greatest for lobular tumours; relative risks per 5-year increase in age at menarche for ductal, lobular, and tubular cancers were 0.93 (0.87-0.99), 0.65 (0.56-0.76), and 0.75 (0.57-0.98), respectively (P-value for heterogeneity=0.0001); and the relative risks per 5-year increase in age at first birth were 1.10 (1.07-1.12), 1.23 (1.17-1.29), and 1.13 (1.03-1.23), respectively (P-value for heterogeneity=0.0006). Increasing parity reduced the risk of each tumour type, except medullary cancers, but the reduction in risk was greater for mucinous cancers than for any other subtype considered (P<0.05 for comparison with each other subtype in turn). The effect of menopause did not vary significantly by tumour histology. Meta-analysis of published results on the effects of age at menarche and age at first birth on ductal and lobular cancers were in keeping with our findings. PMID- 19190635 TI - Dietary glycemic load and gastric cancer risk in Italy. AB - We investigated gastric cancer risk in relation to dietary glycemic index (GI) and glycemic load (GL), which represent indirect measures of carbohydrate absorption and consequently of dietary insulin demand, in a case-control study conducted in northern Italy between 1997 and 2007, including 230 patients with the incident, histologically confirmed gastric cancer and 547 frequency matched controls, admitted to the same hospitals as cases with acute non-neoplastic conditions. We used conditional logistic regression models, including terms for major recognised gastric cancer risk factors and non-carbohydrate energy intake. The odds ratios (ORs) in the highest vs lowest quintile were 1.9 (95% CI: 1.0 3.3) for GI and 2.5 (95% CI: 1.3-4.9) for GL. Compared with participants reporting low GL and high fruits/vegetables intake, the OR rose across strata of high GL and low fruits/vegetables, to reach 5.0 (95% CI: 2.2-11.5) for those reporting low fruits/vegetables intake and high GL. Our study may help to explain the direct relation observed in several studies between starchy foods and gastric cancer risk. PMID- 19190637 TI - Complex brain networks: graph theoretical analysis of structural and functional systems. AB - Recent developments in the quantitative analysis of complex networks, based largely on graph theory, have been rapidly translated to studies of brain network organization. The brain's structural and functional systems have features of complex networks--such as small-world topology, highly connected hubs and modularity--both at the whole-brain scale of human neuroimaging and at a cellular scale in non-human animals. In this article, we review studies investigating complex brain networks in diverse experimental modalities (including structural and functional MRI, diffusion tensor imaging, magnetoencephalography and electroencephalography in humans) and provide an accessible introduction to the basic principles of graph theory. We also highlight some of the technical challenges and key questions to be addressed by future developments in this rapidly moving field. PMID- 19190638 TI - The locus coeruleus and noradrenergic modulation of cognition. AB - Mood, attention and motivation co-vary with activity in the neuromodulatory systems of the brain to influence behaviour. These psychological states, mediated by neuromodulators, have a profound influence on the cognitive processes of attention, perception and, particularly, our ability to retrieve memories from the past and make new ones. Moreover, many psychiatric and neurodegenerative disorders are related to dysfunction of these neuromodulatory systems. Neurons of the brainstem nucleus locus coeruleus are the sole source of noradrenaline, a neuromodulator that has a key role in all of these forebrain activities. Elucidating the factors that control the activity of these neurons and the effect of noradrenaline in target regions is key to understanding how the brain allocates attention and apprehends the environment to select, store and retrieve information for generating adaptive behaviour. PMID- 19190639 TI - Dietary assessment at the end of life's spectrum. PMID- 19190640 TI - Cognitive processes in children's dietary recalls: insight from methodological studies. AB - BACKGROUND/OBJECTIVES: This article summarizes 12 dietary-reporting methodological studies with children (six validation studies, one non-validation study, five secondary analyses studies of data from one or more of the six validation studies), identifies research gaps and provides recommendations for (a) improving children's recall accuracy and (b) details to specify in publications of studies that utilize children's dietary recalls. SUBJECTS/METHODS: Randomly selected children (aged 9-10 years) were observed eating school breakfast and school lunch, and interviewed to obtain dietary recalls. RESULTS: Children's recall accuracy improved slightly between the first and third recalls, but an individual child's accuracy was inconsistent from one interview to the next. Although accuracy was poor overall, it was better for boys with reverse-order (evening-to-morning) prompts and for girls with forward-order (morning-to-evening) prompts. Children recalled breakfast intake less accurately than lunch intake. Children's accuracy did not depend on whether recalls were obtained in person or by telephone, but was better for recalls obtained with an open format than a meal format. Retention interval was crucial as children's accuracy was better for prior-24-h recalls (about the 24 h immediately preceding the interview) than for previous-day recalls (about midnight to midnight of the day before the interview). Observations of school meals did not affect children's recalls. Children's recall accuracy was related to their age/sex body mass index percentile. Conventional report rates (which disregard accuracy for items and amounts) overestimated accuracy for energy and macronutrients, and masked complexities of recall error. CONCLUSIONS: Research concerning errors in children's dietary recalls provides insight for improving children's recall accuracy. PMID- 19190641 TI - Diet and cognitive function in later life: a challenge for nutrition epidemiology. AB - BACKGROUND: As the proportion of old and very old people in the population increases, new research on the influence of diet on health and nutritional needs in later life will be needed. Dietary assessment methods that rely on short-term memory or lengthy interviews, such as the 24-h recall and diet history methods, could have some limitations in this age group. There is some support for the use of food frequency questionnaires (FFQs) in older people, although their validity in the older old and in those with more advanced cognitive decline has not been extensively assessed. SUBJECTS/METHODS: In a study designed to assess the validity of a modified FFQ in men and women over 65 years, 50 men and 47 women completed two FFQs, and 42 men and 41 women completed one FFQ followed by a 4-day weighed diet diary. Digit span forward (a test of short-term memory) and verbal fluency (a test of executive function) tests were used to assess the possible influence of cognitive function on repeatability and validity of the FFQ. RESULTS: The FFQ was found to have good repeatability for most nutrients and reasonable validity for some but not all nutrients. Cognitive function assessed prior to the dietary assessment showed no relationship with repeatability, but there was some evidence that validity was lower in those with lower executive function. CONCLUSIONS: Dietary assessment in healthy older people without overt cognitive decline can be achieved, but development and testing of methods of data collection for each target population and nutrient of interest are particularly important in this age group to ensure valid results. The possibility that cognitive decline influences dietary assessment needs to be borne in mind in the interpretation of observational studies of the influence of dietary intake on cognition in later life. PMID- 19190642 TI - Dietary assessment in the Avon Longitudinal Study of Parents and Children. AB - OBJECTIVES: To describe the use of dietary assessment methods in a cohort study. SUBJECTS: The Avon Longitudinal Study of Parents and Children (ALSPAC) based in Bristol, UK, a cohort of children born in 1991/1992 and their parents. METHODS AND RESULTS: Data were collected about maternal and paternal diet, breastfeeding, weaning and childhood diet by both food frequency questionnaire (FFQ) and unweighed diet records (children only). Other questionnaires have covered socioeconomic background and eating behaviours. Nutrient and food group intakes were calculated from both FFQ and diet records. These were used singly and in combination to assess the contribution of diet to various growth and development outcomes in the children and in relation to socioeconomic position and other behaviours. Both methods provided important contributions to knowledge and have enhanced each other, allowing the development of the assessment instruments as experience in their use was gained. Statistical analysis methods such as the use of dietary patterns to summarize the diet were explored. CONCLUSIONS: Longitudinal cohort studies are greatly enhanced if dietary assessment methods are built into their design. The use of more than one method of dietary assessment can increase the scientific usefulness of a study. PMID- 19190643 TI - Estimation of portion size in children's dietary assessment: lessons learnt. AB - BACKGROUND/OBJECTIVES: Assessing the dietary intake of young children is challenging. In any 1 day, children may have several carers responsible for providing them with their dietary requirements, and once children reach school age, traditional methods such as weighing all items consumed become impractical. As an alternative to weighed records, food portion size assessment tools are available to assist subjects in estimating the amounts of foods consumed. Existing food photographs designed for use with adults and based on adult portion sizes have been found to be inappropriate for use with children. This article presents a review and summary of a body of work carried out to improve the estimation of portion sizes consumed by children. METHODS: Feasibility work was undertaken to determine the accuracy and precision of three portion size assessment tools; food photographs, food models and a computer-based Interactive Portion Size Assessment System (IPSAS). These tools were based on portion sizes served to children during the National Diet and Nutrition Survey. As children often do not consume all of the food served to them, smaller portions were included in each tool for estimation of leftovers. The tools covered 22 foods, which children commonly consume. Children were served known amounts of each food and leftovers were recorded. They were then asked to estimate both the amount of food that they were served and the amount of any food leftover. RESULTS: Children were found to estimate food portion size with an accuracy approaching that of adults using both the food photographs and IPSAS. CONCLUSIONS: Further development is underway to increase the number of food photographs and to develop IPSAS to cover a much wider range of foods and to validate the use of these tools in a 'real life' setting. PMID- 19190644 TI - Dietary assessment at the end of life's spectrum. PMID- 19190645 TI - Use of technology in children's dietary assessment. AB - BACKGROUND: Information on dietary intake provides some of the most valuable insights for mounting intervention programmes for the prevention of chronic diseases. With the growing concern about adolescent overweight, the need to accurately measure diet becomes imperative. Assessment among adolescents is problematic as this group has irregular eating patterns and they have less enthusiasm for recording food intake. SUBJECTS/METHODS: We used qualitative and quantitative techniques among adolescents to assess their preferences for dietary assessment methods. RESULTS: Dietary assessment methods using technology, for example, a personal digital assistant (PDA) or a disposable camera, were preferred over the pen and paper food record. CONCLUSIONS: There was a strong preference for using methods that incorporate technology such as capturing images of food. This suggests that for adolescents, dietary methods that incorporate technology may improve cooperation and accuracy. Current computing technology includes higher resolution images, improved memory capacity and faster processors that allow small mobile devices to process information not previously possible. Our goal is to develop, implement and evaluate a mobile device (for example, PDA, mobile phone) food record that will translate to an accurate account of daily food and nutrient intake among adolescents. This mobile computing device will include digital images, a nutrient database and image analysis for identification and quantification of food consumption. Mobile computing devices provide a unique vehicle for collecting dietary information that reduces the burden on record keepers. Images of food can be marked with a variety of input methods that link the item for image processing and analysis to estimate the amount of food. Images before and after the foods are eaten can estimate the amount of food consumed. The initial stages and potential of this project will be described. PMID- 19190646 TI - Dietary assessment in early old age: experience from the Boyd Orr cohort. AB - BACKGROUND: The assessment of the impact of data quality issues, such as omitting to answer questions on a food frequency questionnaire (FFQ), is important in all study populations, including those in early old age. Assumptions about the limited nature of diets of older participants may influence the treatment and interpretation of their dietary data. SUBJECTS/METHODS: The Boyd Orr cohort is a long-term study based on 4999 UK men and women whose families took part in a survey of diet and health during 1937-1939. In 1997-1998, all 3182 traced, surviving study members, then aged 60 years and over, were sent a health and lifestyle questionnaire, including a 113-item FFQ, primarily to examine relationships between childhood and adult fruit, vegetable and antioxidant intakes. In-depth interviews were conducted with a purposively sampled subset of 31 respondents. RESULTS: Of the 1475 subjects who returned the questionnaire, 11% (n=161) had missing data on their FFQ. Those who omitted answers to more than 10 questions (n=127; 8.6%) were more likely to be aged over 70, to be female, but no more likely to report being overweight than those with 10 or fewer missing answers. Follow-up by telephone or post to reassess missing FFQ data was successful for 102 of the subjects with more than 10 omitted answers. Mean intakes of energy, fruit and vegetables, and selected nutrients were significantly increased after reassessment. The use of 'cross-check' questions to weigh fruit and vegetable intake (n=1383) showed potentially systematic errors in the reporting of these foods, vitamin C and carotene. Analysis of interview data among a subset of participants partially challenged stereotypical views of the diets of older people with, for example, increased freedom in food choice associated with life transitions. CONCLUSIONS: Food frequency questionnaires for those in early old age, as for others, need to meet competing demands of being comprehensive for those with varied diets, while not being so onerous that they deter completion. Reviewing questionnaires with participants remains important in this group, as omitting to answer questions on the FFQ does not necessarily equate to non-consumption. Qualitative interviews may aid in the interpretation of the quantitative data obtained. PMID- 19190647 TI - Nutrition in advanced age: dietary assessment in the Newcastle 85+ study. AB - BACKGROUND/OBJECTIVES: Assessing food choice and/or nutrient intake in older people, particularly the oldest old (85 years and over), presents particular challenges. In some cases the respondent may have little or no involvement in food acquisition or preparation, in others, cognitive/memory impairment may restrict the ability to recall intake, or physical limitations may affect the ability to record intake. The assessment may therefore need to involve whoever provides care for the older person, of whom there may be more than one. For these reasons, there is a need for validated methods for dietary assessment in large populations within this age range. The need is particularly acute in view of the secular increase in the numbers of older people and the interest in the role of nutrition in maintaining health and ameliorating age-related decline. This paper describes a comparison of two different methods of dietary assessment within the Newcastle 85+ Study; a UK cohort study of health and ageing in the oldest old. METHODS: Two methods, the food frequency questionnaire (FFQ) (based on broad recall of the previous 12 months intake) and the repeated multiple pass recall (MPR) tool (based on detailed recall of the previous day's intake on two separate occasions), were applied in two different groups of approximately 85 individuals aged 85 years. FFQ data were collected during a pilot study conducted between 2003 and 2004, MPR data were collected in the main Newcastle study in 2006. Relative validity was measured by calculation of the ratio of reported energy intake to estimated basal metabolic rate (EI/BMR) and by comparison with dietary intakes reported for subjects of similar age in the UK National Diet and Nutrition Survey. RESULTS: EI/BMR ratios for MPR were 1.56 and 1.39 for men and women, respectively, and for FFQ were 2.18 and 2.14. The FFQ was found to overestimate energy and nutrient intake considerably. The MPR gave more realistic estimates of energy and nutrient intakes, and was found to be acceptable for use in this population group. However, use of this tool required greater investigator (nurse) time, extra resources for training and quality assurance and additional time and expertise in data processing. CONCLUSIONS: In the Newcastle 85+ Study, where the overall aims include detailed investigation of diet in relation to many variables describing biological, clinical and psychosocial status, we concluded that MPR was the preferable method, although there remains a need for non subjective methods for assessing dietary intake, that is, biomarker approaches, which can give a comprehensive and objective assessment of dietary exposure. PMID- 19190648 TI - Experience of dietary assessment and validation from three Swedish studies in the elderly. AB - BACKGROUND/OBJECTIVES: The elderly are a growing part of the population. The objectives have been to describe the dietary assessment methods undertaken, and to evaluate the different validation methods and their performance in free-living elderly populations. SUBJECTS/METHODS: Participants were from three studies, the Johanneberg, H70 and Nordic Research on Ageing (NORA) studies undertaken in Sweden. The mean age across these studies ranged from 70 to 80 years, and cohort size ranged from n=173 to a total of 1360 in the four cohorts of the H70 geriatric population study. A food frequency questionnaire (FFQ) was used in Johanneberg and a diet history (DH) in H70 and NORA. Validation was undertaken using a 4-day food record (FR), four 24-h urinary nitrogen (UN) collections and energy intake/basal metabolic rate (EI/BMR) cutoffs in Johanneberg, EI/BMR cutoffs, total energy expenditure (TEE) estimated by heart rate monitoring, activity diary and doubly labelled water in H70, and EI/BMR cutoffs in NORA. RESULTS: The FFQ provided higher intakes of EI and nutrients vs FR, confirmed by UN for protein. Compared with EI/BMR cutoffs, overestimation of EI was moderate. DH showed that slight underestimation vs heart rate was in accordance with activity diary, but compared with doubly labelled water, DH underestimated EI by 12%. In NORA, EI/BMR was high compared with EI/BMR cutoffs. Validation methods detected under- and over-reporting. EI and nutrient intake were in accordance with recommendations. CONCLUSIONS: Elderly up to their 80s can perform well in dietary studies. However, people over the age of 80 years may tend to report food habits from earlier in life. PMID- 19190649 TI - Dietary assessment in elderly people: experiences gained from studies in the Netherlands. AB - BACKGROUND/OBJECTIVES: In selecting a dietary assessment method, several aspects such as the aim of the study and the characteristics of the target population should be taken into account. In elderly people, diminished functionality and cognitive decline may hamper dietary assessment and require tailored approaches to assess dietary intake. The objective of this paper is to summarize our experience in dietary assessment in a number of different studies in population groups over 65 years of age in the Netherlands, and to discuss this experience in the perspective of other nutrition surveys in the elderly. METHODS: In longitudinal studies, we applied a modified dietary history; in clinical nursing home studies, trained staff observed and recorded food consumption; and in a controlled trial in healthy elderly men, we used a food frequency questionnaire (FFQ). RESULTS: For all methods applied in the community-dwelling elderly people, validation studies showed a similar underestimation of intake of 10-15% compared with the reference value. In the care-depending elderly, the underestimation was less: 5% according to an observational method. The methods varied widely in the resources required, including burden to the participants, field staff and finances. CONCLUSIONS: For effective dietary assessment in older adults, the major challenge will be to distinguish between those elderly who are able to respond correctly to the less intensive methods, such as 24-h recalls or FFQ, and those who are not able to respond to these methods and require adapted techniques, for example, observational records. PMID- 19190650 TI - Workshop 1: Use of technology in dietary assessment. PMID- 19190651 TI - Workshop 2: The use of surrogate reporters in the assessment of dietary intake. PMID- 19190652 TI - Workshop 3: Novel approaches for estimating portion sizes. PMID- 19190653 TI - Inhibition of poly(ADP-ribose) polymerase suppresses inflammation and promotes recovery after ischemic injury. AB - The brain inflammatory response induced by stroke contributes to cell death and impairs neurogenesis. Poly(ADP-ribose) polymerase-1 (PARP-1) is a coactivator of the transcription factor NF-kappaB and required for NF-kappaB-mediated inflammatory responses. Here we evaluated PARP inhibition as a means of suppressing post-stroke inflammation and improving outcome after stroke. Rats were subjected to bilateral carotid occlusion-reperfusion, and treatment with the PARP inhibitor N-(6-oxo-5,6-dihydrophenanthridin-2-yl)-N,N-dimethylacetamide (PJ34) was begun 48 h later. PJ34 was found to rapidly suppress the ischemia induced microglial activation and astrogliosis. Behavioral tests performed 6 to 8 weeks after ischemia showed deficits in spatial memory and learning that were lessened by the PJ34 treatment. Immunohistochemical evaluation of hippocampus at 8 weeks after ischemia showed increased neuronal density in CA1 layer of PJ34 treated animals relative to vehicle-treated animals. Bromodeoxyuridine labeling showed formation of new neurons in hippocampal CA1 area in PJ34-treated animals, but not in vehicle-treated animals. Together, these results suggest that treatment with a PARP inhibitor for several days after ischemia enhances long term neuronal survival and neurogenesis by reducing inflammation. PMID- 19190654 TI - Sex differences in minocycline-induced neuroprotection after experimental stroke. AB - Minocycline is neuroprotective in clinical and experimental stroke studies, due in part to its ability to inhibit poly (ADP-ribose) polymerase. Previous preclinical data have shown that interference with poly (ADP-ribose) polymerase signaling leads to sex-specific neuroprotection, reducing stroke injury only in males. In this study, we show that minocycline is ineffective at reducing ischemic damage in females after middle cerebral artery occlusion, likely due to effects on poly (ADP-ribose) polymerase signaling. Clinical trials must consider possible sex differences in the response to neuroprotective agents, if we hope to translate promising therapies to stroke patients of both sexes. PMID- 19190655 TI - In vivo evidence for cerebral depletion in high-energy phosphates in progressive supranuclear palsy. AB - Indirect evidence from laboratory studies suggests that mitochondrial energy metabolism is impaired in progressive supranuclear palsy (PSP), but brain energy metabolism has not yet been studied directly in vivo in a comprehensive manner in patients. We have used combined phosphorus and proton magnetic resonance spectroscopy to measure adenosine-triphosphate (ATP), adenosine-diphosphate (ADP), phosphorylated creatine, unphosphorylated creatine, inorganic phosphate and lactate in the basal ganglia and the frontal and occipital lobes of clinically probable patients (N=21; PSP stages II to III) and healthy controls (N=9). In the basal ganglia, which are severely affected creatine in PSP patients, the concentrations of high-energy phosphates (=ATP+phosphorylated creatine) and inorganic phosphate, but not low-energy phosphates (=ADP+unphosphorylated creatine), were decreased. The decrease probably does not reflect neuronal death, as the neuronal marker N-acetylaspartate was not yet significantly reduced in the early-stage patients examined. The frontal lobe, also prone to neurodegeneration in PSP, showed similar alterations, whereas the occipital lobe, typically unaffected, showed less pronounced alterations. The levels of lactate, a product of anaerobic glycolysis, were elevated in 35% of the patients. The observed changes in the levels of cerebral energy metabolites in PSP are consistent with a functionally relevant impairment of oxidative phosphorylation. PMID- 19190657 TI - Protective effects of renin-angiotensin blockade beyond blood pressure control. AB - Antihypertensive drugs exert a number of blood pressure-independent benefits. However, demonstrating the clinical significance of these effects may be difficult for a number of reasons. First, blood pressure can be measured in the clinic, at home and over the 24-h period by ambulatory monitoring. Second, differences between these measures mean that achieving equivalent blood pressure reductions in two treatment arms may be difficult, if not impossible. Furthermore, even small differences in blood pressure can translate into significant effects on cardiovascular risk, especially in the later stages of the cardiovascular continuum. In large clinical trials, other errors limiting the sensitivity to treatment differences include high patient dropouts and unplanned crossover. In addition, as so many patients fail to achieve blood pressure goals even in clinical trials where patient's and physician's motivation is high, the need for cardiovascular protection beyond blood pressure control is unequivocal. Clinical trials of angiotensin II receptor blockers have suggested significant effects beyond blood pressure control, which are observed throughout and with greater consistency in the early phases of the cardiovascular continuum. There may also be differences between angiotensin II receptor blockers and angiotensin converting enzyme inhibitors. Conclusive demonstration that these blood pressure independent effects do exist will require, however, a much more accurate and extended assessment of the blood pressure effects of the drugs. PMID- 19190656 TI - Haemostatic markers are associated with measures of vascular disease in adults with hypertension. AB - Haemostatic markers have been implicated in the development and progression of vascular disease. We investigated the associations of several haemostatic markers (fibrinogen, D-dimer, FV, FVII, FVIII, von Willebrand factor (vWF) and antithrombin III) with two quantitative measures of vascular disease in adults with hypertension. Participants included 1051 African Americans (65+/-9 years, 72% women) and 894 non-Hispanic whites (61+/-9 years, 55% women) belonging to hypertensive sibships. Phenotypes of vascular disease included the ankle-brachial index (ABI), a measure of peripheral arterial disease, and urinary albumin/creatinine ratio (UACR), a surrogate of glomerular endothelial function. Generalized estimating equations were used to assess whether plasma levels of haemostatic markers were associated with measures of arteriosclerosis, after adjustment for conventional risk factors and medication (statin, aspirin and oestrogen) use. Higher fibrinogen and D-dimer were significantly associated with lower ABI in African Americans (P<0.001 and 0.004 respectively) and in non Hispanic whites (P<0.001 and 0.010 respectively). Higher fibrinogen (P<0.001), D dimer (P=0.003), FVIII (P<0.001) and vWF (P<0.001) were significantly associated with higher UACR in African Americans, whereas, in non-Hispanic whites, higher fibrinogen (P=0.020) and FVII (P=0.006) were significantly associated with higher UACR. Our findings indicate that in adults with essential hypertension, several markers in the haemostatic pathway are independently associated with ABI and UACR, two measures of vascular disease.. PMID- 19190658 TI - Efficacy and safety of initial combination therapy with amlodipine/valsartan compared with amlodipine monotherapy in black patients with stage 2 hypertension: the EX-STAND study. AB - The strategy of initiating hypertension treatment with combination versus single drug therapy was formally tested in a prospective, double-blind, parallel-group trial in blacks with stage 2 hypertension (mean sitting systolic BP (MSSBP) >or=160 and <200 mm Hg). Participants were randomized equally to amlodipine/valsartan (A/V) (n=286) or amlodipine (A) monotherapy (n=286). After 2 weeks, there was forced titration of A/V 5/160 mg to A/V 10/160 mg and of A 5 to A 10 mg followed by 10 additional weeks of treatment. If SBP was >or=130 mm Hg at week 4, the protocol allowed optional titration of A/V to the 10/320 mg dose and, at week 8, hydrochlorothiazide 12.5 mg was optionally added to both A/V and A if SBP >or=130 mm Hg. Amlodipine/valsartan at week 8 lowered MSSBP last observation carried forward significantly>A (33.3 vs 26.6 mm Hg, P<0.0001). Lowering of MSSBP with A/V significantly exceeded that of A in several specified subgroups-the elderly (>or=65 years), isolated systolic hypertension, and those with body mass index (BMI) >or=30 kg/m(2). More patients treated with A/V than A achieved BP control (<140/90 mm Hg) both at weeks 8 (49.8 vs 30.2%; P<0.0001) and 12 (57.2 vs 35.9%; P<0.0001). Both treatment regimens were well tolerated. In conclusion, the strategy of initiating combination antihypertensive drug therapy in blacks with stage 2 hypertension with amlodipine /valsartan achieves greater and quicker reductions in BP as well as significantly higher BP control rates than starting treatment with amlodipine monotherapy. PMID- 19190659 TI - The year that was and the year ahead. AB - Now that we are well into 2009, I can't help but think about the year that has passed. Fear not, this will not be one of those dreaded holiday letters where we list all the highs and lows of the year. But as I look back, there are many things I hope that I have permanently crossed off my "To Do" list and others that I am looking forward to doing. PMID- 19190660 TI - Tip20p reaches out to Dsl1p to tether membranes. PMID- 19190661 TI - Wedging out DNA damage. PMID- 19190662 TI - Towards the architecture of the chromosomal architects. PMID- 19190665 TI - Complications of transpyloric nutrition in children. PMID- 19190664 TI - Nonsense-mediated mRNA decay (NMD) mechanisms. AB - Nonsense-mediated mRNA decay (NMD) is a translation-coupled mechanism that eliminates mRNAs containing premature translation-termination codons (PTCs). In mammalian cells, NMD is also linked to pre-mRNA splicing, as in many instances strong mRNA reduction occurs only when the PTC is located upstream of an intron. It is proposed that in these systems, the exon junction complex (EJC) mediates the link between splicing and NMD. Recent studies have questioned the role of splicing and the EJC in initiating NMD. Instead, they put forward a general and evolutionarily conserved mechanism in which the main regulator of NMD is the distance between a PTC and the poly(A) tail of an mRNA. Here we discuss the limitations of the new NMD model and the EJC concept; we argue that neither satisfactorily accounts for all of the available data and offer a new model to test in future studies. PMID- 19190667 TI - Longitudinal changes in energy expenditure in an elderly German population: a 12 year follow-up. AB - BACKGROUND/OBJECTIVES: This study investigates age-dependent changes in different components of energy expenditure (EE) within the longitudinal study on nutrition and health status in an aging population in Giessen, Germany (GISELA). SUBJECTS/METHODS: Between 1994 and 2006, data obtained at a total of 3033 visits from 363 women and 153 men with a mean initial age of 67.4+/-5.9 and 66.9+/-5.2 years, respectively, were evaluated. The mean duration of follow-up was 8 years. Resting metabolic rate (RMR) was assessed by indirect calorimetry and physical activity patterns were assessed by questionnaire. EE of physical activity and total EE (TEE) were calculated using multipliers for RMR. Energy intake was determined through a validated 3-day estimated dietary record. Linear mixed models were used to analyze the influence of age on EE adjusted for covariates. RESULTS: Resting metabolic rate decreased in women and men by 158 and 326 kJ/d per decade, respectively; after considering changes in body composition and fat distribution, respective decreases were 81 and 286 kJ/d per decade. EE of physical activity decreased similarly in both sexes (472 kJ/d per decade). TEE dropped in women and men by 540 and 823 kJ/d per decade, respectively. No statistically significant changes in energy intake and body weight were observed in the course of follow-up. CONCLUSIONS: The age-dependent decrease in TEE is mainly due to a decrease in physical activity. The stable energy intake and body weight of the GISELA subjects may be indicators for a relatively good health status. PMID- 19190668 TI - Zinc status and cognitive function of pregnant women in Southern Ethiopia. AB - The relation between zinc status and cognitive function was examined in a cross sectional study in the Sidama area of Southern Ethiopia. Pregnant women >24 weeks of gestation from three adjacent rural villages volunteered to participate. Mean (s.d.) plasma zinc of 99 women was 6.97 (1.07) mumol/l (below the cutoff of 7.6 mumol/l indicative of zinc deficiency at this stage of gestation). The Raven's Coloured Progressive Matrices (CPM) test was administered individually. Scores for the Raven's scale A, which is the simplest scale, ranged from 4 to 10 of a possible 12. Women with plasma zinc <7.6 mumol/l had significantly lower Raven's CPM scale A scores than women with plasma zinc concentrations >7.6 mumol/l. Plasma zinc and maternal age and education predicted 17% of the variation in Raven's CPM scale A scores. We conclude that zinc deficiency is a major factor affecting cognition in these pregnant women. PMID- 19190669 TI - Feminization and the collapse of haplodiploidy in an asexual parasitoid wasp harboring the bacterial symbiont Cardinium. AB - Cardinium is a bacterial symbiont infecting many species of arthropods, and is associated with manipulation of host reproduction. Cardinium is the causal agent of asexual reproduction, or thelytoky, in the chalcidoid parasitoid wasp Encarsia hispida. Feeding antibiotics to the infected adult females results in uninfected male offspring. Here, we show that these males are diploid. Diploid males are extremely unusual in the large hymenopteran superfamily Chalcidoidea, and, to our knowledge, have never before been associated with symbiont infection in this group. These findings indicate that at least in E. hispida, diploidy restoration is a necessary condition but not sufficient to elicit female development. Cardinium is required to feminize diploid male embryos and thus must interact with elements of the host sex determination system. In addition, our data suggest that Cardinium is necessary for the fertility of E. hispida; antibiotic curing of Cardinium reduces offspring production of adult females. PMID- 19190670 TI - The initial steps of myofibril assembly: integrins pave the way. AB - Myofibril assembly results in a regular array of identical sarcomeres in striated muscle. Sarcomere structure is conserved across the animal kingdom, which implies that the mechanisms of myofibril assembly are also likely to be conserved. Recent advances from model genetic systems and insights from stress fibre cell biology have shed light on the mechanisms that set sarcomere spacing and the initial assembly of sarcomere arrays. We propose a model of integrin-dependent cell matrix adhesion as the starting point for myofibrillogenesis. PMID- 19190672 TI - A novel VPS13B mutation in two brothers with Cohen syndrome, cutis verticis gyrata and sensorineural deafness. AB - We have earlier described a syndrome characterized by microcephaly, cutis verticis gyrata, retinitis pigmentosa, cataracts, hearing loss and mental retardation (Mendelian inheritance in man (MIM) no: 605685) in two brothers from a non-consanguineous Lebanese family. In view of the rarity of the disorder and the high rate of inbreeding in the Lebanese population, we assumed an autosomal recessive trait inherited from a common ancestor. A genomewide scan was performed. The single locus on the long arm of chromosome 8 that showed homozygosity by descent comprised the gene responsible for Cohen syndrome (CS), VPS13B. We then sequenced VPS13B in the patients and found a homozygous splice site mutation. Several possible explanations for the overlap between CS and the clinical features observed in our patients are discussed. Our data highlight the potential of high-resolution homozygosity mapping in small populations with a high rate of inbreeding. PMID- 19190671 TI - Single proteins might have dual but related functions in intracellular and extracellular microenvironments. AB - The maintenance of organ homeostasis and the control of an appropriate response to environmental alterations require the intimate coordination of cellular functions and tissue organization. An important component of this coordination could be provided by proteins that can have distinct but linked functions on both sides of the plasma membrane. We present a model that proposes that unconventional secretion provides a mechanism through which single proteins can integrate complex tissue functions. PMID- 19190673 TI - Increased lung cancer risks are similar whether arsenic is ingested or inhaled. AB - In 1980, the International Agency for Research on Cancer (IARC) determined there was sufficient evidence to support that inorganic arsenic was a human lung carcinogen based on studies involving exposure through inhalation. In 2004, IARC listed arsenic in drinking water as a cause of lung cancer, making arsenic the first substance established to cause human cancer through two unrelated pathways of exposure. It may initially seem counterintuitive that arsenic in drinking water would cause human lung cancer, and even if it did, one might expect risks to be orders of magnitude lower than those from direct inhalation into the lungs. In this paper, we consider lung cancer dose-response relationships for inhalation and ingestion of arsenic by focusing on two key studies, a cohort mortality study in the United States involving Tacoma smelter workers inhaling arsenic, and a lung cancer case-control study involving ingestion of arsenic in drinking water in northern Chile. When exposure was assessed based on the absorbed dose identified by concentrations of arsenic in urine, there was very little difference in the dose-response findings for lung cancer relative risks between inhalation and ingestion. The lung cancer mortality rate ratio estimate was 8.0 (95% CI 3.2-16.5, P<0.001) for an average urine concentration of 1179 microg/l after inhalation, and the odds ratio estimate of the lung cancer incidence rate ratio was 7.1 (95% CI 3.4-14.8, P<0.001) for an estimated average urine concentration of 825 microg/l following ingestion. The slopes of the linear dose response relationships between excess relative risk (RR-1) for lung cancer and urinary arsenic concentration were similar for the two routes of exposure. We conclude that lung cancer risks probably depend on absorbed dose, and not on whether inorganic arsenic is ingested or inhaled. PMID- 19190674 TI - The effect of joint exposures: examining the presence of interaction. AB - Clinical epidemiological studies investigate whether an exposure, or risk factor, is causally related to the development or progression of a disease or mortality. It might be of interest to study whether this relation is different in different types of patients. To address such research questions, the presence of interaction among risk factors can be examined. Causal interaction between two risk factors is considered most clinically relevant in epidemiology. Causal interaction occurs when two risk factors act together in causing disease and is explicitly defined as a deviation from additivity on a risk difference scale. Statistical interaction can be evaluated on both an additive (absolute risk) and multiplicative (relative risk) scale, depending on the model that is used. When using logistic regression models, which are multiplicative models, several measures of additive interaction are presented to evaluate whether the magnitude of an association differs across subgroups: the relative excess risk due to interaction (RERI), the attributable proportion due to interaction (AP), or the synergy index (S). For a transparent presentation of interaction effects the recent Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) Statement advises reporting the separate effect of each exposure as well as the joint effect compared with the unexposed group as a joint reference category to permit evaluation of both additive and multiplicative interaction. PMID- 19190675 TI - Latest findings in phosphate homeostasis. AB - The kidney is a key player in phosphate balance. Inappropriate renal phosphate transport may alter serum phosphate concentration and bone mineralization, and increase the risk of renal lithiasis or soft tissue calcifications. The recent identification of fibroblast growth factor 23 (FGF23) as a hormone regulating phosphate and calcitriol metabolism and of klotho has changed the understanding of phosphate homeostasis; and a bone-kidney axis has emerged. In this review, we present recent findings regarding the consequences of mutations affecting several human genes encoding renal phosphate transporters or proteins regulating phosphate transport activity. We also describe the role played by the FGF23 klotho axis in phosphate homeostasis and its involvement in the pathophysiology of phosphate disturbances in chronic kidney disease. PMID- 19190677 TI - Fully phosphorylated fetuin-A forms a mineral complex in the serum of rats with adenine-induced renal failure. AB - The serum glycoprotein fetuin-A is an important inhibitor of extra-osseous calcification, but correlations between serum fetuin-A levels and the extent of vascular calcification are controversial. In this study, we used a rat model of adenine-induced renal failure with secondary hyperparathyroidism that exhibits all characteristic features of patients with chronic kidney disease. These rats had medial vascular calcification along with reduced levels of both serum and hepatic fetuin-A. Treatment with an inhibitor of ectopic calcification, alendronate, decreased bone turnover and eliminated completely the vascular calcification in this rat model, but there was no change in the levels of hepatic and serum fetuin-A. Centrifugation of the serum of untreated rats with renal failure gave a small precipitate composed of fetuin-A, calcium, magnesium, and phosphate; this complex, absent from normal rat serum, was not found in the serum of alendronate-treated rats with renal failure. Rat serum contained three types of phosphorylated fetuin-A, as well as unphosphorylated forms, but only the fully phosphorylated fetuin-A was present in the mineral complex. The amount of this complex reflected the risk of mineral precipitation. Our results suggest that the measurement of serum fetuin-mineral complex rather than fetuin-A alone might provide a better indication of extra-osseous calcification propensity. PMID- 19190678 TI - Increased tissue acid mediates a progressive decline in the glomerular filtration rate of animals with reduced nephron mass. AB - The combination of an acid-inducing diet and reduced nephron mass is associated with a progressive decline in glomerular filtration rate (GFR) that can be corrected by dietary alkali. Here we determined whether the higher tissue acid content mediates the decline in GFR. Using Munich-Wistar rats we induced sub total nephrectomy and measured by microdialysis the tissue acid content in the kidney cortex and in the paraspinous muscle. The GFR was lower in the rats with reduced nephron mass at 1 and 13 weeks following subtotal nephrectomy compared to the sham-operated rats. Both groups of rats ate the same acid-inducing casein based diet and had similar plasma acid-base parameters and net urine acid excretion. However, rats with reduced nephron mass had higher tissue acid content compared to control animals and had a lower GFR at week 13 compared to that measured at week 1. Adding dietary acid to the casein diet led to an even higher tissue acid and lower GFR by week 13. By contrast, adding alkali to the casein diet or placing animals with reduced nephron mass on a soy-based diet led to a lower tissue acid content and no decline in GFR. Animals with reduced nephron mass on a soy-based diet given dietary acid had a higher tissue acid content and a decline in GFR. These studies show that dietary maneuvers that increase the tissue acid content reduce GFR, whereas diets that lower the tissue acid level preserve GFR during chronic kidney failure. PMID- 19190676 TI - The kallikrein-kinin system in health and in diseases of the kidney. AB - Since kallikrein was discovered as a vasodilatory substance in human urine, the kallikrein-kinin system (KKS) has been considered to play a physiological role in controlling blood pressure. Gene targeting experiments in mice in which the KKS has been inactivated to varying degrees have, however, questioned this role, because basal blood pressures are not altered. Rather, these experiments have shown that the KKS has a different and important role in preventing changes associated with normal senescence in mice, and in reducing the nephropathy and accelerated senescence-associated phenotypes induced in mice by diabetes. Other experiments have shown that the KKS suppresses mitochondrial respiration, partly by nitric oxide and prostaglandins, and that this suppression may be a key to understanding how the KKS influences senescence-related diseases. Here we review the logical progression and experimental data leading to these conclusions, and discuss their relevance to human conditions. PMID- 19190679 TI - Thiazolidinedione use is associated with better survival in hemodialysis patients with non-insulin dependent diabetes. AB - Cardiovascular mortality is especially high among dialysis patients with diabetes, as is morbidity due to protein energy wasting. Given that both of these factors may be decreased by thiazolidinedione treatment, we studied the effect of thiazolidinedione use on survival among chronic dialysis patients in a national cohort of 5290 incident dialysis patients with diabetes. Thiazolidinedione use was assessed according to prescription data, and the analyses were stratified based on insulin use due to observed interaction. In the primary analysis, thiazolidinedione treatment was associated with significantly lower all-cause mortality among insulin-free but not insulin-requiring subjects, with adjusted hazards ratios of 0.53 (0.31-0.89) and 0.82 (0.46-1.47) respectively. Sensitivity analyses found the findings to be robust with respect to confounding by indication, severity of the diabetes, potential reverse causality, and time varying exposure patterns. The mechanism of this decline in all-cause mortality will need to be examined after these studies are confirmed. PMID- 19190681 TI - Mechanisms of tubular volume retention in immune-mediated glomerulonephritis. AB - Glomerulonephritis is characterized by hematuria, proteinuria, hypertension, and edema, but the mechanisms contributing to volume disorders are controversial. Here we used the rat anti-Thy1 model of mesangioproliferative glomerulonephritis to test the hypothesis that disturbed salt and water homeostasis is based on tubular epithelial changes that cause salt retention. In this model there was an early onset of pronounced proteinuria and lipiduria associated with reduced fractional sodium excretion and a lowering of the renin-angiotensin-aldosterone system. The glomerular filtration rate and creatinine clearance were decreased on day 6. There was a reduced abundance of the major salt and water transport proteins on the proximal tubular brush border membrane and which paralleled cellular protein overload, enhanced membrane cholesterol uptake and cytoskeletal changes. Alterations in thick ascending limb were moderate. Changes in the collecting ducts were characterized by an enhanced abundance and increased subunit cleavage of the epithelial sodium channel, both events consistent with increased sodium reabsorption. We suggest that irrespective of the proximal tubular changes, altered collecting duct sodium reabsorption may be crucial for volume retention in acute glomerulonephritis. We suggest that enhanced proteolytic cleavage of ion transporter subunits might be a novel mechanism of channel activation in glomerular diseases. Whether these proteases are filtered or locally secreted awaits determination. PMID- 19190680 TI - Kidney-specific reconstitution of the A1 adenosine receptor in A1 adenosine receptor knockout mice reduces renal ischemia-reperfusion injury. AB - Genetic deletion of the adenosine A1 receptor (A1AR) increased renal injury following ischemia-reperfusion injury suggesting that receptor activation is protective in vivo. Here we tested this hypothesis by expressing the human-A(1)AR in A(1)AR knockout mice. Renal ischemia-reperfusion was induced in knockout mice 2 days after intrarenal injection of saline or a lentivirus encoding enhanced green fluorescent protein (EGFP) or EGFP-human-A(1)AR. We found that the latter procedure induced a robust expression of the reporter protein in the kidneys of knockout mice. Mice with kidney-specific human-A(1)AR reconstitution had significantly lower plasma creatinine, tubular necrosis, apoptosis, and tubular inflammation as evidenced by decreased leukocyte infiltration, pro-inflammatory cytokine, and intercellular adhesion molecule-1 expression in the kidney following injury compared to mice injected with saline or the control lentivirus. Additionally, there were marked disruptions of the proximal tubule epithelial filamentous (F)-actin cytoskeleton in both sets of control mice upon renal injury, whereas the reconstituted mice had better preservation of the renal tubule actin cytoskeleton, which co-localized with the human-A(1)ARs. Consistent with reduced renal injury, there was a significant increase in heat shock protein 27 expression, also co-localizing with the preserved F-actin cytoskeleton. Our findings suggest that selective expression of cytoprotective A(1)ARs in the kidney can attenuate renal injury. PMID- 19190682 TI - Central administration of Orphanin FQ inhibits GnRH secretion by ORL1 receptor in the median eminence of freely moving ovariectomized rats. AB - OBJECTIVE: This study aimed to investigate the possible role of Orphanin FQ (OFQ) in the regulation of hypo-thalamic gonadotropin-releasing hormone (GnRH) secretion. METHODS: The method of push-pull perfusion and radioimmuno-assay (RIA) were adopted to examine the secretory profile of GnRH in the median eminence (ME) in freely moving ovari-ectomized (OVX) rats after intracerebroventricular (icv) injection of OFQ and/or [Nphe(1)]NC(1-13)NH(2) (NC13), a competitive antagonists of the opioid receptor-like 1 receptor (ORL1 receptor). RESULTS: GnRH release from ME significantly decreased from 40 min to 80 min after the administration of 20 and 200 nmol OFQ in OVX rats (P < 0.05). This inhibitory effect of 20 nmol OFQ could be abolished by pretreatment with equal dose of NC13. More interestingly, GnRH secretion from ME was increased markedly 60 min after icv injection of 100 and 200 nmol NC13 (P < 0.05). CONCLUSION: Our results suggested central administration of OFQ could inhibit the release of GnRH in the ME of hypothalamus through ORL1 receptor, providing further in vivo evidence supporting the role of OFQ in the control of GnRH secretion. PMID- 19190685 TI - Initial bradykinin triggers calcium-induced calcium release in C6 glioma cells and its significance. AB - OBJECTIVE: To investigate the underlying mechanism for the selective modulation of the permeability of blood-tumor barrier (BTB) by small dose of bradykinin (BK). METHODS: C6 glioma cells were treated with BK, and changes of intracellular nitric oxide (NO) and intracellular calcium level were measured with fluorescent spectrophotometer. RESULTS: The initial application of BK easily triggered extracellular calcium influx, which resulted in intracellular calcium store release in C6 glioma cells. The above mechanism was also named ryanodine mediated calcium induced calcium release (CICR). We also detected a long-lasting intracellular NO elevation in C6 glioma cells upon BK treatment. Further study showed that ryanodine mediated CICR contributed greatly to the secondary NO elevation induced by BK treatment. CONCLUSION: These results suggested that BK triggered CICR in C6 glioma cells and the associated NO generation might be the underlying mechanism for the selective modulation of BTB permeability by BK. PMID- 19190683 TI - Pro-protein convertase-2/carboxypeptidase-E mediated neuropeptide processing of RGC-5 cell after in vitro ischemia. AB - OBJECTIVE: To observe the change of the neuropeptide pro-protein processing system in the ischemic retina ganglion cell-5 (RGC-5) cells, pro-protein convertase-2 (PC2), carboxypeptidase-E (CPE) and preproneuropeptide Y (preproNPY) protein levels in the ischemic RGC-5 cells and conditioned medium were analyzed. METHODS: The RGC-5 cell was differentiated in 0.1 mumol/L staurosporine for 24 h and then stressed by different doses of oxygen and glucose deprivation (OGD). The acute or chronic OGD-induced cell death rates were obtained by using PI or TUNEL staining. The protein expression levels were determined by using the Western blot method and PC2 activity analysis. RESULTS: The ischemia caused substantial cell death in an OGD dose-dependent manner. In the cells, proPC2 and preproNPY protein levels gradually increased whereas proCPE gradually decreased. After OGD, PC2 activity was decreased. In the conditioned medium, proPC2 and PC2 proteins gradually decreased whereas proCPE, CPE, and preproNPY proteins gradually increased. CONCLUSION: These results demonstrated that OGD inhibited the neuropeptide pro-protein processing system by reducing PC2 activity and the maturation of proPC2. The aggregation of the pro-proteins and the increase of the active CPE excision adversely exacerbated the cell injury. The pro-protein processing system might play a critical role in the ischemic stress of RGC-5 cells. PMID- 19190684 TI - Firing activity of locus coeruleus noradrenergic neurons increases in a rodent model of Parkinsonism. AB - OBJECTIVE: To investigate the changes in the firing activity of noradrenergic neurons in the locus coeruleus (LC) in a rat model of Parkinson disease (PD). METHODS: 2 and 4 weeks after unilateral lesion of the nigrostriatal pathway in the rat by local injection of 6-hydroxydopamine (6-OHDA) into the right substantia nigra pars compacta (SNc), the firing activity of noradrenergic neurons in LC was recorded by extracellular single unit recording. RESULTS: The firing rate of LC noradrenergic neurons increased significantly 2 and 4 weeks after 6-OHDA lesions compared to normal rats, respectively (P < 0.05). The percentage of irregularly firing neurons was obviously higher than that of normal rats during the fourth week after SNc lesion (P < 0.05). CONCLUSION: LC noradrenergic neurons are overactive and more irregular in 6-OHDA-lesioned rats. These changes suggest an implication of the LC in the pathophysiological mechanism of PD. PMID- 19190686 TI - Therapeutic potential of extracellular ATP and P2 receptors in nervous system diseases. AB - Extracellular adenosine 5 inch-triphosphate (ATP) is a key signaling molecule present in the central nervous system (CNS), and now is receiving greater attention due to its role as a messenger in the CNS during different physiological and pathological events. ATP is released into the extracellular space through vesicular exocytosis or from damaged and dying cells. Once in the extracellular environment, ATP binds to the specific receptors termed P2, which mediate ATP effects and are present broadly in both neurons and glial cells. There are P2X, the ligand-gated ionotropic receptors, possessing low affinity for ATP and responsible for fast excitatory neurotransmission, and P2Y, the metabotropic G-protein-coupled receptors, possessing high affinity for ATP. Since massive extracellular release of ATP often occurs after stress, brain ischemia and trauma, the extracellular ATP is considered relating to or involving in the pathological processes of many nervous system diseases. Conversely, the trophic functions have also been extensively described for the extracellular ATP. Therefore, extracellular ATP plays a very complex role in the CNS and its binding to P2 receptors can be related to toxic and/or beneficial effects. In this review, we described the extracellular ATP acting via P2 receptors as a potent therapeutic target for treatment of nervous system diseases. PMID- 19190687 TI - Perivascular space and neurological disorders. AB - Perivascular space (PVS) is a crevice between two slices of cerebral pia maters, filled with tissue fluid, which be formed by pia mater emboling in the surrounding of cerebral perforating branch (excluding micrangium). Normal PVS (diameter < 2 mm) can be found in almost all healthy adults; however enlarged PVS (diameter > 2 mm) has correlation with neurological disorders probably. The article reviews the formation mechanism, imageology characteristics and the relation with neurological disorders of PVS, which is beneficial to the research of some neurological disorders etiopathogenesis and treatment. PMID- 19190688 TI - Review of the diagnosis and clinical features of multiple sclerosis in China. AB - This review focused on the diagnosis and clinical features of multiple sclerosis (MS) in China. We have identified the published researching information from 1976 to 2008 in China. The key issues related to the diagnosis and clinical features of MS in China were summarized. The first patient with MS in China was reported in 1926 from Xiehe hospital. Case reports on MS have been increasing during recent decades. Almost all the patients with MS were confirmed by the McDonald criteria (1977) before 1984. After the year of 1992, even to this day, the Poser criteria were widely used in China. Although the new diagnostic criteria, McDonald criteria (2001), were presented in 2001, only few papers published in Chinese were reported. The most frequent initial symptoms or signs of the patients with MS were optic nerve, motor weakness and sensory symptoms. The most frequent location of MS lesions over the course was found to be the spinal cord, followed by the cerebrum and optic nerves. Almost all patients had been treated with corticosteroids. This review supported previous observations in Chinese patients with MS. However, further studies are needed to understand epidemiologic features of MS in China. PMID- 19190689 TI - Review article composite lymphoma. AB - Background : Composite lymphoma (CL) is a rare disease that has been identified in recent literature. The term composite lymphoma was first proposed to denote the occurrence of more than one lymphoma in a single patient; however, the present accepted definition is the occurrence of 2 or more distinct lymphoma types in a single anatomic site. The condition could be concurrent or sequential. Unlike disease progression or transformation in lymphoma, CL should include two distinct clones proven by morphological and laboratory tests. Pathogenesis : No single definite mechanism has been suggested to explain the pathogenesis of the different types of CL. The etiology is variable, complex and differs according to the types of lymphomas involved. Several theories were proposed including clonal selection with additional mutational accumulation, genomic instability with genetic predisposition, common precursor cell and the aid of a viral factor, mostly EBV. Diagnosis : The morphologic criteria must be confirmed by one or more tests including immunohistochemistry, flow cytometry, gene rearrangement by PCR, cytogenetics, FISH, in-situ hybridization, DNA sequencing and cDNA microarray . Results are more accurate using the laser capture microdissection method. Many combinations of CL are reported, including : Multiple B-cell lymphomas; B-cell and T-cell lymphomas; NHL and HL; or complex B-cell, T-cell and HL cases. Conclusion : Due to the great advancement in molecular characterization of lymphoma, CL is being increasingly identified. It must be carefully diagnosed, because the multiple disease entities may have entirely different natural histories, prognosis and treatment modalities. Also, careful study of such cases may clarify the possible pathogenic mechanisms of the interrelationship of clonal evolution in lymphoma. Key Words : Composite lymphoma -EBV. PMID- 19190690 TI - ERCC1 Expression in Diffuse Large B-Cell Lymphoma Patients Treated with a Cisplatin-Based Regimen : A Brief Communication. AB - There is growing interest in defining biomarkers that could predict response to different chemotherapeutic agents. Excision repair cross complement 1 (ERCC1) enzyme has been shown to predict benefit of cisplatin in different types of cancer. As cisplatin-based regimens are frequently used in the salvage treatment of diffuse large B-cell lymphomas (DLBCL), a small pilot study was conducted to determine whether ERCC1 is expressed in this disease or not. Out of seven patients examined, only one had a 25% ERCC1 expression, which could represent a tumour truly expressing this marker. However, expression was not able to predict response to treatment. It remains unclear whether ERCC1 could serve as a predictive marker for cisplatin in this disease requiring further studies. Key Words : ERCC1 -Large B-cell lymphoma -Cisplatin resistance. PMID- 19190691 TI - Missed breast carcinoma; why and how to avoid? AB - Introduction : Despite the advances in mammography techniques, it still has a number of limitations. It is estimated that about 10 to 25% of lesions are overlooked in mammograms out of which about two thirds are detected retrospectively by radiologists and oncologists. Causes of missed breast cancer on mammography can be secondary to many factors including those related to the patient (whether inherent or acquired), the nature of the malignant mass itself, poor mammographic techniques, provider factors or interpretive skills of radiologists and oncologists (including perception and interpretation errors). Aim of Work : The aim of this study is to investigate the aforementioned factors hindering early breast cancer detection and in turn lowering mammographic sensitivity and to outline the major guidelines to overcome these factors aiming to an optimum mammographic examination and interpretation by radiologists and oncologists. Subject and Methods : We conducted this multicenter study over a two year interval. We included 152 histopathologicaly proven breast carcinomas that were initially missed on mammography. The cases were subjected to mammography, complementary US, MRI and digital mammography in some cases and all cases were histopathologically proven either by FNAB, CNB or open biopsy. Results : Revision of the pathological specimens of these 152 cases revealed 121 infiltrating ductal carcinomas, 2 lobular, 4 mucinous, 14 inflammatory carcinomas, 6 carcinomas in situ (3 of which were intracystic), 2 intraductal papillary carcinomas and 3 cases with Paget's disease of the nipple. In analyzing the causes responsible for misdiagnosis of these carcinomas we classified them into 4 causative factors; patient, tumor, technical or provider factors. Tumor factors were the most commonly encountered, accounting for 44.1%, while provider factors were the least commonly encountered in 14.5 %. Carcinomas were detected using several individual or combined complementary techniques. These techniques mainly included double reading, additional mammography views, ultrasound and MRI examinations. Forty four carcinomas were detected on double and re-reading by more experienced radiologists. Additional mammographic views were recommended in 35 (23%) cases. Complementary ultrasound examination was performed for all 152 cases (100%) and showed a higher sensitivity than mammography in carcinoma detection. It was diagnostic in 138 (90.8%) cases only. In the remaining 14 cases, further MRI and biopsy were performed. Conclusion : Why can breast carcinoma be missed? Four main factors are responsible for missing a carcinoma: (1) Patient factors (Inherently dense breasts or acquired dense breasts). (2) Tumor factors (subtle carcinoma, masked carcinoma, multifocal carcinoma and multicentric carcinoma). (3) Technical factors (bad exposure factors, malpositioned breasts and bad processing quality). (4) Provider factors (bad perception and misinterpretation). How to avoid missing a breast carcinoma? Review clinical data and use US and other adjunct techniques as MRI and biopsy to assess a palpable or mammographically detected mass. Be strict about positioning and technical factors. Try to optimize image quality. Be alert to subtle features of breast cancers. Always consider the well defined carcinoma. Compare current images with multiple prior studies to look for subtle increases in lesion size. Look for other lesions when one abnormality is seen. Judge a lesion by its most malignant features. Double reading and the use of computer aided diagnosis (CAD) and finally FFDM (Full Field Digital Mammography). Close cooperation between the oncologist, radiologist and pathologist is essential to avoid missing any case of breast carcinoma. Key Words : Missed breast carcinoma -Mammography - Ultrasonography -MRI. PMID- 19190692 TI - Immunohistochemical Expression of CD10 in Cutaneous Basal and Squamous Cell Carcinomas. AB - Background : CD10 is a zinc-dependent metallopeptidase known as common acute lymphoblastic leukemia antigen (CALLA). Although CD10 expression has been investigated in some cutaneous tumors, to our knowledge, data regarding its expression in cutaneous epithelial neoplasms are very limited. We aimed to determine the immunohistochemical expression of CD10 in basal cell carcinoma (BCC) and squamous cell carcinoma (SCC) and to associate it with the available clinicopathological parameters in both tumors. Patients and Methods : This study included 16 SCC and 21 BCC cases (17 solid type, 2 morphea type and 2 adenoid basal types). BCC cases were divided into 12 cases with microscopic infiltrative base and 9 cases with well-circumscribed base. The localization of anti-CD10 to the tumor and/or stromal cells was determined in each case. Results : Positive CD10 staining was identified as brown cytoplasmic, with or without cell membrane staining. In all the 16 SCC cases, tumor cells failed to stain with CD10 in contrast to the stromal cells that showed CD10 expression in 13 cases (81%). In BCC cases, the expression of CD10 was noted in tumor cells in 10 cases (47.6%) and in stromal cells of 20 cases (95.24%). Most of CD10+ (7/10) BCC showed well circumscribed deep margin, however, most of CD10- cases (9/11) showed infiltrating base (p=0.030). BCCs with infiltrating deep margins (12 cases) tended to show CD10 negative basaloid cells (9/12) and CD10 positive stromal cells (12/12) (p=0.0003). Conclusion : From our results we suggest that CD10 might be a useful immunohistochemical marker to differentiate between BCC and SCC. At least, if tumor cells were CD10 positive, this would favor BCC over SCC. Absence of CD10 in all the SCC and in infiltrating BCC together with its overexpression in the surrounding stromal cells might confer invasive properties to such tumors. However, its relation to other poor prognostic factors needs larger studies to be confirmed. Key Words : CD 10 -Immunostaining -Skin. PMID- 19190693 TI - P63 and cytokeratin8/18 expression in breast, atypical ductal hyperplasia, ductal carcinoma in situ and invasive duct carcinoma. AB - Background and Purpose : The pattern and distribution of p63 expression as a myoepithelial/basal stem cell marker can be different between atypical ductal hyperplasia (ADH), ductal carcinoma in situ (DCIS) and invasive ductal carcinoma (IDC) and may denote basal phenotype of breast ductal carcinoma. CK8/18 is a luminal marker and may indicate a luminal phenotype of IDC and its expression in ADH and DCIS may refer to a possible precursor lesion to IDC. This work was designed to study and compare the expression of p63 and cytokeratin 8/18 (CK8/18) in some cases of ADH, DCIS and IDC. Materials and Methods : Histopathological evaluation and immunohistochemical study of anti-p63 and anti- CK8/18 was performed on selected archival cases of 7 ADH, 12 DCIS, 30 IDC of known clinicopathological data and previous estrogen receptor status (ER) for IDC. Confirmatory anti-smooth muscle actin (ASMA) expression for positive p63 cases was performed. Results : p63 was expressed in the peripheral rim of the myoepithelial cell layer in ADH and DCIS with occasional gabs in DCIS. It was positive and stained occasional malignant cells in 3/30 (10%) of IDC cases. Confirmatory ASMA staining decorated the same peripheral rim of cells in ADH and DCIS, but was negative in p63 positive IDC cases. CK8/18 was positive in 100% of ADH, 8/12 (66.7%) of DCIS and 22/30 (73%) of IDC cases. Combined p63 and CK8/18 expression was noticed in 3/30 (10%) of IDC. Conclusion : It is concluded from this study that p63 is specific and valuable in differentiating myoepithelial cells and is more specific and valuable than other myoepithelial markers, as ASMA and can differentiate between ADH, DCIS, IDC as it stains peripheral myoepithelial cells in ADH and DCIS with gabs in the latter and does not stain any neoplastic cells. In IDC, it is positive in malignant cells in a minority of cases which may indicate basal/stem cell/myoepithelial cell origin of breast carcinoma. Comparatively, CK8/18 cannot differentiate ADH, DCIS and IDC as there is no difference in its staining pattern among them, which may suggest that they are a continuum or that ADH and DCIS are precursors for the luminal phenotype of IDC. Key Words : p63 -CK8/18 -IDC -ADH -DCIS -Basal/ stem cells. PMID- 19190694 TI - Detection of bcl-2 Translocation in Patients with Chronic Hepatitis C and its Possible Relation to Antiviral Therapy: Preliminary Study. AB - It has been suggested that t(14;18) translocation of bcl-2 to the immunoglobulin heavy chain (IgH) locus may contribute to the pathogenesis of lymphoproliferative disorders (LPD) related to hepatitis C virus (HCV) infection. The present study aimed to assess the prevalence of bcl-2 translocation in Egyptian chronic HCV patients and to investigate the effect of combination antiviral therapy of interferon a and ribavirin on t(14;18). Fifty five chronic HCV patients were studied for the prevalence of t(14;18). These patients were classified into 2 groups, 33 non treated HCV patients and 22 treated HCV patients with antiviral therapy as well as control group of age and sex matched individuals. The bcl 2/IgH rearrangement was detected in peripheral blood mononuclear cells (PBMCs) by nested polymerase chain reaction. All patients have undergone HCV viral determination by real time PCR. Bcl-2/IgH translocation was detected in 21 (38.2%) of all 55 chronically infected HCV patients. Considering all patients with chronic HCV-infection, bcl-2 rearrangement was slightly more frequent in the non treated group than in those who underwent treatment with interferon plus ribavirin but the difference was not statistically significant, although treated patients showed biochemical and virologic response at the end of 6 months of antiviral therapy. In conclusion, t(14;18) in PBMCs is a frequent finding in chronic HCV infection. Key Words : Hepatitis C virus -Lymphoproliferative disorder -t(14;18) -Interferon. PMID- 19190695 TI - Maxillary swing approach for surgical resection of recurrent nasopharyngeal tumors. AB - Purpose : To evaluate the efficacy of salvage nasopharyngectomy for locally recurrent nasopharyngeal tumors through the maxillary swing approach. Methods : Antero -lateral access to the nasopharynx and its vicinity through the maxillary swing approach was used. Through a Weber-Fergusson-Longmire incision, the whole maxilla is freed and swung laterally while remaining attached to the masseter muscle and cheek flap. The nasopharynx, para-pharyngeal space and infratemporal fossa are now exposed facilitating complete resection of the tumor. Results : Seven patients were treated with this approach at the National Cancer Institute, Cairo University. There were 6 males and one female with a range of age 16-57 years. Three patients had recurrent carcinoma of the nasopharynx after primary radiation therapy, while four patients had locally advanced recurrent juvenile nasopharyngeal angiofibroma (JNA). Using the operating microscope, complete tumor resection was achieved in all patients except one patient with intracranial extension of JNA. There was no surgical mortality. Complications occurred in three patients, one patient developed osteoradionecrosis following re irradiation, a patient had epiphora while palatal fistula occurred in another patient. The follow-up period ranged from 3 months to 6 years, one patient died from distant disease and another patient is alive with residual intra-cranial disease. Conclusion : Salvage nasopharyngectomy is a valid option for selected patients with recurrent nasopharyngeal tumors. The maxillary swing approach allows safe and complete resection for nasopharyngeal carcinoma as well as juvenile angiofibroma with intracranial extension. Key Words : Recurrent nasopharyngeal tumors -Salvage nasopharyngectomy -Juvenile nasopharyngeal angiofibroma. PMID- 19190696 TI - Microbial genesis, life and death in glacial ice. AB - Arguments are given that terrestrial RNA and DNA may have originated in a frozen environment more than 4 billion years ago. Scenarios are developed for atmospheric transport of microbes onto glacial ice, their adaptation to subzero temperatures in the ice, and their incorporation into one of three habitats - liquid veins, mineral grain surfaces, or isolated inside 1 of the crystals that make up polycrystalline ice. The Arrhenius dependence of microbial metabolic rate on temperature is shown to match that required to repair damage owing to spontaneous DNA depurination and amino acid racemization. Even for the oldest glacial ice, microbial lifetime is shown not to be shortened by radiation damage from 238U, 232Th, or 40K in mineral dust in ice, by phage-induced lysis, or by penetrating cosmic radiation. Instead, death of those cells adapted to the hostile conditions in glacial ice is probably due to exhaustion of available nutrients. By contrast, in permafrost microbial death is more likely due to alpha particle radiation damage from U and Th in the soil and rocks intermixed with ice. For residence times in ice longer than a million years, spore formers may be unable to compete in longevity with vegetative cells that are able to repair DNA damage via survival metabolism. PMID- 19190697 TI - Culturable microbes in shallow groundwater underlying ornithogenic soil of Cape Hallett, Antarctica. AB - The objective of this study was to investigate the culturable psychrotolerant microbial community in groundwater from Seabee Hook, Antarctica. Shallow groundwater can be present in coastal regions at higher latitudes during the Antarctic summer. Perched groundwater atop ice-cemented permafrost occurs on Seabee Hook, Cape Hallett, at depths from 5 to 80 cm below the soil surface. Compared with terrestrial water from other sites in Antarctica, the groundwater was high in salt and nutrients, reflecting proximity to the sea and ornithogenic soil. Microbial communities in groundwater samples from Seabee Hook exhibited aerobic metabolism of 14C-acetate at 5 degrees C. Numbers of culturable aerobic heterotrophs in the samples ranged from <10 to ca. 1 x 106 colony-forming units.mL-1, and similar numbers of microaerophiles and nitrate reducers were detected. In contrast, numbers of nitrifiers, sulfate reducers, and iron reducers were up to 1000-fold lower. All cultures were incubated at 5 degrees C. Aerobic heterotrophic bacteria isolated from the groundwater were assigned to Actinobacteria, Proteobacteria, or Bacteroidetes. The isolates were most similar to cultured bacteria from Antarctic soil or sediment and were cold, salt, and alkaline pH tolerant, indicating they are adapted to in situ conditions. PMID- 19190698 TI - Bacterial diversity associated with ornithogenic soil of the Ross Sea region, Antarctica. AB - In the Ross Sea region of Antarctica, ornithogenic soils form on land under Adelie Penguin rookeries. Compared with mineral soils of the Ross Sea region, ornithogenic soils are generally high in microbial biomass, organic carbon, and total nitrogen and phosphorus, with high electrical conductivity and large variations in pH. The objective of this study was to assess the bacterial composition of ornithogenic soils from Cape Hallett and Cape Bird in the Ross Sea region using culture-independent methods. Soil clone libraries were constructed and those clones that occurred > or = 3 times were sequenced. The bacterial diversity of the soils was dependent on the presence of penguins. Firmicutes most closely related to the endospore-formers (e.g., Oceanobacillus profundus and Clostridium acidurici) and (or) Gammaproteobacteria belonging to the genus Psychrobacter dominated soils currently occupied with penguins. In contrast, Gammaproteobacteria, closely related to cultured members of the genera Rhodanobacter, Psychrobacter, Dokdonella, and Lysobacter, dominated the soils previously colonized by penguins. Results of this study indicate that despite relatively high nutrient levels and microbial biomass, bacterial communities of ornithogenic soils were not more diverse than those of mineral soils of the Ross Sea region of Antarctica. PMID- 19190699 TI - Antibiotic resistance among bacteria isolated from seawater and penguin fecal samples collected near Palmer Station, Antarctica. AB - Antibiotic resistance in aquatic bacteria has increased steadily as a consequence of the widespread use of antibiotics, but practice and international treaty should have limited antibiotic contamination in Antarctica. We estimated antibiotic resistance in microorganisms isolated from the Antarctic marine waters and a penguin rookery, for 2 reasons: (i) as a measure of human impact and (ii) as a potential "snapshot" of the preantibiotic world. Samples were taken at 4 established sampling sites near Palmer Station, which is situated at the southern end of the Palmer Archipelago (64 degrees 10'S, 61 degrees 50'W). Sites were chosen to provide different potentials for human contamination. Forty 50 mL samples of seawater were collected and colony-forming units (CFU)/mL were determined at 6 and 20 degrees C. For this study, presumed psychrophiles (growth at 6 degrees C) were assumed to be native to Antarctic waters, whereas presumed mesophiles (growth at 20 degrees C but not at 6 degrees C) were taken to represent introduced organisms. The 20-6 degrees C CFU/mL ratio was used as a measure of the relative impact to the ecosystem of presumably introduced organisms. This ratio was highest at the site nearest to Palmer Station and decreased with distance from it, suggesting that human presence has impacted the natural microbial flora of the site. The frequency of resistance to 5 common antibiotics was determined in each group of isolates. Overall drug resistance was higher among the presumed mesophiles than the presumed psychrophiles and increased with proximity to Palmer Station, with the presumed mesophiles showing higher frequencies of single and multiple drug resistance than the psychrophile population. The frequency of multidrug resistance followed the same pattern. It appears that multidrug resistance is low among native Antarctic bacteria but is increased by human habitation. PMID- 19190700 TI - Investigations of fungal diversity in wooden structures and soils at historic sites on the Antarctic Peninsula. AB - Investigations of microbial diversity in Antarctic are important to begin to understand ecosystem functioning and decomposition processes. This study documents fungi at 9 historic sites on the Antarctic Peninsula collected from wooden structures, other organic materials, and soils during a joint National Science Foundation and British Antarctic Survey expedition in 2007. Many of these sites had wooden structures built by the British during the World War II Operation Tabarin, but others visited included the American "East Base" on Stonington Island and the Swedish hut on Snow Hill Island. Fungi were cultured on several different media and pure cultures were obtained and identified by DNA sequencing of the internal transcribed spacer region. Cadophora species previously found to attack historic wooden structures on Ross Island, Antarctica, were found at all but 1 location sampled in the Peninsula region. Fungi causing decay in the historic wooden structures and artifacts and those causing mold problems inside the structures are of great concern, and conservation efforts are urgently needed to help preserve these important polar heritage structures. The results presented also expand our knowledge on the identity of fungi present throughout the Antarctic Peninsula region and provide insights into the organisms responsible for decomposition and nutrient recycling. PMID- 19190701 TI - The water-borne protein signals (pheromones) of the Antarctic ciliated protozoan Euplotes nobilii: structure of the gene coding for the En-6 pheromone. AB - The marine Antarctic ciliate, Euplotes nobilii, secretes a family of water-borne signal proteins, denoted as pheromones, which control vegetative proliferation and mating in the cell. Based on the knowledge of the amino acid sequences of a set of these pheromones isolated from the culture supernatant of wild-type strains, we designed probes to identify their encoding genes in the cell somatic nucleus (macronucleus). The full-length gene of the pheromone En-6 was determined and found to contain an open-reading frame specific for the synthesis of the En-6 cytoplasmic precursor (pre-pro-En-6), which requires 2 proteolytic cleavages to remove the signal peptide (pre) and the prosegment before secretion of the mature protein. In contrast to the sequence variability that distinguishes the secreted pheromones, the pre- and pro-sequences appear to be tightly conserved and useful for the construction of probes to clone every other E. nobilii pheromone gene. Potential intron sequences in the coding region of the En-6 gene imply the synthesis of more En-6 isoforms. PMID- 19190702 TI - Production of cryoprotectant extracellular polysaccharide substances (EPS) by the marine psychrophilic bacterium Colwellia psychrerythraea strain 34H under extreme conditions. AB - Extracellular polysaccharide substances (EPS) play critical roles in microbial ecology, including the colonization of extreme environments in the ocean, from sea ice to the deep sea. After first developing a sugar-free growth medium, we examined the relative effects of temperature, pressure, and salinity on EPS production (on a per cell basis) by the obligately marine and psychrophilic gamma proteobacterium, Colwellia psychrerythraea strain 34H. Over growth-permissive temperatures of approximately 10 to -4 degrees C, EPS production did not change, but from -8 to -14 degrees C when samples froze, EPS production rose dramatically. Similarly, at growth-permissive hydrostatic pressures of 1-200 atm (1 atm = 101.325 kPa) (at -1 and 8 degrees C), EPS production was unchanged, but at higher pressures of 400 and 600 atm EPS production rose markedly. In salinity tests at 10-100 parts per million (and -1 and 5 degrees C), EPS production increased at the freshest salinity tested. Extreme environmental conditions thus appear to stimulate EPS production by this strain. Furthermore, strain 34H recovered best from deep-freezing to -80 degrees C (not found for Earthly environments) if first supplemented with a preparation of its own EPS, rather than other cryoprotectants like glycerol, suggesting EPS production as both a survival strategy and source of compounds with potentially novel properties for biotechnological and other applications. PMID- 19190703 TI - Bacterial community structure and carbon turnover in permafrost-affected soils of the Lena Delta, northeastern Siberia. AB - Arctic permafrost environments store large amounts of organic carbon. As a result of global warming, intensified permafrost degradation and release of significant quantities of the currently conserved organic matter is predicted for high latitudes. To improve our understanding of the present and future carbon dynamics in climate sensitive permafrost ecosystems, the present study investigates structure and carbon turnover of the bacterial community in a permafrost-affected soil of the Lena Delta (72 degrees 22'N, 126 degrees 28'E) in northeastern Siberia. 16S rRNA gene clone libraries revealed the presence of all major soil bacterial groups and of the canditate divisions OD1 and OP11. A shift within the bacterial community was observed along the soil profile indicated by the absence of Alphaproteobacteria and Betaproteobacteria and a simultaneous increase in abundance and diversity of fermenting bacteria like Firmicutes and Actinobacteria near the permafrost table. BIOLOG EcoPlates were used to describe the spectrum of utilized carbon sources of the bacterial community in different horizons under in situ temperature conditions in the presence and absence of oxygen. The results revealed distinct qualitative differences in the substrates used and the turnover rates under oxic and anoxic conditions. It can be concluded that constantly negative redox potentials as characteristic for the near permafrost table horizons of the investigated soil did effectively shape the structure of the indigenous bacterial community limiting its phylum-level diversity and carbon turnover capacity. PMID- 19190704 TI - Mycorrhizal fungi supply nitrogen to host plants in Arctic tundra and boreal forests: 15N is the key signal. AB - Symbiotic fungi's role in providing nitrogen to host plants is well-studied in tundra at Toolik Lake, Alaska, but little-studied in the adjoining boreal forest ecosystem. Along a 570 km north-south transect from the Yukon River to the North Slope of Alaska, the 15N content was strongly reduced in ectomycorrhizal and ericoid mycorrhizal plants including Betula, Salix, Picea mariana (P. Mill.) B.S.P., Picea glauca Moench (Voss), and ericaceous plants. Compared with the 15N content of soil, the foliage of nonmycorrhizal plants (Carex and Eriophorum) was unchanged, whereas content of the ectomycorrhizal fungi was very much higher (e.g., Boletaceae, Leccinum and Cortinarius). It is hypothesized that similar processes operate in tundra and boreal forest, both nitrogen-limited ecosystems: (i) mycorrhizal fungi break down soil polymers and take up amino acids or other nitrogen compounds; (ii) mycorrhizal fungi fractionate against 15N during production of transfer compounds; (iii) host plants are accordingly depleted in 15N; and (iv) mycorrhizal fungi are enriched in 15N. Increased N availability for plant roots or decreased light availability to understory plants may have decreased N allocation to mycorrhizal partners and increased delta15N by 3-4 parts per million for southern populations of Vaccinium vitis-idaea L. and Salix. Fungal biomass, measured as ergosterol, correlated strongly with soil organic matter and attained amounts similar to those in temperate forest soils. PMID- 19190705 TI - A computer-based approach for assessing dietary supplement use in conjunction with dietary recalls. AB - In response to the need to assess both food and supplemental sources of nutrients, we have expanded the capabilities of Nutrition Data System for Research (NDSR) software to allow for assessing dietary supplement use. A Dietary Supplement Assessment Module allows for the automated collection and coding of dietary supplement use. The module is designed for use in conjunction with the software's 24-hour dietary recall features. The medication inventory method, commonly used in pharmaceutical research, served as the basis for the module's assessment approach. In adapting this approach for use in our software we designed a tiered structure that involves first screening for use of dietary supplements, then collecting product detail (e.g. full name of product, number of times taken, etc.), and finally reviewing the information with the participant. Preliminary results from a demonstration study being conducted to evaluate the Module indicate the assessment approach is acceptable to both participants and interviewers. Collecting dietary supplement use information significantly increases interview time, especially for those using multiple products. A validation study is needed to determine whether the new method results in accurate estimation of nutrient intake from supplemental sources. PMID- 19190706 TI - Harm reduction ethics: Acknowledging the values and beliefs behind our actions. PMID- 19190707 TI - Transfer of Old 'Reactivated' Memory Retrieval Cues in Rats. AB - The present studies examined whether the retrieval of an old 'reactivated' memory could be brought under the control of new contextual cues. In Experiment 1 rats trained in one context were exposed to different contextual cues either immediately, 60 min, or 120 min after a cued reactivation of the training memory. When tested in the shifted context, subjects exposed shortly after reactivation treated the shifted context as the original context. This transfer diminished with longer post-reactivation delays. Experiment 2 replicated the basic finding and demonstrated that the transfer of the old retrieval cues was specific to the contextual cues present during exposure. These findings are consistent with previous research (i.e., Briggs, Fitz, & Riccio, in press) showing the transfer of retrieval cues for a new memory, and demonstrating a similarity (in this case) between newly acquired and old reactivated memories. PMID- 19190708 TI - Development of the Iowa bone nutrient food frequency questionnaire based on data from the US Department of Agriculture Continuing Survey of the Food Intake by Individuals. AB - An easily administered food frequency questionnaire (FFQ)/dietary screener was developed for current (adult) and retrospective (adolescent) intakes of nutrients important for bone development and maintenance. This tool quantified serving sizes and nutrients from foods using gender and age specific techniques. Nutrients of interest were calcium, vitamin D, caffeine and alcohol, and 15 categories of foods were selected for inclusion based on frequency of intake and nutrient density. Calcium-contributing foods were selected from published dietary intake assessment tools. Foods contributing vitamin D, caffeine and alcohol were selected based on nutrient density and Midwest consumption practices. Serving sizes were quantified in standard serving units or as small, medium and large servings. Food items selected for the FFQ/dietary screener were matched to foods from the United States Department of Agriculture (USDA) Continuing Survey of Food Intakes by Individuals (CSFII). Calcium, caffeine and alcohol values were assigned using CSFII data files at median values per 100g intake. CSFII midpoint tertile frequency of intake values for males and females 14-18 and 25-45 years old were used to establish serving weights for small, medium and large servings. CSFII data files provide an efficient way for estimating typical intakes, serving sizes and nutrient values for target groups. Age- and gender-derived data provided realistic estimates of nutrient intakes when using FFQ/dietary screener assessment method. PMID- 19190709 TI - Differentiation Between Low Positive Affectivity and Behavioral Inhibition in Preschool-Age Children: A Comparison of Behavioral Approach in Novel and Non Novel Contexts. AB - The temperament constructs of low positive affectivity (PA) and high behavioral inhibition (BI) overlap and are often not differentiated in the research literature. In particular, both constructs are characterized by low approach and engagement. However, current theoretical conceptualizations of these constructs suggest that low PA should be associated with low approach across most contexts, whereas BI should be associated with low approach only in novel situations. The present study used laboratory measures of child temperament and behavior to test these hypotheses in a sample of 100 preschool-age children. Results indicated that in novel situations, both lower positive affect and higher BI predicted low behavioral approach. However, in non-novel situations, only lower levels of positive affect predicted lower levels of approach; BI was not related to approach behavior in more familiar contexts. In conclusion, this study indicates that the overlap between the temperament traits of low PA and high BI is limited to novel contexts and that these constructs are distinguished by behavior in non novel situations. PMID- 19190710 TI - Annealing and the Normalized N-Cut. AB - We describe an annealing procedure that computes the normalized N-cut of a weighted graph G. The first phase transition computes the solution of the approximate normalized 2-cut problem, while the low temperature solution computes the normalized N-cut. The intermediate solutions provide a sequence of refinements of the 2-cut that can be used to split the data to K clusters with 2 99:1 enantiomeric ratios. PMID- 19190742 TI - Recent progress on the total synthesis of acetogenins from Annonaceae. AB - An overview of recent progress on the total synthesis of acetogenins from Annonaceae during the past 12 years is provided. These include mono tetrahydrofurans, adjacent bis-tetrahydrofurans, nonadjacent bis tetrahydrofurans, tri-tetrahydrofurans, adjacent tetrahydrofuran tetrahydropyrans, nonadjacent tetrahydrofuran-tetrahydropyrans, mono tetrahydropyrans, and acetogenins containing only gamma-lactone. This review emphasizes only the first total synthesis of molecules of contemporary interest and syntheses that have helped to correct structures. In addition, some significant results on the novel synthesis and structure-activity relationship studies of annonaceous acetogenins are also introduced. PMID- 19190744 TI - Chemoselective reduction of aldehydes by ruthenium trichloride and resin-bound formates. AB - A simple, chemoselective transfer hydrogenation of aryl aldehydes with the aid of Amberlite((R)) resin formate (ARF), a stable H-donor, in the presence of catalytic ruthenium trichloride is described. Aromatic aldehydes and 1,2 diketones are reduced efficiently and selectively, while aryl ketones remain unchanged. Several other potentially reducible groups attached to the aromatic moiety are unaffected. PMID- 19190746 TI - Journals under threat: A joint response from history of science, technology and medicine editors. PMID- 19190747 TI - Psychobiology, psychiatry, and psychoanalysis: the intersecting careers of Adolf Meyer, Phyllis Greenacre, and Curt Richter. PMID- 19190748 TI - The politics of professionalization: MPs, Medical Men, and the 1858 Medical Act. PMID- 19190745 TI - Functional analysis of the cathepsin-like cysteine protease genes in adult Brugia malayi using RNA interference. AB - BACKGROUND: Cathepsin-like enzymes have been identified as potential targets for drug or vaccine development in many parasites, as their functions appear to be essential in a variety of important biological processes within the host, such as molting, cuticle remodeling, embryogenesis, feeding and immune evasion. Functional analysis of Caenorhabditis elegans cathepsin L (Ce-cpl-1) and cathepsin Z (Ce-cpz-1) has established that both genes are required for early embryogenesis, with Ce-cpl-1 having a role in regulating in part the processing of yolk proteins. Ce-cpz-1 also has an important role during molting. METHODS AND FINDINGS: RNA interference assays have allowed us to verify whether the functions of the orthologous filarial genes in Brugia malayi adult female worms are similar. Treatment of B. malayi adult female worms with Bm-cpl-1, Bm-cpl-5, which belong to group Ia of the filarial cpl gene family, or Bm-cpz-1 dsRNA resulted in decreased numbers of secreted microfilariae in vitro. In addition, analysis of the intrauterine progeny of the Bm-cpl-5 or Bm-cpl Pro dsRNA- and siRNA-treated worms revealed a clear disruption in the process of embryogenesis resulting in structural abnormalities in embryos and a varied differential development of embryonic stages. CONCLUSIONS: Our studies suggest that these filarial cathepsin like cysteine proteases are likely to be functional orthologs of the C. elegans genes. This functional conservation may thus allow for a more thorough investigation of their distinct functions and their development as potential drug targets. PMID- 19190749 TI - The 1815 Act To Regulate Madhouses in Scotland: a reinterpretation. PMID- 19190750 TI - "To end the degeneration of a nation": debates on eugenic sterilization in inter war Romania. PMID- 19190751 TI - Hippocratic bodily "channels" and oriental parallels. PMID- 19190752 TI - The Jernegan-Arundell correspondence. PMID- 19190753 TI - Ca(2+) permeable AMPA receptor induced long-term potentiation requires PI3/MAP kinases but not Ca/CaM-dependent kinase II. AB - Ca(2+) influx via GluR2-lacking Ca(2+)-permeable AMPA glutamate receptors (CP AMPARs) can trigger changes in synaptic efficacy in both interneurons and principle neurons, but the underlying mechanisms remain unknown. We took advantage of genetically altered mice with no or reduced GluR2, thus allowing the expression of synaptic CP-AMPARs, to investigate the molecular signaling process during CP-AMPAR-induced synaptic plasticity at CA1 synapses in the hippocampus. Utilizing electrophysiological techniques, we demonstrated that these receptors were capable of inducing numerous forms of long-term potentiation (referred to as CP-AMPAR dependent LTP) through a number of different induction protocols, including high-frequency stimulation (HFS) and theta-burst stimulation (TBS). This included a previously undemonstrated form of protein-synthesis dependent late-LTP (L-LTP) at CA1 synapses that is NMDA-receptor independent. This form of plasticity was completely blocked by the selective CP-AMPAR inhibitor IEM-1460, and found to be dependent on postsynaptic Ca(2+) ions through calcium chelator (BAPTA) studies. Surprisingly, Ca/CaM-dependent kinase II (CaMKII), the key protein kinase that is indispensable for NMDA-receptor dependent LTP at CA1 synapses appeared to be not required for the induction of CP-AMPAR dependent LTP due to the lack of effect of two separate pharmacological inhibitors (KN-62 and staurosporine) on this form of potentiation. Both KN-62 and staurosporine strongly inhibited NMDA-receptor dependent LTP in control studies. In contrast, inhibitors for PI3-kinase (LY294002 and wortmannin) or the MAPK cascade (PD98059 and U0126) significantly attenuated this CP-AMPAR-dependent LTP. Similarly, postsynaptic infusion of tetanus toxin (TeTx) light chain, an inhibitor of exocytosis, also had a significant inhibitory effect on this form of LTP. These results suggest that distinct synaptic signaling underlies GluR2-lacking CP-AMPAR dependent LTP, and reinforces the recent notions that CP-AMPARs are important facilitators of synaptic plasticity in the brain. PMID- 19190754 TI - Sequential metabolism of 7-dehydrocholesterol to steroidal 5,7-dienes in adrenal glands and its biological implication in the skin. AB - Since P450scc transforms 7-dehydrocholesterol (7DHC) to 7-dehydropregnenolone (7DHP) in vitro, we investigated sequential 7DHC metabolism by adrenal glands ex vivo. There was a rapid, time- and dose-dependent metabolism of 7DHC by adrenals from rats, pigs, rabbits and dogs with production of more polar 5,7-dienes as detected by RP-HPLC. Based on retention time (RT), UV spectra and mass spectrometry, we identified the major products common to all tested species as 7DHP, 22-hydroxy-7DHC and 20,22-dihydroxy-7DHC. The involvement of P450scc in adrenal metabolic transformation was confirmed by the inhibition of this process by DL-aminoglutethimide. The metabolism of 7DHC with subsequent production of 7DHP was stimulated by forscolin indicating involvement of cAMP dependent pathways. Additional minor products of 7DHC metabolism that were more polar than 7DHP were identified as 17-hydroxy-7DHP (in pig adrenals but not those of rats) and as pregna-4,7-diene-3,20-dione (7-dehydroprogesterone). Both products represented the major identifiable products of 7DHP metabolism in adrenal glands. Studies with purified enzymes show that StAR protein likely transports 7DHC to the inner mitochondrial membrane, that 7DHC can compete effectively with cholesterol for the substrate binding site on P450scc and that the catalytic efficiency of 3betaHSD for 7DHP (V(m)/K(m)) is 40% of that for pregnenolone. Skin mitochondria are capable of transforming 7DHC to 7DHP and the 7DHP is metabolized further by skin extracts. Finally, 7DHP, its photoderivative 20 oxopregnacalciferol, and pregnenolone exhibited biological activity in skin cells including inhibition of proliferation of epidermal keratinocytes and melanocytes, and melanoma cells. These findings define a novel steroidogenic pathway: 7DHC- >22(OH)7DHC-->20,22(OH)(2)7DHC-->7DHP, with potential further metabolism of 7DHP mediated by 3betaHSD or CYP17, depending on mammalian species. The 5-7 dienal intermediates of the pathway can be a source of biologically active vitamin D3 derivatives after delivery to or production in the skin, an organ intermittently exposed to solar radiation. PMID- 19190755 TI - Liver cancer-derived hepatitis C virus core proteins shift TGF-beta responses from tumor suppression to epithelial-mesenchymal transition. AB - BACKGROUND: Chronic hepatitis C virus (HCV) infection and associated liver cirrhosis represent a major risk factor for hepatocellular carcinoma (HCC) development. TGF-beta is an important driver of liver fibrogenesis and cancer; however, its actual impact in human cancer progression is still poorly known. The aim of this study was to investigate the role of HCC-derived HCV core natural variants on cancer progression through their impact on TGF-beta signaling. PRINCIPAL FINDINGS: We provide evidence that HCC-derived core protein expression in primary human or mouse hepatocyte alleviates TGF-beta responses in terms or growth inhibition or apoptosis. Instead, in these hepatocytes TGF-beta was still able to induce an epithelial to mesenchymal transition (EMT), a process that contributes to the promotion of cell invasion and metastasis. Moreover, we demonstrate that different thresholds of Smad3 activation dictate the TGF-beta responses in hepatic cells and that HCV core protein, by decreasing Smad3 activation, may switch TGF-beta growth inhibitory effects to tumor promoting responses. CONCLUSION/SIGNIFICANCE: Our data illustrate the capacity of hepatocytes to develop EMT and plasticity under TGF-beta, emphasize the role of HCV core protein in the dynamic of these effects and provide evidence for a paradigm whereby a viral protein implicated in oncogenesis is capable to shift TGF-beta responses from cytostatic effects to EMT development. PMID- 19190756 TI - The tree versus the forest: the fungal tree of life and the topological diversity within the yeast phylome. AB - A recurrent topic in phylogenomics is the combination of various sequence alignments to reconstruct a tree that describes the evolutionary relationships within a group of species. However, such approach has been criticized for not being able to properly represent the topological diversity found among gene trees. To evaluate the representativeness of species trees based on concatenated alignments, we reconstruct several fungal species trees and compare them with the complete collection of phylogenies of genes encoded in the Saccharomyces cerevisiae genome. We found that, despite high levels of among-gene topological variation, the species trees do represent widely supported phylogenetic relationships. Most topological discrepancies between gene and species trees are concentrated in certain conflicting nodes. We propose to map such information on the species tree so that it accounts for the levels of congruence across the genome. We identified the lack of sufficient accuracy of current alignment and phylogenetic methods as an important source for the topological diversity encountered among gene trees. Finally, we discuss the implications of the high levels of topological variation for phylogeny-based orthology prediction strategies. PMID- 19190758 TI - Adult and embryonic GAD transcripts are spatiotemporally regulated during postnatal development in the rat brain. AB - BACKGROUND: GABA (gamma-aminobutyric acid), the main inhibitory neurotransmitter in the brain, is synthesized by glutamic acid decarboxylase (GAD). GAD exists in two adult isoforms, GAD65 and GAD67. During embryonic brain development at least two additional transcripts exist, I-80 and I-86, which are distinguished by insertions of 80 or 86 bp into GAD67 mRNA, respectively. Though it was described that embryonic GAD67 transcripts are not detectable during adulthood there are evidences suggesting re-expression under certain pathological conditions in the adult brain. In the present study we systematically analyzed for the first time the spatiotemporal distribution of different GADs with emphasis on embryonic GAD67 mRNAs in the postnatal brain using highly sensitive methods. METHODOLOGY/PRINCIPAL FINDINGS: QPCR was used to precisely investigate the postnatal expression level of GAD related mRNAs in cortex, hippocampus, cerebellum, and olfactory bulb of rats from P1 throughout adulthood. Within the first three postnatal weeks the expression of both GAD65 and GAD67 mRNAs reached adult levels in hippocampus, cortex, and cerebellum. The olfactory bulb showed by far the highest expression of GAD65 as well as GAD67 transcripts. Embryonic GAD67 splice variants were still detectable at birth. They continuously declined to barely detectable levels during postnatal development in all investigated regions with exception of a comparatively high expression in the olfactory bulb. Radioactive in situ hybridizations confirmed the occurrence of embryonic GAD67 transcripts in the olfactory bulb and furthermore detected their localization mainly in the subventricular zone and the rostral migratory stream. CONCLUSIONS/SIGNIFICANCE: Embryonic GAD67 transcripts can hardly be detected in the adult brain, except for specific regions associated with neurogenesis and high synaptic plasticity. Therefore a functional role in processes like proliferation, migration or synaptogenesis is suggested. PMID- 19190757 TI - A late role for bmp2b in the morphogenesis of semicircular canal ducts in the zebrafish inner ear. AB - BACKGROUND: The Bone Morphogenetic Protein (BMP) genes bmp2 and bmp4 are expressed in highly conserved patterns in the developing vertebrate inner ear. It has, however, proved difficult to elucidate the function of BMPs during ear development as mutations in these genes cause early embryonic lethality. Previous studies using conditional approaches in mouse and chicken have shown that Bmp4 has a role in semicircular canal and crista development, but there is currently no direct evidence for the role of Bmp2 in the developing inner ear. METHODOLOGY/PRINCIPAL FINDINGS: We have used an RNA rescue strategy to test the role of bmp2b in the zebrafish inner ear directly. Injection of bmp2b or smad5 mRNA into homozygous mutant swirl (bmp2b(-/-)) embryos rescues the early patterning defects in these mutants and the fish survive to adulthood. As injected RNA will only last, at most, for the first few days of embryogenesis, all later development occurs in the absence of bmp2b function. Although rescued swirl adult fish are viable, they have balance defects suggestive of vestibular dysfunction. Analysis of the inner ears of these fish reveals a total absence of semicircular canal ducts, structures involved in the detection of angular motion. All other regions of the ear, including the ampullae and cristae, are present and appear normal. Early stages of otic development in rescued swirl embryos are also normal. CONCLUSIONS/SIGNIFICANCE: Our findings demonstrate a critical late role for bmp2b in the morphogenesis of semicircular canals in the zebrafish inner ear. This is the first demonstration of a developmental role for any gene during post embryonic stages of otic morphogenesis in the zebrafish. Despite differences in the early stages of semicircular canal formation between zebrafish and amniotes, the role of Bmp2 in semicircular canal duct outgrowth is likely to be conserved between different vertebrate species. PMID- 19190759 TI - ACC2 is expressed at high levels in human white adipose and has an isoform with a novel N-terminus [corrected]. AB - Acetyl-CoA carboxylases ACC1 and ACC2 catalyze the carboxylation of acetyl-CoA to malonyl-CoA, regulating fatty-acid synthesis and oxidation, and are potential targets for treatment of metabolic syndrome. Expression of ACC1 in rodent lipogenic tissues and ACC2 in rodent oxidative tissues, coupled with the predicted localization of ACC2 to the mitochondrial membrane, have suggested separate functional roles for ACC1 in lipogenesis and ACC2 in fatty acid oxidation. We find, however, that human adipose tissue, unlike rodent adipose, expresses more ACC2 mRNA relative to the oxidative tissues muscle and heart. Human adipose, along with human liver, expresses more ACC2 than ACC1. Using RT PCR, real-time PCR, and immunoprecipitation we report a novel isoform of ACC2 (ACC2.v2) that is expressed at significant levels in human adipose. The protein generated by this isoform has enzymatic activity, is endogenously expressed in adipose, and lacks the N-terminal sequence. Both ACC2 isoforms are capable of de novo lipogenesis, suggesting that ACC2, in addition to ACC1, may play a role in lipogenesis. The results demonstrate a significant difference in ACC expression between human and rodents, which may introduce difficulties for the use of rodent models for development of ACC inhibitors. PMID- 19190760 TI - Neuronal shot noise and Brownian 1/f2 behavior in the local field potential. AB - We demonstrate that human electrophysiological recordings of the local field potential (LFP) from intracranial electrodes, acquired from a variety of cerebral regions, show a ubiquitous 1/f(2) scaling within the power spectrum. We develop a quantitative model that treats the generation of these fields in an analogous way to that of electronic shot noise, and use this model to specifically address the cause of this 1/f(2) Brownian noise. The model gives way to two analytically tractable solutions, both displaying Brownian noise: 1) uncorrelated cells that display sharp initial activity, whose extracellular fields slowly decay in time and 2) rapidly firing, temporally correlated cells that generate UP-DOWN states. PMID- 19190761 TI - Mesoporous carbons with self-assembled surfaces of defined crystal orientation. AB - The design of carbon sorbents traditionally focuses on the control of pore structure and the number and type of surface functional groups. The present paper explores the potential of also controlling the carbon crystal structure, or graphene layer orientation, in the immediate vicinity of the internal surfaces. We hypothesize that this crystal structure influences the properties of the carbon surfaces and affects the number and type of active sites for functionalization. Here a series of mesoporous carbons are fabricated by capillary infiltration of mesophase pitch (naphthalene homopolymer) into a series of controlled pore glass templates of different characteristic pore size followed by carbonization and template etching. The liquid crystalline mesogens are known to adopt perpendicular alignment (anchoring) at liquid/silica interfaces, which after carbonization lead to a high concentration of graphene edge sites at the inner surfaces. These surfaces are shown to have elevated chemical reactivity, and the pore structures are shown to be consistent with predictions of a quantitative model based on the negative replica concept. Overall, the use of mesophase pitch for templated mesoporous carbons allows systematic and simultaneous control of both pore structure and interfacial crystal structure through the well-defined rules of liquid crystal surface anchoring. PMID- 19190762 TI - Circulating tissue inhibitor of matrix metalloproteinase-4 (TIMP-4) in systemic sclerosis patients with elevated pulmonary arterial pressure. AB - Decreased levels of matrix metalloproteinases (MMPs) or excess levels of their tissue inhibitors (TIMPs) may contribute to dysregulation of extracellular matrix turnover in systemic sclerosis (SSc). In a cross-sectional study of 106 SSc patients, we measured serum levels of TIMP-4 which is preferentially expressed in cardiovascular structures and searched for correlations with simultaneously performed echocardiography measurements of pulmonary artery systolic pressure (PASP), myocardial performance, and pulmonary function tests. TIMP-4, but not MMP 9, levels were significantly raised in patients with SSc than controls. However, in the subgroup of patients with PASP measurements lower to 40 mmHg (n = 69), TIMP-4 levels were comparable to controls irrespective of the presence of diffuse or limited skin involvement, or lung fibrosis. Individual PASP measurements suggestive of pulmonary hypertension were associated with increased TIMP-4 serum levels (P = .03), independently of age, extent of skin sclerosis, or lung fibrosis, suggesting a cardiopulmonary vasculature-specific role of TIMP-4 activation in SSc. PMID- 19190763 TI - Disruption of Nrf2 enhances upregulation of nuclear factor-kappaB activity, proinflammatory cytokines, and intercellular adhesion molecule-1 in the brain after traumatic brain injury. AB - Inflammatory response plays an important role in the pathogenesis of secondary brain injury after traumatic brain injury (TBI). Nuclear factor erythroid 2 related factor 2 (Nrf2) is a key transcription factor that plays a crucial role in cytoprotection against inflammation. The present study investigated the role of Nrf2 in the cerebral upregulation of NF-kappaB activity, proinflammatory cytokine, and ICAM-1 after TBI. Wild-type Nrf2 (+/+) and Nrf2 (-/-)-deficient mice were subjected to a moderately severe weight-drop impact head injury. Electrophoretic mobility shift assays (EMSAs) were performed to analyze the activation of nuclear factor kappa B (NF-kappaB). Enzyme-linked immunosorbent assays were performed to quantify the production of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), and interleukin-6 (IL-6). Immunohistochemistry staining experiments were performed to detect the expression of intercellular adhesion molecule-1 (ICAM-1). Nrf2 (-/-) mice were shown to have more NF-kappaB activation, inflammatory cytokines TNF-alpha, IL-1beta and IL-6 production, and ICAM-1 expression in brain after TBI compared with their wild type Nrf2 (+/+) counterparts. The results suggest that Nrf2 plays an important protective role in limiting the cerebral upregulation of NF-kappaB activity, proinflammatory cytokine, and ICAM-1 after TBI. PMID- 19190764 TI - Chronic brucellosis patients retain low frequency of CD4+ T-lymphocytes expressing CD25 and CD28 after Escherichia coli LPS stimulation of PHA-cultured PBMCs. AB - Chronic brucellosis patients display a defective Th1 response to PHA. We have previously shown that heat-killed B. abortus (HKBA) can downregulate the PHA induced increase of CD4+/CD25+ and CD14+/CD80+ cells of brucellosis patients. In the present study, we investigate the effect of E. coli LPS, as a potent stimulant of monocytes and autologous T-lymphocytes, on the PHA-cultured PBMCs of the same groups of patients. Thirteen acute brucellosis (AB) patients, 22 chronic brucellosis (CB) patients, 11 "cured" subjects, and 15 healthy volunteers were studied. The percentage of CD4+/CD25+ and CD4+/CD28+ T-lymphocytes as well as CD14+/CD80+ monocytes were analyzed by flow cytometry after PBMCs culture with PHA plus E. coli LPS. A significant decrease in the percentage of CD4+/CD25+ and CD4+/CD28+ T-lymphocytes was observed in CB compared to AB. In HKBA cultures, compared to E. coli LPS-cultures, there was a significant reduction of CD4+/CD25+ T-lymphocytes in all groups and CD14+/CD80+ in patients groups. We suggest that Brucella can modulate host immune response, leading to T-cell anergy and chronic infection. PMID- 19190765 TI - Retroperitoneal lymph nodes in transitional cell carcinoma of the kidney and ureter. AB - The incidence of transitional cell carcinoma of the kidney and ureter is low and for that reason limited data exists regarding the appropriate management of regional retroperitoneal lymph nodes. Lymph node metastases have consistently been associated with an adverse prognosis. However, five-year cancer-specific survival following nephroureterectomy and lymphadenectomy for patients with lymph node involvement ranges from 0-39%, suggesting a therapeutic benefit. This review covers the primary tumor characteristics associated with lymph node involvement, imaging of the lymph nodes, as well as the rationale, role, patient selection, suggested anatomic templates, and technical considerations for lymphadenectomy. PMID- 19190766 TI - The role of chemotherapy in upper tract urothelial carcinoma. AB - Locally advanced upper tract urothelial carcinoma has a poor prognosis. While surgery represents the only potentially curable therapeutic intervention, recurrences are common and typically systemic in nature. It is thus reasonable to consider perioperative chemotherapy in an effort to decrease the risk of recurrence. There are very little direct data providing clinical guidance in this scenario. For urothelial cancer of the bladder, there are randomized phase III data demonstrating a survival advantage with neoadjuvant cisplatin-based combination chemotherapy. Although arguments favoring adjuvant chemotherapy could be made for upper tract urothelial cancer, the loss of renal function that occurs with nephrectomy can complicate administration of appropriate perioperative treatment. Therefore, by analogy to urothelial carcinoma of the lower tract, it is argued that cisplatin-based neoadjuvant chemotherapy should be the standard of care for patients with locally advanced upper tract urothelial cancer. PMID- 19190767 TI - Evaluation of a telemedicine system for the transmission of morpho/immunological data aiming at the inclusion of patients in a therapeutic trial. AB - Due to their high levels of achievement and efficiency, image digitalization and teletransmission tools are more and more frequently used. Applied to cellular haematology, these tools often contribute to diagnosis confrontation, sometimes within the framework of therapeutic trials. We present one of the first approaches of the use of telehaematology for the inclusion of patients in the GOELAMS chronic lymphocytic leukaemia 98 trial. The advantages were (1) the creation of a unique, protected, stable data bank that could be remotely consulted, (2) the use of digitized pictures which made expertise on identical documents possible, (3) the facility of computer exchanges between experts, in terms of reception as well as replying time delays. We were able to set out new standards of image sampling for CLL, solve the semantic divergences, and point out interobserver variability as regards morphology. The limiting factors were the important need for expert investment, but they more importantly concerned the first line morphologists who should benefit from adequate tools, in terms of computer equipment as well as members of staff, so as to apprehend this second reading system as a quality control procedure. PMID- 19190768 TI - Antitumor activity of 6-(cyclohexylamino)-1, 3-dimethyl-5(2-pyridyl)furo[2,3 d]pyrimidine-2,4(1H,3H)-dione and Its Ti(IV), Zn(II), Fe(III), and Pd(II) complexes on K562 and Jurkat cell lines. AB - (6-(cyclohexylamino)-1,3-dimethyl-5(2-pyridyl)furo[2,3-d]pyrimidine-2,4(1H,3H) dione) abbreviated as CDP was synthesized and characterized. Ti(IV), Zn(II), Fe(III), and Pd(II) metal complexes of this ligand are prepared by the reaction of salts of Ti(IV), Zn(II), Fe(III), and Pd(II) with CDP in acetonitrile. Characterization of the ligand and its complexes was made by microanalyses, FT IR, (1)H NMR, (13)C NMR, and UV-Visible spectroscopy. All complexes were characterized by several techniques using elemental analysis (C, H, N), FT-IR, electronic spectra, and molar conductance measurements. The elemental analysis data suggest the stoichiometry to be 1:1 [M:L] ratio formation. The molar conductance measurements reveal the presence of 1:1 electrolytic nature complexes. These new complexes showed excellent antitumor activity against two kinds of cancer cells that are K562 (human chronic myeloid leukemia) cells and Jurkat (human T lymphocyte carcinoma) cells. PMID- 19190770 TI - An adaptive source-channel coding with feedback for progressive transmission of medical images. AB - A novel adaptive source-channel coding with feedback for progressive transmission of medical images is proposed here. In the source coding part, the transmission starts from the region of interest (RoI). The parity length in the channel code varies with respect to both the proximity of the image subblock to the RoI and the channel noise, which is iteratively estimated in the receiver. The overall transmitted data can be controlled by the user (clinician). In the case of medical data transmission, it is vital to keep the distortion level under control as in most of the cases certain clinically important regions have to be transmitted without any visible error. The proposed system significantly reduces the transmission time and error. Moreover, the system is very user friendly since the selection of the RoI, its size, overall code rate, and a number of test features such as noise level can be set by the users in both ends. A MATLAB-based TCP/IP connection has been established to demonstrate the proposed interactive and adaptive progressive transmission system. The proposed system is simulated for both binary symmetric channel (BSC) and Rayleigh channel. The experimental results verify the effectiveness of the design. PMID- 19190769 TI - Physiological role of plasmacytoid dendritic cells and their potential use in cancer immunity. AB - Dendritic cells (DCs) play a pivotal role in the control of innate and adaptive immune responses. They are a heterogeneous cell population, where plasmacytoid dendritic cells (pDCs) are a unique subset capable of secreting high levels of type I IFNs. It has been demonstrated that pDCs can coordinate events during the course of viral infection, atopy, autoimmune diseases, and cancer. Therefore, pDC, as a main source of type I IFN, is an attractive target for therapeutic manipulations of the immune system to elicit a powerful immune response against tumor antigens in combination with other therapies. The therapeutic vaccination with antigen-pulsed DCs has shown a limited efficacy to generate an effective long-lasting immune response against tumor cells. A rational manipulation and design of vaccines which could include DC subsets outside "Langerhans cell paradigm" might allow us to improve the therapeutic approaches for cancer patients. PMID- 19190771 TI - Invasive snails and an emerging infectious disease: results from the first national survey on Angiostrongylus cantonensis in China. AB - BACKGROUND: Eosinophilic meningitis (angiostrongyliasis) caused by Angiostrongylus cantonensis is emerging in mainland China. However, the distribution of A. cantonensis and its intermediate host snails, and the role of two invasive snail species in the emergence of angiostrongyliasis, are not well understood. METHODOLOGY/PRINCIPAL FINDINGS: A national survey pertaining to A. cantonensis was carried out using a grid sampling approach (spatial resolution: 40x40 km). One village per grid cell was randomly selected from a 5% random sample of grid cells located in areas where the presence of the intermediate host snail Pomacea canaliculata had been predicted based on a degree-day model. Potential intermediate hosts of A. cantonensis were collected in the field, restaurants, markets and snail farms, and examined for infection. The infection prevalence among intermediate host snails was estimated, and the prevalence of A. cantonensis within P. canaliculata was displayed on a map, and predicted for non sampled locations. It was confirmed that P. canaliculata and Achatina fulica were the predominant intermediate hosts of A. cantonensis in China, and these snails were found to be well established in 11 and six provinces, respectively. Infected snails of either species were found in seven provinces, closely matching the endemic area of A. cantonensis. Infected snails were also found in markets and restaurants. Two clusters of A. cantonensis-infected P. canaliculata were predicted in Fujian and Guangxi provinces. CONCLUSIONS/SIGNIFICANCE: The first national survey in China revealed a wide distribution of A. cantonensis and two invasive snail species, indicating that a considerable number of people are at risk of angiostrongyliasis. Health education, rigorous food inspection and surveillance are all needed to prevent recurrent angiostrongyliasis outbreaks. PMID- 19190772 TI - Association of the gene polymorphisms IFN-gamma +874, IL-13 -1055 and IL-4 -590 with patterns of reinfection with Schistosoma mansoni. AB - BACKGROUND: The immunologic findings that most consistently correlate with resistance in human schistosomiasis are high levels of IgE and low levels of IgG4. We have genotyped gene and promoter polymorphisms of cytokines associated with regulation of these isotypes in a cohort of men occupationally exposed to Schistosoma mansoni in western Kenya and evaluated their patterns with respect to resistance and susceptibility to reinfection after treatment and cure with praziquantel (PZQ). METHODOLOGY/PRINCIPAL FINDINGS: In this cohort, polymorphisms in IL-4 (-590T high IgE), IL-13 (-1055T high producer) and IFN-gamma (+874A high producer) demonstrated several correlations with resistance to reinfection. Resistance to reinfection was significantly correlated with the heterozygous IL-4 -590 genotype C/T (OR 3.5, [CI 1.2, 10.2]) compared to T/T. Among men with a homozygous IL-13 genotype CC/TT, having a T allele at the IFN-gamma +874 position increased the odds of resistance relative to individuals with the IFN-gamma +874 A/A genotype (OR = 17.5 [CI 3.0, 101.5]). Among men with homozygous A/A IFN-gamma genotype, the heterozygous IL-13 genotype C/T was associated with resistance relative to the homozygous C/C or T/T genotypes (OR = 22.5 [CI 3.5, 144.4]). No increases in odds of resistance were found in relation to the IL-13 genotype among those with a T allele in the IFN-gamma gene or in relation to the IFN-gamma genotype among those with a heterozygous IL-13 genotype. Calculation of the attributable proportion of resistance showed a significant synergistic interaction between IL-13 -1055 C/T and IL-4 -590 C/T. CONCLUSIONS: The identified polymorphisms do not by themselves confer resistance or susceptibility, but we propose that these genotypes allow the resistant phenotype to be developed and expressed upon suitable immune exposure. Based on the literature, these polymorphisms contribute to the regulation of their respective cytokines, likely leading to downstream differences in the production and interrelationships of critical defense mechanisms. PMID- 19190773 TI - Identification of lympho-epithelial Kazal-type inhibitor 2 in human skin as a kallikrein-related peptidase 5-specific protease inhibitor. AB - Kallikreins-related peptidases (KLKs) are serine proteases and have been implicated in the desquamation process of the skin. Their activity is tightly controlled by epidermal protease inhibitors like the lympho-epithelial Kazal-type inhibitor (LEKTI). Defects of the LEKTI-encoding gene serine protease inhibitor Kazal type (Spink)5 lead to the absence of LEKTI and result in the genodermatose Netherton syndrome, which mimics the common skin disease atopic dermatitis. Since many KLKs are expressed in human skin with KLK5 being considered as one of the most important KLKs in skin desquamation, we proposed that more inhibitors are present in human skin. Herein, we purified from human stratum corneum by HPLC techniques a new KLK5-inhibiting peptide encoded by a member of the Spink family, designated as Spink9 located on chromosome 5p33.1. This peptide is highly homologous to LEKTI and was termed LEKTI-2. Recombinant LEKTI-2 inhibited KLK5 but not KLK7, 14 or other serine proteases tested including trypsin, plasmin and thrombin. Spink9 mRNA expression was detected in human skin samples and in cultured keratinocytes. LEKTI-2 immune-expression was focally localized at the stratum granulosum and stratum corneum at palmar and plantar sites in close localization to KLK5. At sites of plantar hyperkeratosis, LEKTI-2 expression was increased. We suggest that LEKTI-2 contributes to the regulation of the desquamation process in human skin by specifically inhibiting KLK5. PMID- 19190774 TI - Transmission of MDR and XDR tuberculosis in Shanghai, China. AB - BACKGROUND: Multidrug-resistant (MDR) and extensively drug-resistant (XDR) tuberculosis (TB) are global health problems. We sought to determine the characteristics, prevalence, and relative frequency of transmission of MDR and XDR TB in Shanghai, one of the largest cities in Asia. METHODS: TB is diagnosed in district TB hospitals in Shanghai, China. Drug susceptibility testing for first-line drugs was performed for all culture positive TB cases, and tests for second-line drugs were performed for MDR cases. VNTR-7 and VNTR-16 were used to genotype the strains, and prior treatment history and treatment outcomes were determined for each patient. RESULTS: There were 4,379 culture positive TB cases diagnosed with drug susceptibility test results available during March 2004 through November 2007. 247 (5.6%) were infected with a MDR strain of M. tuberculosis and 11 (6.3%) of the 175 MDR patients whose isolate was tested for susceptibility to second-line drugs, were XDR. More than half of the patients with MDR and XDR were newly diagnosed and had no prior history of TB treatment. Nearly 57% of the patients with MDR were successfully treated. DISCUSSION: Transmission of MDR and XDR strains is a serious problem in Shanghai. While a history of prior anti-TB treatment indicates which individuals may have acquired MDR or XDR TB, it does not accurately predict which TB patients have disease caused by transmission of MDR and XDR strains. Therefore, universal drug susceptibility testing is recommended for new and retreatment TB cases. PMID- 19190775 TI - Using sequence similarity networks for visualization of relationships across diverse protein superfamilies. AB - The dramatic increase in heterogeneous types of biological data--in particular, the abundance of new protein sequences--requires fast and user-friendly methods for organizing this information in a way that enables functional inference. The most widely used strategy to link sequence or structure to function, homology based function prediction, relies on the fundamental assumption that sequence or structural similarity implies functional similarity. New tools that extend this approach are still urgently needed to associate sequence data with biological information in ways that accommodate the real complexity of the problem, while being accessible to experimental as well as computational biologists. To address this, we have examined the application of sequence similarity networks for visualizing functional trends across protein superfamilies from the context of sequence similarity. Using three large groups of homologous proteins of varying types of structural and functional diversity--GPCRs and kinases from humans, and the crotonase superfamily of enzymes--we show that overlaying networks with orthogonal information is a powerful approach for observing functional themes and revealing outliers. In comparison to other primary methods, networks provide both a good representation of group-wise sequence similarity relationships and a strong visual and quantitative correlation with phylogenetic trees, while enabling analysis and visualization of much larger sets of sequences than trees or multiple sequence alignments can easily accommodate. We also define important limitations and caveats in the application of these networks. As a broadly accessible and effective tool for the exploration of protein superfamilies, sequence similarity networks show great potential for generating testable hypotheses about protein structure-function relationships. PMID- 19190777 TI - Assessing the exceptionality of coloured motifs in networks. AB - Various methods have been recently employed to characterise the structure of biological networks. In particular, the concept of network motif and the related one of coloured motif have proven useful to model the notion of a functional/evolutionary building block. However, algorithms that enumerate all the motifs of a network may produce a very large output, and methods to decide which motifs should be selected for downstream analysis are needed. A widely used method is to assess if the motif is exceptional, that is, over- or under represented with respect to a null hypothesis. Much effort has been put in the last thirty years to derive P-values for the frequencies of topological motifs, that is, fixed subgraphs. They rely either on (compound) Poisson and Gaussian approximations for the motif count distribution in Erdos-Renyi random graphs or on simulations in other models. We focus on a different definition of graph motifs that corresponds to coloured motifs. A coloured motif is a connected subgraph with fixed vertex colours but unspecified topology. Our work is the first analytical attempt to assess the exceptionality of coloured motifs in networks without any simulation. We first establish analytical formulae for the mean and the variance of the count of a coloured motif in an Erdos-Renyi random graph model. Using simulations under this model, we further show that a Polya Aeppli distribution better approximates the distribution of the motif count compared to Gaussian or Poisson distributions. The Polya-Aeppli distribution, and more generally the compound Poisson distributions, are indeed well designed to model counts of clumping events. Altogether, these results enable to derive a P value for a coloured motif, without spending time on simulations. PMID- 19190776 TI - Object-place recognition learning triggers rapid induction of plasticity-related immediate early genes and synaptic proteins in the rat dentate gyrus. AB - Long-term recognition memory requires protein synthesis, but little is known about the coordinate regulation of specific genes. Here, we examined expression of the plasticity-associated immediate early genes (Arc, Zif268, and Narp) in the dentate gyrus following long-term object-place recognition learning in rats. RT PCR analysis from dentate gyrus tissue collected shortly after training did not reveal learning-specific changes in Arc mRNA expression. In situ hybridization and immunohistochemistry were therefore used to assess possible sparse effects on gene expression. Learning about objects increased the density of granule cells expressing Arc, and to a lesser extent Narp, specifically in the dorsal blade of the dentate gyrus, while Zif268 expression was elevated across both blades. Thus, object-place recognition triggers rapid, blade-specific upregulation of plasticity-associated immediate early genes. Furthermore, Western blot analysis of dentate gyrus homogenates demonstrated concomitant upregulation of three postsynaptic density proteins (Arc, PSD-95, and alpha-CaMKII) with key roles in long-term synaptic plasticity and long-term memory. PMID- 19190778 TI - Adhesion of human probiotic Lactobacillus rhamnosus to cervical and vaginal cells and interaction with vaginosis-associated pathogens. AB - OBJECTIVES: The ability of a probiotic Lactobacillus rhamnosus strain (Lcr35) to adhere to cervical and vaginal cells and to affect the viability of two main vaginosis-associated pathogens, Prevotella bivia, Gardnerella vaginalis, as well as Candida albicans was investigated. METHODS: Adhesion ability was determined in vitro with immortalized epithelial cells from the endocervix, ectocervix, and vagina. Coculture experiments were performed to count viable pathogens cells in the presence of Lcr35. RESULTS: Lcr35 was able to specifically and rapidly adhere to the three cell lines. In coculture assays, a decrease in pathogen cell division rate was observed as from 4 hours of incubation and bactericidal activity after a longer period of incubation, mostly with P. bivia. CONCLUSION: The ability of Lcr35 to adhere to cervicovaginal cells and its antagonist activities against vaginosis-associated pathogens suggest that this probiotic strain is a promising candidate for use in therapy. PMID- 19190779 TI - Infection with hepatitis C virus among HIV-infected pregnant women in Thailand. AB - OBJECTIVE: The purpose of this study was to describe the epidemiology of coinfection with hepatitis C virus (HCV) and HIV among a cohort of pregnant Thai women. METHODS: Samples from 1771 pregnant women enrolled in three vertical transmission of HIV studies in Bangkok, Thailand, were tested for HCV. RESULTS: Among HIV-infected pregnant women, HCV seroprevelance was 3.8% and the active HCV infection rate was 3.0%. Among HIV-uninfected pregnant women, 0.3% were HCV infected. Intravenous drug use by the woman was the factor most strongly associated with HCV seropositivity. Among 48 infants tested for HCV who were born to HIV/HCV coinfected women, two infants were HCV infected for an HCV transmission rate of 4.2% (95% 0.51-14.25%). CONCLUSIONS: HCV seroprevalence and perinatal transmission rates were low among this Thai cohort of HIV-infected pregnant women. PMID- 19190780 TI - The Role of PPARgamma Receptors and Leukotriene B(4) Receptors in Mediating the Effects of LY293111 in Pancreatic Cancer. AB - Pancreatic cancer is a devastating disease in which current therapies are inadequate. Separate lines of research have identified the 5 lipoxygenase/leukotriene B(4) receptor pathway and the PPARgamma pathway as potential targets for prevention or treatment of this disease. LY293111 was originally designed as a potent leukotriene B(4) receptor antagonist for treatment of inflammatory conditions. LY293111 was also known to have inhibitory effects on 5-lipoxygenase, which is upstream of the production of leukotrienes. LY293111 was shown to have potent anticancer effects in pancreatic cancer and several other solid malignancies, where it caused cell cycle arrest and marked apoptosis. Subsequently, it came to light that LY293111 exhibited PPARgamma agonist activity in addition to its effects on the 5-lipoxygenase pathway. This raises the question of which of the two targets is of greatest importance with regard to the anticancer effects of this agent. The evidence to date is not conclusive, but suggests that the effects of LY293111 may be mediated by both LTB(4) receptors and PPARgamma. PMID- 19190782 TI - KIF14 and E2F3 mRNA expression in human retinoblastoma and its phenotype association. AB - PURPOSE: We quantified mRNA expression of candidate genes for proliferation (KIF14 and E2F3) in a large retinoblastoma tumor cohort and associated with disease phenotype. METHODS: KIF14 and E2F3 mRNA expression was quantified by real time PCR in 57 retinoblastoma (RB) tumors, 3 RB cell lines, and control samples that included 4 each fetal, age-matched, adult retinas. Immunohistochemistry was done to confirm KIF14 and E2F3 protein expression in tumor cells. The mRNA expression levels were correlated with disease phenotypes including the significance of chemotherapy on tumors. RESULTS: There was statistically significant overexpression of KIF14 and E2F3 mRNA in tumors compared with control retinas (p<0.0001). Further, E2F3 also showed a significant overexpression compared to RB cell lines (p=0.01). Immunohistochemistry confirmed KIF14 and E2F3 protein overexpression in tumor cells. KIF14 had significant mRNA overexpression with older age (p=0.01) in presenting patients and in unilateral RB patients (p=0.04). Chemotherapy-treated tumors showed a significant decrease in KIF14 and E2F3 expression compared to untreated tumors (p<0.01 and 0.001, respectively). CONCLUSIONS: This report confirms significant mRNA overexpression of KIF14 and E2F3 together in a large cohort of RB tumors. The decreased expression in chemotherapy treated cases needs further validation in a large chemotherapy treated cohort. PMID- 19190781 TI - Reduction and return of infectious trachoma in severely affected communities in Ethiopia. AB - BACKGROUND: Antibiotics are a major tool in the WHO's trachoma control program. Even a single mass distribution reduces the prevalence of the ocular chlamydia that causes trachoma. Unfortunately, infection returns after a single treatment, at least in severely affected areas. Here, we test whether additional scheduled treatments further reduce infection, and whether infection returns after distributions are discontinued. METHODS: Sixteen communities in Ethiopia were randomly selected. Ocular chlamydial infection in 1- to 5-year-old children was monitored over four biannual azithromycin distributions and for 24 months after the last treatment. FINDINGS: The average prevalence of infection in 1- to 5-year old children was reduced from 63.5% pre-treatment to 11.5% six months after the first distribution (P<0.0001). It further decreased to 2.6% six months after the fourth and final treatment (P = 0.0004). In the next 18 months, infection returned to 25.2%, a significant increase from six months after the last treatment (P = 0.008), but still far lower than baseline (P<0.0001). Although the prevalence of infection in any particular village fluctuated, the mean prevalence of the 16 villages steadily decreased with each treatment and steadily returned after treatments were discontinued. CONCLUSION: In some of the most severely affected communities ever studied, we demonstrate that repeated mass oral azithromycin distributions progressively reduce ocular chlamydial infection in a community, as long as these distributions are given frequently enough and at a high enough coverage. However, infection returns into the communities after the last treatment. Sustainable changes or complete local elimination of infection will be necessary. TRIAL REGISTRATION: ClinicalTrials.gov NCT00221364. PMID- 19190784 TI - Capillary microreactors wall-coated with mesoporous titania thin film catalyst supports. AB - A new method for catalyst deposition on the inner walls of capillary microreactors is proposed which allows exact control of the coating thickness, pore size of the support, metal particle size, and metal loading. The wall-coated microreactors have been tested in a selective hydrogenation reaction. Activity and selectivity reach values close to those obtained with a homogeneous Pd catalyst. The catalyst activity was stable for a period of 1000 h time-on-stream. PMID- 19190785 TI - Isotropically etched radial micropore for cell concentration, immobilization, and picodroplet generation. AB - To enable several on-chip cell handling operations in a fused-silica substrate, small shallow micropores are radially embedded in larger deeper microchannels using an adaptation of single-level isotropic wet etching. By varying the distance between features on the photolithographic mask (mask distance), we can precisely control the overlap between two etch fronts and create a zero-thickness semi-elliptical micropore (e.g. 20 microm wide, 6 microm deep). Geometrical models derived from a hemispherical etch front show that micropore width and depth can be expressed as a function of mask distance and etch depth. These models are experimentally validated at different etch depths (25.03 and 29.78 microm) and for different configurations (point-to-point and point-to-edge). Good reproducibility confirms the validity of this approach to fabricate micropores with a desired size. To illustrate the wide range of cell handling operations enabled by micropores, we present three on-chip functionalities: continuous-flow particle concentration, immobilization of single cells, and picoliter droplet generation. (1) Using pressure differentials, particles are concentrated by removing the carrier fluid successively through a series of 44 shunts terminated by 31 microm wide, 5 microm deep micropores. Theoretical values for the concentration factor determined by a flow circuit model in conjunction with finite volume modeling are experimentally validated. (2) Flowing macrophages are individually trapped in 20 microm wide, 6 microm deep micropores by hydrodynamic confinement. The translocation of transcription factor NF-kappaB into the nucleus upon lipopolysaccharide stimulation is imaged by fluorescence microscopy. (3) Picoliter-sized droplets are generated at a 20 microm wide, 7 microm deep micropore T-junction in an oil stream for the encapsulation of individual E. coli bacteria cells. PMID- 19190786 TI - A microfluidic droplet generator based on a piezoelectric actuator. AB - Droplet based microfluidic systems have been shown to be most valuable in biology and chemistry research. However droplet modulation and manipulation requires still further improvement in order to make this technology feasible particularly for biological applications. On demand generation of droplets and droplet synchronization, which is crucial for coalescence, remain largely unanswered. The present study describes a simple and robust droplet generator based on a piezoelectric actuator which is integrated into a microfluidic device. The droplet generator is able to independently control the droplet size, rate of formation and distance between droplets. Moreover, the droplet uniformity is especially high, deviating from the mean value by less than 0.3%. The cross flow and T-junction configurations are tested and show no significant differences, yet the inlet to main channel ratio is found to be important. As this ratio increases, droplets tend to be generated in bursts instead of individually. The physical mechanisms involved are discussed, providing insight into optimized design of such systems. PMID- 19190787 TI - Spectroscopic characterisation and manipulation of arrays of sub-picolitre aerosol droplets. AB - Arrays of optically tweezed aerosol droplets, each of sub-picolitre volume, are manipulated by holographic optical tweezers and characterised by cavity enhanced Raman spectroscopy. A spatial light modulator is employed to generate arrays of optical traps from a single laser beam and to control the array dimensions and relative trap positions. Comparative hygroscopicity measurements are performed concurrently on five trapped droplets by monitoring the evolving size of each droplet. This is extended to the controlled coalescence of an array of droplets accompanied by spectroscopic measurements. These data represent the first ever simultaneous measurements of the evolving composition and size of an array of aerosol droplets. We consider the possibility of using aerosol arrays as a platform for studying chemical reactions in sub-picolitre volumes, exploiting the versatility of aerosol arrays for performing optical digital microfluidic operations accompanied by micro-total analysis. PMID- 19190788 TI - A fluidic device to study directional angiogenesis in complex tissue and organ culture models. AB - Many signals that induce angiogenesis have been identified; however, it is still not clear how these signals interact to shape the vascular system. We have developed a fluidic device for generation of molecular gradients in 3-dimensional cultures of complex tissues and organs in order to create an assay for precise induction and guidance of growing blood vessels. The device features a centrally placed culture chamber, flanked by channels attached to a perfusion system used to generate gradients. A separate network of vacuum channels permits reversible attachment of the device to a flat surface. We show that the fluidic device can be used to create growth factor gradients that induce directional angiogenesis in embryonic mouse kidneys and in clusters of differentiating stem cells. These results demonstrate that the device can be used to accurately manipulate complex morphogenetic processes with a high degree of experimental control. PMID- 19190789 TI - Parallel multi-time point cell stimulation and lysis on-chip for studying early signaling events in T cell activation. AB - Dynamics of complex signaling networks are important to many biological problems. Quantitative data at early time points after cellular stimulation are necessary for accurate model generation. However, the large amount of data needed is often extremely time-consuming and expensive to acquire with conventional methods. We present a two-module microfluidic platform for simultaneous multi-time point stimulation and lysis of T cells for early time point signaling activation with a resolution down to 20 s using only small amounts of cells and reagents. The key design features are rapid mixing of reagents and uniform splitting into eight channels for simultaneous collection of multi-time point data. Chaotic mixing was investigated via computational fluid dynamic modeling, and was used to achieve rapid and complete mixing. This modular device is flexible-with easy adjustment of the setup, a wide range of time points can be achieved. We show that treatment in the device does not elicit adverse cellular stress in Jurkat cells. The activation of six important proteins in the signaling cascade was quantified upon stimulation with a soluble form of alpha-CD3. The dynamics from device and conventional methods are similar, but the microdevice exhibits significantly less error between experiments. We envision this high-throughput format to enable simple and fast generation of large sets of quantitative data, with consistent sample handling, for many complex biological systems. PMID- 19190790 TI - A multipurpose microfluidic device designed to mimic microenvironment gradients and develop targeted cancer therapeutics. AB - The heterogeneity of cellular microenvironments in tumors severely limits the efficacy of most cancer therapies. We have designed a microfluidic device that mimics the microenvironment gradients present in tumors that will enable the development of more effective cancer therapies. Tumor cell masses were formed within micron-scale chambers exposed to medium perfusion on one side to create linear nutrient gradients. The optical accessibility of the PDMS and glass device enables quantitative transmitted and fluorescence microscopy of all regions of the cell masses. Time-lapse microscopy was used to measure the growth rate and show that the device can be used for long-term efficacy studies. Fluorescence microscopy was used to demonstrate that the cell mass contained viable, apoptotic, and acidic regions similar to in vivo tumors. The diffusion coefficient of doxorubicin was accurately measured, and the accumulation of therapeutic bacteria was quantified. The device is simple to construct, and it can easily be reproduced to create an array of in vitro tumors. Because microenvironment gradients and penetration play critical roles controlling drug efficacy, we believe that this microfluidic device will be vital for understanding the behavior of common cancer drugs in solid tumors and designing novel intratumorally targeted therapeutics. PMID- 19190791 TI - An integrated microfluidic culture device for quantitative analysis of human embryonic stem cells. AB - We have successfully designed and fabricated an integrated microfluidic platform, the hESC-microChip, which is capable of reproducible and quantitative culture and analysis of individual hESC colonies in a semi-automated fashion. In this device, a serpentine microchannel allows pre-screening of dissociated hESC clusters, and six individually addressable cell culture chambers enable parallel hESC culture, as well as multiparameter analyses in sequence. In order to quantitatively monitor hESC proliferation and pluripotency status in real time, knock-in hESC lines with EGFP driven by the endogenous OCT4 promoter were constructed. On-chip immunoassays of several pluripotency markers were carried out to confirm that the hESC colonies maintained their pluripotency. For the first time, our studies demonstrated well characterized hESC culture and analysis in a microfluidic setting, as well as a proof-of-concept demonstration of parallel/multiparameter/real-time/automated examination of self-renewal and differentiation in the same device. PMID- 19190792 TI - Cell culture chips for simultaneous application of topographical and electrical cues enhance phenotype of cardiomyocytes. AB - In vivo, cardiomyocytes are exposed to multiple biochemical and physical cues including topographical and electrical cues. During prolonged in vitro cultivation in standard tissue culture set-ups, cardiomyocytes are known to de differentiate due to the lack of appropriate micro-environmental cues. Most currently available cell culture systems provide only a single biophysical cue, thus development of advanced cell cultivation systems incorporating multiple cues is urgently needed. We report here the development of a microfabricated system, incorporating topographical and electrical cues on a single chip, which enables cultivation of differentiated cardiomyocytes. The cell culture chips were created by hot embossing of polystyrene, to create microgrooves and microridges of precisely defined depth, width and periodicity. Substrates consisting of 0.5 microm-wide grooves and 0.5 microm-wide ridges (1 microm period) and those consisting of 3 microm-wide grooves and 1 microm-wide ridges (4 microm period) were investigated, with smooth surfaces used as controls. The depth of the microgrooves was 400 nm. The two gold electrodes were electrodeposited 1 cm apart such that the microgrooves in-between were oriented either parallel or perpendicular to the electrodes, enabling studies of interaction between topographical and electrical cues. Neonatal rat cardiomyocytes cultivated on microgrooved substrates for 7 days were elongated and aligned along the microgrooves forming a well developed contractile apparatus, as evidenced by sarcomeric alpha-actinin staining, with a more pronounced effect on substrates with 1 microm compared to 4 microm periodicity. Importantly, simultaneous application of biphasic electrical pulses and topographical cues resulted in gap junctions confined to the cell-cell end junctions rather than the punctate distribution found in neonatal cells. Electrical field stimulation further enhanced cardiomyocyte elongation when microgrooves were oriented parallel to the electric field. Due to the compatibility of the described cell culture chips with fluorescence and optical microscopy as well as the ability to independently control field stimulation parameters, biochemical and topographical cues on each chip, this system may in the future become a useful tool in drug development and maturation of cardiomyocytes derived from stem cells. PMID- 19190793 TI - The Envirostat - a new bioreactor concept. AB - One major goal of biology is to provide a quantitative description of cellular physiology. This task is complicated by population effects, which perturb culture conditions and mask the behavior of the individual cell. To overcome these limitations, the construction and operation of a microfluidic bioreactor is presented. The new reactor concept guarantees constant environmental conditions and single cell resolution, thus it was named Envirostat (environment, constant). In the Envirostat, cells are contactless trapped by negative dielectrophoresis (nDEP) and cultivated in a constant medium flow. To control chip temperature, a Peltier device was constructed. Joule heating by nDEP was quantified with Rhodamine B in dependence of applied voltage, field mode, medium conductivity, and flow velocity. The integration of the Joule heating effect in the temperature control allowed setting and maintaining the cultivation temperature. For single cell cultivation of Saccharomyces cerevisiae, medium composition changes below 0.001% were estimated by computational fluid dynamic simulation. These changes were considered not to influence cell physiology. Finally, single S. cerevisiae cells were cultivated for more than four generations in the Envirostat, thus showing the applicability of the new reactor concept. The Envirostat facilitates single cell research and might simplify the investigation of hitherto difficult to access biological phenomena such as the true regulatory and physiological response to genetic and environmental perturbations. PMID- 19190794 TI - Massively-parallel ultra-high-aspect-ratio nanochannels as mesoporous membranes. AB - Many fabrication technologies have been used to build nano/mesoporous materials/filters with a good size control, but the integration of these systems into a microsystem format has been a challenge. Microfabricated nanofilters suffer from small open volume and low-throughput. In this paper, we developed a novel fabrication strategy for generating massively-parallel, regular vertical nanochannel membranes with a uniform, well-controlled gap size of approximately 50 nm and a depth up to approximately 40 microm, by using only standard semiconductor fabrication techniques. The vertical nanofilter membranes were fabricated into an anisotropic nanofilter array, which demonstrates the ability to integrate nanofilters and micron-sized channels/pores seamlessly. We demonstrated efficient continuous-flow separation of large DNAs and small molecules in a two-dimensional vertical nanochannel array device. These ultra high-aspect-ratio nanochannels have the advantage of large open volume, enabling high-throughput applications. PMID- 19190795 TI - Microfluidic 2-D PAGE using multifunctional in situ polyacrylamide gels and discontinuous buffers. AB - A two-dimensional microfluidic system is presented for intact protein separations combining isoelectric focusing (IEF) and sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) employing in situ photopolymerized polyacrylamide (PAAm) gels. The PAAm gels are used for multiple functions. In addition to serving as a highly-resolving separation medium for gel electrophoresis, discrete polyacrylamide gel plugs are used to enable the efficient isolation of different on-chip media including anolyte, catholyte, and sample/ampholyte solutions for IEF. The gel plugs are demonstrated as on-chip reagent containers, holding defined quantities of SDS for on-chip SDS-protein complexation, and enabling the use of a discontinuous buffer system for sample band sharpening during SDS-PAGE. The 2-D chip also employs several unique design features including an angled isoelectric focusing channel to minimize sample tailing, and backbiasing channels designed to achieve uniform interdimensional sample transfer. Separation results using E. coli cell lysate are presented using a 10-channel chip with and without the discontinuous buffer system, with resolving power more than doubled in the former case. Further improvements in separation resolution are demonstrated using a 20-channel chip design. PMID- 19190796 TI - Protein self-interaction chromatography on a microchip. AB - This paper presents the development of a novel miniaturized experimental procedure for the measurement of protein-protein interactions through Self Interaction Chromatography (SIC) on a microchip, without the use of chromatographic resins. SIC was recently demonstrated to be a relatively easy method to obtain quantitative thermodynamic information about protein-protein interactions, like the osmotic second virial coefficient B(22), which relates to protein phase behavior including protein crystallization. This successful miniaturization to microchip level of a measurement device for protein self interaction data is a first key step to a complete microfluidic screening platform for the rational design of protein crystallizations, using substantially less expensive protein and experimentation time. PMID- 19190797 TI - A disposable, self-contained PCR chip. AB - A disposable, self-contained polymerase chain reaction (PCR) chip with on-board stored, just-on-time releasable, paraffin-passivated, dry reagents is described. During both storage and sample preparation, the paraffin immobilizes and protects the stored reagents. Fluid flow through the reactor leaves the reagents undisturbed. Prior to the amplification step, the chamber is filled with target analyte suspended in water. Upon heating the PCR chamber to the DNA's denaturation temperature, the paraffin melts and moves out of the way, and the reagents are released and hydrated. To better understand the reagent release process, a scaled up model of the reactor was constructed and the paraffin migration was visualized. Experiments were carried out with a 30 microl reactor demonstrating detectable amplification (with agarose gel electrophoresis) of 10 fg ( approximately 200 copies) of lambda DNA template. The in-reactor storage and on-time release of the PCR reagents reduce the number of needed operations and significantly simplifies the flow control that would, otherwise, be needed in lab on-chip devices. PMID- 19190798 TI - Micropumps operated by swelling and shrinking of temperature-sensitive hydrogels. AB - This paper describes two types of polymeric micropumps based on the temperature sensitive hydrogel poly(N-isopropylacrylamide). The gel actuators are realised as photopolymerised patterns and microgels. They are electrothermically controlled by resistive heating elements. The diffusion-based micropump contains a photopatterned monolithic actuator, which is placed within the pump chamber, and provides a valveless single layer set-up. The diffusion micropump is intended for low performance applications and can operate in two modes: peristaltic or pulsatile. The maximum operating parameters are a flow rate of 2.8 +/- 0.35 microl min(-1) and a back pressure of 1.28 kPa. The second type, a displacement pump, provides a higher performance (maximal 4.5 microl min(-1) and 15 kPa). The pump comprises a microgel-based actuator, which is placed within a separate actuator layer, and active microvalves. The specific features of the design and performance of the pumps are discussed. PMID- 19190799 TI - Platelet function analysis: at the edge of meaning. PMID- 19190800 TI - Thrombin as a common downstream target blocking both platelet and monocyte activation. PMID- 19190801 TI - Using kallikrein-related peptidases (KLK) as novel cancer biomarkers. PMID- 19190802 TI - Thrombotic microangiopathies as a prime example for translational medicine. PMID- 19190803 TI - Autoantibodies in haemolytic uraemic syndrome (HUS). AB - Haemolytic uraemic syndrome (HUS) is a severe disease with renal failure, microangiopathic anemia and thrombocytopenia. Several mechanisms leading to HUS have been identified, like infections with enterohaemorrhagic Escherichia coli, as well as genetic mutations of complement genes, which result in defective complement control on the surface of host cells. The complement system forms the first defense line of innate immunity and mediates the attack against foreign microorganisms. Defective regulation of this cascade results in attack of self cells and in autoimmune disease. Apparently, the alternative pathway convertase C3bBb is central for the pathophysiology of HUS as gene mutations of the components (C3 and Factor B) or of regulators (Factor H, Factor I and MCP/CD46) are observed in the genetic form of HUS. Recently, a novel mechanism leading to atypical HUS (aHUS) was identified, in form of autoantibodies that bind the complement inhibitor Factor H. Here we summarize the current concept of HUS and focus in particular on the novel subgroup of aHUS patients with IgG autoantibodies to Factor H which develop on the genetic background of CFHR1/CFHR3 deficiency, and which define a new subform termed DEAP-HUS (deficient for CFHR proteins and Factor H autoantibody positive). PMID- 19190804 TI - Rituximab as pre-emptive treatment in patients with thrombotic thrombocytopenic purpura and evidence of anti-ADAMTS13 autoantibodies. AB - Thrombotic thrombocytopenic purpura (TTP) is a rare and severe disease characterized by thrombocytopenia, microangiopathic haemolytic anemia, neurological and renal involvement associated with deficiency of the von Willebrand factor-cleaving protease, ADAMTS13. Persistence of high titers of anti ADAMTS13 autoantibodies predisposes to relapsing TTP. Since relapses are associated with high morbidity and mortality rates, the optimal therapeutic option should be a pre-emptive treatment able to deplete anti-ADAMTS13 autoantibodies and avoid relapses. Five patients who presented with persistence of undetectable ADAMTS13 activity and high titers of autoantibodies, were treated with rituximab as pre-emptive therapy during remission. Four of them were affected by relapsing TTP and one was treated after the first episode. ADAMTS13 activity ranging from 15% to 75% with disappearance of inhibitors was achieved after three months in all patients, and persisted >20% without inhibitors at six months. In three patients disease-free status is still ongoing after 29, 24 and six months, respectively. Relapses were documented in two patients during follow up: in one patient remission lasted 51 months; while in the other patient relapse occurred after 13 months. Results demonstrated that rituximab used as pre-emptive treatment may be effective in maintaining a sustained remission in patients with anti-ADAMTS13 antibodies in whom other treatments failed to limit the production of inhibitors, and suggests that re-treatment with rituximab should be considered when ADAMTS13 activity decreases and inhibitors reappear into the circulation, to avoid a new relapse. PMID- 19190805 TI - Variations in the ratio between von Willebrand factor and its cleaving protease during systemic inflammation and association with severity and prognosis of organ failure. AB - Von Willebrand factor (VWF) and related parameters as well as the protease activity regulating its biological activity were measured in plasma of healthy controls and patients with different cause and severity of systemic inflammation to examine the efficacy of the measures to detect highly prothrombotic states including thrombotic microangiopathy (TMA), one of the sequelae of sepsis. Plasma levels of VWF increased with increasing severity of systemic inflammation, probably due to activation of the endothelium. In parallel, the proteolytic activity of VWF inactivating protease, ADAMTS13, stepwise declined with the severity of inflammation, emphasizing the role of VWF-triggered platelet aggregation on the endothelium subsequently followed by development of TMA. As a consequence, the ratio of VWF antigen level and ADAMTS13 activity was significantly higher in patients with inflammation and sepsis, suggesting that this ratio might be more useful for the diagnosis of highly prothrombotic states including TMA than VWF multimer analysis alone. These findings suggest that ADAMTS13, VWF and related parameters, even in a combined approach, might be useful for the diagnosis and the therapeutic monitoring of patients with sepsis associated thrombotic microangiopathy. PMID- 19190806 TI - Pregnancy-associated thrombotic thrombocytopenic purpura. AB - Thrombocytopenia during pregnancy is a common diagnostic and management problem. Several differential diagnosis must be considered including manifestations of thrombotic thrombocytopenic purpura (TTP). We report here on a case of a 21-year old pregnant woman who presented initially severe thrombocytopenia (8 Gpt/l) in the 20(th)+1 week of gestation. The patient had an antibody against ADAMTS13, and enzyme activity was <5%. Immediate plasmapheresis treatment was initiated, followed by plasma infusions, and again plasmapheresis. A male neonate was delivered by caesarean section in the 32(nd )week of gestation. The child had an uncomplicated postnatal development. After delivery, the mother's platelet count and ADAMTS13 activity increased to normal values. This case shows interesting aspects of TTP in pregnancy and a close cooperation between obstetricians, nephrologists and pediatricians is necessary for a successful outcome of the pregnancy. PMID- 19190807 TI - Shiga toxins, glycosphingolipid diversity, and endothelial cell injury. AB - Shiga toxin (Stx)-producing Escherichia coli (STEC) cause an enteric illness that results in a spectrum of outcomes ranging from asymptomatic carriage to uncomplicated diarrhea, bloody diarrhea, and the postdiarrheal haemolytic uremic syndrome (HUS), which leads to renal and other organ microvascular thrombosis. Binding of Stx to the glycosphingolipid (GSL) globotriaosylceramide (Gb3Cer/CD77) on endothelial cells followed by receptor-mediated endocytosis is the linchpin in STEC-mediated disease. Only GSLs that associate strongly with lipid rafts appear to carry Stxs retrogradely from the plasma membrane through the Golgi apparatus to the endoplasmic reticulum where they are translocated to the cytosol and exert their toxic function. Thus, the biophysical features of the lipid moiety of GSL receptors may influence its incorporation into certain membrane domains and thereby affect toxin destination. Consequently, a detailed structural analysis of Stx-binding GSLs is required to illuminate the molecular causes that may underlie the different Stx susceptibilities of endothelial cells derived from various vascular beds. Solid phase overlay binding assays of thin-layer chromatography (TLC)-separated GSL preparations employing specific antibodies and/or Stxs in conjunction with anti-Stx-antibodies are commonly used for the identification of Stx-binding GSLs. Such GSL-profiling combined with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) represents a convenient strategy to structurally characterize Stx-receptors from any biological sources such as primary cells, cell lines, or organs. This approach may be helpful to gain insights into Stx-induced impairment of target cells that is suggested to originate at least partly from the structural heterogeneity of the cellular ligands of Stxs. PMID- 19190808 TI - Pathological aspects of membranoproliferative glomerulonephritis (MPGN) and haemolytic uraemic syndrome (HUS) / thrombocytic thrombopenic purpura (TTP). AB - In this paper, epidemiology, pathogenesis and typical morphological aspects of all three types of membranoproliferative glomerulonephritis (MPGN), of the haemolytic uraemic syndrome (HUS) as well as of thrombotic thrombopenic purpura (TTP) will be reviewed on the light microscopical, immunohistological or immunofluorescence and electron microscopical level. In particular, differences in the pathogenesis of these diseases are discussed. Important recent molecular and genetic insights into the pathogenesis of the three types of MPGN, of typical and atypical HUS and of TTP, i.e. dysregulation of the complement system, distinct molecular defects in C3 and factor H, the major regulatory protein of the alternative pathway of complement activation, and deficiency of a von Willebrand factor (VWF) -cleaving protease, i.e. ADAMTS13, are highlighted. Finally, particular emphasis will be put on differences in glomerular and vascular morphology in the three types of MPGN and in thrombotic microangiopathy (TMA), which is the characteristic morphological alteration of the kidney in HUS and TTP, respectively. PMID- 19190809 TI - Hereditary and acquired complement dysregulation in membranoproliferative glomerulonephritis. AB - Membranoproliferative glomerulonephritis (MPGN) is a chronic progressive renal disease that is diagnosed on the basis of renal histological features. Several MPGN subtypes have been defined by the localization and composition of glomerular deposits (electron dense, Ig and C3). MPGN II or dense deposit disease (DDD) which is defined by the occurrence of electron dense deposits within the lamina densa of the glomerular basement membrane (GBM) is strongly associated with dysregulation of the alternative complement pathway (AP). However, C3 Nephritic Factor (C3NeF), an autoantibody against the alternative C3 convertase C3bBb, and mutations in regulatory proteins of the AP have also been identified in other subtypes of MPGN and even in glomerulonephritis with mesangial C3 deposits. Clinically, MPGN is characterized by proteinuria (up to nephrotic range) and hypertension, frequent progression to end-stage kidney disease and disease recurrence after renal transplantation. The age of onset varies from childhood to adulthood. In the following we will review our current knowledge of pathogenesis of MPGN and will present a novel classification system of the disease based on pathogenesis rather than on morphology. A better understanding of the pathogenesis of MPGN is crucial for the development of novel, specific treatment strategies. PMID- 19190810 TI - Heparin-induced thrombocytopenia: what clinicians need to know. PMID- 19190811 TI - The plasma membrane redox system in human platelet functions and platelet leukocyte interactions. AB - The plasma membrane electron transport is crucial for blood coagulation and thrombosis, since reactive oxygen species and thiol changes, generated by plasma membrane redox reactions, modulate activation of platelets, as well as their interaction with leukocytes. Several antioxidants are linked to this system; thus, platelets are also able to counterbalance radical production and to regulate thrombus growth. Aim of this review is to give an update on the plasma membrane redox system in platelets, as well as on its role in platelet functions and leukocyte-platelet cross-talk. PMID- 19190812 TI - Oral anticoagulant therapy in patients with mechanical heart valve and intracranial haemorrhage. A systematic review. AB - Optimal timing for restarting anticoagulant therapy after intracranial bleeding is a critical issue. The purpose of this systematic review is to summarize published studies on the management of oral anticoagulant therapy after intracranial bleeding secondary to the use of vitamin K antagonists in patients with a mechanical heart valve. A computer-assisted search of the MEDLINE and EMBASE electronic databases till January 2008 was performed. Two investigators independently performed study selection and completed a predefined quality assessment and data extraction form. Main inclusion criterion was the enrolment of patients with a mechanical heart valve and intracranial haemorrhage during oral anticoagulant treatment. Any randomised controlled trial, observational cohort study, case series and reports was included. No randomised controlled trials were identified. Six observational cohort studies were included in the final analysis. All studies were of low quality. A total of 120 patients were enrolled. Anticoagulation was restarted within a broad time range (2 days to 3 months). Four ischaemic strokes and two recurrent cerebral haemorrhages occurred after anticoagulation was restarted after a mean follow-up of 7.9 months. Eighteen patients were described in the selected case reports. Anticoagulant therapy was restarted within four days to eight weeks. Two patients had a recurrent haemorrhagic event, and no ischaemic events were reported. In conclusion, restarting oral anticoagulant therapy few days and, indirectly, stopping anticoagulant therapy for a few days (even for 7-14 days) after the occurrence of cerebral haemorrhage are both safe. However, well-designed studies are strongly recommended to provide better evidence. PMID- 19190813 TI - Microsatellite (GT)(n) repeats and SNPs in the von Willebrand factor gene promoter do not influence circulating von Willebrand factor levels under normal conditions. AB - Von Willebrand factor (VWF) levels vary considerably in normal individuals, influenced by inherited and acquired modulators. ABO blood group is the major inherited determinant of VWF levels, but a role has also been attributed to the VWF gene promoter, haplotype 1 (-3268G/-2709C/-2661A/-2527G) being associated with higher VWF levels than haplotype 2 (-3268C/-2709T/-2661G/-2527A), and the polymorphic locus (GT)(n) modulating the shear stress-induced activation of the VWF promoter. We characterized the (GT)(n) of the VWF promoter in 394 healthy individuals and assessed whether its variable length influenced VWF levels in normal conditions. (GT)(n) proved highly polymorphic, with alleles from 15 to 24 repeats long. (GT)(21) and (GT)(19) were the most common variants (37.4% and 34.4%, respectively). Short GT repeats (15-19) segregated mainly with haplotype 1, long GT repeats (20-24) with haplotype 2 (p < 0.0001). The number of GT repeats did not correlate with VWF levels, nor did such levels correlate with haplotypes 1 and 2, considered alone or in association with the (GT)(n) locus. We conclude that (GT)(n) and -3268/-2709/-2661/-2527 loci are in strong linkage disequilibrium. This polymorphic region of the VWF promoter does not affect VWF levels under normal conditions, though it might represent an environmentally activable VWF regulation site. PMID- 19190814 TI - Increased plasma von Willebrand factor antigen levels but normal von Willebrand factor cleaving protease (ADAMTS13) activity in preeclampsia. AB - The activity of ADAMTS13, the von Willebrand factor (VWF) cleaving protease is low in several conditions, including HELLP (haemolysis, elevated liver enzymes, and low platelet count) syndrome. As HELLP syndrome develops in most cases on the basis of preeclampsia, our aim was to determine whether plasma ADAMTS13 activity is decreased in preeclampsia. Sixty-seven preeclamptic patients, 70 healthy pregnant women and 59 healthy non-pregnant women were involved in this case control study. Plasma ADAMTS13 activity was determined with the FRETS-VWF73 assay, while VWF antigen (VWF:Ag) levels with an enzyme-linked immunosorbent assay. The multimeric pattern of VWF was analyzed by SDS-agarose gel electrophoresis. There was no significant difference in plasma ADAMTS13 activity between the preeclamptic and the healthy pregnant and non-pregnant groups (median [25-75 percentile]: 98.8 [76.5-112.8] %, 96.3 [85.6-116.2] % and 91.6 [78.5 104.4] %, respectively; p > 0.05). However, plasma VWF:Ag levels were significantly higher in preeclamptic patients than in healthy pregnant and non pregnant women (187.1 [145.6-243.1] % versus 129.3 [105.1-182.8] % and 70.0 [60.2 87.3] %, respectively; p < 0.001). The multimeric pattern of VWF was normal in each group. Primiparas had lower plasma ADAMTS13 activity than multi-paras (92.6 [75.8-110.6] % versus 104.2 [92.1-120.8] %; p = 0.011). No other relationship was found between clinical characteristics, laboratory parameters and plasma ADAMTS13 activity in either study group. In conclusion, plasma ADAMTS13 activity is normal in preeclampsia despite the increased VWF:Ag levels. However, further studies are needed to determine whether a decrease in plasma ADAMTS13 activity could predispose preeclamptic patients to develop HELLP syndrome. PMID- 19190815 TI - Factor XIII-A subunit Val34Leu polymorphism is associated with the risk of thrombosis in patients with antiphospholipid antibodies and high fibrinogen levels. AB - Recent reports have described the factor XIII A subunit (FXIII-A) Val34Leu polymorphism as a protective factor against venous and arterial thrombosis. The aim of this study was to investigate the association between the FXIII-A Val34Leu polymorphism, its interaction with fibrinogen concentration, and thrombosis in patients with antiphospholipid antibodies (aPL). We included 172 consecutive patients with aPL: 88 with primary antiphospholipid syndrome (APS), 38 with APS associated with systemic lupus erythematosus (APS-SLE), 32 with SLE and aPL but without APS (SLE-aPL), and 14 asymptomatic individuals with aPL (A-aPL). The FXIII-A Val34Leu polymorphism was assessed by polymerase chain reaction techniques. We found no significant differences in FXIII-A Leu34 allele frequencies between primary APS (allele frequency 0.22), APS-SLE (0.23), SLE-aPL (0.22) and A-aPL (0.32) patients, or between patients with (0.21) and without thrombosis (0.26). FXIII-A Leu34 allele frequencies were significantly lower in patients with thrombosis and those in the upper fibrinogen tertile (>3.40 g/l) (allele frequency 0.07) compared with patients without thrombosis in the upper fibrinogen tertile (0.29) and patients with (0.29) and without (0.25) thrombosis in the mid- and lower fibrinogen tertiles. The FXIII-A Leu34 allele had a protective effect against thrombosis in patients in the upper fibrinogen tertile (odds ratio [OR] = 0.20, 95% confidence interval [CI] 0.07-0.60) but not in those in the other tertiles (OR = 1.20, 95% CI 0.67-2.16). The FXIII-A Leu34 allele seems to have a protective effect on the development of thrombosis in patients with aPL, but only in those with high plasma fibrinogen values. PMID- 19190816 TI - Association study between variants in the fibrinogen gene cluster, fibrinogen levels and hypertension: results from the MONICA/KORA study. AB - Previous studies reported a gender-specific association between plasma fibrinogen concentrations and incident hypertension. We systematically analysed polymorphisms and haplotypes across the fibrinogen gene cluster with fibrinogen levels and assessed their contribution to prevalent hypertension in 2,200 men and 2,159 women from the population-based MONICA/KORA Augsburg study. Eleven tagging single nucleotide polymorphisms (SNPs) were systematically selected in the three fibrinogen genes and haplotypes were reconstructed. The minor alleles of two SNPs, rs2227401 (FGB) and rs2070016 (FGA) and the haplotypes tagged by those variants, were significantly associated with higher fibrinogen concentrations in both, men and women, explaining 1% of the total variance of fibrinogen concentrations. In addition, a FGG haplotype, tagged by rs1049636, was associated with lower concentrations of fibrinogen in women, but not in men. Regarding hypertension, we detected a significant association with a FGA promoter variant (rs2070008) in women only, whereas fibrinogen haplotypes were not associated with hypertension after correction for multiple comparisons in either men or women. In conclusion, our results suggest that variants in all three fibrinogen genes are significantly associated with differences in fibrinogen concentrations with modest contribution to phenotypic variance. It is likely that other genetic variants outside the fibrinogen gene loci are involved in the regulation of fibrinogen concentrations. In addition, one FGA promoter variant was significantly associated with hypertension in women. Confirmation of these findings by future studies is warranted. PMID- 19190817 TI - How useful is determination of anti-factor Xa activity to guide bridging therapy with enoxaparin? A pilot study. AB - Low-molecular-weight heparins (LMWH) are commonly used as peri-procedural bridging anticoagulants. The usefulness of measurement of anti-factor Xa activity (anti-Xa) to guide bridging therapy with LMWH is unknown. It was the objective of this study to determine levels of anti-Xa during standard bridging therapy with enoxaparin, and to examine predictors for residual anti-Xa. Consecutive patients receiving enoxaparin at a dosage of 1 mg/kg body weight/12 hours for temporary interruption of phenprocoumon were prospectively enrolled to the study. Blood samples were obtained 14 hours after LMWH-application immediately pre- procedurally. Procedural details, clinical and demographic data were collected and subsequently analyzed. Seventy patients were included (age 75.2 +/- 10.8 years, Cr Cl 55.7 +/- 21.7ml/min, body mass index [BMI] 27.1 +/- 4.9). LMWH- therapy was for a mean of 4.2 +/- 1.6 days; overall anti-Xa was 0.58 +/- 0.32 U/ml. In 37 (52.8%) of patients anti-Xa was > or U/ml, including 10 (14.3%) patients with anti-Xa > 1U/ml. Linear regression analysis of single variables and logistic multivariable regression analysis failed to prove a correlation between anti-Xa and single or combined factors. No major bleeding, no thromboembolism and four (5.7%) minor haemorrhages were observed. When bridging OAC with therapeutic doses of enoxaparin a high percentage of patients undergo interventions with high residual anti-Xa. The levels of anti-Xa vary largely and are independent of single or combined clinical variables. Since the anti-Xa-related outcome of patients receiving bridging therapy with LMWH is not investigated, no firm recommendation on the usefulness of monitoring of anti-Xa can be given at this stage. PMID- 19190819 TI - Bivalirudin reduces platelet and monocyte activation after elective percutaneous coronary intervention. AB - Concomitant antithrombotic therapy is essential for the prevention of ischaemic events in percutaneous coronary intervention (PCI) and stenting. With new anticoagulant medications being developed and applied in PCI, this raises the question of possible interactions with platelet and leukocyte activation. We therefore sought to investigate the influence of bivalirudin and heparin in platelet and leukocyte activation in patients undergoing elective PCI. Forty-six patients were recruited consecutively in the setting of the Intracoronary Stenting and Antithrombotic Regimen-Rapid Early Action for Coronary Treatment (ISAR-REACT)-3 trial and were randomly assigned to receive either unfactionated heparin or bivalirudin during elective PCI. Surface expression of CD62P (P Selectin), CD42b (GPIbalpha), CD40L, PAC-1 on circulating platelets and CD11b, CD14 and CD15 on circulating leukocytes were evaluated by flow cytometry. Cytokine levels of IL-12p70, tumour necrosis factor (TNF), IL-8, IL-6, IL-1beta and IL-10 were determined by cytometric bead array. Platelet surface expression of PAC-1, P-Selectin and GPIbalpha was significantly reduced after PCI in patients receiving bivalirudin as compared to heparin. Similarly, CD11b expression on CD14+ monocytes was diminished after bivalirudin. However, no differences were observed in cytokine levels between the bivalirudin and the heparin group, before or after PCI. In conclusion, our data suggest that bivalirudin may reduce platelet and monocyte activation in patients undergoing elective PCI. Thereby, bivalirudin might reduce periinterventional thrombotic complications. PMID- 19190818 TI - Comparison of methods to evaluate clopidogrel-mediated platelet inhibition after percutaneous intervention with stent implantation. AB - A high on-treatment residual ADP-inducible platelet reactivity in light transmission aggregometry (LTA) has been associated with an increased risk of adverse outcomes after percutaneous coronary intervention (PCI). However, LTA is weakly standardized, and results obtained in one laboratory may not be comparable to those obtained in another one. We therefore sought to determine the test correlating best with LTA to estimate clopidogrel-mediated platelet inhibition in 80 patients on dual antiplatelet therapy after elective percutaneous intervention with stent implantation. We selected the VerifyNow P2Y12 assay, the vasodilator stimulated phosphoprotein phosphorylation assay, multiple electrode platelet aggregometry and the Impact-R for comparisons with LTA. Cut-off values for residual ADP-inducible platelet reactivity were defined according to quartiles of each assay. Sensitivities and specificities of the different platelet function tests were based on the results from LTA. The results from all four assays correlated significantly with those from LTA. The VerifyNow P2Y12 assay revealed the strongest correlation (r = 0.61, p < 0.001). Sensitivities and specificities ranged from 35% to 55%, and from 78.3% to 85%, respectively. In conclusion, although all assays correlated significantly with LTA, they need to be improved to become clinically used diagnostic tests. Further, it may be too early to define the gold standard method for assessing residual ADP-inducible platelet reactivity and generally acceptable cut-off values. PMID- 19190820 TI - Prolonged exposure to high insulin impairs the endothelial PI3-kinase/Akt/nitric oxide signalling. AB - Hyperinsulinemia predicts future cardiovascular events, but may also contribute to atherosclerosis. We therefore studied the consequences of prolonged insulin treatment of human umbilical vein endothelial cells (HUVEC) on the phosphatidylinositol-3'-kinase(PI3K)/Akt/nitric oxide(NO)-dependent insulin signaling, together with the expression of the pro-atherogenic molecule vascular cell adhesion molecule (VCAM)-1. HUVEC were incubated with insulin (10(-11) to 10(-7) M) in short- (30 min) and long-term (24 h to 3 days) incubations. In short term incubations, insulin did not affect constitutive Akt and eNOS at any concentration, but significantly increased their active phosphorylated forms, and NO production. In long-term incubations, however, such insulin effects on the phosphorylated forms, as well as NO production, were attenuated, promoting an effect of insulin also at concentrations otherwise ineffective. Such effects were accompanied by a boosting of insulin effect on VCAM-1 surface expression. In contrast, under similar conditions, insulin did not exert any significant effect on the surface expression of ICAM-1 and E-selectin. Therefore, prolonged exposure of HUVEC to high insulin levels induces a downregulation of the PI3K/Akt/eNOS axis. Such impairment of insulin signalling in states of prolonged hyperinsulinemia pontially contributes to detrimental effects on atherogenesis in insulin resistance states, such as the metabolic syndrome and type 2 diabetes. PMID- 19190821 TI - Association between polymorphisms in the beta2-adrenergic receptor gene with myocardial infarction and ischaemic stroke in women. AB - Results from studies investigating the association between polymorphisms in the beta2-adrenergic receptor gene (ADRB2) and cardiovascular disease (CVD) are controversial. Using haplotype-based analysis, we have previously shown a protective effect of the Gly16-Gln27-Ile164 haplotype on myocardial infarction in men. We sought to replicate these findings in women and further investigated whether the gene variants exert differential effects on myocardial infarction and ischaemic stroke. We performed a prospective study among 25,224 women, participating in the Women's Health Study and free of CVD at study entry. We had information on polymorphisms Gly16Arg, Gln27Glu, and Thr164Ile in the ADRB2. Incident CVD was self-reported and confirmed after medical record review. We used proportional hazards models to investigate the association between genotypes and haplotypes with any myocardial infarction, any ischaemic stroke, and CVD death. During a mean of 11.8 years of follow-up, 274 myocardial infarctions, 299 ischaemic strokes, and 159 CVD deaths occurred. Among the whole cohort genotype- and haplotype-based analyses did not show an association for any of the gene variants with any of the CVD outcomes. When we focused on Caucasian women, the haplotype-based analysis, however, suggested an inverse association of the haplotype Gly16-Gln27-Thr164 with incident myocardial infarction (multivariable adjusted hazard ratio 0.75; 95% confidence interval 0.58-0.97; p = 0.03). We did not find associations in the haplotype-based analyses with incident ischaemic stroke or CVD death. Our results suggest that the haplotype Gly16-Gln27-Thr164 is associated with reduced risk of incident myocardial infarction but not ischaemic stroke in Caucasian women and suggest differential pathophysiologies for myocardial infarction and stroke. PMID- 19190822 TI - Uncarboxylated matrix Gla protein (ucMGP) is associated with coronary artery calcification in haemodialysis patients. AB - Matrix gamma-carboxyglutamate (Gla) protein (MGP) is a potent local inhibitor of cardiovascular calcification and accumulates at areas of calcification in its uncarboxylated form (ucMGP). We previously found significantly lower circulating ucMGP levels in patients with a high vascular calcification burden. Here we report on the potential of circulating ucMGP to serve as a biomarker for vascular calcification in haemodialysis (HD) patients. Circulating ucMGP levels were measured with an ELISA-based assay in 40 HD patients who underwent multi-slice computed tomography (MSCT) scanning to quantify the extent of coronary artery calcification (CAC). The mean ucMGP level in HD patients (193 +/- 65 nM) was significantly lower as compared to apparently healthy subjects of the same age (441 +/- 97 nM; p < 0.001) and patients with rheumatoid arthritis (RA) without CAC (560 +/- 140 nM; p < 0.001). Additionally, ucMGP levels correlated inversely with CAC scores (r = -0.41; p = 0.009), and this correlation persisted after adjustment for age, dialysis vintage and high-sensitivity C-reactive protein (hs CRP). Since circulating ucMGP levels are significantly and inversely correlated with the extent of CAC in HD patients, ucMGP may become a tool for identifying HD patients with a high probability of cardiovascular calcification. PMID- 19190823 TI - Stroke risk in atrial fibrillation patients on warfarin. Predictive ability of risk stratification schemes for primary and secondary prevention. AB - Atrial fibrillation (AF) patients are widely heterogeneous in terms of ischaemic stroke risk, and several risk stratification schemes have been developed. We performed a prospective study on 662 AF patients on long-term oral anticoagulant therapy (OAT), evaluating the agreement among the different schemes and their correlation with adverse events recorded during follow-up. Patients at low risk were similarly distributed among the different models. Instead, patients classed at moderate risk were 49.2% by CHADS(2) score, 27.6% by NICE and 2.3% by ACCP. As a consequence patients classed at high risk were 46.1% by CHADS(2), 69.8% by NICE and 95.3% by ACCP. CHADS(2 )and NICE scores were associated to the best predictive accuracy. A separate analysis was performed for patients on treatment for secondary prevention, and we observed that they were included in high risk groups by all models, except for 14 patients (6.3%) classed at moderate risk by CHADS(2) even though these patients are at very high risk and the use of aspirin could be unsafe for them. During follow-up 32 major bleeding (1.35 per 100 patient/years) and 39 thrombotic events (1.64 per 100 patient/years) were observed. Among patients on OAT for secondary prevention, both bleeding and thrombotic events mostly occurred in high-risk patients. Even if the absolute rate of adverse events is low, this finding seems to confirm the high stroke risk of this group of patients. For patients on secondary prevention there is no need for further stratification and warfarin should be the treatment of choice. PMID- 19190824 TI - Treatment of PC3 prostate cancer cells with mitoxantrone, etoposide, doxorubicin and carboplatin induces distinct alterations in the expression of kallikreins 5 and 11. AB - Several of the novel kallikrein-related peptidases (tissue kallikreins; KLKs) are emerging new serum and/or tissue biomarkers for prostate cancer (CaP) diagnosis, prognosis and monitoring. In the present research approach, our objective was to investigate the possible alterations in the mRNA expression levels of KLK5 and KLK11 genes in prostate cancer cells PC3 as a response to treatment with mitoxantrone, etoposide, doxorubicin and carboplatin. Viability was assessed with the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay after cell treatment with either mitoxantrone (2 microM), etoposide (20 microM), doxorubicin (1 microM), or carboplatin (15 microM), for 24, 48 and 72 hours. Additionally, trypan blue staining revealed that in PC3 cells all drugs displayed almost the same limited necrotic effects which appeared mainly at 72 hours of treatment. PC3 prostate cancer cells showed a concentration- and time-dependent increased cytotoxicity to the drugs under study which was mainly due to reduction of cell proliferation efficiency. Distinct modulations of KLK5 and KLK11 genes, at the mRNA level, were observed, supporting a drug-dependent cell response. Our experimental data demonstrate that the molecular profile mainly of KLK5 gene may serve as a new potential molecular biomarker predicting treatment response in CaP cells. PMID- 19190825 TI - Expression analysis and study of KLK4 in benign and malignant breast tumours. AB - The steroid hormone-regulated gene KLK4 (kallikrein 4) is a new member of the human kallikrein-related peptidase gene family. Up to date, studies report that KLK4 is differentially expressed in many tumours. The purpose of this study was the expression analysis and study of KLK4 in benign and malignant breast tumours. Total RNA was isolated from 16 benign and 45 malignant breast tissue specimens. After testing RNA quality, cDNA was prepared by reverse transcription. Highly sensitive quantitative real-time PCR method for KLK4 mRNA quantification was developed using the SYBR Green chemistry. GAPDH served as a housekeeping gene. Relative quantification analysis was performed using the comparative C(T) method 2(-DeltaDeltaC)(T) KLK4 expression was found to vary in both patients' cohorts; however, a statistically significant elevation of the KLK4 mRNA levels was observed in malignant compared to benign tumour patients. Low KLK4 expression levels were found in well-differentiated tumours (p = 0.011) as well as in stage I (p = 0.024) patients. Moreover, a statistically significant (r(s) = -0.318, p = 0.035) negative correlation between the KLK4 expression and progesterone receptor staining was observed. ROC and logistic regression analysis recommended that KLK4 gene expression may be used as a new potential biomarker in breast cancer. PMID- 19190826 TI - Effect of plasminogen activator inhibitor-1 on adipogenesis in vivo. AB - To study the functional role of plasminogen activator inhibitor-1 (PAI-1) in obesity, the effect of its overexpression on de novo adipogenesis was evaluated in murine models in vivo. Therefore, 3T3-F442A preadipocytes expressing murine PAI-1 (mPAI-1) or control cells were injected in the back of male NUDE mice, which were fed a high-fat diet (HFD) for four weeks. De novo fat pads that formed from the PAI-1 expressing cells were larger (21 +/- 2.4 mg vs. 14 +/- 1.4 mg; p = 0.017) and showed a higher adipocyte density (373 +/- 28 mm(-2) vs. 301 +/- 12 mm(-2); p = 0.03) as compared to those formed from control cells. In a second model, male NUDE mice were injected in the tail vein with an adenoviral construct expressing mPAI-1 or with the empty vector, and three days later with 3T3-F442A cells. After four weeks of HFD, total body weight and de novo fat pad weight were comparable for both groups. Mild adipocyte hypotrophy was observed in the de novo fat pads of the PAI-1 overexpressing mice (1180 +/- 33 microm(2) vs. 1285 +/- 32 microm(2); p = 0.024), whereas the blood vessel size was significantly smaller than in controls (30 +/- 1.8 microm(2) vs. 63 +/- 3.6 microm(2); p < 0.0001). Thus, the effect of local or systemic PAI-1 (over)expression on adipocyte or blood vessel size and density of de novo formed fat pads appears to be different, and concentration-dependent. Whereas local expression resulted in larger fat pads, systemic overexpression had no effect on de novo adipogenesis, although angiogenesis appeared to be impaired. PMID- 19190827 TI - Point-of-care versus central laboratory coagulation testing during haemorrhagic surgery. A multicenter study. AB - Delay in collecting coagulation test results from a central laboratory is one of the critical issues to efficiently control haemostasis during surgery. The aim of this multicenter study was to compare the performance of a point-of-care (POC) device (CoaguChek Pro DM) with the central laboratory-based coagulation testing during haemorrhagic surgery. For this purpose, 93 patients undergoing major surgical procedure were prospectively included in three centers. Blood was drawn from all patients before surgical incision and from most patients during surgical procedure after a blood loss of 25% or more was observed. When expressed in activity percentage, POC-based prothrombin time (PT) was in good agreement with central laboratory test result with coefficient of correlation in the range from 0.711 to 0.960 in the three centers. Comparison was less conclusive when PT was expressed in seconds or as the patient-to-control ratio and for activated partial thromboplastin time, with significantly shorter clotting times and lower ratios obtained on the POC device. On-site PT (in activity percentage) monitoring would have induced no significant change in fresh frozen plasma (FFP) transfusion in patients when compared to central laboratory monitoring. Test results were obtained in less than 5 minutes when performed using the POC device versus a median turnaround time of 88 minutes (range: 29-235 minutes) when blood collection tubes were sent to the central laboratory. These results suggest that, in providing a rapid answer, POC-based monitoring of PT (in percentage) using the CoaguChek device could be validly used in patients undergoing haemorrhagic surgical procedures. PMID- 19190828 TI - Clinical management of thrombosis in inherited factor VII deficiency: a description of two cases. PMID- 19190829 TI - Activated protein C concentrate treatment for skin necrosis under warfarin treatment in severe genetic protein C deficiency combined with prothrombin mutation and factor V Leiden. PMID- 19190830 TI - Elevation of B cell-activating factor belonging to the tumour necrosis factor [corrected] family (BAFF) in haemophilia A patients with inhibitor. PMID- 19190831 TI - An oral vitamin K protocol to reverse over-anticoagulation in patients presenting with an International Normalised Ratio above 10.0. PMID- 19190832 TI - Respiratory symptoms and conditions related to occupational exposures in machine shops. AB - OBJECTIVES: Since there are few data on the effects of metalworking in populations representing a variety of metal companies or on dose-response relationships concerning metalworking, this study investigated the relationship between occupational exposures in machine shops and the occurrence of upper and lower respiratory symptoms, asthma, and chronic bronchitis. METHODS: A cross sectional study of 726 male machine workers and 84 male office workers from 64 companies was conducted in southern Finland. All of the participants filled out a questionnaire, and aerosol measurements were performed in 57 companies. RESULTS: Exposure to metalworking fluids (MWF) showed a greater risk [odds ratio (OR)>or=2) for upper-airway symptoms, cough, breathlessness, and current asthma than exposures in office work did. Exposure to aerosol levels above the median (>or=0.17 mg/m3 in the general workshop air) was related to an increased risk (OR>or=2) of nasal and throat symptoms, cough, wheezing, breathlessness, chronic bronchitis, and current asthma. Machine workers with a job history of >or=15 years experienced increased throat symptoms, cough, and chronic bronchitis. CONCLUSIONS: This large study representing machine shops in southern Finland showed that machine workers experience increased nasal and throat symptoms, cough, wheezing, breathlessness, and asthma even in environments with exposure levels below the current occupational exposure limit for oil mists. The study suggests that improving machine shop environments could benefit the health of this workforce. It also suggests that it is time to consider reducing the current Finnish occupational exposure limit for oil mist or introducing the use of other health-relevant indicators of exposure. PMID- 19190833 TI - Prevalence and correlates of illicit and non-medical use of psychotropic drugs in Japan: findings from the World Mental Health Japan Survey 2002-2004. AB - PURPOSE: To describe the prevalence of illicit and non-medical use of drugs, the age of first use, and their correlates in the general population of Japan, based on data collected between 2002 and 2004 as part of the World Mental Health (WMH) surveys. METHODS: Participants included were a subsample (n = 887) of the total 2,436 Japanese-speaking respondents aged >or=20 years, randomly sampled from residents in seven cities/municipalities in Japan. Face-to-face household surveys were conducted using the Japanese version of the fully structured WHO WMH Survey Initiative version of the Composite International Diagnostic Interview. RESULTS: Lifetime prevalence of marijuana and cocaine use, non-medical use of prescribed drugs such as tranquilizers, stimulants and analgesics, and use of other substances was 1.5, 0.3, 6.4 and 2.4%, respectively. Lifetime use of marijuana was significantly greater among men. Prescription drug abuse/misuse was significantly more common among the middle-aged (35-49 years) group and those who were married/cohabitating. The 12-month prevalence of marijuana and non-medical use was 0.3 and 1.9%, respectively. Age of first use was likely to be early adulthood. Non-medical use was significantly related to mood disorder, anxiety disorder, intermittent explosive disorder and alcohol abuse/dependence. CONCLUSIONS: The present study confirmed lower prevalence of drug use in Japan than in other countries, such as the United States. However, the non-medical use of psychotropic drugs seems more common in Japan. PMID- 19190834 TI - The persistence of maternal distress and symptoms of distress in adult offspring. AB - This study examined the influence of maternal psychological distress symptoms during offspring's preschool, middle childhood, and adolescent years on the distress symptoms of offspring in adulthood. Data were derived from the British Cohort Study, a longitudinal study of children born in a one-week period in 1970. Results indicated that greater symptoms of maternal distress that persisted over multiple stages of offspring development were generally associated with greater symptoms of distress in adult offspring. The effect of greater maternal distress symptoms that persisted across preschool, middle childhood, and adolescence on adult offspring, however, was not significantly different from the long term effects of a single, but early, exposure to maternal distress once offspring's psychosocial adjustment during adolescence was considered. The relationship between mother and offspring distress symptoms did not vary for male and female offspring. The results provide general support for a cumulative effect perspective in that continuous exposure to maternal distress symptoms had negative consequences in adulthood, and that the adult effect of exposure in early or middle childhood was explained by adolescent adjustment. PMID- 19190835 TI - Impact of early childhood adversities on adult psychiatric disorders: a study of international adoptees. AB - BACKGROUND: This study investigated international adoptees who were taken out of their problematic environments as a consequence of their adoption to determine the effects of early adversities on adult psychiatric disorders, and to study whether these effects emerged de novo after childhood. METHODS: A total of 1,364 adoptees (63.5% of the baseline sample) were followed. Parents provided information about early adversities prior to adoption, and mental health problems in childhood and adolescence. In adulthood, adoptees completed a standardized interview, generating DSM-IV diagnoses. RESULTS: Children who experienced multiple adversities had an increased risk of having anxiety disorders (OR = 2.22; 95% CI: 1.11-4.45), mood disorders (OR = 2.20; 95% CI: 1.00-4.86) or substance abuse/dependence (OR = 3.81; 95% CI: 1.62-8.98) in adulthood. Several effects remained significant after correction for mental health problems in childhood and adolescence. CONCLUSIONS: Severe early adversities increase the risk of adult psychopathology, even when children are taken out of their problematic environments. Results suggest that psychiatric disorders may arise de novo after childhood due to early experiences. PMID- 19190836 TI - Emotional over-involvement can be deleterious for caregivers' health: Mexican Americans caring for a relative with schizophrenia. AB - The study of emotional over-involvement (EOI) has focused primarily on its relationship with patients' course of illness. We know little about the predictors and possible consequences of EOI for caregivers. Based on past research, we tested the hypotheses that EOI is associated with worse physical and psychological health among caregivers and examined whether caregiver burden and social support may mediate this relationship. METHOD: In a sample of 37 Mexican American caregivers and their ill relatives recruited from two outpatient clinics, we examined the relationships between EOI, caregiver burden, caregivers' level of social support, and caregivers' health. Additionally, we examined whether caregiver burden and social support may mediate the relationship between EOI and caregivers' health. Cross-sectional analysis indicates that at baseline EOI was not associated with caregiver burden or social support, but was related to worse current health. Longitudinal analysis, however, indicates that EOI at baseline was associated with greater burden, less instrumental support, and worse health among caregivers at follow-up. Moreover, objective burden and instrumental support mediated the relationship between EOI and several health outcomes. Consequently, EOI may be a marker of poor current health status and predicts worse future health among Mexican-American caregiving relatives of individuals with schizophrenia. Moreover, changes in burden and social support associated with EOI appear to mediate the relationship between EOI and several health outcomes among caregivers. These findings suggest that it might be important for family interventions to not only address the functioning of individuals with schizophrenia but also their caregiving relatives. PMID- 19190837 TI - Isolation of arsenite-oxidizing bacteria from arsenic-enriched sediments from Camarones river, Northern Chile. AB - In Northern Chile, high arsenic concentrations are found in natural water, both natural and anthropogenic sources, a significant health risk. Nine bacterial strains were isolated from Camarones river sediments, located in Northern Chile, a river showing arsenic concentrations up to 1,100 microg/L. These strains were identified as Pseudomonas and they can oxidize arsenite (As(III)) to the less mobile arsenate (As(V)). The arsenite oxidase genes were identified in eight out of nine isolates. The arsenite oxidizing ability shown by the nine strains isolated from arsenic enriched sediments open the way to their potential application in biological treatment of effluents contaminated with arsenic. PMID- 19190838 TI - Use of behavioral responses of rainbow trout Oncorhynchus mykiss in identifying sublethal exposure to hexavalent chromium. AB - Laboratory tests were conducted in a flow-through apparatus on 1-year-old rainbow trout Oncorhynchus mykiss to evaluate the sensitivity of a number of their behavioral responses to hexavalent chromium (Cr(6+)). Test fish were exposed to Cr(6+) concentrations corresponding to 0.001-1 parts of the rainbow trout 96-h LC50 (0.029-28.5 mg Cr/L, respectively) in short-term (15 min) tests. Sensitivity parameter responses could be arranged into the following sequence: latent period of detection response = locomotor activity > gill ventilation frequency > coughing rate. All the rainbow trout responses were sensitive behavioral indicators of sublethal exposure. Behavioral responses meet the criteria as rapid tools for bioassay testing and could be easily standardized using Cr(6+) as a reference toxicant. PMID- 19190839 TI - Seasonal dynamics of phytoplankton in relation to key aquatic habitat factors in a polluted urban small water body in Tianjin, China. AB - To evaluate the seasonal dynamics of phytoplankton and its relationships with aquatic habitat factors in polluted urban landscape lakes, annual investigations have been carried out in the West zone of Xinkai Lake. The results showed that the lake belongs to the eutrophic-supertrophic type in terms of key habitat factors measurement. The seasonal succession of phytoplankton community was determined. The dominant cyanobacterial species Oscillatoria tenuis Ag. bloomed in autumn, although the other species with higher degrees of dominance never bloomed in the year investigated. Significant correlations between some habitat factors and phytoplankton biomass were observed. PMID- 19190840 TI - A preliminary investigation into the effects of X-ray radiation on superficial cranial vascularization. AB - Radiation therapy (RT) is an established treatment modality for malignant neoplasms. RT induces tissue damage that may lead to osteoradionecrosis in more severe cases. Suitable animal models to study RT-induced changes in membranous craniofacial bone are currently not available. The aim of this study was therefore to quantify RT-induced changes in cranial microcirculation using a newly developed calvaria chamber model and to relate these changes to RT-induced histological damage. New Zealand white rabbits received a total radiation dose of 18.75 Gy through the calvaria chamber, and the number of vessels, the vessel length density (VLD), and angiogenic sprouting were quantified on a weekly basis during a 12-week period. At the end of 12 weeks, the RT-treated (n = 5) or control (n = 5) calvarias were biopsied for histopathological analysis. RT resulted in a steep reduction in the number of vessels and the VLD during the first 3 weeks, particularly in larger-diameter vessels, followed by a flat stabilization/remodeling phase in the subsequent 9 weeks that never restored to baseline values. Histomorphometric analysis revealed a high degree of osteocytic depletion, prominent hypocellularity in the lacunae and intraosseous vasculature, enlarged and nonconcentric Haversian systems, and a severely disorganized bone matrix in the RT-treated calvarias. Despite the prevalence of some angiogenic potential, the RT-induced effects in the early phase persisted in the intermediate to late phase, which may have contributed to the poor recovery of the RT-treated bone. PMID- 19190843 TI - Personalized models of bones based on radiographic photogrammetry. AB - The radiographic photogrammetry is applied, for locating anatomical landmarks in space, from their two projected images. The goal of this paper is to define a personalized geometric model of bones, based uniquely on photogrammetric reconstructions. The personalized models of bones are obtained from two successive steps: their functional frameworks are first determined experimentally, then, the 3D bone representation results from modeling techniques. Each bone functional framework is issued from direct measurements upon two radiographic images. These images may be obtained using either perpendicular (spine and sacrum) or oblique incidences (pelvis and lower limb). Frameworks link together their functional axes and punctual landmarks. Each global bone volume is decomposed in several elementary components. Each volumic component is represented by simple geometric shapes. Volumic shapes are articulated to the patient's bone structure. The volumic personalization is obtained by best fitting the geometric model projections to their real images, using adjustable articulations. Examples are presented to illustrating the technique of personalization of bone volumes, directly issued from the treatment of only two radiographic images. The chosen techniques for treating data are then discussed. The 3D representation of bones completes, for clinical users, the information brought by radiographic images. PMID- 19190842 TI - Bone turnover markers: understanding their value in clinical trials and clinical practice. AB - While bone mineral density (BMD) by dual-energy X-ray absorptiometry is the primary method of determining fracture risk, assessing bone turnover may add valuable information for the management of patients with low bone mass. Bone turnover markers (BTMs) are used in clinical trials where they can provide essential information on the biological efficacy of osteoporosis treatments. In such population-based studies, BTMs can predict fracture risk independent of BMD. When combined with BMD, they improve the fracture risk estimate above and beyond BMD alone in postmenopausal osteoporotic women. Since changes in bone turnover after the initiation of therapy with bone resorption inhibitors occur much more rapidly than changes in BMD, treatment efficacy could, in theory, be determined within weeks of using BTMs. However, such predictive value is limited by the large biological variability of these biochemical markers, even though newer automated methods have reduced their analytical variability. Consequently, widespread adoption as a means of predicting treatment efficacy in fracture prevention for individual patients cannot yet be recommended. BTMs may be useful for monitoring adherence to antiresorptive therapy and may aid in identifying patients for whom antiresorptive therapy is most appropriate. Thus, although BTMs are currently confined to clinical research applications, further improvement in assay precision may extend their diagnostic value in clinical settings. PMID- 19190844 TI - Spatial relationships between lingual nerve and mandibular ramus: original study method, clinical and educational applications. AB - Lingual nerve damage complicating oral surgery would sometimes require electrographic exploration. Nevertheless, direct recording of conduction in lingual nerve requires its puncture at the foramen ovale. This method is too dangerous to be practiced routinely in these diagnostic indications. The aim of our study was to assess spatial relationships between lingual nerve and mandibular ramus in the infratemporal fossa using an original technique. Therefore, ten lingual nerves were dissected on five fresh cadavers. All the nerves were catheterized with a 3/0 wire. After meticulous repositioning of the nerve and medial pterygoid muscle reinsertion, CT-scan examinations were performed with planar acquisitions and three-dimensional reconstructions. Localization of lingual nerve in the infratemporal fossa was assessed successively at the level of the sigmoid notch of the mandible, lingula and third molar. At the level of the lingula, lingual nerve was far from the maxillary vessels; mean distance between the nerve and the anterior border of the ramus was 19.6 mm. The posteriorly opened angle between the medial side of the ramus and the line joining the lingual nerve and the anterior border of the ramus measured 17 degrees . According to these findings, we suggest that the lingual nerve might be reached through the intra-oral puncture at the intermaxillary commissure; therefore, we modify the inferior alveolar nerve block technique to propose a safe and reproducible protocol likely to be performed routinely as electrographic exploration of the lingual nerve. What is more, this original study protocol provided interesting educational materials and could be developed for the conception of realistic 3D virtual anatomy supports. PMID- 19190845 TI - Sulcal and gyral anatomy of the orbitofrontal cortex in relation to the recurrent artery of Heubner: an anatomical study. AB - The aim of this study is to investigate the sulcal and the gyral anatomy of the orbitofrontal cortex with its arterial supply. Ten gross formaline fixed adult brains (20 hemispheres) were used to show the gyral and sulcal anatomy of the inferior frontal lobe, and its arterial supply. The arteries were investigated with special attention to the relationship between the recurrent artery of Heubner (RAH) and the gyrus rectus (GR). Medial and lateral orbital sulci were connected in 4 right hemispheres, and in 7 of the left. The orbital sulci were connected with olfactory sulcus in one right hemispheres, but not in the left. In the right hemispheres, the RAH traveled across the GR in 7 hemispheres and looped over the posterior aspect of the GR in 3 hemispheres. In the left hemispheres, RAH crossed the GR in 8 and looped over the posterior aspect of the GR before entering the anterior perforating substance in 2 hemispheres. There are considerable variations among the gyri and sulci of the orbitofrontal cortex and it is difficult to describe a precise architectural pattern. The RAH demands special attention during GR resection in aneurysm surgery. PMID- 19190846 TI - Microsurgical anatomy of the foramen of Luschka in the cerebellopontine angle, and its vascular supply. AB - Cerebellopontine angle and vascular supply of adjacent brainstem and cerebellum are susceptible to compression and eventual damage by tumors. Delicate and complicated neurosurgical operations in the cerebellopontine angles of the brainstem, where lateral recesses of fourth ventricle empty, are abundant especially operations in which foramina of Luschka are used as possible access to the floor of the fourth ventricle. So awareness and knowledge of the normal anatomical features of the region is valuable for neurosurgeons. Arteries of 40 human cerebella were injected with colored gelatin to investigate the microsurgical anatomy around the foramen of Luschka in the cerebellopontine angle. Two compartments of the foramen of Luschka were distinguished, choroidal part and the patent part. Seventy-four (92.5%) of foramina were open and only 6 (7.5%) foramina were closed. The mean distance between the foramen of Luschka and the anterior inferior cerebellar artery was 3.90 mm on the left side and 3.89 on the right side. The distance from the posterior inferior cerebellar artery was 7.08 and 5.81 mm to the left and right foramina of Luschka, respectively. In ten cases, tortuous vertebral artery was occupying the left cerebellopontine angle space and the foramen of Luschka. PMID- 19190847 TI - Oblique multiplanar reformation in multislice temporal bone CT. AB - We made an anatomic study using a convenience sample of 20 patients, most of them referred to our institution for depicting internal auditory malformations that justify sensorineural deafness or for surgical planning of cochlear implants. All patients underwent a multislice temporal bone CT and oblique single slice reformation postprocessing in six proposed different planes corresponding to cochlear basal turn, apical basal turn, malleoincudal complex, stapes, and facial channel. Anatomic and pathologic characterization of some middle and inner ear structures, difficult to evaluate in standard axial and coronal planes, can be improved using this technique. PMID- 19190848 TI - Percutaneous cervical disc decompression. AB - BACKGROUND: Cervical disc nucleoplasty is a significant and clinically demonstrated innovation in percutaneous disc decompression in case of non herniated disc protrusions or prolpase. It allows a percutaneous decompression via a 19-gauge needle under utilization of the Coblation technique and under C arm control. Until now the patients suffering of a cervicobrachialgia in cause of a disc prolapse had only the therapeutical solution between conservative treatment and monosegmental spondylodesis or disc prosthesis of the mentioned motion segment. METHODS: We wanted to demonstrate a new and practicable anatomical pathway for reaching the cervical disc prolapse comparable to the technique for discography of the cervical spine. The introducer needle is advanced into the disc under fluoroscopic guidance using a standard anterior lateral approach. The controller delivers radiofrequency energy to quickly ablate tissue at temperatures between 50 degrees and 60 degrees C. The decompression will be done in ablation mode by rotating the device through 180 degrees for 5 s in the posterior, medial and ventral third of the cervical disc. After failed conservative treatment over an average time period of 3 months we treated 26 patients with a contained herniated prolapse or protrusion with radicular arm pain by percutaneous decompression under utilization of the Coblation technique with a controlled energy plasma-mediated field. A randomized control group of 30 patients was treated alone conservatively with medical and physical therapy in the same period. RESULTS: The average preoperative VAS was 8.8. With a follow-up time of 2-years we found an average pain reduction with the visual pain score (VAS) of 2.3 who had a further check-up. The VAS was checked 24 h, 1 week, 3, 6, 12 and 24 months postoperatively. No complications with this method were seen. Comparable to the surgically treated group the conservative patients have had a VAS of 8.4. Under using conservative treatment with physical therapy, physiotherapy, analgetics and perineural injections we have had a diminution of the VAS to 5.1 after 2 years. CONCLUSION: The percutaneous decompression of the cervical disc protrusion with the Perc DC-Spine Wand by using the Coblation mode is a quick and safe procedure. Furthermore, one may state a persistent pain relief in the follow-up time up to 2 years after the percutaneous decompression of the disc. PMID- 19190849 TI - Clinical anatomy study of autonomic nerve with respective to the anterior approach lumbar surgery. AB - INTRODUCTION: Male genital dysfunction was recognized as a complication following anterior approach lumbar surgery. Disruption of efferent sympathetic pathways such as the abdominal aortic plexus (AAP) and superior hypogastric plexus (SHP) which lied pre-abdominal aorta and iliac artery had been thought as the main reason. Though there were some clinical reports of retrograde ejaculation, the applied anatomic study of the autonomic nerve anterior to the lumbar was little. The purpose was to find out a lumbar surgery approach which was ejaculation preservation through the detailed study of the anatomy and histology observation of the autonomic nerve anterior to the lumbar vertebrae. METHODS: The lumbar region of ten male cadavers was dissected and analyzed. We investigated the relationship between the peritoneum and abdominal aorta, iliac artery and sacral promontory fascia, as well as the trend and distribution of the autonomic nerve and SHP anterior to the L5-S1. We also observed the distribution of autonomic nerve at retroperitoneum through hematoxylin and eosin (HE)-stained tissues pre aorta, para-aorta, and pre-vertebrae sacrales. RESULTS: Superior hypogastric plexus, which deviated to left, located in a triangle formed by the common iliac arteries and its bilateral branches, its truck sited anterior to the lumbarsacral space in seven cases (70%), and anterior to sacrum in three cases (30%); at the aortic bifurcation, SHP strided over left iliac artery from left-hand side, then located in front of sacrum in four cases (40%), and sifted to the left at the lumbar sacral promontory in six cases (60%); from both anatomic and histological view, the autonomic nerve plexus lying in an fascia layer of retroperitoneum. CONCLUSION: At the anterior approach lumbar surgery of trans-peritoneum, we should choose the right-hand side incision; the SHP should be pushed aside carefully from right to left along intervertebral disc. The accurate surgical plane was at the deeper layer of autonomical nerve fascia; we also could lift the complete autonomical nerve layer which lies behind the aorta and lumbar sacral promontory, so that the autonomic nerve could be preserved. PMID- 19190850 TI - Connection types between the spinal root of the accessory nerve and the posterior roots of the C2-C6 spinal nerves. AB - PURPOSE: The aim of this study was to demonstrate the connection types and frequency between the accessory nerve and the posterior roots of the C2-C6 cervical nerves. METHODS: The cranial cervical regions of 49 specimens from 27 human cadavers were used for the present study under an operating microscope. RESULTS: Five different connection types between the accessory nerve and the posterior roots of the cervical nerves were recorded and photographed (types A F). One of these types was not described previously in literature (type F). All connections between the posterior roots of the C2-C6 spinal nerves and the accessory nerve were at the level of the C2 segment. Type B was the most frequently seen type in our series. One of the rootlets of the cervical posterior root joined the accessory nerve without a connection to the spinal cord in type B. CONCLUSIONS: The clinical importance of these connections is especially noticed during the radical neck dissection as it may lead to the development of the shoulder-arm syndrome. PMID- 19190851 TI - The course of the inferior gluteal nerve and surgical landmarks for its localization during posterior approaches to hip. AB - The position of the inferior gluteal nerve (IGN) makes it vulnerable to iatrogenic injury during posterior and posterolateral approaches to the hip. Although the posterior approach has been reported to be the most frequently used technique, it is most likely to be associated with damage to the IGN. As there is scant information in the literature regarding the course and the anatomic relationships of the IGN, we aimed to investigate the anatomic course of the IGN and define the anatomical landmarks that can be used by surgeons during posterior approaches to the hip. Thirty-six gluteal regions from adult fixed cadavers were used for this study. A triangular-shaped anatomic area that contains the IGN was defined. This geometric area was formed by connecting the following points: posterior inferior iliac spine (PIIS) (apex), ischial tuberosity (IT) and greater trochanter (GT). This triangle can further be divided into two, the upper triangle being the "danger zone" since it contains the IGN and its branches. The closest mean distance between the point of IGN origin and the PIIS, IT and the GT was 3.2, 4.8 and 5.4 cm, respectively. In all specimens, the nerve entered the deep surface of the gluteus maximus approximately 5.4 cm from the apex of the GT and approached the GT as close as 0.8 cm, on average. Based on our study, dividing the gluteus maximus with standard techniques may damage the IGN. Posterior minimally invasive approaches to the hip should take into account the point of entry of the IGN into the gluteus maximus. Localization of the IGN by using the anatomic triangle defined in this study may decrease surgical morbidity. PMID- 19190853 TI - Dilemmas caused by endogenous pararetroviruses regarding the taxonomy and diagnosis of yam (Dioscorea spp.) badnaviruses: analyses to support safe germplasm movement. AB - The discovery of endogenous pararetroviral sequences (EPRVs) has had a deep impact on the approaches needed for diagnosis, taxonomy, safe movement of germplasm and management of diseases caused by pararetroviruses. In this article, we illustrate this through the example of yam (Dioscorea spp.) badnaviruses. To enable progress, it is first necessary to clarify the taxonomical status of yam badnavirus sequences. Phylogeny and pairwise sequence comparison of 121 yam partial reverse transcriptase sequences provided strong support for the identification of 12 yam badnavirus species, of which ten have not been previously named. Virus prevalence data were obtained, and they support the presence of EPRVs in D. rotundata, but not in D. praehensilis, D. abyssinica, D. alata or D. trifida. Five yam badnavirus species characterised by a wide host range seem to be of African origin. Seven other yam badnavirus species with a limited host range are probably of Asian-Pacific origin. Recombination under natural circumstances appears to be rare. Average values of nucleotide intra species genetic distances are comparable to data obtained for other RNA and DNA virus families. The dispersion scenarios proposed here, combined with the fact that host-switching events appear common for some yam badnaviruses, suggest that the risks linked to introduction via international plant material exchanges are high. PMID- 19190855 TI - Telopeptides of type II collagen upregulate proteinases and damage cartilage but are less effective than highly active fibronectin fragments. AB - OBJECTIVE AND DESIGN: We hypothesize that N-telopeptide (NT) and C-telopeptides (CT) of type II collagen can enhance proteinases and cause cartilage damage and have compared damaging activities to an extensively characterized potent fibronectin fragment (Fn-f). MATERIALS: NT and CT peptides were synthesized. METHODS: Interaction of labeled peptides with chondrocytes was studied by fluorescence microscopy. Effects on the metalloproteinases (MMPs) MMP-3 and MMP 13 and on ADAMTS-5 were analyzed by western blotting. Cartilage damage was assayed by loss of proteoglycan (PG) from cultured explants. RESULTS: NT and CT peptides penetrated cartilage, bound to chondrocytes and enhanced proteinase release and cartilage PG depletion. Peptides had detectable activity at 0.3 microM (1 microg/ml) and were comparable at 30 microM (100 microg/ml) to 1 microM Fn-f (29 microg/ml). However, while the Fn-f enhanced IL-1beta and TNF-alpha, the NT and CT peptides did not. CONCLUSIONS: Collagen peptides containing NT and CT regions were less active on a molar basis than Fn-fs but were still potent damaging agents. Since collagen fragments are found in OA cartilage at microg/ml, they have the potential to play a role in physiologic cartilage damage. PMID- 19190856 TI - CD40/CD154 system and pro-inflammatory cytokines in young healthy male smokers without additional risk factors for atherosclerosis. AB - OBJECTIVE AND DESIGN: Atherosclerosis, as an inflammatory disease, is characterized by pathologically altered levels of cytokines. We investigated whether smoking affects the CD40/CD154 system and pro-inflammatory cytokines in young males without other risk factors for atherosclerosis. SUBJECTS: Young male smokers (n=13) and 14 non-smoking controls were investigated. METHODS: The differences in CD40/CD154 system and serum cytokines between the groups were measured using flow cytometry and ELISA. RESULTS: In smokers, there was a strong trend (P<0.06) for increased CD40 expression on platelets as compared with non smokers. However, there were no significant differences in CD40 expression on monocytes or in CD154 expression on platelets and T-cells between smokers and non smokers. There was a strong trend for increased platelet-monocyte aggregates in smokers (P<0.06). Also, smokers had slightly but not significantly elevated hsCRP and IL-6 levels, and slightly decreased TNF-alpha and MCP-1. Interestingly, IL 18, a cytokine which has the ability to promote both Th1 and Th2 responses, was significantly decreased in smokers group (P=0.03 vs controls). CONCLUSIONS: In young healthy males, smoking is not associated with dramatic changes in CD40/CD154 system. However, cigarette smoke alters the secreted cytokine profile, leading to significant decrease in systemic IL-18 levels. PMID- 19190858 TI - Eye injury (ocular trauma) in southern Turkey: epidemiology, ocular survival, and visual outcome. AB - PURPOSE: To evaluate the epidemiology and outcome of ocular trauma in southern Turkey. METHODS: Patients referred to the University of Cukurova, School of Medicine, Department of Ophthalmology, with ocular trauma during the last two years were reviewed retrospectively. Age and gender of patients, type of injury, cause of injury, place where injury occurred, initial and final visual acuities, and final outcome were analyzed. RESULTS: Medical records of 250 patients (four bilateral injuries), aged between three months and ninety years were reviewed. Ocular trauma mostly occurred between 0 and 15 years and a male preponderance (76.8%) was evident. There were 72% open-globe injuries, corneal and/or scleral penetration being the most common injury type. Metallic objects were the main cause of injury (32.4%). Fifty percent of injuries happened in streets. Closed globe and self-sealing corneal injuries were found to have the best prognosis in terms of final outcome (57.6% and 57.9%, respectively; P < 0.001 for both). Initial visual acuity was found to be a significant predictor of final visual acuity (P < 0.001). Endophthalmitis was significantly associated with metallic objects (P < 0.005). CONCLUSION: Eye injury is still a common and avoidable cause of blindness. Our findings suggest that any person in the population can be subject to ocular trauma at any time. Prognosis might be challenging. Education, attention, and safety precautions would help in reducing ocular morbidity. PMID- 19190857 TI - A clinical approach to the diagnosis of retinal vasculitis. AB - Retinal vasculitis is a sight-threatening inflammatory eye condition that involves the retinal vessels. Detection of retinal vasculitis is made clinically, and is confirmed with the help of fundus fluorescein angiography. Active vascular disease is characterized by exudates around retinal vessels resulting in white sheathing or cuffing of the affected vessels. In this review, a practical approach to the diagnosis of retinal vasculitis is discussed based on ophthalmoscopic and fundus fluorescein angiographic findings. PMID- 19190859 TI - Short and long-term benefits of sirolimus-eluting stent in ST-segment elevation myocardial infarction: a meta-analysis of randomized trials. AB - BACKGROUND: Recent concerns have emerged on the potential higher risk of stent thrombosis after DES implantation, that might be even more pronounced among STEMI patients. The aim of the current study was to perform a meta-analysis to evaluate the benefits and safety of Sirolimus-Eluting Stent (SES) as compared to BMS in patients undergoing primary angioplasty for STEMI. METHODS: The literature was scanned by formal searches of electronic databases (MEDLINE and CENTRAL). We examined all completed randomized trials of DES for STEMI. The following keywords were used for study selection: randomized trial, myocardial infarction, reperfusion, primary angioplasty, stenting, DES, sirolimus-eluting stent (SES), Cypher. Information on study design, type of stent, inclusion and exclusion criteria, primary endpoint, number of patients, angiographic and clinical outcome, were extracted by two investigators. Disagreements were resolved by consensus. RESULTS: A total of 9 trials were included in the meta-analysis, involving 2,769 patients (1389 or 50.2% randomized to DES and 1,380 or 49.8% randomized to BMS). At 12 months follow-up, SES was associated with a significant reduction in TVR (4.9% vs. 13.6%, p < 0.0001), with a trend in benefits in mortality (2.9% vs. 4.2%, p = 0.08) and reinfarction (3.0% vs. 4.3%, p = 0.06), without any significant difference in stent thrombosis (1.9% vs. 2.5%, p = 0.36). Safety and efficacy of DES were confirmed at 2-3 years follow-up (data available from 4 trials including 569 patients). CONCLUSIONS: This meta-analysis shows that among selected STEMI patients undergoing primary angioplasty, SES as compared to BMS is safe and associated with a significant reduction in TVR at 1 and 2-3 years follow-up. PMID- 19190860 TI - Multiple responses optimization and modeling of lipase production by Rhodotorula mucilaginosa MTCC-8737 using response surface methodology. AB - Response surface methodology was employed to optimize culture medium for production of lipase with Rhodotorula sp. MTCC 8737. In the first step, a Plackett-Burman design was used to evaluate the effects of different inducers qualitatively. Of all the seven inducers tested, soybean oil showed significant influence on the lipase production. Further, response surface studies were conducted to quantitatively optimize by considering linear, interactive, and quadratic effects of test variables. A novel approach was proposed to optimize the lipase production system by optimizing the responses in terms of yield kinetics rather than optimizing the direct responses like lipase titer and biomass growth. The coefficient of determination (R(2)) calculated for Y (P/S) (0.769), Y (P/X) (0.799), and Y (X/S) (0.847) indicated that the statistical model could explain 76.9%, 79.99%, and 84.7% of variability in the response. PMID- 19190861 TI - Salicylic-acid-mediated enhanced biological treatment of wastewater. AB - Activated sludge represents a microbial community which is responsible for reduction in pollution load from wastewaters and whose performance depends upon the composition and the expression of degradative capacity. In the present study, the role of salicylic acid (SA) has been evaluated for acclimatization of activated sludge collected from a combined effluent treatment plant followed by analysis of the physiological performance and microbial community of the sludge. The biodegradative capacity of the acclimatized activated sludge was further evaluated for improvement in efficiency of chemical oxygen demand (COD) removal from wastewater samples collected from industries manufacturing bulk drugs and dyes and dye intermediates (wastewater 1) and from dye industry (wastewater 2). An increase in COD removal efficiency from 50% to 58% and from 78% to 82% was observed for wastewater 1 and wastewater 2, respectively. Microbial community analysis data showed selective enrichment and change in composition due to acclimatization by SA, with 50% of the clones showing sequence homology to unidentified and uncultured bacteria. This was demonstrated by analysis of partial 16S rDNA sequence data generated from dominating clones representing the metagenome which also showed the appearance of a unique population of clones after acclimatization, which was distinct from those obtained before acclimatization and clustered away from the dominating population. PMID- 19190862 TI - Evaluation of cashew apple juice for surfactin production by Bacillus subtilis LAMI008. AB - Bacillus subtilis LAMI008 strain isolated from the tank of Chlorination at the Wastewater Treatment Plant on Campus do Pici in Federal University of Ceara, Brazil has been screened for surfactin production in mineral medium containing clarified cashew apple juice (MM-CAJC). Results were compared with the ones obtained using mineral medium with glucose PA as carbon source. The influence on growth and surfactin production of culture medium supplementation with yeast extract was also studied. The substrate concentration analysis indicated that B. subtilis LAMI008 was able to degrade all carbon sources studied and produce biosurfactant. The highest reduction in surface tension was achieved with the fermentation of MM-CAJC, supplemented with yeast extract, which decreased from 58.95 +/- 0.10 to 38.10 +/- 0.81 dyn cm(-1). The biosurfactant produced was capable of emulsifying kerosene, achieving an emulsification index of 65%. Surfactin concentration of 3.5 mg L(-1) was obtained when MM-CAJC, supplemented with yeast extract, was used, thus indicating that it is feasible to produce surfactin from clarified cashew apple juice, a renewable and low-cost carbon source. PMID- 19190863 TI - Purification and characterization of thermostable chitinase from Bacillus licheniformis SK-1. AB - Chitinase was purified from the culture medium of Bacillus licheniformis SK-1 by colloidal chitin affinity adsorption followed by diethylamino ethanol-cellulose column chromatography. The purified enzyme showed a single band on sodium dodecyl sulfate polyacrylamide gel electrophoresis. The molecular size and pI of chitinase 72 (Chi72) were 72 kDa and 4.62 (Chi72) kDa, respectively. The purified chitinase revealed two activity optima at pH 6 and 8 when colloidal chitin was used as substrate. The enzyme exhibited activity in broad temperature range, from 40 to 70 degrees C, with optimum at 55 degrees C. It was stable for 2 h at temperatures below 60 degrees C and stable over a broad pH range of 4.0-9.0 for 24 h. The apparent K (m) and V (max) of Chi72 for colloidal chitin were 0.23 mg ml(-1) and 7.03 U/mg, respectively. The chitinase activity was high on colloidal chitin, regenerated chitin, partially N-acetylated chitin, and chitosan. N bromosuccinamide completely inhibited the enzyme activity. This enzyme should be a good candidate for applications in the recycling of chitin waste. PMID- 19190864 TI - [Delirium in the old age--still inadequately recognised and treated]. PMID- 19190866 TI - [Delirium--management in the hospital: diagnosis and treatment]. AB - All states of confusion with acute onset and alteration of thinking, perception and awareness are defined as "delirium". Delirium is a common problem in older patients admitted to the hospital. It is combined with a negative prognosis and complications (falls, infections, etc.). Diagnosis and management need special efforts. Delirium is mostly associated with an underlying disease. In the elderly patient, nearly every disease can be accompanied by delirium, and the fragile older patient is especially affected.Delirium should be evaluated using a systematic approach according to frequency and impact of possible causes. An accurate diagnosis can be challenging. Symptoms have to be distinguished from other cerebral alterations, such as preexisting dementia or depression. Due to the high prevalence, one should always be aware to the presence of delirium.With the treatment of the identified disease, there is generally a reduction of delirium. Often medical intervention in the acute state is necessary to prevent further alteration of the patient. Beside an adequate medical strategy, it is important to create a safe environment for the affected patient. The article gives suggestions for a systematic diagnostic and therapeutic strategy of delirium. PMID- 19190865 TI - [Delirium in the elderly]. AB - Delirium is a common, serious medical and often life-threatening condition in elderly in-patients. Delirium can develop primarily or secondarily related to other medical conditions and lead to hospital admission. The pathogenesis is still not fully known and is usually addressed as multifactorial. Alterations in neurotransmitters have a key role in this process. The incidence varies by setting up to 90%. Delirium is associated with increased short- and long-term mortality, iatrogenic complications, functional decline, and future development of cognitive impairment or dementia. Delirium is also associated with longer hospital stays, higher hospital and total health system costs, and an increasing rate of nursing home admissions. A structured diagnostic and therapeutic process is recommended. Delirium should become a quality indicator for hospital medicine; however, many research questions still exist. PMID- 19190867 TI - How to prevent perioperative delirium in the elderly? AB - Due to increasing life expectancy, the number of elderly patients needing surgical care is increasing. Improvements in surgical techniques and anesthetic procedures offer the opportunity of surgical intervention even in frail patients. Delirium on admission to the hospital or in the perioperative setting is a common and often serious complication. Cognitive impairment is regarded as the main risk factor for delirium; unfortunately, under routine clinical conditions, these deficits often remain undetected. Postoperative delirium is associated with increased morbidity and mortality, as well as increased length of hospitalization, resulting in increased suffering and costs. The aim of the intervention presented here was to prevent delirium in a general hospital without a geriatric specialty department. Geriatric nurses became part of the team in the perioperative setting, giving psychological support and being a continuous companion to the patient and his/her proxies. Co-operation between all professions involved throughout hospitalization of the patient was emphasized. The low incidence of delirium in elderly surgical patients resulting from our efforts indicate that - for this setting in a general hospital - the strategy was effective for the prevention of delirium. Nowadays, geriatric nurses are an essential part of the perioperative team and delirium has lost much of its imminence. PMID- 19190868 TI - A special care unit for acutely ill patients with dementia and challenging behaviour as a model of geriatric care. AB - This paper describes the development and management of a new model of care for hospitalized patients with challenging behaviour evoked by dementia and/or delirium. To ameliorate care for patients with dementia in a geriatric acute care hospital a segregated Special Care Unit for patients with challenging behaviour was created. Environmental features allow for safe and unrestricted ambulation within the unit and create a home-like atmosphere. Day-time activities structure the day and assure additional professional presence in the unit. An intensive training program for the staff was provided. The SCU has been well accepted by the staff and is considered to be an improvement in care. Psychological burden of the nurses did not increase over a time period of one year in caring for these difficult patients. PMID- 19190869 TI - [Dementia--a secondary diagnosis in acute hospital care or more? Measures for improving care of hospitalized dementia patients]. AB - More and more people with dementia need acute care. However, a stay in the hospital involves severe risks for these patients: insufficient treatment, complications and a considerable emotional strain. A model project in Northrhine Westfalia provides acute care hospitals with measures to improve the care of patients with dementia. This article presents the conceptual results of the project referring to three fields: discharge and networking, medical treatment and general ward setting. Finally, the article points out some implications of the rising number of old age patients on health politics and research. PMID- 19190870 TI - [Gerontopsychiatric Consultation-Liaison Service--pioneers wait for imitators]. AB - The treatment and care of individuals with dementia, who are admitted to general hospitals due to somatic diseases, often leads to crisis situations which not only compromise the therapeutic benefit, but also can be followed by avoidable institutionalization in nursing homes or psychiatric hospitals. With the model project "Gerontopsychiatric Consultation-Liaison Service" in Kaufbeuren a comprehensive solution to this problem has been developed. A multiprofessional team consisting of a medical specialist, nurses specialized in psychiatry and an occupational therapist supports the health personnel by giving assistance and advice. It also provides contact persons for patients and their families. The effectiveness analysis proved that the work of this service has positive effects on both patient care and costs. Moreover a structural and process analysis provided data about the structure of patients and services as well as about the problems which arose during the setting up of the new service. Potential "imitators" can use this information in the implementation of similar services. PMID- 19190877 TI - Cardiac enzyme levels in myocardial dysfunction in newborns with perinatal asphyxia. AB - OBJECTIVE: To study the usefulness of cardiac enzymes in evaluating myocardial damage in perinatal asphyxia. METHODS: Thirty term babies with perinatal asphyxia and without any congenital malformations were selected as cases. They were compared with thirty healthy term babies without asphyxia. Myocardial dysfunction was evaluated using clinical, electrocardiography, echocardiography and cardiac enzymes i.e, troponin-T and CK-MB levels. RESULTS: Among the 30 cases 23 had evidence of myocardial involvement while one baby in the control group had ECG evidence compatible with cardiac involvement. Cardiac enzymes were significantly increased in babies with perinatal asphyxia. The mean level of C-troponin-T among cases and controls were 0.22+/-0.28 and 0.003+/-0.018 while CK-MB levels were 121+/-77.4 IU/L and 28.8 +/- 20.2 IU/L respectively. C-troponin-T had higher sensitivity and specificity compared to CK-MB levels. Moreover, C troponin-T levels correlated well with severity and outcome in babies with perinatal asphyxia. CONCLUSION: C-Tropopnin assay is useful in evaluating the severity of myocardial damage and outcome in perinatal asphyxia. PMID- 19190878 TI - Intravenous immune globulin versus intravenous anti-D immune globulin for the treatment of acute immune thrombocytopenic purpura. AB - OBJECTIVE: The purpose of this study was to compare the efficacy and side effects of intravenous immunoglobulin (IVIG) with intravenous anti-D immunoglobulin for treatment of newly diagnosed acute childhood Idiopathic thrombocytopenic purpura (ITP). METHODS: Children (6 months to 14 years) with newly diagnosed acute ITP and platelet count below 20,000/ microL were randomized to receive single dose intravenous 75 microg/kg anti-D or 1g/kg IVIG for two consecutive days (total dose 2 g/kg). Response rate defined as a platelet count over 20,000 / microL 72 hours after initial treatment. RESULTS: Eighty one patients (52 male and 29 female) with mean age of 5 years and 3 months randomly divided in anti-D group (n=42) and IVIG group (n=39). Mean baseline (pretreatment) platelet counts were 15406 / microL and 15230/ microL in anti-D and IVIG group, respectively. The response rate in IVIG group (98%) was more significant than anti-D group (76%); (P = 0.017). After 7 days the platelet counts of all patients in IVIG group were more than 20,000/ microL while in anti-D group 12% had platelet counts below 20,000/ microL. CONCLUSION: In acute childhood ITP, initial treatment with IVIG (2g/Kg in divided dose) increased platelet count more rapidly and more significant than intravenous anti-D (single dose of 75 microg/kg) within the first 72 hours. PMID- 19190879 TI - Association between bullying victimization and physical fighting among Filipino adolescents: results from the Global School-Based Health Survey. AB - OBJECTIVE: Interpersonal violence is an important, but neglected public health issue in low and middle-income countries. Adolescent physical fighting not only results in injury, disability and death, but is also associated with other potentially harmful behaviors such substance use and premarital sex. The study aims at dose-response association to prevent adolescent problem behaviors. METHODS: We used data from the 2003-2004 Global School-Based Health Survey conducted among school adolescents in the Philippines. We estimated the prevalence of bullying victimization and physical fighting. We also conducted logistic regression analysis to assess the association between a selected list of explanatory variables and physical fighting. We hypothesized that there would be a dose-response relationship between physical fighting and number of times the adolescent reported being bullied in the past 30 days. RESULTS: Of the 7,338 respondents, 35.5% (34.7% males and 36.1% females) were bullied and 50.0% (51.6% males and 48.8% females) reported having been in a physical fight in the past 12 months. There was a dose-response relationship between bullying victimization and physical fighting (p-trend <0.001). Compared to subjects who were not bullied, those who reported being bullied were more likely to engage in physical fighting after controlling for age, gender, substance use (smoking, alcohol drinking or drug use), and parental supervision (OR=2.38; 95% CI [1.99, 2.86] for 1-2 days of bullying victimization per month, OR=3.55; 95% CI [2.61, 4.83]) for 3-5 days/month per month, OR=4.45; 95% CI [2.61, 7.60]) for 6-9 days/ month, OR=1.91; 95% CI [1.17, 3.13]) for 10-30 days/month.) CONCLUSION: The dose-response relationship between physical fighting and the number of times an adolescent had been a victim of bullying deserves further study. If causal relationship exists, preventing bullying, even if not totally eliminated, may have significant results in preventing physical fighting. PMID- 19190880 TI - Cesarean section for suspected fetal distress, continuous fetal heart monitoring and decision to delivery time. AB - OBJECTIVE: To find out the efficacy of continuous fetal heart monitoring by analyzing the cases of cesarean section for nonreassuring fetal heart in labor, detected by cardiotocography (CTG) and correlating these cases with perinatal outcome. To evaluate whether a 30 minute decision to delivery (D-D) interval for emergency cesarean section influences perinatal outcome. METHODS: This was a prospective observational study of 217 patients who underwent cesarean section at > or = 36 weeks for non-reassuring fetal heart in labor detected by CTG. The maternal demographic profile, specific types of abnormal fetal heart rate tracing and the decision to delivery time interval were noted. The adverse immediate neonatal outcomes in terms of Apgar score <7 at 5 minutes, umbilical cord thornH <7.10, neonates requiring immediate ventilation and NICU admissions were recorded. The correlation between non-reassuring fetal heart, decision to delivery interval and neonatal outcome were analyzed. RESULTS: Out of 3148 patients delivered at > or = 36 weeks, 217 (6.8%) patients underwent cesarean section during labor primarily for non-reassuring fetal heart. The most common fetal heart abnormality was persistent bradycardia in 106 (48.8%) cases followed by late deceleration in 38 (17.5%) cases and decreased beat to beat variability in 17 (7.8%) cases. In 33 (15.2%) babies the 5 minutes Apgar score was <7 out of which 13 (5.9%) babies had cord thornH <7.10. Thirty three (15.2%) babies required NICU admission for suspected birth asphyxia. Rest 184 (84.7%) neonates were born healthy and cared for by mother. Regarding decision to delivery interval of < or =30 minutes versus >30 minutes, there was no significant difference in the incidence of Apgar score <7 at 5 minutes, cord pH <7.10 and new born babies requiring immediate ventilation. But the need for admission to NICU in the group of D-D interval < or = 30 minutes was significantly higher compared to the other group where D-D interval was >30 minutes. CONCLUSION: Non-reassuring fetal heart rate detected by CTG did not correlate well with adverse neonatal outcome. There was no significant difference in immediate adverse neonatal outcome whether the D-D time interval was < or = 30 minutes or >30 minutes; contrary to this, NICU admission for suspected birth asphyxia in 25 degrees), part-time bracing had to be stopped and changed to full-time bracing. We observed an overall success rate of 86.4% (patients with improved Cobb angles or halted progression). CONCLUSION: We were able to show a positive effect of part-time bracing in selected patients with mild scoliosis and a higher risk of progression. The negative medical and psychosocial consequences of 23-h brace treatment can therefore be avoided in certain patients. PMID- 19190892 TI - [Idiopathic scoliosis]. PMID- 19190890 TI - Increased insulin demand promotes while pioglitazone prevents pancreatic beta cell apoptosis in Wfs1 knockout mice. AB - AIMS/HYPOTHESIS: The WFS1 gene encodes an endoplasmic reticulum (ER) membrane embedded protein called Wolfram syndrome 1 protein, homozygous mutations of which cause selective beta cell loss in humans. The function(s) of this protein and the mechanism by which the mutations of this gene cause beta cell death are still not fully understood. We hypothesised that increased insulin demand as a result of obesity/insulin resistance causes ER stress in pancreatic beta cells, thereby promoting beta cell death. METHODS: We studied the effect of breeding Wfs1 ( -/- ) mice on a C57BL/6J background with mild obesity and insulin resistance, by introducing the agouti lethal yellow mutation (A ( y ) /a). We also treated the mice with pioglitazone. RESULTS: Wfs1 ( -/- ) mice bred on a C57BL/6J background rarely develop overt diabetes by 24 weeks of age, showing only mild beta cell loss. However, Wfs1 ( -/- ) A ( y ) /a mice developed selective beta cell loss and severe insulin-deficient diabetes as early as 8 weeks. This beta cell loss was due to apoptosis. In Wfs1 ( +/+ ) A ( y ) /a islets, levels of ER chaperone immunoglobulin-binding protein (BiP)/78 kDa glucose-regulated protein (GRP78) and phosphorylation of eukaryotic translation initiation factor 2, subunit alpha (eIF2alpha) apparently increased. Levels of both were further increased in Wfs1 ( -/- ) A ( y ) /a murine islets. Electron micrography revealed markedly dilated ERs in Wfs1 (-/-) A ( y ) /a murine beta cells. Interestingly, pioglitazone treatment protected beta cells from apoptosis and almost completely prevented diabetes development. CONCLUSIONS/INTERPRETATION: Wfs1-deficient beta cells are susceptible to ER stress. Increased insulin demand prompts apoptosis in such cells in vivo. Pioglitazone, remarkably, suppresses this process and prevents diabetes. As common WFS1 gene variants have recently been shown to confer a risk of type 2 diabetes, our findings may be relevant to the gradual but progressive loss of beta cells in type 2 diabetes. PMID- 19190895 TI - Comparison of two SPME fibers for the extraction of some off-flavor cork-taint compounds in bottled wines investigated by GC-HRMS. AB - Headspace solid-phase microextraction (HS-SPME) coupled to gas chromatography with high-resolution mass spectrometry (GC-HRMS) was used to determine 2,4,6 trichloroanisole, 2,3,6-trichloroanisole, 2,3,4-trichloroanisole, 2,3,5,6 tetrachloroanisole, pentachloroanisole, 2,4,6-tribromoanisole, 2 methylisoborneol, and 4-ethylguaiacol in wine samples. Two types of fiber coating commonly employed for sampling trichloroanisoles in wine and cork stoppers, viz. a polar mixed 50/30 microm divinylbenzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS) and a nonpolar 100 microm polydimethylsiloxane (PDMS), both 1 cm in length, were compared. This study demonstrates that the most employed polar mixed fiber should not be used with ethanol-water solutions, viz. wine samples, as its coating is not chemically and mechanically robust for sample extractions, as confirmed by environmental scanning electron microscopy. Much more effective and reliable results were obtained with the PDMS fiber, which remained functional for more than 80 analyses of red and white wine samples with satisfactory extraction efficiencies. Detection limits of investigated compounds, under optimized experimental conditions, ranged from 0.2 to 0.4 ng/L at a signal-to noise ratio of 3 and quantification limits from 0.8 to 1.5 ng/L. The proposed method was successfully applied to commercially available Italian white and red wines using 2,4,6-TCA-d(5) as the internal standard. PMID- 19190893 TI - Immediate postconditioning during reperfusion attenuates intestinal injury. AB - OBJECTIVE: To test the hypothesis that immediate but not delayed ischemic postconditioning (IPo) during reperfusion attenuates intestinal injury, and that ischemic preconditioning (IPC) and IPo may confer synergy in intestinal protection. DESIGN AND SETTING: Prospective laboratory animal study with concurrent control. SUBJECTS: Adult Sprague-Dawley rats. INTERVENTIONS: Intestinal ischemia/reperfusion (II/R) injury in rats was produced by clamping superior mesenteric artery for 60 min followed by 60 min reperfusion; IPC was elicited by 10 min ischemia and 10 min reperfusion before index ischemia; IPo was performed by three cycles of 30 s reperfusion and 30 s ischemia initiated either immediately at the onset of reperfusion (IPo) or after reperfusion for 3 min (delayed-IPo). Combination of IPC and IPo was performed by combining both protocols. MEASUREMENTS AND MAIN RESULTS: Intestinal ischemia/reperfusion resulted in significant intestinal injury evidenced as significant increase in Chiu's scores and wet-to-dry intestine weight ratio accompanied with increases in plasma levels of tumor necrosis factor-alpha and interleukin-6, as well as increases in the intestinal tissue lipid peroxidation product malonediadehyde and myeloperoxidase activity as compared to control animals (all P < 0.05). All these changes were significantly attenuated either by IPC or IPo or their combination (P < 0.05), and not by delayed-IPo (P > 0.05). IPC and IPo showed synergistic protection compared with either protocol alone. CONCLUSION: Ischemic postconditioning reduces intestinal injury, in part, by inhibiting oxidative injury, neutrophils filtration and proinflammatory response. The early period of reperfusion is critical to intestinal protection by IPo, and intestinal protection with IPo can be enhanced by IPC. PMID- 19190894 TI - Analysis of the mechanisms underlying the endothelium-dependent antivasoconstriction of puerarin in rat aorta. AB - Puerarin, a major isoflavonoid compound from the Chinese herb, Ge-gen (Pueraria lobata), has effective treatment on myocardial and cerebral ischemia, glaucoma and sudden deafness in clinical setting in China. Our present work showed that puerarin (50, 150, 450 microM) concentration-dependently inhibited phenylephrine or KCl-induced contraction only in endothelium-intact rat aortic rings. In Ca(2+) free solution, the antivasoconstriction of puerarin on phenylephrine was totally deprived. N (G)-nitro-L-arginine methyl ester, 1H-[1,2,4]oxadiazolo[4,3 a]quinoxalin-1-one, indomethacin and the three K(+) channel blockers, glibenclamide, tetraethylammonium and Ba(2+) displayed significant inhibitory effects on the antivasoconstriction of puerarin. 8-bromo-cGMP significantly strengthened the action of puerarin. Puerarin (10-160 microM) concentration dependently induced the NO production in the rat aortic cells. These findings suggested that the antivasoconstriction elicited by puerarin is endothelium dependent. NO/NO-cGMP pathway, PGI(2) and the opening of K(+) channels sensitive to glibenclamide, tetraethylammonium, and Ba(2+), which might be triggered by the extracellular Ca(2+) influx in the endothelium, appear to contribute to the antivasoconstriction of puerarin. PMID- 19190896 TI - Characterization of electrokinetic mobility of microparticles in order to improve dielectrophoretic concentration. AB - Insulator-based dielectrophoresis (iDEP), an efficient technique with great potential for miniaturization, has been successfully applied for the manipulation of a wide variety of bioparticles. When iDEP is applied employing direct current (DC) electric fields, other electrokinetic transport mechanisms are present: electrophoresis and electroosmotic flow. In order to concentrate particles, iDEP has to overcome electrokinetics. This study presents the characterization of electrokinetic flow under the operating conditions employed with iDEP; in order to identify the optimal conditions for particle concentration employing DC-iDEP, microparticle image velocimetry (microPIV) was employed to measure the velocity of 1-microm-diameter inert polystyrene particles suspended inside a microchannel made from glass. Experiments were carried out by varying the properties of the suspending medium (conductivity from 25 to 100 microS/cm and pH from 6 to 9) and the strength of the applied electric field (50-300 V/cm); the velocities values obtained ranged from 100 to 700 microm/s. These showed that higher conductivity and lower pH values for the suspending medium produced the lowest electrokinetic flow, improving iDEP concentration of particles, which decreases voltage requirements. These ideal conditions for iDEP trapping (pH = 6 and sigma(m) = 100 microS/cm) were tested experimentally and with the aid of mathematical modeling. The microPIV measurements allowed obtaining values for the electrokinetic mobilities of the particles and the zeta potential of the glass surface; these values were used with a mathematical model built with COMSOL Multiphysics software in order to predict the dielectrophoretic and electrokinetic forces exerted on the particles; the modeling results confirmed the microPIV findings. Experiments with iDEP were carried out employing the same microparticles and a glass microchannel that contained an array of cylindrical insulating structures. By applying DC electric fields across the insulating structures array, it was seen that the dielectrophoretic trapping was improved when the electrokinetic force was the lowest; as predicted by microPIV measurements and the mathematical model. The results of this study provide guidelines for the selection of optimal operating conditions for improving insulator-based dielectrophoretic separations and have the potential to be extended to bioparticle applications. PMID- 19190897 TI - Critical evaluation of novel dynamic flow-through methods for automatic sequential BCR extraction of trace metals in fly ash. AB - Two novel dynamic extraction approaches, the so-called sequential injection microcolumn extraction and sequential injection stirred-flow chamber extraction, based on the implementation of a sample-containing container as an external extraction reactor in a sequential injection network, are for the first time, optimized and critically appraised for fractionation assays. The three steps of the original Community Bureau of Reference (BCR) sequential extraction scheme have been performed in both automated dynamic fractionation systems to evaluate the extractability of Cr, Cu, Ni, Pb, and Zn in a standard reference material of coal fly ash (NIST 1633b). In order to find the experimental conditions with the greatest influence on metal leachability in dynamic BCR fractionation, a full factorial design was applied, in which the solid sample weight (100-500 mg) and the extraction flow rate (3.0-6.0 mL min(-1)) were selected as experimental factors. Identical cumulative extractabilities were found in both sequential injection (SI)-based methods for most of assayed trace elements regardless of the extraction conditions selected, revealing that both dynamic fractionation systems, as opposed to conventional steady-state BCR extraction, are not operationally defined within the selected range of experimental conditions. Besides, the proposed automated SI assemblies offer a significant saving of operational time with respect to classical BCR test, that is, 3.3 h versus 48 h, for complete fractionation with minimum analyst involvement. PMID- 19190898 TI - Oblique meniscomeniscal ligament: a normal variant. PMID- 19190899 TI - Autoimmune pancreatitis in an 11-year-old boy. AB - We report a case of histopathologically proven autoimmune pancreatitis in an 11 year-old boy. Abdominal US and MRI showed a focal swelling of the pancreatic head, the latter also showing delayed contrast enhancement. There was diffuse irregular pancreatic duct narrowing, compression of the intrapancreatic common bile duct, and mild proximal biliary dilatation on MR cholangiopancreatography. Laboratory results revealed normal serum IgG and subclass 4 with negative autoimmune antibodies, and slightly elevated carbohydrate antigen 19-9. This highlights the differentiation of autoimmune pancreatitis from pancreatic head cancer and, to a lesser extent, other forms of pancreatitis in children. PMID- 19190902 TI - A novel family VIII carboxylesterase derived from a leachate metagenome library exhibits promiscuous beta-lactamase activity on nitrocefin. AB - The realization that majority of microbes are not amenable to cultivation as isolates under laboratory conditions has led to the culture-independent metagenomic approach as a novel technique for novel biocatalyst discovery. A leachate fosmid shotgun metagenome library was constructed and subsequently screened for esterolytic activities on a tributyrin agar medium. Nucleotide sequencing and translational analysis of an esterase-positive fosmid clone led to the identification of a 1,281 bp esterase gene (estC) encoding a protein (EstC) of 427 aa with translated molecular weight of 46.3 kDa. The EstC primary structure contained a signal leader peptide (29 aa), which could be cleaved to form a mature protein of 398 aa with molecular weight 43.3 kDa. Homology searches revealed that EstC belonged to the family VIII esterases, which exploit a serine residue within the S-x-x-K motif as a catalytic nucleophile. Substrate specificity studies showed that EstC prefers short to medium acyl chain length of p-nitrophenyl esters, a characteristic typical of "true" carboxylesterases. Moreover, EstC represents the first member of the family VIII esterases with a leader peptide and a detectable promiscuous beta-lactam hydrolytic activity. Site directed mutagenesis studies also revealed that in addition to Ser103 and Lys106 residues, the Tyr219 residue also plays a catalytic role in EstC. The organic solvent stability and the specificity towards esters of tertiary alcohols linalyl acetate (3,7-dimethyl-1,6-octadien-3-yl acetate) make EstC potentially useful in biocatalysis. PMID- 19190903 TI - Optimization and production of novel antimicrobial agents from sponge associated marine actinomycetes Nocardiopsis dassonvillei MAD08. AB - The sponge-associated actinomycetes were isolated from the marine sponge Dendrilla nigra, collected from the southwest coast of India. Eleven actinomycetes were isolated depending upon the heterogeneity and stability in subculturing. Among these, Nocardiopsis dassonvillei MAD08 showed 100% activity against the multidrug resistant pathogens tested. The culture conditions of N. dassonvillei MAD08 was optimized under submerged fermentation conditions for enhanced antimicrobial production. The unique feature of MAD08 includes extracellular amylase, cellulase, lipase, and protease production. These enzymes ultimately increase the scope of optimization using broad range of raw materials which might be efficiently utilized. The extraction of the cell free supernatant with ethyl acetate yielded bioactive crude extract that displayed activity against a panel of pathogens tested. Analysis of the active thin layer chromatography fraction by Fourier transform infrared and gas chromatography-mass spectrometry evidenced 11 compounds with antimicrobial activity. The ammonium sulfate precipitation of the culture supernatant at 80% saturation yielded an anticandidal protein of molecular weight 87.12 kDa. This is the first strain that produces both organic solvent and water soluble antimicrobial compounds. The active extract was non-hemolytic and showed surface active property envisaging its probable role in inhibiting the attachment of pathogens to host tissues, thus, blocking host-pathogen interaction at an earlier stage of pathogenesis. PMID- 19190904 TI - Mutations at subsite -3 in cyclodextrin glycosyltransferase from Paenibacillus macerans enhancing alpha-cyclodextrin specificity. AB - A major disadvantage of cyclodextrin production is the limited cyclodextrin product specificity of cyclodextrin glycosyltransferase (CGTase). Here, we described mutations of Asp372 and Tyr89 at subsite -3 in the CGTase from Paenibacillus macerans strain JFB05-01. The results showed that Asp372 and Tyr89 played important roles in cyclodextrin product specificity of CGTase. The replacement of Asp372 by lysine and Tyr89 by aspartic acid, asparagine, lysine, and arginine resulted in a shift in specificity towards the production of alpha cyclodextrin, which was most apparent for the mutants D372K and Y89R. Furthermore, the changes in cyclodextrin product specificity for the single mutants D372K and Y89R could be combined in the double mutant D372K/Y89R, which displayed a 1.5-fold increase in the production of alpha-cyclodextrin, with a concomitant 43% decrease in the production of beta-cyclodextrin when compared to the wild-type CGTase. Thus, the D372K and Y89R single and double mutants were much more suitable for the industrial production of alpha-cyclodextrin than the wild-type enzyme. The enhanced alpha-cyclodextrin specificity of these mutants might be a result of stabilizing the bent conformation of the intermediate in the cyclization reaction. PMID- 19190906 TI - Lung ventilation-perfusion scintigraphy in children on long-term parenteral nutrition. AB - PURPOSE: Pulmonary emboli (PE) are one of the major complications associated with total parenteral nutrition (TPN). Ventilation-perfusion scintigraphy (V/Q) remains the most used test for the diagnosis of PE and follow-up of patients on TPN. The aim of our study was to demonstrate the high prevalence of undiagnosed PE in children on TPN. METHODS: The medical and imaging files of 64 patients on TPN who underwent V/Q examinations covering the period of 1986-2004 were reviewed. Children were aged between 3.18 months and 21.6 years. TPN was started at birth (range 0-15 years). All children had a normal chest radiograph and no symptoms at the time of the V/Q scan. A comparative analysis between the prevalence of PE and risk factors (number of days per week with lipophilic content of the TPN, bowel inflammation and thrombophilic factors (protein C and S) was performed. RESULTS: Of the 64 patients, 25 (39%) had an abnormal V/Q scan. A total of 29 PE episodes were diagnosed in all patients. Two children had three episodes of PE. The median age at PE diagnosis was 4.6 years. In 17 patients (68%) diagnosis was achieved on the first V/Q scan performed. PE was bilateral in 56% and unilateral in 44%. PE was the main cause of 2 out 15 recorded deaths. All risk factors were associated with an increase in PE prevalence by statistical analysis. CONCLUSION: PE is underdiagnosed in children on long-term TPN. Lung V/Q scintigraphy is useful in the diagnosis of PE in children with a low pretest probability. PMID- 19190907 TI - Synthesis of sialyl Lewis(a) (sLe (a), CA19-9) and construction of an immunogenic sLe(a) vaccine. AB - Sialyl Lewis(a) (sLe(a)), also termed CA19-9 antigen, is recognized by murine mAb19-9 and is expressed on the cancer cell surface as a glycolipid and as an O linked glycoprotein. It is highly expressed in a variety of gastrointestinal epithelial malignancies including colon cancer and pancreatic cancer, and in breast cancer and small cell lung cancer, but has a limited expression on normal tissues. sLe(a) is known to be the ligand for endothelial cell selectins suggesting a role for sLe(a) in cancer metastases and adhesion. For these reasons, sLe(a) may be a good target for antibody mediated immunotherapy including monoclonal antibodies and tumor vaccines. However, sLe(a) is structurally similar to sLe(x) and other blood group related carbohydrates which are widely expressed on polymorphonucleocytes and other circulating cells, raising concern that immunization against sLe(a) will induce antibodies reactive with these more widely expressed autoantigens. We have shown previously both in mice and in patients that conjugation of a variety of carbohydrate cancer antigen to keyhole limpet hemocyanin (KLH) and administration of this conjugate mixed with saponin adjuvants QS-21 or GPI-0100 are the most effective methods for induction of antibodies against these cancer antigens. We describe here for the first time the total synthesis of pentenyl glycoside of sLe(a) hexasaccharide and its conjugation to KLH to construct a sLe(a)-KLH conjugate. Groups of five mice were vaccinated subcutaneously four times over 6 weeks. Sera were tested against sLe(a)-HSA by ELISA and against sLe(a) positive human cell lines adenocarcinoma SW626 and small cell lung cancer (SCLC) DMS79 by FACS. As expected, mice immunized with unconjugated sLe(a) plus GPI-0100 or unconjugated sLe(a) mixed with KLH plus GPI-0100 failed to produce antibodies against sLe(a). However, mice immunized with sLe(a)-KLH conjugate without GPI-0100 produced low levels of antibodies and mice immunized with sLe(a)-KLH plus GPI-0100 produced significantly higher titer IgG and IgM antibodies against sLe(a) by ELISA. These antibodies were highly reactive by FACS and mediated potent complement mediated cytotoxicity against sLe(a) positive SW626 and DMS79 cells. They showed no detectable cross reactivity against a series of other blood group-related antigens, including Le(y), Le(x), and sLe(x) by dot blot immune staining. This vaccine is ready for testing as an active immunotherapy for treating sLe(a) positive cancer in clinical settings. PMID- 19190908 TI - Dose-finding study of docetaxel, oxaliplatin, and S-1 for patients with advanced gastric cancer. AB - PURPOSE: To determine the maximum tolerated dose (MTD), recommended dose (RD), and activity of combined docetaxel, oxaliplatin, and S-1 (DOS) chemotherapy on metastatic gastric cancer. PATIENTS AND METHODS: Docetaxel and oxaliplatin were administered intravenously on day 1 and S-1 was administered orally on days 1-14 of every 21-day cycle. The doses of docetaxel/oxaliplatin/S-1 in the phase I study were level -1A, 52.5/80/60 mg/m(2); level -1B, 52.5/80/80 mg/m(2); level 1A, 52.5/105/80 mg/m(2); level 1B, 52.5/130/80 mg/m(2); level 2A, 60/105/80 mg/m(2); level 2B, 60/130/80 mg/m(2); level 3A, 67.5/105/80 mg/m(2); level 3B, 67.5/130/80 mg/m(2); level 4A, 75/105/80 mg/m(2); level 4B, 75/130/80 mg/m(2). RESULTS: Nine patients were enrolled. One of six patients at level 1A and two of three patients at level 1B developed dose-limiting toxicity (febrile neutropenia) during the initial two cycles. Therefore, the doses used at levels 1B and 1A were defined as the MTD and RD, respectively. All patients were evaluated for toxicity and response. Six partial responses were noted, and the overall response rate was 67%. CONCLUSION: The RD of the DOS regimen in patients with advanced gastric cancer was docetaxel 52.5 mg/m(2) and oxaliplatin 105 mg/m(2) on day 1 and S-1 80 mg/m(2) on days 1-14 of every 21-day cycle. A phase II study using the RD is currently underway. PMID- 19190909 TI - [New aspects in the pathology of the mammary gland]. PMID- 19190910 TI - High b-value diffusion-weighted MRI for detecting gallbladder carcinoma: preliminary study and results. AB - The aim of this preliminary study was to retrospectively evaluate the usefulness of high b-value diffusion-weighted MR imaging (DWI) in the detection of gallbladder carcinoma. Fifteen patients with gallbladder carcinoma and 14 other patients were included in this study. All patients and subjects underwent DWI, and images were evaluated by two radiologists. The area under the receiver operating characteristic curve (AUC), apparent diffusion coefficient (ADC) measurement, sensitivity and specificity were calculated. An AUC yielded 0.980 (95% CI, 0.850-0.999) and 0.941 (95% CI, 0.791-0.990) for the two radiologists. The mean sensitivity and specificity were 83.3% and 100%, respectively. The mean ADC value of gallbladder carcinoma was (1.28 +/- 0.41)x10(-3) mm(2)/s and that of control gallbladder lesions was (1.92 +/- 0.21)x10(-3) mm(2)/s (P < 0.01). According to the results of our preliminary study, high b-value DWI might be a useful tool for detecting gallbladder carcinoma by measuring the ADC value and direct visual assessment. PMID- 19190911 TI - Radiation dose of cardiac CT--what is the evidence? AB - Current evidence and most pertinent literature on the radiation dose of cardiac computed tomography (CT) for the noninvasive assessment of coronary artery disease are reviewed. The various means for adjusting CT protocols to lower the radiation to a level that is as low as reasonably achievable are discussed. It is shown that for the target population of cardiac CT, the direct visualization of the heart and coronary arteries outweighs the hypothetical risk of the investigation, provided that indications are prudent and the protocols appropriate. PMID- 19190912 TI - Percutaneous treatment of intrabdominal abscess: urokinase versus saline serum in 100 cases using two surgical scoring systems in a randomized trial. AB - The purpose of this study was to assess whether regular instillation of urokinase during abscess drainage leads to an improved outcome compared to saline irrigation alone. One hundred patients referred for image-guided abdominal abscess drainage were randomized between thrice daily urokinase instillation or saline irrigation alone. At the end of the study, patient medical records were reviewed to determine drainage, study group, Altona (PIA II) and Mannheim (MPI) scoring, duration of drainage, procedure-related complications, hospital stay duration, and clinical outcome. The technical success rate of the percutaneous abscess drainage was 100%. The success or failure of abscess remission did not differ significantly between groups (success rate of 91.5% in the urokinase group vs. 88.8% in the saline group; failure rate was of 8.5 vs. 21.2%, respectively); however, days of drainage, main hospital stay, and overall costs were significantly reduced in patients treated with urokinase compared to the control group (P < 0.05). No adverse effects from urokinase were observed. Surgical scores were a useful homogeneity factor, and MPI showed a good correlation with prognosis, while PIA results did not have a significant correlation. For drainage of complex abscesses (loculations, hemorrhage, viscous material), fibrinolytics safely accelerate drainage and recovery, reducing the length of the hospital stay and, therefore, the total cost. PMID- 19190913 TI - Mushroom-cage gastrostomy tube placement in patients with amyotrophic lateral sclerosis: a 5-year experience in 104 patients in a single institution. AB - To review our experience of placement of a mushroom-cage gastrostomy tube (Entristar, Tyco Healthcare, Mansfield, MA), using radiological guidance, in patients with amyotrophic lateral sclerosis (ALS). All procedures were performed under local anaesthesia without sedation. Complications were recorded as peri procedural, early (<24 h), late (>24 h), major or minor. Deaths were recorded as related to the underlying ALS or secondary to radiological-inserted gastrostomy (RIG) placement. Replacement RIG tube rate was recorded. Over a 5-year period RIG tubes were placed in 104 patients with ALS (male n = 52, female n = 52), with a median age of 62 years (range 34-86 years). All procedures were technically successful. Of the RIG procedures, 21/104 (20.2%) were performed with respiratory support. The 30-day mortality rate was 7/104 (6.7%); no patient died as a result of the procedure. There were 23/104 (22.1%) complications overall; 20/104 (19.2%) were minor and 3/104 (2.9%) major, requiring surgery (n = 2) and radiological guided abscess drainage (n = 1). A median interval between replacement RIG procedures in 20/104 (19.2%) patients was 141.5 days (range 43-537 days). A mushroom-cage RIG tube may be safely and effectively inserted in a 'one-step' radiological procedure and may replace endoscopic-inserted gastrostomy tubes in the nutritional management of ALS. PMID- 19190914 TI - Assessment of the spatial pattern of colorectal tumour perfusion estimated at perfusion CT using two-dimensional fractal analysis. AB - The aim was to evaluate the feasibility of fractal analysis for assessing the spatial pattern of colorectal tumour perfusion at dynamic contrast-enhanced CT (perfusion CT). Twenty patients with colorectal adenocarcinoma underwent a 65-s perfusion CT study from which a perfusion parametric map was generated using validated commercial software. The tumour was identified by an experienced radiologist, segmented via thresholding and fractal analysis applied using in house software: fractal dimension, abundance and lacunarity were assessed for the entire outlined tumour and for selected representative areas within the tumour of low and high perfusion. Comparison was made with ten patients with normal colons, processed in a similar manner, using two-way mixed analysis of variance with statistical significance at the 5% level. Fractal values were higher in cancer than normal colon (p < or = 0.001): mean (SD) 1.71 (0.07) versus 1.61 (0.07) for fractal dimension and 7.82 (0.62) and 6.89 (0.47) for fractal abundance. Fractal values were lower in 'high' than 'low' perfusion areas. Lacunarity curves were shifted to the right for cancer compared with normal colon. In conclusion, colorectal cancer mapped by perfusion CT demonstrates fractal properties. Fractal analysis is feasible, potentially providing a quantitative measure of the spatial pattern of tumour perfusion. PMID- 19190915 TI - Remodelling of the aortic root in severe tricuspid aortic stenosis: implications for transcatheter aortic valve implantation. AB - Detailed knowledge of aortic root geometry is a prerequisite to anticipate complications of transcatheter aortic valve (TAV) implantation. We determined coronary ostial locations and aortic root dimensions in patients with aortic stenosis (AS) and compared these values with normal subjects using computed tomography (CT). One hundred consecutive patients with severe tricuspid AS and 100 consecutive patients without valvular pathology (referred to as the controls) undergoing cardiac dual-source CT were included. Distances from the aortic annulus (AA) to the left coronary ostium (LCO), right coronary ostium (RCO), the height of the left coronary sinus (HLS), right coronary sinus (HRS), and aortic root dimensions [diameters of AA, sinus of Valsalva (SV), and sino-tubular junction (STJ)] were measured. LCO and RCO were 14.9 +/- 3.2 mm (8.2-25.9) and 16.8 +/- 3.6 mm (12.0-25.7) in the controls, 15.5 +/- 2.9 mm (8.8-24.3) and 17.3 +/- 3.6 mm (7.3-26.0) in patients with AS. Controls and patients with AS had similar values for LCO (P = 0.18), RCO (P = 0.33) and HLS (P = 0.88), whereas HRS (P < 0.05) was significantly larger in patients with AS. AA (r = 0.55,P < 0.001), SV (r = 0.54,P < 0.001), and STJ (r = 0.52,P < 0.001) significantly correlated with the body surface area in the controls; whereas no correlation was found in patients with AS. Patients with AS had significantly larger AA (P < 0.01) and STJ (P < 0.01) diameters when compared with the controls. In patients with severe tricuspid AS, coronary ostial locations were similar to the controls, but a transverse remodelling of the aortic root was recognized. Owing to the large distribution of ostial locations and the dilatation of the aortic root, CT is recommended before TAV implantation in each patient. PMID- 19190916 TI - Aperistaltic effect of hyoscine N-butylbromide versus glucagon on the small bowel assessed by magnetic resonance imaging. AB - The aim of this prospective study was to compare the intraindividual aperistaltic effect of 40 mg hyoscine N-butylbromide (HBB/Buscopan) with that of 1 mg glucagon on small bowel motility by using magnetic resonance imaging (MRI). Ten healthy volunteers underwent two separate 1.5-T MRI studies (HBB/glucagon) after a standardized oral preparation with an aqueous solution of Gd-DOTA and ispaghula (Metamucil). A 2D T1-w GRE sequence was acquired (TR 2.7 ms/TE 1.3 ms, temporal resolution 0.25 s) before and after intravenous (i.v.) drug administration and motility was followed over 1 h. On the resulting images the cross-sectional luminal diameters were assessed and plotted over time. Baseline motility frequency, onset of aperistalsis, duration of arrest, reappearance of motility and return to normal motility were analysed. Significant differences regarding reliability and duration of aperistalsis were observed. In the HBB group aperistalsis lasted a mean of 6.8 +/- 5.3 min compared with 18.3 +/- 7 min after glucagon (p < 0.0001). In 50% of cases HBB did not accomplish aperistalsis, whereas glucagon always succeeded (p = 0.05). There were no significant differences in terms of baseline and end frequencies for the onset of aperistalsis (22.2 +/- 37.5 s HBB/13.4 +/- 9.2 s glucagon, p = 0.1), nor for the return to normal motility. Arrest of small bowel motion is achieved more reliably and lasts significantly longer after i.v. administration of 1 mg glucagon compared with 40 mg HBB. PMID- 19190918 TI - Correlation of in vivo bladder blood flow measurements with tissue hypoxia. AB - AIMS: Obstructive bladder dysfunction is in part due to reduced blood flow and the resulting ischemia of the bladder smooth muscle and mucosa. Our aim was to determine if the severity and localization of ischemia could be determined by measuring blood flow to the bladder with a non-invasive probe placed on the surface of the urothelium. MATERIALS AND METHODS: Twenty-four adult male rabbits (5 months, 3.5-4.0 kg) were divided into three groups: 1-controls; 2-2 h of bilateral ischemia; and 3-partial outlet obstruction, and were evaluated after 2 weeks. Each rabbit received an intraperitoneal injection of Hypoxyprobe-1. In vivo real-time monitoring of blood flow was measured at five sites within the bladders with a laser Doppler flowmeter. RESULTS: For all groups, the blood flow readings showed no significant differences among the five sites. The ischemic bladders showed significant decreases in blood flow. The obstructed bladders had significantly lower blood flow than the ischemic bladders. The hypoxyprobe studies demonstrated that there was no hypoxia present in the control bladders; the mucosa of the ischemic bladders showed even hypoxia at an intermediate concentration; the obstructed bladders showed dense but even staining. CONCLUSION: We have demonstrated that we can determine the severity of ischemia by surface measurement of blood flow. PMID- 19190917 TI - Whole-body MRI at 1.5 T and 3 T compared with FDG-PET-CT for the detection of tumour recurrence in patients with colorectal cancer. AB - The purpose of this study was to assess the diagnostic accuracy of whole-body MRI (WB-MRI) at 1.5 T or 3 T compared with FDG-PET-CT in the follow-up of patients suffering from colorectal cancer. In a retrospective study, 24 patients with a history of colorectal cancer and suspected tumour recurrence underwent FDG-PET-CT and WB-MRI with the use of parallel imaging (PAT) for follow-up. High resolution coronal T1w-TSE and STIR sequences at four body levels, HASTE imaging of the lungs, contrast-enhanced T1w- and T2w-TSE sequences of the liver, brain, abdomen and pelvis were performed, using WB-MRI at either 1.5 T (n = 14) or 3 T (n = 10). Presence of local recurrent tumour, lymph node involvement and distant metastatic disease was confirmed using radiological follow-up within at least 5 months as a standard of reference. Seventy seven malignant foci in 17 of 24 patients (71%) were detected with both WB-MRI and PET-CT. Both investigations concordantly revealed two local recurrent tumours. PET-CT detected significantly more lymph node metastases (sensitivity 93%, n = 27/29) than WB-MRI (sensitivity 63%, n = 18/29). PET-CT and WB-MRI achieved a similar sensitivity for the detection of organ metastases with 80% and 78%, respectively (37/46 and 36/46). WB-MRI detected brain metastases in one patient. One false-positive local tumour recurrence was indicated by PET-CT. Overall diagnostic accuracy for PET-CT was 91% (sensitivity 86%, specificity 96%) and 83% for WB-MRI (sensitivity 72%, specificity 93%), respectively. Examination time for WB-MRI at 1.5 T and 3 T was 52 min and 43 min, respectively; examination time for PET-CT was 103 min. Initial results suggest that differences in accuracy for local and distant metastases detection using FDG-PET-CT and WB-MRI for integrated screening of tumour recurrence in colorectal cancer depend on the location of the malignant focus. Our results show that nodal disease is better detected using PET-CT, whereas organ disease is depicted equally well by both investigations. PMID- 19190919 TI - [Clinical findings and diagnostics of cone dystrophy]. AB - BACKGROUND: Cone dystrophies present with highly variable clinical findings and often limited retinal changes, which may lead to misdiagnosis. The purpose of the present review of the clinical presentation and diagnosis of cone dystrophies is to provide guidelines for improved patient care. METHODS: A literature search and evaluation of the clinical findings were carried out in 450 patients with cone dystrophy examined between 1986 and 2008. RESULTS: Characteristic signs are loss of visual acuity, photophobia and central scotoma. The diagnosis of cone dystrophy is determined by a full-field electroretinogram (ERG). Fundus and near infrared autofluorescence as well as optical coherence tomography allow detection of retinal structural abnormalities even when findings from ophthalmoscopy are normal. CONCLUSION: The diagnosis of cone dystrophy is difficult due to unspecific subjective symptoms and absence of characteristic ophthalmoscopic findings. The differential diagnosis of unexplained visual loss should include cone dystrophy and requires either a full-field or multifocal ERG. PMID- 19190920 TI - Evolution of photoperiodic time measurement is independent of the circadian clock in the pitcher-plant mosquito, Wyeomyia smithii. AB - For over 70 years, researchers have debated whether the ability to use day length as a cue for the timing of seasonal events (photoperiodism) is related to the endogenous circadian clock that regulates the timing of daily events. Models of photoperiodism include two components: (1) a photoperiodic timer that measures the length of the day, and (2) a photoperiodic counter that elicits the downstream photoperiodic response after a threshold number of days has been counted. Herein, we show that there is no geographical pattern of genetic association between the expression of the circadian clock and the photoperiodic timer or counter. We conclude that the photoperiodic timer and counter have evolved independently of the circadian clock in the pitcher-plant mosquito Wyeomyia smithii and hence, the evolutionary modification of photoperiodism throughout the range of W. smithii has not been causally mediated by a corresponding evolution of the circadian clock. PMID- 19190922 TI - Double rectal perforation after stapled haemorrhoidectomy. PMID- 19190921 TI - Surgical treatment of sigmoid diverticulitis--analysis of predictive risk factors for postoperative infections, surgical complications, and mortality. AB - BACKGROUND AND AIMS: Sigmoid diverticular disease has great clinical importance due to its increasing incidence in the Western world and a broad spectrum of clinical features with potential fatal complications after surgery. The definition of risk factors associated with postoperative infections, surgical complications and mortality could be helpful in clinical decision-making and optimizing perioperative treatment. MATERIALS AND METHODS: Based on a prospective database, 168 consecutive patients undergoing surgery for sigmoid diverticulitis were included in this study. The association of different potential risk factors such as age, Hinchey classification, type and duration of operation, surgeons' experience, blood loss, comorbidities, and hospital course with perioperative complications and mortality were tested by univariate and multivariate analysis. RESULTS: Of the 168 patients enrolled in this study, there were 84 male and 84 female. A third of patients were operated as emergency cases (within 24 h after surgical evaluation); 62% underwent open surgery, 35% were treated laparoscopically with a conversion rate of 3%. A blood transfusion received 14% of patients, a surgical infection occurred in 20%, surgical complications appeared in 24% with a necessity for re-exploration in 9.5%. Leakage of the primary anastomosis was seen in 3.3%, whereas a leakage of the Hartmann's stump occurred in 4.3%. Overall in-hospital mortality was 4.1%. Multivariate analysis demonstrated Hinchey classification and intraoperative blood transfusion to be independently associated with postoperative infections, complications and mortality. CONCLUSION: Hinchey classification and intraoperative blood transfusion are independently associated with a worse perioperative outcome in patients undergoing surgery for sigmoid diverticular disease. While Hinchey classification cannot be influenced per se by the surgeon, outcome might be influenced by reducing the need for intraoperative blood transfusion. PMID- 19190923 TI - Truncated tau at D421 is associated with neurodegeneration and tangle formation in the brain of Alzheimer transgenic models. AB - In addition to tau hyperphosphorylation, tau truncation is also detected in Alzheimer's disease (AD) patients. In the brain of AD transgenic mouse models, the pathological details of truncated tau are not well characterized. In this study, we analyzed spatial relationships among tau truncation, tau phosphorylation and neurodegeneration or tangle formation in a tau(P301L) single transgenic mouse model and a triple transgenic mouse model that produces both amyloid plaques, and neurofibrillary tangles. During development of tau pathology, the spatial relationship between hyperphosphorylation and truncation of tau exhibited a shift from colocalization at low densities of hyperphosphorylated tau to partial dissociation at high densities. Importantly, we detected a few neurons that contained abundant truncated tau but were lacking hyperphosphorylation, and these neurons exhibited remarkable nuclear condensation. In the case of colocalization, truncated tau was commonly associated with high immunoreactivity of hyperphosphorylated tau and dense Gallyas silver staining. Taken together, our study shows that tau truncation appears after tau hyperphosphorylation in the brain of two transgenic mouse models, and that accumulation of truncated tau, in the absence or the presence of phosphorylated tau, is closely associated with a subset of neurons undergoing degeneration or containing neurofibrillary tangles. PMID- 19190924 TI - A new distractable implant for vertebral body replacement: biomechanical testing of four implants for the thoracolumbar spine. AB - INTRODUCTION: Expandable titanium implants for vertebral body replacement in the thoracolumbar spine have been well established in the reconstruction of the anterior spinal column. Load transfer at the bone-implant interface remains a point of concern. The purpose of the study was to compare the performance in axial load transfer from the implant to the vertebral body in four different implants, all of them in clinical use to date. MATERIALS AND METHODS: We tested a second generation implant (Synex II) in comparison to three different expandable titanium cages: Synex I, Obelisc and X-Tenz. Twenty-four intact fresh frozen human lumbar vertebrae (L1-L4) were distributed into four identical groups according to bone mineral density (BMD). The BMD was determined by quantitative computed tomography (qCT). Specimens were loaded in craniocaudal direction with a material testing machine (Mini Bionix II) at a constant speed of 5 mm/min. Load displacement curves were continuously recorded for each specimen until failure (diminishment of compressive force (F) and/or obvious implant migration through the vertebral body end plate). One-way analysis of variance (ANOVA) and post-hoc tests (Bonferroni) were applied to detect differences at 1, 2, 3, and 4 mm displacement (F (1-4 mm)) between implant groups. RESULT: No significant differences were observed with regard to maximum compression force (F (max)) and displacement (d (max)) until failure: Synex II (1,782.3 N/4.67 mm); Synex I (1,645.3 N/4.72 mm); Obelisc (1,314.0 N/4.24 mm); X-Tenz (1470.3 N/6.92 mm). However, the mean compression force at 1-4 mm displacement (F (1-4 mm): 300-1,600 N) was highest for Synex II. The difference at 2 mm displacement was significant (p = 0.028) between Synex II (F (2 mm) = 879 N) and X-Tenz (F (2 mm) = 339 N). CONCLUSION: The modified end plate design of Synex II was found to perform comparably at least with regard to the compressive performance at the implant bone interface. The risk of the new implant for collapse into the vertebral body might be reduced when compared to the competitors. PMID- 19190925 TI - Worldwide first successful splitting of the breast for a single-procedure reconstruction after mastectomy with maintaining the sensitivity. AB - PURPOSE: Breast reconstructions are made autologously, that is with the body's own tissue, or heterologously by means of implants. This either causes donor site morbidity or a foreign body is implanted. Both are disadvantages which could not be avoided up to now. By using tissue of the still remaining (contralateral) breast both could be avoided. The object of this paper is to check whether this technique is feasible. METHOD: By dividing the existing breast and transferring it to the contralateral side, we were able to successfully conduct a single procedure breast reconstruction with one female patient. RESULTS: The operation technique as well as the post-operative progression of the female patient will be presented. CONCLUSION: Mamma-splitting is a new and promising method of reconstruction with own tissue, without donor site morbidity or implant. The use within a bigger group of female patients will show the method's validity. PMID- 19190926 TI - A new technique for the diagnosis of fallopian tube patency by using hysteroscopy with ultrasound compared with hysterosalpingography in infertile women. AB - PURPOSE: To evaluate the effectiveness of hysteroscopy as a method for the diagnosis of tubal patency using saline distention media. METHODS: In this prospective study, 62 women between 21 and 38 years with a history of at least 1 year of infertility who underwent hysteroscopy and hysterosalpingography (HSG) on two consecutive cycles were studied. Transvaginal ultrasonography carried out before and after hysteroscopy in order to measure the abdominal fluid (in ml) in the peritoneal cavity of the cul-de-sac. The difference between the two results was calculated. A next cycle HSG was performed and patients were divided into three groups according to whether there was no tubal occlusion (group 1), a unilateral occlusion (group 2) or a bilateral occlusion (group 3): Peritoneal fluid measurements were compared among the three groups. Finally, pain and discomfort were recorded at the end of the process. RESULTS: According to the HSG, 34 women were in group 1, 13 in group 2 and 9 in group 3. Peritoneal fluid measurements were 6.88 +/- 2.7 ml in group 1, 4.21 +/- 0.9 ml in group 2 and 1.08 +/- 0.7 ml in group 3. Statistical differences were found between groups 1 and 3 and between groups 2 and 3 (P < 0.05). All of the patients reported significantly less pain during hysteroscopy in response to HSG. CONCLUSIONS: Using saline distension media during hysteroscopic evaluation was effective to allow measurement of the accumulating fluid in the peritoneal cavity and to confirm at least one patent tube with minimal pain. PMID- 19190927 TI - The efficacy of Implanon for the treatment of chronic pelvic pain associated with pelvic congestion: 1-year randomized controlled pilot study. AB - OBJECTIVE: To evaluate the beneficial effects of Implanon on pelvic pain in women with pelvic congestion syndrome (PCS). The efficacy of pain control, amount and frequency of menstrual loss, degree of patient's satisfaction and objective pelvic venography scores were investigated. METHODS: In a prospective open labelled study, 25 consecutive women complaining of chronic pelvic pain were recruited. Pretreatment objective peruterine venography and diagnostic laparoscopy of pure PCS together with subjective pelvic pain scores, prefilled questionnaire of Hospital Anxiety and Depression Scale (HADS), visual analogue scale (VAS), verbal rating scale (VRS) and quantified menstrual loss using the pictorial blood loss chart were documented in all cases. After identification, 23 subjects with pure PCS were randomly assigned to have either Implanon inserted subcutaneously (12 cases) or no treatment (11 cases). Patients were followed up at 1, 3, 6, 9 and 12 months. A symptom diary for side effects, VAS, VRS and menstrual scores were used to assess the subjective response to treatment. At the end of the study, all patients underwent repeat venography to assess the long term objective response. After 12 months, subjects having Implanon inserted were requested to rate their overall degree of satisfaction with therapy. RESULTS: All 25 women recruited in the study completed follow-up. Two cases were excluded from the study and referred to the psychiatry department after a negative evaluation for disease and HADS scores relevant for depression. An improvement in symptoms was observed throughout the 12 months amongst the Implanon group versus no treatment. The greatest changes in pain assessed using either the VAS or VRS were between the pretreatment scores and those after 6 months (7.7 +/- 1.3 vs. 4.6 +/- 3.0 for VAS, P < 0.001; and 25 +/- 13.8 vs. 19 +/- 18.9 for VRS, P < 0.002). The monthly quantified blood loss fell from 204 (196) pretreatment to 90 (157) at 6 months (P < 0.001) and then to 64 (32) at 9 months (P < 0.002). Objective repeat venography score was reduced significantly at 1 year after treatment compared with the baseline evaluation as well as with the control group (4.5 +/- 1.2 vs. 8.6 +/- 0.5; P = 0.001 and 4.2 +/- 0.9 vs. 8.5 +/- 0.6; P = 0.0002, respectively). At final satisfaction assessment, 2 (17%) women were very satisfied 8 (66%) were satisfied, and 2 (17%) were uncertain. The implant was retained by all women at the end of the study. CONCLUSION: Implanon seems to be an effective hormonal alternative for long-term treatment of properly selected patients with pure PCS-related pelvic pain. PMID- 19190928 TI - A randomized controlled trial comparing powdery sublingual misoprostol and sublingual misoprostol tablet for management of embryonic death or anembryonic pregnancy. AB - OBJECTIVES: To compare complete abortion rate, duration of abortion, and side effects between 600 microg powdery sublingual misoprostol and 600 microg sublingual misoprostol tablet for management of embryonic death or anembryonic pregnancy. MATERIALS AND METHODS: Fifty-four pregnant women up to 13 weeks of gestation diagnosed with embryonic death or anembryonic pregnancy were randomized to receive 600 microg powdery sublingual misoprostol or 600 microg sublingual misoprostol tablet. Complete abortion was evaluated by transvaginal ultrasound at 48 h. RESULTS: Twenty-six patients received 600 microg powdery sublingual misoprostol and 28 patients received 600 microg sublingual misoprostol tablet. Complete abortion rate was 34.6% in powdery sublingual misoprostol group and 32.1% in sublingual misoprostol tablet group (P = 0.847). Duration of abortion in powdery sublingual misoprostol group and sublingual misoprostol tablet group was similar (34.7 +/- 18.8 vs. 36.9 +/- 17.8 h, respectively, P = 0.656). There was no significant difference in the side effects between both groups. CONCLUSIONS: Single dose of 600 microg of powdery sublingual misoprostol does not improve its efficacy for management of embryonic death or anembryonic pregnancy when compared to sublingual misoprostol tablet. PMID- 19190930 TI - Alterations of epithelial adhesion molecules and basement membrane components in lattice corneal dystrophy (LCD). AB - BACKGROUND: The aim of the study was to investigate the histopathological and ultrastructural correlate of delayed epithelial healing in eyes with lattice corneal dystrophy (LCD). MATERIALS AND METHODS: Corneal buttons from 4 patients with LCD (two with subepithelial, two with stromal amyloid deposits) and 2 control corneas were examined. Cell-matrix adhesion molecules and basement membrane components of the corneal epithelium were analyzed by immunohistochemistry and hemidesmosomes between epithelium and stroma were quantified by transmission electron microscopy (TEM). RESULTS: By TEM well developed hemidesmosomes anchored the basal epithelial cells to the underlying basement membrane in all normal and LCD corneas. Hemidesmosome density was not significantly different in subepithelial (224.7 +/- 34.1/100 microm) and stromal (234.3 +/- 36.3/100 microm) LCD compared to controls (241.3 +/- 26.8/100 microm). The basement membrane was interrupted in subepithelial, but continuous in stromal LCD. Integrin alpha6 and beta4 staining formed a continuous line along the basal surface of the corneal epithelium in control corneas, whereas it appeared discontinuous and patchy both in subepithelial and stromal forms of LCD. Staining for alphaV integrin showed irregular staining patterns, i.e. enhanced labelling intensity in subepithelial and interrupted pattern in stromal LCD, respectively. Integrins alpha3, beta1, beta2, and beta5, dystroglycan, and plectin were not markedly different in dystrophic corneas. Type VII collagen showed a discontinuous staining in subepithelial forms of LCD. In stromal forms of LCD, type VII collagen staining occurred in additional patches underneath the epithelial basement membrane zone. Type XVII collagen staining was reduced in subepithelial LCD. Laminin-1, laminin-5 and laminin gamma2 showed variable irregular staining patterns in dystrophic corneas with focal interruptions, focal thickenings, and reduplications of basement membrane. Some irregularities in corneas with subepithelial amyloid were observed for collagen types IV, V, and XVIII, laminin alpha1, alpha3, and gamma1, nidogen-1 and -2, perlecan, fibrillin 1. CONCLUSIONS: Immunohistochemical and electron microscopic evidence of structural alterations was found in LCD compared to normal corneas concerning cell-matrix adhesion molecules and basement membrane components. These alterations were more pronounced in dystrophic corneas with subepithelial amyloid deposits than in those with stromal deposits. Histopathological findings may correspond to reduced cell-matrix interactions and partly explain delayed epithelial healing in patients with lattice corneal dystrophy. PMID- 19190931 TI - Cancer incidence in the wastewater treatment plant of a large chemical company. AB - OBJECTIVES: To evaluate cancer incidence among employees assigned to BASF's wastewater treatment plant. METHODS: We conducted a retrospective cohort study including 477 male employees who had ever worked in the facility for at least 1 year since the start of operations in 1974. Cancers were identified by review of occupational medical records and a standardized questionnaire completed by the participants or their next of kin. Confirmation through hospital records was sought for all reported cases after obtaining informed consent. Standardized incidence ratios (SIR) and 95% confidence intervals (CI) were computed using comparison data provided by the Saarland Cancer Registry. Further comparisons were made between three different subgroups of employees, working in maintenance, wastewater processing, and sewage sludge treatment. RESULTS: A total of 50 cancers were observed (SIR 1.14, CI 0.85-1.51). Colorectal (1.14, 0.42-2.48), bronchial (1.40, 0.67-2.57) and prostate (1.15, 0.42-2.50) were the most frequently observed cancers. Five cases of bladder cancer were found in the total cohort (1.75, 0.57-4.09), with four of them occurring in the sewage sludge treatment area (6.82, 1.86-17.46). Allowing for a 10-year lag did not significantly change the results. CONCLUSIONS: The overall cancer experience among employees of the wastewater treatment plant was similar to that of the corresponding general population. The finding of an excess risk for bladder cancer in one subgroup of workers was unexpected with regard to the available literature. There is no straightforward explanation for this finding, and it may be due to chance. An extended follow-up of this cohort will take place after 5 years. Annual bladder cancer screening is offered to active and retired employees from this plant for the time being. The current working conditions and work practices have been re-assessed by occupational hygienists and deemed to be safe. PMID- 19190932 TI - Salivary alpha-amylase, cortisol and chromogranin A responses to a lecture: impact of sex. AB - The aim of this study was to (1) examine the presence of stress on professors when they teach in front of 200 students and analyse objectively such stress using biomarkers such as salivary cortisol, chromogranin A (CgA) and alpha amylase (AA) (2) investigate whether sex affects the reactivity of salivary alpha amylase (sAA) and cortisol concentrations and the interaction of both hormonal systems. Fifty-two participants (26 women and 26 men) collected nine unstimulated saliva samples on 2 days, (one working day, and one resting day). Cortisol concentrations and AA activity measured on the teaching day were significantly higher than those noted on the resting values. No differences between the resting day and the teaching day were reported for CgA. Our results showed a cortisol response to teaching, which was characterized by an anticipatory rise. The AA level was significantly increased after the end of the lecture, and returned to the pre-lecture level 30 min after the end of the lecture. The awakening cortisol response noted on the teaching day was significantly higher in females than in males. No baseline sex differences in sAA and CgA were observed and men and women seem to have a comparable reactivity in sAA, CgA and cortisol levels on lecture stress. The mechanisms that leads to modify activity of salivary AA and CgA due to stress is not entirely understood and further research is needed to elucidate them. PMID- 19190933 TI - Tissue microarrays for immunohistochemical determination of oncological biomarkers. PMID- 19190934 TI - Spatial and temporal differences in the blood parasite fauna of passerine birds during the spring migration in Bulgaria. AB - Species composition and prevalence of the blood parasites of three migratory Acrocephalus species of warblers (Acrocephalus arundinaceus, Acrocephalus scirpaceus and Acrocephalus schoenobaenus) were studied during spring period in Bulgaria. For each of the studied species, we compared the parasite fauna during their spring migration at three study sites in search of infection patterns in time and space. Spatial differences were registered in the blood parasites and their prevalence for two of the studied species-A. arundinaceus and A. scirpaceus, whereas for the third one-A. schoenobaenus, there were no significant differences. The parasite species Haemoproteus belopolskyi was present in migrating A. arundinaceus only at Kalimok station, but not at the other two sampling locations. Plasmodium prevalence in A. arundinaceus migrating through Kalimok was also different between years. A decrease of H. belopolskyi prevalence was detected for A. schoenobaenus at Petarch during the spring of 2006. On the contrary, Plasmodium (Haemamoeba) sp. infections of the same bird species increased during the spring period. The differences found between sites could be explained with the different origin of the migrants at different migration stopover points. Climatic conditions and vector abundance in the zones of origin are probable reasons for the observed differences between years. PMID- 19190936 TI - What do we know about chronic renal failure in young adults? I. Primary renal disease. AB - Paediatric registries worldwide report that congenital abnormalities of the kidney and urinary tract (CAKUT) account for approximately 50% of end-stage renal failure and other congenital and familial diseases account for another 20% (together 70%). Does the same hold true for young adults? Almost nothing has been published about primary renal disease in adults who have reached end-stage before 30 years of age. I have reviewed the UK renal registry (2000-2006) and the United States Renal Data System (USRDS) data base (2005) to answer this question. While paediatric registries have reduced the number of children with 'no specific diagnosis' from 39% in 1976 to fewer than 5%, the adult registries still report rates of 20-27%, which rise to 28-36% when all unspecified groups, predominantly 'glomerulonephritis (GN) (histologically not examined)', are considered together. For UK data, this rise in 'no specific diagnosis' mirrors a fall in CAKUT to 26% for the age group 18-21 years. According to USRDS data, CAKUT falls from 31% for ages 0-19 years to only 5% for ages 20-30 years. Nephrologists probably under diagnose CAKUT in young adult patients, and this diagnosis can account for many of the 30% that currently have no specified primary renal disease. PMID- 19190935 TI - Dialysis-associated peritonitis in children. AB - Peritonitis remains a frequent complication of peritoneal dialysis in children and is the most common reason for technique failure. The microbiology is characterized by a predominance of Gram-positive organisms, with fungi responsible for less than 5% of episodes. Data collected by the International Pediatric Peritonitis Registry have revealed a worldwide variation in the bacterial etiology of peritonitis, as well as in the rate of culture-negative peritonitis. Risk factors for infection include young age, the absence of prophylactic antibiotics at catheter placement, spiking of dialysis bags, and the presence of a catheter exit-site or tunnel infection. Clinical symptoms at presentation are somewhat organism specific and can be objectively assessed with a Disease Severity Score. Whereas recommendations for empiric antibiotic therapy in children have been published by the International Society of Peritoneal Dialysis, epidemiologic data and antibiotic susceptibility data suggest that it may be desirable to take the patient- and center-specific history of microorganisms and their sensitivity patterns into account when prescribing initial therapy. The vast majority of patients are treated successfully and continue peritoneal dialysis, with the poorest outcome noted in patients with peritonitis secondary to Gram-negative organisms or fungi and in those with a relapsing infection. PMID- 19190937 TI - What do we know about chronic renal failure in young adults? II. Adult outcome of pediatric renal disease. AB - Congenital abnormalities of the kidney and urinary tract (CAKUT) account for more than half of all renal failure in children. For young adults with CAKUT two questions are paramount: what is the prognosis and what is the best management to improve outcome? The paediatric literature shows that prognostic factors are glomerular filtration rate (GFR) and the presence of proteinuria. We reviewed data from 101 young adult patients with either primary vesico-ureteric reflux and renal dysplasia or obstructive uropathy. Patients had an estimated GFR (eGFR) of 35 ml/min. The ACEI benefit increased with time. Rate of decline was slower than reported for other diseases and was only -2.4 ml/min per year for those reaching the start of dialysis. Outcome is predictable by the level of residual renal function (GFR). Nevertheless, function remains stable while proteinuria is minimal. Short-term studies overestimate rates of deterioration. PMID- 19190939 TI - The crystal structure of auracyanin A at 1.85 A resolution: the structures and functions of auracyanins A and B, two almost identical "blue" copper proteins, in the photosynthetic bacterium Chloroflexus aurantiacus. AB - Auracyanins A and B are two closely similar "blue" copper proteins produced by the filamentous anoxygenic phototrophic bacterium Chloroflexus aurantiacus. Both proteins have a water-soluble 140-residue globular domain, which is preceded in the sequence by an N-terminal tail. The globular domains of auracyanins A and B have sequences that are 38% identical. The sequences of the N-terminal tails, on the other hand, are distinctly different, suggesting that auracyanins A and B occupy different membrane sites and have different functions. The crystal structure of auracyanin A has been solved and refined at 1.85 A resolution. The polypeptide fold is similar to that of auracyanin B (Bond et al. in J Mol Biol 306:47-67, 2001), but the distribution of charged and polar residues on the molecular surface is different. The Cu-site dimensions of the two auracyanins are identical. This is unexpected, since auracyanin A has a shorter polypeptide loop between two of the Cu-binding residues, and the two proteins have significantly different EPR, UV-visible and resonance Raman spectra. The genes for the globular domains of auracyanins A and B have been cloned in a bacterial expression system, enabling purification of large quantities of protein. It is shown that auracyanin A is expressed only when C. aurantiacus cells are grown in light, whereas auracyanin B is expressed under dark as well as light conditions. The inference is that auracyanin A has a function in photosynthesis, and that auracyanin B has a function in aerobic respiration. PMID- 19190940 TI - Metastatic involvement of parotid from carcinoma of the breast--a case report. AB - INTRODUCTION: Involvement of parotid gland by metastases from distant primary malignant tumors is rare. CASE REPORT: A 42-year-old female having right breast cancer had reported to the Department of Oral Diagnosis and Medicine with a chief complaint of a swelling in the left parotid gland region for 2 months. On the basis of clinical findings, the primary malignancy of parotid gland was provisionally suspected. DISCUSSION: Microscopic examination of parotid and breast revealed that parotid gland was involved as a distant metastases from a breast carcinoma, indicating that a swelling in the parotid needs to be assessed and investigated very carefully. PMID- 19190941 TI - Establishing a surveillance network for severe lower respiratory tract infections in Korean infants and young children. AB - To reduce morbidity and mortality through integrated case management, a pilot study to detect respiratory viruses in patients with acute lower respiratory infections (ALRIs) was designed as part of a nationwide surveillance for this disease in Korea. The study population consisted of hospitalized patients under the age of 5 years with bronchiolitis, pneumonia, croup, or acute respiratory distress syndrome. A prospective 6-month study was performed. Two hundred and ninety-seven nasopharyngeal secretions were collected and multiplex reverse transcriptase polymerase chain reactions (RT-PCR)/polymerase chain reactions (PCR) were performed to detect respiratory viruses. If there were any positive RT PCR/PCR results, viral cultures were proceeded for confirmation. Respiratory viruses were identified in 49.6% of 296 patients. The detection rates were as follows: respiratory syncytial virus (RSV) was the most commonly detected in 52.7% (87/165), human metapneumovirus (hMPV) in 15.8%, human corona virus (hCoV) in 5.5%, adenovirus in 9.7%, human bocavirus (hBoV) in 5.5%, parainfluenza virus (PIV) in 3.6%, rhinovirus (RV) in 4.2%, and the influenza virus in 3% of the patients with ALRIs. The consistent rate of positive results between RT-PCR and viral culture was 92% (105/114). Using our methods to detect viral causes seemed to be acceptable for the national surveillance of severe acute respiratory infections in infants and children. PMID- 19190942 TI - Device-independent, real-time identification of bacterial pathogens with a metal oxide-based olfactory sensor. AB - A novel olfactory method for bacterial species identification using an electronic nose device called the MonoNose was developed. Differential speciation of micro organisms present in primary cultures of clinical samples could be performed by real-time identification of volatile organic compounds (VOCs) produced during microbial replication. Kinetic measurements show that the dynamic changes in headspace gas composition are orders of magnitude larger than the static differences at the end of fermentation. Eleven different, clinically relevant bacterial species were included in this study. For each of the species, two to eight different strains were used to take intra-species biodiversity into account. A total of 52 different strains were measured in an incubator at 37 degrees C. The results show that the diagnostic specificities varied from 100% for Clostridium difficile to 67% for Enterobacter cloacae with an overall average of 87%. Pathogen identification with a MonoNose can be achieved within 6-8 h of inoculation of the culture broths. The diagnostic specificity can be improved by broth modification to improve the VOC production of the pathogens involved. PMID- 19190943 TI - Evidence of intra-familial transmission of Helicobacter pylori by PCR-based RAPD fingerprinting in Bangladesh. AB - Helicobacter pylori is a genetically diverse bacterial species, which has facilitated adaptation to new hosts and persists worldwide. The main objective of this study was to explore intra-familial transmission of H. pylori in Bangladesh. We characterized H. pylori in 35 families including 138 family members using random amplified polymorphic DNA (RAPD) fingerprinting. Forty-six percent of H. pylori isolated from the mother shared a related genotype with strains isolated from their children. Twenty-nine percent of H. pylori isolates of the mother are related to the youngest children. Only 6% of the parents shared related genotype of H. pylori. These findings suggest that mother-to-child transmission occurs in early childhood and is the most probable route of transmission of H. pylori in Bangladesh. PMID- 19190945 TI - Discotic molecules in cylindrical nanopores: a Monte Carlo study. AB - We report Monte Carlo simulations of a model discotic molecule embedded in cylindrical pores. We consider a planar anchoring of the molecules on the surface for two different cylinder radii: R(*) = 5 and R(*) = 10 , in units of the molecular diameter. For both radii, we note that the system is progressively structured in concentric shells when decreasing the temperature. With the small radius, we observe continuous transitions from an isotropic to a nematic phase and then to a crystal one. The radius of the pores is sufficiently small to force the crystal to grow along their main axis. However some orientational discrepancies are observed: some samples present a zigzag configuration. With the big radius, the situation is more complex and it is likely that different scenarios are available. The crystals can be built along the main axis of the cylinders, as for the small radius, but also in any other direction. Thus we observe samples with different orientational domains. In the case of crystals oriented along the nanopore axis, we note that only the first 5 shells close to the wall are sensitive to it. PMID- 19190944 TI - Membrane transporters and carbon metabolism implicated in chloride homeostasis differentiate salt stress responses in tolerant and sensitive Citrus rootstocks. AB - Salinity tolerance in Citrus is strongly related to leaf chloride accumulation. Both chloride homeostasis and specific genetic responses to Cl(-) toxicity are issues scarcely investigated in plants. To discriminate the transcriptomic network related to Cl(-) toxicity and salinity tolerance, we have used two Cl(-) salt treatments (NaCl and KCl) to perform a comparative microarray approach on two Citrus genotypes, the salt-sensitive Carrizo citrange, a poor Cl(-) excluder, and the tolerant Cleopatra mandarin, an efficient Cl(-) excluder. The data indicated that Cl(-) toxicity, rather than Na(+) toxicity and/or the concomitant osmotic perturbation, is the primary factor involved in the molecular responses of citrus plant leaves to salinity. A number of uncharacterized membrane transporter genes, like NRT1-2, were differentially regulated in the tolerant and the sensitive genotypes, suggesting its potential implication in Cl(-) homeostasis. Analyses of enriched functional categories showed that the tolerant rootstock induced wider stress responses in gene expression while repressing central metabolic processes such as photosynthesis and carbon utilization. These features were in agreement with phenotypic changes in the patterns of photosynthesis, transpiration, and stomatal conductance and support the concept that regulation of transpiration and its associated metabolic adjustments configure an adaptive response to salinity that reduces Cl(-) accumulation in the tolerant genotype. PMID- 19190946 TI - Permeability of aqueous foams. AB - We perform forced-drainage experiments in aqueous foams and compare the results with data available in the literature. We show that all the data can be accurately compared together if the dimensionless permeability of the foam is plotted as a function of liquid fraction. Using this set of coordinates highlights the fact that a large part of the published experimental results corresponds to relatively wet foams (epsilon approximately 0.1). Yet, most of the foam drainage models are based on geometrical considerations only valid for dry foams. We therefore discuss the range of validity of the different models in the literature and their comparison to experimental data. We propose extensions of these models considering the geometry of foam in the relatively wet-foam limit. We eventually show that if the foam geometry is correctly described, forced drainage experiments can be understood using a unique parameter --the Boussinesq number. PMID- 19190947 TI - Structural, electrical, electronic and optical properties of melanin films. AB - We present thick, uniform and rather flat melanin films obtained using spray deposition. The morphology of the films was investigated using Scanning Electron Microscopy (SEM) and Atomic Force Microscopy (AFM). Temperature-dependent electrical resistance of melanin thin films evidenced a semiconductor-like character and a hysteretic behavior linked to an irreversible process of water molecule desorption from the melanin film. X-ray Photoelectron Spectroscopy (XPS) was carried out to analyze the role of the functional groups in the primary and secondary structure of the macromolecule, showing that the contribution of the 5,6-dihydroxyindole-2-carboxylic acid (DHICA) subunit to the molecule is about 35%. Comparison of the optical absorption of the thick (800nm) and thin (80nm) films showed a spectral change when the thickness increases. From in vacuum photoconductivity (PC) measured at controlled temperatures, we suggest that the melanin films exhibit a possible charge transport mechanism by means of delocalized pi states along the stacked planar secondary structure. PMID- 19190948 TI - The production of hydrogen sulphide and other aroma compounds by wine strains of Saccharomyces cerevisiae in synthetic media with different nitrogen concentrations. AB - The aim of this study was to evaluate the combined effect of initial nitrogen content on the production of hydrogen sulphide and other volatile compounds during alcoholic fermentation. For that propose, three commercial wine strains of Saccharomyces cerevisiae were used to ferment synthetic grape juice media under different nitrogen concentrations. H(2)S was measured throughout fermentations and other aroma compounds were analyzed at the end of the experiments. The trigger levels at which an inverse relationship between the initial nitrogen present in media and total H(2)S production varied among the three strains tested. For UCD522 and PYCC4072, the highest H(2)S levels were produced in media with 267 mg N l(-1) of initial nitrogen, whereas the lowest levels were detected with nitrogen limitation/starvation conditions (66 mg N l(-1)). Moreover, 21 other aroma compounds belonging to different chemical classes were identified and quantified by solid phase micro-extraction (SPME) coupled to gas chromatography mass spectrometry (GC-MS). The initial nitrogen concentration more than yeast strain had a decisive effect on the final aroma composition, suggesting that modulation of nutrients emerges as a useful tool for producing desired flavour and odour compounds. PMID- 19190949 TI - Indigenous and inoculated yeast fermentation of gabiroba (Campomanesia pubescens) pulp for fruit wine production. AB - The objectives of this study were to evaluate the potential of gabiroba Campomanesia pubescens (DC) O. Berg in the production of a beverage fermented using selected and wild yeasts from indigenous fermentation, analyze the volatile compounds profile present during the process of fermentation, and evaluate the sensory quality of the final beverage produced. Throughout the process of fermentation, when Saccharomyces cerevisiae UFLA CA 1162 was inoculated, there were stable viable populations around 9 log cells ml(-1). During indigenous fermentation, yeast population increased from 3.7 log CFU ml(-1) to 8.1 log CFU ml(-1) after 14 days. The diversity and dynamics of the yeast population during indigenous fermentation observed by PFGE analysis showed five different karyotyping profiles in the first days of fermentation. After the seventh day, there was a higher frequency of a similar S. cerevisiae profile. The yeast non Saccharomyces were identified by sequencing of the ITS region as Candida quercitrusa and Issatchenkia terricola. Inoculated fermentations yielded a higher amount of alcohol than indigenous ones, indicating the efficiency of selected strains. There was also a greater concentration of higher alcohols, which are usually responsible for the flavor found in alcoholic beverages. Based on the characteristics of the pulp and acceptance in the sensory analysis, gabiroba fruits showed good potential for use in the production of fermented beverage. PMID- 19190951 TI - Regulation of platelet glycoprotein VI (GPVI) surface expression and of soluble GPVI in patients with atrial fibrillation (AF) and acute coronary syndrome (ACS). AB - BACKGROUND: The platelet collagen receptor glycoprotein VI (GPVI) mediates platelet adhesion to subendothelial matrix and thrombus formation in acute coronary syndrome (ACS). This study examined patients with both ACS and stable coronary artery disease (CAD), which presented with atrial fibrillation (AF) and sinus rhythm (SR). METHODS AND RESULTS: We evaluated 992 patients with acute or stable CAD, and determined platelet surface expression of GPVI using flow cytometry. Seventy-eight patients presented with nonvalvular persistent AF. After 1:1 propensity score matching 156 matched cases with 78 pairs were obtained. Patients with AF and ACS showed a significantly decreased GPVI expression compared to patients with ACS and SR, whereas patients with stable angina pectoris (SA) presented with low level activation and no significant difference between SR and AF [mean fluorescence intensity (MFI) for ACS (SR Vs. AF): 20 +/- 6.3 Vs. 17.7 +/- 4.4; P = 0.023; SA (SR Vs. AF): 18.8 +/- 9.4 Vs. 18.1 +/- 6.1; P = 0.649]. In contrast, soluble GPVI was increased in ACS and AF accordingly [plasma GPVI (ng/ml) for ACS (SR Vs. AF): 1.4 +/- 0.8 Vs. 1.9 +/- 1.1; P = 0.038; SA (SR Vs. AF): 0.9 +/- 0.4 Vs. 1.1 +/- 0.5; P = 0.127]. CONCLUSION: Platelet GPVI surface expression is decreased in patients with AF and ACS compared to patients with SR and ACS. Nonvalvular AF is related to indices of chronic platelet activation and might be responsible for a down-regulation of GPVI receptor density on platelets, while soluble GPVI was increased in ACS and AF accordingly. PMID- 19190950 TI - Protective effects of EPA and deleterious effects of DHA on eNOS activity in Ea hy 926 cultured with lysophosphatidylcholine. AB - Oxidized low density lipoprotein (Ox-LDL) is a well-established risk factor in atherosclerosis and lysophosphatidylcholine (LysoPtdCho) is considered to be one of the major atherogenic component of Ox-LDL. The purpose of this work was to investigate the effects of two membrane n-3 long chain polyunsaturated fatty acids (n-3 PUFAs), EPA (eicosapentaenoic acid) and DHA (docosahexaenoic acid) compared to n-6 PUFA, ARA (arachidonic acid), on the activation of endothelial NO synthase (eNOS) by histamine in Ea hy 926 endothelial cells incubated during 24 h in the presence or the absence of LysoPtdCho. DHA (50 muM) produced a ROS induction in cells and aggravated the LysoPtdCho-induced oxidative stress. It did not modify the basal eNOS activity but impaired the stimulation of eNOS induced by histamine and was unable to correct the deleterious effect of LysoPtdCho on histamine-stimulated eNOS activity or phosphorylation of Ser 1177. In contrast, EPA (90 muM) did not modify the ROS level produced in the presence or absence of LysoPtdCho or basal eNOS activity and the stimulating effect of histamine on eNOS. However, it diminished the deleterious effect of LysoPtdCho as well as on the histamine-stimulated eNOS activity on the phosphorylation on Ser 1177 of eNOS. The beneficial effect of EPA but not DHA on endothelial eNOS activity in Ea hy 926 could be also partially due to a slight decrease in membrane DHA content in EPA-treated cells. Consequently, the equilibrium between NO generated by eNOS and ROS due to oxidative stress could explain, in part, the beneficial effect of EPA on the development of cardiovascular diseases. By contrast ARA an n-6 PUFA was devoid of any effect on ROS generation or eNOS activity in the basal state or after histamine-induced stimulation. In vivo experiments should be undertaken to confirm these results. PMID- 19190952 TI - Effects of the NO donor sodium nitroprusside on oxygen consumption and energetics in rabbit myocardium. AB - Nitric oxide (NO) has influence on various cellular functions. Little is known of the influence of NO on myocardial energetics. In the present study oxygen consumption and mechanical parameters of isometrically contracting rabbit papillary muscles (1 Hz stimulation frequency) were investigated at varying interventions while maintaining physiological conditions (37 degrees C; 2.5 mM Ca(2+)) to study the effects of NO on energetics. The NO donor sodium nitroprusside (SNP) showed a negative inotropic effect. SNP decreased the maximal force in normal rabbit muscle strips by 30%, the force time integral (FTI) by 40% and the relaxation time by 20%. In addition the oxygen consumption decreased by 60%, a notably disproportional decrease compared to the mechanical parameters. Consequently, the economy as a ratio of FTI and oxygen consumption is significantly increased by SNP. In contrast the negative inotropic effect due to a reduction in extracellular Calcium (Ca(2+)) from 2.5 to 1.25 mM reduced FTI and oxygen consumption proportionally by 40% and did not change economy. The effect of NO on force and oxygen consumption could be reproduced by the application of the cyclic guanosine monophosphate (cGMP) analogue 8-bromo-cGMP. In summary, NO increased the economy of isometrically contracting papillary muscles. The improvement in contraction economy under NO seems to be mediated by cGMP as the secondary messenger and maybe due to alterations of the crossbridge cycle. PMID- 19190953 TI - Changes in cellular contractility and cytokines profile during Trypanosoma cruzi infection in mice. AB - Trypanosoma cruzi, an intracellular protozoan parasite infecting a wide variety of vertebrates, is the agent responsible for Chagas' disease. This pathology often results in severe inflammatory heart condition and it is one of the major causes of dilated cardiomyopathy leading to heart failure in Latin America. Nevertheless, little is known about the changes in isolate cardiac myocytes contractility during the development of this pathology. Here we report a relationship between cytokines profile of mice infected with T. cruzi and the modifications in the cellular contractility pattern. We found that cellular contractility, measured as fractional shortening, showed a complex behavior. The changes were evaluated during the acute phase (15, 30 and 45 dpi) and chronic phase (>90 dpi). The time to half contraction and relaxation were lengthier despite the number of days after infection or the heart region evaluated. The maximal contraction and relaxation velocities were significantly slower. The observed changes in cellular contractility were correlated with the presence of circulating IFN-gamma, TNF-alpha and MCP-1/CCL2 during the course of infection. Together, our data demonstrate that cellular contractility is altered in the three heart regions studied, and these alterations are observed at the very beginning of the parasitism and they remained until the chronic phase has been reached. Indeed, we propose a role for IFN-gamma, TNF-alpha and MCP-1/CCL2 in the mechanical heart remodeling during experimental Chagas' disease. PMID- 19190955 TI - Ischemic injury activates PTHrP and PTH1R expression in human ventricular cardiomyocytes. AB - The PTHrP/PTH1R signalling system induces calciotropic and myorelaxant effects on the vascular system and plays critical roles in the development of the cardiovascular system. In animal models, PTHrP exerts important effects on heart rate and contractility, particularly under ischemic conditions, while, in human hearts, the expression of PTHrP by cardiomyocytes remains to be defined in both normal and ischemic conditions. The present study has been conducted on 101 autoptical cases and confirmed on nine explanted hearts in order to analyze the expression of the PTHrP/PTH1R system by ventricular myocardium in respect to morphological aspects of the myocardial ischemic damage, myofiber hypertrophy and disarray, coronarosclerosis, age and sex. Immunohistochemistry showed positive cytoplasmic immunostaining for both PTHrP and PTH1R in ventricular cardiomyocytes. The expression levels of the PTHrP/PTH1R system resulted significantly increased (P = 0.0008 and P < 0.0001, respectively) in association with the myocardial ischemic damage and the presence of cardiomyocyte hypertrophy (P = 0.02 and P = 0.009 respectively). Conversely, increased expression levels of PTHrP alone were observed in myofiber disarray (P = 0.04), whereas PTH1R was augmented in coronarosclerosis (P = 0.004) and age (P = 0.001). Taken together, these results demonstrate that human ventricular cardiomyocytes express PTHrP and PTH1R and suggest that the activation of the PTHrP/PTH1R system could represent an aspect of the embryonic gene program typically reactivated by the myocardium when subjected to ischemia and/or hypertrophy. PMID- 19190956 TI - Impact of myocardial inflammation on cytosolic and mitochondrial creatine kinase activity and expression. AB - The disturbance of myocardial energy metabolism has been discussed as contributing to the progression of heart failure. Little however is known about the cardiac mitochondrial/cytosolic energy transfer in murine and human inflammatory heart disease. We examined the myocardial creatine kinase (CK) system, which connects mitochondrial ATP-producing and cytosolic ATP-consuming processes and is thus of central importance to the cellular energy homeostasis. The time course of expression and enzymatic activity of mitochondrial (mtCK) and cytosolic CK (cytCK) was investigated in Coxsackievirus B3 (CVB3)-infected SWR mice, which are susceptible to the development of chronic myocarditis. In addition, cytCK activity and isoform expression were analyzed in biopsies from patients with chronic inflammatory heart disease (n = 22). Cardiac CVB3 titer in CVB3-infected mice reached its maximum at 4 days post-infection (pi) and became undetectable at 28 days pi; cardiac inflammation cumulated 14 days pi but persisted through the 28-day survey. MtCK enzymatic activity was reduced by 40% without a concurrent decrease in mtCK protein during early and acute MC. Impaired mtCK activity was correlated with virus replication and increased level of interleukine 1beta (IL-1beta), tumor necrosis factor alpha (TNFalpha), and elevated catalase expression, a marker for intracellular oxidative stress. A reduction in cytCK activity of 48% was observed at day 14 pi and persisted to day 28 pi. This restriction was caused by a decrease in cytCK subunit expression but also by direct inhibition of specific cytCK activity. CytCK activity and expression were also reduced in myocardial biopsies from enterovirus genome negative patients with inflammatory heart disease. The decrease in cytCK activity correlated with the number of infiltrating macrophages. Thus, viral infection and myocardial inflammation significantly influence the myocardial CK system via restriction of specific CK activity and down-regulation of cytCK protein. These changes may contribute to the progression of chronic inflammatory heart disease and malfunction of the heart. PMID- 19190954 TI - Vascular control in humans: focus on the coronary microcirculation. AB - Myocardial perfusion is regulated by a variety of factors that influence arteriolar vasomotor tone. An understanding of the physiological and pathophysiological factors that modulate coronary blood flow provides the basis for the judicious use of medications for the treatment of patients with coronary artery disease. Vasomotor properties of the coronary circulation vary among species. This review highlights the results of recent studies that examine the mechanisms by which the human coronary microcirculation is regulated in normal and disease states, focusing on diabetes. Multiple pathways responsible for myogenic constriction and flow-mediated dilation in human coronary arterioles are addressed. The important role of endothelium-derived hyperpolarizing factors, their interactions in mediating dilation, as well as speculation regarding the clinical significance are emphasized. Unique properties of coronary arterioles in human vs. other species are discussed. PMID- 19190957 TI - Bipolar I disorder with mood-incongruent psychotic symptoms: a comparative longitudinal study. AB - OBJECTIVES: The purpose of this paper is to demonstrate similarities and differences between bipolar I patients with and without mood-incongruent symptoms (MIS) over a long period of time, independently of longitudinal syndromatic constellations. METHODS: The Halle bipolarity longitudinal study (HABILOS) prospectively investigates 182 patients meeting the DSM-IV criteria for bipolar I disorders over a long period of time (x;- = 16.84 years). One thousand five hundred thirty-nine (1,539) episodes have been evaluated with standardized instruments. Patients and episodes were divided into two groups (with and without MIS) and were compared on various levels. RESULTS: It was found: (1) The majority of the episodes of bipolar I patients during long-term course did not have MIS, but the majority of patients did. (2) Bipolar I patients with MIS differ from patients without MIS in the following features: (a) Bipolar I patients with MIS are more frequently males. (b) Bipolar I patients with MIS need treatment at a significantly younger age than those without MIS. (c) First manifestation of bipolar I disorder with MIS after the age of 50 is extremely seldom. (d) Bipolar I patients with MIS more frequently have relatives with schizophrenia. (e) Bipolar I patients with MIS more frequently become disabled and retire at a significantly younger age than patients without MIS and (f) Significantly fewer patients with MIS than those without MIS live in a stable partnership. CONCLUSIONS: It can be concluded that bipolar I disorders with MIS are more severe disorders than bipolar I disorders without MIS. This finding in combination with the above results, however, can give rise to the conclusion that bipolar I disorders with MIS are the epiphenomenon of the overlap, possibly genetic, of a "schizophrenic spectrum" and a "bipolar spectrum" and their antagonistic influence creating a "schizo-affective" area between them as a kind of psychotic continuum between prototypes. PMID- 19190958 TI - A randomized controlled clinical trial of citalopram versus fluoxetine in children and adolescents with obsessive-compulsive disorder (OCD). AB - OBJECTIVE: Several controlled trials have demonstrated the efficacy and safety of Fluoxetine in children and adolescents with Obsessive-Compulsive Disorder (OCD), but there is no controlled study on the effectiveness of Citalopram in this group. This report describes the use of Citalopram in comparison with Fluoxetine in childhood-onset OCD. METHOD: This study is a randomized, double blind, fixed does (20mg) trial of Fluoxetine versus Citalopram in 29 children and adolescents (17 boys and 12 girls) with OCD, aged 7-18 years (mean 13.8 and SD 3.05). The length of study was 6 weeks. Obsessive-Compulsive symptom severity was measured by Yale-Brown Obsessive-Compulsive Scale (CY-BOCS) and Clinician's Global Impression Scale (CGI). DICA (Diagnostic Interview of Children and Adolescents) was used to diagnose the psychiatric disorders. RESULTS: Each group showed significant improvement over the baseline as measured by the CY-BOCS (p < 0.01) but not by CGI (p = NS). The Comparison between two groups showed no significant differences in efficacy and safety of the drugs. Most common adverse effects were headache for Citalopram and tremor for Fluoxetine. CONCLUSION: The results suggest that Citalopram is as safe and effective as Fluoxetine for children and adolescents with OCD. Further studies are needed to replicate our findings. PMID- 19190959 TI - Pupil evaluation as a test for autonomic disorders. AB - Pupil tests provide a convenient and simple method for evaluation of autonomic function. Most patients with autonomic disorders show evidence of sympathetic or parasympathetic deficits in the pupil, and these can be detected using a combination of clinical signs, pupillometric tests (measuring the responses to light, or an accommodative effort, or a sudden noise) and pharmacological tests (using topically applied drugs both to confirm a deficit and to localize the lesion). Caution is needed in the interpretation of these tests, particularly if the deficits are mixed (i.e. sympathetic and parasympathetic) or bilateral. The pattern of autonomic disturbance in the pupils often correlates poorly with autonomic function elsewhere, but may have diagnostic value in discriminating between different underlying conditions. PMID- 19190960 TI - Courvoisier's gallbladder: law or sign? AB - BACKGROUND: Variously described as Courvoisier's law, sign, or even gallbladder, this eponymous "law" has been taught to medical students since the publication of Courvoisier's treatise in 1890. METHODS: We reviewed Courvoisier's original "law," the modern misconceptions surrounding it, and the contemporary evidence supporting and explaining his observations. RESULTS: Courvoisier never stated a "law" in the context of a jaundiced patient with a palpable gallbladder. He described 187 cases of common bile duct obstruction, observing that gallbladder dilatation seldom occurred with stone obstruction of the bile duct. The classic explanation for Courvoisier's finding is based on the underlying pathologic process. With the presence of gallstones come repeated episodes of infection and subsequent fibrosis of the gallbladder. In the event that a gallstone causes the obstruction, the gallbladder is shrunken owing to fibrosis and is unlikely to be distensible and, hence, palpable. With other causes of obstruction, the gallbladder distends as a result of the back-pressure from obstructed bile flow. However, recent experiments show that gallbladders are equally distensible in vitro, irrespective of the pathology, suggesting that chronicity of the obstruction is the key. Chronically elevated intraductal pressures are more likely to develop with malignant obstruction owing to the progressive nature of the disease. Gallstones cause obstruction in an intermittent fashion, which is generally not consistent enough to produce such a chronic rise in pressure. CONCLUSION: We hope that reminding clinicians of Courvoisier's actual observations will reestablish the usefulness of this clinical sign in the way he intended. PMID- 19190961 TI - Anatomic significance of a positive barium enema in deep infiltrating endometriosis of the large bowel. AB - PURPOSE: The anatomopathological significance of a positive double-contrast barium enema (DCBE) for suspicion of deep infiltrating endometriosis of the large bowel was studied. This is a retrospective study of a prospective database. METHODS: A large-bowel resection was proposed for patients who were suspicious for large-bowel endometriosis and had a positive DCBE. In a series of 73 patients, 71 large-bowel resections were performed. Histology and immunohistochemistry with the monoclonal antibody CD-10 were performed on the resection specimen. Outcome measures were the length of the resected specimen, the largest diameter of the lesion, the positivity of the resection margins, and the degree of infiltration of the large bowel. We also compared the mean largest diameters of the lesions with the degree of infiltration of the large bowel. RESULTS: Between December 1997 and October 2005, 80 patients were suspicious for large-bowel endometriosis: 73 (91%) had positive DCBEs, and 7 (9%) had negative DCBEs. Of the 73 with positive DCBEs, 4 (5%) refused digestive resection and 1 (1.4%) was excluded. Three patients underwent two large-bowel resections because of the presence of bifocal lesions (left and right colon). A total of 71 resections were performed. In case of positive DCBE, the perivisceral fat and the whole muscularis were infiltrated in 100% of cases. The infiltration reached the submucosa and the mucosa respectively in 82% and 18% of cases. A total of 9.9% of resection margins were positive at histology but only focally. The mean largest diameter of the lesions infiltrating the whole thickness of the large bowel was not statistically different from the mean largest diameter of more superficial lesions. CONCLUSIONS: Findings of mass effect with indentations and ridging of the mucosa on DCBE in a setting suspicious for large-bowel endometriosis correspond well with pathologic findings of deep infiltration of the large-bowel wall. Clinicians dealing with deep infiltrating endometriosis should be aware of these findings, which could influence their choice of surgical treatment. PMID- 19190962 TI - Differentiation of urinary stone and vascular calcifications on non-contrast CT images: an initial experience using computer aided diagnosis. AB - The purpose of this study was to develop methods for the differentiation of urinary stones and vascular calcifications using computer-aided diagnosis (CAD) of non-contrast computed tomography (CT) images. From May 2003 to February 2004, 56 patients that underwent a pre-contrast CT examination and subsequently diagnosed as ureter stones were included in the study. Fifty-nine ureter stones and 53 vascular calcifications on pre-contrast CT images of the patients were evaluated. The shapes of the lesions including disperseness, convex hull depth, and lobulation count were analyzed for patients with ureter stones and vascular calcifications. In addition, the internal textures including edge density, skewness, difference histogram variation (DHV), and the gray-level co-occurrence matrix moment were also evaluated for the patients. For evaluation of the diagnostic accuracy of the shape and texture features, an artificial neural network (ANN) and receiver operating characteristics curve (ROC) analyses were performed. Of the several shape factors, disperseness showed a statistical difference between ureter stones and vascular calcifications (p < 0.05). For the internal texture features, skewness and DHV showed statistical differences between ureter stones and vascular calcifications (p < 0.05). The performance of the ANN was evaluated by examining the area under the ROC curves (AUC, A (z)). The A (z) value was 0.85 for the shape parameters and 0.88 for the texture parameters. In this study, several parameters regarding shape and internal texture were statistically different between ureter stones and vascular calcifications. The use of CAD would make it possible to differentiate ureter stones from vascular calcifications by a comparison of these parameters. PMID- 19190963 TI - Effects of a Caenorhabditis elegans dauer pheromone ascaroside on physiology and signal transduction pathways. AB - Daumone is one of the three purified and artificially synthesized components of the Caenorhabditis elegans dauer pheromone. It affects the major signal transduction pathways known to discriminate between developmental arrest at the dauer stage and growth to the adult [the transforming growth factor beta (TGF beta) and daf-2/IGF1R pathways], just as natural pheromone extracts do. Transcription of daf-7/TGF-beta is reduced in pre-dauer larvae, and nuclear localization of the DAF-16/FOXO transcription factor is increased in embryos and L1 larvae exposed to synthetic daumone. However, daumone does not require the cilia in the amphidial neurons to produce these effects nor does it require the Galpha protein GPA-3 to induce dauer entry, although GPA-3 is required for dauer induction by natural dauer pheromone extracts. Synthetic daumone has physiological effects that have not been observed with natural pheromone. It is toxic at the concentrations required for bioassay and is lethal to mutants with defective cuticles. The molecular and physiological effects of daumone and natural dauer pheromone are only partially overlapping. PMID- 19190964 TI - Differentiating early and late recurrences after resection of HCC in cirrhotic patients: implications on surveillance, prevention, and treatment strategies. PMID- 19190965 TI - Circulating tumor cells in patients undergoing surgery for primary breast cancer: preliminary results of a pilot study. AB - INTRODUCTION: Circulating tumor cells (CTCs) have recently been shown to be an independent predictor of progression-free and overall survival in patients undergoing treatment for metastatic breast cancer. This study evaluates the presence and significance of CTCs in patient undergoing surgical resection of clinically localized primary breast cancer. METHODS: Patients undergoing surgery for clinically localized primary breast cancer were enrolled into a prospective study. Thirty milliliters of blood was drawn and studied using the CellSearch assay. RESULTS: Forty-one patients were enrolled at a single tertiary referral center. Ten patients (24.4%) had detectable CTCs preoperatively (PreOp). Nine (30%) patients were found to have CTCs postoperatively (PostOp). Overall, 16 (39%) were found to have CTCs either PreOp or PostOp. Hormone-negative patients were significantly more likely to have CTCs than hormone-positive patients. No other pathologic factor was predictive of the presence of CTCs. CONCLUSION: CTCs are detectable and quantifiable in breast surgery patients. CTCs were more likely to be found in hormone receptor negative patients. Further study will allow correlation with other pathological variables and clinical outcome. PMID- 19190966 TI - Lung cancer as a second primary malignancy: increasing prevalence and its influence on survival. AB - BACKGROUND: The purpose of this study was to review the experience with lung cancer as a second primary malignancy (LCSPM) and to determine whether a history of previous malignancy was an independent prognostic factor in resected non small cell lung cancer (NSCLC). METHODS: We included 414 patients undergoing pulmonary resection with a curative intention between January 1986 and January 2007. The criteria for classifying a lesion as a lung cancer as a second primary malignancy (LCSPM) were a different histological type or a tumor of identical histological type occurring more than 3 years after treatment of the primary tumor, or separated from the initial primary tumor by more than 2 cm of clinically normal epithelium. RESULTS: Thirty-four patients (8.2%) had a history of a previous malignancy. LCSPM were more frequent during the last decade (11.4% vs. 3.91%, P = .0009). Breast, uroepithelial, and lung cancer were the most prevalent initial primary tumors. In 64.7% of patients the time interval between the last malignancy and the current lung cancer was 2 years or shorter. Patients with NSCLC and a previous malignancy did not have a significantly different survival rate than patients with the current NSCLC as the first malignancy (5-year survival 65.3% vs. 58.6%, log-rank P = .416). CONCLUSION: The effectiveness of the surgical resection and the subsequent survival of these patients appear to be well demonstrated, and operative mortality and complications are no higher than for other lung cancer patients. PMID- 19190967 TI - Hepatic metastasectomy for testicular germ cell tumors: is it worth it? AB - BACKGROUND: Chemotherapy is highly effective for metastatic germ cell tumor (GCT), but experience with resection of hepatic metastases from GCT is limited. METHODS: Fifteen patients with GCT metastatic to the liver underwent 16 hepatic operations (1975-2002). Pre-resection therapy, surgical pathology, and operative outcomes were reviewed. All patients were followed to death or last contact for survival and disease status. RESULTS: Patients underwent biopsy (three), wedge resection (nine), bisegmentectomy (two), and major lobectomy (two). Hepatic histology included: necrosis (33%), viable tumor (27%), mature teratoma (13%), and benign histology (27%). Concomitant resection of extrahepatic disease (14 patients, 93%) found necrosis (53%), mature teratoma (27%), and viable tumor (13%). Operative mortality was 0% and morbidity was 40%. At 8.2 years (mean) from resection, 11 patients (73%) were alive: five with no evidence of disease, two with elevated tumor marker only, and four with gross disease. Four patients (27%) died. The 10-year overall survival was 62% from diagnosis. CONCLUSION: Resection of post-chemotherapy hepatic disease is safe, even when combined with resection of extrahepatic residual disease. The varied histologic findings, lack of reliable predictors, and prolonged survival achieved support a multidisciplinary approach which includes surgical resection of hepatic metastases. PMID- 19190968 TI - Implementation of a critical pathway for distal pancreatectomy at an academic institution. AB - OBJECTIVE: This study was designed to identify quantifiable parameters to track performance improvements brought about by the implementation of a critical pathway for complex alimentary tract surgery. BACKGROUND: Distal pancreatectomy is among the more complex general surgical procedures. This is primarily due to the possibility of blood loss from visceral vessels, splenic injury, and significant postoperative complications. The introduction of the laparoscopic approach to the distal pancreas has introduced a further level of surgical expertise required to fully address the clinical needs of this diverse patient population. Critical pathways have been one of the key tools used to achieve consistently excellent outcomes at high-quality, high-volume institutions. It remains to be determined if implementation of a critical pathway at an academic institution with prior moderate experience with distal pancreatectomy will result in performance gains and improved outcomes. METHODS: Between January 1, 2003 and August 15, 2007, 111 patients underwent distal pancreatectomy. Forty patients underwent resection during the 34-month period before the implementation of a critical pathway on October 15, 2005 and 71 during the 20 months after pathway implementation. Patients undergoing both open and laparoscopic procedures were included. Peri- and postoperative parameters were analyzed retrospectively to identify those that could be used to track performance improvement and outcomes. RESULTS: The two groups were not significantly different with respect to age, sex, race, diagnosis, operative blood loss, or mean operative duration. Postoperative length of hospital stay was significantly shorter when comparing pre- to postpathway implementation (10.2 days versus 6.7 days, P < or = 0.037). The rate of readmission to the hospital after discharge was significantly lower post pathway (25% versus 7%, P < or = 0.027). Hospital costs were also reduced. CONCLUSION: Implementation of a critical pathway for a complex procedure can be demonstrated to improve short-term outcomes at an academic institution. This improvement can be quantified and tracked and has implications for better utilization of resources and overall cost containment while maintaining or improving upon an already high level of care. PMID- 19190969 TI - Seasoned surgeons assessed in a laparoscopic surgical crisis. AB - OBJECTIVE: Maintenance of certification is a relatively new concept in the United States, and there is no mandatory retirement for surgeons. Our aim was to compare technical and team performance of surgeons of different ages in a simulated laparoscopic surgical crisis and validate a potential recredentialing tool for surgeons. METHODS: Using a single-blinded protocol, the performance of six "Seasoned" surgeons >55 years (mean 64, range 55-83) was compared to six "control" surgeons <55 years (mean 46, range 34-53) in a simulation. Surgical teams established pneumoperitoneum, trocar access, and managed intraabdominal hemorrhage in a simulated laparoscopic cholecystectomy while videotaped as part of an IRB protocol. Surgeons' performance was scored using validated technical and team performance scales. RESULTS: All of the "seasoned" surgeons relegated the use of unfamiliar technology to their assistants. All control surgeons achieved intraabdominal pneumoperitoneum themselves. Mean blood loss for seasoned surgeons and control surgeons was 2,555 versus 2,725 ml (NS), respectively. After recognition of bleeding in the unstable patient, senior surgeons converted to an urgent laparotomy case after 2.4 vs. 3.3 min for control group (NS). No difference was observed in overall technical and team abilities (p = NS). On debriefing, 85% of surgeons recommended simulation for training and recertification. CONCLUSIONS: Seasoned surgeons can use their assistant surgeon well to assure a safe and effective operation. Mandatory operating room retirement based on age may be arbitrary and should be replaced by performance measures. Simulation may prove a valuable tool for self -assessment and recredentialing. PMID- 19190970 TI - MicroRNA-143 and -205 expression in neosquamous esophageal epithelium following Argon plasma ablation of Barrett's esophagus. AB - INTRODUCTION: Ablation of Barrett's esophagus using Argon plasma coagulation (APC) is usually followed by the formation of a neosquamous epithelium. Investigating simple columnar or stratified squamous epithelium associated cytokeratin and microRNA (miRNA) expression in neo-squamous epithelium could help determine the identity and stability of the neosquamous epithelium. METHODS: Nine patients underwent ablation of Barrett's esophagus with APC. Biopsies were collected from Barrett's esophagus mucosa and proximal normal squamous epithelium before ablation, and from neosquamous and normal squamous epithelium after ablation. Additional esophageal mucosal biopsies from ten nonrefluxing subjects were used as a reference. RNA was extracted and real-time polymerase chain reaction was used to measure the expression of the cytokeratins CK-8 and CK-14 and the microRNAs miR-143 and miR-205. RESULTS: CK-8 and miR-143 expression were significantly higher in Barrett's esophagus mucosa, compared to neosquamous and normal squamous epithelium before and after APC, whereas miRNA-205 and CK-14 expression was significantly lower in Barrett's esophagus mucosa compared to all categories of squamous mucosa. The expression of CK-8, CK-14, miR-205, and miR 143 was similar between neosquamous epithelium compared to normal squamous epithelium in patients with Barrett's esophagus. Only miR-143 expression was significantly higher in neosquamous and normal squamous epithelium before and after APC compared to normal squamous epithelium from control subjects (p < 0.004). CONCLUSIONS: The expression levels of cytokeratins and miRNAs studied in post-ablation neosquamous epithelium and normal squamous epithelium in patients with Barrett's esophagus are similar. In patients with Barrett's esophagus, miR 143 expression is still elevated in both neosquamous mucosa, and the squamous mucosa above the metaplastic segment, suggesting that this mucosa may not be normal; i.e., it is different to that seen in subjects without Barrett's esophagus. miR-143 could promote a Barrett's epithelium gene expression pattern, and this could have a role in development of Barrett's esophagus. PMID- 19190971 TI - Is it possible to treat recurrent clubfoot with the Ponseti technique after posteromedial release?: a preliminary study. AB - The Ponseti technique for treating clubfoot has been popularized for idiopathic clubfoot and more recently several syndromic causes of clubfoot. We asked whether it could be used to treat recurrent clubfoot following failed posteromedial release. We retrospectively reviewed 58 children (83 clubfeet) treated by the Ponseti technique for recurrent deformity after posteromedial release in three centers. The minimum followup was 24 months (average, 45 months; range, 24-80 months). We determined initial and final Pirani scores and range of motion of the ankle and subtalar joint. Plantigrade and fully corrected feet were obtained in 71 feet (86%); 11 feet obtained partial correction; one patient failed treatment and underwent another posteromedial release. Recurrences occurred in nine patients (12 feet or 14%). Initial Pirani scores improved in all but one patient; severity of deformity was also inferred by number of casts used for treatment. The age at treatment and numbers of casts did not influence the scores of Pirani et al. The scores were similar among the three orthopaedic surgeons. LEVEL OF EVIDENCE: Level IV, therapeutic study. See Guidelines for Authors for a complete description of levels of evidence. PMID- 19190972 TI - Massive bone loss from fungal infection after anterior cruciate ligament arthroscopic reconstruction. AB - Although there are numerous reports of septic pyogenic arthritis after arthroscopic anterior cruciate ligament (ACL) reconstruction, there is limited information regarding the outcomes of fungal infection. We determined the outcomes of six patients with mycotic infection after regular ACL reconstruction. There were four males and two females with a mean age of 33 years. We determined the number of procedures performed, bone loss originating to control infection, and final reconstruction in these patients. An average of five arthroscopic lavage procedures had been performed at the referring centers. Fungal infection was diagnosed based on pathologic samples; five infections were the result of mucormycosis and one was Candida. After final debridement, the mean segmental bone loss was 12.8 cm. All patients were treated with intravenous antifungal coverage and cement spacers before final reconstruction. At final followup, all patients were free of clinical infection. Three had reconstruction with an allograft-prosthesis composite, two with hemicylindrical allografts, and one with an intercalary allograft arthrodesis. Despite the extremely unusual presentation of this complication, surgeons should be aware of potential and catastrophic consequences of this severe complication after ACL reconstruction. PMID- 19190973 TI - Novel measurement technique of the tibial slope on conventional MRI. AB - The posterior inclination of the tibial plateau, which is referred to as posterior tibial slope, is determined routinely on lateral radiographs. However, radiographically, it is not always possible to reliably recognize the lateral plateau, making a separate assessment of the medial and lateral plateaus difficult. We propose a technique to measure the plateaus separately by defining a tibial longitudinal axis on a conventional MRI. The medial plateau posterior tibial slope obtained from radiographs was compared with MR images in 100 consecutive patients with knee pain when ligament or meniscal injury was assumed. The posterior tibial slope on MRI correlated with those on radiographs. The mean posterior tibial slope was 3.4 degrees smaller on MRI compared with radiographs (4.8 degrees +/- 2.4 degrees versus 8.2 degrees +/- 2.8 degrees , respectively). The reproducibility was slightly better on radiographs than MRI (+/- 0.9 degrees versus +/- 1.4 degrees ). Twenty-one of the 100 cases had more than a 5 degrees difference (range, -8.7 degrees to 8.9 degrees ) between the medial and lateral plateaus. The proposed technique allows measurement of the posterior tibial slope of the medial and lateral plateaus on a standard knee MRI. By using this novel measurement technique, a reliable assessment of the medial and lateral tibial plateaus is possible. LEVEL OF EVIDENCE: Level III, diagnostic study. See the Guidelines for Authors for a complete description of levels of evidence. PMID- 19190974 TI - CT study on the effect of different treatment protocols for clubfoot pathology. AB - In congenital clubfoot, residual deformities are not well-documented and they may change depending on different treatments. To identify the treatment that provides better outcome at maturity, we studied the computed tomography of two cohorts of patients affected with congenital clubfoot who were treated using two distinct protocols. Forty-seven clubfeet were treated according to the traditional protocol of our hospital and 61 were treated according to the Ponseti technique. The normal feet of the unilateral deformities served as controls. All patients were followed to skeletal maturity. The ankle torsion angle and the declination angle of the neck of the talus were higher than normal but different only in patients treated with the traditional method. The calcaneocuboid angle was lower but only in patients treated with the Ponseti method. The shape of the talar joints was altered in many feet regardless of protocol. The CT images suggest the modifications of the torsion angle of the ankle, the declination angle of the neck of the talus, and the calcaneocuboid angle at maturity are related to the treatment protocol followed. The Ponseti manipulative technique provided better anatomical results in comparison to our traditional technique. PMID- 19190975 TI - The importance of population-specific normal database for quantification of myocardial ischemia: comparison between Japanese 360 and 180-degree databases and a US database. AB - OBJECTIVES: We aimed to develop a Japanese normal database for specific acquisition conditions, to compare US and Japanese normal populations, and to examine effects of camera rotation angle range on the normal limits. METHODS AND RESULTS: Stress-rest (99m)Tc myocardial perfusion databases for 360 degrees (Jp360) and 180 degrees (Jp180) acquisitions were created by the working group activity of the Japanese Society of Nuclear Medicine using Japanese patients. A standard 180 degrees database (US180) had been previously generated by the Cedars Sinai Medical Center based on American patients. Additionally, 90 Japanese patients underwent coronary arteriography and stress-rest (99m)Tc perfusion study with 360 degrees acquisition for validation purposes, and quantitative evaluation was performed by QPS software using the above three normal database sets. Major differences between US180 and Jp360 databases were found in the apex and in the anterior wall in females and in the inferior wall in males. When the diagnostic performance was evaluated by receiver-operating characteristic analysis, area under the curve was the highest for Jp360 (0.842), followed by Jp180 (0.758) and US180 (0.728) databases (P = .019, Jp360 vs US180; P = .035, Jp360 vs Jp180). The coronary territory score at stress was highest with the Jp360 database in male patients with right coronary artery stenosis (n = 26, Jp360: 4.92 +/- 4.61 [mean +/- SD], Jp180: 4.23 +/- 4.29, US180: 2.92 +/- 3.53; P < .0001 between Jp360 and US180) and in female patients with left anterior descending artery stenosis (n = 12, Jp360: 6.33 +/- 4.76, Jp180: 5.25 +/- 4.83, US180: 4.50 +/- 4.15; P = .0076 between Jp360 and US180). CONCLUSION: Because of the differences between US and Japanese normal databases, it is essential to use population- and acquisition specific databases when using quantitative perfusion SPECT software. PMID- 19190976 TI - Pharmacoepidemiologic research in Australia: challenges and opportunities for monitoring patients with rheumatic diseases. AB - The topic of drug safety has received great attention in recent years. Pharmacoepidemiology is the study of the use and effects of medicines in large populations using epidemiological methods. Pharmacoepidemiologic research can fill the knowledge gaps due to the limitations of existing pharmacovigilance systems that rely on randomised controlled trials and voluntary reporting. This review discusses the present state of pharmacoepidemiologic research in Australia. In Australia, linking administrative data on use of medications and medical services is possible to a certain extent. Data from patient registries with respect to rheumatology are also available. These data are valuable for better understanding of the beneficial and adverse effects of medicines. Opportunities and challenges of using these data sources to address issues from clinical pharmacology are also highlighted. Australia is well-placed internationally to make major contributions to the knowledge base of outcomes of medicines in the real-world setting. Developments in pharmacoepidemiology are critical to clinicians treating patients with rheumatic and other conditions. PMID- 19190978 TI - Resuscitation decisions in Irish long-stay units. AB - BACKGROUND: Little is known of how cardiopulmonary resuscitation (CPR) decisions are made in Irish long-term care settings. AIM: To examine how CPR decisions are made in Irish long-stay units and those factors associated with use or non-use of CPR. METHODS: We surveyed each public long-stay unit and a random sample of private nursing homes across the country. RESULTS: Of the 84 long-stay units that responded (response rate 58%), basic CPR had been performed in 32% and advanced CPR (including defibrillation) in 10%. Only 13% of the units had a written CPR policy. Units performing CPR (N = 35) were closer to an acute hospital, more likely to have short-term residents and more likely to have a CPR policy (all P < 0.05). There were no significant differences between public and private units. CONCLUSION: The widely disparate approaches to CPR in different Irish long-stay units suggest the need for national guidelines on this issue. PMID- 19190977 TI - Rapid decrease of anti-beta-glucan antibody as an indicator for early diagnosis of carinii pneumonitis and deep mycotic infections following immunosuppressive therapy in antineutrophil cytoplasmic antibody-associated vasculitis. AB - Deep mycosis (aspergillus pneumonia (AsP)) and carinii pneumonitis (PCP) are complications of immunosuppressive treatment for antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV). The objective was to clarify the clinical significance of plasma titer of antibody against beta-glucans (anti-BG antibody) as a predictor of complications such as AsP or PCP and the prognosis of patients. Enzyme-linked immunosorbent assay was used to measure the plasma titer of antibodies against beta-glucans (BG) from Candida albicans in 22 healthy subjects and 52 patients with various stages of AAV. The mean plasma titer of the anti-BG antibody was 2,677 +/- 1,686 U in healthy subjects, 691 +/- 522 U in patients with untreated active vasculitis (n = 14), and 547 +/- 416 U in patients soon after immunosuppressive treatment (n = 24). Healthy subjects had significantly higher antibody titers than the other two groups (P < 0.05). Repeated measurements over the clinical course of AAV revealed an increase during remission to 1,180 +/- 130 U (n = 11), while there was a significant rapid decrease to 369 +/- 441 U (P < 0.01) concomitantly with elevation in plasma C reactive protein and BG levels in patients with AAV that had AsP or PCP infection. Antifungal therapy resulted in a rapid rise of anti-BG antibody titer. Experiments in mice suggested that the anti-BG antibody neutralizes BG. Rapid decrease of the anti-BG antibody titer may be a useful indicator for diagnosis of the presence of AsP or PCP and for estimating the prognosis of patients with these opportunistic infections during immunosuppressive treatment of AAV. PMID- 19190979 TI - A new three-way variant t(15;22;17)(q22;q11.2;q21) in acute promyelocytic leukemia. AB - Acute promyelocytic leukemia (APL) is characterized by the t(15;17)(q22;q21), which results in the fusion of the promyelocytic leukemia (PML) gene at 15q22 with the retinoic acid alpha-receptor (RARA) at 17q21. We report the case of a 44 year-old man with APL carrying a new complex variant translocation (15;22;17). Karyotypic analysis with G-banding of bone marrow cells revealed t(15;22;17) (q22;q11.2;q21). Fluorescence in situ hybridization with a PML/RARA dual-color DNA probe showed the fusion signals. RT-PCR analysis showed long-form PML/RARA fusion transcripts. A complete remission was attained with a course of conventional chemotherapy with all-trans retinoic acid (ATRA). This is the first report of a new three-way translocation of 22q11 involvement with APL. PMID- 19190980 TI - Three challenges for the social determinants of health pursuit. PMID- 19190981 TI - On the influence of a raffle upon responses to an urban transportation survey in New York City. AB - OBJECTIVES: Determine whether raffling off prizes to increase participation in surveys triggers multiple responses from the same participants. METHODS: All staff, instructors, and students of Queens College, New York, were invited to a 5 minute online survey about commuting in winter 2007. Entry in the raffle to win one of two iPODs was not conditional upon survey completion. RESULTS: Participation was 18.7 %, and 3913 eligible responses. Of the 183 (4.7 %) participants with two answers, responses were consistent, with weighted kappas ranging between 0.48 and 1.00, and Pearson r's between 0.81 and 1.0. Kappa's were lower for opinion than for factual questions. The duplicate responses came from people with characteristics similar to the full sample. Commuting modes included car (46 %), bus but no subway (28 %), and bus and subway (21 %). The median commuting time was 90 minutes round-trip, and was greater than 120 minutes for 1/4 of the respondents. CONCLUSION: Participation was low despite the raffle. The small percentage of duplicate answers, maybe prompted by the reminding email, had a negligible impact on the results. PMID- 19190982 TI - Body shape, body size and cigarette smoking relationships. AB - OBJECTIVES: The aim of the present study was to explore whether smoking is positively related with the abdominal obesity. METHODS: Cross-sectional data was analyzed from a random sample of 1,342 subjects, stratified by their age and gender. Cigarette smoking habits were assessed and anthropometric measures obtained during the health examination. RESULTS: Statistically significant differences in waist circumference and waist-hip ratio were found among males in some of the age groups, body mass index however, was lower in older smokers. There were no differences between female smokers and non-smokers in all parameters. CONCLUSION: The results of the study do not support the hypothesis that smoking affects an abnormal fat distribution profile predominantly in the form of central adiposity as reported earlier. PMID- 19190984 TI - Explanation and falsification in phylogenetic inference: exercises in Popperian philosophy. AB - Deduction leads to causal explanation in phylogenetic inference when the evidence, the systematic character, is conceptualized as a transformation series. Also, the deductive entailment of modus tollens is satisfied when those kinds of events are operationalized as patristic difference. Arguments to the contrary are based largely on the premise that character-states are defined intensionally as objects, in terms of similarity relations. However, such relations leave biologists without epistemological access to the causal explanation and explanatory power of historical statements. Moreover, the prediction-making to which those kinds of relations are limited in practice can lead to a category error--the mental conversion of an abstraction (the classes defined in terms of similarity relations) into a thing (such as an historical individual). The latter practices and problems characterize pattern cladistics, taxa being interpreted as homeostatic property cluster natural kinds, and other instrumentalist research programs. PMID- 19190986 TI - Polyamines as a common source of hydrogen peroxide in host- and nonhost hypersensitive response during pathogen infection. AB - The hypersensitive response (HR) is a powerful resistance system that plants have developed against pathogen attack. There are two major pathways for HR induction; one is through recognition of the pathogen by a specific host protein, and is known as the host HR. The other is through common biochemical changes upon infection--the nonhost HR. We previously demonstrated that hydrogen peroxide derived from polyamine degradation by polyamine oxidase triggers the typical host HR in tobacco plants upon infection with tobacco mosaic virus. However, it remains to be determined whether or not polyamines are involved in the nonhost HR in tobacco, and in the host HR in other plant species. When tobacco plants were infected with Pseudomonas cichorii, a representative nonhost pathogen, transcripts for six genes encoding enzymes for polyamine metabolism were simultaneously induced, and polyamines were accumulated in apoplasts. Hydrogen peroxide was concomitantly produced and hypersensitive cell death occurred at infected sites. Silencing of polyamine oxidase by the virus-induced gene silencing method resulted in suppression of hydrogen peroxide production and in disappearance of visible hypersensitive cell death with an increase in bacterial growth. Our results indicated that polyamines served as the source of hydrogen peroxide during the nonhost HR in tobacco plants. Further analysis revealed that polyamines were accumulated in apoplasts of Arabidopsis thaliana infected with Pseudomonas syringae, and of rice infected with Magnaporthe grisea, both causing the typical host HR. As in tobacco, it is conceivable that the same mechanism operates for nonhost HR in these plants. Our present observations thus suggested that polyamines are commonly utilized as the source of hydrogen peroxide during host- and nonhost HRs in higher plants. PMID- 19190985 TI - Modulation of intracellular iron levels by oxidative stress implicates a novel role for iron in signal transduction. AB - Reactive oxygen species (ROS) display cytotoxicity that can be exacerbated by iron. Paradoxically, HeLa cells treated with the ROS-generators menadione and 2,3 dimethoxy-1,4-naphthoquinone display increased free labile iron. HeLa cells exposed to ROS undergo apoptosis but iron chelation limits the extent of cell death suggesting the rise in intracellular iron plays a signaling role in this pathway. This idea is supported by the fact that iron chelation also alters the pattern of ROS-induced phosphorylation of stress-activated protein kinases SAPK/JNK and p38 MAPK. Thus, ROS-induced increases in cellular free iron contribute to signaling events triggered during oxidative stress response. PMID- 19190987 TI - GASA5, a regulator of flowering time and stem growth in Arabidopsis thaliana. AB - Flowering is a critical event in the life cycle of plants and is regulated by a combination of endogenous controls and environmental cues. In the present work, we provide clear genetic evidence that GASA5, a GASA family gene in Arabidopsis (Arabidopsis thaliana), is involved in controlling flowering time and stem growth. GASA5 expression was present in all tissues of Arabidopsis plants, as detected by RT-PCR, and robust GUS staining was observed in the shoot apex of 8 day-old seedlings and inflorescence meristems during reproductive development. Phenotypic analysis showed that a GASA5 null mutant (gasa5-1) flowered earlier than wild type with a faster stem growth rate under both long-day (LD) and short day (SD) photoperiods. In contrast, transgenic plants overexpressing GASA5 demonstrated delayed flowering, with a slower stem growth rate compared to wild type plants. However, neither the GASA5 null mutants nor the GASA5 overexpressing plants revealed obvious differences in flowering time upon treatment with gibberellic acid (GA(3)), indicating that GASA5 is involved in gibberellin (GA) promoted flowering. GAI (GA INSENSITIVE), one of the five DELLAs in Arabidopsis, was more highly expressed in GASA5-overexpressing plants, but it was lower in gasa5-1. Further transcript profiling analysis suggested that GASA5 delayed flowering by enhancing FLOWERING LOCUS C (FLC) expression and repressing the expression of key flowering-time genes, FLOWERING LOCUS T (FT) and LEAFY (LFY). Our results suggest that GASA5 is a negative regulator of GA-induced flowering and stem growth. PMID- 19190989 TI - Derivation and external validation of a simple prediction model for the diagnosis of type 2 diabetes mellitus in the Brazilian urban population. AB - A risk score model was developed based in a population of 1,224 individuals from the general population without known diabetes aging 35 years or more from an urban Brazilian population sample in order to select individuals who should be screened in subsequent testing and improve the efficacy of public health assurance. External validation was performed in a second, independent, population from a different city ascertained through a similar epidemiological protocol. The risk score was developed by multiple logistic regression and model performance and cutoff values were derived from a receiver operating characteristic curve. Model's capacity of predicting fasting blood glucose levels was tested analyzing data from a 5-year follow-up protocol conducted in the general population. Items independently and significantly associated with diabetes were age, BMI and known hypertension. Sensitivity, specificity and proportion of further testing necessary for the best cutoff value were 75.9, 66.9 and 37.2%, respectively. External validation confirmed the model's adequacy (AUC equal to 0.72). Finally, model score was also capable of predicting fasting blood glucose progression in non-diabetic individuals in a 5-year follow-up period. In conclusion, this simple diabetes risk score was able to identify individuals with an increased likelihood of having diabetes and it can be used to stratify subpopulations in which performing of subsequent tests is necessary and probably cost-effective. PMID- 19190988 TI - Chronic exposure of arsenic via drinking water and its adverse health impacts on humans. AB - Worldwide chronic arsenic (As) toxicity has become a human health threat. Arsenic exposure to humans mainly occurs from the ingestion of As contaminated water and food. This communication presents a review of current research conducted on the adverse health effects on humans exposed to As-contaminated water. Chronic exposure of As via drinking water causes various types of skin lesions such as melanosis, leucomelanosis, and keratosis. Other manifestations include neurological effects, obstetric problems, high blood pressure, diabetes mellitus, diseases of the respiratory system and of blood vessels including cardiovascular, and cancers typically involving the skin, lung, and bladder. The skin seems to be quite susceptible to the effects of As. Arsenic-induced skin lesions seem to be the most common and initial symptoms of arsenicosis. More systematic studies are needed to determine the link between As exposure and its related cancer and noncancer end points. PMID- 19190991 TI - Assessment of contaminant impacts in a semi-enclosed estuary (Amvrakikos Gulf, NW Greece): bioenergetics and biochemical biomarkers in mussels. AB - A combination of bioenergetics and biochemical biomarkers in mussels was applied to assess possible pollution impacts in a protected semi-enclosed estuary (Amvrakikos Gulf, NW Greece) that receives pesticide discharges through riverine transport. Scope for growth, a physiological condition index representing the energy budget of the organism, was applied to detect general stress effects on the health status of mussels. The low energy budgets of mussels revealed stress conditions and provided early warning signals of possible consequences at higher levels of biological organization. Biochemical markers of exposure confirmed a risk of pesticide contamination. Decreased acetylcholinesterase activities indicated exposure to organophosphate and carbamate pesticides. Responses of the antioxidant enzyme glutathione peroxidase suggested the presence of contaminants capable of reactive oxygen species production that could be related to organochlorine pesticide contamination in the area. On the other hand, metallothionein levels implied low metal contamination. PMID- 19190992 TI - Assessing forest canopy closure in a geospatial medium to address management concerns for tropical islands--Southeast Asia. AB - The present study outlines an approach to classify forest density and to estimate canopy closure of the forest of the Andaman and Nicobar archipelago. The vector layers generated for the study area using satellite data was validated with the field knowledge of the surveyed ground control points. The methodology adopted in this present analysis is three-tiered. First, the density stratification into five zones using visual interpretation for the complete archipelago. In the second step, we identified two island groups from the Andaman to investigate and compare the forest strata density. The third and final step involved more of a localised phytosociological module that focused on the North Andaman Islands. The results based on the analysis of the high-resolution satellite data show that more than 75% of the mangroves are under high- to very high-density canopy class. The framework developed would serve as a significant measure to forest health and evaluate management concerns whilst addressing issues such as gap identification, conservation prioritisation and disaster management--principally to the post tsunami assessment and analysis. PMID- 19190990 TI - Umbrea, a chromo shadow domain protein in Drosophila melanogaster heterochromatin, interacts with Hip, HP1 and HOAP. AB - Drosophila melanogaster HP1-interacting protein (Hip) is a partner of heterochromatin protein 1 (HP1) and is involved in transcriptional epigenetic gene silencing and the formation of heterochromatin. Recently, it has been shown that HP1 interacts with the telomere capping factor HP1/ORC (origin recognition complex)-associated protein (HOAP). Telomeres, complexes of DNA and proteins at the end of linear chromosomes, have been recognized to protect chromosome ends from degradation and fusion events. Both proteins are located at telomeres and prevent telomere fusions. Here, we report the identification and characterization of the Hip-interacting protein Umbrea. We found that Umbrea interacts directly with Hip, HP1 and HOAP in vitro. Umbrea, Hip and HP1 are partners in a protein complex in vivo and completely co-localize in the pericentric heterochromatin and at telomeres. Using a Gal4-induced RNA interference system, we found that after depletion of Umbrea in salivary gland polytene chromosomes, they exhibit multiple telomeric fusions. Taken together, these results suggest that Umbrea cooperates with Hip, HP1 and HOAP and plays a functional role in mediating normal telomere behaviour in Drosophila. PMID- 19190993 TI - Environmental applications of ICP-MS for simultaneous determination of trace elements and statistical data analysis. AB - Water samples collected from two lakes separated from each other by the cascade of four small lowland reservoirs were subjected to analyses in order to compare contamination levels in these ecosystems. Inductively coupled plasma mass spectrometry was used to determine eight elements: Al, As, Cd, Cr, Li, Mn, Sb, and Zn. Statistical methods were applied in order to assess the degree of self purification of water, during its flow from Swarzedzkie Lake to Malta Lake. Obtained distinctly, lower concentrations of all studied elements in Malta Lake than in Swarzedzkie Lake indicated the role of self-purification processes, which occurred in the distance between these lakes. PMID- 19190994 TI - QSAR modelling of carcinogenicity by balance of correlations. AB - Optimal descriptors based on the simplified molecular input line entry system (SMILES) have been utilized in modeling of carcinogenicity. Carcinogenicity of 401 compounds has been modeled by means of balance of correlations for the training (n = 170) and calibration (n = 170) sets. The obtained models were evaluated with an external test set (n = 61). Comparison of models based on the balance of correlations and models which were obtained on the basis of the total training set (i.e., both training and calibration sets as the united training set) has shown that the balance of correlations improves the statistical quality for the external test set. PMID- 19190995 TI - Effectiveness of a community-based strategic anthelmintic treatment programme in the control of gastrointestinal nematodes and Fasciola gigantica in cattle in Kilolo district, Tanzania. AB - A longitudinal demonstration project was carried out in 2005/2006 for thirteen months to determine the effectiveness of a strategic community-based worm control (CBWC) programme in the control of gastrointestinal (GI) nematodes and Fasciola gigantica in cattle. Two villages namely, a CBWC village (Ilula-Masukanzi) and a village without CBWC (Kilolo-Luganga) were randomly selected in Kilolo district, Iringa region of Tanzania. Animals in both villages were under the traditional management system and used the village communal grazing area. At the village with CBWC, 60 zebu cattle (6-18 months old) from 10 farms in the village were selected and ear tagged for monthly sampling and weghing. A strategic CBWC programme was instituted whereby all animals in the village (tagged and non-tagged) were treated with albendazole 10% drench at 10 mg/kg four times a year e.g. middle of the rainy season (February), end of the rainy season (May), middle of the dry season (September) and late dry/early rainy season (December). In the village without CBWC, 68 zebu cattle (6-18 months old) from 11 farms in the village were selected and tagged for monthly sampling and weighing. Farmers in the village without CBWC were allowed to continue with their normal management practices including anthelmintic treatments. Faecal and blood samples from tagged animals and pasture samples from communal grazing areas were collected on monthly basis for 13 months. Results showed that strategic CBWC programme was highly effective in reducing faecal egg counts (FEC) and the proportion of animals passing Fasciola eggs in faeces. The strategic CBWC significantly reduced FEC of animals by an average of 60% compared to animals at the village without CBWC (P < 0.01). Four strategic CBWC treatments per year significantly reduced the proportion of animals passing Fasciola eggs in faeces by an average of 82.5% compared to animals at the village without CBWC (P < 0.05). From commencement to the end of the trial, animals at the village with CBWC significantly outgained animals at the village without CBWC by an average of 13.9 kg per year (P < .05). It is concluded that strategic CBWC programme was highly effective in reducing FEC, reducing the proportion of animals passing Fasciola eggs in faeces and improved weight gain in animals. PMID- 19190996 TI - Magnetic resonance imaging of the normal bovine digit. AB - The purpose of this study was defining the normal structures of the digits and hoof in Holstein dairy cattle using Magnetic Resonance Image (MRI). Transverse, Sagital and Dorsoplantar MRI images of three isolated cattle cadaver digits were obtained using Gyroscan T5-NT a magnet of 0.5 Tesla and T1 Weighted sequence. The MRI images were compared to corresponding frozen cross-sections and dissect specimens of the cadaver digits. Relevant anatomical structures were identified and labeled at each level. The MRI images provided anatomical detail of the digits and hoof in Holstein dairy cattle. Transversal images provided excellent depiction of anatomical structures when compared to corresponding frozen cross sections. The information presented in this paper would serve as an initial reference to the evaluation of MRI images of the digits and hoof in Holstein dairy cattle, that can be used by radiologist, clinicians, surgeon or for research propose in bovine lameness. PMID- 19190997 TI - Effects of propofol on pro-inflammatory cytokines and nuclear factor kappaB during polymicrobial sepsis in rats. AB - Nuclear factor kappa B (NF-kappaB) plays a central role in regulating the transcription of several genes associated with sepsis/septic shock. Therefore, the author investigated the effects of propofol on the plasma tumor necrosis factor alpha and interleukin 6 (TNF-alpha and IL-6) levels and NF-kappaB activation during polymicrobial sepsis in rats. Male Sprague-Dawlay rats were subjected to cecal ligation and puncture (CLP, a model of polymicrobial sepsis) or sham operation. The animals were randomly assigned into four equal groups (n = 10): sham CLP group, CLP group, PPF (propofol) I group and PPF II group. Thirty minutes before CLP, propofol (5 and 10 mg kg(-1) h(-1), respectively) was infused continuously through the left femoral vein cannula in PPF I group or PPF II group, CLP group and sham CLP group receiving 0.9% saline only at the rates of 5 ml kg(-1) h(-1). The right femoral artery was cannulated to monitor mean arterial pressure (MAP) and heart rates (HR). CLP produced progressive hypotension and a first increase followed by a decrease in HR. The plasma TNF-alpha and IL-6 levels and the hepatic NF-kappaB activation significantly increased after CLP alone. Compared with CLP group, propofol treatment reversed hypotension, slightly steadied heartbeats, and decreased the plasma TNF-alpha and IL-6 levels, and significantly suppressed NF-kappaB activation. Propofol has inhibited the hepatic NF-kappaB activation and the pro-inflammatory cytokine response during polymicrobial sepsis in rats. PMID- 19190999 TI - Disability meanings according to patients and clinicians: imagined recovery choice pathways. AB - OBJECTIVE: The purpose of this study was to explore how the meaning of disability varies between patients with acute-onset activity limitations and clinicians, and between males and females. METHODS: Seventy-nine patients undergoing inpatient rehabilitation and 93 practicing rehabilitation clinicians in the USA developed personal recovery choice pathways through recovery preference exploration (RPE). Imagining complete dependence in 18 activities as diverse as eating and expression, each individual determined an optimal sequence of recovery. This sequence was used to determine the relative value of each activity compared with the other 17. Three comparisons were made by calculating the mean absolute difference (MAD) in median utilities, including patients versus clinicians, male versus female patients, and male versus female clinicians. The MAD shows the relative magnitude of disparity between each pair. RESULTS: The MAD value between patients and clinicians was 3.4 times larger and 4.8 times larger than the MAD values between male and female patients and male and female clinicians, respectively. CONCLUSIONS: The much larger difference in recovery preferences between patients and clinicians compared with differences between genders suggests that life contexts of being a patient with disabilities versus a clinician are more potent determinants of activity limitation perspectives than being a man or woman. PMID- 19190998 TI - Delivery of HSF1(+) protein using HIV-1 TAT protein transduction domain. AB - HSF1 is the major transcription factor of HSPs (heat shock proteins) in response to various stresses. Wild type HSF1 (heat shock transcriptional factor 1) is normally inactive, while a constitutively active form of HSF1 (HSF1(+)) can activate downstream HSP expression in the absence of stresses. Here we generated the eukaryotic vectors that expresses HSF1(+) fusion proteins, and found that HSF1(+)-TAT fusion protein was expressed and activated HSP expression. TAT, as a trans-acting factor of HIV-1, has been demonstrated to deliver functional cargo protein into living cells. HSF1(+)-TAT fusion protein was expressed in E. coli, purified, incubated with A549 cells for 8 h, Western blot analysis and luciferase reporter assay showed that HSF1(+) fusion protein was delivered into A549 cells successfully, and the accumulation of HSF1(+)-TAT fusion protein in A549 cells up regulated HSP70 expression. PMID- 19191000 TI - LTD windows of the STDP learning rule and synaptic connections having a large transmission delay enable robust sequence learning amid background noise. AB - Spike-timing-dependent synaptic plasticity (STDP) is a simple and effective learning rule for sequence learning. However, synapses being subject to STDP rules are readily influenced in noisy circumstances because synaptic conductances are modified by pre- and postsynaptic spikes elicited within a few tens of milliseconds, regardless of whether those spikes convey information or not. Noisy firing existing everywhere in the brain may induce irrelevant enhancement of synaptic connections through STDP rules and would result in uncertain memory encoding and obscure memory patterns. We will here show that the LTD windows of the STDP rules enable robust sequence learning amid background noise in cooperation with a large signal transmission delay between neurons and a theta rhythm, using a network model of the entorhinal cortex layer II with entorhinal hippocampal loop connections. The important element of the present model for robust sequence learning amid background noise is the symmetric STDP rule having LTD windows on both sides of the LTP window, in addition to the loop connections having a large signal transmission delay and the theta rhythm pacing activities of stellate cells. Above all, the LTD window in the range of positive spike timing is important to prevent influences of noise with the progress of sequence learning. PMID- 19191001 TI - Deep analysis of perception through dynamic structures that emerge in cortical activity from self-regulated noise. AB - The statistical properties of the spontaneous background electrocorticogram (ECoG) were modeled, starting with random numbers, constraining the distributions, and identifying characteristic deviations from randomness in ECoG from subjects at rest and during intentional behaviors. The ECoG had been recorded through 8 x 8 arrays of 64 electrodes, from the surfaces of auditory, visual, or somatic cortices of 9 rabbits, and from the inferotemporal cortex of a human subject. Power spectral densities (PSD) in coordinates of log(10) power versus log(10) frequency of ECoG from subjects at rest usually conformed to noise in power-law distributions in a continuum. PSD of ECoG from active subjects usually deviated from noise in having peaks in log(10) power above the power-law line in various frequency bands. The analytic signals from the Hilbert transform after band pass filtering in the beta and gamma ranges revealed beats from interference among distributed frequencies in band pass filtered noise called Rayleigh noise. The beats were displayed as repetitive down spikes in log(10) analytic power. Repetition rates were proportional to filter bandwidths for all center frequencies. Resting ECoG often gave histograms of the magnitudes and intervals of down spikes that conformed to noise. Histograms from active ECoG often deviated from noise in Rayleigh distributions of down spike intervals by giving what are called Rice (Mathematical analysis of random noise-and appendixes technical publications monograph B-1589. Bell Telephone Labs Inc., New York, 1950) distributions. Adding power to noise as signals at single frequencies simulated those deviations. The beats in dynamic theory are deemed essential for perception, by gating beta and gamma bursts at theta rates through enhancement of the cortical signal-to-noise ratio in exceptionally deep down spikes called null spikes. PMID- 19191002 TI - Underrepresentation in the real world. PMID- 19191003 TI - On the ethics of using citation indices in evaluations. PMID- 19191004 TI - Brain dysfunction in phenylketonuria: is phenylalanine toxicity the only possible cause? AB - In phenylketonuria, mental retardation is prevented by a diet that severely restricts natural protein and is supplemented with a phenylalanine-free amino acid mixture. The result is an almost normal outcome, although some neuropsychological disturbances remain. The pathology underlying cognitive dysfunction in phenylketonuria is unknown, although it is clear that the high plasma concentrations of phenylalanine influence the blood-brain barrier transport of large neutral amino acids. The high plasma phenylalanine concentrations increase phenylalanine entry into brain and restrict the entry of other large neutral amino acids. In the literature, emphasis has been on high brain phenylalanine as the pathological substrate that causes mental retardation. Phenylalanine was found to interfere with different cerebral enzyme systems. However, apart from the neurotoxicity of phenylalanine, a deficiency of the other large neutral amino acids in brain may also be an important factor affecting cognitive function in phenylketonuria. Cerebral protein synthesis was found to be disturbed in a mouse model of phenylketonuria and could be caused by shortage of large neutral amino acids instead of high levels of phenylalanine. Therefore, in this review we emphasize the possibility of a different idea about the pathogenesis of mental dysfunction in phenylketonuria patients and the aim of treatment strategies. The aim of treatment in phenylketonuria might be to normalize cerebral concentrations of all large neutral amino acids rather than prevent high cerebral phenylalanine concentrations alone. In-depth studies are necessary to investigate the role of large neutral amino acid deficiencies in brain. PMID- 19191005 TI - PKU-what is daily practice in various centres in Europe? Data from a questionnaire by the scientific advisory committee of the European Society of Phenylketonuria and Allied Disorders. AB - BACKGROUND: Since the start of the European Society of Phenylketonuria and Allied Disorders Treated as Phenylketonuria (ESPKU) in 1987, an increasing number of parental organizations of member countries have joined. Treatment varies widely within Europe. A survey among professionals was done to determine goals and practice. METHOD: In 2005, a questionnaire was sent to professionals of member countries, addressing diagnostic and treatment procedures, numbers of patients necessary for a PKU centre, guidelines followed, numbers of patients treated and professionals involved in care, target phenylalanine concentrations, amount of protein prescribed, frequency of monitoring and clinical visits, need for follow up of various clinical and biochemical data, the importance of various abnormalities, and definition of (non)compliance. RESULTS: Seventeen centres of 12 countries answered. Professionals of 13 countries could not be reached or did not respond. Differences in care were observed in many issues of care including target phenylalanine concentrations. Only few issues had general consensus. CONCLUSION: Not all countries were really active at ESPKU level. In the active countries, a professional could not always be contacted. Responses show that PKU care varies largely between European countries. Notwithstanding the large diversity on many issues of day-to-day care and therapeutic targets, results showed increasing consensus on some issues. The most important outcome of this questionnaire might be that the Scientific Advisory Committee of the ESPKU initiated meetings for professionals of different backgrounds taking care of PKU patients besides the already existing programme for parents, patients and delegates. Discussion among these professionals may improve quality of care. PMID- 19191006 TI - Bloodspot acylcarnitine and amino acid analysis in cord blood samples: efficacy and reference data from a large cohort study. AB - BACKGROUND: In order to test the feasibility of cord blood screening for inherited metabolic disease, a two-year cohort study of births in six obstetric units from five towns in the north of England was undertaken. These towns have a high prevalence of consanguineous marriages, largely among the immigrant Asian community. The purpose of the study was to determine whether early detection of metabolic disease was possible and whether early intervention would improve prognosis. METHODS: Following parental consent, cord blood samples were collected at birth and analysed for acylcarnitine and amino acid profiles by tandem mass spectrometry in one of two laboratories. One laboratory used butylated derivatives, the other used underivatized samples. The same laboratories performed routine blood spot neonatal screening at 5-7 days of age on these babies. Patients with positive results were investigated and treated by a metabolic paediatrician as soon as possible. RESULTS: 24,983 births were examined. 12,952 samples were analysed as butyl derivatives, 12,031 samples were analysed underivatized. The following disorders were detected: medium-chain acyl CoA dehydrogenase (MCAD) deficiency (1 case), 3-methylcrotonyl-CoA carboxylase (MCC) deficiency (2 cases), maternal carnitine transporter defect (2 cases), maternal MCC (1 case). The following disorders were diagnosed subsequently but were not detected by the cord blood screening: phenylketonuria (PKU) (1 case), maple syrup urine disease (MSUD) (2 cases), argininosuccinic aciduria (1 case), methylmalonic acidaemia (MMA) (1 case), glutaric aciduria type 2 (1 case), MCAD deficiency (2 cases), 3-hydroxy-3-methylglutaryl-CoA lyase deficiency (1 case). Comprehensive reference data for all analytes by both methods were obtained. CONCLUSIONS: Cord blood testing is of limited value in detecting inherited metabolic disease. The metabolites associated with most disorders examined were not elevated in cord blood. Some maternal disorders, carnitine transporter defect and 3-methlycrotonyl-CoA carboxylase deficiency, are detected. These remain of uncertain clinical significance. Comprehensive reference data have been obtained that will facilitate future interpretation of studies in cord blood. PMID- 19191007 TI - Role of heat shock protein 27 phosphorylation in migration of vascular smooth muscle cells. AB - OBJECTIVE: The aim of the present study was to investigate the role of heat shock protein 27 (HSP27) phosphorylation in the migration of vascular smooth muscle cells (VSMCs) induced by angiotensin II (AngII) and platelet derived growth factor-BB (PDGF-BB). METHODS: The activity of HSP27 was evaluated by Western blot with specific phospho-HSP27 antibody. F-actin polymerization was detected by FITC Phalloidine staining using confocal microscopy. Modified Boyden chamber technique was employed for VSMCs migration assessment. RESULTS: The phosphorylation of HSP27 was induced by AngII and PDGF-BB in a time- and concentration-dependent manner in VSMCs, which was significantly blocked by the HSP inhibitor Quercetin in a concentration-dependent manner. Reorganization of actin stimulated by AngII and PDGF-BB was markedly inhibited by pretreatment with 100 micromol/l Quercetin. The migration of VSMCs induced by AngII and PDGF-BB was partially inhibited by Quercetin with peak inhibition concentration at 100 micromol/l. CONCLUSIONS: HSP27 phosphorylation plays an important role in mediating the rearrangement of F actin and migration of VSMCs induced by AngII and PDGF-BB. HSP27 may be a potential target for the interventional treatment of pathological process related to cell migration. PMID- 19191008 TI - PI-3 K/AKT and ERK signaling pathways mediate leptin-induced inhibition of PPARgamma gene expression in primary rat hepatic stellate cells. AB - Compelling evidence indicates the pro-fibrogenic action of leptin in liver. Peroxisome proliferator-activated receptor-gamma (PPARgamma) can reverse hepatic stellate cell (HSC) activation and maintain HSC quiescence. HSC activation, a key step in the development of liver fibrosis, is coupled with the up-expression of leptin and the dramatic down-expression of PPARgamma. The present study is aimed to assess the effect of leptin on PPARgamma gene expression in primary cultured rat HSCs and investigate the related mechanisms by using Western blotting analysis, real-time PCR, transient transfection approach, and cell growth analysis. The results suggest that leptin negatively regulates PPARgamma gene expression at mRNA level, protein level and PPARgamma gene promoter activity level in HSCs. The inhibitory effect of leptin on PPARgamma gene expression contributes to cell growth of activated HSCs in vitro. Phosphatidylinositol 3 kinase/AKT (PI-3 K/AKT) and extracellular signal-regulated kinase (ERK) signaling pathways mediate the leptin-induced inhibition of PPARgamma gene expression. In summary, these findings suggest that leptin down-regulates PPARgamma gene expression through activation of PI-3 K/AKT or ERK signaling pathway in primary cultured rat HSCs. Our results might provide novel insights into the mechanisms for the pro-fibrogenic action of leptin in liver. PMID- 19191009 TI - Isoenzyme-specific up-regulation of glutathione transferase and aldo-keto reductase mRNA expression by dietary quercetin in rat liver. AB - The impact of quercetin on the mRNA expression of hepatic enzymes involved in drug metabolism was evaluated with a DNA microarray and real-time PCR. Male Sprague-Dawley rats were fed an experimental diet containing either 0, 2.5, 5, 10, or 20 g/kg of quercetin for 15 days. The DNA microarray analysis of the gene expression profile in pooled RNA samples from rats fed diets containing 0, 5, and 20 g/kg of quercetin revealed genes of some isoenzymes of glutathione transferase (Gst) and aldo-keto reductase (Akr) to be activated by this flavonoid. Real-time PCR conducted with RNA samples from individual rats fed varying amounts of quercetin together with the microarray analysis showed that quercetin caused marked dose-dependent increases in the mRNA expression of Gsta3, Gstp1, and Gstt3. Some moderate increases were also noted in the mRNA expression of isoenzymes belonging to the Gstm class. Quercetin also dose-dependently increased the mRNA expression of Akr1b8 and Akr7a3. However, it did not affect the parameters of the other Gst and Akr isoenzymes. It is apparent that quercetin increases the mRNA expression of Gst and Akr involved in drug metabolism in an isoenzyme-specific manner. Inasmuch as Gst and Akr isoenzymes up-regulated in their gene expression are involved in the prevention and attenuation of cancer development, this consequence may account for the chemopreventive propensity of quercetin. PMID- 19191010 TI - Molecular mechanism of curcumin induced cytotoxicity in human cervical carcinoma cells. AB - Cervical cancer is the most common cancer in Indian females and is associated with infection with high risk Human papillomaviruses (HPVs). Curcumin (Diferuloyl methane), a chemopreventive agent, is a natural compound extracted from Curcuma longa that allows suppression of carcinogenesis. The objective of this study was to identify the molecular mechanism of curcumin induced apoptosis in HPV positive cervical cancer HeLa, SiHa and Ca Ski cells. Curcumin causes distinct inhibition of human telomerase reverse transcriptase (hTERT) the catalytic core of telomerase thereby reducing proliferation of cancer cells. Curcumin mediated apoptosis in these cells appears to be due to upregulation of proapoptotic Bax, AIF, release of cytochrome c and down regulation of antiapoptotic Bcl-2, Bcl-XL in HeLa and SiHa. This was accompanied by an increase in caspase-3 and -9 activity, suggesting the role of mitochondria in curcumin mediated apoptotic cell death. Curcumin acts as an anti-inflammatory and anti-proliferative agent by causing down regulation of COX-2, iNOS and cyclin D1 in all the three cell lines but to different extent. PMID- 19191011 TI - Highly efficient transfection of rat cortical neurons using carbosilane dendrimers unveils a neuroprotective role for HIF-1alpha in early chemical hypoxia-mediated neurotoxicity. AB - PURPOSE: To study the effect of a non-viral vector (carbosilane dendrimer) to efficiently deliver small interfering RNA to postmitotic neurons to study the function of hypoxia-inducible factor-1alpha (HIF1-alpha) during chemical hypoxia mediated neurotoxicity. METHODS: Chemical hypoxia was induced in primary rat cortical neurons by exposure to CoCl(2). HIF1-alpha levels were determined by Western Blot and toxicity was evaluated by both MTT and LDH assays. Neurons were incubated with dendriplexes containing anti-HIF1-alpha siRNA and both uptake and HIF1-alpha knockdown efficiency were evaluated. RESULTS: We report that a non viral vector (carbosilane dendrimer) can deliver specific siRNA to neurons and selectively block HIF1-alpha synthesis with similar efficiency to that achieved by viral vectors. Using this method, we have found that this transcription factor plays a neuroprotective role during the early phase of chemical hypoxia-mediated neurotoxicity. CONCLUSION: This work represents a proof-of-concept for the use of carbosilane dendrimers to deliver specific siRNA to postmitotic neurons to block selected protein synthesis. This indicates that this type of vector is a good alternative to viral vectors to achieve very high transfection levels in neurons. This also suggests that carbosilane dendrimers might be very useful for gene therapy. PMID- 19191013 TI - A comparative study into two dual fluorescent mechanisms via positional isomers of N-hydroxyarene-1,8-naphthalimides. AB - Three isomers of hydroxy substituted N-aryl-1, 8-naphthalimides based on N-aryl naphthalic anhydride fluorophore have been synthesized. The decrease in fluorescence intensity from ortho to para substitution of hydroxy group on N-aryl reveals that para substituted isomer undergoes ESEC (Excited State with Extended Conjugation) mechanism which is proved by low quantum yield and appearance of dual emission. The ortho isomer, however, has high quantum yield and no tautomer emission, indicating ESIPT (Excited State Intramolecular Proton Transfer) mechanism is not operating. Similarly, all these isomers show strong fluorescence quenching in presence of strong H-bonding solvents like DMSO and pyridine, but there was neither the shift of emission bands nor the appearance of new bands for proton transfer to these solvents. Thus, it also indicates the absence of excited state proton transfer mechanism. Both the ortho isomer, and to a greater degree the meta isomer, showed larger quenching constants (Kapp) with pyridine than DMSO. This trend opposes the hydrogen-bond affinity for these solvents with phenol and points to a 2-point recognition interaction. In addition, a naphthalimide derivative using 2-aminoimidazole was prepared and examined for optimal positioning of a six-membered ring hydrogen bond pattern. No dual fluorescence was observed for this compound either. PMID- 19191012 TI - Design of a multifunctional PLGA nanoparticulate drug delivery system: evaluation of its physicochemical properties and anticancer activity to malignant cancer cells. AB - PURPOSE: Several individual approaches were combined to fabricate a novel nanoparticulate drug delivery system to achieve targeting and anticancer effects in various malignant cancer cells. METHODS: Doxorubicin was conjugated to Poly(lactic-co-glycolic acid) (PLGA), which was formulated into nanoparticle via solvent-diffusion method. The surface of the nanoparticles was subsequently linked with Poly(ethylene glycol) (PEG) and Arg-Gly-Asp (RGD) peptide to realize both passive and active targeting functions. The multifunctional nanoparticles were then tested against several malignant cancer cell lines. RESULTS: The conjugation increased loading efficiency of doxorubicin to PLGA nanoparticles (the encapsulation efficiency was over 85%) and alleviated the drug burst release effect substantially. The drug was released from the polymeric matrix in a sustained release manner over a period of 12 days. The resultant nanoparticles were spherically uniform and well-dispersed. The nanoparticle targeting ability was proven through strong affinity to various integrin-expressing cancer cells, and much less affinity to the low integrin expression cancer cells. The nanoparticles also showed high efficacy in inducing apoptosis in specific malignant cancer cell. CONCLUSION: The developed multifunctional nanoparticles hold potential to treat malignant integrin-expressing cancers. PMID- 19191014 TI - The theoretical three-dimensional structure of Bacillus thuringiensis Cry5Aa and its biological implications. AB - Cry5Aa is a crystal protein produced by Bacillus thuringiensis serovar. damstadiensis during its stationary phase, this delta-endotoxin is active against nematodes and has great potential for nematodes control. The theoretical model of the three-dimensional structure of Cry5Aa was predicted by homology modeling on the structures of the Cry1Aa which is specific to Lepidopteran insects. The structure of the Cry5Aa resembles previously reported Cry toxin structures but shows the following distinctions. Cry5Aa has a long insertion in alpha2 of domain I. Some loops in the domain II and III of Cry5Aa are exposed to the solvent. In this work we give a brief description of our model and hypothesize the residues of the Cry5Aa that could be important in receptor recognition and pore formation. This model will be helpful for the design of mutagenesis experiments aimed to the improvement of toxicity, and lead to a deep understanding of the mechanism of action of nematicidal toxins. PMID- 19191015 TI - Purinergic signalling in rat GFSHR-17 granulosa cells: an in vitro model of granulosa cells in maturing follicles. AB - Purinergic signalling in rat GFSHR-17 granulosa cells was characterised by Ca(2+) imaging and perforated patch-clamp. We observed a resting intracellular Ca(2+) concentration ([Ca(2+)](i)) of 100 nM and a membrane potential of -40 mV. This was consistent with high K(+)- and Cl(-) permeability and a high intracellular Cl(-) concentration of 40 mM. Application of ATP for 5-15 s every 3 min induced repeated [Ca(2+)](i) increases and a 30 mV hyperpolarization. The phospholipase C inhibitor U73122 or the IP(3)-receptor antagonist 2-aminoethoethyl diphenyl borate suppressed ATP responses. Further biochemical and pharmacological experiments revealed that ATP responses were related to stimulation of P2Y(2) and P2Y(4) receptors and that the [Ca(2+)](i) increase was a prerequisite for hyperpolarization. Inhibitors of Ca(2+)-activated channels or K(+) channels did not affect the ATP-evoked responses. Conversely, inhibitors of Cl(-) channels hyperpolarized cells to -70 mV and suppressed further ATP-evoked hyperpolarization. We propose that P2Y(2) and P2Y(4) receptors in granulosa cells modulate Cl(-) permeability by regulating Ca(2+)-release. PMID- 19191016 TI - Multi-informant ratings of psychiatric symptom severity in children with autism spectrum disorders: the importance of environmental context. AB - The present study examines co-occurring psychiatric syndromes in a well characterized sample of youths with autism spectrum disorders (ASD; n = 177) and their siblings (n = 148), reported independently by parents and teachers. In ASD, parents reported substantial comorbidity with affective (26%), anxiety (25%), attentional (25%), conduct (16%), oppositional (15%), and somatic problems (6%). Teachers reported a much lower prevalence. Autistic severity scores for children with ASD exhibited moderate correlations with general psychopathology within- but not across-informants, whereas, sibling correlations were significant both within and across-informants. Results support the role of environmental context in psychiatric symptom expression in children affected by autism and suggest that informant discrepancies may more provide critical cues for these children via specific environmental modifications. PMID- 19191017 TI - The use of structural analysis to develop antecedent-based interventions for students with autism. AB - Evidence continues to maintain that the use of antecedent variables (i.e., instructional practices, and environmental characteristics) increase prosocial and adaptive behaviors of students with disabilities (e.g., Kern et al. in J Appl Behav Anal 27(1):7-19, 1994; Stichter et al. in Behav Disord 30:401-418, 2005). This study extends the literature by systematically utilizing practitioner implemented structural analyzes within school settings to determine antecedent variables affecting the prosocial behavior of students with autism. Optimal antecedents were combined into intervention packages and assessed utilizing a multiple baseline design across settings. All three students demonstrated improvement across all three settings. Rates of engagement and social interaction were obtained from classroom peers to serve as benchmark data. Findings indicate that practitioners can implement structural analyzes and design corresponding interventions for students with ASD within educational settings. PMID- 19191018 TI - Cognitive schemas associated with negative sexual events: a comparison of men and women with and without sexual dysfunction. AB - The association between cognitive schemas activated during sexual events and sexual functioning was studied. A total of 376 participants (47 women and 49 men with a DSM-IV diagnosis of sexual dysfunction and 160 women and 120 men from a control group) answered the Questionnaire of Cognitive Schema Activation in Sexual Context, the International Index of Erectile Function, and the Female Sexual Function Index. Results showed that participants with sexual dysfunction activated significantly more negative schemas when exposed to sexually unsuccessful situations compared to sexually healthy individuals (after controlling for the frequency of negative sexual events experienced by both groups). Most men and women with sexual difficulties interpreted negative sexual events according to an incompetence self-schema (I'm powerless, I'm incompetent, I'm a failure). These findings were congruent with recent research indicating the tendency shown by individuals with sexual dysfunction to give attributions of an internal, stable, and global nature to negative sexual experiences. Overall, results suggest specific faulty cognitive constructions underlying sexual dysfunctions and encourage the development of models and treatment approaches based on cognitive theory. PMID- 19191019 TI - Cloning and characterization of the promoter of the human AHI1 gene. AB - The AHI1 gene is involved in axons crossing the midline during development, and its mutation can cause Joubert syndrome and other related neurological system diseases. Its transcriptional regulation mechanism is still unknown. Here we cloned and characterized the upstream 5' region of the gene. It was shown that 1.3 kb upstream of ATG had the transcription activity. Progressive deletion analysis was done, and all the constructs were transfected into COS7 cells. The promoter activity was determined by fluorescence microscopy, flow cytometry, and reverse transcription polymerase chain reaction. Our results showed that all the constructs had transcription activity. The highest promoter activity was 30 bp upstream of ATG, which included a transcription factor-binding site for Oct-1. Our research provides an important basis for further investigations on the transcriptional regulation of the human AHI1 gene. PMID- 19191020 TI - Isolation and characteristics of the CN gene, a tobacco mosaic virus resistance N gene homolog, from tobacco. AB - Nicotiana rustica L. HZNH, a native Chinese tobacco germplasm, displays a hypersensitive response (HR) and systemic acquired resistance following infection with tobacco mosaic virus (TMV). A resistance gene, CN, cloned from HZNH plants, was homologous to the N and NH genes identified in other Nicotiana species. The CN coding region (3423 bp) shares 93.63% and 86.50% nucleotide identity with N and NH, respectively. Whereas the five CN exon sequences are highly homologous with those of N and NH, the four introns differ significantly in length and sequence. Sequence analysis revealed that CN belongs to the TIR/NBS/LRR gene class. Expression of CN was up-regulated after TMV infection and was temperature sensitive. Organ-specific expression analysis suggested that CN transcripts accumulated at high levels in leaves, low levels in stems, and minimal levels in roots. When CN was inserted into TMV-susceptible N. tabacum cv. K326 plants by Agrobacterium-mediated transformation, the transgenic plants displayed HR and systemic HR due to uninhibited movement of the virus. PMID- 19191021 TI - Correlates of existential well-being and their association with health-related quality of life in breast cancer survivors compared with the general population. AB - The aim was to evaluate the correlates of existential well-being (EWB) and investigate the relationship between EWB and health-related quality of life (HRQOL) according to the McGill Quality of Life Questionnaire (MQOL) in breast cancer survivors (BCS) and in the general population. BCS (N = 1,933) recruited from five large hospitals completed a mailed survey, which included the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire Core 30 (QLQ-C30), Quality of Life Questionnaire Breast Cancer Module (QLQ-BR23) and the MQOL. Reference data were derived from a representative sample of the Korean adult population composed of 500 women selected by a nationwide random route technique. Multiple logistic regression analysis showed that members of the general population with a lower educational status were more likely to report poor EWB. However, among BCS, correlates that were associated with poor EWB included lower monthly income, lower educational status, unemployment and comorbidities. Almost all factors listed on the HRQOL questionnaire were correlated with EWB in BCS as well as in the general population (P < 0.001). Factors listed on the QLQ-C30 that were correlated to a clinically meaningful extent with EWB in the general population included physical functioning, nausea and vomiting, and appetite loss, whereas among BCS, insomnia and distress over hair loss were meaningfully correlated with EWB. Compared with general population, BCS showed different EWB correlates and distinct associations with HRQOL findings. Our data suggest that socio-economic status and comorbidities influence on their EWB. PMID- 19191022 TI - Estrogen receptor: is it predictive for response to cytotoxic as well as hormonal therapy? PMID- 19191023 TI - Expression of Gli1 correlates with the transition of breast cancer cells to estrogen-independent growth. AB - The failure of breast cancer treatment is largely due to the development of estrogen independence. Current data illustrate that Hedgehog (Hh) signaling may play an important role in breast cancer development. Here, we show that the expression of the Hh effector protein, Gli1 was significantly higher in estrogen independent breast cancer cells than in estrogen-dependent cells. Our data showed for the first time that stable expression of Gli1 in ER positive breast cancer cell lines MCF-7 and T47D can induce estrogen-independent proliferation and promote G1/S phase transition, which associated with cyclin-Rb axi. Gli1 can also attenuate the response of proliferation to estrogenic stimulation, which was correlated with down-regulation of expression of ERalpha and PR, as well as down regulation of transactivation of ERalpha. Our results suggest that up-regulation of Gli1 in breast cancer cells may be one of the mechanisms responsible for developing estrogen independence and this process may be regulated through down regulation of expression and transactivation of ERalpha. PMID- 19191024 TI - Lentiviral transfection of ependymal primary cultures facilitates the characterisation of kinocilia-specific promoters. AB - Ependymal primary cultures (EPCs) are an established model for studying ependymal cell biochemistry and the biology of kinocilia-bearing cells. However, the difficulty in causing them to express transgenes at high efficiency has been an important drawback of the system. Indeed plasmid-based transfection attempts remain at an efficiency below 1% and fail to elicit reporter gene expression, namely green fluorescent protein (GFP) synthesis, in any of the kinocilia-bearing cells of the cultures. Human immunodeficiency virus pseudotyped with the vesicular stomatitis virus envelope glycoprotein (HIV/VSV-G) and encoding GFP under the control of the ubiquitously recognised promoter of elongation factor 1 alpha (EF1alpha) also does not cause transgene expression in the kinocilia bearing cells of an EPC when applied at multiplicities of infection (MOIs) of up to 40 and destroys the culture when the MOI is increased further. In contrast, HIV/VSV-G encoding GFP under the control of a promoter specifically active in kinocilia-bearing cells leads to transgene expression in up to 79% of the kinociliated cells of an EPC when applied at an MOI of 20. This has permitted the initial characterisation of the promoter for the gene specifically transcribed in kinocilia-bearing cells, wdr16. The results have identified two regions of 100 nucleotides length each, which are critical for promoter activity and contain putative binding sites for the transcription factors Foxd1, Sox17 and Spz1. It appears that wdr16 is controlled by a bidirectional promoter also responsible for regulating the syntaxin 8 gene. PMID- 19191027 TI - "Unfortunately, we treat the chart:" sources of stigma in mental health settings. AB - BACKGROUND: Stigma within mental health settings may be equally detrimental to people with mental illnesses as societal stigma. AIMS: This study investigated stigma in mental health settings through a mixed qualitative-quantitative design. METHOD: Practitioners at a community mental health center indicated (1) their subjective experience of treating people with mental illness, and (2) descriptive features of people with mental illness. RESULTS: Interpretive phenomenological analysis found that a primary theme across practitioners was the causes and effects of labeling patients, a process practitioners attributed to other practitioners and/or to systemic pressures to "treat the chart" instead of the patient. Beyond symptoms and deficits, practitioners rated people with mental illnesses as "insightful" and "able to recover." CONCLUSIONS: These data suggest that stigma in mental health settings may be due to structural, systemic pressures on practitioners, with practitioners' emphasis on symptoms and deficits as a secondary factor. PMID- 19191025 TI - In vitro antioxidant activity of Valeriana officinalis against different neurotoxic agents. AB - Valeriana officinalis L. (Valerian) is widely used as a traditional medicine to improve the quality of sleep. Although V. officinalis have been well documented as promising pharmacological agent; the exact mechanisms by which this plant act is still unknown. Limited literature data have indicated that V. officinalis extracts can exhibit antioxidant properties against iron in hippocampal neurons in vitro. However, there is no data available about the possible antioxidant effect of V. officinalis against other pro-oxidants in brain. In the present study, the protective effect of V. officinalis on lipid peroxidation (LPO) induced by different pro-oxidant agents with neuropathological importance was examined. Ethanolic extract of valerian (0-60 microg/ml) was tested against quinolinic acid (QA); 3-nitropropionic acid; sodium nitroprusside; iron sulfate (FeSO4) and Fe2+/EDTA induced LPO in rat brain homogenates. The effect of V. officinalis in deoxyribose degradation and reactive oxygen species (ROS) production was also investigated. In brain homogenates, V. officinalis inhibited thiobarbituric acid reactive substances induced by all pro-oxidants tested in a concentration dependent manner. Similarly, V. officinalis caused a significant decrease on the LPO in cerebral cortex and in deoxyribose degradation. QA-induced ROS production in cortical slices was also significantly reduced by V. officinalis. Our results suggest that V. officinalis extract was effective in modulating LPO induced by different pro-oxidant agents. These data may imply that V. officinalis extract, functioning as antioxidant agent, can be beneficial for reducing insomnia complications linked to oxidative stress. PMID- 19191026 TI - Muscarinic inhibition of hippocampal and striatal adenylyl cyclase is mainly due to the M(4) receptor. AB - The five muscarinic acetylcholine receptors (M(1)-M(5)) are differentially expressed in the brain. M(2) and M(4) are coupled to inhibition of stimulated adenylyl cyclase, while M(1), M(3) and M(5) are mainly coupled to the phosphoinositide pathway. We studied the muscarinic receptor regulation of adenylyl cyclase activity in the rat hippocampus, compared to the striatum and amygdala. Basal and forskolin-stimulated adenylyl cyclase activity was higher in the striatum but the muscarinic inhibition was much lower. Highly selective muscarinic toxins MT1 and MT2-affinity order M(1) > or = M(4) >> others-and MT3 highly selective M(4) antagonist-did not show significant effects on basal or forskolin-stimulated cyclic AMP production but, like scopolamine, counteracted oxotremorine inhibition. Since MTs have negligible affinity for M(2), M(4) would be the main subtype responsible for muscarinic inhibition of forskolin-stimulated enzyme. Dopamine stimulated a small fraction of the enzyme (3.1% in striatum, 1.3% in the hippocampus). Since MT3 fully blocked muscarinic inhibition of dopamine-stimulated enzyme, M(4) receptor would be responsible for this regulation. PMID- 19191028 TI - Secondary resistance to cabergoline therapy in a macroprolactinoma: a case report and literature review. AB - Primary resistance to dopamine agonists occurs in 10-15% of prolactinomas but secondary resistance following initial biochemical and anti-proliferative response is very rare and has only been hitherto described in four previous cases, two with bromocriptine and two with cabergoline. We describe a case of a 57-year-old woman who presented with a large macroprolactinoma with suprasellar extension. She was initially treated with bromocriptine therapy with a resolution of symptoms, marked reduction in prolactin concentration and complete tumour shrinkage; a response which was subsequently maintained on cabergoline. After 8 years of dopamine agonist therapy, her prolactin concentration began to rise and there was symptomatic recurrence of her tumour despite escalating doses of cabergoline up to 6 mg weekly. Non-compliance was outruled by observed inpatient drug administration. The patient underwent surgical debulking followed by radiotherapy with good response. This case adds to the previous two cases of secondary resistance to cabergoline therapy in prolactinomas a marked initial response. While the mechanism of secondary resistance remains unknown and not possible to predict, close observation of prolactinoma patients on treatment is necessary. PMID- 19191029 TI - The World Health Organization (WHO) histologic classification of thymomas: a reanalysis. AB - The histologic classification of thymomas has remained for many years an area of conflict. In recent years, the World Health Organization (WHO) presented a proposal for the classification of these tumors, which has been widely adopted in many countries throughout the world. Two successive versions of this classification have been presented in recent years; however, controversy still exists regarding its accuracy and usefulness in clinical practice. We present a review of the current WHO classification of thymomas, with a discussion of some of the inconsistencies and limitations and the possible alternatives to the current approach for the histopathologic classification of these tumors. PMID- 19191030 TI - Novel biodegradable polyester poly(propylene succinate): synthesis and application in the preparation of solid dispersions and nanoparticles of a water soluble drug. AB - Poly(propylene succinate) (PPSu) polymers of average molecular weights from 2,800 to 13,100 g/mol were synthesized and characterized with regard to crystallinity, thermal properties, and cytocompatibility. Higher molecular weight samples exhibited lower degree of crystallinity and melted at lower temperatures. Melting of the polymer appeared to begin at 38 degrees C. PPSu cytocompatibility was investigated based on human umbilical vein endothelial cells viability in the presence of increasing concentrations of polymer, and it was found that PPSu exhibited comparable cytocompatibility with poly(DL-lactide). The feasibility of applying PPSu as a drug carrier was shown for the first time, as solid dispersions and nanoparticles of sodium fluvastatin based in PPSu were prepared. Drug release rates decreased with increasing the molecular weight of PPSu in both solid dispersions and nanoparticles. For dispersions prepared from PPSu of the same molecular weight, drug release rates increased with drug loading. It appears that PPSu applicability as a drug carrier warrants further consideration. PMID- 19191032 TI - Expression of Nkx2.5 in wild type, cardiac mutant, and thyroxine-induced metamorphosed hearts of the Mexican axolotl. AB - Nkx2.5, a homeodomain-containing transcription factor, is known to be necessary for normal heart development in vertebrates. It is one of the earliest lineage restricted genes expressed in cardiovascular progenitor cells and knowledge of its expression patterns has important therapeutic implications for damaged cardiomyocytes. Mexican axolotl is a unique system to study heart development for two reasons: the presence of a mutant phenotype lacking organized myofibrils due to sarcomeric tropomyosin deficiency and the ability to induce metamorphosis by administration of exogenous thyroid hormone. In this study, we cloned and sequenced the as yet uncharacterized Nkx2.5 cDNA from normal and cardiac mutant axolotl heart RNA. Comparison of cDNA sequences of Nkx2.5 from normal and mutant axolotl hearts did not show differences suggesting that loss of function mutation in Nkx2.5 is not responsible for the mutant phenotype. However, quantitative studies show higher expression of Nkx2.5 in mutant hearts raising the possibility that increased expression of Nkx2.5 may contribute to the mutant phenotype. We also evaluated quantitative changes in expression of Nkx2.5 in axolotl hearts during embryonic and postembryonic heart development induced by exogenous thyroid hormone. There is an apparent increase in Nkx2.5 transcript levels in metamorphosed hearts. PMID- 19191031 TI - Elastic liposomal formulation for sustained delivery of colchicine: in vitro characterization and in vivo evaluation of anti-gout activity. AB - Colchicine, an alkaloid found in extracts of the plants Colchicum autumnale and Gloriosa superb, is effective in the treatment of acute gout and dermatological conditions like leuko-cytoclastic vasculitis, psoriasis, and Sweet's syndrome. Oral administration of colchicine is associated with gastrointestinal side effects and its accumulation in the body leads to bone marrow suppression. In the present study, an attempt has been made for development and in vitro and in vivo evaluation of elastic liposomal formulation for topical delivery of colchicine. The in vitro skin permeation study across rat skin found transdermal flux for different elastic liposomal formulations to range between 32.8 +/- 1.2 and 44.4 +/- 1.9 microg h(-1) cm(-2), which was approximately seven to 11 times higher than obtained using drug solution (4.3 +/- 0.6 microg h(-1) cm(-2)). The results of skin deposition study showed that elastic liposomal formulation provide 12.5 fold higher skin deposition as compared to drug solution of colchicine. Confocal laser scanning microscopy also revealed better accumulation and deeper penetration (up to 200 microm) of elastic liposomes than drug solution (up to 12 microm). The biological evaluation of various vesicular formulations and drug solution was carried out using monosodium urate-induced air pouch model. The results of anti-gout activity in rats showed better and sustained biological effects in 24 h measured in terms of exudate volume (63.1 +/- 5.7% and 9.6 +/- 0.5% reduction with elastic liposomes and drug solution, respectively), reduction in leukocyte count (74.2 +/- 6.0% and 4.1 +/- 0.3% reduction with elastic liposomes and drug solution, respectively), decrease in inflammatory cells accumulation, and collagen deposition with elastic liposomal formulation than drug solution. Hence, the present study reveals that elastic liposomal formulation of colchicine possesses greater potential to enhance skin accumulation, prolong drug release, and improve the site-specificity of colchicine. PMID- 19191033 TI - Expression profile and distribution of Efhc1 gene transcript during rodent brain development. AB - One of the putative causative genes for juvenile myoclonic epilepsy (JME) is EFHC1. We report here the expression profile and distribution of Efhc1 messenger RNA (mRNA) during mouse and rat brain development. Real-time polymerase chain reaction revealed that there is no difference in the expression of Efhc1 mRNA between right and left hemispheres in both species. In addition, the highest levels of Efhc1 mRNA were found at intra-uterine stages in mouse and in adulthood in rat. In common, there was a progressive decrease in Efhc1 expression from 1 day-old neonates to 14-day-old animals in both species. In situ hybridization studies showed that rat and mouse Efhc1 mRNAs are expressed in ependymal cells of ventricle walls. Our findings suggest that Efhc1 expression is more important during initial phases of brain development and that at this stage it could be involved in key developmental mechanisms underlying JME. PMID- 19191034 TI - Lack of association between interleukin-1a gene (IL-1a) C (-889) T variant and polycystic ovary syndrome in Chinese women. AB - Polycystic ovary syndrome (PCOS) is associated with the spontaneous development of an aberrant heterogeneous hormonal environment that thought to be related to multiple genetic or environmental factors remain undefined. Previous studies indicated that IL-1a gene C (-889) T, a polymorphism of the gene encoding a protein which involved in various immune responses, inflammatory processes and hematopoiesis, and is associated with PCOS. A total of 205 PCOS patients and 177 healthy controls were included in this study that used PCR-RFLP to detect C ( 889) T variant of IL-1a gene. Clinical measures determined previously were included in the SPSS analysis. The results showed that the presence of IL-1a gene C (-889) T polymorphism in PCOS and healthy controls in Chinese women was not significantly different when studying genotype and allele frequencies. (Genotype: chi(2) = 4.62, df = 2, P = 0.10; allele: chi(2) = 1.35, df = 1, P = 0.25). Furthermore, no association was found between metabolic parameters observed and IL-1a genotypes in PCOS patients in this study. In conclusion, IL-1a gene C ( 889) T polymorphism does not appear to be risk factor for PCOS in this population of Chinese women. PMID- 19191035 TI - Aspirin prevents diabetic oxidative changes in rat lacrimal gland structure and function. AB - The aim of this study is to evaluate whether aspirin reduces Diabetis Mellitus (DM) oxidative damage in the lacrimal gland (LG), and ocular surface (OS). Ten weeks after streptozotocin induced DM and aspirin treatment, LG and OS of rats were compared for tear secretion, hidtology, peroxidase activity, and expression of uncoupling proteins (UCPs). DM reduction of tear secretion was prevented by aspirin (P < 0.01). Alterations of LG morphology and increased numbers of lipofucsin-like inclusions were observed in diabetic but not in aspirin-treated diabetic rats. Peroxidase activity levels were higher and UCP-2 was reduced in DM LG but not in aspirin treated (P = 0.0025 and P < 0.05, respectively). The findings prevented by aspirin indicate a direct inhibitory effect on oxidative pathways in LG and their inflammatory consequences, preserving the LG structure and function against hyperglycemia and/or insulin deficiency damage. PMID- 19191036 TI - Cushing's syndrome caused by an ACTH-producing ovarian steroid cell tumor, NOS, in a prepubertal girl. AB - Ectopic ACTH syndrome is a very rare cause of pediatric Cushing's syndrome. And if present, bronchial or thymic carcinoids predominate as causes. We hereby demonstrate a first case report of ACTH-producing ovarian steroid cell tumor, NOS, causing ectopic ACTH syndrome in a prepubertal girl. PMID- 19191037 TI - Changing the metal binding specificity of superoxide dismutase from Thermus thermophilus HB-27 by a single mutation. AB - Metal binding of superoxide dismutase from Thermus thermophilus HB27 was analyzed by comparing the related structures and sequences from different origins. Mutants (Ile166Leu, Asp167Glu, and Ile166Leu-Asp167Glu) were prepared and characterized. The mutants Asp167Glu and Ile166Leu-Asp167Glu changed their binding specificities from manganese to iron, which were manifested by the differences in color of the enzyme solutions and by flame atomic absorption analysis. Specific activities of the three mutants were 112, 52, and 62% of that of the wild-type enzyme, respectively. Asp167Glu and Ile166Leu-Asp167Glu only retained 6.8 and 6.1%, respectively, of the original activities after dialysis against 1 mM EDTA. Tryptophan fluorescence measurement and native gel electrophoresis implied that the three mutants could fold into a less condensed structure. Their folding and changes in the ion binding sites of the modeled structures might be the reason for their low affinities to metal ions. These findings increased our understanding of metal binding specificity of superoxide dismutase. PMID- 19191038 TI - Generation of Vibrio anguillarum ghost by coexpression of PhiX 174 lysis E gene and staphylococcal nuclease A gene. AB - Vibrio anguillarum ghosts (VAG) were generated, for the first time, using a conjugation vector containing a ghost bacteria inducing cassette, pRK-lambdaP(R) cI-Elysis, in which the expression of PhiX174 lysis gene E was controlled by the P ( R )/cI regulatory system of lambda phage. By scanning electron microscopy, holes ranging 80-200 nm in diameter were observed in the VAG. To avoid the presence of bacterial genomic DNA and an antibiotic resistance gene in the final VAG product, we constructed a new dual vector, pRK-lambdaP(R)-cI-E-SNA, containing the E-mediated lysis cassette and the staphylococcal nuclease A (SNA) mediated DNA degradation cassette, and generated safety-enhanced VAG for use as a fish vaccine. PMID- 19191041 TI - Gender differences in HIV and hepatitis C related vulnerabilities among aboriginal young people who use street drugs in two Canadian cities. AB - OBJECTIVES: Vulnerability to HIV and Hepatitis C virus (HCV) infection for indigenous populations worldwide must be contextualized in experiences of current and past trauma. Aboriginal women entrenched in poverty face further gender specific harms which place them at increased risk for HIV infection. METHODS: This study was cross-sectional and based on a community-based sample of Aboriginal young people (Metis, Aboriginal, First Nations, Inuit, and non-status Indians) between the ages of 14 and 30 years who used injection or non-injection non-cannabis illegal drugs (street drugs) in the previous month. Between October 2003 and July 2005, 543 participants living in either Vancouver or Prince George, Canada, were recruited by word of mouth, posters, and street outreach. Young people in the study completed a questionnaire administered by Aboriginal interviewers. Female participants (n = 262) were compared to male participants (n = 281) with respect to sociodemographics, trauma, sexual risk variables, and drug use patterns. Trained nurses drew blood samples for HIV and HCV antibodies and provided pre- and post-test counseling. RESULTS: Proportions positive for HIV and HCV were significantly higher among young women. HIV was 13.1% [9.5, 17.7] in women compared to 4.3% [2.5, 7.4] in men, and HCV was 43.6% [37.6, 49.8] in women as compared to 25.4% [20.5, 30.9] in men. When the analysis was restricted to young people who reported injection drug use, the proportions positive for HIV and HCV remained significantly higher among young women. Experiences of forced sex were reported by 70% of young women compared to 29% of young men, p < 0.001, while the median age of first forced sex was 6-years-old for both men and women. DISCUSSION: The results of the final model indicated that HIV had been associated with residing in Vancouver, having injected for longer, and sexual abuse, but not being female. However, this gendered analysis demonstrated that a greater proportion of young women were experiencing sexual abuse, and sexual abuse was associated with HIV positive status. Harm reduction and drug treatment programs are urgently required that target women at a young age and address complex traumatic experiences associated with childhood sexual abuse. PMID- 19191042 TI - Does time spent on household activities or housework stress complicate recovery following coronary artery bypass surgery? AB - OBJECTIVES: The purpose of the present study was to examine the role of household activities and housework stress as factors contributing to gender differences in recovery following coronary artery bypass graft (CABG) surgery. Therefore, the impact of time spent on household activities and housework stress on physical functioning (PF) was explored. METHODS: Psychosocial data were measured two months and again one year after CABG in a sample of 351 patients (23% women) recruited within one week before surgery. Self-report questionnaires included assessment of household responsibilities, depression (PHQ-9), and PF (SF-36 subscale). RESULTS: The results revealed that, following CABG, women spent more time on household activities and experienced markedly more stress due to these activities than men. Surprisingly, only housework stress was associated with poorer PF, whereas the time spent on household activities did not explain differences in PF in men and women. Finally, household responsibilities were not associated with the poorer recovery of the female CABG patient. DISCUSSION: In conclusion, our results do not support the advice that women should not resume household activities two months after surgery. More research is needed to establish valid guidelines. PMID- 19191046 TI - Retraction of authorship. PMID- 19191047 TI - Malignant arrhythmia: a case report of nasal NK/T-cell lymphoma with cardiac involvement. PMID- 19191039 TI - Botanical phenolics and brain health. AB - The high demand for molecular oxygen, the enrichment of polyunsaturated fatty acids in membrane phospholipids, and the relatively low abundance of antioxidant defense enzymes are factors rendering cells in the central nervous system (CNS) particularly vulnerable to oxidative stress. Excess production of reactive oxygen species (ROS) in the brain has been implicated as a common underlying factor for the etiology of a number of neurodegenerative diseases, including Alzheimer's disease (AD), Parkinson's disease (PD), and stroke. While ROS are generated by enzymatic and nonenzymatic reactions in the mitochondria and cytoplasm under normal conditions, excessive production under pathological conditions is associated with activation of Ca(2+)-dependent enzymes including proteases, phospholipases, nucleases, and alterations of signaling pathways which subsequently lead to mitochondrial dysfunction, release of inflammatory factors, and apoptosis. In recent years, there is considerable interest to investigate antioxidative and anti-inflammatory effects of phenolic compounds from different botanical sources. In this review, we describe oxidative mechanisms associated with AD, PD, and stroke, and evaluate neuroprotective effects of phenolic compounds, such as resveratrol from grape and red wine, curcumin from turmeric, apocynin from Picrorhiza kurroa, and epi-gallocatechin from green tea. The main goal is to provide a better understanding of the mode of action of these compounds and assess their use as therapeutics to ameliorate age-related neurodegenerative diseases. PMID- 19191048 TI - Reversible myocardial dysfunction in a young woman with metastatic renal cell carcinoma treated with sunitinib. PMID- 19191049 TI - TORP vs round window implant for hearing restoration of patients with extensive ossicular chain defect. AB - CONCLUSIONS: Round window implant (RWI) with a floating mass transducer (FMT) may be suggested as the first choice in hearing rehabilitation for patients with chronic otitis media (COM) and extensive destruction of the ossicular chain. OBJECTIVE: To investigate the pros and cons of the total ossicular replacement prosthesis (TORP) vs the RWI in restoration of hearing in patients with COM. PATIENTS AND METHODS: Thirty-eight patients with bilateral moderate to severe mixed or conductive hearing loss from COM without cholesteatoma and bilateral ossicular chain erosion (footplate residual) were alternately assigned to a titanium-TORP (T-TORP) and to RWI with the FMT of the Medel Vibrant Soundbridge (MVBS) located onto the RW niche. The therapeutic efficiency, preoperative vs postoperative air-conduction gain and speech recognition were investigated for the two groups and statistically analyzed at 36 months postoperatively. The following postoperative anatomic conditions were also evaluated for the two groups: 1) recurrence of infection, 2) retraction pocket, 3) extrusion rate, and 4) displacement of the prosthesis. RESULTS: Good functional results and stability at 36 months were obtained with both procedures. The extrusion rates for T-TORP were low. So far no extrusion has been observed for RWI. Hearing results were statistically much better for RWI vs T-TORP for all investigated parameters. PMID- 19191051 TI - Imaging of calvarial lesions. PMID- 19191052 TI - Clinical review: impact of statin substitution policies on patient outcomes. AB - BACKGROUND: The increasing awareness of cost issues in health care has led to the increasing use of policy-driven substitution of branded for generic medications, particularly relative to statin treatment for cardiovascular diseases. While there are potential short-term health care savings, the consequences for primary care are under-researched. Our objective was to review data on intensive statin therapy and generic substitution in patients at high cardiovascular risk. RESULTS: Current treatment guidelines for the prevention of cardiovascular disease are consistent in their recommendations regarding statin therapy and treatment targets. Clinical trials demonstrate that to reduce cardiovascular events, a statin is more effective than placebo, intensive statin therapy is more effective than moderate statin therapy in patients with established coronary disease, and in patients receiving intensive statin therapy the lowest risk is associated with the lowest low-density lipoprotein levels. However, in clinical practice, patients at high cardiovascular risk are prone to be undertreated. Observational studies suggest that mandatory statin substitution may increase the gap between achieved and recommended therapeutic targets. CONCLUSIONS: Substitution of generic statins may be cost-saving, particularly at the primary prevention level. However, statin substitution policies have not been adequately studied on a population level. Data raise concern that mandated statin substitution may lead to unfavourable treatment choices at the level of the individual high-risk patient. PMID- 19191053 TI - Metabolomics: unraveling the chemical individuality of common human diseases. AB - Sir Archibald Garrod is often referred to in recent perspectives on metabolomics because he was the first to recognize 'inborn errors of metabolism'. For decades, the determination of metabolites was the domain of those involved in the diagnosis of this class of inherited disorders. With the development of metabolomics, these methods to determine and analyze metabolites have been taken an exciting step forward and are now used to understand common human disease. This concept of looking at metabolites to solve the pathogenesis of human disease touches upon another concept developed by Garrod, known as 'chemical individuality'. Garrod proposed that each person is biochemically unique due to inherited differences in enzymes, which is reflected in disease predisposition. In a more contemporary perspective, this concept may be extended to chemical individuality of a human disease. This is the domain of metabolomics, which aims to determine as many metabolites as possible in samples from a cohort of individuals. Analysis of the results will identify changes in metabolites that correlate with the presence of certain afflictions. The next challenging step is then to determine whether these metabolites are only biomarkers for the presence of a disease or new leads to an unknown etiology. PMID- 19191054 TI - A new induction protocol for the control of steroid refractory/dependent acute graft versus host disease with alefacept and tacrolimus. AB - BACKGROUND AIMS: We have shown previously that alefacept is effective in acute steroid resistant/dependent and chronic extensive graft versus host disease (GvHD) with a protocol using timings similar to those used for psoriasis treatment. In this study, we describe the use of an alefacept induction (e.g. for 7 consecutive days) followed by a bi-weekly maintenance treatment in combination with tacrolimus for acute steroid resistant/dependent GvHD 1, 3. METHODS: Sixteen patients were treated in this cohort, most with refractory GvHD. The pre treatment GvHD grade ranged from 2 to 4 (median 3), involving the skin 16, gut 11 and liver 5. RESULTS: Twelve out of the 16 patients showed a response. As with the first protocol, the response of GvHD in the skin was fastest. In contrast to our previous protocol, however, the gastro-intestinal (GI) GvHD response was faster (P=0.05 compared with the first cohort). A hepatic response was seen in 4/6 patients and was complete in three. All responses were durable, including mucocutaneous, gut and liver GvHD. In all responding patients we were able to decrease the steroid dose significantly and in seven it was completely withdrawn. CONCLUSION: Alefacept induction is safe in acute steroid resistant/dependent GvHD and may be more effective therapeutically than our previous alefacept protocol. We speculate that alefacept initiates an allo-versus-allo cellular effect through its Fc receptor. PMID- 19191055 TI - Cell therapy for disorders of bone. AB - Bone marrow transplantation (BMT) has changed the course of treatment for an array of diseases, including disorders of bone. Hematopoietic stem cells (HSC) within the marrow are known to be the precursors of osteoclastic bone cells, and trials of BMT in osteopetrosis, a disorder characterized by a deficiency of osteoclasts, have resulted in significant clinical improvement in patients. The origin of the other major bone cell, the osteoblast, remains uncertain, although studies have identified osteoprogenitor cells within the marrow, leading to further investigation of both mesenchymal stromal cells (MSC) and HSC as candidates for this role. A better understanding of the source of osteoblasts and normal bone metabolism is crucial to efforts to develop effective cell therapy for bone disorders characterized by deficient or abnormal osteoblast function. This review focuses on systemic and local cell therapy in the treatment of several genetic bone disorders and osteoporosis, an acquired disorder caused by abnormal bone metabolism, with the intent of presenting both the progress and challenges associated with this emerging form of therapy. Although the risks of systemic transplantation must be carefully considered, cell therapy for disorders of bone carries the potential for long-term and potentially curative benefits, justifying further intensive research on this important treatment option. PMID- 19191056 TI - Labeling of human mesenchymal stromal cells with superparamagnetic iron oxide leads to a decrease in migration capacity and colony formation ability. AB - BACKGROUND AIMS: Labeling of stem cells is crucial to allow tracking of stem cell homing and engraftment after transplantation. In this study we evaluated the influence of cell labeling procedures using clinically approved small particles of iron oxide (SPIO) with or without transfection reagents (TA) on functional parameters of human mesenchymal stem cells (MSC). METHODS: The study was approved by the institutional review board of the University of Tubingen, Germany. Seven populations of bone marrow (BM)-derived human mesenchymal stem cells (MSC) were labeled with SPIO alone or in combination with various TA. Directly after labeling and two passages after labeling migration assays, quantification of colony-forming units and quantitative evaluation of the differentiation potential were performed. Quantification of the cellular total iron load (TIL), determination of the cellular viability and electron microscopy were also performed. RESULTS: Labeling of mesenchymal stem cells with SPIO with or without TA did not affect cell viability and differentiation potential significantly. SPIO in combination with TA coated the cellular surface directly after labeling but was incorporated into the cells after two passages. Labeling of mesenchymal stem cells with TA led to a significant decrease of migration capacity. This effect was abolished after two passages. Labeling with and without TA led to a significant decrease in colony formation ability. This effect could also be observed after two passages. CONCLUSIONS: The observed decrease of migration capacity and colony-formation ability was not associated with either TIL or localization of particles of iron oxide. SPIO labeling with and without TA had functional effects on human mesenchymal stem cells by decreasing the migration capacity and colony-formation ability of the stem cells. PMID- 19191057 TI - Reinnervation cannot be interpreted as an indicator of electrophysiologic improvement in amyothrophic lateral sclerosis. AB - Denervation and reinnervation comprise a dynamic process that begins in the early periods of amyothrophic lateral sclerosis and lasts until the final individual motor neuron dies. A dynamic process like denervation-reinnervation that is found in the natural course of the disease can not be presented as a neurophysiologic measure. Other parameters, such as motor unit number estimation, compound muscle action potential amplitude and neurophysiologic index, seem more appropriate for therapeutic trials. PMID- 19191058 TI - Stem-cell transplantation into the frontal motor cortex in amyotrophic lateral sclerosis patients. AB - BACKGROUND AIMS: Amyotrophic lateral sclerosis (ALS) is characterized by the selective death of motor neurons. CD133(+) stem cells are known to have the capacity to differentiate into neural lineages. Stem cells may provide an alternative treatment for ALS and other neurodegenerative diseases. METHODS: Five men and five women (aged 38-62 years) with confirmed ALS were included in this study. Our institutional ethics and research committees approved the protocol. After informed consent was obtained, patients underwent Hidrogen-Magnetic Resonance Imaging (H-MRI) spectroscopy and were given scores according to an ALS functional rating scale, Medical Research Council power muscle scale and daily living activities. Bone marrow was stimulated with 300 microg filgrastim subcutaneously daily for 3 days. Peripheral blood mononuclear cells were obtained after admission by leukapheresis. The cell suspension was conjugated with anti human CD133 superparamagnetic microbeads, and linked cells were isolated in a magnetic field. The isolated cells (2.5-7.5x10(5)) were resuspended in 300 microL of the patient's cerebrospinal fluid, and implanted in motor cortexes using a Hamilton syringe. Ten patients with confirmed ALS without transplantation were used as a control group. Patients were followed up for a period of 1 year. RESULTS: The autologous transplantation of CD133(+) stem cells into the frontal motor cortex is a safe and well-tolerated procedure in ALS patients. The survival of treated patients was statistically higher (P=0.01) than untreated control patients. CONCLUSIONS: Stem-cell transplantation in the motor cortex delays ALS progression and improves quality of life. PMID- 19191059 TI - Assessment of quality of life and participation within an outpatient rehabilitation setting. AB - PURPOSE: To evaluate the use of quality of life and participation measures in routine rehabilitation practice, determine the relationship between two scales and investigate their ability to evaluate change in a general outpatient population over a time of rehabilitation. METHODS: This cohort study consisted of 55 patients admitted consecutively to an outpatient rehabilitation centre. All participants completed the WHOQOL-BREF and the London Handicap Scale (LHS) at admission and discharge. RESULTS: Non-parametric statistical analysis demonstrated a significant change in the total scores of both the WHOQOL-BREF and LHS over time (Z = -4.33, P < 0.001 and Z = -4.40, P < 0.001 respectively) with 'better' scores on completion of rehabilitation. A strong correlation existed between the total WHOQOL-BREF and LHS scores both cross-sectionally (admission and discharge p > 0.7), and longitudinally (p = 0.63). CONCLUSIONS: Our study demonstrated a strong relationship between quality of life and participation, with change shown using both scales over a period of outpatient rehabilitation. Given the findings, it seems appropriate to use only one of these measures. The WHOQOL-BREF has a wider scope, being useful both to evaluate service delivery in a diverse outpatient population or when measuring an individual over time. PMID- 19191060 TI - World Health Organisation Disability Assessment Schedule II: contribution to the Italian validation. AB - INTRODUCTION: The World Health Organisation Disability Assessment Schedule II (WHODAS II) is an instrument developed by the World Health Organisation in order to assess behavioural limitations and restrictions to participation experienced by an individual, independently from a medical diagnosis. The conceptual frame of reference of this instrument is the International Classification of Functioning, Disability and Health: ICF. Specifically, the instrument is designed to evaluate the functioning of the individual in six activity domains: Understanding and communicating, Getting around, Self-care, Getting along with people, Life activities, Participation in society. PURPOSES: Considering the widespread consent about the usefulness of the WHODAS II, the general aim of the present study is to provide a contribution to the validation of the Italian version. METHOD: The WHODAS II Italian version has been administered to a sample of 500 participants subdivided in two groups: 271 normal adults and 229 disabled adults. The disabled participants group has been subdivided in three sub-groups, according to their disability: 111 motor disabled, 45 mental disabled, 73 sensory disabled. RESULTS: The mean Total score of the WHODAS II is 12.95 for the normal adults and 22.93 for the disabled group. Either group obtains the least impairment in the Self-care domain. This could be probably due to the presence of social-health workers in everyday life for all Italian disabled people. For the three disabled participant groups separately computed the mean Total score is: 28.66 for the motor disabled, 24.60 for the mental disabled, and 14.97 for the sensory disabled, confirming that sensory disabled do not perceive their disability as a personal functioning problem but a socially constructed one. Some subscales of WHODAS II show relatively strong floor effects. The Cronbach's Alpha calculated for each of the subscales is found to be high. The correlations of the subscales show strong correlations in all subscales. CONCLUSIONS: The WHODAS II is a useful instrument for measuring disability and functioning in normal and disabled people. It shows high reliability and a stable factor structure; although an additional psychometric evaluation of a representative sample of Italian disabled should be carried out in order to reach standard scores for each macro-category of disability. PMID- 19191061 TI - Exploratory findings with virtual reality for phantom limb pain; from stump motion to agency and analgesia. AB - PURPOSE: Phantom limb pain is chronic and intractable. Recently, virtual reality (VR) and motion capture technology has replicated the mirror box device of Ramachandran (Ramachandran et al. Nature 1995, 377, 489-490; Ramachandran and Rogers-Ramachandran Proc R Soc Biol Sci 1996, 263, 377-386) and led to reductions in this pain. We present results from a novel variation on this method which captures motion data directly from a patient's stump (rather than using the opposite remaining limb) and then transforms it into goal directed, virtual action enacted by an avatar in a VR environment. METHOD: A sample of subjects with 'arm' (n = 7) and 'leg' (n = 7) amputations underwent trials of a virtual reality (VR) system, controlled by motion captured from their stump which was translated into movements of a virtual limb within the VR environment. Measures of pain in the phantom limb were elicited from patients before and during this exercise as they attempted to gain agency for the movement they saw, and feel embodied within the limb. After this each subject was interviewed about their experiences. RESULTS: Five subjects in each group felt the virtual limb to be moved by them and felt sensations of movement within it. With this they also reported reductions in their phantom limb pain greater than expected from distraction alone. No carry over effect was seen. CONCLUSIONS: This technique, which has shown similar success rates to trials of a virtual mirror box, is relatively cheap and portable, and will allow further trials in a home environment. PMID- 19191062 TI - Wheelchair accessibility to public buildings in Istanbul. AB - BACKGROUND: Accessibility to public environment is the human right and basic need of each citizen and is one of the fundamental considerations for urban planning. PURPOSE: The aim of this study is to determine the compliance of public buildings in central business districts (CBD) of Istanbul, Turkey, to wheelchair accessibility to the guidelines of the instrument and identify architectural barriers faced by wheelchair users. METHODS: This is a descriptive study of 26 public buildings in CBD of Istanbul. The instrument used is the adapted Useh, Moyo and Munyonga questionnaire to collect the data from direct observation and measurement. Descriptive statistics of simple percentages and means are used to explain the compliance to the guidelines of the instrument and wheelchair accessibility. RESULTS: The descriptive survey results indicate that wheelchair users experience many accessibility problems in public environment of the most urbanised city (cultural capital of Europe in 2010) in a developing country. CONCLUSION: It is found that the major architectural barrier is the public transportation items with the lowest mean compliance (25%). Beside this, the most compliant to the instrument is entrance to building items with 79% as mean percentage. It is also found that there is an intention to improve accessibility when building construction period is investigated. This article describes the example of the compliance of public buildings accessibility when the country has legislation, but lacking regulations about accessibility for the wheelchair users. PMID- 19191063 TI - Criterion-related validity of the one-mile run/walk test in children aged 8-17 years. AB - We assessed the criterion-related validity of Cureton's equation for estimating peak oxygen consumption (VO(2peak)) from the one-mile run/walk test in endurance trained children aged 8-17 years. Altogether, 66 physically active white children and adolescents (32 girls, 34 boys) completed a graded exercise test to volitional exhaustion and the one-mile run/walk test. Cureton's equation was used to estimate VO(2peak), and was assessed using several error measures. Agreement between measured VO(2peak) and estimated VO(2peak) was analysed by the Bland and Altman method. The correlation coefficient between measured VO(2peak) and one mile run/walk time was -0.59 (P < 0.001) and that between measured and estimated VO(2peak) was 0.70 (P < 0.001). The mean difference between measured and estimated VO(2peak) was 10 ml . kg(-1) . min(-1) (95% CI = 9.2-11.8; P < 0.001). The standard error of the estimate was 3 ml . kg(-1) . min(-1), and the percentage error was 32%. There was a positive association between the measured and estimated VO(2peak) difference and the measured and estimated VO(2peak) mean, which indicates that the higher the VO(2peak) the higher the error of the estimate. These findings did not change markedly when the analyses were performed by sex, age group or body mass status. These results suggest that Cureton's equation systematically underestimates VO(2peak) in endurance-trained children with high VO(2peak). PMID- 19191064 TI - Understanding parental stressors: an investigation of British tennis-parents. AB - In this study, we examined the stressors experienced by British tennis-parents. The parents (n = 123) completed an extensive survey focused on the internal and external demands that they had encountered through having a child compete in the sport. The survey consisted of open-ended questions related to competition, coaching, organizational, personal, and developmental issues. Inductive and deductive content analysis resulted in the development of seven core themes of tennis-parental stressor: competition, coaches, finance, time, siblings, organization-related, and developmental. Parents experienced a diverse number of competitive stressors indicating the particular difficulties they faced before, during, and after matches involving their child, opponents, other parents, and officials. They also reported a wide range of organizational stressors that paralleled the financial, social, and personal investments that accompanied their support roles. The results of this research reinforce the importance of parents possessing the necessary skills to cope with the psychological, developmental, and logistical demands of competitive tennis. Implications with respect to induction workshops and education for coaches and parents are presented, as well as consideration for governing bodies to enhance their communication channels and logistical support. Future research recommendations are posed to build upon the study of this domain in youth sport. PMID- 19191065 TI - Imagery and observational learning use and their relationship to sport confidence. AB - The present study investigated 345 athletes' (male = 152, female = 193) use of observational learning and imagery for practice and at competition and how this related to sport confidence. The Functions of Observational Learning Questionnaire (Cumming et al., 2005), the Sport Imagery Questionnaire (Hall et al., 1998), and the Trait Sport Confidence Inventory (Vealey, 1986) were contextualized by asking participants to rate each item twice, once for practice and once for competition. The athletes reported using each of the different functions of observational learning and imagery in these situations, but the pattern of use differed. Whereas nearly all of the imagery functions were more frequently used at competition, the majority of observational learning functions were used more for practice. Cognitive specific and motivational general-mastery imagery were significant predictors of sport confidence in practice and competition, whereas the skill function of observational learning significantly predicted practice confidence only. PMID- 19191066 TI - Improved single-letter identification after whole-word training in pure alexia. AB - Pure alexia is characterised by a very time-consuming letter-by-letter reading strategy due to an impaired identification or integration of single letters. So far, therapy interventions have addressed impaired letter identification using specific single-letter training approaches. In the present study, we report patient KA with pure alexia and letter-by-letter reading. Contrary to common approaches, we investigated whether a whole-word reading approach can be successful despite severe impairments at the level of single-letter identification. As a first step, auditory-visual verification tasks were used to familiarise the patient with the training items, which in a second step were read with limited exposure duration. After a four week intervention, KA's word reading performance improved significantly for trained and control items in terms of speed and accuracy. Although not specifically addressed during our training programme, even single-letter identification and text reading performance increased significantly. However, the patient's reading was still based on a letter-by-letter strategy. PMID- 19191067 TI - Reassessment of a classical single injection 51Cr-EDTA clearance method for determination of renal function in children and adults. Part II: Empirically determined relationships between total and one-pool clearance. AB - BACKGROUND: The one-pool or slope-intercept technique is widely used when determining total (51)Cr-EDTA plasma clearance (Cl). The one-pool clearance (Cl(1)), which always exceeds Cl, has mostly been corrected to Cl by multiplication by a constant factor = 0.80, suggested by Chantler (CH(0.80)), or by using a second-order polynomial originally proposed by Brochner-Mortensen (BM) and later recommended by the British Nuclear Medicine Society (BM(BNMS)). Theoretical considerations indicate that the CH correction gives a systematic overestimate of Cl, whereas the BM correction may underestimate Cl at high values. OBJECTIVE: To assess the accuracy of Cl as estimated from Cl (1) corrected either by CH(0.80) or by second-order polynomials. MATERIAL AND METHODS: Cl(ref) was determined in 149 subjects (M/F/children: 71/46/32) from a complete plasma curve followed for 4-5 h after injection of (51)Cr-EDTA (range of Cl(ref) : 8-183 mL/min/1.73 m(2)). Cl(est) was determined from Cl(1) subsequently corrected by CH(0.80) and four second-order polynomials. RESULTS: Using CH(0.80) correction, Cl(est) underestimated Cl(ref) (by a maximum of 20%) at Cl(ref) values less than about 100 mL/min/1.73 m(2) in children and 130 mL/min/1.73 m(2) in adults. At higher clearance levels, Cl(ref) was increasingly overestimated. Taking the BM(BNMS) correction as representative of second-order polynomials, Cl(est) increasingly underestimated Cl(ref) at high levels, the error being 10% at a Cl(ref) value of about 175 mL/min/1.73 m(2). CONCLUSIONS: We suggest that the tested correction equations are replaced by the given common correction equation based on the "true" relationship between Cl(1) and Cl thoroughly described in part I of this study. PMID- 19191068 TI - Plasma predilution with addition of depleted plasma in a prothrombin time reagent improves the agreement between different prothrombin time methods. AB - BACKGROUND: The Quick (plain thromboplastins) and Owren (combined thromboplastins, with the addition of bovine plasma with vitamin K-dependent factors depleted) prothrombin time (PT) methods are used for measuring prothrombin time; for instance, in monitoring anticoagulation therapy with vitamin K antagonists. In most Quick PT methods, the final plasma dilution is 1/3 compared to Owren methods, which have a final dilution of 1/21. The Quick PT methods are associated with larger inter-laboratory variations than the Owren PT methods. OBJECTIVES: We investigated whether dilution of the sample or thromboplastin has any impact on the correlation between the different PT methods. MATERIAL AND METHODS: Plasma samples were analysed undiluted and prediluted in buffer, with adsorbed bovine plasma added, and by using rabbit brain or human placenta thromboplastins. Bootstrapping (re-sampling) was utilized to estimate the difference in correlations between PT of different reagents at different dilutions of plasma. RESULTS AND CONCLUSIONS: We found the best correlations for combined reagents when the sample was prediluted 7-fold (r = 0.95) or more, and lowest when the sample was undiluted (r = 0.67). The source of thromboplastin had only a minor impact, if any, on the PT results. It was impossible to test whether predilution of plasma samples improved the correlation between Quick PT methods due to absence of clotting in many samples. We suggest that the Owren-style sample predilution is preferable for the harmonization of PT results and to overcome the large inter-laboratory variations that are associated with Quick PT methods to the benefit of patients on oral anticoagulation. PMID- 19191069 TI - Increased intragastric pressure gradients are involved in the occurrence of acid reflux in gastroesophageal reflux disease. AB - OBJECTIVE: Increased pressure gradients across the esophagogastric junction (DeltaEGJp) play a role in gastroesophageal flow during TLESR. The aim of this study was to further explore DeltaEGJp in patients with gastroesophageal reflux disease (GERD) and controls. MATERIAL AND METHODS: Twenty GERD patients were studied along with 20 control subjects. High resolution manometry and pH recording were performed 1 h before and 2 h after a liquid meal (500 ml/300 kcal). DeltaEGJp was calculated at the start of a TLESR and at 180, 60, and 10 s before TLESR. RESULTS: DeltaEGJp at the start of a TLESR and at 180, 60, and 10 s before TLESR was markedly increased in GERD patients compared with that in control subjects (9.9 mmHg and 7.5 mmHg, respectively; p<0.05). Whilst intragastric pressure gradients in GERD patients were increased compared with those in controls (4.6 mmHg and 2.5 mmHg, respectively; p<0.01), intraesophageal pressure gradients were similar in both groups. Furthermore, in controls, first- and second-hour postprandial intragastric pressures were decreased compared with in fasting periods (1.9 +/- 0.4 mmHg and 2.1 +/- 0.4 mmHg versus 3.5 +/- 0.4 mmHg; p<0.05), while this was not observed in GERD patients. CONCLUSIONS: In GERD patients, DeltaEGJp is greater than that in controls both before and during TLESR. This phenomenon is caused by increased intragastric pressure and might contribute to increased rates of acid reflux during TLESR in GERD patients. PMID- 19191070 TI - Emphysematous gastritis: a deadly infectious disease. AB - We report a case of emphysematous gastritis with secondary peritonitis. Despite aggressive surgical intervention and appropriate antimicrobial treatment, the patient ran a fulminant course leading to death. Review of the literature in the past 20 y provided additional information on this disease entity. PMID- 19191071 TI - The value and meaning of an instrumental occupation performed in a clinical setting. AB - The aim was to investigate how patients in a clinical setting, combining acute stroke care and rehabilitation, perceived the value and meaning attached to a commonly used instrumental occupation, namely "Brewing coffee and making an open face cheese sandwich with sliced vegetable". Another aim was to validate the ValMO model, proposing that value and meaning are related phenomena. Upon performing the occupation, 38 participants answered questionnaires concerning perceived meaning and occupational value. The results showed that the occupation was highly valued by the participants and that it was perceived as meaningful. No age or gender differences were found. The findings confirmed the proposed link between value and meaning. In conclusion, a commonly used and supposedly meaningful occupation was indeed found to be valued and meaningful by the patients and the findings validated the targeted aspects of the ValMO model. This study was unique in its focus on value and meaning in a clinical context. Futures studies should clarify if the link between meaningful occupation and well-being, which has been shown in many studies of everyday situations and is another of the assumptions in the ValMO model, can be proved in clinical contexts as well. PMID- 19191072 TI - Development, psychometric properties, and validity of the hopkins adult reading test (HART). AB - We describe the development of a 35-item, oral word-reading test with two equivalent forms (HART-A and HART-B) designed to estimate premorbid abilities. Both forms show excellent internal consistency (coefficients alpha>.91) and test retest reliability (Pearson rs >.90). HART performance was combined with demographic variables to generate regression equations that predict IQ scores obtained concurrently and 4-8 years earlier. The resulting models explained 61% of full scale IQ (FSIQ) variability in 327 healthy adults. The FSIQs that can be estimated range from below 73 to above 131. Combined with demographic variables, these two brief word reading tests accurately predict a broader range of IQs than Blair and Spreen's (1989) longer version. Equivalent forms make it especially useful for longitudinal studies. PMID- 19191073 TI - Gray's BIS/BAS dimensions in non-comorbid, non-medicated social anxiety disorder. AB - Gray's behavioural inhibition and behavioural activation (BIS/BAS) neural systems model has led to research on approach and withdrawal as the two most fundamental dimensions of affective behaviour, and their role in psychopathology. Although Gray proposed the BIS as the neurological basis of anxiety, there are no reports examining approach and withdrawal predispositions in social anxiety disorder. Here we report approach and withdrawal predispositions in a group of 23 non medicated individuals with social anxiety disorder (SAD) without co-morbid depression and in 48 normal controls. Results show increased BIS and decreased BAS fun-seeking in SAD subjects thereby underscoring Gray's dimensional model. PMID- 19191074 TI - Paternal age and common mental disorders. AB - INTRODUCTION: There is evidence in the literature that there are associations between advancing paternal age and psychosis or more specifically schizophrenia, but not enough to support a strong link between advancing paternal age and common mental disorders. OBJECTIVE: This study aims to explain the association between paternal age at birth and common mental disorders in progeny during their adulthood. METHODOLOGY: This is a sub-study from a larger survey which was planned to study the epidemiology of mental disorders in Malaysia. Respondents who could remember the age of parents at birth were included in the study. The diagnosis of common mental disorders (CMD) was made using the CIS-R (Clinical Interview Schedule-Revised) instrument in the PROQSY (Programmed Questionnaire System) format. Association between paternal age at birth and CMD was studied using logistic regression, after controlling for age, gender, ethnicity and presence of family history of mental disorders. RESULTS: Respondents with paternal age at birth of 19 and below and 50 above and had higher rates of 10 and 25% for common mental disorders (chi(2)=7.007, P=0.072) with odds ratios of 2.89 (95% CI of OR = 1.1-7.6) and 4.28 (1.4-12.7). DISCUSSION: Progenies of fathers under 20 and over 50 had higher risk for mental disorders. Factors such as immaturity in sperm of teenage fathers, mutation in germ line of older fathers, environmental and psychosocial factors could have contributed to increased prevalence of common mental disorders in the progeny. PMID- 19191075 TI - Improvement of survival in sex cord stromal tumors - an observational study with more than 25 years follow-up. AB - OBJECTIVE: To highlight aspects of malignant ovarian sex cord stromal tumors, effects of treatment, and developments over the past 28 years. DESIGN: Population based cohort study. SETTING: Gynecological departments within the catchment-area of the Munich Cancer Registry and associated with the project group 'Malignant Ovarian Tumors' of the Munich Cancer Center. SAMPLE: One hundred and forty-five women with an invasive single sex cord stromal tumor diagnosed between 1978 and 2005. METHODS: Overall survival was estimated with the Kaplan-Meier method, relative survival was computed by the ratio of observed to expected survival rate. The impact of age, International Federation of Gynecology and Obstetrics (FIGO)-stage, residual tumor, and chemotherapy was examined by multivariate analysis (Cox regression model). MAIN OUTCOME MEASURES: Overall and relative survival and multivariate adjusted overall survival. RESULTS: Survival data showed a five-/10-year overall survival of 55.8%/42.8% (relative survival 58.6%/49.2%) for women diagnosed before 1988 and 89.1%/78.3% (relative survival 92.7%/85.2%) for women diagnosed after 1988. After adjustment for age and FIGO stage, the following hazard ratios and 95% confidence intervals (95% CI) for treatment methods resulted: 3.3 (95% CI 1.5-7.0) for women with compared to women without residual tumor and 2.2 (95% CI 1.2-4.2) for women with chemotherapy compared to women where no chemotherapy was given. CONCLUSIONS: Improvements in survival may be attributed to a stage-shift toward more favorable stages at diagnosis and to advances in treatment such as improved surgery without residual tumor. There is no evidence for any benefit of adjuvant chemotherapy. Surgery remains the cornerstone of treatment, yet the benefit of postoperative therapy is still under debate. PMID- 19191076 TI - Perinatal risk factors in young adult-onset type 1 and type 2 diabetes - a population-based case-control study. AB - OBJECTIVE: To examine the effects of the size of the mother and the newborn, including placental weight and gestational age at delivery, on the risk for young adult-onset type 1 diabetes (T1DM) and type 2 diabetes (T2DM). DESIGN: Case control study. SETTING: Finland. POPULATION: Subjects with T1DM and T2DM aged 15 39 at diagnosis between the years 1992 and 1996. The number of case-control pairs was 858 for T1DM and 355 for T2DM. METHODS: Diabetic subjects were identified from the Finnish national healthcare registers and reports from diabetes nurses. Control subjects were obtained from the population register. Data on perinatal factors were obtained from the original healthcare records. The odds ratios (ORs) for both types of diabetes were estimated using conditional logistic regression. RESULTS: The risk for early-onset T2DM decreased with increasing birthweight until 4.2 kg (OR 0.49 (95% confidence interval 0.37-0.66) per 1 kg), but with birthweight above 4.2 kg the risk increased (OR 4.8 (1.3-17.6) per 1 kg). The risk for T2DM decreased also with increasing birth length (OR 0.88 (0.81-0.95) per 1 cm), body mass index at birth (OR 0.81 (0.73-0.90) per 1 kg/m2), and placental weight (OR 0.77 (0.61-0.98) per 100 g). The latter was not significant when adjusted for birthweight. The examined perinatal factors did not affect the risk for T1DM in young adults. CONCLUSIONS: Birth size significantly affects the risk for T2DM diagnosed in young adulthood but no evidence was found of an association between late-onset T1DM and perinatal factors. PMID- 19191077 TI - Maternal eating disorders and offspring sex ratios at birth. PMID- 19191079 TI - Large Meckel's diverticulum complicating pregnancy with an intestinal obstruction. PMID- 19191080 TI - Apolipoprotein AI and Apolipoprotein B gene polymorphisms and lipid profile in Tamilian population. AB - Apolipoprotein (ApoAI) is the major protein constituent of high density lipoproteins (HDL). Apolipoprotein (apo) B-100, a component of low density lipoprotein (LDL), serves as a ligand for the removal of LDL from the circulation by the LDL receptor. Genotyping of ApoAI and ApoB polymorphisms was carried out in 185 healthy Tamilian volunteers of south India after clinical examination. Lipid profile was estimated and polymorphisms were detected by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The frequency of the rare M1-, M2- and R- alleles were 21%, 4.3% and 5.4%, respectively. An increase of 9.1 mg dL(-1) (0.23 mmol L(-1)) in HDL-cholesterol (HDL-C) levels was observed with M1-/- genotype when compared to M1+/+ genotype, which was not found after adjustments were made for confounding risk factors. A paradoxical increase in levels of total cholesterol and LDL-cholesterol (LDL-C) was observed with M2+/- genotype when compared to M2+/+ genotype. Analysis of combination of genotypes of ApoAI revealed no influence on the lipid parameters. ApoB EcoRI in contrast to ApoAI polymorphisms had no significant effect on lipid profile.. PMID- 19191081 TI - Determinants of overweight associated with stunting in preschool children of Yaounde, Cameroon. AB - BACKGROUND: Nutrition transition in developing countries has been associated with higher prevalence of overweight. AIM: The study aimed to identify the factors associated with concurrent stunting and overweight in urban preschool children of Yaounde, Cameroon. SUBJECTS AND METHODS: 169 preschool children of both sexes were recruited according to their nutritional status: Stunted, overweight, stunted-overweight, and non-stunted-non-overweight. Factors associated with concurrent stunting and overweight were investigated through interviewer administered questionnaires. A binary logistic regression model was used to determine the relation between nutritional statuses and associated factors. RESULTS: A low-income family and a low maternal educational level are independent risk factors for a child to be stunted-overweight [odds ratios (95% CI): 3.81 (13.32-1.08), 2.90 (7.82-1.07), respectively]. Mother under-evaluation of child's weight is a factor associated with stunting-overweight and overweight in children [odds ratios (95% CI): 3.42 (8.72-1.33), 6.52 (18.09-2.34), respectively]. Being overweight is also related to higher birth weight. Unlike stunted children, stunted-overweight children live with both their parents and have an older mother. Short maternal stature and mother's over-evaluation of her child's height are independent factors associated with stunting. CONCLUSION: Poor preschool children of Yaounde are facing concomitant stunting and overweight. More studies are needed to better assess the impact of maternal factors. PMID- 19191082 TI - Age-related risk of mortality from bladder cancer in diabetic patients: a 12-year follow-up of a national cohort in Taiwan. AB - BACKGROUND: To compare bladder cancer mortality between diabetic patients and the general population. MATERIALS AND METHODS: Yearly sex-specific mortality rates for age 25-64, 65-74, and > or =75 years in Taiwanese general population for 1995 2006 were calculated; 113,347 diabetic men and 131,573 diabetic women aged > or =25 years recruited in 1995-1998 were followed prospectively. RESULTS: In the general population, 4,943 men and 2,291 women died of bladder cancer, and aging was associated with increased risk. Although the trend of crude mortality was increasing in either sex, the trend of age-standardized rates had been steady. The average crude and age-standardized mortality rates were 5.35 and 5.98 (per 100,000 population), respectively, for men; and were 2.63 and 3.02 for women. A total of 224 diabetic men and 126 diabetic women died of bladder cancer with crude mortality of 26.0 and 11.9 per 100,000 person-years, respectively. The relative risk of bladder cancer mortality (95% confidence interval) for diabetic patients was 2.18 (1.75-2.72), 2.50 (2.06-3.04), and 5.95 (4.57-7.74), in men, and 1.34 (0.96-1.89), 2.48 (1.92-3.19), and 7.44 (5.46-10.15), in women, for ages > or =75, 65-74, and 25-64 years, respectively. CONCLUSIONS: Diabetic patients had a higher risk of bladder cancer mortality, which is more remarkable in the younger population. PMID- 19191083 TI - Marine antifouling laboratory bioassays: an overview of their diversity. AB - In aquatic environments, biofouling is a natural process of colonization of submerged surfaces, either living or artificial, involving a wide range of organisms from bacteria to invertebrates. Antifouling can be defined as preventing the attachment of organisms onto surfaces. This article reviews the laboratory bioassays that have been developed for studying the control of algae and invertebrates by epibiosis (chemical ecology) and the screening of new active compounds (natural products and biocides) to inhibit settlement or adhesion, ie fouling-release coatings. The assays utilize a range of organisms (mainly marine bacteria, diatoms, algae, barnacles). The main attributes of assays for micro- and macroorganisms are described in terms of their main characteristics and depending on the biological process assessed (growth, adhesion, toxicity, behavior). The validation of bioassays is also discussed. PMID- 19191084 TI - Antifouling activity of synthetic polymer gels against cyprids of the barnacle (Balanus amphitrite) in vitro. AB - Barnacle (Balanus amphitrite) settlement on synthetic hydrogels with various chemical structures was tested in laboratory assays. The results demonstrated that cyprids settle less or not at all on hydrogels and PDMS elastomer compared with the polystyrene control. The low settlement on gels is most likely due to the 'easy release' of initially attached cyprids from the gel surfaces. This low adhesion of cyprids is independent of surface hydrophilicity or hydrophobicity, and of surface charge. The results also revealed that hydrogels can be categorized into two groups. One group showed an extremely strong antifouling (AF) performance that was independent of the elasticity (E) or swelling degree (q) of the gels. The second group showed relatively less strong AF performance that was E- or q-dependent. In the latter case, E, rather than the q, may be the more important factor for cyprid settlement. PMID- 19191085 TI - ONTARGET, TRANSCEND and PROFESS--clarifying, confusing or misleading? PMID- 19191086 TI - Can 'partial' PTSD explain differences in diagnosis of PTSD by questionnaire self report and interview after head injury? AB - OBJECTIVE: Can the concept of 'partial' PTSD explain the disparity between the relatively high incidence of PTSD found using self-report questionnaires and the relatively low incidence using structured interview? It was hypothesized that self-report of greater PTSD symptom severity is associated with increased heart rate and movement when responding to questions about the traumatic event, if 'partial' PTSD is an explanation. RESEARCH DESIGN: A within participants single group design. METHODS: Twenty-one adults with head injury underwent self-report (Post-traumatic Stress Disorder Scale) and interview (Clinical Assessment of PTSD) assessments of PTSD, the Traumatic Memory Interview, self-report of mood (Hospital Anxiety and Depression Scale) and cognitive assessment (Wechsler Test of Adult Reading, Auditory Verbal Learning Test, Hayling Test, Digit Symbol Test), during which heart rate and motor activity were recorded. RESULTS: Self report of greater PTSD symptom severity was not associated with increases in heart rate or movement during questions about the traumatic event. In fact, heart rate decreased from baseline in those with higher self-report scores for PTSD, consistent with curiosity about the traumatic event and not 'partial' PTSD. CONCLUSION: These preliminary findings agree with an emerging theme suggesting that, although PTSD can occur after head injury, it is easily over-diagnosed. PMID- 19191087 TI - Neuropsychological functioning following complicated vs. uncomplicated mild traumatic brain injury. AB - OBJECTIVE: It would be logical to assume that patients with intracranial abnormalities (i.e. complicated MTBIs) would have worse outcome than patients without these abnormalities (i.e. uncomplicated MTBIs). However, the literature is limited and somewhat mixed regarding outcome in patients with complicated mild TBIs. The purpose of this study is to employ a carefully controlled research design to compare the acute neuropsychological functioning of patients following complicated and uncomplicated MTBI. METHOD: Participants were 20 patients with complicated MTBI and 20 patients with uncomplicated MTBI selected from an archival database of 465 patients. Patients were carefully matched on age, education, gender, ethnicity, days assessed post-injury and mechanism of injury. Patients were assessed an average of 3.5 days (SD = 1.9) post-injury with 13 common cognitive variables. RESULTS: There were significant group differences on only three of the 13 cognitive measures (complicated mild TBI worse than uncomplicated mild TBI). There were no significant differences in the proportion of impaired scores between groups on all measures, with the exception of Hopkins Verbal Learning Test Delayed Recall. CONCLUSION: Patients with complicated MTBIs performed more poorly only on a small number of tests during the acute recovery period. PMID- 19191088 TI - Social problem-solving abilities, relationship satisfaction and depression among family caregivers of stroke survivors. AB - PRIMARY OBJECTIVE: To examine the prospective relation of caregiver problem solving abilities and relationship satisfaction to caregiver depression 1 month following care recipient discharge from an inpatient stroke rehabilitation facility. RESEARCH DESIGN: Correlational, prospective design. METHODS AND PROCEDURES: Participants included 39 women (average age = 51.47, SD = 20.59) and four men (average age = 42.50, SD = 5.57) assuming caregiver roles for persons discharged from stroke rehabilitation. MAIN OUTCOME MEASURE: The Beck Depression Inventory was the outcome measure at discharge and 1 month later. RESULTS: Caregivers experienced a significant decrease in depression scores between the discharge and 1-month assessments. Lower relationship satisfaction and dysfunctional problem-solving abilities were independently and significantly predictive of depression scores at discharge. Dysfunctional problem-solving abilities were indirectly predictive of depression 1 month later. CONCLUSIONS: Dysfunctional problem-solving abilities and low relationship satisfaction are associated with caregiver depression, but dysfunctional problem-solving abilities maintain an indirect association with depression over 1 month. Implications of these findings for developing problem-solving training programmes for caregivers post-discharge are discussed. PMID- 19191090 TI - Impact of divided attention during verbal learning in young adults following mild traumatic brain injury. AB - PRIMARY OBJECTIVE: The goal of the present study was to assess the impact of mild traumatic brain injury (MTBI) on episodic memory performance in relation to attentional and executive control processes in young adults. RESEARCH DESIGN/METHODS: A verbal memory paradigm manipulating attentional load (full attention or divided attention) and semantic congruency between pairs of category target words during encoding was administrated to 13 individuals with MTBI and 12 normal control participants. Environmental supports during retrieval (free recall, cued recall and recognition modes) were also manipulated. MAIN OUTCOMES AND RESULTS: Results show that recall performances of individuals with MTBI were similar to those of controls when words were encoded under full attention. In contrast, individuals with MTBI performed worse than control participants when encoding under divided attention, whatever the semantic link between pairs of words. CONCLUSIONS: By using a sensitive test, one was able to objectively measure subtle impairments in memory performance, suggesting a diminished availability of attentional resources after MTBI. Young adults' learning of verbal material under divided attention might be compromised by the reduction of cognitive resources following MTBI. These findings are also discussed in light of different factors that can influence cognitive performance. PMID- 19191089 TI - Changes in responsiveness when brain injury survivors with impaired consciousness hear different voices. AB - PRIMARY OBJECTIVE: The researchers sought to determine whether individuals with impaired consciousness secondary to acquired brain injury (ABI) changed in responsiveness when purposefully presented with familiar, unfamiliar and synthetic voice messages. RESEARCH DESIGN: Researchers used an ABA single case study design across stimuli. Participants were three minimally-responsive ABI survivors. METHODS AND PROCEDURES: Participants heard auditory stimuli twice daily for 30 days. Data from video recordings included tallies of behavioural responses at 10-second intervals throughout baseline, intervention and post intervention phases of each session. Statistical calculations allowed determination of responsiveness changes across time intervals within sessions. MAIN OUTCOMES AND RESULTS: Unique response profiles emerged across survivors. Two participants demonstrated responsiveness changes with presentation of auditory stimuli. None demonstrated a clinically-significant differential response based on voice type. CONCLUSIONS: Findings suggest that auditory stimulation results in arousal changes in some ABI survivors, regardless of the familiarity of voices presented. PMID- 19191091 TI - Long-term effects of rivastigmine capsules in patients with traumatic brain injury. AB - OBJECTIVE: To investigate the safety, tolerability and efficacy of rivastigmine capsules (3-12 mg/day) in a 26-week, multi-centre, open-label extension of a double-blind study. METHODS: Patients with traumatic brain injury (TBI) and persistent cognitive impairment who had received rivastigmine (3-6 mg/day) or placebo for 12 weeks could enter the extension study and receive rivastigmine (< or =12 mg/day). Patients were assessed using a range of cognitive tests including the Hopkins Verbal Learning Test (HVLT) and the Cambridge Neuropsychological Test Automated Battery Rapid Visual Information Processing (CANTAB RVIP) A' sub-test. Safety measures included monitoring of adverse events. RESULTS: In the extension study (n = 127), the mean duration of rivastigmine treatment was 23.8 weeks and the mean final dosage was 7.9 mg/day. Approximately 40% of patients were responders (> or =1.0 SD improvement from baseline) on CANTAB RVIP A' or HVLT total score at week 38 or endpoint. Statistically significant changes from week 12 at week 38 were observed for CANTAB-RVIP A' and HVLT-total word recall for the sub-group of ex-placebo patients with greater severity of initial impairment. The safety profile of rivastigmine capsules was consistent with the label. CONCLUSIONS: Treatment with rivastigmine for up to 38 weeks was safe in patients with TBI and cognitive impairment. PMID- 19191092 TI - Kinematic analysis of articulatory coupling in acquired apraxia of speech post stroke. AB - PRIMARY OBJECTIVE: Electromagnetic articulography was employed to investigate the strength of articulatory coupling and hence the degree of functional movement independence between individual articulators in apraxia of speech (AOS). METHODS AND PROCEDURES: Tongue-tip, tongue-back and jaw movement was recorded from five speakers with AOS and a concomitant aphasia (M = 53.6 years; SD = 12.60) during /ta, sa, la, ka/ syllable repetitions, spoken at typical and fast rates of speech. Covariance values were calculated for each articulatory pair to gauge the strength of articulatory coupling. The results obtained for each of the participants with AOS were individually compared to those obtained by a control group (n = 12; M = 52.08 years; SD = 12.52). Comparisons were made between the typical rate productions of the control group and the typical and fast rate productions of the participants with AOS. MAIN OUTCOMES AND RESULTS: In comparison to the control group, four speakers with AOS exhibited significantly stronger articulatory coupling for alveolar and/or velar speech targets, during typical and/or fast rate conditions, suggesting decreased functional movement independence. CONCLUSIONS: The reduction in functional movement independence might have reflected an attempt to simplify articulatory control or a decrease in the ability to differentially control distinct articulatory regions. PMID- 19191093 TI - Scutellaria flavonoid supplementation reverses ageing-related cognitive impairment and neuronal changes in aged rats. AB - PRIMARY OBJECTIVE: To study the effects of flavonoid, isolated from aerial parts of Scutellaria baicalensis Georgi (SSF), on memory impairment, neuronal damage, free radicals and energy metabolite disorders in aged rats. METHODS AND PROCEDURES: Approximately 25-month-old rats were used to establish the ageing model. The cognition of the rats was determined using the Morris water maze, neuronal morphology was observed by light/electron microscope, malondialdehyde (MDA) levels and the activity of superoxide dismutase (SOD), lactate dehydrogenase (LDH) and Na(+)-K(+)-ATPase were measured by spectrophotometry. MAIN OUTCOMES AND RESULTS: In the Morris water maze task, the aged rats always took longer latency to find the hidden platform and spent less time swimming in the target quadrant than those of young control rats. The light/electron microscopic observations found significant neuropathological changes in the aged rats' brain. In addition, the production of MDA and the activity of SOD, LDH and Na(+)-K(+)-ATPase in the hippocampus and cerebral cortex of the aged rats showed critical abnormal changes. However, pre-treatment of the aged rats with SSF (35 140 mg kg(-1)) for 16-21 days dramatically improved cognitive dysfunction, neuropathological changes and biochemical abnormalities. CONCLUSION: These results indicate that the beneficial effects of SSF on memory impairment and neuronal damage in aged rats may be important for the treatment of senile dementia and for delaying the ageing processes. PMID- 19191094 TI - A learning assessment procedure to re-evaluate three persons with a diagnosis of post-coma vegetative state and pervasive motor impairment. AB - PRIMARY OBJECTIVE: Detecting signs of learning in persons with a diagnosis of post-coma vegetative state and profound motor disabilities could modify their diagnostic label and provide new hopes. In this study, three adults with such a diagnosis were exposed to learning assessment to search for those signs. PROCEDURE AND DESIGN: The assessment procedure relied on participants' eye blinking responses and microswitch-based technology. The technology consisted of an electronically regulated optic microswitch mounted on an eyeglasses' frame that the participants wore during the study and an electronic control system connected to stimulus sources. Each participant followed an ABABCB design, in which A represented baseline periods, B intervention periods with stimuli contingent on the responses and C a control condition with stimuli presented non contingently. MAIN OUTCOMES AND RESULTS: The level of responding during the B phases was significantly higher than the levels observed during the A phases as well as the C phase for all participants (i.e. indicating clear signs of learning by them). CONCLUSIONS: These findings may have important implications for (a) changing the participants' diagnostic label and offering them new programme opportunities and (b) including learning assessment within the evaluation package used for persons with post-coma profound multiple disabilities. PMID- 19191095 TI - Late recovery from post-traumatic vegetative state. AB - AIM: Late recovery of consciousness after 12 months in vegetative state is very rare and is almost invariably associated with severe disability. The aim of this study was to report that late recovery with moderate disability is possible, even after several months in vegetative state. CASE REPORT: This study describes the case of a 22-year-old male student who recovered consciousness from vegetative state 19 months after a traumatic brain injury which had occurred in December 2000. A further slow recovery of motor and cognitive functions up to a moderate disability was observed over 7 years after the brain injury. The patient now lives in the community, speaks, walks and undertakes daily living as well as outdoor leisure activities independently, has resumed his previous university studies and has a part-time non-competitive job. CONCLUSIONS: This case demonstrates that attaining a condition of independent living is possible even after more than 1 year of post-traumatic vegetative state, with an improvement of motor and cognitive functions that can continue for years. Cranioplasty and long term rehabilitation programmes may be among the variables potentially influencing this unexpected recovery. PMID- 19191096 TI - Language and cognition in a bilingual child after traumatic brain injury in infancy: long-term plasticity and vulnerability. AB - PRIMARY OBJECTIVE: This study aimed at investigating the long-term effects of the combination of severity of injury and time of injury in a 6-year-old bilingual Arabic-Italian child who sustained a severe left traumatic brain injury at the age of 7 months. METHODS AND PROCEDURES: Standard neurological, cognitive and neuropsychological assessments were administered at 40 days after surgery and again at 18, 31, 62 and 73 months. MAIN OUTCOMES AND RESULTS: The child presented with developmental arrest at 18 and 31 months. Later on, right hemiparetic and oculomotor signs gradually improved to a significant extent, as well as dysexecutive, visuospatial and praxic deficits. At present, persistent language disorders in a fluent speech characterize the child's profile to a similar extent and type in both languages, suggesting common underlying learning strategies which are ineffective for procedurally acquiring language. CONCLUSIONS: This case confirms that children who sustain severe left hemisphere traumatic brain injury in infancy present with increased vulnerability to linguistic deficits. Left frontotemporal, cortical-subcortical lesions which occur during very early language development may permanently disrupt the procedural language acquisition network required for first language acquisition. PMID- 19191097 TI - Fast awakening from minimally conscious state with apomorphine. AB - BACKGROUND: Traumatic brain injury (TBI) can induce long-term severe disorders of consciousness. Evidence suggests an underlying dopaminergic deficit. Dopamine agonists may therefore play an important role in recovery of consciousness. OBJECTIVE: To explore the response to continuous subcutaneous administration of apomorphine in a patient who had remained in minimally conscious state for 104 days and to evaluate the anatomical substrate of the effect. DESIGN: A prospective, open-label, daily treatment, dose-escalation single case clinical study, with retrospective diffusion tensor image (DTI) evaluation. RESULTS: On the fist day of treatment, the patient was able to move his limbs on command and answer yes/no questions which had not been the case prior to apomorphine administration. Subsequently there was a full recovery of consciousness and substantial functional recovery that was sustained even after apomorphine discontinuation. At the highest dose, mild dyskinesias were observed. These resolved with a lowering of the dose. DTI demonstrated a decrease of thalamocortical and corticothalamic projections in this MCS patient compared to normal volunteers. CONCLUSION: Although this is an open-label single-patient case report, the data are consistent with the theory that a dopaminergic deficit underlies MCS and that it may be overcome with apomorphine administration. PMID- 19191098 TI - Development and validation of gene expression profile signatures in early-stage breast cancer. PMID- 19191099 TI - Metastatic breast cancer: overall survival related to successive chemotherapies. What do we gain after the third line? AB - This retrospective study analyzed, in metastatic breast cancer (MBC), overall survival (OS) related to each subsequent line of chemotherapy (CT). We evaluated 578 patients with a MBC. Among these patients, 558 patients were stratified according to the number of different CT lines received. Median OS decreased from 22.5 to 12.3 months. Nevertheless, survival was rather stable for subsequent lines after the third line of CT around 8 months. In our study, it appeared useful to give several CT lines when possible. Subsequent lines of CT could have a beneficial effect on the survival for some patients with MBC. PMID- 19191100 TI - Informatics or information technology for cancer research: which is it? PMID- 19191101 TI - Gender difference in access and intervention outcomes: the case for women with disabilities. AB - PURPOSE: This study replicated the one that was conducted 30 years ago to determine systematic gender differences in access and outcomes of vocational rehabilitation services (VRS). METHODS: Unlike the original study that was confined to Region V, this study examined case management data from 653,206 cases that were closed by State VRS in 2004 in all states. It used a t-test to analyse gender differences in access to VRS and intervention outcomes in education, employment, earnings and dependence on public support. The study also conducted multivariate analysis to determine the effects of gender on VRS intervention outcomes. RESULTS: The study found differences similar to those found 30 years ago. Although women had better educational outcomes, they had lower employment and earnings and greater dependence on needs-based public support than did men. A disturbing finding is that the gender employment and earnings gaps were wider after VRS interventions than before. After controlling for other factors, the study found that gender is a significant predictor of earnings and employment outcomes. CONCLUSION: These findings suggest that men with disabilities stand a better chance of escaping poverty than do women. There is a need for changes in VRS service provision that ensures equity in quality of services which might translate into more equitable employment and earnings outcomes. PMID- 19191102 TI - Inhibition of interleukin-10 signaling in lung dendritic cells by toll-like receptor 4 ligands. AB - The homeostatic microenvironment in lung is immunosuppressive and interleukin-10 (IL-10) helps maintain this microenvironment. Despite constitutive production of IL-10 in normal lung, macrophages (MOs) and dentritic cells (DCs) remain capable of responding to microorganisms, suggesting that these innate immune cells have a mechanism to override the immunosuppressive effects of IL-10. Prior studies by the authors revealed that Toll-like receptor (TLR) ligands inhibit IL-10 receptor signaling in alveolar macrophages (AMOs), thereby obviating the immunosuppressive activity of IL-10. This report compares the immunologic phenotypes of AMOs and lung DCs and their ability to respond to IL-10 following exposure to microbial stimuli. IL-10 was constitutively produced by normal lung epithelium and exposure to lipopolysaccharide (LPS) in vivo increased the expression of IL-10 during the first 24 hours. AMOs constitutively produced IL-10 mRNA, whereas both AMOs and LDCs constitutively expressed IL-12 mRNA. AMOs and LDCs, as well as bone marrow derived MOs and DCs, had reduced capacity to activate STAT3 in response to IL-10 if pretreated with LPS. Inhibition was not associated with decreased expression of IL-10 receptor (IL-10R) and was dependent on the MyD88 signaling pathway. These results demonstrate a common underlying regulatory mechanism in both DCs and MOs by which microbial stimuli can override the immunosuppressive effect of constitutive IL-10 production in the lung. PMID- 19191103 TI - ErbB4 regulates surfactant synthesis and proliferation in adult rat pulmonary epithelial cells. AB - ErbB4 is a predominant heterodimer for other ErbB receptors in late fetal lung development where it participates in regulating type II cell surfactant synthesis. To further elucidate the role of ErbB4 in pulmonary alveolar epithelial cell function, the authors hypothesized that ErbB4 participates in maintaining adult lung type II cell homeostasis. The authors used small interfering RNA (siRNA) to down-regulate endogenous, ErbB4 receptors in the adult rat lung epithelial L2 cell line and measured neuregulin 1beta (NRG1beta)-, and fibroblast conditioned medium (FCM)-induced effects on L2 cell surfactant phospholipid synthesis and proliferation. Under control conditions, total and phosphorylated ErbB4 were significantly increased after both NRG1beta and FCM treatment, as were surfactant phospholipids synthesis and cell proliferation. Down-regulation of ErbB4 with siRNA reduced stimulation of NRG1beta- and FCM induced ErbB4 phosphorylation, decreased endogenous surfactant phospholipid synthesis, and blocked NRG1beta- and FCM-stimulated surfactant phospholipid synthesis. NRG1beta- and FCM-induced cell proliferation was not affected. The authors conclude that ErbB4 participates in maintaining adult lung alveolar epithelial cell surfactant synthesis and proliferation with development-specific functions. PMID- 19191104 TI - C-reactive protein modulates human lung fibroblast migration. AB - C-reactive protein (CRP) has been classically used as a marker of inflammation. The aim of this study was to investigate the effect of CRP on migration of human fetal lung fibroblasts (HFL-1) to human plasma fibronectin (HFn). Using the blindwell chamber technique, CRP inhibited HFL-1 migration in a dose-dependent fashion (at 1 microg/mL, inhibition: 32.5% +/- 7.1%; P < .05). Western blot analysis showed that CRP inhibited the p38 mitogen-activated protein kinase (MAPK) activity in the presence of HFn. Moreover, the MAPK inhibitors SB202190 (25 microM) and SB203580 (25 microM) inhibited HFn-induced cell migration, suggesting an important role of p38 MAPK in HFn-induced migration. Taken together, these results suggest that the inhibitory effect of CRP is mediated by blocking MAPK. In summary, this study demonstrates that CRP directly modulates human lung fibroblasts migration. Thus, CRP may contribute to regulation of wound healing and may be endogenous antifibrotic factor acting on lung fibrosis. PMID- 19191105 TI - Nuclear localization of active matrix metalloproteinase-2 in cigarette smoke exposed apoptotic endothelial cells. AB - Cigarette smoke (CS)-induced activation of proteases such as matrix metalloproteinases (MMPs) contributes to lung alveolar destruction due to cell death. The aim of this study was to determine whether MMPs are produced in pulmonary artery endothelial cells (PAECs) and whether CS activation of MMPs is associated with apoptosis. Cultured PAECs were exposed to CS and subjected to assessments of apoptosis and MMPs. Western blotting and in situ zymography were performed to localize gelatinolytic activity and to identify enzymes. CS-induced apoptosis, i.e., enhanced annexin V binding and cleaved poly-ADP-ribose polymerase (PARP), correlated with increased degradation of gelatin, a substrate of MMPs. The levels of pro-MMP-2 and active MMP-2 were increased in cytosolic and nuclear fractions isolated from CS-exposed cells. MMP-2 protein colocalized with gelatinolytic activity in the nucleus of CS-exposed cells undergoing apoptosis. These observations support the notion that MMP-2 contributes to CS-induced gelatinase activity, which localizes in the nuclear region primarily and correlates with annexin V binding and PARP cleavage. This suggests a novel function of MMP-2 in the degradation of the nuclear matrix in CS-induced endothelial apoptosis. PMID- 19191106 TI - Polyethylene glycol addition does not improve exogenous surfactant function in an experimental model of meconium aspiration syndrome. AB - Meconium (MEC) is a potent inactivator of pulmonary surfactant. The authors studied the effects of polyethylene glycol addition to the exogenous surfactant over the lung mechanics and volumes. Human meconium was administrated to newborn rabbits. Animals were ventilated for 20 minutes and dynamic compliance, ventilatory pressure, and tidal volume were recorded. Animals were randomized into 3 study groups: MEC group (without surfactant therapy); S100 group (100 mg/kg surfactant); and PEG group (100 mg/kg porcine surfactant plus 5% PEG). After ventilation, a pulmonary pressure-volume curve was built. Histological analysis was carried out to calculate the mean alveolar size (Lm) and the distortion index (DI). Both groups treated with surfactant showed higher values of dynamic pulmonary compliance and lower ventilatory pressure, compared with the MEC group (P < .05). S100 group had a larger maximum lung volume, V(30), compared with the MEC group (P < .05). Lm and DI values were smaller in the groups treated with surfactant than in the MEC group (P < .05). No differences were observed between the S100 and PEG groups. Animals treated with surfactant showed significant improvement in pulmonary function as compared to nontreated animals. PEG added to exogenous surfactant did not improve lung mechanics or volumes. PMID- 19191107 TI - Protection of pyrroloquinoline quinone against methylmercury-induced neurotoxicity via reducing oxidative stress. AB - Pyrroloquinoline quinone (PQQ) is a novel redox cofactor and also exists in various foods. In vivo as well as in vitro experimental studies have shown that PQQ functions as an essential nutrient or antioxidant. Methylmercury (MeHg), as a highly toxic environmental pollutant, could elicit central nervous system (CNS) damage. Considering the antioxidant properties of PQQ, this study was aimed to evaluate the effect of PQQ on MeHg-induced neurotoxicity in the PC12 cells. The results showed that, after pre-treatment of PC12 cells with PQQ prior to MeHg exposure, the MeHg-induced cytotoxicity was significantly attenuated and then the percentage of apoptotic cells and the arrest of S-phase in cell cycle were correspondingly reduced. Moreover, PQQ significantly decreased the production of ROS, suppressed the lipid peroxidation and increased the antioxidant enzyme activities in PC12 cells exposed to MeHg. These observations highlighted the potential of PQQ in offering protection against MeHg-induced neuronal toxicity. PMID- 19191108 TI - Serotonin binds to purified neuronal nitric oxide synthase: a possible explanation for ROS production induced by 5HT in the presence of nNOS. AB - Serotonin (5HT) was shown to induce in vitro the production of ROS in the presence of neuronal nitric oxide synthase (nNOS) in addition to the basal NO(+) formation. With the aim of understanding this mechanism, this study investigated the potential binding of 5HT to nNOS. By using [(3)H]5HT, it is reported here that 5HT binds to nNOS, but only when the enzyme is active and in a superoxide dependent manner. This binding is prevented by DPI but not by L-NAME. The formation of 5HT-nNOS complex was shown to be very well correlated with the production of ROS by 5HT in the presence of nNOS. A mechanism involving nNOS only in its initial step is proposed to explain both the formation of 5HT-nNOS complex and the production of ROS observed in the presence of nNOS and 5HT. PMID- 19191109 TI - Vascular pro-oxidant effects related to the autoxidation of dopamine. AB - Dopamine autoxidation in an oxygenated physiological salt solution (37 degrees C, pH=7.4) mostly occurred in a 2.5 h time period. H(2)O(2) and dopamine quinones were produced during dopamine autoxidation. In partially pre-contracted rat aortic rings, 10-100 microm dopamine induced endothelium-independent contractions and 0.3-1 mm dopamine induced complete, slow-developing endothelium-independent relaxations. Indomethacin and catalase suppressed the endothelium-independent dopamine contractions. Catalase strongly reduced the endothelium-independent dopamine relaxations. Furthermore, 1 mm dopamine for 60 min followed by a 90 min washout period induced the release of lactate dehydrogenase and the complete impairment of ring reactivity to phenylephrine and KCl. Pre-treatment with catalase or glutathione prevented dopamine-induced deleterious effects so that further concentration-response curves to phenylephrine and KCl could be obtained. The phenylephrine potency was maintained in rings pre-treated with glutathione but not in rings pre-treated with catalase. IN CONCLUSION: (1) dopamine is rapidly and non-enzymatically oxidized in physiological solutions, generating H(2)O(2) and quinones; (2) low H(2)O(2) levels increase vascular tone by activating cyclooxygenase; (3) high H(2)O(2) levels cause irreversible relaxations due to unspecific cellular damage; and (4) dopamine quinones cause a specific alteration in the phenylephrine response. PMID- 19191110 TI - Osmotic stress induces loss of glutathione and increases the sensitivity to oxidative stress in H9c2 cardiac myocytes. AB - It has been observed that H9c2 cardiac cells cultured in physiologic solutions exhibit delayed cell death after repeated medium replacements, of which the cause was the relatively mild osmotic challenges during the renewal of the culture medium. Interestingly, the cell damage was associated with altered intracellular GSH homeostasis. Therefore, this study attempted to elucidate the effects of osmotic stress on GSH metabolism. In cells subjected to osmotic stress by lowering the NaCl concentration of the medium, the cell swelling was rapidly counterbalanced, but the intracellular GSH content was significantly lower in 3 h. Meanwhile, the ratio of GSH-to-GSSG was not affected. As expected, osmotic stress also increased the sensitivity to H(2)O(2), which was attributable to the decrease of GSH content. The decrease of GSH content was similarly evident when the synthetic pathways of GSH were blocked by BSO or acivicin. It was concluded that osmotic stress induced the decrease of intracellular GSH content by increased consumption and this loss of GSH rendered the cells susceptible to a subsequent oxidative stress. PMID- 19191111 TI - Editorial--ICOWHI special issue. PMID- 19191112 TI - Voices from the frontlines: the epidemics of HIV/AIDS and violence among women and girls. PMID- 19191113 TI - A global imperative: development, safety, and health from girl child to woman. AB - The purposes of this article are to review what we know about the health status for girl children in the world and to suggest policy that allows us to see how to improve the health of girl-children. First, I will consider globalization and its relationship to health disparities. I then will review what we know about the health of the girl-child, contrasting patterns of health observed in girl children from Sub-Saharan Africa and the consequences for subsequent generations. Finally, I will focus on interventions at the level of social and health policy, the community, and systems of health care delivery. PMID- 19191114 TI - Remarks by His Excellency Mr. Festus G. Mogae, former president of the Republic of Botswana: accepting the Taylor and Francis Award. PMID- 19191115 TI - Speech by Gogontlejang Phaladi: the 17th ICOWHI Congress 2008--Wednesday, 9 July 2008. PMID- 19191116 TI - Disparity in access to cataract surgical services leads to higher prevalence of blindness in women as compared with men: results of a national survey of visual impairment. AB - AIM: The aim of the survey was to estimate the prevalence and determinants of visual impairment and blindness in Botswana 50 years and older and assess access to cataract surgical services. METHOD: a multistage cluster randomized sampling with probability proportional to size was used to select a cross-sectional nationally representative sample of 2,662 subjects. The distance visual acuity was measured and the lens examined for cataracts in each subject. Where pinholes did not improve vision to 6/18 or better, fundus examination was done after mydriasis. Blindness was defined as vision <3/60 and visual impairment as <6/18 to 3/60 in the better eye with available correction. RESULTS: Two thousand one hundred twenty-seven eligible subjects were examined (79.9%). Age- and sex adjusted prevalence of blindness and severe visual impairment was 3.69% (95% CI: 2.38%-5.00%) and 1.69% (95% CI: 1.04%-2.33%), respectively. Cataracts were the main cause of blindness (46.9%), and severe (58.9%) and moderate (40.2%) visual impairment. After adjusting for age and sex, the prevalence of bilateral cataract blindness and bilateral severe visual impairment in men is 1.0% (95% CI: 0.001% 2.1%) and 1.5% (95% CI: 1.3%-1.7%) compared with 1.6% (95% CI: 0.6%-2.7%) and 2.1% (95% CI: 1.8%-2.8%) in women. 76.9% of men, bilaterally blind from cataract, had cataract surgery in one or both eyes compared with 59.4% of women. Overall, 65.4% of people bilaterally blind from cataract had surgery in one or both eyes. For vision <6/60, the cataract surgical coverage (CSC) for persons is 73.0%, 55.2%, and 61.5% for men, women, and the sample population, respectively. The CSC for persons at vision <6/18 is 62.3%, 48.3%, and 53.2% for men, women, and the sample population, respectively. CONCLUSION: Inequity in access to cataract surgery between men and women in Botswana has contributed to the higher prevalence of cataract-related blindness in women. PMID- 19191117 TI - Intimate partner violence and sexually risky behavior in Botswana: implications for HIV prevention. AB - AIM: Evidence-based research has identified a link between experiencing intimate partner violence (IPV) and exposure to HIV in the general population and for women and girls in sub-Saharan Africa in particular. Emerging evidence in Sub Saharan Africa supports the fact that violence perpetrated against women and girls tends to increase their risk of HIV infection. The aim of the study was to explore the type and severity of IPV and HIV-related risk behaviors among remote, rural, and urban dwellers in Botswana aged 15 and over. METHODS: The study combined both quantitative and qualitative methods of inquiry. Respondents came from remote area settlements and rural and urban areas of Botswana. The sample population was 1,378 Botswana, who are 15 years and above. Face-to-face structured and unstructured interviews were conducted by trained interviewers. RESULTS: Thirty-two percent of the respondents reported that they had been in a relationship that they perceived as abusive. Respondents had had at least 2.4 abusive relationships. The mean Habitat Sustainability Index summed score was 2.4 (SD = 2.2). Women were more likely to report having been in an abusive relationship than men. The linear combination of physical, verbal, emotional, and sexual abuse explained 37% of the variance in sexually risky behaviors, r(2) = .37, F (13, 113) = 4.113, p <. 001, F change = 4.392. Emotional abuse and sexual abuse were the best predictors of sexually risky behaviors. When categories of abuse were classified by physical and nonphysical abuse, nonphysical (verbal and emotional abuse) accounted for a greater part of the variance in sexual high-risk behavior (beta = .32, p = .001). CONCLUSION: Intimate partner violence (IPV) is common in Botswana and is related to high-risk sexual behavior. Programs for the prevention of HIV should adopt a life course model in order to identify the underlying factors in high-risk behavior. PMID- 19191118 TI - Risks and protective behaviors of adolescents living with HIV/AIDS in Botswana. AB - HIV and AIDS are a major problem in Botswana. Studies have revealed that they have mostly affected young people, most of whom will have been infected with HIV during their teenage years. The prevalence among young people aged 15-24 is 16.9% and 6.6% among adolescents aged 15-19 years. It is therefore crucial to examine perceived contextual factors that influence adolescents living with HIV and AIDS, the risks that they encounter, and the behaviors that they use to reduce such risks. A qualitative exploratory descriptive design will be used to describe the HIV and AIDS knowledge risk and protective factors in general and sexual factors in particular of adolescents living with HIV and AIDS. Focus group discussions and individual in-depth interviews will be conducted among female and male adolescents aged 13 to 19 years. Content analysis will be utilized. PMID- 19191119 TI - People living with HIV and AIDS on the brink: stigma--a complex sociocultural impediment in the fight against HIV and AIDS in Botswana. AB - HIV-related stigma is a life-altering phenomenon. The consequence of the stigmatization process sets apart stigmatized person(s) as a distinct category, leading to various forms of disapproval, rejection, exclusion, labeling, stereotyping, and discrimination. Stigma of HIV-positive people in Botswana is a complex social phenomenon associated with the disease itself and the behaviors that lead to infection. This is a synthesis paper based on the literature review on HIV- and AIDS-related stigmatization of HIV-positive people in Botswana and in depth interviews with people living with HIV and AIDS (PLWHAs). I examine the literature on HIV- and AIDS-related stigmatization and subsequent discrimination and the implications for intervention programs for people living with HIV and AIDS. The findings from the literature and in-depth interviews show that HIV-AIDS related stigma is deeply embedded in societal structures and culture which promote nonacceptance of those branded HIV positive. This often is reinforced at a practical level by pervasive negative attitudes toward PLWHAs. Recommendations argue for the adoption of Healthy Relationship. This intervention seeks to promote and strengthen decision-making skills among PLWHAs and programs that promote destigmatization of, and tolerant attitudes toward, PLWHAs. PMID- 19191120 TI - Outcomes evaluation of St Petersburg Russia Women's Wellness Center. AB - Researchers demonstrate that investing in women's health yields great social and economic dividends, yet the health of women often receives little attention throughout the world. Women's wellness centers (WWCs) were established in the former Soviet Union beginning in 1992 under the sponsorship of the American International Health Alliance (AIHA) in conjunction with the United States Association for International Development (USAID) to provide a women-centered approach to primary care, providing access to a wide range of clinical services and educational outreach programs. Because the St Petersburg Women's Wellness Center (WWC) is considered one of the more advanced centers and represents a microcosm of opportunities and challenges in women's health care delivery in the Russian Federation, a descriptive qualitative outcomes evaluation of the clinic was conducted. In the spirit of continuous quality improvement, interviews were conducted with childbearing women. Findings are categorized into the dimensions of access, privacy and comfort, communication with providers, comprehensiveness of care, and follow-up care. Strengths of the clinic include the "woman friendly" environment and educational offerings. Suggestions also were made by study participants for improvement of clinic services. Qualitative research can be translated into clinical practice guidelines for WWCs in Eastern Europe. PMID- 19191121 TI - Trapped in circle of threats: violence against sex workers in Thailand. AB - Most researchers studying sex work have focused on the risks of sexually transmitted diseases, predominantly HIV, for sex workers, their clients, and subsequent partners. Violence against these women often goes undocumented and unnoticed. Consequently, few researchers have addressed violence against sex workers, and these few have generated limited evidence about the nature of violence from the sex-worker perspective--especially the street sex worker perspective. In this study, we used qualitative methods to explore characteristics of violence against street sex workers and how violence influences personal and societal health risks. The participants were 28 female street sex workers. The data were collected through in-depth interviews with 23 women, one focus group with 5 women, and observations of these women in their working and social environment. The results revealed that violence against sex workers can be clustered into three categories, threat to their life and health, threat to control of work and financial security, and finally, threat to humanity. Because they are disadvantaged, and engage in illegal employment, theses women were trapped in a circle of threats. To reduce violence, sex work should be decriminalized along with strategies to decrease poverty and social inequality. A special agency needs to be established to protect the rights and safety of sex workers. PMID- 19191122 TI - Correspondence between human diet, body composition and stable isotopic composition of hair and breath in Fijian villagers. AB - The main aim of this work was to describe the relationship between diet, and hair and breath isotopic composition. From one Fijian rural village, hair and breath samples were procured from 20 women. Physical anthropometrics were made, and hair (13)C/(12)C and (15)N/(14)N and breath (13)C/(12)C were measured. Individual diet diaries were kept for two of the four preceding weeks, and isotopic compositions of items which accounted for most of the diet were measured. Individual average diets were analysed for macronutrient and energy content and conform to reasonable nutritional expectation. Characteristics of the diet are described in terms of protein and energy, their patterning with respect to different clusters of food items and their relationship to individuals' anthropometry. Breath CO(2) is depleted in (13)C by 1-2 per thousand on average with respect to the total diet. Hair was enriched on average by 4.1 per thousand in nitrogen and 4.5 per thousand in carbon with respect to the total diet. There was insufficient population variation in hair isotopic composition to establish individual hair diet isotopic differences. The definite relationship that we establish in this work, between dietary and tissue isotopic values for a human community, provides a basis for determining and validating dietary regimes more generally within non industrial, non-global-'supermarket' economies. PMID- 19191123 TI - Effect of molecule size on carbon isotope fractionation during biodegradation of chlorinated alkanes by Xanthobacter autotrophicus GJ10. AB - The effect of the number of carbon and chlorine atoms on carbon isotope fractionation during dechlorination of chlorinated alkanes by Xanthobacter autotrophicus GJ10 was studied using pure culture and cell-free extract experiments. The magnitude of carbon isotope fractionation decreased with increasing carbon number. The decrease can be explained by an increasing probability that the heavy isotope is located at a non-reacting position for increasing molecule size. The isotope data were corrected for the number of carbons as well as the number of reactive sites to obtain reacting-site-specific values denoted as apparent kinetic isotope effect (AKIE). Even after the correction, the obtained AKIE values varied (on average 1.0608, 1.0477, 1.0616, and 1.0555 for 1,2-dichloroethane, chloropentane, 1,3-dichloropentane and chlorobutane, respectively). Cell-free extract experiments were carried out to evaluate the effect of transport across the cell membrane on the observed variability in the AKIE values, which revealed that variability still persisted. The study demonstrates that even after differences related to the carbon number and structure of the molecule are taken into account, there still remain differences in AKIE values even for compounds that are degraded by the same pure culture and an identical reaction mechanism. PMID- 19191124 TI - Isotopic composition of nitrogenous compounds with emphasis on anthropogenic loading in river ecosystems. AB - Nitrogenous compounds with high delta(15)N values were recently found in human dominated small rivers in the Lake Biwa area. A detailed survey was performed to determine the distribution and variation of delta(15)N values in nitrogenous compounds in a representative small river (Hebisuna River) that flows into Lake Nishinoko, an inner bay of Lake Biwa. A high delta(15)N value was detected in the lower reaches of the river and the inner bay, most likely due to denitrification. These results strongly suggest that denitrification in small river systems such as the Hebisuna watershed has contributed to (15)N enrichment in the Lake Biwa ecosystem during the past 40 years. We also observed a clear, stepwise, positive correlation between population density and delta(15)N values for particulate organic matter or sediments. These results demonstrate that delta(15)N (POM) and delta(15)N (sediment) are helpful indicators for assessing nitrogen loading from domestic sewage. Moreover, they will aid in the development of new concepts in the environmental capacity of river ecosystems and its relationship to redox conditions. Finally, our data suggest that a population density of 100-200 persons per km(2) is the upper limit for a watershed in which only simple conventional sewage treatment is in effect. PMID- 19191125 TI - Isotopic evidence of the effects of herbivory and landscape position on plant nitrogen sources in a riparian ecosystem. AB - This study used (15)N natural abundance techniques to investigate whether elk herbivory affects the capability of willow (Salix spp.) and reed-grass (Carex spp.) to access N from groundwater in upper and lower landscape positions within riparian areas of the Rocky Mountain National Park. For this, delta(15)N values of plant, soil, rainfall and groundwater samples were measured within a replicated field experiment. Despite the relatively high variability in the (15)N data, it provided evidence of the effects of grazing and lanscape position on plant N sources. Carex seemed to acquire less groundwater N in upper landscape positions compared to Salix, maybe due to the deeper willow rooting system. However, grazed willows in upper landscape positions seemed to acquire less groundwater N as compared to willows protected from herbivory. These findings were in accordance with other isotopic and non-isotopic studies conducted in the same experimental site. PMID- 19191126 TI - Headspace solid phase microextraction--GC/C-IRMS for delta13CVPDB measurements of mono-aromatic hydrocarbons using EA-IRMS calibration. AB - This work aims at comparing the delta(13)C(VPDB) of mono-aromatic hydrocarbons benzene, toluene, ethylbenzene and xylene isomers (BTEX) measured by elemental analyser (EA)-isotope ratio mass spectrometer (IRMS) with the delta(13)C(VPDB) measured on the same compounds by headspace solid phase microextraction - GC/C IRMS (hSPME - GC/C-IRMS) with the final goal of using these compounds as internal standards on the latter system. The EA-IRMS measurements were done using calcium and lithium carbonate isotopic reference materials: NBS19 and L-SVEC for establishing the delta(13)C(VPDB) scale. The EA-IRMS measurements with helium dilution of a set of five reference materials (USGS40, USGS41, IAEA-CH-6, IAEA-CH 3 and IAEA-601) show systematic bias of 1 per thousand relative to their assigned values. This bias due to the dilution mechanism in the used ConfloII interface device could not be avoided. As the selected hydrocarbons: BTEX could not be analysed by EA-IRMS without helium dilution, their delta(13)C(VPDB) must be corrected from this observed bias using an external calibration. The CO(2) gas calibrated using EA-IRMS without helium dilution, was used as an in-house reference for the delta(13)C(VPDB) measurements of the BTEX by the hSPME - GC/C IRMS system. The comparison made between the delta(13)C(VPDB) measured on the same BTEX compounds by EA-IRMS (with external calibration) and by hSPME - GC/C IRMS techniques showed good agreement. PMID- 19191127 TI - Laser-induced fluorescence spectroscopy of 14N18O and its application to breath analysis. AB - Laser-induced fluorescence (LIF) spectroscopy is an efficient tool for the detection of low nitric oxide (NO) concentrations down to the parts per trillion (ppt) range. The isotopic selectivity of this method provides a broad potential of applications. Whereas (14)N(16)O and (15)N(16)O have been described extensively this is not the case for the (14)N(18)O isotopologue. A (1+1) resonance enhanced multiphoton excitation time-of-flight mass spectrometer has been used for the simultaneous detection of (14)N(16)O, (15)N(16)O and (14)N(18)O isotopologues. Regions with single (14)N(18)O transitions have been studied in more detail by the LIF device. Electronic excitation of the A-X transition in the UV provides fluorescence which is monitored around 247 nm from A(2)Sigma(+)(v'=0) ->X(2)Pi(Omega)(v''=2). This transition has been used for an isotope specific online detection of (14)N(18)O from exhaled air. The detection limit for (14)N(18)O is 7.3+/-1.8 ppt. With respect to (18)O-labelled drugs and amino acids this denotes a novel extension to NO research. PMID- 19191128 TI - A new serial pooling method of shifted tree ring blocks to construct millennia long tree ring isotope chronologies with annual resolution. AB - The study presents a new serial pooling method of shifted tree ring blocks for the building of isotope chronologies. This method combines the advantages of traditional 'serial' and 'intertree' pooling, and can be recommended for the construction of sub-regional long isotope chronologies with sufficient replication, and on annual resolution, especially for the case of extremely narrow tree rings. For Scots pines (Pinus sylvestris L., Khibiny Low Mountains, NW Russia) and Silver firs (Abies alba Mill., Franconia, Southern Germany), serial pooling of five consecutive tree rings seems appropriate because the species- and site-specific particularities lead to blurs of climate linkages in their tree rings for the period up to ca. five years back. An equivalent to a five-year running means that curve gained on the base annual data sets of single trees can be derived from the analysis of yearly shifted five-year blocks of consecutive tree rings, and therefore, with approximately 20% of the expense. Good coherence of delta(13)C- and delta(18)O-values between calculated means of annual total rings or late wood data and means of five-year blocks of consecutive total tree rings analysed experimentally on most similar material confirms this assumption. PMID- 19191129 TI - Alveolar macrophage subpopulations in bronchoalveolar lavage and induced sputum of asthmatic and control subjects. AB - BACKGROUND: Alveolar macrophages (AM) are the most numerous immune cells in the airways and are involved in the immunological homeostasis of the lung. Intriguingly, their role in asthma remains unclear probably, in part, because of their heterogeneity. OBJECTIVE: To characterize AM population from bronchoalveolar lavage (BAL) and induced sputum (IS) of asthmatic and normal subjects using specific biomarkers. METHODS: Non-asthmatic non-allergic and allergic mild asthmatic subjects were recruited for this study. AM were obtained from BAL and IS and cytospins were prepared. Immunocytochemistry was performed for nine cellular markers (CD68, RFD7, CD14, CD11b, CD83, CD64, CD80, CD86, and FIZZ1). RESULTS: Asthmatic subjects had more AM RFD7(+) in BAL compared with IS, whereas control subjects had more AM RFD7(+) in IS than in BAL. Consequently, there was an increased number of AM RFD7(+) in BAL of asthmatic subjects compared with BAL of control subjects. AM CD11b(+) was higher in BAL than in IS in both groups. The expression of FIZZ1, marker of macrophage alternative activation, was similar in asthmatic and normal subjects. CONCLUSION: The expression of cellular markers on AM differs according to their localization in the lung. Subpopulations of AM may contribute to the inflammatory profile observed in asthmatic subjects. PMID- 19191130 TI - The association of BMI and sedentary time with respiratory symptoms and asthma in 5th grade schoolchildren in Kaohsiung, Taiwan. AB - This study aimed to evaluate the association of body fatness and sedentary status with asthma and respiratory symptoms in schoolchildren in Kaohsiung, Taiwan. A questionnaire study elicited episodes of respiratory symptoms and data on lifestyle and anthropometric parameters in 1329 5th grade schoolchildren. Results showed that 12.4% of boys and 9.5% of girls had physician-diagnosed asthma, whereas 15.1% of boys and 12.4% of girls had suspected asthma. Significantly greater proportions of boys had non-exercise-induced respiratory symptoms than girls (p < 0.05). The number of respiratory symptoms was positively correlated with TV-watching time per day and self-reported sedentary time per weekend-day in girls (p < 0.05). Underweight was positively associated with one of the seven respiratory symptoms in girls (p < 0.05). At risk of overweight was positively associated with two of the seven respiratory symptoms in boys and one of the seven respiratory symptoms in girls (all p < 0.05). The risk of having physician diagnosed asthma and suspected asthma increased 93% and 72%, respectively, in schoolchildren at risk of overweight (p < 0.05). Overweight was associated with a 78% increase in physician-diagnosed asthma (adjusted odd ratio (aOR) = 1.78, 95% CI = 1.08-2.91, p < 0.05). Higher sedentary time was significantly associated with more occurrences of one of the seven respiratory symptoms in girls (aOR = 1.05, 95% CI = 1.00-1.11, p < 0.05). Higher body mass index (BMI) was significantly correlated with longer TV-watching time per day in girls and longer self-reported sedentary time per weekday in boys (p < 0.05). In summary, schoolchildren who are at risk of overweight or overweight and/or have more sedentary time have increased risk of respiratory symptoms and asthma. Weight and sedentary statuses of schoolchildren can affect their respiratory health. PMID- 19191131 TI - Athletic trainers' experience and comfort with evaluation and management of asthma: a pilot study. AB - BACKGROUND: Approximately 10% to 50% of competitive athletes experience asthma symptoms with exercise, due to either chronic asthma or exercise-induced bronchospasm. Early recognition and management of asthma symptoms may improve athletic performance and quality of life for athletes with asthma or exercise induced bronchospasm. Athletic trainers may have frequent opportunities to identify asthma symptoms and assist athletes with management. OBJECTIVE: To survey athletic trainers about their experience and comfort with evaluation and management of asthma symptoms in athletes and identify athletic trainer characteristics associated with higher comfort levels. DESIGN AND SETTING: A 2005 cross-sectional survey of National Athletic Trainers' Association and Illinois Athletic Trainers Association members. PARTICIPANTS: A total of 304 athletic trainers. DATA COLLECTION AND ANALYSIS: Respondents completed a Web-based survey reporting years of experience, competitive level of athletes supervised, satisfaction with asthma education, experience evaluating asthma symptoms, and comfort managing asthma. RESULTS: Response rate was 13.9% (304 of 2,175). At least 23% of respondents evaluated asthma symptoms five or more times the previous year. Respondents working exclusively with junior high and/or high school athletes evaluated asthma symptoms more frequently than those working exclusively with college and/or professional athletes. Fifty-eight percent of respondents were unsatisfied with their asthma education. Only 25.3% were "very" comfortable managing asthma. Respondents with higher comfort levels evaluated asthma symptoms more frequently (p < 0.01, r = 0.18) and were more likely to be satisfied with their asthma education (p < 0.001). Over 95% of respondents endorsed more asthma education in athletic training curricula. CONCLUSIONS: Results of this pilot study indicate that athletic trainers have opportunities to help athletes manage asthma symptoms that can compromise athletic performance or limit sports participation. However, few athletic trainers are very comfortable managing asthma, and most are unsatisfied with their asthma education. Further study is needed to determine the effect of enhanced asthma education on athletic trainers' comfort and skills with asthma evaluation and management. PMID- 19191132 TI - Soluble interleukin-10 and transforming growth factor-beta in children with acute exacerbation of allergic asthma. AB - Cytokine-mediated interactions among inflammatory cells may contribute to pathogenesis of allergic asthma. To understand the role of soluble interleukin-10 (IL-10) and transforming growth factor-beta (TGF-beta) on the disease activity and regulation in asthma, changes in serum concentrations of IL-10 and TGF-beta elaborated by activated T-lymphocyte before and after prednisolone therapy with clinical improvement were determined. Circulating levels of IL-10 and TGF-beta in sera from 16 normal control subjects and in sera from 22 allergic asthmatic children with acute exacerbation and in stable condition were respectively detected by commercially available enzyme-linked immunosorbent assay kits. The mean concentrations of serum IL-10 in asthmatics with acute exacerbation (6.77 +/ 4.08 pg/mL) and during stable condition (5.14 +/- 1.17 pg/mL) were lower than that in control subjects (7.15 +/- 4.72 pg/mL). However, the difference was not statistically significant among these three study groups. The mean concentration of serum TGF-beta in stable asthmatics (40.73 +/- 15.95 pg/mL) was significantly higher than that in asthmatics with acute exacerbation (27.64 +/- 3.66 pg/mL; p < 0.05) and that in healthy control group (28.77 +/- 8.35 pg/mL; p < 0.05), while there was no statistical difference between the latter two groups. This study provides further evidence that serum TGF-beta, rather than IL-10, may play a role in regulation of disease activity and serve as an indicator for clinical control of allergic asthmatics. PMID- 19191133 TI - Determining the time to maximal bronchodilator response in asthmatic children. AB - BACKGROUND: The interval between bronchodilator administration and post bronchodilator lung function testing is critical for accurate interpretation of the bronchodilator response. The time course of this response in children is not well documented. We aimed to document the time taken to achieve maximal lung function following salbutamol inhalation. METHODS: Eighteen asthmatic children between 7 and 18 years of age with a history of bronchodilator responsiveness were recruited. Spirometry was performed before and at 0, 10, 15, 20, 40, 60, and 90 minutes after salbutamol inhalation 600 microg (Ventolin; GlaxoSmithKline) via a spacer (Volumatic; GlaxoSmithKline). RESULTS: Spirometric indices significantly increased after salbutamol inhalation (p < 0.001). The group median time to maximal response in forced expiratory volume in 1 second (FEV(1)) was 17.5 (10 60: 10th-90th centiles) minutes after salbutamol. The magnitude of group response in FEV(1) was significantly higher at 15 and 20 minutes than at 0 and 10 minutes post-salbutamol inhalation (repeat measures analysis of variance [ANOVA] on ranks; p < 0.05). CONCLUSION: We conclude that a minimal interval of 20 minutes, before re-testing spirometry, is required to document the maximal response to bronchodilators in the majority of asthmatic children. PMID- 19191134 TI - Improving asthma care for the elderly: a randomized controlled trial using a simple telephone intervention. AB - BACKGROUND: Several studies suggest that asthma is undertreated in the elderly population. OBJECTIVE: To determine if the use of a simple telephone intervention can improve asthma care in the elderly. METHODS: Fifty-two elderly subjects with asthma who required their rescue inhalers more than twice a week and had at least one emergency department or urgent care visit in the previous year were randomized to an intervention or control group. All subjects received two telephone calls over a 12-month period. The intervention group received an asthma specific questionnaire and the control group received a general health questionnaire. Medication use and health care utilization were evaluated at the beginning and end of a 12-month period. RESULTS: The study was completed by 23 control and 25 intervention subjects. Baseline data were similar in both groups. After 12 months, 72% (n = 18) of the intervention group were on an inhaled corticosteroid compared with 40% (n = 10) of the control group (p = 0.08). The intervention group had fewer emergency department visits when compared with the control group (p = 0.21). Sixty-four percent (n = 16) of the intervention group had an asthma action plan compared with 26% (n = 6) in the control group (p = 0.01). CONCLUSION: This study suggests that asthma care in the elderly can be improved using a simple telephone intervention. CLINICAL IMPLICATIONS: Clinicians need to recognize that under treatment of asthma in the elderly still exists and to use alternative methods such as a simple telephone questionnaire to improve care in this population. PMID- 19191135 TI - Impact of medical residency on knowledge of asthma. AB - OBJECTIVES: To assess resident physician knowledge of pathophysiology, diagnosis, and management of asthma and to assess the impact of an interactive curriculum on that knowledge. PARTICIPANTS: A total of 720 resident and attending physicians at 15 internal medicine residency programs. METHODS: An educational module and two multiple choice tests were developed using established methods of curriculum development and knowledge assessment, then disseminated online to 15 internal medicine residency programs. Baseline and post-intervention knowledge was analyzed according to year of training using Chi square to detect differences in group performance. RESULTS: Baseline knowledge on asthma was poor. The average baseline score on all questions was 54.2%, and was worst on questions on diagnosis of asthma (47.5% correct) and questions on management of asthma (54.8% correct). Baseline knowledge was best on questions on the pathophysiology of asthma (71.5% correct). On specific concepts, only 41.9% correctly knew which pharmacotherapeutic agents were used as controller agents, and only 43.5% were able to correctly diagnose asthma severity. Knowledge on questions on diagnosis of asthma was no better in post-graduate year (PGY) 3 residents than in PGY1 residents (p = 0.054), but PGY3 residents performed better on questions about management of asthma than did PGY1 residents (p < 0.001). Knowledge improved for all concepts and at all levels of training after completion of an interactive module on asthma guidelines (p < 0.001). CONCLUSION: Resident physician knowledge of asthma guidelines is poor and can be improved by an interactive curriculum. PMID- 19191136 TI - Matrix metalloproteinase-1 polymorphism is associated with persistent airway obstruction in asthma in the Taiwanese population. AB - BACKGROUND AND OBJECTIVE: Overexpression of matrix metalloproteinase (MMP)-1 has been demonstrated in asthma, and MMP polymorphisms are known to enhance disease susceptibility. We investigated whether MMP-1 polymorphism is associated with persistent airway obstruction in asthma in the Taiwanese population. METHODS: A total of 131 unrelated Taiwanese subjects were enrolled, age-matched, and divided as follows: (1) those who had asthma with persistent airway obstruction with forced expiratory volume in 1 second (FEV(1)) and FEV(1)/forced vital capacity (FVC) values less than 75% predicted (n = 41); (2) those with asthma without airway obstruction with FEV(1) and FEV(1)/FVC values > or = 75% predicted (n = 47); and (3) normal control subjects (n = 43). All were genotyped for the 1G/2G polymorphism of MMP-1 promoter (-1607 bp). RESULTS: 1G genotypes of MMP-1 containing at least one 1G allele were found in asthmatic patients with persistent airway obstruction (OR = 3.696, 95% CI: 1.489-9.173, p = 0.027), but not in asthmatic patients without airway obstruction (OR = 2.065, 95% CI: 0.890 4.790, p = 0.091) when compared with homozygous 2G (2G/2G). The heterozygous 1G genotype (1G/2G) was more associated with persistent airway obstruction than homozygous 2G (2G/2G) (OR: 4.727, 95% CI: 1.759-12.703, p = 0.012). The adjusted risk estimate of 1G genotypes for asthmatics with persistent airway obstruction was 4.416 (95% CI: 1.651-11.812, p = 0.003). CONCLUSION: 1G genotypes of MMP-1 polymorphism are associated with asthma with persistent airway obstruction, and the heterozygous 1G genotype (1G/2G) poses the most susceptibility to persistent airway obstruction in asthma. PMID- 19191137 TI - Antenatal steroid therapy for fetal lung maturation: is there an association with childhood asthma? AB - PURPOSE: This study was designed to test the hypothesis that fetal exposure to corticosteroids in the antenatal period is an independent risk factor for the development of asthma in childhood. METHODS: A population-based cohort study was conducted of all pregnant women who resided in Nova Scotia, Canada, and gave birth to a singleton fetus between January 1989 and December 1998 and lived to discharge. After exclusions, 79,395 infants were available for analysis. Using linked health care utilization records, incident asthma cases between 36 to 72 months of age were identified. Generalized Estimating Equations were used to estimate the odds ratio of the association between exposure to corticosteroids and asthma while controlling for confounders. RESULTS: Over the 10 years of the study corticosteroid therapy increased by threefold. Exposure to corticosteroids during pregnancy was associated with a risk of asthma in childhood: adjusted odds ratio of 1.23 (95% confidence interval: 1.06, 1.44). CONCLUSIONS: Antenatal steroid therapy appears to be an independent risk factor for the development of asthma between 36 and 72 months of age. Further research into the smallest possible steroid dose required to achieve the desired post-natal effect is needed to reduce the risk of developing childhood asthma. PMID- 19191138 TI - Are genes associated with energy metabolism important in asthma and BMI? AB - OBJECTIVE: Increased serum leptin levels have been observed in asthmatic patients. Leptin, via proliferation and activation of Th2 cells, may induce inflammation in asthma. It has also been demonstrated that leptin mRNA expression and protein levels increase in response to inflammatory stimuli. We hypothesized that polymorphisms in the leptin receptor, leptin and ghrelin genes, may affect their expression and, therefore, be responsible for altered response to increased leptin levels observed in asthma. To our knowledge, there were no studies on a potential role of LEPR, LEP, and GHRL polymorphisms in asthma. SUBJECTS AND METHODS: We analyzed 129 pediatric patients with asthma and 114 healthy children from the control group ranging from 6 to 18 years of age. The diagnosis of allergic asthma was based on clinical symptoms, the lung function test, and the positive skin prick test and/or increased immunoglobulin E (IgE) levels. Polymorphisms were genotyped by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Statistical analyses were performed with Statistica v.7.1 software (Statistica, StatSoft, Poland; available free at http://www.broad.mit.edu/mpg/haploview/index.php). Linkage disequilibrium analysis was performed with Haploview v.4.0. RESULTS: We observed a statistically significant association of the 3'UTR A/G and the -2549A/G polymorphisms of the leptin gene with asthma. No association with asthma was observed for the K109R and the Q223R polymorphisms of the LEPR gene and the Met72Leu polymorphism of the ghrelin gene. In the analysis of body mass index (BMI) stratified by genotype, we found an association of the -2549A/G LEP, but not of LEPR and GHRL polymorphisms, with higher BMI values in asthmatic patients. We found suggestive evidence for linkage between the two polymorphisms of the LEPR gene (D' = 0.84 CI: 0.71-0.92; r(2) = 0.29) in linkage disequilibrium analysis: The GG haplotype was more frequent in the control healthy group (p = 0.057). No linkage disequilibrium was detected between LEP polymorphisms. CONCLUSIONS: Polymorphisms of the leptin gene may be associated with asthma and higher BMI in asthmatic patients. Polymorphisms of the LEPR and GHRL seem unlikely to be associated with asthma or BMI in our sample. The results of haplotype analysis for the LEPR gene may suggest a protective role of the GG haplotype against asthma; however, studies on larger groups are necessary to confirm the observed trend towards association. PMID- 19191139 TI - Association of serum leptin and adiponectin with obesity in asthmatics. AB - BACKGROUND: Mounting evidence implicates obesity as a major risk factor for asthma. Leptin and adiponectin produced by fat tissues play a critical role in the regulation of body weight and allergic inflammation. OBJECTIVE: The aim of this study was to evaluate the effects of leptin and adiponectin on development of asthma. METHODS: We measured the leptin and adiponectin in serum from patients with asthma (n = 60) and normal control subjects (n = 30) by enzyme-linked immunosorbent assay. RESULTS: Logarithmic leptin and adiponectin concentration was not different between asthmatics and control subjects. Although the logarithmic adiponectin level was not different by gender in asthmatics, the logarithmic leptin concentration was significantly higher in females than in male asthmatics (2.41 +/- 0.05 ng/mL vs. 2.01 +/- 0.05 ng/mL, p = 0.001). The leptin/adiponectin ratio was also significantly higher in females than in male asthmatics. The leptin/adiponectin ratio was correlated with body mass index (r = 0.210, p = 0.05) in asthmatics. CONCLUSION: Our results suggest that serum leptin and adiponectin may be associated with gender and obesity regardless of development of asthma. PMID- 19191140 TI - Beta-(1,3)-glucan levels and its determinants in New Zealand bedrooms. AB - Beta-(1,3)-glucan is a pro-inflammatory component of the fungal cell wall, indoor levels of which have only been considered in a few studies. This study assessed levels of beta-(1,3)-glucan in 35 domestic bedrooms. beta-(1,3)-glucan levels were estimated using a modified Limulus amoebocyte lysate kinetic assay. beta (1,3)-glucan geometric mean levels (95% confidence interval) were 163.9 microg/g (129.5-209.3) from bedroom floors; 76,6 microg/g (61.4-94.0) from mattresses; 132.1 microg/g (68.9-207.9) from duvets; and 110.0 microg/g (82.2-146.4) from pillows. Synthetic bedding and older mattresses contained higher beta-(1,3) glucan levels. Synthetic bedding contains higher levels of beta-(1,3)-glucan than feather bedding, which may be of importance to asthmatics. PMID- 19191141 TI - Asthma severity categorization using a claims-based algorithm or pulmonary function testing. AB - OBJECTIVES: This study was performed to determine whether pulmonary function test results would appreciably alter asthma severity categorization determined by an algorithm using information readily available in administrative databases. METHODS: Patients 6 to 64 years of age with asthma diagnosed from 1999-2005, who had at least one pulmonary function test, were identified from a claims database of a medical group practice located in central Massachusetts. Asthma severity for these patients was categorized using information available in an administrative database (claims-based algorithm) and by percent predicted forced expiratory volume in 1 second (FEV(1)) or peak expiratory flow (PEF) abstracted from medical charts (pulmonary function test method). Gamma rank correlation index was used to measure the association between the two severity categorization methods. Total and asthma-related healthcare costs for each severity category were compared between the two different approaches. RESULTS: There was a significant ordinal association between severity categorization with the two classification approaches (p = 0.0002). The pulmonary function test method resulted in more frequent mild categorizations and less frequent moderate and severe categorizations than the claims-based algorithm. In only 10.9% of patients did the pulmonary function test method result in a more severe asthma category than the claims-based algorithm. Patients with more severe asthma, determined by both methods, had higher total and asthma-related health care costs. Total and asthma related health care costs were similar for each asthma severity categorization for the two classification approaches, except for asthma-related costs in the moderate severity categories. CONCLUSION: The claims-based algorithm generally categorized patients as having more severe asthma than the approach using pulmonary function test results. Pulmonary function test results would have appreciably changed asthma severity categorization in only a small percent of patients. These findings add further support to the use of administrative database analyses for the evaluation of asthma care in large populations. PMID- 19191142 TI - Incremental direct expenditure of treating asthma in the United States. AB - OBJECTIVE: There is a wide range in the estimates of cost of asthma that are available in the literature. Given the growing prevalence of asthma and its associated healthcare resource use in the United States (U.S.), it is important to obtain current and precise cost estimates attributable to asthma treatment. The objectives of this study were to estimate the incremental direct expenditures associated with asthma in the U.S. METHODS: Retrospective analysis was conducted using the 2004 Medical Expenditure Panel Survey (MEPS) data that are representative of the civilian non-institutionalized population of the U.S. Asthma respondents were identified as those with International Classification of Diseases-9-Clinical Modification (ICD-9-CM) diagnosis codes for asthma in 2004 or those who had a self-report of having asthma in 2004. Incremental total expenditures and expenditures for various categories of resource use including physician office visits, emergency room visits, outpatient visits, inpatient visits, medications, and other medical visits associated with asthma were estimated separately in children (age < 18 years) and in adults (age > or = 18 years) using generalized linear regression models. The models were adjusted for covariates including age, gender, race, ethnicity, education, marital status (for age group > or = 18 years), geographic region, insurance status, and comorbidities. RESULTS: The prevalence of asthma among children and adults in 2004 was estimated at 8.7% (6.4 million persons) and 6.72% (14.8 million persons), respectively. The annual adjusted mean incremental total expenditure associated with asthma was $1,004.6 (SE: $326.1; p = 0.002) per person among children and was $2,077.5 (SE: $544.5; p < 0.0001) per person among adults, after adjusting for covariates. Prescription medications and physician office visits were the major drivers of total expenditures and constituted approximately 38% and 49% of the total incremental expenditures for asthma in children and adults, respectively. Inpatient visit expenditures were high in both age groups but were not significantly different from zero. CONCLUSION: Given the prevalence of asthma among U.S. children and adults and its associated incremental expenditures, the annual direct medical expenditure attributable to asthma treatment is estimated at approximately $37.2 billion in 2007 U.S. dollars representing a significant portion of healthcare resource use in the U.S. PMID- 19191143 TI - Iron and ferritin concentrations in exhaled breath condensate of children with asthma. AB - Maintenance of iron homeostasis is of utmost importance for the respiratory system physiology and pathophysiology. Local iron deficiency or accumulation may result in particular respiratory function impairment. The aim of the present study was to find out whether iron and ferritin could be determined in exhaled breath condensate (EBC) of healthy children and children with asthma. Oxidative stress was verified by determination of EBC superoxide dismutase (SOD) activity, and the airway inflammatory process by determination of exhaled nitric oxide (F(E)NO). EBC was collected from 39 children (22 healthy children as a control group and 17 asthmatics) using an EcoScreen condenser. Iron, ferritin, and SOD were determined on optimization and validation for low concentrations. In comparison with a control group, asthma patients had a statistically significantly lower iron concentration (p = 0.0001) and higher SOD catalytic activity (p = 0.0160), with no significant difference in ferritin levels (p = 0.5252), although percentile values indicated elevated ferritin concentration in about half of asthma patients. F(E)NO values were significantly higher in the asthma group (p = 0.0047). This preliminary study demonstrated the possibility of determining iron and ferritin concentrations and SOD activity in EBC, and a significant difference in EBC iron and SOD between asthma patients and healthy children. PMID- 19191144 TI - Prevalence of asthma and allergic rhinitis among school children of Karachi, Pakistan, 2007. AB - In recent times, the incidence of allergic diseases, particularly bronchial asthma, has been increasing worldwide. However, there appears to be no published data on the prevalence of allergic diseases among school children (3 to 16 years of age) in Karachi, Pakistan, with only limited data available among few age groups under one ISAAC study. The objective of this project was to investigate the prevalence of allergic diseases among school children (3 to 16 years of age) in the city of Karachi. The questionnaire that was used for data collection had previously been used for a similar study in neighboring Saudi Arabia and the U.A.E. In 2007, a total of 3,000 surveys were distributed in various schools of Karachi, of which 2,325 completed surveys were obtained. SPSS was used to perform statistical analysis on the collected data. Survey results showed that the frequency of diagnosed (previously seen by physicians) cases of asthma stood at 15.8%, while the frequency of allergic rhinitis was found to be 28.50% among these children. Other parameters that were analyzed included dry cough (20.1%), wheezing (11.7%), breathlessness (15.40%), and eczema (21.8%). Furthermore, smoking by family members was found to be associated with asthma (p value less than 0.05), allergic rhinitis (p value less than 0.05), breathlessness (p value less than 0.05), dry cough (p value 0.002), and wheezing (p value less than 0.05). This study reveals that there is a significant number of school children in the metropolitan city of Karachi who have various allergic symptoms. It also sheds light on the fact that exposure to indoor environmental factors as well as family atopy can play a key role in increasing the chances of an individual to experience asthma and other allergy symptoms. PMID- 19191145 TI - A population-based clinical study of allergic and non-allergic asthma. AB - BACKGROUND: The aim of this study was to describe differences between allergic and non-allergic asthma in a large community-based sample of Danish adolescents and adults. METHODS: A total of 1,186 subjects, 14 to 44 years of age, who in a screening questionnaire had reported a history of airway symptoms suggestive of asthma and/or allergy, or who were taking any medication for these conditions were clinically examined. All participants were interviewed about respiratory symptoms, and furthermore skin test reactivity, lung function, and airway responsiveness were measured. RESULTS: A total of 489 individuals had clinical asthma of whom 61% had allergic asthma, whereas 39% had non-allergic asthma. Subjects with non-allergic asthma were more likely to be females, OR = 2.24 (1.32 3.72), p = 0.003, and to have cough as the predominant symptom, OR = 1.96, (1.19 3.23), p = 0.008, but were less likely to have AHR, OR = 0.40, (0.24-0.66), p < 0.001, food allergy, OR = 0.28, (0.11-0.73), p = 0.009, and symptoms of rhinitis, OR = 0.08 (0.05-0.14) compared with subjects with allergic asthma. Subjects with non-allergic asthma had had persistent symptoms within the last 4 weeks more often than subjects with allergic asthma (68% vs. 53%), p = 0.001. CONCLUSIONS: Non-allergic asthma accounts for two in every five cases of asthma in adults and constitutes symptomatically, and in terms of lung function, a more severe form of disease than allergic asthma. PMID- 19191146 TI - Eosinophilic pneumonia caused by Aspergillus niger: is oral cleansing with amphotericin B efficacious in preventing relapse of allergic pneumonitis? AB - Eosinophilic pneumonia was confirmed by bronchoalveolar lavage fluid examination and transbronchial lung biopsy. Aspergillus niger was cultured from the patient's pharyngeal swab and bronchoalveolar lavage fluid. Inhalation bronchoprovocation test with A. niger antigen was positive. Although the patient's condition improved promptly with 10 mg/day prednisolone administration, dry cough recurred approximately 2 months after completion of this therapy. Severe coughing disappeared on oral cleansing with 300 mg/day amphotericin B, and he recovered completely on 100 mg/day amphotericin B administration. Oral cleansing with amphotericin B may be efficacious in preventing relapses of eosinophilic pneumonia caused by allergic reaction to fungal antigen. PMID- 19191148 TI - Reduction of the total IgE Level by omalizumab in children and adolescents. PMID- 19191150 TI - Gentleman surgeon. AB - The gentleman surgeon is the individual who demonstrates that he/she cares, respects, and helps all of those involved in the profession, from the practicing surgeon, fellow resident, student, operating room personnel, and floor nurse to patients in treatment. Surgeons who care, help, and respect others in their daily activities are worthy to be considered a gentleman surgeon. From the example of surgical pioneers who practiced the principles of the gentleman surgeon to the surgeon of tomorrow, we review those important characteristics associated with the humanistic approach to becoming a gentleman surgeon, that is the surgeon who cares, respects, and helps. PMID- 19191151 TI - The strange little animals of Antony van Leeuwenhoek surgical revolution. AB - Antony van Leeuwenhoek (1632-1723) created a surgical revolution by demonstrating, through a series of extraordinary discoveries, the presence of "strange little animals" under the microscope. His outstanding advances were directly related to his ability to grind better glasses, which enhanced magnification many times over previously manufactured glasses. His meticulous and dedicated observational skills were unmatched by anyone dealing with magnification at the time. The surgical revolution did not occur during Leeuwenhoek's time but more than a century later when the value of his findings was evidently recognized. Today Leeuwenhoek is considered the father of microscopy as well. It is particularly enlightening that for not being a scientist himself, he demonstrated all the good virtues of method and technique for which professional scientists are admired. PMID- 19191152 TI - Heat shock treatment prior to myocardial infarction results in reduced ventricular remodeling. AB - BACKGROUND: Heat shock (HS) treatment has been suggested to confer myocardial protection following ischemia. However, the effects of HS on left ventricular (LV) remodeling weeks after infarction have yet to be described. METHODS: Myocardial infarction (MI) was created by coronary ligation in Lewis rats. Two experimental groups of animals were created: HS+MI group (n = 13) and MI group (n = 13). HS treatment consisted of an elevation in core temperature to 42 degrees C for 15 min, 24 hr prior to MI. LV remodeling was assessed by transthoracic echocardiography (day 0, 1, 7, and 28) and by morphometric histology (day 28). RESULTS: There was no significant difference in infarct size (TTC stain 24 hr) between HS+MI and MI groups. Using transthoracic echo there was a significant preservation of LV ejection fraction and fractional shortening in the HS+MI group as compared to MI group (7 and 28 days). Similar trends were seen by histology at 28 days but failed to reach significance. HSP27 expression by myocardial cells was shown to remain up-regulated (at 28 days) in both groups at the edges of the infarct area as compared to control myocardium. CONCLUSIONS: Our findings suggest that HS treatment prior to MI can result in a significant decrease in LV remodeling independent of a reduction in infarct size. PMID- 19191153 TI - Femoral venous catheterization is a major risk factor for central venous catheter related bloodstream infection. AB - BACKGROUND: Bloodstream infection is a major complication associated with central venous catheters (CVCs). However, there have been few studies of the risk factors for catheter-related bloodstream infection in patients who undergo colorectal surgery (CRS). PURPOSE: To disclose the risk factors for catheter-related bloodstream infection in CRS. METHODS: Catheter-related bloodstream infection was evaluated retrospectively from a database of patients who had undergone CRS. RESULTS: Three hundred-fifty patients received 423 CVCs for a total of 7,760 catheter days. Thirty-nine cases of catheter-related bloodstream infection (5.03 per 1,000 catheter days) were diagnosed. There were no significant differences in background between patients with or without catheter-related bloodstream infection, with the exception of the term of catheter insertion (24.6 +/- 7.0 days vs.17.7 +/- 0.6 days, P = .0151). However, univariate analysis using factors of sex, age, insertion difficulty, length of the inserted catheter, term of catheter insertion, administration of chemotherapy, administration of total parenteral nutrition (TPN), kind of disinfectant, degree of surgical insult, and type of catheter revealed that use of a femoral venous catheter was an independent risk factor for catheter-related bloodstream infection (odds ratio [OR] = 3.175; 95% confidence interval [CI], 1.103-9.139; P = .0322). CONCLUSIONS: Use of femoral venous catheters is a major risk factor for catheter-related bloodstream infection in CRS. PMID- 19191154 TI - Mild hypothermia improves survival during hemorrhagic shock without affecting bacterial translocation. AB - BACKGROUND: Accidental hypothermia in patients with hemorrhagic shock (HS) is associated with increased mortality. However, experimental mild and moderate hypothermia has beneficial effects. The mechanisms for beneficial effects of hypothermia have not been completely understood. Therefore, the aim of this study was to investigate the effect of hypothermia on survival, bacterial translocation (BT), and remote pulmonary injury in a controlled HS model in rats. METHODS: HS was achieved by blood withdrawal through femoral vein. Rats in the normothermia group (group I) were maintained at 37 degrees C. Mild hypothermia group (group II) was observed at 32 degrees C that was spontaneously induced by exposure to ambient temperature. Moderate hypothermia of 28 degrees C was actively induced by external cooling in group III for 90 min. Survival and neurological deficit scores (NDS) were recorded at 24th hr. Mesenteric lymph nodes, liver and spleen samples were collected. Myeloperoxidase (MPO) and malondialdehyde (MDA) levels were measured in lung tissue. RESULTS: Blood pressure significantly increased in hypothermia groups. Mild hypothermia significantly increased survival. No difference was found in BT rates in groups. Hypothermia was found to significantly decrease the NDS points in group III, compared to group I. There was no difference in lung tissue MPO levels among groups. Lung tissue MDA levels increased significantly in groups II and III. CONCLUSIONS: Mild hypothermia improved blood pressure, survival, and neurological outcome with a possible detrimental effect on pulmonary ROS production during HS in rats. These effects of hypothermia are not associated with BT. PMID- 19191155 TI - Effect of endoplasmic reticulum stress in VEGF-induced neuroprotection. AB - Researchers suggest that endoplasmic reticulum (ER) stress cause apoptosis after ischemia. Caspase-12 has been localized to the ER and is a signal for apoptosis. We sought to clarify the role of caspase-12 in the vascular endothelial growth factor (VEGF) induced neuroprotective effect. Transient focal cerebral ischemia was produced by occluding left middle cerebral artery in rabbit. The expressions of caspase-12 and caspase-3 were detected by immunohistochemistry. Neuronal apoptosis was detected by TUNEL staining. We confirmed that the number of apoptotic cells and the expressions of caspase-12 and caspase-3 significantly increased during reperfusion. VEGF inhibited the cell apoptosis and the expressions of caspase-12 and caspase-3. These results suggest that VEGF may protect neurons from apoptosis by inhibiting ER stress pathway. PMID- 19191156 TI - Focused in vivo genetic analysis of implanted engineered myofascial constructs. AB - Successfully engineering functional muscle tissue either in vitro or in vivo to treat muscle defects rather than using the host muscle transfer would be revolutionary. Tissue engineering is on the cutting edge of biomedical research, bridging a gap between the clinic and the bench top. A new focus on skeletal muscle tissue engineering has led investigators to explore the application of satellite cells (autologous muscle precursor cells) as a vehicle for engineering tissues either in vitro or in vivo. However, few skeletal muscle tissue engineering studies have reported on successful generation of living tissue substitutes for functional skeletal muscle replacement. Our model system combines a novel aligned collagen tube and autologous skeletal muscle satellite cells to create an engineered tissue repair for a surgically created ventral hernia as previously reported [SA Fann, L Terracio, W Yan, et al., A model of tissue engineered ventral hernia repair, J Invest Surg. 2006;19(3):193-205]. Several key features we specifically observe are the significant persistence of transplanted skeletal muscle cell mass within the engineered repair, the integration of new tissue with adjacent native muscle, and the presence of significant neovascularization. In this study, we report on our experience investigating the genetic signals important to the integration of neoskeletal muscle tissue. The knowledge gained from our model system applies to the repair of severely injured extremities, maxillofacial reconstructions, and restorative procedures following tumor excision in other areas of the body. PMID- 19191157 TI - Nitric oxide mechanism of protection in ischemia and reperfusion injury. AB - In 1992 nitric oxide (NO) was declared molecule of the year by Science magazine, and ever since research on this molecule continues to increase. Following this award, NO was shown to be a mediator/protector of ischemia and reperfusion injury in many organs, such as the heart, liver, lungs, and kidneys. Controversy has existed concerning the actual protective effects of NO. However, literature from the past 15 years seems to reinforce the consensus that NO is indeed protective. Some of the protective actions of NO in ischemia and reperfusion are due to its potential as an antioxidant and anti-inflammatory agent, along with its beneficial effects on cell signaling and inhibition of nuclear proteins, such as NF-kappa B and AP-1. New therapeutic potentials for this drug are also continuously emerging. Exogenous NO and endogenous NO may both play protective roles during ischemia and reperfusion injury. Sodium nitroprusside and nitroglycerin have been used clinically with much success; though only recently have they been tested and proven effective in attenuating some of the injuries associated with ischemia and reperfusion. NO inhalation has, in the past, mostly been used for its pulmonary effects, but has also recently been shown to be protective in other organs. The potential of NO in the treatment of ischemic disease is only just being realized. Elucidation of the mechanism by which NO exerts its protective effects needs further investigation. Therefore, this paper will focus on the mechanistic actions of NO in ischemia and reperfusion injury, along with the compound's potential therapeutic benefits. PMID- 19191158 TI - A method for the consistent creation and quantitative testing of postoperative pelvic adhesions in a porcine model. AB - INTRODUCTION: The assessment of methods for the prevention of postoperative pelvic adhesions is hampered by the lack of a quantifiable adhesion measurement technique. Currently available methods for adhesion model assessment rely on qualitative grading scales; a quantitative method would have many attendant benefits, including standardized reporting. METHODS: A technique was developed to generate consistent and significant adhesions in a swine model that are suitable for quantitative assessment using a Material Testing System (MTS) machine platform. In this method, the uterine horns are cannulated and then attached to the pelvic sidewall using loose silk sutures. The underlying sidewall and adjacent uterine serosa are injured with electrocautery. Following a two-week survival the entire complex of uterine horn and sidewall are excised en bloc and prepared for MTS testing. RESULTS: Extrication of the adhesion, as measured by the MTS platform, generates a quantitative assessment of adhesion strength that can be scaled and analyzed to produce several continuous variable descriptions. DISCUSSION: This technique represents a novel quantitative method for adhesion assessment in an animal model. This quantitative technique may then be applied to accurately assess a range of adhesion prevention techniques, producing results which can be standardized for comparison. PMID- 19191159 TI - Two-staged procedure of portal ligation and hepatectomy monitored by ICG clearance. AB - BACKGROUND: The aim of the study was to predict and estimate the optimal time of resection following portal vein ligation (PVL) with the aid of three dimensional (3D) computed tomography (CT) volumetry and indocyanine green (ICG) clearance in healthy liver. Following right PVL, we induced hypertrophy of the left liver lobe prior to an extended right hepatectomy. METHODS: Fourteen patients with right lobe liver metastases from colorectal cancer underwent right PVL. Cutoff points for the ICG clearance test were: R15 < 14% or PDR > 15%/min. The criteria for resection also required a remnant liver volume of 25% of the whole liver volume. The latter was assessed by CT scan prior to the procedures. Liver function was measured with routine biochemical tests and ICG clearance. Postoperatively, repeated ICG clearance and 3D CT volumetry tests were used to estimate the liver's regeneration. Liver resections were performed as a second stage. RESULTS: After portal ligation, ICG clearance increased significantly in some patients, while in others, the ICG clearance remained unchanged with borderline low or normal clearance values. Between the two operations, patients with high clearance had less complications and a better regeneration rate of the left lobe with a shorter waiting period in contrast to the "low ICG group." CONCLUSIONS: ICG clearance has a significant prognostic value. Patients with an apparently inoperable right lobe liver tumor can be successfully treated using a two-staged procedure of portal vain ligation followed by hepatectomy. The 3D CT volumetry and ICG clearance test are essential monitoring tools in these liver resections. PMID- 19191160 TI - Monitoring of experimental rat lung transplants by high-resolution flat-panel volumetric computer tomography (fpVCT). AB - OBJECTIVE: Noninvasive assessment of experimental lung transplants with high resolution would be favorable to exclude technical failure and to follow up graft outcome in the living animal. Here we describe a flat-panel Volumetric Computed Tomography (fpVCT) technique using a prototype scanner. METHODS: Lung transplantation was performed in allogeneic as well as in corresponding syngeneic rat strain combinations. At different time points post-transplantation, fpVCT was performed. RESULTS: Lung transplants can be visualized in the living rat with high-spatial resolution. FpVCT allows a detailed analysis of the lung and the bronchi. Infiltrates developing during rejection episodes can be diagnosed and follow-up studies can easily be performed. CONCLUSIONS: With fpVCT it is possible to control the technical success of the surgical procedure. Graft rejection can be visualized individually in the living animal noninvasively, which is highly advantageous for studying the pathogenesis of chronic rejection or to monitor new therapies. PMID- 19191161 TI - Principles of anesthesia monitoring--respiration. PMID- 19191163 TI - Measurements of airborne methylene diphenyl diisocyanate (MDI) concentration in the U.S. workplace. AB - This article summarizes a large body of industry air sampling data (8134 samples) in which airborne MDI concentrations were measured in a wide variety of manufacturing processes that use either polymeric MDI (PMDI) or monomeric (pure) MDI. Data were collected during the period 1984 through 1999. A total of 606 surveys were conducted for 251 companies at 317 facilities. The database includes 3583 personal (breathing zone) samples and 4551 area samples. Data demonstrate that workplace airborne MDI concentrations are extremely low in a majority of the manufacturing operations. Most (74.6%) of the airborne MDI concentrations measured in the personal samples were nondetectable, i.e., below the limits of quantification (LOQs). A variety of validated industrial hygiene sampling/analytical methods were used for data collection; most are modifications of OSHA Method 47. The LOQs for these methods ranged from 0.1-0.5 microg/sample. The very low vapor pressures of both monomeric MDI and PMDI largely explain the low airborne concentrations found in most operations. However, processes or applications in which the chemical is sprayed or heated may result in higher airborne concentrations and higher exposure potentials if appropriate control measures are not implemented. Data presented in this article will be a useful reference for employers in helping them to manage their health and safety program as it relates to respiratory protection during MDI/PMDI applications. PMID- 19191164 TI - Effectiveness of common shelter-in-place techniques in reducing ammonia exposure following accidental release. AB - Shelter-in-place strategies such as remaining indoors; breathing through a damp cloth; sealing cracks in windows and doors using towels, duct tape, or plastic sheeting; and running a shower are often recommended by emergency response officials to protect against accidental or intentional release of hazardous airborne chemicals and biologicals. Similar recommendations have been made to and used by community members exposed to anhydrous ammonia after catastrophic release of ammonia gas due to a derailment or other accidents. Such incidents have resulted in fatalities and serious injury to exposed individuals; however, other individuals within the same area have escaped injury and, in many cases, sustained no injuries as a result of sheltering-in-place. Although there are some studies that have evaluated the effectiveness of remaining in the home or breathing through a damp cloth to reduce exposure to various agents, there have been no studies that directly address the efficacy of running the shower in reducing exposure to ammonia gas. The present study was designed to simulate sheltering-in-place inside a typical bathroom with the shower running. The effectiveness of breathing through a damp cloth was also evaluated using a CPR mannequin placed inside a chamber built to represent a typical household bathroom. Ammonia gas at 300 or 1000 ppm was added to the chamber until the concentration peaked and stabilized, then the shower was turned on and the ammonia gas concentration was continuously monitored. In the mannequin studies, using a damp cloth reduced exposure to ammonia gas by 2- to 18-fold. Turning on the shower was even more effective at reducing ammonia levels. After 27 min, the ammonia concentration in the chamber was reduced to 2% of the initial concentration, even though gas was being continuously added to the chamber. These results indicate that use of shelter-in-place strategies substantially reduces ammonia exposure and that by combining shelter-in-place strategies, inhalation of ammonia gas can be reduced 100-fold even during prolonged exposure periods. PMID- 19191165 TI - Causal attribution and emotion in the days following competition. AB - We examined the extent to which attributions are consistent in the days following competition and how attributions made immediately after competition may influence the temporal patterning of emotions experienced in response to competition. A sample of 60 adult female golfers completed measures of performance satisfaction, causal attribution, and emotion immediately after competition, 5 h after competition, and 2 days after competition. Repeated-measures multivariate analysis of covariance indicated that attributions did not change over this period. Emotions showed a significant decrease in intensity over the 5-h post competition period. Regression analyses indicated that changes in anger and dejection were more likely in the case of less successful performances. For anger, attributions moderated this level of change. Golfers experienced anger for a longer period when they identified the cause of poor performance as stable rather than unstable. Thus, in the present sample although attributions did not change over 2 days, the longevity of anger depended on the attributions made immediately after competition. PMID- 19191166 TI - Talent identification and deliberate programming in skeleton: ice novice to Winter Olympian in 14 months. AB - The aims of this study were to talent transfer, rapidly develop, and qualify an Australian female athlete in the skeleton event at the 2006 Torino Winter Olympic Games and quantify the volume of skeleton-specific training and competition that would enable this to be achieved. Initially, 26 athletes were recruited through a talent identification programme based on their 30-m sprint time. After attending a selection camp, 10 athletes were invited to undertake an intensified skeleton training programme. Four of these athletes were then selected to compete for Australia on the World Cup circuit. All completed runs and simulated push starts were documented over a 14-month period. The athlete who eventually represented Australia at the Torino Winter Olympic Games did so following approximately 300 start simulations and about 220 training/competition runs over a period of 14 months. Using a deliberate programming model, these findings provide a guide to the minimum exposure required for a novice skeleton athlete to reach Olympic representative standard following intensified sport-specific training. The findings of this study are discussed in the context of the deliberate practice theory and offer the term "deliberate programming" as an alternative way of incorporating all aspects of expert development. PMID- 19191167 TI - Performance of Chromogenic Candida agar and CHROMagar Candida in recovery and presumptive identification of monofungal and polyfungal vaginal isolates. AB - Chromogenic Candida agar (OCCA) is a novel medium facilitating isolation and identification of Candida albicans, C. tropicalis, and C. krusei, as well as indicating polyfungal population in clinical samples. We compare the performance of OCCA, to CHROMagar Candida (CAC) and Sabouraud chloramphenicol agar (SCA). Vaginal swab samples from 392 women were simultaneously inoculated onto three study media. A total of 161 (41.1%) were found to be positive for fungi of which 140 (87%) were monofungal, and 21 (13%) polyfungal. One-hundred and fifty-seven samples (97.5%) were positive on CAC, 156 (96.9%) on OCCA, 148 (91.9%) on SCA and 144 (89.4%) samples were positive on all three media. The yeasts were identified by conventional methods including germ tube test, microscopic morphology on cornmeal-Tween 80 agar, and the commercial API 20C AUX. The 182 isolates were C. albicans (n = 104), C. glabrata (n = 51), C. krusei (n = 7), C. tropicalis (n = 5), C. famata (n = 3), C. kefyr (n = 3), C. zeylanoides (n = 3), C. colliculosa (n = 2), and other species of Candida (n = 4). Among the 21 polyfungal populations, 20 (95.2%) were detected in OCCA, 14 (66.7%) in CAC, and 13 (61.9%) in CAC and OCCA (P <0.05). Most polyfungal populations (47.6%) yielded C. albicans + C. glabrata. The efficiency of both chromogenic media for C. albicans was >or=92.9% at 72 h. OCCA is more efficient and reliable for rapidly identifying C. albicans and polyfungal populations than CAC. However, CAC is more efficient for identifying C. krusei and C. tropicalis. A chromogenic agar with a higher isolation rate of yeasts and better detection of polyfungal populations than SCA, is suggested as a medium of first choice when available. PMID- 19191169 TI - Scedosporium apiospermum skeletal infection in an immunocompetent child. AB - Abstract This is a case of Scedosporium apiospermum skeletal infection in a 10 year-old immunocompetent girl whose chief complaint was left knee swelling and pain. The child had a history of a bicycle accident two months before with a resultant deep penetrating trauma. Systematic administration of broad-spectrum antibiotics for 10 days was used, with no clinical improvement. Magnetic Resonance Imaging and arthrotomy of the affected joint revealed findings suggestive of osteomyelitis. Empirical intravenous antimicrobial therapy was instituted for a total of two months but one month after completion of antibacterial therapy the child returned to the hospital because of persistent knee swelling and pain. Following a new arthrotomy, Scedosporium apiospermum was isolated. The patient was cured with intravenous administration of voriconazole without any side effects and has no evidence of relapse after four years of follow-up. PMID- 19191171 TI - Cell adhesion under flow. AB - Cell adhesion under flow is a central function of the microcirculation during inflammation, hemostasis, and immune regulation. This special issue of Microcirculation explores the common and distinct mechanisms that myeloid cells, lymphocytes, platelets, and sickle erythrocytes use to adhere to microvascular endothelium and the underlying basement membrane structures. A common theme in these processes is the need for rapid integrin activation, often initiated by binding of ligands to their cognate G protein-coupled receptors, followed by adhesion strengthening associated with integrin redistribution and outside-in signaling. These elements have been identified for all cells tested except sickle erythrocytes. PMID- 19191172 TI - Palmitoyl acyltransferases, their substrates, and novel assays to connect them (Review). AB - Thio-palmitoylation is the post-translational addition of the 16-carbon fatty acid, palmitate, to the thiol side chain of cysteine residues by a labile thioester bond. Palmitoylation increases the lipophilicity of a protein resulting in dramatic changes in its subcellular distribution such as moving from the endoplasmic reticulum to the plasma membrane or in subtle changes like an increased affinity for cholesterol-rich lipid rafts in membranes. Palmitoylation is also dynamic, making it unique among post-translational protein lipid modifications. Discovering the molecular identity of palmitoyl acyltransferases (PATs) was a watershed event that dramatically accelerated the pace of discovery in the field. Likewise, there has been increased interest in palmitoylation partly because many of the genes encoding PATs have been linked to cancer and other diseases. Now, with a greater understanding of how palmitate is enzymatically attached to proteins, some of the most interesting questions include: What are the substrates of each PAT?; how does a PAT recognize and palmitoylate a substrate?; how are PATs regulated?; and, how is depalmitoylation regulated? The answers to these questions are beginning to unfold due to the recent development of novel assays as well as the expansion and refinement of existing assays. Our ability to understand palmitoylation and its importance to human health and disease is only as good as the methods we use to test our hypotheses. The continued development of methods with increased sensitivity and selectivity is critical to this venture. PMID- 19191173 TI - Protein S-acylation in plants (Review). AB - Membrane resident proteins are a common feature of biology yet many of these proteins are not integral to the membrane. These peripheral membrane proteins are often bound to the membrane by the addition of fatty acyl chains to the protein. This modification, known as S-acylation or palmitoylation, promotes very strong membrane association but is also reversible allowing for a high degree of control over membrane association. Many S-acylated proteins are resident in sterol, sphingolipid and saturated-lipid enriched microdomains indicating an important role for S-acylation in protein partitioning within membranes. This review summarises the current knowledge of S-acylation in plants. S-acylated proteins play a wide variety of roles in plants and affect Ca(2+) signalling, K(+) movement, stress signalling, small and heterotrimeric G-protein membrane association and partitioning, tubulin function as well as pathogenesis. Although the study of S-acylation is in its infancy in plants this review illustrates that S-acylation is extremely important for plant function and that there are many unexplored aspects of S-acylation in plants. A full summary of the techniques and methods available to study S-acylation in plants is also presented. PMID- 19191170 TI - Platelet adhesion under flow. AB - Platelet-adhesive mechanisms play a well-defined role in hemostasis and thrombosis, but evidence continues to emerge for a relevant contribution to other pathophysiological processes, including inflammation, immune-mediated responses to microbial and viral pathogens, and cancer metastasis. Hemostasis and thrombosis are related aspects of the response to vascular injury, but the former protects from bleeding after trauma, while the latter is a disease mechanism. In either situation, adhesive interactions mediated by specific membrane receptors support the initial attachment of single platelets to cellular and extracellular matrix constituents of the vessel wall and tissues. In the subsequent steps of thrombus growth and stabilization, adhesive interactions mediate platelet-to platelet cohesion (i.e., aggregation) and anchoring to the fibrin clot. A key functional aspect of platelets is their ability to circulate in a quiescent state surveying the integrity of the inner vascular surface, coupled to a prompt reaction wherever alterations are detected. In many respects, therefore, platelet adhesion to vascular wall structures, to one another, or to other blood cells are facets of the same fundamental biological process. The adaptation of platelet adhesive functions to the effects of blood flow is the main focus of this review. PMID- 19191174 TI - The continuing challenge of vitamin A deficiency. PMID- 19191175 TI - Incidence of blinding vitamin A deficiency in North West Frontier Province and its adjoining Federally Administered Tribal Areas, Pakistan. AB - PURPOSE: To identify children aged 0-72 months with blinding xerophthalmia in the North West Frontier Province (NWFP) and its adjoining Federally Administered Tribal Areas (FATA) in Pakistan, using a new surveillance system, and to describe socio-economic and other characteristics of reported cases. METHODS: All ophthalmologists in district headquarter hospitals in NWFP and agency headquarter hospitals in FATA were requested to participate in a surveillance programme for the detection of blinding xerophthalmia (corneal xerosis, corneal ulcers, keratomalacia and corneal scars related to vitamin A deficiency) for children aged 0-72 months. The surveillance included reporting of each case of blinding xerophthalmia presenting to the participating eye units over a period of 12 months and recording their socio-economic and other related characteristics. RESULTS: Seventy six cases of blinding xerophthalmia were reported and there was almost no gender difference. Children came from 19 of 31 districts and agencies in NWFP and FATA with a working ophthalmologist. The commonest age at presentation was 25-48 months and almost half of them belonged to households with a monthly income of < Pak Rs. 1200 (US $ < 20). Precipitating systemic illnesses were diarrhea and fever with cough in 63.2% and 56.6% cases, respectively. Almost a third of children aged > 9 months had not been immunized against measles. A quarter of all children had not been adequately breast-fed, and 7 children (9%) had never been breastfed. Majority of the cases presented during summer and autumn months. CONCLUSIONS: Our surveillance data provide evidence that blinding xerophthalmia does occur in children in NWFP and its adjoining FATA. The surveillance system identified high risk age groups, systemic illnesses, seasons and community groups. The detection system detects minimal numbers of cases and it is likely that the problem is even more severe. PMID- 19191176 TI - Prevalence and determinants of xerophthalmia in preschool children in urban slums, Pune, India--a preliminary assessment. AB - PURPOSE: International and national programs to control vitamin A deficiency disorders (VADD) among children in developing countries are reducing the magnitude but VADD still occurs in deprived populations. The purpose of the study was to estimate the prevalence of and identify risk factors for xerophthalmia in children aged 6-71 months living in slums in Pune, India in 2003. METHODS: Children were enrolled into a cross sectional study from randomly selected Anganwadis (kindergartens) in selected slums using proportional to size sampling. Parents/caregivers were interviewed by social workers about night blindness, risk factors for VADD at individual and household levels, intake of vitamin A rich foods, and history of vitamin A supplementation. Children were examined for signs of xerophthalmia by an ophthalmologist. Risk factors for xerophthalmia were explored using univariate and multivariate regression analysis. RESULTS: 1,589 children were examined (response rate 80.2%) 22 of whom had xerophthalmia, prevalence 1.32% (95% confidence interval [CI] 0.76-1.88%). There were no gender differences. Independent risk factors for xerophthalmia were having an illiterate mother [Odds ratio [OR]15.4 (95% CI 4.4-64.1)] and lack of a safe water supply [OR 6.11, 95% CI 2.5-5.1)]. Only 11.3 and 13.3% of children in different slums had taken vitamin A supplements. CONCLUSIONS: Xerophthalmia was at a level of public health significance in children living in slums in Pune. This study highlights the importance of female education and indicates that vitamin A supplementation and other approaches to control need to be improved in children living in deprived areas like urban slums. PMID- 19191177 TI - Prevalence of and associated factors for dry eye in a Spanish adult population (the Salnes Eye Study). AB - PURPOSE: To estimate the prevalence of dry eye and to investigate its relationship with lifestyle and systemic factors in a general adult population in north-western Spain. METHODS: A dry eye questionnaire was administered and objective tests were performed in 654 individuals [mean age (Standard deviation): 63.6 (14.4) years, range: 40-96, 37.2% males]. Subjects were considered symptomatic when one or more of the symptoms of the questionnaire were present often or all the time. Schirmer test or= 3 and fluorescein staining >or= 1 were considered indicative of signs. Dry eye was defined as the simultaneous presence of symptoms and at least one sign. A design based analysis was performed and all calculations were weighted to give unbiased estimates. RESULTS: Dry eye prevalence was 11.0% (95%confidence interval [CI] 8.6-13.3). Dry eye was found to be more frequent in women (11.9%, 95%CI 8.8-15.1) than in men (9.0%, 95%CI 5.3-12.6), and was significantly associated with aging (p < 0.001). After controlling for age and sex, acne rosacea was the only factor associated with the disease. There were no habits or systemic factors associated with symptoms. However, autoimmune diseases, acne rosacea, and computer use were found to be independently and significantly associated with signs. CONCLUSIONS: Dry eye is a common disease in this adult European population and is more frequent in older subjects. Acne rosacea is the only factor associated with dry eye. Other factors are associated with signs but not with symptoms. PMID- 19191178 TI - United States cost-effectiveness study of two dry eye ophthalmic lubricants. AB - PURPOSE: Dry eye is thought to be the most common ocular problem in the United States. However, it is only recently that it has been recognized as a disease with significant economic burden and impact on quality of life. This article presents an economic model of cost-effectiveness of two ophthalmic lubricants commonly used in the treatment of dry eye-Systane and Refresh Tears. METHOD: We conducted a meta-analysis, pooling the results of two clinical trials of patients with dry eye treated with Systane and Refresh Tears which presented response rates for improvements in ocular dryness, the key symptom of dry eye. An economic model was used to estimate the incremental cost-effectiveness ratio (ICER) for Systane versus Refresh Tears over a 12 month treatment period. RESULTS: Systane is significantly more effective than Refresh Tears at improving symptoms of "dryness" (75% vs 41%). Systane costs on average $57.79 per year more than Refresh Tears. Assigning a quality-adjusted life year (QALY) gain of 0.03 to responders results in an incremental cost per QALY gain of $5,837. CONCLUSION: Both Refresh Tears and Systane are effective and cost-effective interventions. Systane costs more than Refresh Tears, however clinical trial evidence shows it to be more effective. The ICER for Systane versus Refresh Tears is well below the generally accepted $50,000 per QALY threshold. PMID- 19191179 TI - A questionnaire-based assessment of symptoms associated with tear film dysfunction and lid margin disease in an Asian population. AB - PURPOSE: To study the prevalence, associated factors and impact of symptomatic dry eye in an Asian population. METHODS: A population based survey of eye diseases in 3,280 (78.7% response rate) Malay persons aged 40-80 selected from designated areas in southwestern Singapore. Participants were administered a standardized dry eye questionnaire consisting of 6 questions on symptoms, and had a comprehensive systemic and ocular examination. Symptoms of tear film dysfunction (STFD) was defined as one or more self-reported symptoms that were frequently present (ranked often or all the time). RESULTS: The prevalence rate of STFD was 6.5% (95% confidence interval [CI]: 5.7, 7.4). The prevalence was significantly higher in men compared to women (8.2% and 4.9%, respectively; p < 0.001) and decreased with age in men (p = 0.002) but not in women (p = 0.101). After adjusting for age, gender, nature of work (outdoor/indoor), and housing type, factors significantly associated with STFD were cigarette smoking (odds ratio [OR] 1.77, 95% CI: 1.17-2.66), thyroid disease (OR 2.58; 95% CI: 1.29-5.18) and higher income (OR 1.74; 95% CI: 1.13-2.68). STFD were associated with self reported difficulty in performing daily activities (work, family and leisure related) (p = 0.006). CONCLUSION: STFD are present in 6.5% of Singapore Malay adults and is associated with cigarette smoking, presence of thyroid disease and a reduced visual function-related quality of life. PMID- 19191180 TI - Prospective, multicenter demographic and epidemiological study on vernal keratoconjunctivitis: a glimpse of ocular surface in Italian population. AB - PURPOSE: To evaluate the frequency and epidemiological features of vernal keratoconjunctivitis (VKC) in Italy. METHODS: a specific electronic clinical chart for vernal keratoconjunctivitis was created to standardize: 1) medical history; 2) diagnostic criteria; 3) signs and symptoms; and 4) treatments. This study involved 6 Italian referral centers for ocular surface diseases: between March 2005 and March 2006, all referred patients were included, clinical data collected and statistically examined. RESULTS: The mean age of the vernal keratoconjunctivitis population (n = 156) was 13.8 +/- 8.8 with 64.1% of subjects under 14 years of age and a male/female ratio of 3.5:1. Among VKC patients, 48.7% showed associated systemic allergic diseases. Only 32.1% of patients were positive for RAST and/or prick test. The limbal form (53.8%) was the most frequent subtype of vernal keratoconjunctivitis. Approximately 9% of patients showed a severe form of vernal keratoconjunctivitis. At the first visit patients were treated with: multiple action or mast cell stabilizer eye drops (58.1% and 41.3% of cases, respectively), topical corticosteroids alone (0.6%) or in association (26.8% of cases). All patients used topical steroids at least once in the studied year. Systemic antihistamine therapy was used by 25.6% of patients. In this cohort, 32.7% of patients required two or more examinations per year for exacerbations of their symptoms. CONCLUSION: Vernal keratoconjunctivitis is a severe ocular condition that mainly affects young males. Vernal keratoconjunctivitis is characterized by different clinical features and therapeutic responses, suggesting the need for a standardized therapeutic approach on the basis of a grading of disease severity. PMID- 19191181 TI - Trends in reported visual impairment in United States adults. AB - The purpose of this study was to assess if rates of reported visual impairment (VI) in the United States (US) are declining. The 1997-2005 National Health Interview Survey is an annual probability survey of US households (n = 289,442 adults 18 years of age and older). Participants responding yes to either question were classified as visually impaired: 1) "Do you have any trouble seeing, even when wearing glasses or contact lenses?" 2) "Are you blind or unable to see at all?" Prevalence rates were adjusted for survey design; weighted linear regression models were used to examine VI trends. Annual rates of any VI ranged from 8.6% to 10.0% with a non-significant annual downward trend of -0.04%. Significant downward trends were noted in adults who reported an eye care provider contact in the previous 12 months (-0.18%; [standard error = 0.08], p = 0.047), adults with diabetes (-0.81%; [standard error = 0.30], p = 0.01) and adults 80 years of age and older (-0.46% [0.19]; p = 0.021). Pooled analyses with adjustment for sociodemographic and diabetes status suggested that reductions in VI tended to occur in adults with eye care provider contacts. Downward trends in reported VI in older adults may be due to improvements in the treatment of ocular disease, increased eye care utilization in those with VI, or both. These findings require verification in population-based studies with clinical acuity measures in order to specifically assess the effect of eye care utilization on VI prevalence trends. PMID- 19191182 TI - Medical care cost of Medicare/Medicaid beneficiaries with vision loss. AB - PURPOSE: To assess the impact of vision loss on healthcare cost for patients with Medicaid and Medicare and whether these costs are adequately captured by Medicare hierarchical condition categories (HCC) risk adjustment methodology. DATA SOURCES: The public use data set of the Program of All-Inclusive Care for the Elderly (PACE) for 1994-1998, and the Medicare 5% Sample datasets for 2003 and 2004. METHODS: For the first analysis, up to five years of PACE data for each individual was used to calculate HCC scores (n = 3,459). For the second analysis, claim or encounter data from Medicare Fee-for-Service (FFS) and Medicare Advantage (MA) were used to estimate the cost for each beneficiary in the upcoming payment year (n = 2,108). RESULTS: The increase in medical cost risk overall for visually impaired PACE participants was 10%, increasing to 13% for the non-institutionalized, community-based cohort, but PACE participants in nursing homes with vision loss did not generally result in increased costs. In the Medicare 5% sample, the HCC model under-predicts costs by about 17%. CONCLUSION: Our analyses provide evidence that healthcare cost risk attributable to vision loss is not adequately captured by Medicare HCC risk adjustment methodology. We hypothesize this is due to additional morbidity and treatment patterns associated with visual impairment. PMID- 19191183 TI - Cataract remains an important cause of blindness in Campinas, Brazil. AB - OBJECTIVE: To estimate the prevalence of blindness in the elderly population of Campinas, Brazil, and to describe the coverage and quality of cataract surgery services in the area. METHODS: A brief assessment of cataract surgery services (using the "RACSS" (Rapid Assessment of Cataract Surgical Services Method) was conducted using random cluster sampling, with a sample composed of 60 clusters of 40 people aged 50 years or older. Visual acuity (VA) was measured and the lens status observed by direct visual ophthalmoscopy. From the selected sample of 2,400 subjects, 92.67% were examined. RESULTS: Blindness (VA < 3/60 with available correction) was found in 1.98 % (2.03 % among male subjects, and 1.94 % among female subjects). The prevalence of blindness varied with age, from 0.2%, in the group from 50 to 54 years, to 7.2% in those above 80. Cataract was the main cause of blindness (40.2%) followed by suspected posterior segment disorders (18.2%), diabetic retinopathy (15.9%), and glaucoma (11.4%). The cataract surgical coverage was of 93% (VA < 3/60) and 82.18% when the criterion was VA 6/60 in the best eye. The main reasons the subjects did not receive surgical treatment were: fear of undergoing surgery, 11.1%; lack of awareness about the condition, 16.7%; waiting for maturity, 16.7%; and contraindication to surgery, 44.4%. CONCLUSION: Cataract is the major cause of blindness in Campinas. Education on eye diseases, their prevention and treatment must become part of the city's public healthcare policies. PMID- 19191184 TI - Effect of intravenous albumin on endotoxin removal, cytokines, and nitric oxide production in patients with cirrhosis and spontaneous bacterial peritonitis. AB - OBJECTIVE: In patients with cirrhosis and spontaneous bacterial peritonitis (SBP), the use of intravenous albumin has been shown to prevent deterioration of renal function and to decrease the mortality rate, but the mechanisms remain unclear. The purpose of this study was to characterize the mechanisms of action of albumin with the focus on endotoxin and cytokines. MATERIAL AND METHODS: Thirty patients with SBP were divided into two groups. Group 1 received antibiotics and albumin infusion (20% 50 cc every day for 3 days) and Group 2 received antibiotic treatment only. Twenty-four cirrhotic patients with sterile ascites were enrolled in Group 3 and received albumin infusion (20% 50 cc every day for 3 days). Plasma and ascitic fluid concentrations of endotoxin, nitric oxide products (NOx), tumor necrosis factor (TNF)-alpha, and interleukin (IL)-6 were analyzed before and after treatments, respectively. RESULTS: Combination therapy of albumin and antibiotics can significantly (p<0.01) reduce plasma levels of TNF-alpha and IL-6, and ascitic fluid levels of endotoxin, TNF-alpha and IL-6 in cirrhotic patients with SBP. Without the addition of albumin to an antibiotic regimen, the plasma and ascitic fluid levels of NOx increased significantly in patients with SBP (p=0.005 and p=0.004, respectively). CONCLUSIONS: The results confirm that the beneficial effects of albumin are related to the reduction of the levels of TNF-alpha and NOx in both plasma and ascitic fluid. The infusion of albumin continuously for 3 days in addition to antibiotic treatment at the time of SBP detection is recommended as an effective therapy for patients with cirrhosis and SBP. PMID- 19191185 TI - Effect of oral lansoprazole on intragastric pH after endoscopic treatment for bleeding peptic ulcer. PMID- 19191186 TI - Function and dysfunction of the colon and anorectum in adults: working team report of the Swedish Motility Group (SMoG). AB - Symptoms of fecal incontinence and constipation are common in the general population. These can, however, be unreliably reported and are poorly discriminatory for underlying pathophysiology. Furthermore, both symptoms may coexist. In the elderly, fecal impaction always must be excluded. For patients with constipation, colon transit studies, anorectal manometry and defecography may help to identify patients with slow-transit constipation and/or pelvic floor dysfunction. The best documented medical treatments for constipation are the macrogols, lactulose and isphagula. Evolving drugs include lubiprostone, which enhances colonic secretion by activating chloride channels. Surgery is restricted for a highly selected group of patients with severe slow-transit constipation and for those with large rectoceles that demonstrably cause rectal evacuatory impairment. For patients with fecal incontinence that does not resolve on antidiarrheal treatment, functional and structural evaluation with anorectal manometry and endoanal ultrasound or magnetic resonance (MR) of the anal canal may help to guide management. Sacral nerve stimulation is a rapidly evolving alternative when other treatments such as biofeedback and direct sphincter repair have failed. Advances in understanding the pathophysiology as a guide to treatment of patients with constipation and fecal incontinence is a continuing important goal for translational research. The content of this article is a summary of presentations given by the authors at the Fourth Meeting of the Swedish Motility Group, held in Gothenburg in April 2007. PMID- 19191187 TI - Combination treatment with leflunomide and methotrexate for patients with active rheumatoid arthritis. AB - OBJECTIVE: To determine the efficacy and safety of the combination of leflunomide and methotrexate for the treatment of patients with active rheumatoid arthritis (RA) in an open, non-comparative, multicentre trial. METHODS: Seventy-four patients with active RA were enrolled to receive concomitantly leflunomide (no loading dose, 10 mg/day) and methotrexate (starting at 7.5 mg/week and titrating up to 15 mg/week) for 20 weeks. The primary end-point was a 20% improvement in the American College of Rheumatology (ACR) criteria at 20 weeks. Safety measures included evaluation of adverse events at each visit and laboratory data, including haematology and liver function tests. Intention-to-treat analyses were conducted. RESULTS: Sixty-five patients completed 20 weeks of treatment, and 71.6% were responders based on the ACR20 criteria. After 20 weeks, the mean changes were -16.3 for tender joint count, -12.0 for swollen joint count, -44.0 for physician global assessment, -34.3 for patient global assessment, -22.7 for erythrocyte sedimentation rate, and -0.65 for the Health Assessment Questionnaire score. Adverse events occurred in 40.5% of the patients, and were considered serious in four patients who discontinued therapy. Abnormal liver function was noted for 16 patients (21.6%). Two of these patients were withdrawn from the study; after discontinuing the medication, their liver function recovered fully. CONCLUSION: THE combination of leflunomide and methotrexate was effective and well tolerated in the treatment of active RA patients. This combination may be a useful option as an initial treatment for active RA before starting biological agents. PMID- 19191188 TI - Lupus anticoagulant and IgM anti-phospholipid antibodies in Korean children with Henoch-Schonlein purpura. PMID- 19191190 TI - Prevalence of spondyloarthropathy in Lithuania: comment on the article by Adomaviciute et al. PMID- 19191194 TI - Whose loss is it? Human electrophysiological correlates of non-self reward processing. AB - To recognize whether another person's action results in a good or bad outcome is imperative for social learning, as well as for understanding the behavior of others in a broad context. Recent studies have reported that a scalp-surface event-related potential (ERP) called medial-frontal negativity (MFN), considered to be an index of negative reward processing, is generated when perceiving not only one's own losses, but also those of others. This suggests that the same neural mechanisms operate in monitoring one's own actions and in perceiving the consequences of the actions of others. To further elucidate the properties of this "observational" MFN, this study examined whether its amplitude differs with different observational targets. In a gambling task, participants observed the performances of non-self agents: a human friend and PC programs. The outcomes of the decisions of these agents were not associated with the participants' own benefits. ERP results showed that the MFN-like pattern was significantly elicited only when observing the outcomes of decisions made by human agents. Furthermore, self-reported measures of empathy were positively associated with the magnitude of the observational MFN. These findings suggest that the neural activity in non self reward processing reflects a socioemotional state generated by the target of observation, as well as an empathetic trait of the individual. PMID- 19191195 TI - Ultrastructural observations on inflammatory angiogenesis in gastric carcinomas with massive neutrophil infiltration. AB - Neutrophils are traditionally thought of as terminal effectors of inflammatory reaction, but experimental studies suggest that they play a direct role in the inflammatory angiogenesis of tumors. Thus, further evidence in humans is required regarding the mechanisms by which neutrophils induce tumor angiogenesis. In this study, 4 cases of human gastric carcinomas with massive neutrophil infiltration were studied by light and electron microscopy, focusing on the inflammatory angiogenesis in the tumor stroma. At light microscopy, the tumors were advanced gastric carcinomas in which various degrees of tubular differentiation were present. Under an electron microscope, pericytes exhibited two major differentiated states with distinct ultrastructural features: a contractile phenotype and a synthetic phenotype. The contractile phenotype was characterized by abundant microfilaments. Synthetic pericytes contained abundant rough endoplasmic reticulum, lipid bodies, and numerous membrane-bound vesicles. These ultrastructural findings extend concept of contractile/synthetic phenotype modulation, originally described in smooth muscle cells, to tumor microvascular pericytes. Tumor microvasculature was also characterized by abortive or slit-like lumina, endothelial cell mitoses, and replicating basement membranes. These qualitative and observational transmission electron microscopy findings provide additional morphological evidence of active inflammatory angiogenesis in gastric carcinomas with massive neutrophil infiltration. PMID- 19191196 TI - Ultrastructural features of lung fibroblast differentiation into myofibroblasts. AB - Fibroblast differentiation into myofibroblast in transforming growth factor-beta1 exposed human lung fibroblasts and the immunolocalizations of alpha-smooth muscle actin, fibronectin, tenascin-C, and osteopontin in exposed cells were studied by conventional transmission electron microscopy and immunoelectron microscopy. Ultrastructural features of myofibroblasts were detected after exposure, e.g., alpha-smooth muscle actin positive bundles in the cytoplasm of cells and extracellular fibronectin-containing structures on the surface of the cell forming fibronexus structure, osteopontin adjacent to rough endoplastic reticulum and extracellular tenascin-C in the vicinity of the cell. The authors concluded that exposure to transforming growth factor-beta1 can differentiate lung fibroblasts into ultrastructurally typical myofibroblasts. PMID- 19191197 TI - A structure resembling basal/external lamina on the surface of plasma cells, and a discussion on intercellular contacts between hemolymphoid cells. AB - Basal/external lamina is not found over plasma cells or other hemolymphoid cells, and the feature can have diagnostic value in distinguishing the neoplastic counterparts of such cells from epithelium, endothelium, mesothelium, and so on, which do have this feature. In this paper, a material ultrastructurally indistinguishable from basal or external lamina is reported on reactive plasma cells found in a fibrous pseudotumor, intralobular stroma of normal breast, tumor stroma of squamous cell carcinoma, and submucosa of normal human small intestine. It was focal, followed the contours of the cell-surface membrane, was lightly textured, 40-80 nm thick, and separated from the plasma cell surface membrane by a clear space resembling a lamina lucida. Its function remains uncertain, although it may implement adhesion to other cells, as part of the immune functions that plasma cells and other hemolymphoid cells perform in the gut, for example, and elsewhere. PMID- 19191198 TI - HCV has transforming potential to retrovirus: an ultrastructure hypothesis. AB - The present work highlights intracellular viral morphogenesis and virus-host cell interactions in patients proved to be infected with HCV. The material of this study consisted of 28 liver biopsies taken from patients positive for serum HCV RNA by polymerase chain reaction. Liver biopsies were processed for light and electron microscopic examination. Ultrastructural findings of this work supported a new hypothesis for the turnover of HCV to retrovirus and described the presumed involved mechanism. This novel perception offers important insights that can explain the vague mechanisms of HCV behavior in the infected hepatocytes. PMID- 19191199 TI - Fibronexus junctions associated with in vivo human endothelium. AB - The fibronexus is recognized as a characteristic marker of the myofibroblast. However, it is not completely specific for this cell, having been seen in aortic smooth muscle (in attenuated form) and endothelium in experimental animals. This paper documents fibronexus junctions in human in vivo endothelium. Ultrastructural observations were made on the vasculature of a desmoplastic and focally neurotropic malignant melanoma. Cross-sectioned fibronectin fibrils were seen outside the stromal surface of the endothelial plasmalemma. Often, they were positioned directly opposite the actin-filament bundles in the peripheral cytoplasm. Neoplastic and in vitro cultured cells apart, endothelium is the only nonmyofibroblastic cell type to show well-developed fibronexus junctions. Mostly, they have been documented in aortic endothelium in experimental animals, where they possibly constitute an adaptation to hemodynamic stress, and where they might more securely anchor endothelium on to subjacent connective tissue. They might also function as mechanotransducers of extracellular stress in the extracellular milieu. The present observations constitute a further, rare example of endothelium-associated fibronexuses in reactive human vessels. PMID- 19191200 TI - Carcinoma versus cytokeratin-positive lymphoma: a case report emphasizing the diagnostic role of electron microscopy. AB - Lymphoma diagnosis rarely needs electron microscopy (EM), but one area where it can be useful is in the distinction of cytokeratin-positive lymphoma from carcinoma. The authors describe such a case, where difficulties were encountered due to lack of antibody specificity, distinguishing reactive from tumoral cells, and suboptimal sampling for EM. The tumor was in a lymph node next to the right submandibular gland in a 69-year-old man. This was a malignant tumor, composed of sheets of monomorphic large round cells. Interpretation on the part of a team of pathologists who examined this tumor was divided. On histological sections, the differential diagnosis was between carcinoma and lymphoma, which was modified to cytokeratin-positive lymphoma versus carcinoma since tumor cells were found to be cytokeratin-positive. EM of tumor retrieved from formalin showed plasmablastic features, in keeping with lymphoma with plasmablastic differentiation, one of the light microscopy diagnoses. The moderately strong positivity of cytokeratin and the positivity for Ber-EP4, however, supported carcinoma, and further sampling for EM was carried out, specifically on a cytokeratin-positive area of the wax block. Tonofibrils were found, supporting carcinoma. The final diagnosis was undifferentiated carcinoma with unknown primary site. The study emphasizes the need to take into account the imperfect specificity of cytokeratin, which can be found in several hemolymphoid neoplasms, to distinguish reactive from neoplastic cells, and to secure appropriate sampling for EM. This is one of the occasions where dewaxing (of an immunohistochemically defined wax block) offers positive advantages, despite compromised structural preservation, in the search for diagnostically important organelles. PMID- 19191202 TI - Evaluation of clip artefacts in three-dimensional computed tomography. AB - OBJECTIVE: Recently, three-dimensional CT angiography (3D-CTA) has been used as a diagnostic method in postoperative evaluation after aneurysm clipping. A systematic analysis of possible predictors for good image quality needs to be conducted if 3D-CTA is to replace digital subtraction angiography in the postoperative setting. The aim of this study was to evaluate the influence of clip orientation on image quality and to assess titanium clip artefacts quantitatively on 3D-CTA. MATERIAL AND METHOD: Four different Yasargil titanium clips were placed on an acrylic glass tablet. Standardised 3D-CTA scanning was performed at clip-gantry angles of 0 degrees , 45 degrees and 90 degrees . Multiplanar reconstructions of the clips in frontal and lateral projections were performed. 3D-CTA image quality and the severity of titanium artefacts were judged by four observers. RESULTS: Twenty-four images were evaluated. Using frontal reconstructions, clip artefacts at a clip-gantry angle of 0 degrees were almost exclusively distributed along the long axis of the clip, with excellent evaluation quality at the lateral side of the clip branches. With decreasing clip gantry angle (90 degrees and 45 degrees ) strong artefacts were localised laterally. On lateral reconstructions image quality was not significantly more restricted, regardless of the clip-gantry angles used. CONCLUSION: 3D-CTA image quality can be severely limited by clip artefacts. The expression and distribution of these artefacts depends on both the clip-gantry angle and the plane of image reconstruction. Overall, there is no correlation between clip gantry angle and either the severity or the spatial distribution of clip artefacts leading to observable restrictions in the evaluation of surrounding structures. PMID- 19191203 TI - Analysis of reoperations after surgical treatment of degenerative cervical spine disorders: a report on 900 cases. AB - Surgery on the degenerative cervical spine disorders aims at decompression of the neural structures and restoring the physiological profile of the cervical spine. The aims of internal fixation are to gain primary stability, introduce a bony fusion and to correct the shape of the spine. The present study will give answers to the following questions: 1. What is the overall revision rate following an operative treatment of degenerative cervical disorders using common operative techniques? 2. Is there any influence of the fusion length to the overall revision rate and especially to the decompensation ratio of adjacent segments? 3. What is the rate of revisions due to instrumentation failures? 4. Are there any differences concerning the revision rate between posterior and anterior instrumentation? MATERIAL: We reviewed 900 patients, who underwent a cervical spine surgery with an internal fixation between January 1994 and December 2000. METHODS: Five different operative techniques were used: type I (mono-and bisegmental intersomatic decompression and fusion using anterior instrumentation), type II (multisegmental intersomatic decompression and fusion using anterior instrumentation), type III (multisegmental anterior intersomatic decompression and fusion with posterior instrumentation), type IV (one-level corpectomy with vertebral body replacement and anterior instrumentation) and type V (multi-level corpectomy with vertebral body replacement and posterior instrumentation). The minimum follow up period was 2.2 years (mean 4.2 years). RESULTS: In total, 121 revisions (13.4%) were recorded. The main indication for revision was implant failure in 5.4%. Operations type I showed the lowest revision rate (11%), while type V operations showed the highest revision rate (32%). CONCLUSIONS: The influence of the fusion length on the revision rate was unexpectedly high. Adjacent level decompensation was neither influenced by the length of the fusion nor the performed procedure. Compared to anterior instrumentation, posterior instrumentation showed a tendency for a lower revision rate without statistical significance. However, the posterior procedures showed a high revision rate regarding to wound healing problems. PMID- 19191204 TI - Fiber tracking with distinct software tools results in a clear diversity in anatomical fiber tract portrayal. AB - BACKGROUND: Fiber tract portrayal, based on diffusion tensor imaging (DTI), is becoming more and more important in functional neuronavigation. No standard exists to guarantee anatomically correct fiber tract depiction for neurosurgical purposes. Therefore, showing the anatomically correct extension of fiber tracts beyond the pure connection of functional areas remains an area of important research and investigation. Standards for fiber tracking software applications are elusive. The purpose of this study was to compare the performance of different fiber tracking software tools (FT-tools). We tested the software performance, comparability and anatomical accuracy of the tracking results of several programs. MATERIAL AND METHODS: A single DTI dataset of a healthy control subject was submitted to four different fiber tracking software applications (two commercial, two freeware), three of them based on Fiber Assignment by Continuous Tracking, one based on the Tensorline Propagation Algorithm. The corticospinal tract (CST) was investigated. The tracking procedure was controlled by the following input variables: single regions of interest (ROIs): brain stem, or internal capsule, or subcortical white matter of the precentral gyrus; background threshold, fractional anisotropy (FA) threshold, maximum fiber angulation and fiber length. Tracking results were compared for 2-D correlated triplanar images (axial, coronal, sagittal) and in 3-D. For all FT-tools, the time used to generate the CST was measured. The inter-rater variability for tracking time and for the tracked CST volumes was recorded for two of the four FT-tools. RESULTS AND CONCLUSIONS: Distinct FT-tools performed very differently with respect to the time required to achieve CST portrayal (track generation time varied between 16 and 50 min). None of the software applications was able to display the CST in its full anatomical extent. Especially the lateral precentral areas were not pictured. Surprisingly, the application of the four distinct FT-tools did not lead to comparable tracking results. As very similar or identical tracking algorithms were used, this difference cannot be easily explained. Clearly, neurosurgeons have to be cautious about applying fiber tracking results intraoperatively, especially when dealing with an abnormal or distorted fiber tract anatomy. The authors recommend the use of adjunct strategies such as intraoperative electrophysiology to enhance patient safety and improve anatomical accuracy when using tracking results for surgical procedures. PMID- 19191205 TI - Spontaneous resolution of a small extradural haematoma in a child. AB - We report the rapid spontaneous resolution of a small extradural haematoma (EDH) in a 19-month-old child. The haematoma was managed conservatively. The child went on to make a full recovery. The authors discuss the significance of this surprising clinical finding. PMID- 19191206 TI - Multifocal glioblastoma with remote cutaneous metastasis: a case report and review of the literature. AB - BACKGROUND: Remote extracranial mestastases of glioblastoma multiforme (GBM) are uncommon, while cutaneous seeding at a distance from the operative site appears to be even more unusual. CASE REPORT: A 63-year-old man presented with focal seizures and mental impairment. Computed tomography (CT) scan revealed a left frontoparietal mass. He underwent a gross total removal of the tumor. The tissue diagnosis was that of a GBM. Seven months later, the patient developed a left scapular subcutaneous mass. Fine-needle aspiration cytology (FNAC) was performed and the cytological findings disclosed again a GBM. One month later, after clinical deterioration, a repeat magnetic resonance imaging (MRI) scan was carried out which demonstrated two new distinct lesions in the opposite hemisphere, as in a multifocal GBM. Both lesions were biopsed under stereotactic guidance and the recurrence of GBM was confirmed. The patient died ten months after the primary diagnosis of the intracranial GBM. CONCLUSION: Improved diagnostic modalities and prolonged survival have increased the likelihood of detection of extracranial mestastases from GBM. This potential may be greater in multifocal GBM. FNA is a valuable method for the definite diagnosis of metastatic GBMs. Although several theories have been postulated, the route of remote cutaneous dissemination and the mechanism of multifocal recurrence remain to be elucidated. PMID- 19191207 TI - A non-midline spheno-orbital encephalocele in a newborn. AB - Basal encephaloceles in western countries occur in 1 of every 35 000-40 000 live births; with an incidence of less than 10% they are the least common of all encephaloceles. Certain subtypes such as transsphenoidal variants may be as rare as 1 in 700 000 live births. These rare encephaloceles are classified into five anatomic types: spheno-ethmodial, transsphenoidal, spheno-orbital, transethmoidal, and spheno-maxillary. Here we present an exceedingly rare variant of a non-midline basal encephalocele of the spheno-orbital type, which was treated by resection of the encephalocele, which contained dysplastic central nervous system tissue, on day four post partum. The patient had no neurological deficits and a six year follow-up showed a normal intellect and a good cosmetic result. PMID- 19191208 TI - Levosimendan does not improve survival time in a rat model of verapamil toxicity. AB - OBJECTIVE: Calcium channel blocker (CCB) toxicity, in particular that induced by verapamil and diltiazem, presents clinical challenges with no true antidote. Levosimendan, a calcium sensitizer, improves cardiac contractility in patients with heart failure. We tested the hypothesis that calcium channel sensitization will prolong survival in a rat model of severe verapamil poisoning. METHODS: This was a blinded, randomized, controlled animal study. Wistar rats (mean weight, 371 +/- 50 g) were used. Verapamil (2.5 mg/ml) was infused at a rate of 37.5 mg/kg per hour. Bolus doses of levosimendan (5 microg/mL) were given at 0 min (12 microg/kg) and 5 min (18 microg/kg); saline control was of equal volume. The rats were intubated and maintained under general anesthesia with isoflurane. Electrocardiographic activity and core temperature were monitored during the poisoning and treatment phases. Each rat underwent femoral vein cannulation and was then randomized, in blinded fashion, to receive either levosimendan or an equal volume of saline at 0 and 5 minutes. Death, defined as 1 minute of asystole, was used as the primary endpoint. RESULTS: Rats treated with levosimendan died before the control group (7.37 +/- 0.7 min [n=7] vs. 16.4 +/- 4.2 [n=7] [ p=.053]). All animals experienced bradycardia prior to asystole. DISCUSSION: Although levosimendan has the ability to sensitize and enhance binding of troponin C to Ca2+, this study did not show an improvement in survival time in the setting of verapamil toxicity. This may be attributed to levosimendan's inhibition of phosphodiesterase, which possibly exacerbated the CCB-induced hypotension. CONCLUSION: In this rat model, levosimendan as a solitary antidotal treatment for verapamil toxicity was not beneficial. PMID- 19191209 TI - Racial variations in the incidence of severe alcohol withdrawal. AB - The use of race as a risk assessment tool and pharmacologic target has garnered recent attention and debate. It is currently unclear if a relationship between race and the development of severe alcohol withdrawal exists. We explored this potential relationship using several study groups. METHODS: A simultaneous prospective enrollment of patients and retrospective chart review of severe alcohol withdrawal in two separate settings was performed comparing both the incidence of withdrawal and alcoholism based on race. These two study groups were then compared to an "at risk" group of alcoholics and the general ED population to determine differences in the distribution of race. RESULTS: Individuals of white race in both study groups were at increased odds [OR 1.93 (CI 1.11-3.39) and 2.19 (CI 1.41-3.40)] of having severe alcohol withdrawal when compared to non White "at risk" alcoholics. Blacks in both study groups however, appear to have lower odds [OR 0.23 (CI 0.11-0.47) and 0.11 (CI 0.05-0.23)] of having severe alcohol withdrawal when compared to non-Black "at risk" alcoholics. CONCLUSIONS: Despite the controversial use of race in medical research and targeting therapies, there appears to be a difference in the odds of severe alcohol withdrawal based on race. The reasons for this finding are currently unclear. PMID- 19191210 TI - Observation unit experience for pediatric poison exposures. AB - BACKGROUND: Short-Stay Emergency Department Observation Units (OU) are an alternative to hospitalization, but data on OU care of pediatric poisoning exposures is limited. We report the experience of a pediatric OU with this population. METHODS: We retrospectively reviewed the charts of children with poison exposure admitted to a pediatric OU during a 30-month period. Data was collected pertaining to demographics, type of exposure, clinical presentation, and rate of hospitalization, and was compared to nonpoisoned OU patients. RESULTS: Of the 91 pediatric patients with poison exposure, 86 complete charts were available for review (94.5%). Of these patients, 49.5% were female, and 82.4% were <6 years of age (range 1.5 months to 16.6 years). There were a total of 98 toxicants implicated, the most common of which were psychoactive drugs (25%) and cardiovascular agents (19%). At OU admission, 33 of 88 patients (38%) had altered mental status or abnormal vital signs. Only 2 of the 53 remaining patients developed abnormal vital signs within the OU. Two patients were hospitalized unexpectedly with respiratory distress due to hydrocarbon and charcoal aspiration pneumonitis, respectively; the unexpected hospitalization rate was 2.2%. Three more planned hospitalizations for endoscopy or psychiatric evaluation led to a total hospitalization rate of 5.4%. This hospitalization rate is significantly lower (RR=0.26, 95% CI=0.11-0.62) than the hospitalization rate from the OU for nonpoisoned patients (20.3%) during that time. Mean OU length of stay for nonadmitted poisoned patients was 14.35 hours. There were no adverse events noted as a result of OU placement. CONCLUSION: Select poisoned pediatric patients appear suitable for OU management and had less frequent unexpected hospitalization from the OU than other diagnoses. PMID- 19191211 TI - Late-onset seizures associated with quetiapine poisoning. AB - INTRODUCTION: Quetiapine, a second-generation antipsychotic, acts at multiple brain neurotransmitter receptors and has the potential for serious complications. Although seizures have been described in the literature, delayed seizure onset has not been reported. We report the first case of delayed seizures after a significant quetiapine overdose. CASE REPORT: A 27-year-old female presented to the emergency department following an overdose of approximately 30 g of quetiapine. Twenty-four hours after arrival, the patient had 2 seizures. The patient was then intubated and remained in the ICU for four days. EEG was negative for epileptiform activity. The serum quetiapine levels (MedTox, St. Paul, MN) were 8.67 mg/L on hospital day one and 3.28 mg/L on hospital day three. DISCUSSION: Quetiapine poisoning, with serum levels, associated with seizures has been reported in one prior case. Our case report represents late-onset seizures with serum levels above therapeutic range (>1 mg/L). The serum concentrations of quetiapine in this case were consistent with those in postmortem case reports. PMID- 19191212 TI - Unrecognized hypoglycemia due to maltodextrin interference with bedside glucometry. AB - INTRODUCTION: Glucometry is widely used to confirm or exclude hypoglycemia in patients with suggestive clinical findings. Nonglucose sugars may be detected by certain types of glucometers, causing false elevation of the glucometer analysis of the blood sugar. Since these other sugars are not functionally glucose and may even induce excess insulin release, clinical hypoglycemia may be missed. CASE REPORT: We report a 79-year-old man on enteral feeds containing maltodextrin, a glucose polymer, who had persistently high glucometer-measured blood glucose despite normal blood glucose measured by formal laboratory analysis. DISCUSSION: Excess insulin administration, based on the erroneous glucometer reading, may have caused unrecognized fatal clinical hypoglycemia. This has been reported following intravenous administration of related nonglucose sugars but not with enteral maltodextrin. Further study is required to confirm the effects of maltodextrin on glucometry. CONCLUSION: False elevation of blood glucose measured on certain point-of-care glucometers can occur following the oral administration of maltodextrin. PMID- 19191213 TI - Neuroleptic malignant syndrome developing after acute overdose with olanzapine and chlorpromazine. AB - Neuroleptic malignant syndrome (NMS) is a relatively uncommon side effect that may develop after a recent increase in the therapeutic dose of an antipsychotic medication or the addition of a new agent in therapeutic doses. CASE REPORT: We report a case of NMS developing in a 36-year-old female patient 2 days following deliberate self-poisoning with 30 x 10-mg olanzapine tablets, 7 x 100-mg chlorpromazine tablets and an unknown amount of escitalopram. These were the patient's own medications. She had not been taking these for several weeks. The patient initially presented with sedation from her overdose which resolved over the next 24 hours. Following this, over the subsequent 24 hours, she became progressively confused, ataxic, hypertonic, ferbrile and tachycardic, with marked lead pipe rigidity of the limbs. Head CT, lumbar puncture and septic screen were all negative. She was treated with intravenous midazolam infusion, nasogastrically administered bromocriptine, external cooling and was mechanically ventilated. She gradually improved over a period of 10 days, with residual confusion lasting another week, and was discharged well with no deterioration from her premorbid neurologic state. CONCLUSION: To our knowledge, although there are numerous cases reported with therapeutic use, NMS has not been reported to develop following acute olanzapine overdose. Clinicians should be aware that this may be an uncommon side effect of antipsychotic medication. PMID- 19191214 TI - Amanita smithiana mushroom ingestion: a case of delayed renal failure and literature review. AB - INTRODUCTION: In the Pacific Northwest a new pattern of mushroom ingestion has emerged, attributed to Amanita smithiana, in which renal failure has been the predominant manifestation. CASE REPORT: A 55-year-old male ate 3 raw wild mushrooms in a salad and had onset of severe nausea and vomiting within 6 hours. His vital signs were unremarkable. His labs were significant for a BUN of 14 mg/dL (5.0 mmol/L), and a creatinine of 1.0 mg/dL (88 umol/L), transaminases were elevated with an AST of 56 U/L (nl 9-40) and an ALT of 131 U/L (nl 14-72). Treatment was initiated with N-acetyl cysteine, penicillin, and milk thistle extract on the presumption that this was an amanitin-toxin containing mushroom. He developed acute renal failure that was not responsive to our treatment. Dialysis started on day 4 with a creatinine of 6.5 mg/dL, which peaked on day 7 at 10.2 mg/dL. We were able to obtain a positive mushroom identification by a mycologist as Amanita smithiana. The patient was discharged from the hospital for outpatient dialysis on day 10 and dialysis catheter was removed 39 days after ingestion with a creatinine of 1.4 mg/dL (123.8 umol/L). DISCUSSION: Amanita smithiana mushroom poisoning presents within 6 hours of ingestion with GI toxicity, and develops delayed onset of renal insufficiency over the first 1 to 4 days. The early hospitalization of this case allowed a profile of the onset of liver and renal injury. Mild elevation of hepatic transaminases occurred on presentation and peaked 24 hours after the ingestion. Renal injury was detected 1 day after presentation, and progressed to require hemodialysis by 4 days postingestion. This pattern of delayed-onset renal toxic mushroom ingestion is emerging among mushroom ingestions in Western North America. PMID- 19191215 TI - Case files of the Medical Toxicology Fellowship at the University of Massachusetts Medical Center in Worcester, Massachusetts: stumbling through a blue haze. PMID- 19191216 TI - Medical toxicology and public health--update on research and activities at the Centers for Disease Control and Prevention, and the Agency for Toxic Substances and Disease Registry: introduction to the Laboratory Response Network-Chemical (LRN-C). PMID- 19191217 TI - Hoxb13 regulatory elements mediate transgene expression during prostate organogenesis and carcinogenesis. AB - The prostate requires androgens for development and homeostasis. Prostate cancer shares this dependence, however progression to androgen-independence is common after androgen deprivation. There is considerable interest in achieving therapeutic gene expression after androgen ablation using prostate-specific promoters. Paradoxically, known prostate-restricted cis-regulatory elements are androgen dependent. Hoxb13 expression is restricted in adults to the prostate and colon, and robust Hoxb13 expression persists after castration. To locate regulatory elements conferring this expression pattern, a lacZ reporter was inserted into the Hoxb13 locus on a mouse genomic bacterial artificial chromosome. In transgenic mice, this construct recapitulated the Hoxb13 expression pattern, including expression after castration. Reporter gene activity was maintained during carcinogenesis in a prostate cancer model. Hoxb13 cis regulatory elements provide a powerful tool to achieve androgen-independent transgene expression in the prostate and distal colon-specific expression in the gastrointestinal tract. These data establish a framework for high-resolution analyses of factors regulating Hoxb13. PMID- 19191218 TI - C-Fos elimination compensates for disabled-2 requirement in mouse extraembryonic endoderm development. AB - Disabled-2 (Dab2) is expressed in primitive endoderm cells as they are differentiating from the inner cell mass and dab2 deficiency in mice results in lethality at E5.5-E6.5 due to the disorganization of the endoderm layers. Here we show that Dab2 suppresses c-Fos expression in endoderm cells. A morphological normal primitive endoderm layer was observed in putative E5.5 dab2 (-/-):c-fos ( /-) embryos, indicating that the primitive endoderm defect due to the loss of Dab2 is rescued by deletion of the c-fos gene. The lethality of the double knockout embryos was delayed until E9.5-E10.5 and the defective embryos failed to undergo organogenesis. We conclude that Dab2 plays a role in epithelial organization by suppression of c-Fos expression and suggest that unsuppressed c Fos can lead to disruption of primitive endoderm epithelial organization, yet an additional dab2 function is required for later organogenesis. PMID- 19191220 TI - New meaning in the message: noncoding RNAs and their role in retinal development. AB - Recent studies have indicated that non-protein-coding RNAs (ncRNAs) may play prominent and diverse roles in the development of the nervous system. These ncRNAs are now known to perform a broad range of cellular functions, and in particular appear to be prominent players in the regulation of transcription and translation. In this review, we discuss recent advances in our understanding of the role of ncRNAs in vertebrate retinal development. Noncoding RNAs that are known or suspected to play a functional role in the specification and maturation of retinal cell subtypes include miRNAs, long noncoding opposite-strand transcripts (OSTs), and other long ncRNAs such as Tug1 and RNCR2. Though the mechanism of action of most of these ncRNAs is still largely unclear, it is likely that these molecules represent a major, and thus far largely unappreciated, component of the molecular machinery involved in retinal cell fate specification. PMID- 19191221 TI - Anatomical location of mature GnRH neurons corresponds with their birthdate in the developing mouse. AB - The gonadotropin-releasing hormone (GnRH) neurons exhibit substantial functional, anatomical, and molecular heterogeneity, which has hampered their thorough examination. This study was undertaken in an effort to understand whether the anatomical distribution of GnRH neurons is related to their developmental history, because such an association may help explain differences within the population. Using bromodeoxyuridine pulse labeling of timed pregnant female mice we labeled dividing cells, including GnRH neuron progenitors in the olfactory placode, throughout the window of GnRH neuron differentiation. Our results indicate that cells that become postmitotic early tend to populate the rostral aspects of the adult GnRH neuron continuum, whereas later-generated cells tend to settle more caudally; an inside-out pattern reminiscent of neocortex. These observations suggest that the timing of differentiation influences the ability of postmitotic GnRH neurons to navigate to their adult location, and hence may be important in determining the ultimate wiring of the adult network. PMID- 19191219 TI - Acheate-scute like 1 (Ascl1) is required for normal delta-like (Dll) gene expression and notch signaling during retinal development. AB - Delta gene expression in Drosophila is regulated by proneural basic helix-loop helix (bHLH) transcription factors, such as acheate-scute. In vertebrates, multiple Delta-like and proneural bHLH genes are expressed during neurogenesis, especially in the retina. We recently uncovered a relationship between Acheate scute like 1 (Ascl1), Delta-like genes, and Notch in chick retinal progenitors. Here, we report that mammalian retinal progenitors are also the primary source of Delta-like genes, likely signaling through Notch among themselves, while differentiating neurons expressed Jagged2. Ascl1 is coexpressed in Delta-like and Notch active progenitors, and required for normal Delta-like gene expression and Notch signaling. We also reveal a role for Ascl1 in the regulation of Hes6, a proneurogenic factor that inhibits Notch signaling to promote neural rather than glial differentiation. Thus, these results suggest a molecular mechanism whereby attenuated Notch levels coupled with reduced proneurogenic activity in progenitors leads to increased gliogenesis and decreased neurogenesis in the Ascl1-deficient retina. PMID- 19191223 TI - Adaptation of GAL4 activators for GAL4 enhancer trapping in zebrafish. AB - An enhancer trap-based GAL4-UAS system in zebrafish requires strong GAL4 activators with minimal adverse effects. However, the activity of yeast GAL4 is too low in zebrafish, while a fusion protein of the GAL4 DNA-binding domain and the VP16 activation domain is toxic to embryonic development, even when expressed at low levels. To alleviate this toxicity, we developed variant GAL4 activators by fusing either multimeric forms of the VP16 minimal activation domain or the NF kappaB activation domain to the GAL4 DNA-binding domain. These variant GAL4 activators are sufficiently innocuous and yet highly effective transactivators in developing zebrafish. Enhancer-trap vectors containing these GAL4 activators downstream of an appropriate weak promoter were randomly inserted into the zebrafish genome using the Sleeping Beauty transposon system. By the combination of these genetic elements, we have successfully developed enhancer trap lines that activate UAS-dependent reporter genes in a tissue-specific fashion that reflects trapped enhancer activities. PMID- 19191222 TI - Identification of embryonic pancreatic genes using Xenopus DNA microarrays. AB - The pancreas is both an exocrine and endocrine endodermal organ involved in digestion and glucose homeostasis. During embryogenesis, the anlagen of the pancreas arise from dorsal and ventral evaginations of the foregut that later fuse to form a single organ. To better understand the molecular genetics of early pancreas development, we sought to isolate markers that are uniquely expressed in this tissue. Microarray analysis was performed comparing dissected pancreatic buds, liver buds, and the stomach region of tadpole stage Xenopus embryos. A total of 912 genes were found to be differentially expressed between these organs during early stages of organogenesis. K-means clustering analysis predicted 120 of these genes to be specifically enriched in the pancreas. Of these, we report on the novel expression patterns of 24 genes. Our analyses implicate the involvement of previously unsuspected signaling pathways during early pancreas development. Developmental Dynamics 238:1455-1466, 2009. (c) 2009 Wiley-Liss, Inc. PMID- 19191224 TI - Deletion of a conserved noncoding sequence in Plzf intron leads to Plzf down regulation in limb bud and polydactyly in the rat. AB - Lx mutation in SHR.Lx rat manifests in homozygotes as hindlimb preaxial polydactyly. It was previously mapped to a chromosome 8 segment containing the Plzf gene. Plzf (promyelocytic leukemia zinc finger protein) influences limb development as a direct repressor of posterior HoxD genes. However, the Plzf coding sequence is intact in the Lx mutants. Using linkage mapping in F2 hybrids, we downsized the segment containing Lx to 155 kb and sequenced conserved noncoding elements (CNEs) inside. A 2,964-bp deletion in Plzf intron 2, never detected in control animals, is the only candidate for Lx. The deletion removes the most deeply conserved CNE in the 155-kb segment, suggesting a regulatory influence on Plzf expression. Correspondingly, using in situ hybridization and quantitative real-time polymerase chain reaction, we found a decrease of Plzf expression in Lx/Lx limb buds with concomitant anterior expansion of expression domains of its targets, Hoxd10-13 genes, in the absence of ectopic Sonic hedgehog expression. Upstream regulation of Plzf in limb buds is currently unknown. We present here the first candidate Plzf cis-regulatory sequence. PMID- 19191226 TI - Systemic hyalinosis mutations in the CMG2 ectodomain leading to loss of function through retention in the endoplasmic reticulum. AB - Systemic hyalinosis is an autosomal recessive disease that encompasses two allelic syndromes, infantile systemic hyalinosis (ISH) and juvenile hyaline fibromatosis (JHF), which are caused by mutations in the CMG2 gene. Here we have analyzed the cellular consequences of five patient-derived point mutations in the extracellular von Willebrand domain or the transmembrane domain of the CMG2 protein. We found that four of the mutations led to retention of the protein in the endoplasmic reticulum (ER), albeit through different mechanisms. Analysis of recombinant CMG2 von Willebrand factor A (vWA) domains, to which three of the mutations map, indicated that the mutations did not prevent proper folding and ligand binding, suggesting that, in vivo, slow folding, rather than misfolding, is responsible for ER retention. Our work shows that systemic hyalinosis can be qualified as a conformational disease, at least for the mutations that have been mapped to the extracellular and transmembrane domains. The long ER half-life and the ligand binding ability of the mutated von Willebrand domains suggest that treatments based on chemical chaperones could be beneficial. PMID- 19191225 TI - The Allantoic Core Domain: new insights into development of the murine allantois and its relation to the primitive streak. AB - The whereabouts and properties of the posterior end of the primitive streak have not been identified in any species. In the mouse, the streak's posterior terminus is assumed to be confined to the embryonic compartment, and to give rise to the allantois, which links the embryo to its mother during pregnancy. In this study, we have refined our understanding of the biology of the murine posterior primitive streak and its relation to the allantois. Through a combination of immunostaining and morphology, we demonstrate that the primitive streak spans the posterior extraembryonic and embryonic regions at the onset of the neural plate stage ( approximately 7.0 days postcoitum, dpc). Several hours later, the allantoic bud emerges from the extraembryonic component of the primitive streak (XPS). Then, possibly in collaboration with overlying allantois-associated extraembryonic visceral endoderm, the XPS establishes a germinal center within the allantois, named here the Allantoic Core Domain (ACD). Microsurgical removal of the ACD beyond headfold (HF) stages resulted in the formation of allantoic regenerates that lacked the ACD and failed to elongate; nevertheless, vasculogenesis and vascular patterning proceeded. In situ and transplantation fate mapping demonstrated that, from HF stages onward, the ACD's progenitor pool contributed to the allantois exclusive of the proximal flanks. By contrast, the posterior intraembryonic primitive streak (IPS) provided the flanks. Grafting the ACD into T(C)/T(C) hosts, whose allantoises are significantly foreshortened, restored allantoic elongation. These results revealed that the ACD is essential for allantoic elongation, but the cues required for vascularization lie outside of it. On the basis of these and previous findings, we conclude that the posterior primitive streak of the mouse conceptus is far more complex than was previously believed. Our results provide new directives for addressing the origin and development of the umbilical cord, and establish a novel paradigm for investigating the fetal/placental relationship. PMID- 19191227 TI - LGI1 mutations in autosomal dominant and sporadic lateral temporal epilepsy. AB - Autosomal dominant lateral temporal epilepsy (ADLTE) or autosomal dominant partial epilepsy with auditory features (ADPEAF) is an inherited epileptic syndrome with onset in childhood/adolescence and benign evolution. The hallmark of the syndrome consists of typical auditory auras or ictal aphasia in most affected family members. ADTLE/ADPEAF is associated in about half of the families with mutations of the leucine-rich, glioma-inactivated 1 (LGI1) gene. In addition, de novo LGI1 mutations are found in about 2% of sporadic cases with idiopathic partial epilepsy with auditory features, who are clinically similar to the majority of patients with ADLTE/ADPEAF but have no family history. Twenty five LGI1 mutations have been described in familial and sporadic lateral temporal epilepsy patients. The mutations are distributed throughout the gene and are mostly missense mutations occurring in both the N-terminal leucine rich repeat (LRR) and C-terminal EPTP (beta propeller) protein domains. We show a tridimensional model of the LRR protein region that allows missense mutations of this region to be divided into two distinct groups: structural and functional mutations. Frameshift, nonsense and splice site point mutations have also been reported that result in protein truncation or internal deletion. The various types of mutations are associated with a rather homogeneous phenotype, and no obvious genotype-phenotype correlation can be identified. Both truncating and missense mutations appear to prevent secretion of mutant proteins, suggesting a loss of function effect of mutations. The function of LGI1 is unclear. Several molecular mechanisms possibly leading to lateral temporal epilepsy are illustrated and briefly discussed. PMID- 19191228 TI - Physician's production of primary care in Ontario, Canada. AB - This paper examines the factors affecting the number of patient visits per week reported by family physicians in Ontario. The way that a physician is paid is potentially endogenous to the number of patients seen per week, thus an instrumental variable method of estimation is employed to account for the endogeneity bias. Once account is taken of the endogeneity of remuneration as well as relevant physician and practice characteristics, the estimated elasticity of output with respect to hours worked is 0.74; 0.68 in group practices and 0.82 in solo practices. Physicians paid on a non-fee-for-service (NFFS) conduct 15-31% fewer patient visits per week in comparison to those paid under an FFS scheme. Certain patient populations in practices affect patient visits in important ways, as do a number of physician and practice characteristics. PMID- 19191229 TI - Quantitative assessment of dielectric parameters for membrane lipid bi-layers from RF permittivity measurements. AB - In this article, we propose and validate theoretical and experimental methods to quantitatively assess the Debye dielectric model of membrane lipid bi-layers. This consists of two steps: permittivity measurements of biological solutions (liposomes), and estimation of the model parameters by inverse application of the Effective Medium Theory. The measurements are conducted in the frequency domain between 100 MHz and 2 GHz using a modified coaxial connector, at the temperatures of 27 and 30 degrees C. Estimations have been performed using a three-layered model based on the Maxwell-Wagner formulation. Debye parameters (mean value +/- standard error) found from fitting experimental data are: epsilon(s) = 11.69 +/- 0.09, epsilon(infinity) = 4.00 +/- 0.07, f(relax) = 179.85 +/- 6.20 MHz and epsilon(s) = (1.1 +/- 0.1) x 10(-7) S/m. This model can be used in microdosimetric studies aiming to precisely determine the E-field distribution in a biological target down to the single cell level. In this context the use of an accurate membrane dielectric model, valid through a wide frequency range, is particularly appropriate. PMID- 19191231 TI - Parameters influencing the release of tertiary alcohols from the surface of "spherical" dendrimers and "linear" stylomers by neighbouring-group-assisted hydrolysis of 2-carbamoylbenzoates. AB - The influence of structural and physico-chemical parameters on the release of a volatile tertiary alcohol (2-methyl-1-phenyl-2-propanol) by neighbouring-group assisted cyclisation of 2-carbamoylbenzoates at neutral pH was investigated by comparing the covalent-bond cleavage from the surface of linear, comblike poly(propylene imine) "stylomers" and their corresponding spherical, globular dendrimers. Determination of the kinetic rate constants for the stepwise intramolecular cyclisation of the 2-carbamoylbenzoate moiety by using HPLC showed that the polarity of the conjugates, and thus their solubility in the aqueous reaction medium, has a stronger influence on the rates of hydrolysis than the size (generation) or shape (linear or spherical) of the macromolecules. Furthermore, structural modifications in close proximity to the release unit, such as the presence of functionalities with catalytic activity, have a strong impact on the release efficiency of the active molecules. An understanding of the physico-chemical parameters determining the local environment of the covalent bond cleavage site is therefore an important prerequisite to transfer the characteristics of small molecules to larger structures such as oligomers and polymers and thus to design efficient macromolecular conjugates for the controlled delivery of bioactive compounds. PMID- 19191230 TI - Beyond molecular beacons: optical sensors based on the binding-induced folding of proteins and polypeptides. AB - Many polypeptides and small proteins can be readily engineered such that they only fold upon binding a specific target ligand. This approach couples target recognition with a considerable change in polymer structure and dynamics. Recent years have seen the development of a number of biosensors that couple these large changes to readily measurable optical (fluorescent) outputs. These sensors afford the detection of a wide variety of macromolecular targets including proteins, polypeptides, and nucleic acids. Here we describe the design of such biosensors, from the first iterations as protein engineering experiments, to the development of biosensors targeting a range of protein and nucleic acid targets. PMID- 19191232 TI - The quest for ring opening of oxaphosphirane complexes: a coupled-cluster and density functional study of CH(3)PO isomers and their Cr(CO)(5) complexes. AB - Opening gambit: A high-level theoretical study on the relative stabilities of oxaphosphirane isomers and their Cr(CO)(5) complexes is reported (see picture). Furthermore, thermodynamics and kinetics of possible ring-opening reactions of these complexes in the presence of a {Cp(2)Ti(III)Cl} fragment are theoretically investigated. The C--O bond cleavage is predicted to be the most efficient pathway, thus leading to reactive intermediates that are attractive for synthetic applications.A high-level theoretical study on the relative stabilities of oxaphosphirane isomers and their Cr(CO)(5) complexes is reported. Furthermore, thermodynamics and kinetics of possible ring-opening reactions of these complexes in the presence of a {Cp(2)Ti(III)Cl} fragment are theoretically investigated. The C--O bond cleavage is predicted to be the most efficient pathway thus leading to reactive intermediates that are attractive for synthetic applications. The ring-opening reaction is predicted to not lead to the most favorable product (a coordinated phosphinidene oxide species). Rather, the ring-opening product is separated by a substantial barrier of about 24 kcal mol(-1) from the thermodynamically most favorable species. PMID- 19191233 TI - Insight into the defects of cage-type silica mesoporous crystals with Fd3m symmetry: TEM observations and a new proposal of "polyhedron packing" for the crystals. AB - Silica mesoporous crystals were synthesized by using a gemini cationic surfactant (C(18-3-1)) as the directing agent, carboxyethylsilanetriol sodium salt as the co structure directing agent (CSDA), and varying amounts of HCl. By using transmission electron microscopy (TEM) we observed 1) a structural change from the close-packed structures of spherical micelles--face-centered cubic (Fm3m) and hexagonal close-packed (P6(3)/mmc)--to Fd3m structures with an increase of HCl and 2) a few structural defects in the crystals with Fd3m symmetry. The structure of a crystal with Fd3m symmetry is described as one of the tetrahedrally close packed (tcp) structures consisting of 5(12) and 5(12)6(4) polyhedra. The observed TEM images of the structural defects were explained well through use of simulated TEM images by introducing new 13-15 polyhedra comprising 5(12)6(2), 5(12)6(3), 4(1)5(10)6(2), 4(2)5(8)6(5), and 4(1)5(10)6(4), which have been observed in bubbles by Matzke. The mesostructural changes and defect formation are discussed in terms of the hardness of micelles composed of surfactant/CSDA/silica species that have formed through a change of the interaction between the surfactant and CSDA, which causes the micelles to change from a regime of close-packing to one of minimum-area packing. PMID- 19191234 TI - Two-dimensional oligo(phenylene-ethynylene-butadiynylene)s: all-covalent nanoscale spoked wheels. AB - Round and round: Covalently bound spokes induce an efficient template-directed cyclization towards a rigid molecular wheel (see figure) and afford dramatically increased shape-persistence properties compared with non-strutted macrocycles.The synthesis and characterization of a shape-persistent two-dimensional (2D) organic compound is described in detail. In a rational modular synthesis of a dodecaacetylene precursor and its subsequent template-aided cyclization, we obtained a molecularly defined, stable, C(6)-symmetric, rigid, spoked wheel. Peripheral tert-butyl groups and alkyl chains attached to the plane of the molecule provide sufficient solubility, so that the 2D oligomer can be fully characterized by MALDI-MS, GPC, and (1)H NMR, UV/Vis absorption, and fluorescence spectroscopy. Molecular mechanics and dynamics simulations indicate that the most stable conformer of the molecule in vacuum is a shallow boat conformation with a small dihedral angle. Comparisons with the precursor as well as a ring-only structure clearly reveal the high rigidity of the title compound. Small-angle neutron scattering (SANS) experiments in [D(8)]THF and CDCl(3) affirm the rigid backbone structure in solution, that is, a radius of about 2.7 nm and a thickness of about 0.22 nm. STM investigations illustrate that the wheel molecules adsorb with their molecular plane parallel to the surface and can form hexagonal crystalline domains (unit cell parameters are a=b=6.0+/-0.2 nm and theta=60+/-2 degrees ), with the tert-butyl groups on the apexes staggered. Such staggering induces chirality in the organized domains. AFM investigations demonstrate that the wheel molecules inside overlayers organize in the same way as in the layer directly in contact with the surface. This indicates an epitaxial growth characteristic of the film. PMID- 19191235 TI - New polyalkynyl dendrons and dendrimers: "click" chemistry with azidomethylferrocene and specific anion and cation electrochemical sensing properties of the 1,2,3-triazole-containing dendrimers. AB - The synthesis and use of the new tris-alkynyl dendrons 2 to 5 are reported, including the Williamson reaction of 5 with 9-iodo (9), 27-iodo (11), and 81-iodo (12) dendritic cores to yield 27-alkynyl (13), 81-alkynyl (14), and 243-alkynyl (15) dendrimers. So-called "click" reactions of these three dendrimers with azidomethylferrocene (20) give 27-ferrocenyl (16), 81-ferrocenyl (17), and 243 ferrocenyl (18) dendrimers. Electrochemical recognition of oxo-anions (H(2)PO(4)( ) and ATP(2-)) and Pd(2+) cation has been compared using the three polyferrocenyl dendrimers. Derivatization of Pt electrodes with the dendrimers for recognition becomes more facile with increasing size of the dendrimer. This first "click" dendrimer bearing 243-ferrocenyl groups is the best one in the series to obtain robust, recyclable modified Pt electrodes, whereas previous "click" ferrocenyl dendrimers have not been suitable for this purpose. PMID- 19191236 TI - Selectively N-protected enantiopure 2,5-disubstituted piperazines: avoiding the pitfalls in solid-phase Fukuyama-Mitsunobu cyclizations. AB - An efficient solid-phase route to ring-substituted piperazines from O-linked resin-bound (S)-aziridine-2-methanol is described. Regioselective microwave assisted aminolysis followed by intramolecular Fukuyama-Mitsunobu cyclization constitute the key features of the protocol. Simple piperazines and diazepanes were readily obtained without preceding N-protection of the acyclic intermediate, whereas attempts to extend this protocol to chiral 2,5-disubstituted piperazines failed. Modifications encompassing N-carbamoylation prior to ring-closure were therefore investigated. However, standard carbamoylating agents, for example, Fmoc-Cl and Alloc-Cl tended to give bis-protected by-products. Thus, novel microwave-assisted solid-phase N-protection procedures were developed for efficient introduction of Fmoc, Boc and Alloc groups. The subsequent cyclization proceeded in moderate to excellent yields depending on the bulk of the side chain and type of N-protecting group. This protocol readily provided novel cis- and trans-2,5-disubstituted piperazines displaying a variety of N-protecting group patterns after further on-resin manipulations. Also, unexpected by-products obtained during these optimization studies were identified and characterized. This includes nosylated ureas arising from an alternative cyclization pathway. Finally, post-cleavage oxidation gave access to the Fmoc/Boc-protected alpha amino acid as well as the corresponding aldehyde. The chiral piperazines described in this work will enable construction of combinatorial libraries with a higher chemical diversity compared to those containing simple N,N' difunctionalized piperazines, often present in drug-like compounds. PMID- 19191237 TI - Palladium-catalyzed cyclopropanation of alkenyl silanes by diazoalkanes: evidence for a Pd(0) mechanism. AB - Alkenyl silanes are efficiently converted to the corresponding silyl cyclopropanes in the presence of a slight excess of diazomethane (2-4 equiv) and a low loading of Pd(OAc)(2) (<0.5 mol %). Diazoethane and diazobutane can also be employed and yield silyl cyclopropanes with diastereoselectivities of up to 10:1 for the trans isomer. When conducted on a 4 g scale, the reaction only required a catalyst loading of 5x10(-3) mol %, which corresponds to a turnover frequency of 40,000 h(-1). Competition experiments revealed that vinyl silanes can be selectively cyclopropanated in the presence of an aliphatic terminal alkene and styrene. The complex [Pd(0) (2)(DVTMS)(3)] (38, DVTMS = divinyltetramethyldisiloxane) proved to be an exceptionally active catalyst for the cyclopropanation reaction, giving complete conversion at -35 degrees C in 1 min. Intermolecular and intramolecular competition experiments with DVTMS (36), both with Pd(OAc)(2) and 38, provided strong evidence for a Pd(0)(alkenyl silane)(3) resting state. Detailed density functional calculations on the reaction pathways for the cyclopropanation of trimethylvinylsilane and DVTMS by diazomethane with Pd(0) corroborated the experimental observations. PMID- 19191238 TI - TiO(2)-modified macroporous silica foams for advanced enrichment of multi phosphorylated peptides. AB - Enriching peptides: Novel TiO(2)-modified macroporous materials (Ti-MOSF, see figure) have been synthesized with high surface area, large pore volume, and functional surfaces that are rich in coordinatively unsaturated Ti(IV) species, which can be applied in the specific extraction of phosphopeptides and which show a preferential capture of multi-phosphorylated peptides with low detection limits and high selectivity. PMID- 19191239 TI - Stoichiometric valence and structural valence--two different sides of the same coin: "bonding power". AB - Recent studies use the term valence to describe two distinct aspects of the phenomenon bonding power of an atom. Measured in valence units, one valence term, the classical chemical valence, has integer values and is derived solely from the composition of a compound. The second one, used mainly by solid-state physicists and crystallographers, has non-integer values. It is determined from structure data, which are derived from diffraction experiments, spectroscopy, or quantum chemical calculations. To distinguish clearly between these two types of valencies, the descriptive terms stoichiometric valence and structural valence and the respective symbols (stoich)V and (struct)V should be used. For the majority of crystalline structures, values of (stoich)V and (struct)V, both measured in valence units, differ by less than 5%. However, for p-block atoms with one lone electron pair, differences between (stoich)V and (struct)V of up to 30% have been reported. PMID- 19191241 TI - Topology of electron charge density for chemical bonds from valence bond theory: a probe of bonding types. AB - To characterize the nature of bonding we derive the topological properties of the electron charge density of a variety of bonds based on ab initio valence bond methods. The electron density and its associated Laplacian are partitioned into covalent, ionic, and resonance components in the valence bond spirit. The analysis provides a density-based signature of bonding types and reveals, along with the classical covalent and ionic bonds, the existence of two-electron bonds in which most of the bonding arises from the covalent-ionic resonance energy, so called charge-shift bonds. As expected, the covalent component of the Laplacian at the bond critical point is found to be largely negative for classical covalent bonds. In contrast, for charge-shift bonds, the covalent part of the Laplacian is small or positive, in agreement with the weakly attractive or repulsive character of the covalent interaction in these bonds. On the other hand, the resonance component of the Laplacian is always negative or nearly zero, and it increases in absolute value with the charge-shift character of the bond, in agreement with the decrease of kinetic energy associated with covalent-ionic mixing. A new interpretation of the topology of the total density at the bond critical point is proposed to characterize covalent, ionic, and charge-shift bonding from the density point of view. PMID- 19191240 TI - Enantioselective total synthesis of (-)-candelalides A, B and C: potential Kv1.3 blocking immunosuppressive agents. AB - Novel Kv1.3 blocking immunosuppressants, (-)-candelalides A, B and C, were efficiently synthesized for the first time in a convergent and unified manner starting from (+)-5-methyl-Wieland-Miescher ketone. The synthetic method involved the following key steps: i) a strategic [2,3]-Wittig rearrangement of a stannylmethyl ether to install the stereogenic center at C9 and the exo-methylene function at C8 present in the decalin portion; ii) a straightforward coupling of a trans-decalin portion (BC ring) and a gamma-pyrone moiety through the C16-C3' bond to assemble the requisite carbon framework; and iii) a construction of a characteristic di or tetrahydropyran ring (A ring) by internal nucleophilic ring closure of a hydroxy aldehyde or a hydroxy epoxide. The present total synthesis has fully established the absolute configuration of these natural products. PMID- 19191242 TI - Copper(I) zeolites as heterogeneous and ligand-free catalysts: [3+2] cycloaddition of azomethine imines. AB - Copper(I)-exchanged zeolites were used as heterogeneous ligand-free catalysts for [3+2] cycloaddition of azomethine ylides, which allows versatile, efficient, and highly regioselective synthesis of pyrazolone derivatives. These cheap and easy to-prepare catalysts exhibit wide scope and compatibility with functional groups. They are very simple to use, easy to remove (by simple filtration), and recyclable (up to six times without loss of activity). PMID- 19191244 TI - Optical response sensitive to the assembly in a molecular material: ultrathin film with a vanishing electronic absorption. AB - Optical properties of single-component molecular materials can often be described in terms of modest perturbations on the responses of the molecular building blocks; in specific systems, the modifications may be more pronounced. In all cases, however, the effects are broadly independent of the form of assembly, such as thin films, nanostructures and crystals, even though variations may exist at the quantitative level. We present a very different situation encountered with an amphiphilic molecule possessing a strongly polar head group that shows similar electronic absorption spectra in the solution as well as the bulk solid state, but a dramatically different one in ultrathin Langmuir-Blodgett (LB) films. The lowest energy absorption is switched off completely in the latter. It is demonstrated further that a metastable form of the LB film exhibiting the normal molecular spectrum can be generated, although its lowest energy absorption decays and vanishes over a period of time. The Langmuir films formed at the air/water interface and the corresponding LB films are examined through pressure-area isotherms, microscopy and spectroscopy. The experimental observations coupled with models constructed through computational investigations allowed us to interpret the unusual optical responses, which are a result of the specific supramolecular 2D assembly in the LB film and the impact of the neighboring molecular dipoles on the intramolecular charge transfer transition. The present study illustrates that selective molecular design and directed assembly can be exploited to elicit dramatic changes in physical responses of molecular materials and realize novel effects in molecular nanostructures. PMID- 19191243 TI - Mechanism of Pd(OAc)2/pyridine catalyst reoxidation by O2: influence of labile monodentate ligands and identification of a biomimetic mechanism for O2 activation. AB - The mechanism of catalyst oxidation by O(2) in Pd-catalyzed aerobic oxidation reactions has been the subject of considerable debate, particularly with respect to the reactivity of Pd(II)-hydride species. Here, we describe the use of unrestricted DFT computational methods to investigate the mechanism of catalyst reoxidation with the Pd(OAc)(2)/pyridine catalyst system, one of the most widely used catalysts. These studies probe four different pathways for the formation of a Pd(II)-hydroperoxide species from the reaction of O(2) from the corresponding Pd(II)-hydride [(py)(n)Pd(II)(H)OAc]: 1) a homolytic pathway involving hydrogen atom abstraction by O(2); 2) AcOH reductive elimination to yield a Pd(0) species that subsequently reacts with O(2); 3) migratory insertion of O(2) into a Pd-H bond; and 4) oxidative addition of O(2) to Pd(II) to yield a Pd(IV)(eta(2) peroxo) species. In contrast to previous studies of reactions between O(2) and Pd hydride species, the reductive-elimination pathway (mechanism 2) is significantly more favorable than any of the other pathways. This outcome is traced to the presence of labile ligands (pyridine) that can readily dissociate from Pd to enable the hydride and acetate ligands to occupy cis-coordination sites. These results strongly support the involvement of Pd(0) as an intermediate in the catalytic cycle. Investigations of the mechanism of the reaction of O(2) with the Pd(0) intermediate revealed a novel, previously unrecognized mechanism that yields a Pd-OOH product without proceeding through the intermediacy of a Pd(II)(eta(2)-peroxo) species. This mechanism resembles pathways commonly observed in biological O(2) activation and suggests that noble-metal and biological oxidation mechanisms may be more similar than previously appreciated. PMID- 19191245 TI - Solid-state enantiopure organic nanocubes formed by self organization of a C3 symmetrical tribenzotriquinacene. AB - Cubic aggregation: The racemic C(3)-symmetrical tribenzotriquinacene 1 was synthesized and found to crystallize in cubic aggregates consisting of eight homochiral molecules. The major driving force for this unique supramolecular aggregation may be attributed to the 24 equivalent sub-van der Waals interactions between the bromine atoms located at the convex surface of the triquinacene cores, rather than to polar interactions between the nitrobenzene units within and between the nanocubes. PMID- 19191246 TI - Photo-induced spin transition of Iron(III) compounds with pi-pi intermolecular interactions. AB - Iron(III) spin-crossover compounds [Fe(pap)(2)]ClO(4) (1), [Fe(pap)(2)]BF(4) (2), [Fe(pap)(2)]PF(6) (3), [Fe(qsal)(2)]NCS (4), and [Fe(qsal)(2)]NCSe (5) (Hpap=2-(2 pyridylmethyleneamino)phenol and Hqsal=2-[(8-quinolinylimino)methyl]phenol) were prepared and their spin-transition properties investigated by magnetic susceptibility and Mossbauer spectroscopy measurements. The iron(III) compounds exhibited spin transition with thermal hysteresis. Single crystals of the iron(III) compounds were obtained as suitable solvent adducts for X-ray analysis, and structures in high-spin (HS) and low-spin (LS) states were revealed. Light induced excited-spin-state trapping (LIESST) effects of the iron(III) compounds were induced by light irradiation at 532 nm for 1-3 and at 800 nm for 4 and 5. The activation energy E(a) and the low-temperature tunneling rate k(HL)(T-->0) of iron(III) LIESST compound 1 were estimated to be 1079 cm(-1) and 2.4x10(-8) s( 1), respectively, by HS-->LS relaxation experiments. The Huang-Rhys factor S of 1 was also estimated to be 50, which was similar to that expected for iron(II) complexes. It is thought that the slow relaxation in iron(III) systems is achieved by the large structural distortion between HS and LS states. Introduction of strong intermolecular interactions, such as pi-pi stacking, can also play an important role in the relaxation behavior, because it can enhance the structural distortion of the LIESST complex. PMID- 19191247 TI - Mesoporous nanofibers from dual structure-directing agents in AAO: Mesostructural control and their catalytic applications. AB - Control yourself! The mesoporous silica nanofibers (MSNFs) from dual structure directing agents were fabricated inside the pores of porous alumina (AAO, see figure) by using vapor phase synthesis. The pore structures could be controlled to form a range of structures from mesocellular foams to cylindrical mesopores with long-range order by adding cationic cosurfactants. PMID- 19191248 TI - Particle-based synthesis of peptide arrays. AB - Lithographic methods allow for the combinatorial synthesis of >50,000 oligonucleotides per cm(2), and this has revolutionized the field of genomics. High-density peptide arrays promise to advance the field of proteomics in a similar way, but currently lag behind. This is mainly due to the monomer-by monomer repeated consecutive coupling of 20 different amino acids associated with lithography, which adds up to an excessive number of coupling cycles. Combinatorial synthesis based on electrically charged solid amino acid particles resolves this problem. A color laser printer or a chip addresses the different charged particles consecutively to a solid support, where, when completed, the whole layer of solid amino acid particles is melted at once. This frees hitherto immobilized amino acids to couple all 20 different amino acids to the support in one single coupling reaction. The method should allow for the translation of entire genomes into sets of overlapping peptides to be used in proteome research. PMID- 19191250 TI - A game of two halves? Incentive incompatibility, starting point bias and the bidding game contingent valuation method. AB - The bidding game (BG) method of contingent valuation is one way to increase the precision of willingness to pay (WTP) estimates relative to the single dichotomous choice approach. However, there is evidence that the method may lead to incentive incompatible responses and be associated with starting point bias. While previous studies in health using BGs test for starting point bias, none have also investigated incentive incompatibility. Using a sample of respondents resident in Burkina Faso, West Africa, this paper examines whether the BG method is associated with both incentive incompatibility and starting point bias. We find evidence for both effects. However, average WTP values remained largely unaffected after accounting for both factors in multivariate analyses. The results suggest that the BG method is an acceptable technique in settings where prices for goods are flexible. PMID- 19191249 TI - Observing proteins as single molecules encapsulated in surface-tethered polymeric nanocontainers. AB - Immobilizing biomolecules provides the advantage of observing them individually for extended time periods, which is impossible to accomplish for freely diffusing molecules in solution. In order to immobilize individual protein molecules, we encapsulated them in polymeric vesicles made of amphiphilic triblock copolymers and tethered the vesicles to a cover slide surface. A major goal of this study is to investigate polymeric vesicles with respect to their suitability for protein folding studies. The fact that polymeric vesicles possess an extreme stability under various chemical conditions is supported by our observation that harsh unfolding conditions do not perturb the structural integrity of the vesicles. Moreover, polymerosomes prove to be permeable to GdnHCl and, thereby, ideally suited for unfolding and refolding studies with encapsulated proteins. We demonstrate this with encapsulated phosphoglycerate kinase, which was fluorescently labeled with Atto655, a dye that exhibits pronounced photoinduced electron transfer (PET) to a nearby tryptophan residue in the native state. Under unfolding conditions, PET was reduced, and we monitored alternating unfolding and refolding conditions for individual encapsulated proteins. PMID- 19191251 TI - Hospital type and patient outcomes: an empirical examination using AMI readmission and mortality records. AB - This paper investigates whether there are differences in patient outcomes across different types of hospitals using patient-level data on readmission and mortality associated with acute myocardial infarction (AMI). Hospitals are grouped according to their ownership type (private, public teaching, public non teaching) and their location (metropolitan, country and remote country). Using data collected from 130 Victorian hospitals on 19,000 patients admitted to a hospital with their first AMI between January 2001 and December 2003, we consider how the likelihood of unplanned re-admission and mortality varies across hospital type. We find that there are significant differences across hospital types in the observed patient outcomes - private hospitals persistently outperform public hospitals. PMID- 19191252 TI - Beneficiary price sensitivity in the Medicare prescription drug plan market. AB - The Medicare stand-alone prescription drug plan (PDP) came into existence in 2006 as part of the Medicare prescription drug benefit. It is the most popular plan type among Medicare drug plans and large numbers of plans are available to all beneficiaries. In this article we present the first analysis of beneficiary price sensitivity in the PDP market. Our estimate of elasticity of enrollment with respect to premium, -1.45, is larger in magnitude than has been found in the Medicare HMO market. This high degree of beneficiary price sensitivity for PDPs is consistent with relatively low product differentiation, low fixed costs of entry in the PDP market, and the fact that, in contrast to changing HMOs, beneficiaries can select a PDP without disrupting doctor-patient relationships. PMID- 19191254 TI - Reliability, validity and responsiveness of a new leisure index: the Patient Specific Leisure Scale (PSLS). AB - OBJECTIVES: To investigate the reliability, validity and responsiveness of a new Patient-Specific Leisure Scale (PSLS), constructed to identify goals and outcomes for individual patients with rheumatoid arthritis (RA). METHODS: Forty-nine patients with RA were used to evaluate test-retest reliability, and 100 consecutive RA patients were used for construct validity. Twenty-five RA patients, commencing with treatment on tumour necrosis factor (TNF) inhibitors, were evaluated before the start and after three months of therapy, to test responsiveness. The most important leisure activity (as judged by the patients) was used when evaluating reliability and validity. The perceived difficulty with each activity was scored from 0 to 10 (0 = able to perform activity without difficulty, 10 = unable to perform activity). RESULTS: Test-retest reliability indicated a good agreement (0.62-0.87) using weighted kappa. Construct validity was demonstrated by modest positive correlation between leisure activity and Health Assessment Questionnaire (HAQ) (r(s) = 0.27, p = 0.005) visual analogue scale (VAS) pain (r(s) = 0.28, p = 0.004) VAS global (r(s) = 0.22, p = 0.027), VAS fatigue (r(s) = 0.24, p = 0.013), joint index of 28 swollen joints (r(s) = 0.22, p = 0.027) and negative correlations with short-form-36 (SF-36) physical functioning (r(s) = -0.18, p = 0.008), bodily pain (r(s) = -0.31, p < 0.001), general health (r(s) = -0.23, p = 0.019), vitality (r(s) = -0.31, p < 0.001), social function (r(s) = -0.24, p = 0.016) and role-emotional (r(s) = -0.28, p = 0.005). Mean improvement for the most important leisure activity was 1.36, (p = 0.036, 95% confidence interval 0.10-2.62). Standardized response mean and effect size for the most important activity in PSLS was 1.05 and 0.72, respectively, and for HAQ 0.34 and 0.28, respectively. CONCLUSIONS: PSLS appears to be feasible, reliable, valid and responsive for measuring leisure activities in RA. It provides both an individual result which is useful in clinical work, and results at a group level. PMID- 19191255 TI - Silylation can be omitted in gas-liquid chromatographic/mass spectrometric analysis of ergosterol. PMID- 19191256 TI - Association studies and gene expression analyses of the DISC1-interacting molecules, pericentrin 2 (PCNT2) and DISC1-binding zinc finger protein (DBZ), with schizophrenia and with bipolar disorder. AB - Disrupted-in-Schizophrenia 1 (DISC1) and its molecular cascade have been implicated in the pathophysiology of major psychoses. Previously, we identified pericentrin 2 (PCNT2) and DISC1-binding zinc finger protein (DBZ) as binding partners of DISC1; further, we observed elevated expression of PCNT2 in the postmortem brains and in the lymphocytes of bipolar disorder patients, compared to controls. Here, we examined the association of PCNT2 with schizophrenia in a case-control study of Japanese cohorts. We also examined the association of DBZ with schizophrenia and with bipolar disorder, and compared the mRNA levels of DBZ in the postmortem brains of schizophrenia, bipolar and control samples. DNA from 180 schizophrenia patients 201 controls were used for the association study of PCNT2 and DBZ with schizophrenia. Association of DBZ with bipolar disorder was examined in DNA from 238 bipolar patients and 240 age- and gender-matched controls. We observed significant allelic and genotypic associations of the PCNT2 SNPs, rs2249057, rs2268524, and rs2073380 (Ser/Arg) with schizophrenia; the association of rs2249057 (P = 0.002) withstand multiple testing correction. Several two SNP- and three SNP-haplotypes showed significant associations; the associations of haplotypes involving rs2249057 withstand multiple testing correction. No associations were observed for DBZ with schizophrenia or with bipolar disorder; further, there was no significant difference between the DBZ mRNA levels of control, schizophrenia and bipolar postmortem brains. We suggest a possible role of PCNT2 in the pathogenesis of schizophrenia. Abnormalities of PCNT2, the centrosomal protein essential for microtubule organization, may be suggested to lead to neurodevelopmental abnormalities. PMID- 19191257 TI - Accelerated tryptic digestion of proteins in plasma for absolute quantitation using a protein internal standard by liquid chromatography/tandem mass spectrometry. PMID- 19191258 TI - Re: First vaginal delivery at an older age: does it carry an extra risk for the development of stress urinary incontinence? Groutz A, Helpman L, Gold R, Pauzner D, Lessing JB, Gordon D. 2007. Neurourol Urodyn 26:779-782. PMID- 19191259 TI - Prevalence of urinary incontinence and associated risk factors in nursing home residents: a systematic review. AB - AIMS: To determine not only prevalence rates of urinary incontinence (UI) in nursing home residents but also factors influencing these prevalence rates, and to provide an overview of risk factors associated with UI in this group. METHODS: A systematic review was performed using multiple databases including MEDLINE, EMBASE, CINAHL, PsycINFO and the Cochrane Library from January 1997 to April 2008. In addition, the bibliographies of all relevant articles were searched. Two authors independently assessed the eligibility of all studies and extracted data on study design, population characteristics, definition of incontinence, measurement instrument, risk factors and prevalence rates. RESULTS: Twelve articles containing 16 studies met the eligibility criteria. Prevalence rates of UI in nursing home residents ranged from 43% to 77% (median 58%). When comparing studies, the influencing factors on UI prevalence of age and sex were identified. In total 45 risk factors were described. Within individual study populations, sex, age, cognitive function, dementia, bedfast and locomotion were associated with UI. CONCLUSIONS: UI prevalence rates in nursing homes are high and the influencing factors poorly understood. Although important risk factors similar to those in the general population have been identified, risk factors related to the care process should be further investigated. PMID- 19191260 TI - Waiting times and socioeconomic status among elderly Europeans: evidence from SHARE. AB - Waiting times for specialist consultation and non-emergency surgery are often considered an equitable rationing mechanism in the public healthcare sector, because access to care is not based on socioeconomic status. This study tests empirically this claim using data from the Survey of Health, Ageing and Retirement in Europe (SHARE). The sample includes nine European countries: Austria, Denmark, France, Germany, Greece, Italy, the Netherlands, Spain and Sweden. For specialist consultation, we find that individuals with high education experience a reduction in waiting times of 68% in Spain, 67% in Italy and 34% in France (compared with individuals with low education). Individuals with intermediate education report a waiting-time reduction of 74% in Greece (compared with individuals with low education). There is also evidence of a negative and significant association between education and waiting times for non-emergency surgery in Denmark, the Netherlands and Sweden. High education reduces waits by 66, 32 and 48%, respectively. We also find income effects, although generally modest. An increase in income of 10 000 Euro reduces waiting times for specialist consultation by 8% in Germany and waiting times for non-emergency surgery by 26% in Greece. Surprisingly, an increase in income of 10 000 Euro increases waits by 11% in Sweden. PMID- 19191261 TI - Simultaneous determination of inositol and inositol phosphates in complex biological matrices: quantitative ion-exchange chromatography/tandem mass spectrometry. AB - myo-Inositol (Ins) and myo-inositol phosphates (InsPs) are widely distributed in plants and animals. The evaluation of the distribution of Ins and InsPs in cells and plant sources can impact the understanding of their role in nutrition, cellular processes and diseases, and how they may be modulated by diet. We developed an anion-exchange chromatography/tandem mass spectrometry (HPLC/ESI MS/MS) method for the separation and simultaneous quantitation of Ins and different naturally occurring phosphorylated inositol compounds. Chromatographic separation was achieved in 30 min on a commercial anion-exchange column (0.5 x 150 mm) using a gradient of 200 mM ammonium carbonate buffer (pH 9.0) and 5% methanol in H(2)O. Analytes were identified by selective reaction monitoring using a triple quadrupole mass spectrometer in negative ion electrospray ionization mode. Adenosine 5'-monophosphate was used as a general internal standard for quantitation. Detection is linear in the range of 0.25-400 pmol for Ins, InsP(1), InsP(4), and InsP(5), 40-400 pmol for InsP(2) and InsP(3), and 60 400 pmol for InsP(6), with a minimum r(2) > 0.994. The limit of detection is 0.25 pmol with a signal-to-noise ratio of 10:1 for all analytes. The intra-day and inter-day variations were within 17% at three concentration levels. Recovery values for the seven analytes spiked into extraction solution or different matrices were between 63 and 121%. Using this approach, Ins and InsPs were measured in three different plant samples and in cultured cells, illustrating significant differences in the distribution of inositol compounds in food samples compared to cells and between cell types. PMID- 19191262 TI - Increased genomic instability and altered chromosomal protein phosphorylation timing in HRAS-transformed mouse fibroblasts. AB - The RAS-mitogen-activated protein kinase signaling pathway is often deregulated in cancer cells. In metastatic HRAS-transformed mouse fibroblasts (Ciras-3), the RAS-MAPK pathway is constitutively activated. We show here that Ciras-3 cells exhibit a higher incidence of chromosomal instability than 10T1/2 cells, including higher levels of clonal and nonclonal chromosomal aberrations. Stimulation of serum starved 10T1/2 and Ciras-3 cells with phorbol esters (TPA) results in the phosphorylation of histone H3 at serine 10 and serine 28. Regardless of the increased genomic instability in Ciras-3 cells, TPA-induced H3 phosphorylated at serine 10 and H3 phosphorylated at serine 28 partitioned into distinct nuclear subdomains as they did in the parental cells. However, the timing of the response of the H3 phosphorylation event to TPA induction was delayed in Ciras-3 cells. Further Ciras-3 cells, which have a more open chromatin structure, had increased steady state levels of phosphorylated H3 and HMGN1 relative to parental 10T1/2 cells. TPA-induced H3 phosphorylated at serine 10 and 28 were colocalized with the transcriptionally initiated form of RNA polymerase II in 10T1/2 and Ciras-3 cells. Chromatin immunoprecipitation assays demonstrated that TPA-induced H3 phosphorylation at serine 28 was associated with the immediate early JUN promoter, providing direct evidence that this histone post translational modification is associated with transcriptionally active genes. Together our results demonstrate the increased genomic instability and alterations in the epigenetic program in HRAS-transformed cells. PMID- 19191263 TI - Laser desorption/ionization mass spectrometric study on asphaltenes from different heavy crude oils and products originating from their thermal treatment. PMID- 19191264 TI - Effect of fabrication process parameters on the apex-radius of STM tungsten nanotip. AB - Scanning tunneling microscope (STM) has found a wide application in nanoscience and nanotechnology. This microscope uses an ultra-sharp metallic tip for image acquisition. Resolution of STM images depends largely on the radius of the tip apex; the smaller the radius the higher the resolution. In the present research, an experimental set-up was designed and implemented for fabrication of STM tungsten nanotip using electrochemical-etching method with the aim of optimization of nanotip fabrication process parameters by using Taguchi method. These parameters are electrolyte concentration, immersion length of the tungsten wire, inner diameter of the cathode tube, and the process voltage. It was found that the optimum level of the process parameters for gaining minimum radius of the nanotip apex is electrolyte concentration of 2 M/lit, wire immersion length of 4 mm, cathode tube inner diameter of 55 mm, and voltage of 3.5 V within the range of experiments. By setting the process parameters on the optimum level, the radius of the nanotip apex was decreased by five times in comparison to the mean value of the experimental results. The radius of the nanotip apex was improved down to about 10 nm under the optimum conditions. SCANNING 31: 65-74, 2009. (c) 2009 Wiley Periodicals, Inc. PMID- 19191265 TI - Confocal scanning optical microscopy of a 3-million-year-old Australopithecus afarensis femur. AB - Portable confocal scanning optical microscopy (PCSOM) has been specifically developed for the noncontact and nondestructive imaging of early human fossil hard tissues, which here we describe and apply to a 3-million-year-old femur from the celebrated Ethiopian skeleton, "Lucy," referred to Australopithecus afarensis. We examine two bone tissue parameters that demonstrate the potential of this technology. First, subsurface reflection images from intact bone reveal bone cell spaces, the osteocyte lacunae, whose density is demonstrated to scale negatively with body size, reflecting aspects of metabolism and organismal life history. Second, images of a naturally fractured cross section near to Lucy's femoral mid-shaft, which match in sign those of transmitted circularly polarized light, reveal relative collagen fiber orientation patterns that are an important indicator of femoral biomechanical efficacy. Preliminary results indicate that Lucy was characterized by metabolic constraints typical for a primate her body size and that in her femur she was adapted to habitual bipedalism. Limitations imposed by the transport and invasive histology of unique or rare fossils motivated development of the PCSOM so that specimens may be examined wherever and whenever nondestructive imaging is required. PMID- 19191266 TI - Comprehensive genetic and functional characterization of IPH-926: a novel CDH1 null tumour cell line from human lobular breast cancer. AB - Infiltrating lobular breast cancer (ILBC) is a clinically and biologically distinct tumour entity defined by a characteristic linear cord invasion pattern and inactivation of the CDH1 tumour suppressor gene encoding for E-cadherin. ILBCs also lack beta-catenin expression and show aberrant cytoplasmic localization of the E-cadherin binding protein p120-catenin. The lack of a well characterized ILBC cell line has hampered the functional characterization of ILBC cells in vitro. We report the establishment of a permanent ILBC cell line, named IPH-926, which was derived from a patient with metastatic ILBC. The DNA fingerprint of IPH-926 verified genetic identity with the patient and had no match among the human cell line collections of several international biological resource banks. IPH-926 expressed various epithelial cell markers but lacked expression of E-cadherin due to a previously unreported, homozygous CDH1 241ins4 frameshift mutation. Detection of the same CDH1 241ins4 mutation in archival tumour tissue of the corresponding primary ILBC proved the clonal origin of IPH 926 from this particular tumour. IPH-926 also lacked beta-catenin expression and showed aberrant cytoplasmic localization of p120-catenin. Array-CGH analysis of IPH-926 revealed a profile of genomic imbalances that included many distinct alterations previously observed in primary ILBCs. Spectral karyotyping of IPH-926 showed a hyperdiploid chromosome complement and numerous clonal, structural aberrations. IPH-926 cells were anti-cancer drug-resistant, clonogenic in soft agar, and tumourigenic in SCID mice. In xenograft tumours, IPH-926 cells recapitulated the linear cord invasion pattern that defines ILBCs. In summary, IPH-926 significantly extends the biological spectrum of the established breast cancer cell lines and will facilitate functional analyses of genuine human ILBC cells in vitro and in vivo. PMID- 19191267 TI - Gloss phenomena and image analysis of atomic force microscopy in molecular and cell biology. AB - Proper sample preparation, scan setup, data collection and image analysis are key factors in successful atomic force microscopy (AFM), which can avoid gloss phenomena effectively from unreasonable manipulations or instrumental defaults. Fresh cleaved mica and newly treated glass cover were checked first as the substrates for all of the sample preparation for AFM. Then, crystals contamination from buffer was studied separately or combined with several biologic samples, and the influence of scanner, scan mode and cantilever to data collection was also discussed intensively using molecular and cellular samples. At last, images treatment and analysis with off-line software had been focused on standard and biologic samples, and artificial glosses were highly considered for their high probability. SCANNING 31: 49-58, 2009. (c) 2009 Wiley Periodicals, Inc. PMID- 19191268 TI - How do physician assessments of patient preferences for colorectal cancer screening tests differ from actual preferences? A comparison in Canada and the United States using a stated-choice survey. AB - BACKGROUND: Patient preferences can affect colorectal cancer (CRC) screening test use. We compared utility-based preferences for alternative CRC screening tests from a stated-preference discrete-choice survey of the general population and physicians in Canada and the United States. METHODS: General population respondents (Canada, n = 501; US, n = 1087) participated in a survey with 12 choice scenarios and 9 CRC screening test attributes. Physicians (n = 100, both Canada and US) reported expected patient preferences. We estimated relative importance of attributes using bivariate probit regression analysis and calculated willingness-to-pay for various CRC screening tests. RESULTS: In 28 and 31% of scenarios, Canadian and US respondents, respectively, chose no screening over a hypothetical test. Canadian (45%) and US (46%) physicians expected patients to choose no screening more often. For all groups the most important attribute was sensitivity, but physicians' perception of patients' preferences are significantly different from actual preferences. Other key attributes are those related to test performance or the testing process. Fecal DNA, colonoscopy, and virtual colonoscopy were the most preferred tests by all groups, but respondents were willing-to-pay more than physicians predicted. CONCLUSION: Physicians' perception of patients' preferences are quite different from those of the general population. However, among general population and physicians, Canadian and US preferences were similar. PMID- 19191271 TI - Automatic phase correction of 2D NMR spectra by a whitening method. AB - A new method for the automatic phase correction of multidimensional NMR spectra is described. It is based on the whitening concept formulated as the 'maximization of the number of white pixels into a bitmap that corresponds to the spectrum'. This process of maximization can be factorized along the individual axes of the spectrum and this property makes the method robust and fast. It employs a statistic measure based on a large number of spectral data points and, for this reason, is very tolerant to low signal-to-noise ratio (SNR) and local artifacts. The algorithm can efficiently phase either homonuclear or heteronuclear experiments and, unlike other previous methods, it can also process automatically spectra containing positive or negative peaks so that it is not necessary to deal with individual or special cases. PMID- 19191270 TI - Replication of genetic associations as pseudoreplication due to shared genealogy. AB - The genotypes of individuals in replicate genetic association studies have some level of correlation due to shared descent in the complete pedigree of all living humans. As a result of this genealogical sharing, replicate studies that search for genotype-phenotype associations using linkage disequilibrium between marker loci and disease-susceptibility loci can be considered as "pseudoreplicates" rather than true replicates. We examine the size of the pseudoreplication effect in association studies simulated from evolutionary models of the history of a population, evaluating the excess probability that both of a pair of studies detect a disease association compared to the probability expected under the assumption that the two studies are independent. Each of nine combinations of a demographic model and a penetrance model leads to a detectable pseudoreplication effect, suggesting that the degree of support that can be attributed to a replicated genetic association result is less than that which can be attributed to a replicated result in a context of true independence. PMID- 19191272 TI - Modeling longitudinal count data with dropouts. AB - This paper explores the utility of different approaches for modeling longitudinal count data with dropouts arising from a clinical study for the treatment of actinic keratosis lesions on the face and balding scalp. A feature of these data is that as the disease for subjects on the active arm improves their data show larger dispersion compared with those on the vehicle, exhibiting an over dispersion relative to the Poisson distribution. After fitting the marginal (or population averaged) model using the generalized estimating equation (GEE), we note that inferences from such a model might be biased as dropouts are treatment related. Then, we consider using a weighted GEE (WGEE) where each subject's contribution to the analysis is weighted inversely by the subject's probability of dropout. Based on the model findings, we argue that the WGEE might not address the concerns about the impact of dropouts on the efficacy findings when dropouts are treatment related. As an alternative, we consider likelihood-based inference where random effects are added to the model to allow for heterogeneity across subjects. Finally, we consider a transition model where, unlike the previous approaches that model the log-link function of the mean response, we model the subject's actual lesion counts. This model is an extension of the Poisson autoregressive model of order 1, where the autoregressive parameter is taken to be a function of treatment as well as other covariates to induce different dispersions and correlations for the two treatment arms. We conclude with a discussion about model selection. PMID- 19191273 TI - Efficient solid-phase synthesis of sulfotyrosine peptides using a sulfate protecting-group strategy. AB - Double protection: Efficient Fmoc-based solid-phase synthesis (SPPS) of sulfotyrosine (sY) peptides is achieved by incorporating the sY residue(s) as a dichlorovinyl-protected (DCV) sulfodiester(s) and using 2-methylpiperidine for Fmoc removal. After removal of the other protecting groups, the DCV group could be cleaved by mild hydrogenolysis giving the sY peptides in good yield. PMID- 19191274 TI - Self-assembly of fivefold-symmetric molecules on a threefold-symmetric surface. AB - Buckybowls: The adsorption of penta-tert-butylcorannulene, a molecule with fivefold symmetry, on Cu(111), a surface with threefold symmetry, is investigated by scanning tunneling microscopy complemented by structure calculations. The symmetry mismatch is resolved by the formation of threefold-symmetric subunits consisting of three molecules, which combine with single molecules to form a nearly perfect filling of the plane (see picture). PMID- 19191275 TI - Electrochemical gene-function analysis for single cells with addressable microelectrode/microwell arrays. AB - To each his own: An addressable electrochemical device consisting of orthogonally arranged rows and columns of electrodes has been constructed to monitor protein expression in genetically engineered cells at the single-cell level. The response based on redox cycling reflected the different expression levels of the enzyme from individual HeLa cells transfected with a plasmid vector including secreted alkaline phosphatase. PMID- 19191276 TI - Morphology, binding behavior and MR-properties of paramagnetic collagen-binding liposomes. AB - Collagen is an important component of the extracellular matrix (ECM) and plays an important role in normal tissue maturation and in pathological processes such as atherosclerosis and myocardial infarction. The diagnostics of the latter diseases using MRI could strongly benefit from the use of collagen-specific contrast agents. The current study aimed to develop a bimodal liposomal MR contrast agent that was functionalized with CNA35, a collagen adhesion protein of the Staphylococcus aureus bacterium. The liposomes were characterized in terms of CNA35 protein conjugation and loading. The overall morphology was assessed with DLS and cryo-TEM, while cryo-TEM tomography was used to visualize the protein coverage of the liposomes. The binding properties of the contrast agent were investigated using a fluorescence assay based on the rhodamine content of the liposomes. The bulk relaxivity was determined using regular relaxometry while the MR-properties of liposomes in their bound state were studied using NMR depth profiling. This CNA35 functionalized contrast agent and the set of in vitro experiments we performed indicate the potential of this technology for in vivo molecular imaging of collagen. PMID- 19191277 TI - Evidence for benefit of transversus abdominis training alone or in combination with pelvic floor muscle training to treat female urinary incontinence: A systematic review. AB - AIMS: Pelvic floor muscle training (PFMT) has Level A evidence to treat female urinary incontinence (UI). Recently, indirect training of the pelvic floor muscles (PFM) via the transversus abdominis muscle (TrA) has been suggested as a new method to treat UI. The aim of this article is to discuss whether there is evidence for a synergistic co-contraction between TrA and PFM in women with UI, whether TrA contraction is as effective, or more effective than PFMT in treating UI and whether there is evidence to recommend TrA training as an intervention strategy. METHODS: A computerized search on PubMed, and hand searching in proceedings from the meetings of the World Confederation of Physical Therapy (1993-2007), International Continence Society and International Urogynecology Association (1990-2007) were performed. RESULTS: While a co-contraction of the TrA normally occurs with PFM contraction, there is evidence that a co-contraction of the PFM with TrA contraction can be lost or altered in women with UI. No randomized controlled trials (RCTs) were found comparing TrA training with untreated controls or sham. Two RCTs have shown no additional effect of adding TrA training to PFMT in the treatment of UI. CONCLUSIONS: To date there is insufficient evidence for the use of TrA training instead of or in addition to PFMT for women with UI. Neurourol. Urodyn. 28:368-373, 2009. (c) 2009 Wiley-Liss, Inc. PMID- 19191278 TI - Voiding dysfunction in patients with Parkinson's disease: impact of neurological impairment and clinical parameters. AB - AIMS: We assessed the lower urinary tract symptoms (LUTS) of patients with Parkinson's disease (PD) and their association with different clinical parameters. METHODS: We prospectively evaluated 110 patients (84 men), with a mean age of 61.8 +/- 9.6 years. Mean duration of the disease was 12.3 +/- 7.2 years. Neurological impairment was assessed by the Hoehn-Yahr and the Unified Parkinson Disease Rating scales. LUTS were assessed by the International Continence Society questionnaire. We evaluated the impact of age, PD duration, neurological impairment, gender, and use of anti-Parkinsonian drugs on the voiding function. RESULTS: On multivariate analysis, voiding dysfunction increased with the neurological impairment, but not with patient's age or disease duration. Quality of life (QOL) was affected by the severity of LUTS, and the symptoms with the worst impact were frequency and nocturia. Sixty-three (57.2%) patients were symptomatic. They did not differ with the asymptomatic as to age and disease duration, but had more severe neurological impairment. No impact on LUTS was associated with the use of levodopa, anticholinergics, and dopamine receptor agonists. Men and women were similarly affected by urinary symptoms. CONCLUSIONS: The severity of the neurological disease is the only predictive factor for the occurrence of voiding dysfunction, which affects men and women alike. PMID- 19191279 TI - Patients with painful bladder syndrome have altered response to thermal stimuli and catastrophic reaction to painful experiences. AB - AIMS: To compare cutaneous sensory thresholds, habituation to somatic stimuli, and tendency towards catastrophic reaction to painful stimuli in patients with Painful Bladder Syndrome (PBS) to controls without PBS. METHOD: Thermal and vibratory sensory thresholds were established in 11 PBS patients and 10 controls at C5, T1, T12, and S3 dermatomes. Supra-threshold thermal stimuli were then applied at T12 and S3 for 60 sec while patients periodically rated the intensity of stimuli using a visual analog scale. A Pain Catastrophizing Scale (PCS) questionnaire was also completed by all participants before testing. RESULTS: PBS patients were less sensitive to warm stimuli in the T12 dermatome than asymptomatic controls (thresholds 36.6 +/- 1.10 degrees C vs. 35.3 +/- 1.0 degrees C, P < 0.02) but otherwise had similar thermal and vibratory thresholds. Habituation to supra-threshold stimuli at T12 and S3 dermatomes was more common in controls than PBS subjects (7 (70%) vs. 2 (18%), P < 0.03 and 9 (90%) vs. 3 (27%), P < 0.008, respectively). The PCS score correlated with the duration of PBS symptoms and with thresholds to warm stimuli at T12 dermatome (rho = 0.65, P < 0.03 and rho = 0.5, P < 0.021, respectively). CONCLUSION: Our data suggests that habituation to stimuli may be impaired and that a catastrophic reaction to perceived stimuli may be involved in the sensory experience of PBS patients and facilitate chronic pain. Neurourol. Urodyn. 28:400-404, 2009. (c) 2009 Wiley Liss, Inc. PMID- 19191280 TI - Anxiety among women attending urodynamic investigation: scoping study for a randomized trial of psychometric intervention. PMID- 19191281 TI - Real-time geometric distortion correction for interventional imaging with echo planar imaging (EPI). AB - Many MR-guided interventional procedures rely on fast imaging sequences for providing images in real-time with a precise relation between the target position in the image and its true position. Echo-planar imaging (EPI) methods are very fast but prone to geometric distortions. Here, we propose a correction method designed for real-time conditions, adapting existing approaches based on dual EPI acquisition with varying echo times. The method is demonstrated with MR thermometry for guiding thermal therapies. The proposed approach imposes a small penalty in acquisition speed but adds negligible latency to data processing, an important element for interventions of mobile organs. PMID- 19191282 TI - Flow-induced disturbances in balanced steady-state free precession images: means to reduce or exploit them. AB - In this work computer simulations and phantom measurements are presented that show the effect of flow on in-plane balanced steady-state free precession images. The images were studied for various flow velocities, excitation regions, relaxation times, RF-pulse angles, and off-resonance frequencies. The work shows that flow-induced disturbances are present in the images, but can be reduced by the application of inhomogeneous excitation regions. Also, a velocity quantification method that utilizes the disturbances was developed and proved to quantify flow velocities accurately. The work concluded that the flow-induced disturbances can be reduced to improve image quality, but can also be exploited to quantify the flow velocity. PMID- 19191283 TI - Myelin water measurement in the spinal cord. AB - The desire to monitor the spatial-temporal characteristics of myelination in the spinal cord (SC), in the context of pathological change in demyelinating diseases or proposed neuroregenerative protocols, has led to an interest in noninvasive image-based myelin measurement methods. We present one strategy: a magnetic resonance-based measure that capitalizes on the characteristics of T(2) relaxation of water compartmentalized within tissue. In this study, 32-echo relaxation studies for measuring the myelin water fraction (MWF) were applied in healthy control SC in vivo using a sagittal inversion recovery multiecho sequence, and findings were supported with supplemental studies in bovine SC samples in vitro. Mean human MWF varied according the level of the SC examined: cervical, thoracic, and lumbar MWF was found to be 21.8 (SD=2.1)%, 24.3 (3.6)%, and 11.4 (6.4)%, respectively. Noteworthy reductions were observed in areas consistent with the expected locations of the cervical and lumbar enlargements. Average bovine MWF was 30.0 (2.7)% in white matter and 8.2 (0.4)% in gray matter. The potential applications of T(2) measurement in SC, both in characterizing disease processes like multiple sclerosis and in monitoring neuroregenerative therapies, should encourage future research in this area. PMID- 19191284 TI - Compatible dual-echo arteriovenography (CODEA) using an echo-specific K-space reordering scheme. AB - An improved dual-echo sequence magnetic resonance (MR) imaging technique was developed to simultaneously acquire a time-of-flight MR angiogram (MRA) and a blood oxygenation level-dependent MR venogram (MRV) in a single MR acquisition at 3 T. MRA and MRV require conflicting scan conditions (e.g., excitation RF profile, flip angle, and spatial presaturation pulse) for their optimal image quality. This conflict was not well counterbalanced or reconciled in previous methods reported for simultaneous acquisition of MRA and MRV. In our dual-echo sequence method, an echo-specific K-space reordering scheme was used to uncouple the scan parameter requirements for MRA and MRV. The MRA and MRV vascular contrast was enhanced by maximally separating the K-space center regions acquired for the MRA and MRV, and by adjusting and applying scan parameters compatible between the MRA and MRV. As a preliminary result, we were able to acquire a simultaneous dual-echo MRA and MRV with image quality comparable to that of the conventional single-echo MRA and MRV that were acquired separately at two different sessions. Furthermore, integrated with tilted optimized nonsaturating excitation and multiple overlapping thin-slab acquisition techniques, our dual echo MRA and MRV provided seamless vascular continuity over a large coverage volume of the brain anatomy. PMID- 19191286 TI - Effects of echo time on diffusion quantification of brain white matter at 1.5 T and 3.0 T. AB - The aim was to investigate the effects of echo time (TE) on diffusion quantification of brain white matter. Seven rhesus monkeys (all males; age, 4-6 years; weight, 5-7 kg) underwent diffusion tensor imaging (DTI) with a series of TEs in 1.5 T and 3.0 T MR scanners. The mean diffusivity (MD), fractional anisotropy (FA), primary (lambda(1)), and transverse eigenvalues (lambda(23)) were measured in a region of interest at the bilateral internal capsule. Pearson correlation showed that the FA and lambda(1) increased and lambda(23) decreased with TE both at 1.5 T and 3.0 T except for the MD. Repeated measurement analysis of variance (ANOVA) also showed significantly higher FA and lower MD and lambda(23) at 3.0 T than those at 1.5 T (P<0.01), but no statistical differences were found in lambda(1) between these two field strengths (P=0.709). These findings implied that TE and field strength might influence diffusion quantification in brain white matter. PMID- 19191285 TI - Ultrashort T2* relaxometry for quantitation of highly concentrated superparamagnetic iron oxide (SPIO) nanoparticle labeled cells. AB - A new method was developed to measure ultrashort T(2)* relaxation in tissues containing a focal area of superparamagnetic iron oxide (SPIO) nanoparticle labeled cells in which the T(2)* decay is too short to be accurately measured using regular gradient echo T(2)* mapping. The proposed method utilizes the relatively long T(2) relaxation of SPIO-labeled cells and acquires a series of spin echo images with the readout echo shifted to sample the T(2)* decay curve. MRI experiments in phantoms and rats with SPIO-labeled tumors demonstrated that it can detect ultrashort T(2)* down to 1 ms or less. The measured T(2)* values were about 10% higher than those from the ultrashort TE (UTE) technique. The shorter the TE, the less the measurements deviated from the UTE T(2)* mapping. Combined with the regular T(2)* mapping, this technique is expected to provide quantitation of highly concentrated iron-labeled cells from direct cell transplantation. PMID- 19191287 TI - Cerebrospinal fluid and interstitial fluid volume measurements in the human brain at 3T with EPI. AB - Cerebrospinal fluid (CSF) diffusion into periventricular white matter is one of the pathophysiological features of hydrocephalus of any kind. In standard clinical routine periventricular hyperintensities, size of the ventricular system, and invasive CSF pressure measurement are the key diagnostic methods. Recently a method called quantitative blood oxygenation level-dependent (qBOLD) was proposed by He and Yablonskiy (Magn Reson Med 2007;57:115-126), which uses the signal evolution of a GESSE sequence to extract quantitative information about hemodynamic parameters. In this study a similar method was used to extract volume fraction information of CSF and interstitial fluid (ISF) in the human brain. A standard gradient recalled echo / echo planar imaging (GRE-EPI) sequence with variable echo time was used for the acquisition of the MRI signal. A first test on healthy subjects yielded consistent results for ISF/CSF volume fraction, which were in good agreement with results found in the literature. PMID- 19191288 TI - HPV detection rate in discordant Pap tests between cytotechnologists and pathologists. AB - When a Pap test is referred for pathologist review, it is accompanied by the cytotechnologist's provisional interpretation. Discordant interpretations between pathologists and cytotechnologists are sometimes noted. The objective is to correlate HPV detection rate with both estimated false-negative fraction (EFNF) and ASC to SIL ratio among discordant ASC cases. ThinPrep Pap tests in which the cytotechnologists' provisional interpretations were up- or down-graded by pathologists to ASC were retrieved between January and December 2006. HPV DNA testing was performed using hybrid capture technique. EFNFs and ASC to SIL ratio were estimated for cytotechnologists and pathologists, respectively. Overall, the EFNF ratio was 3.4% and the high-risk HPV DNA detection rates in cases that were "over-" or "under-" interpreted by technologists were 0.71 and 0.40, respectively. The overall ASC to SIL ratio was 1.41 and the high-risk HPV DNA detection rates in cases that were upgraded or downgraded to ASC were 0.40 and 0.71, respectively. In conclusion, our ASC to SIL ratios and EFNF were within acceptable range. We did not observe any association between ASC to SIL ratio and HPV detection rate in cases that were upgraded or downgraded to ASC by pathologists or between EFNF ratio and HPV detection rate in cases that were "over-interpreted" (or "under-interpreted") by cytotechnologists. The HPV detection rates for ASC cases that were originally "over-interpreted" by cytotechnologists were comparable to the HPV detection rates for LSIL, whereas those that were underinterpreted were comparable to the HPV detection rate for ASC. PMID- 19191289 TI - Cytologic diagnosis of osseous lesions: a review with emphasis on the diagnosis of primary neoplasms of bone. AB - Fine-needle aspiration has been utilized as the initial diagnostic technique at a large number of body sites for over three quarters of a century. As early as the 1930s, fine-needle aspiration (FNA) was used to investigate lesions of the musculoskeletal system. In many early reports, FNA was most frequently and successfully used for the diagnosis of metastatic disease to bone. Less emphasis was placed on its utility for the investigation of primary neoplasms of bone and soft tissue. Current utilization of FNA continues to de-emphasize its application to the diagnosis of primary lesions of the musculoskeletal system. Recent advances in imaging techniques, immunohistochemistry, and molecular diagnostics along with an increasing familiarity among pathologists with the cytologic appearance of primary osseous tumors has led to reevaluation of the technique for investigation of these tumors. The diagnostic accuracy of FNA along with its relatively low cost and high degree of safety makes it a desirable technique for the investigation of primary lesions of the musculoskeletal system. This article reviews issues of diagnostic accuracy, optimal practice procedures, and benefits of the technique including cost reduction. The article will review criteria for selection of appropriate tissue targets for FNA to reduce the number of unsatisfactory specimens. Cytomorphologic features of the more common primary neoplasms of bone will be summarized along with recommendations for the utilization of immunohistochemistry and molecular diagnostics in the work-up of primary neoplasms of bone. PMID- 19191291 TI - Use of streamed internet video for cytology training and education: www.PathLab.org. AB - An Internet-based method is described for submission of video clips to a website editor to be reviewed, edited, and then uploaded onto a video server, with a hypertext link to a website. The information on the webpages is searchable via the website sitemap on Internet search engines. A survey of video users who accessed a single 59-minute FNA cytology training cytology video via the website showed a mean score for usefulness for specialists/consultants of 3.75, range 1 5, n = 16, usefulness for trainees mean score was 4.4, range 3-5, n = 12, with a mean score for visual and sound quality of 3.9, range 2-5, n = 16. Fifteen out of 17 respondents thought that posting video training material on the Internet was a good idea, and 9 of 17 respondents would also consider submitting training videos to a similar website. This brief exercise has shown that there is value in posting educational or training video content on the Internet and that the use of streamed video accessed via the Internet will be of increasing importance. PMID- 19191290 TI - Fine-needle aspiration of breast carcinoma metastatic to follicular variant of papillary thyroid carcinoma. PMID- 19191292 TI - Differentiating between endocervical glandular neoplasia and high grade squamous intraepithelial lesions in endocervical crypts: cytological features in ThinPrep and SurePath cervical cytology samples. AB - A recent audit at our institution revealed a higher number of cases diagnosed as endocervical glandular neoplasia on ThinPrep (TP) cervical cytology samples (9 cases) as opposed to SurePath (SP) (1 case), which on histology showed only high grade cervical intraepithelial neoplasia (CIN) with endocervical crypt involvement (CI). We attempted to ascertain the reasons for this finding by reviewing the available slides of these cases, as well as slides of cases diagnosed as glandular neoplasia on cytology and histology; cases diagnosed as high-grade squamous intraepithelial lesions (HSIL) on cytology which had CIN with CI on histology and cases with mixed glandular and squamous abnormalities diagnosed both cytologically and histologically. Single neoplastic glandular cells and short pseudostratified strips were more prevalent in SP than TP with the cell clusters in glandular neoplasia 3-4 cells thick, in contrast to the dense crowded centre of cell groups in HSIL with CI. The cells at the periphery of groups can be misleading. Cases with HSIL and glandular neoplasia have a combination of the features of each entity in isolation. The diagnosis of glandular neoplasia remains challenging and conversion from conventional to liquid based cervical cytology requires a period of learning and adaptation, which can be facilitated by local audit and review of the cytology slides in cases with a cytology-histology mismatch. PMID- 19191293 TI - Fine-needle cytology of metastatic endometrioid neoplasms: experience with eight cases. AB - Despite the frequency of endometrioid malignancies, few articles in the literature are found concerning their cytopathologic presentation on fine-needle cytology samples. This report describes the cytomorphological findings in eight cases of recurrent or metastatic endometrioid neoplasms on fine-needle cytology samples obtained from various body sites. The cytological findings in metastatic or recurrent endometrioid carcinomas could be classified into five main patterns (i.e.: endometrioid, adeno-squamous, villo-glandular, clear cell, and papillary serous), in analogy to histology. It is the authors' feeling that an adequate knowledge of the cytopathological features of this group of neoplasms may be important in favoring an early detection of their relapses or metastases and may contribute to save diagnostic time and more invasive procedures to the patients. PMID- 19191294 TI - Differentiating reactive mesothelial cells from metastatic adenocarcinoma in serous effusions: the utility of immunocytochemical panel in the differential diagnosis. AB - Differentiating reactive mesothelial cells (RMs) from metastatic adenocarcinoma cells (MAC) in serous fluids based on cytomorphologic features alone can be very challenging. Various immunocytochemical (ICC) markers have been used to maximize the diagnostic accuracy, however, cytopathologists still encounter difficulties in effusion cytologic diagnosis. The aim of this study was to evaluate previous and recent ICC stains to identify the most sensitive and specific markers and the best panel for differentiating RM from MAC. Cell block sections from 41 MAC and 43 RM effusions cases were subjected to ICC staining for MOC-31, BerEp4, carcinoembryonic antigen (CEA), calretinin, HBME-1, CK5/6, and D2-40. For the MAC cases, the sensitivity of BerEp4, MOC-31, and CEA was 82.9, 92.6, and 17%, respectively, and the specificity was 95.3, 93, and 100%, respectively. For the RM cases, the sensitivity of calretinin, CK5/6, D2-40, and HBME-1 was 95.3, 27.9, 58.1, and 93%, respectively, and the specificity was 70.7, 73.1, 75.6, and 82.9%, respectively. The results show that BerEp4 and MOC-31 are highly sensitive and specific for detecting MAC, whereas calretinin and HBME1 are highly sensitive but only modestly specific for detecting RM cases (P < 0.05). Forced entry logistic regression revealed that using MOC-31, BerEp4, HBME-1, and calretinin, is an excellent panel for making correct diagnosis with 97.6% sensitivity in detecting MAC and 90.7% specificity in detecting RM. We conclude that adding a panel of MOC 31, BerEp4, calretinin, and HBME-1 immunostains to routine cytomorphologic features can greatly enhance the diagnostic accuracy of serous effusions. PMID- 19191295 TI - Cervical intraepithelial neoplasia and aneusomy of TERC: assessment of liquid based cytological preparations. AB - The implementation of effective screening programs has decreased the incidence and mortality of cervical carcinoma. However, single screening tests are subjective and carry a significant false-negative rate. Therefore, supplementary tests to support the Papanicolaou (PAP) smear are being developed. Human papillomavirus (HPV) testing has increased the specificity of the PAP smear, but has high sensitivity rate. This has proven unhelpful in low-grade lesions and in young women. Cervical carcinogenesis is a multifactorial disorder. In addition to exposure to oncogenic HPV, which is regarded as the initiator, there must be a promoter to eventuate in invasive disease. The promoter factor appears to be the acquisition of extra copies of chromosome 3q and has been shown to be a constant recurrence in cervical carcinoma (squamous and adenocarcinomas). The 3q region contains the RNA sequence of the human telomerase gene TERC. Recent studies have shown a strong correlation between high-grade cervical lesions and abnormalities of TERC. This study supports the previous studies and examines the status of the TERC gene in low and high-grade cervical intraepithelial lesions, diagnosed on cytology. ASC-H smears are also examined in an attempt to categorize lesions that are more likely to progress. Potentially this may help identify women in need of close clinical follow-up and early treatment. PMID- 19191296 TI - "String of pearls" appearance of blue blobs in postmenopausal atrophy on ThinPrep Pap test. PMID- 19191297 TI - Cytologic diagnosis of pulmonary adenocarcinoma with micropapillary pattern: does it correlate with the histologic findings? AB - Micropapillary adenocarcinoma is associated with poor-prognosis in several organs including the lung. The presence of small tight balls of neoplastic cells devoid of fibrovascular core in cytological preparations (micropapillary tufts) has been described as characteristic of micropapillary adenocarcinoma. In the lung, however, this criterion has not been validated. The cytological material of 46 cases of histologically proven pulmonary adenocarcinoma with a micropapillary component was compared to 33 cases with no micropapillary component to determine the specificity of micropapillary tufts for the histologic diagnosis of micropapillary adenocarcinoma. Other histologic patterns of invasive pulmonary adenocarcinomas (acinar, papillary, and solid) were also compared with patterns of neoplastic cell aggregates in cytological preparations. There were no significant differences in the distribution of micropapillary clusters between the two groups. The positive predictive value for the cytologic diagnosis of a micropapillary component in lung adenocarcinomas was of 64%. Similar findings were observed for other invasive patterns. Therefore, the detection of micropapillary tufts in cytology is not specific for the diagnosis of a pulmonary micropapillary adenocarcinoma in the lung. PMID- 19191298 TI - Usefulness of lavage cytology during endoscopic transpapillary catheterization into the gallbladder in the cytological diagnosis of gallbladder disease. AB - Many studies have reported methods of cell collection involving percutaneous transhepatic cholangiodrainage (PTCD) and fine-needle aspiration cytology for the diagnosis of gallbladder disease. However, few studies have described the use of a transpapillary approach, i.e., endoscopic transpapillary catheterization into the gallbladder (ETCG). In this study, we analyzed cells collected by ETCG to evaluate its usefulness in the cytological diagnosis of gallbladder disease. The subjects were 19 patients who had undergone ETCG for the diagnosis of gallbladder disease. Of these patients, 11 and 8 had gallbladder cancer and benign gallbladder disease, respectively. We also evaluated the diagnostic accuracy of PTCD cytology performed in 15 patients with gallbladder cancer.Specimens were cytologically diagnosed as normal or benign, indeterminate, suspected malignancy, malignant, and inadequate in 47% (9/19), 11% (2/19), 0% (0/19), 37% (7/19), and 5% (1/19) of patients, respectively. Specimens were diagnosed as malignant, indeterminate, normal or benign, and inadequate in 7, 2, 1, and 1, respectively, of the 11 patients diagnosed with gallbladder cancer. The sensitivity and specificity of ETCG cytology were 78 and 100%, respectively, whereas the diagnostic accuracy of PTCD cytology was 20% (3/15). None of the patients developed complications of ETCG. Despite its technical difficulty, ETCG for bile cytology allows the collection of adequate cell numbers from patients with benign disease or gallbladder cancer and facilitates a cytological diagnosis, making it a useful method for collecting cells. PMID- 19191299 TI - Pseudomonas aeruginosa biofilms in a bronchoalveolar lavage. PMID- 19191300 TI - Fine-needle aspiration of extranodal Hodgkin's lymphoma misinterpreted as carcinoma. PMID- 19191301 TI - Mutations and polymorphisms in the proprotein convertase subtilisin kexin 9 (PCSK9) gene in cholesterol metabolism and disease. AB - Hypercholesterolemia is one of the major causes of coronary heart disease (CHD). The genes encoding the low-density lipoprotein receptor and its ligand apolipoprotein B, have been the two genes classically implicated in autosomal dominant hypercholesterolemia (ADH). Our discovery in 2003 of the first mutations of the proprotein convertase subtilisin kexin 9 gene (PCSK9) causing ADH shed light on an unknown actor in cholesterol metabolism that since then has been extensively investigated. Several PCSK9 variants have been identified, some of them are gain-of-function mutations causing hypercholesterolemia by a reduction of low-density lipoprotein (LDL) receptor levels; while others are loss-of function variants associated with a reduction of LDL-cholesterol (LDL-C) levels and a decreased risk of CHD. In this review, we focus on reported variants, and their biological, clinical, and functional relevance. We also highlight the spectrum of hypercholesterolemia or hypobetalipoproteinemia phenotypes that are already associated with mutations in PCSK9. Finally, we present future prospects concerning this therapeutic target that might constitute a new approach to reduce cholesterol levels and CHD, and enhance the effectiveness of other lipid-lowering drugs. PMID- 19191303 TI - Updates on amyotrophic lateral sclerosis: improving patient care. PMID- 19191304 TI - Current hypotheses for the underlying biology of amyotrophic lateral sclerosis. AB - The mechanisms involved in selective motor neuron degeneration in amyotrophic lateral sclerosis remain unknown more than 135 years after the disease was first described. Although most cases have no known cause, mutations in the gene encoding Cu/Zn superoxide dismutase (SOD1) have been implicated in a fraction of familial cases of the disease. Transgenic mouse models with mutations in the SOD1 gene and other ALS genes develop pathology reminiscent of the disorder, including progressive death of motor neurons, and have provided insight into the pathogenesis of the disease but have consistently failed to predict therapeutic efficacy in humans. However, emerging research has demonstrated that mutations and pathology associated with the TDP-43 gene and protein may be more common than SOD1 mutations in familial and sporadic ALS. Putative mechanisms of toxicity targeting motor neurons include oxidative damage, accumulation of intracellular aggregates, mitochondrial dysfunction, defects in axonal transport, growth factor deficiency, aberrant RNA metabolism, glial cell pathology, and glutamate excitotoxicity. Convergence of these pathways is likely to mediate disease onset and progression. PMID- 19191305 TI - Racing against the clock: recognizing, differentiating, diagnosing, and referring the amyotrophic lateral sclerosis patient. AB - Recognition of the early symptoms and signs in amyotrophic lateral sclerosis, exclusion of alternative diagnoses, and referral to a tertiary center can have a significant positive impact on the lives of patients and their caregivers. This article provides the most current amyotrophic lateral sclerosis criteria, as well as helpful clinical clues to the diagnosis. An approach to laboratory testing, electrodiagnostic testing, and imaging to exclude diseases that mimic ALS also are discussed, as are atypical presentations that can confound timely diagnosis. PMID- 19191306 TI - Managing amyotrophic lateral sclerosis: slowing disease progression and improving patient quality of life. AB - It is now possible to slow the disease progression of amyotrophic lateral sclerosis (ALS), but documented improvement in the quality of life of ALS patients has been difficult to quantitate. Putative mechanisms involved in motor neuron degeneration in ALS include oxidative damage, mitochondrial dysfunction, neuroinflammation, growth factor deficiency, and glutamate excitotoxicity. Several pharmacological agents that target these potential targets have demonstrated therapeutic potential in animal models with mutations in the gene encoding Cu/Zn superoxide dismutase (SOD1). Many treatments that have been moderately effective in this animal model have not been successfully translated into effective treatments for humans with ALS. Only the glutamate modulator riluzole has demonstrated efficacy in clinical trials and is approved for treating ALS. Combination treatments may represent a potential therapeutic strategy to more robustly prolong life and preserve function, but only vitamin E with riluzole has been formally studied in clinical trials, and to date, no combination treatments have been found to be more effective than currently available single agents. PMID- 19191307 TI - Outcomes research in amyotrophic lateral sclerosis: lessons learned from the amyotrophic lateral sclerosis clinical assessment, research, and education database. AB - OBJECTIVE: To examine the care of patients with ALS following the publication of the standardized recommendations for the management of patients with amyotrophic lateral sclerosis (ALS) published in 1999 by the American Academy of Neurology. METHODS: Specific aspects of ALS patient management have been evaluated serially using a national Amyotrophic Lateral Sclerosis Clinical Assessment, Research, and Education (ALS CARE) database to encourage compliance with these recommendations and to assure continuing quality improvement. RESULTS: The most recent analysis of 5,600 patients shows interesting epidemiological observations and treatment trends. Proper management of many ALS symptoms has increased substantially since the first publication of the guidelines, and awareness of pseudobulbar affect has increased. Other recommendations are underutilized: Only 9% undergo percutaneous endoscopic gastrostomy, although this procedure was recommended in 22% of patients; and noninvasive positive pressure ventilation was used by only 21% of patients despite being associated with improved 5-year survival rates. INTERPRETATION: This observational database has been a useful tool in monitoring compliance with the standard of care for patients with ALS and may have resulted in greater adherence to guidelines. PMID- 19191311 TI - Initial osteoblast functions on Ti-5Zr-3Sn-5Mo-15Nb titanium alloy surfaces modified by microarc oxidation. AB - This study is intended to evaluate the effects of microarc oxidation (MAO) on the biocompatibility of near beta titanium alloy Ti-5Zr-3Sn-5Mo-15Nb (TLM) in vitro. Two porous bioactive surfaces with different surface characteristics were grown on TLM substrates via MAO process at two different final voltages. Both calcium and phosphorus were incorporated into the oxidized surfaces, and their content was dependent on the voltage applied. Surface roughness was enhanced on the MAO surfaces, which was higher when a higher voltage was applied. After MAO treatment, water contact angles became smaller and surface energies were increased, especially the polar components, which were also related to the MAO final voltage. Cell culture experiments showed an enhanced osteoblasts adhesion, spread, and viability on the microarc oxidized surfaces, and better cell spread and viability were found on the surface formed at 450 V than that at 300 V. No obvious variations in gene expression of integrin beta1 (Itg beta 1), core binding factor-alpha1, osteopontin, collagen type I alpha2-chain, and fibronectin by osteoblasts were observed on different surfaces. The expression of osteocalcin was strikingly increased on MAO surfaces after 72 h, thus indicating enhanced osteoblasts differentiation on MAO surfaces. Interestingly, obvious enhanced bone morphogenetic proteins (BMP)-2 and BMP-4 expression was observed on MAO surfaces, which may be a reason for the enhanced osteoblasts functions on MAO-modified TLM surfaces. PMID- 19191310 TI - Influence of naturally-occurring 5'-pyrophosphate-linked substituents on the binding of adenylic inhibitors to ribonuclease a: an X-ray crystallographic study. AB - Ribonuclease A is the archetype of a functionally diverse superfamily of vertebrate-specific ribonucleases. Inhibitors of its action have potential use in the elucidation of the in vivo roles of these enzymes and in the treatment of pathologies associated therewith. Derivatives of adenosine 5'-pyrophosphate are the most potent nucleotide-based inhibitors known. Here, we use X-ray crystallography to visualize the binding of four naturally-occurring derivatives that contain 5'-pyrophosphate-linked extensions. 5'-ATP binds with the adenine occupying the B(2) subsite in the manner of an RNA substrate but with the gamma phosphate at the P(1) subsite. Diadenosine triphosphate (Ap(3)A) binds with the adenine in syn conformation, the beta-phosphate as the principal P(1) subsite ligand and without order beyond the gamma-phosphate. NADPH and NADP(+) bind with the adenine stacked against an alternative rotamer of His119, the 2'-phosphate at the P(1) subsite, and without order beyond the 5'-alpha-phosphate. We also present the structure of the complex formed with pyrophosphate ion. The structural data enable existing kinetic data on the binding of these compounds to a variety of ribonucleases to be rationalized and suggest that as the complexity of the 5'-linked extension increases, the need to avoid unfavorable contacts places limitations on the number of possible binding modes. PMID- 19191312 TI - The influence of heat treatment and plastic deformation on the bio-degradation of a Mg-Y-RE alloy. AB - In this study the bio-degradation behavior of a Mg-Y-RE alloy in different heat treatment states with respect to the alloy's potential application as biodegradable implant material was investigated by electrochemical impedance spectroscopy in two body-similar fluids. The heat treatments increase the degradation resistance of the alloy and lead to the formation of a thermal oxide layer on the sample surface and to a change in microstructure such as the distribution of yttrium. The varying Y distribution in the alloy does not significantly influence the degradation behavior, and all samples show a similar low polarization resistance. However, samples with a thermal oxide layer, which consists mainly of Y(2)O(3), degrade much more slowly and feature remarkably high polarization resistance. Nevertheless, in some cases localized corrosion attack occurs and drastically impairs performance. Cracks in the oxide layer, intentionally induced by straining of the samples and which in practice could originate from the implantation process, reduce the corrosion resistance. However, these samples perform still better than polished specimens and show a macroscopically homogeneous degradation behavior without localized corrosion. Microscopically, corrosion attacks start at the cracks and undermining of the oxide layer occurs with time. For all the material conditions a remarkable dependence of the degradation rate on the electrolyte is noted. PMID- 19191313 TI - The effects of monoacrylated poly(ethylene glycol) on the properties of poly(ethylene glycol) diacrylate hydrogels used for tissue engineering. AB - This study investigated the effects of poly(ethylene glycol) monoacrylate (PEGMA) on the properties of poly(ethylene glycol) diacrylate (PEGDA)-co-PEGMA hydrogel networks. The PEGMA materials utilized were similar to ligand-linked materials typically copolymerized with PEGDA for use as tissue engineering scaffolds. PEGDA (5-20% wt/wt, 6 kDa) and PEGMA (0-20% wt/wt, 0-43 mM, 5 kDa) were copolymerized by photo-initiated free radical polymerization and the mass swelling ratio and shear modulus of the resulting hydrogels were determined. Increasing the prepolymerization concentration of PEGMA decreased the swelling ratio by up to 42 +/- 1.6% and increased the shear modulus by up to 167 +/- 29.3%, suggesting that PEGMA enhanced gel cross-linking. Analysis of the effective number of cross linked chains per PEGDA, calculated independently from swelling and mechanical data, indicated each PEGDA participated in more cross-links as PEGMA was added. The results suggest that PEGMA-co-PEGDA gels can be formed with higher concentrations of PEGMA-tethered ligands than previously reported allowing the formation of scaffolds with a rich diversity of biological functionalities without sacrificing the integrity of the gel network. PMID- 19191314 TI - A new route to produce starch-based fiber mesh scaffolds by wet spinning and subsequent surface modification as a way to improve cell attachment and proliferation. AB - This study proposes a new route for producing fiber mesh scaffolds from a starch polycaprolactone (SPCL) blend. It was demonstrated that the scaffolds with 77% porosity could be obtained by a simple wet-spinning technique based on solution/precipitation of a polymeric blend. To enhance the cell attachment and proliferation, Ar plasma treatment was applied to the scaffolds. It was observed that the surface morphology and chemical composition were significantly changed because of the etching and functionalization of the fiber surfaces. XPS analyses showed an increase of the oxygen content of the fiber surfaces after plasma treatment (untreated scaffolds O/C:0.32 and plasma-treated scaffolds O/C:0.41). Both untreated and treated scaffolds were examined using a SaOs-2 human osteoblast-like cell line during 2 weeks of culture. The cell seeded on wet-spun SPCL fiber mesh scaffolds showed high viability and alkaline phosphatase enzyme activity, with those values being even higher for the cells seeded on the plasma treated scaffolds. PMID- 19191315 TI - Preparation and characterization of polyethylenimine-coated Fe3O4-MCM-48 nanocomposite particles as a novel agent for magnet-assisted transfection. AB - A new type of magnetic nanoparticle was synthesized using mesoporous silica MCM 48 as a template. Magnetite (Fe(3)O(4)) nanocrystals were incorporated onto the MCM-48 silica structure by thermal decomposition of iron(III) acetylacetonate. The particle size of these Fe(3)O(4)-MCM-48 composite particles is around 300 nm with an iron oxide content of ca. 20% w/w. Measurements from SQUID magnetometry suggest that these nanoparticles possess superparamagnetic properties similar to those of Fe(3)O(4) nanoparticles. By coating positively charged polyethylenimine on to the surface, DNA can be bound onto the Fe(3)O(4)-MCM-48 nanoparticles. Transfection studies showed that these PEI-Fe(3)O(4)-MCM-48 particles were highly effective as a transfection reagent, and a 400% increase of transfection efficiency compared with the commercial products was recorded. PMID- 19191316 TI - In vivo bone biocompatibility and degradation of porous fumarate-based polymer/alumoxane nanocomposites for bone tissue engineering. AB - The objective of this study was to determine how the incorporation of surface modified alumoxane nanoparticles into a biodegradable fumarate-based polymer affects in vivo bone biocompatibility (characterized by direct bone contact and bone ingrowth) and in vivo degradability. Porous scaffolds were fabricated from four materials: poly(propylene fumarate)/propylene fumarate-diacrylate (PPF/PF DA) polymer alone; a macrocomposite consisting of PPF/PF-DA polymer with boehmite microparticles; a nanocomposite composed of PPF/PF-DA polymer and mechanically reinforcing surface-modified alumoxane nanoparticles; and a low-molecular weight PPF polymer alone (tested as a degradation control). Scaffolds were implanted in the lateral femoral condyle of adult goats for 12 weeks and evaluated by micro computed tomography and histological analysis. For all material groups, small amounts of bone, some soft tissue, and a few inflammatory elements were observed within the pores of scaffolds, though many pores remained empty or filled with fluid only. Direct contact between scaffolds and surrounding bone tissue was also observed in all scaffold types, though less commonly. Minimal in vivo degradation occurred during the 12 weeks of implantation in all materials except the degradation control. These results demonstrate that the incorporation of alumoxane nanoparticles into porous PPF/PF-DA scaffolds does not significantly alter in vivo bone biocompatibility or degradation. PMID- 19191317 TI - Intradiscal drug delivery system for the treatment of low back pain. AB - Possible solution to the long-term control of the low back pain (LBP) would be by using an injectable pain drug carrier that can be delivered locally. The drug can be released in a controlled manner. It is also allowed to inject repeatedly more drugs percutaneously with a minimal invasion. The main objective of this study was to develop such a drug delivery system (DDS) for long-term control of discogenic LBP. The DDS consists of in situ forming hydrogel matrix (Pluronic F127 plus sodium hyaluronate) containing microspheres (MS). The solid MS were used for long-term release of the drugs. Both hydrogel matrix and MS contained a model drug, bupivacaine base (BB). The phase transition (liquid at room temperature, gel at around body temperature) could be manipulated by changing the composition of the hydrogel. In vitro test showed that approximately 3% (w/w) of the BB loaded to MS were released during 42 days, demonstrating a good potential for sustained release of bupivacaine. PMID- 19191318 TI - Farnesol-modified biodegradable polyurethanes for cartilage tissue engineering. AB - A bifunctionalized 3,7,11-trimethyl-2,6,10-dodecatrien-1-diaminobutane amide (isoprenoid) was obtained from 3,7,11-trimethyl-2,6,10-dodecatrien-1-ol (farnesol) in a three-step synthesis. The bifunctionalized isoprenoid was characterized using infrared spectroscopy and (1)H and (13)C nuclear magnetic resonance spectroscopy and was covalently incorporated (0.12 mmol x g(-1)) into the biodegradable aliphatic polyurethane formed on the polycondensation reaction of poly(epsilon-caprolactone) diol, 1,4,3,6-dianhydro-D-sorbitol and 1,6 hexamethylene diisocyanate. Although the covalent incorporation of the isoprenoid molecule into the polyurethane chain modified the surface chemistry of the polymer, it did not affect the viability of attached chondrocytes. Porous 3D scaffolds were produced from the modified and unmodified biodegradable segmented polyurethanes by a salt leaching-phase-inverse technique. The scaffolds were seeded with bovine chondrocytes encapsulated in fibrin gel and cultured in vitro for 14 days. The incorporation of bifunctional isoprenoid into the polyurethane affected the morphology of the scaffolds produced, when compared with the morphology of the scaffolds produced using the same technique from the unmodified polyurethane. As a consequence, there was more uniform cell seeding and more homogeneous distribution of the synthesized extracellular matrix throughout the scaffold resulting in a reduced cell/tissue layer at the edges of the constructs. However, glycosaminoglycan (GAG), DNA content, and chondrocytes phenotype in the scaffolds produced from these two polyurethane formulations did not vary significantly. The findings suggest that the change of surface characteristics and the more open pore structure of the scaffolds produced from the isoprenoid modified polyurethane are beneficial for the seeding efficiency and the homogeneity of the tissue engineered constructs. PMID- 19191319 TI - Biocompatibility and safety evaluation of a ricinoleic acid-based poly(ester anhydride) copolymer after implantation in rats. AB - The aim of this study was to evaluate the safety and tissue compatibility of an injectable biodegradable poly(ester-anhydride) copolymer of ricinoleic acid (RA) and sebacic acid (SA) in rats. The absorbable biomaterial containing 70% w/w of RA and 30% w/w of SA [P(SA-RA) 3:7] was implanted in rats in two separate studies: (1) at high doses subcutaneously (SC) and intramuscularly (IM) simultaneously into the same animal and (2) intracranially (IC). The safety was established in the high-dose administration experiment. No systemic tissue damage, polymer-related lesions, or abnormalities could be detected in the animals. The histopathological evaluation of the SC and IM P(SA-RA) 3:7 implanted sites suggested a typical foreign body reaction (FBR) to biomaterials, and was characterized by excellent tissue repair and good tissue tolerance. In the second experiment, no neurological deficits or behavior changes suggestive of systemic or localized toxicity were observed in the animals implanted IC with the polymer. Only minimal, well-demarcated inflammatory response was observed on days 14 and 21 and consisted of glia cells. No abnormalities were noted in the brain tissue parenchyma located further from the edges of the implant. These results demonstrated that the P(SA-RA) 3:7 copolymer was tolerated well by the animals and compatible with rat subcutaneous, muscle and brain tissues. The biodegradable polymeric system described here could be used as a scaffold for varied applications in localized and sustained delivery of therapeutic agents. PMID- 19191320 TI - Systematic evaluation of the effect of common SNPs on pre-mRNA splicing. AB - The evolutionary and biomedical importance of differential mRNA splicing is well established. Numerous studies have assessed patterns of differential splicing in different genes and correlated these patterns to the genotypes for adjacent single-nucleotide polymorphisms (SNPs). Here, we have chosen a reverse approach and screened dbSNP for common SNPs at either canonical splice sites or exonic splice enhancers (ESEs) that would be classified as putatively splicing-relevant by bioinformatic tools. The 223 candidate SNPs retrieved from dbSNP were experimentally tested using a previously established panel of 92 matching DNAs and cDNAs. For each SNP, 16 cDNAs providing a balanced representation of the genotypes at the respective SNP were investigated by nested RT-PCR and subsequent sequencing. Putative allele-dependent splicing was verified by the cloning of PCR products. The positive predictive value of the bioinformatics tools turned out to be low, ranging from 0% for ESEfinder to 9% (in the case of acceptor-site SNPs) for a recently reported neural network. The results highlight the need for a better understanding of the sequence characteristics of functional splice-sites to improve our ability to predict in silico the splicing relevance of empirically observed DNA sequence variants. PMID- 19191321 TI - Interaction between PHOX2B and CREBBP mediates synergistic activation: mechanistic implications of PHOX2B mutants. AB - Congenital central hypoventilation syndrome (CCHS) is a disorder of the autonomic nervous system. The genetic defect is usually caused by heterozygous mutations of the PHOX2B gene such as a 20-alanine tract (+5 to+13 alanines) expansion (approximately 95%) and occasional frameshift or missense mutations. Cytoplasmic aggregates were shown in PHOX2B proteins with longer alanine tract (+9 and longer) expansion and impaired DNA binding was observed in PHOX2B proteins with frameshift, missense, or longer alanine tract (+9 and longer) expansion. Defective transactivation activity was shown in certain PHOX2B mutants. However, PHOX2B proteins with short alanine tract (+5 to+7) expansion in the majority of patients (approximately 75%) did not have cytoplasmic aggregates or DNA binding defects. CREB-binding protein (CREBBP/CBP) is a transcriptional coactivator that interacts with multiple transcription factors to cause synergistic activation. Here we show that CBP interacted with PHOX2B and served as its coactivator to mediate synergistic activation. Wild-type PHOX2B and CBP used specific domains to interact with each other. The domains of CBP that interacted with different PHOX2B mutants were different compared to those interacting with wild-type PHOX2B. Transient cotransfection assays using different PHOX2B mutants and CBP showed the impaired synergistic activation caused by different PHOX2B mutants. An interfering effect was observed in certain PHOX2B mutants. These results demonstrated that aberrant interaction of PHOX2B mutants with CBP and/or an interfering effect of certain PHOX2B mutants may be the critical mechanism to impair synergistic activation, thereby contributing to the phenotypes of CCHS. PMID- 19191322 TI - The SAAPdb web resource: a large-scale structural analysis of mutant proteins. AB - The Single Amino Acid Polymorphism database (SAAPdb) is a new resource for the analysis and visualization of the structural effects of mutations. Our analytical approach is to map single nucleotide polymorphisms (SNPs) and pathogenic deviations (PDs) to protein structural data held within the Protein Data Bank. By mapping mutations onto protein structures, we can hypothesize whether the mutant residues will have any local structural effect that may "explain" a deleterious phenotype. Our prior work used a similar approach to analyze mutations within a single protein. An analysis of the contents of SAAPdb indicates that there are clear differences in the sequence and structural characteristics of SNPs and PDs, and that PDs are more often explained by our structural analysis. This mapping and analysis is a useful resource for the mutation community and is publicly available at http://www.bioinf.org.uk/saap/db/. PMID- 19191323 TI - Hereditary pancreatitis caused by mutation-induced misfolding of human cationic trypsinogen: a novel disease mechanism. AB - We investigated the biochemical properties and cellular expression of the c.346C>T (p.R116C) human cationic trypsinogen (PRSS1) mutant, which we identified in a German family with autosomal dominant hereditary pancreatitis. This mutation leads to an unpaired Cys residue with the potential to interfere with protein folding via incorrect disulfide bond formation. Recombinantly expressed p.R116C trypsinogen exhibited a tendency for misfolding in vitro. Biochemical analysis of the correctly folded, purified p.R116C mutant revealed unchanged activation and degradation characteristics compared to wild type trypsinogen. Secretion of mutant p.R116C from transfected 293T cells was reduced to approximately 20% of wild type. A similar secretion defect was observed with another rare PRSS1 variant, p.C139S, whereas mutants p.A16V, p.N29I, p.N29T, p.E79K, p.R122C, and p.R122H were secreted normally. All mutants were detected in cell extracts at comparable levels but a large portion of mutant p.R116C was present in an insoluble, protease-sensitive form. Consistent with intracellular retention of misfolded trypsinogen, the endoplasmic reticulum (ER) stress markers immunoglobulin-binding protein (BiP) and the spliced form of the X-box binding protein-1 (XBP1s) were elevated in cells expressing mutant p.R116C. The results indicate that mutation-induced misfolding and intracellular retention of human cationic trypsinogen causes hereditary pancreatitis in carriers of the p.R116C mutation. ER stress triggered by trypsinogen misfolding represents a new potential disease mechanism for chronic pancreatitis. PMID- 19191325 TI - Identification of mutations in the ribosomal protein L5 (RPL5) and ribosomal protein L11 (RPL11) genes in Czech patients with Diamond-Blackfan anemia. AB - Diamond-Blackfan anemia (DBA) is a congenital red blood cell aplasia that is usually diagnosed during early infancy. Apart from defects in red blood cell maturation, the disorder is also associated with various physical anomalies in 40% of patients. Mutations in the ribosomal protein (RP) S19 are found in 25% of patients, while mutations in other proteins of the small ribosomal subunit--RPS17 and RPS24--have been found in a fraction of patients. Recently, mutations in RPL5, RPL11, and RPL35a of the large ribosomal subunit have also been reported in several DBA patients. Here, we present the identification of mutations in the RPL5 and RPL11 genes in patients from the Czech DBA Registry. Mutations in RPL5 were identified in eight patients from 6 out of 28 families (21.4%), and mutations in RPL11 in two patients from 2 out of 28 families (7.1%). Interestingly, all 10 patients with either an RPL5 or RPL11 mutation exhibited one or more physical anomalies; specifically, thumb anomalies (flat thenar) were always present, while no such anomaly was observed in seven patients with an RPS19 mutation. Moreover, 9 out of 10 patients with either an RPL5 or RPL11 mutation were born small for gestational age (SGA) compared to 3 out of 7 patients from the RPS19-mutated group. These observations may suggest that mutations, at least in RPL5, seem to generally have more profound impact on fetal development than mutations in RPS19. Since RPL5 and RPL11, together with RPL23, are also involved in the MDM2-mediated p53 pathway regulation, we also screened the RPL23 gene for mutations; however, no mutations were identified. PMID- 19191324 TI - An in vitro model for de- and remyelination using lysophosphatidyl choline in rodent whole brain spheroid cultures. AB - The process of demyelination occurring in diseases as multiple sclerosis is usually investigated in animal models. A major drawback of animal models is that only one condition can be tested per animal, necessitating many animals and systemic effects are factors to be considered. The aim of the study was to develop a reproducible in vitro model for de- and remyelination using whole brain spheroid cultures and lysophosphatidyl choline (LPC). In spheroid cultures, single cell suspensions of embryonic day 15 rodent brain cells reaggregate under constant rotation. Three-dimensional contacts form between the central nervous system cell types present. Multilayered myelin is maximal in four-week old cultures. A week of repeated exposure to LPC led to 30% loss of MBP protein concentration and 2',3'-cyclic nucleotide 3'-phosphodiesterase activity measurements in both rat and mouse spheroids and 56% loss in the number of myelin sheets, with partial remyelination after a week of recovery. The number of dividing cells was increased after LPC exposure and oligodendrocytes were shown to be among the dividing cells. Microglia and astrocytes were not affected and neurons were relatively spared. This suggests that LPC toxicity is specific for myelin and oligodendrocytes. LPC toxicity could be decreased using cholesterol and simvastatin, suggesting that LPC works through altering membrane composition. Thus, in different rodent species and using different read-outs, we could reproducibly show de- and remyelination in spheroid cultures after LPC exposure. This model for demyelination with potential for remyelination offers possibilities for testing novel therapies and studying mechanisms of remyelination. PMID- 19191326 TI - Imaging glioma cell invasion in vivo reveals mechanisms of dissemination and peritumoral angiogenesis. AB - Infiltration of cancer cells into normal tissue is a hallmark of malignant gliomas and compromises treatment options. A lack of appropriate models limits the study of this invasion in vivo, which makes it difficult to fully understand its anatomy and the role of dynamic interactions with structures of the normal brain. We developed a novel methodology by utilizing multiphoton laser scanning microscopy (MPLSM) to image the movement of glioma cells deep within the normal brain of live mice in real time. This allowed us to track the invasion of individual RFP-expressing GL261 cells in relation to perfused vasculature or GFP labeled endothelial cells repetitively over days, up to a depth of 0.5 mm. Glioma cells moved faster and more efficiently when the abluminal site of a blood vessel was utilized for invasion. Cells that invaded perivascularly were frequently found next to (a) multiple capillary structures where microvessels run parallel to each other, (b) capillary loops or glomeruloid-like bodies, and (c) dilated capillaries. Dynamic MPLSM for more than 48 h revealed that single invasive glioma cells induced intussusceptive microvascular growth and capillary loop formation, specifically at the microvascular site with which they had contact. As the main tumor grew by cooption of existing brain vessels, these peritumoral vascular changes may create a beneficial environment for glioma growth. In conclusion, our study revealed new mechanisms of peritumoral angiogenesis and invasion in gliomas, providing an explanation for their interdependence. PMID- 19191327 TI - Examining requirement for formation of functional Presenilin proteins and their processing events in vivo. AB - Presenilin (Psn) is a multipass transmembrane protein that functions as the catalytic subunit of gamma-secretase for mediating intramembrane cleavage of type 1 transmembrane proteins. Normally active Psn is in the form of a heterodimer composed by its N-terminal and C-terminal fragments that are generated from a Presenilinase-mediated endoproteolytic cleavage within its large cytosolic loop during assembly of the protease complex. Using the Psn forms that either bypass or disable Presenilinase-mediated endoproteolysis, and a Psn form that has most of the large cytosolic loop deleted, we have established an in vivo system to enable investigations of Psn functional domains in Drosophila. We show that the Presenilinase-mediated endoproteolytic event is not essential for producing Psn activity during animal development, and is regulated by integrity of the large cytosolic loop of Psn in Drosophila. The Psn transgenic flies described here could be applied to a broad range of studies on Psn functioning and its related gamma-secretase activity at any developmental stage. PMID- 19191328 TI - 3D finite element model of biofilm detachment using real biofilm structures from CLSM data. AB - In this work, a three-dimensional model of fluid-structure interactions (FSI) in biofilm systems is developed in order to simulate biofilm detachment as a result of mechanical processes. Therein, fluid flow past the biofilm surface results in a mechanical load on the structure which in turn causes internal stresses in the biofilm matrix. When the strength of the matrix is exceeded parts of the structure are detached. The model is used to investigate the influence of several parameters related to the mechanical strength of the biofilm matrix, Young's modulus, Reynolds number, and biofilm structure on biofilm detachment. Variations in biofilm strength and flow conditions significantly influence the simulation outcome. With respect to structural properties the model is widely independent from a change of Young's modulus. A further result of this work indicates that the change of biofilm structure due to growth or other processes will significantly change the stress distribution in the biofilm and thereby the detachment rate. An increase of the mechanical load by increasing fluid flow results in a flat surface of the remaining biofilm structure. It is concluded that the change of structure during biofilm development is the key determinant in terms of the detachment behavior. PMID- 19191329 TI - Increasing the number of diagnostic mutations in malignant hyperthermia. AB - Malignant hyperthermia (MH) is an autosomal dominant disorder characterized by abnormal calcium homeostasis in skeletal muscle in response to triggering agents. Today, genetic investigations on ryanodine receptor type 1 (RYR1) gene and alpha1 subunit of the dihydropyridine receptor (DHPR) (CACNA1S) gene have improved the procedures associated with MH diagnosis. In approximately 50% of MH cases a causative RYR1 mutation was found. Molecular genetic testing based on RYR1 mutations for MH diagnosis is challenging, because the causative mutations, most of which are private, are distributed throughout the RYR1 gene. A more comprehensive genetic testing procedure is needed. Therefore, we aim to expand the genetic information related to MH and to evaluate the effect of mutations on the MH phenotype. Performing an in-depth mutation screening of the RYR1 transcript sequence in 36 unrelated MH susceptible (MHS) patients, we identified 17 novel, five rare, and eight non-disease-causing variants in 23 patients. The 13 remaining MHS patients presented no known variants, neither in RYR1 nor in the CACNA1S binding regions to RYR1. The 17 novel variants were found to affect highly conserved amino acids and were absent in 100 controls. Excellent genotype phenotype correlations were found by investigating 21 MHS families-a total of 186 individuals. Epstein-Barr virus (EBV) lymphoblastoid cells carrying four of these novel mutations showed abnormal calcium homeostasis. The results of this study contribute to the establishment of a robust genetic testing procedure for MH diagnosis. PMID- 19191330 TI - Aph-1 is required to regulate Presenilin-mediated gamma-secretase activity and cell survival in Drosophila wing development. AB - Aph-1 is a multipass transmembrane protein and an essential component of the Presenilin (Psn)-mediated gamma-secretase complex. During protease assembly, Aph 1 stabilizes the newly synthesized Psn holoprotein to facilitate generation of the active form of Psn, which is a Psn-NTF/Psn-CTF heterodimer produced through a Presenilinase-initiated endoproteolytic cleavage of the Psn holoprotein. Although it is clear that loss of Aph-1 activity leads to failure of Psn heterodimer formation, little is understood about whether Aph-1 plays a role in regulating gamma-secretase activity in addition to assisting Psn maturation. Using various modified Psn forms that do not require endoproteolysis or have a large deletion of the cytosolic loop, we show that in Drosophila Aph-1 is still required for gamma-secretase activity independent of its role in promoting Psn endoproteolysis. In addition, our results indicate that Aph-1 is required to promote cell survival in the wing imaginal disc; aph-1 mutant cells are lost either through cell death or because of a defect in cell proliferation. This function of Aph-1 is independent of its role in regulating gamma-secretase activity, but possibly involves downregulating the activity of uncleaved Psn holoprotein. PMID- 19191331 TI - No association between CALHM1 variation and risk of Alzheimer disease. AB - A polymorphism in the calcium homeostasis modulator 1 gene (CALHM1) has recently been associated with risk of late-onset Alzheimer disease. We examined this variant (rs2986017) in 945 Caucasian Americans with late-onset Alzheimer disease and 875 age-matched Caucasian American controls. No association with risk of late onset Alzheimer disease (p=0.368 for genotypes; p=0.796 for alleles) was observed in our sample. However, a potential modest association of minor allele homozygosity (TT) with an earlier age-at-onset was seen (p=0.034). PMID- 19191332 TI - No association between CALHM1 and risk for Alzheimer dementia in a Belgian population. AB - A non-synonymous polymorphism, rs2986017 (p.P86L), in the newly characterized calcium homeostasis modulator 1 (CALHM1) gene located in the Alzheimer dementia (AD) linkage region on 10q24.33, was reported to increase risk of AD, and affect calcium homeostasis and amyloid beta accumulation. We aimed to investigate the association between this functional polymorphism and AD in an independent study population. We genotyped rs2986017 in 362 Belgian AD patients and 519 ethnically matched control individuals. We found no evidence of association between rs2986017 and risk of disease, nor did we find an effect on onset age. Despite its functional properties, our study suggests the polymorphism does not contribute significantly to AD risk in the Belgian population. PMID- 19191333 TI - Functional characterization of ryanodine receptor (RYR1) sequence variants using a metabolic assay in immortalized B-lymphocytes. AB - Mutations in the RYR1 gene are linked to malignant hyperthermia (MH), central core disease and multi-minicore disease. We screened by DHPLC the RYR1 gene in 24 subjects for mutations, and characterized functional alterations caused by some RYR1 variants. Three novel sequence variants and twenty novel polymorphisms were identified. Immortalized lymphoblastoid cell lines from patients with RYR1 variants and from controls were stimulated with 4-chloro-m-cresol (4-CmC) and the rate of extracellular acidification was recorded. We demonstrate that the increased acidification rate of lymphoblastoid cells in response to 4-CmC is mainly due to RYR1 activation. Cells expressing RYR1 variants in the N-terminal and in the central region of the protein (p.Arg530His, p.Arg2163Pro, p.Asn2342Ser, p.Glu2371Gly and p.Arg2454His) displayed higher activity compared with controls; this could account for the MH-susceptible phenotype. Cell lines harboring RYR1(Cys4664Arg) were significantly less activated by 4-CmC. This result indicates that the p.Cys4664Arg variant causes a leaky channel and depletion of intracellular stores. The functional changes detected corroborate the variants analyzed as disease-causing alterations and the acidification rate measurements as a means to monitor Ca(2+)-induced metabolic changes in cells harboring mutant RYR1 channels. PMID- 19191334 TI - Microarray analyses reveal regional astrocyte heterogeneity with implications for neurofibromatosis type 1 (NF1)-regulated glial proliferation. AB - Numerous studies have suggested that astrocytes in the central nervous system (CNS) exhibit molecular and functional heterogeneity. In this regard, astroglia from different CNS locations express distinct immune system, and neurotransmitter proteins, have varying levels of gap junction coupling and respond differently to injury. However, the relevance of these differences to human disease is unclear. As brain tumors in children arise in specific CNS locations, we hypothesized that regional astroglial cell heterogeneity might partly underlie the propensity for gliomas to arise in these areas. In this study, we performed high-density RNA microarray profiling on astrocytes from postnatal day 1 optic nerve, cerebellum, brainstem, and neocortex. We showed that astroglia from each region are molecularly distinct, and we were able to develop gene expression patterns that distinguish astroglia, but not neural stem cells, from these different brain regions. We next used these microarray data to determine whether brain tumor suppressor genes were differentially expressed in these distinct populations of astroglia. Interestingly, neurofibromatosis type 1 (NF1) gene expression was decreased at both the RNA and protein levels in neocortical astroglia relative to astroglia from the other brain regions. To determine the functional significance of this finding, we found increased astroglial cell proliferation in optic nerve, brainstem, and cerebellum, but not neocortex, following Nf1 inactivation in vitro and in vivo. These findings provide molecular evidence for CNS astroglial cell heterogeneity, and suggest that differences in tumor suppressor gene expression might contribute to the regional localization of human brain tumors. PMID- 19191335 TI - Theiler's virus infection induces TLR3-dependent upregulation of TLR2 critical for proinflammatory cytokine production. AB - Theiler's murine encephalomyelitis virus (TMEV) infection directly induces many proinflammatory genes, including type I interferon (IFN) and a variety of cytokine genes. These virus-induced cytokines are a critical factor in developing TMEV-induced demyelinating disease. We have previously reported that the major activation signal for the cytokine genes is mediated via TLR3. In this study, we describe that TLR2 is upregulated via TLR3 signal and cooperatively participates in the expression of IL-6, IL-1beta, CCL2, and CCL5 genes following TMEV infection. The expression of these genes was significantly impaired in both TLR2 deficient and TLR3-deficient primary astrocytes. However, the induction of type I IFNs was not affected by TLR2 deficiency in the primary cells. TMEV infection led to TLR2-mediated NF-kappaB activation, but not IRF3 or IRF7 activation, critical for type I IFN production. More importantly, TLR3 was required for TMEV-induced early TLR2 upregulation in primary astrocytes leading to the production of TLR2 dependent cytokines such as IL-6. Interestingly, soluble factor(s) produced via TLR2/3-dependent signals appears to be partially associated with the downstream cytokine production. These results indicate that TMEV utilizes TLR3-induced TLR2 to induce inflammatory cytokines, which are critical to the development of immune mediated demyelinating disease. PMID- 19191336 TI - Human bone marrow-derived mesenchymal stem cells induce Th2-polarized immune response and promote endogenous repair in animal models of multiple sclerosis. AB - Cell-based therapies are attractive approaches to promote myelin repair. Recent studies demonstrated a reduction in disease burden in mice with experimental allergic encephalomyelitis (EAE) treated with mouse mesenchymal stem cells (MSCs). Here, we demonstrated human bone marrow-derived MSCs (BM-hMSCs) promote functional recovery in both chronic and relapsing-remitting models of mouse EAE, traced their migration into the injured CNS and assayed their ability to modulate disease progression and the host immune response. Injected BM-hMSCs accumulated in the CNS, reduced the extent of damage and increased oligodendrocyte lineage cells in lesion areas. The increase in oligodendrocytes in lesions may reflect BM hMSC-induced changes in neural fate determination, since neurospheres from treated animals gave rise to more oligodendrocytes and less astrocytes than nontreated neurospheres. Host immune responses were also influenced by BM-hMSCs. Inflammatory T-cells including interferon gamma producing Th1 cells and IL-17 producing Th17 inflammatory cells and their associated cytokines were reduced along with concomitant increases in IL-4 producing Th2 cells and anti inflammatory cytokines. Together, these data suggest that the BM-hMSCs represent a viable option for therapeutic approaches. PMID- 19191337 TI - Accelerating molecular dynamic simulation on graphics processing units. AB - We describe a complete implementation of all-atom protein molecular dynamics running entirely on a graphics processing unit (GPU), including all standard force field terms, integration, constraints, and implicit solvent. We discuss the design of our algorithms and important optimizations needed to fully take advantage of a GPU. We evaluate its performance, and show that it can be more than 700 times faster than a conventional implementation running on a single CPU core. PMID- 19191338 TI - Characterization and expression analysis of a gene cluster for nitrate assimilation from the yeast Arxula adeninivorans. AB - In Arxula adeninivorans nitrate assimilation is mediated by the combined actions of a nitrate transporter, a nitrate reductase and a nitrite reductase. Single copy genes for these activities (AYNT1, AYNR1, AYNI1, respectively) form a 9103 bp gene cluster localized on chromosome 2. The 3210 bp AYNI1 ORF codes for a protein of 1070 amino acids, which exhibits a high degree of identity to nitrite reductases from the yeasts Pichia anomala (58%), Hansenula polymorpha (58%) and Dekkera bruxellensis (54%). The second ORF (AYNR1, 2535 bp) encodes a nitrate reductase of 845 residues that shows significant (51%) identity to nitrate reductases of P. anomala and H. polymorpha. The third ORF in the cluster (AYNT1, 1518 bp) specifies a nitrate transporter with 506 amino acids, which is 46% identical to that of H. polymorpha. The three genes are independently expressed upon induction with NaNO(3). We quantitatively analysed the promoter activities by qRT-PCR and after fusing individual promoter fragments to the phytase (phyK) gene from Klebsiella sp. ASR1. The AYNI1 promoter was found to exhibit the highest activity, followed by the AYNT1 and AYNR1 elements. Direct measurements of nitrate and nitrite reductase activities performed after induction with NaNO(3) are compatible with these results. Both enzymes show optimal activity at around 42 degrees C and near-neutral pH, and require FAD as a co-factor and NADPH as electron donor. PMID- 19191339 TI - Two novel CLCN2 mutations accelerating chloride channel deactivation are associated with idiopathic generalized epilepsy. AB - Heterozygous mutations in the CLCN2 gene encoding the voltage-gated chloride channel CLC2 have been identified in patients with idiopathic generalized epilepsy (IGE). Yet the involvement of CLCN2 in epilepsy remains controversial. To investigate the involvement of CLCN2 in another independent sample, we screened 52 unrelated patients from IGE families and 23 patients with Doose syndrome for mutations in CLCN2. No mutations were found in patients with Doose syndrome. In three unrelated IGE families, we identified two novel missense mutations, p.Arg235Gln and p.Arg577Gln, which were absent in large ethnically matched control populations, and one novel p.Arg644Cys variant, which was also found in five Indian controls. Functional characterization of mutant channels using heterologous expression in mammalian cells and whole-cell patch-clamp recordings revealed faster deactivation kinetics as the major phenotype of both missense mutations. This finding predicts a loss of function that may contribute to intracellular chloride accumulation or neuronal hyperexcitability. However, the incomplete segregation of the mutations among affected members and the transmission by unaffected parents suggests that these CLCN2 mutations alone are not sufficient to induce epilepsy. They may instead represent susceptibility factors among other so far undetected genetic alterations in the respective families. PMID- 19191340 TI - Adherence to antiparkinson medication in a multicenter European study. AB - Two small studies reported suboptimal therapy adherence in Parkinson's disease. We conducted a larger multicenter European study to assess medicine-taking behavior. Parkinson's disease patients taking dopaminergic therapy were enrolled in 8 centers in 5 countries, and disease severity and demographics recorded. Antiparkinson drug adherence was measured for 4 weeks using electronic monitoring bottles which record the date and time of cap opening (Aardex, Switzerland). One hundred twelve patients, mean age 65 years (standard deviation (SD) 10), with Parkinson's disease for 7.7 (SD 8.2) years completed the study. Total median adherence (doses taken/doses prescribed) was 97.7% (interquartile range [IQ] 90.6 100), days adherence (correct dose days) was 86.2% (IQ 61.1-96.2) and timing adherence (doses taken at correct time intervals) was 24.4% (IQ 5.3-56.5). Fourteen patients (12.5%) took less than 80% of prescribed doses, which was defined as suboptimal adherence. Patients with satisfactory adherence took a median of 8 mg/day (IQ 0-33) less than their prescribed dose of levodopa (P = NS), while suboptimal adherence patients took a median of 481 mg/day (IQ 205-670) less than their prescribed dose (P = 0.0006). The Parkinson motor score was significantly higher in patients with suboptimal adherence at 29 (IQ 20-40), versus those with satisfactory adherence at 19 (IQ 13-26), P = 0.005. Once daily drugs had significantly better adherence when compared with drugs prescribed more frequently (P < 0.0001). Suboptimal therapy adherence is associated with significant deviation from prescribed levodopa doses, despite greater Parkinson's motor severity. Optimizing oral medication intake has a potential role in maximizing the therapy response in Parkinson's disease. PMID- 19191341 TI - Various movement disorders in a patient with Sjogren syndrome. PMID- 19191342 TI - Cognitive decline in early Parkinson's disease. AB - Data on the prevalence and severity of cognitive impairment among patients with newly diagnosed idiopathic Parkinson's disease (PD) is limited. Using a prospectively collected clinical database, we studied the longitudinal trend of mini-mental state examination (MMSE) change and baseline factors predictive for MMSE decline. One hundred six patients with mean age of 61.2 years and mean baseline MMSE of 27.8 +/- 2.3 were studied. MMSE increased by 0.4 points/year among patients without cognitive decline (n = 73) and decreased by 2.39 points/year among patients with cognitive decline (n = 33). Univariate analysis demonstrated education, age of diagnosis, depression, and diabetes mellitus to be associated with cognitive decline. Motor scores and hallucination were not associated with cognitive decline. Multivariate analysis demonstrated higher level of education to be protective (HR = 0.91, 95% CI 0.82-0.99, P = 0.047) and depression having borderline significance in predicting cognitive decline (HR = 2.00, 95% CI 0.97-4.15, P = 0.061). We found that 31% of newly diagnosed idiopathic PD patients have measurable cognitive decline at an early stage of disease. Higher education is protective while depression may be predictive of cognitive decline. PMID- 19191343 TI - Levodopa use and sleep in patients with dementia with Lewy bodies. AB - Sleep disturbance and excessive daytime somnolence (EDS) are features of Parkinson's disease (PD) and dementia with Lewy bodies (DLB) that may be influenced by dopamine replacement therapy. The effect of levodopa on sleep and EDS in DLB is unknown and unclear in PD. The aim of this study is to determine if levodopa treatment alters sleep symptoms and EDS in DLB. Dopamine naive patients with DLB (n = 15; mean mini mental state examination (MMSE) score 17.7(4.6)) and PD (n = 9; mean MMSE 25.5(2.2)) were assessed using the Epworth sleep scale, Parkinson's disease sleep scale, and the neuropsychiatric inventory prior to initiating treatment with levodopa. All measures were repeated after 3 and 6 months of levodopa therapy. The median final daily levodopa dose was 300 mg in both groups. Baseline sleep measures were comparable between groups. Levodopa treatment did not affect sleep or lead to increased EDS in DLB patients. The use of levodopa does not appear to adversely affect subjective sleep measures or increase EDS in DLB patients. PMID- 19191344 TI - Facial reflex hyperexcitability in geniospasm suggests a brainstem origin. PMID- 19191345 TI - Predicting behavioral problems in craniopharyngioma survivors after conformal radiation therapy. AB - BACKGROUND: Although radiation therapy is a primary treatment for craniopharyngioma, it can exacerbate existing problems related to the tumor and pre-irradiation management. Survival is often marked by neurologic deficits, panhypopituitarism, diabetes insipidus, cognitive deficiencies, and behavioral and social problems. PROCEDURE: The Achenbach Child Behavior Checklist (CBCL) was used to evaluate behavioral and social problems during the first 5 years of follow-up in 27 patients with craniopharyngioma treated with conformal radiation therapy. RESULTS: All group averages for the CBCL scales were within the age typical range at pre-irradiation baseline. Extent of surgical resection was implicated in baseline differences for the internalizing, externalizing, behavior problem and social scores. Significant longitudinal changes were found in internalizing, externalizing, behavior problem and school scores that correlated with tumor and treatment-related factors. CONCLUSIONS: The most common variables implicated in post-irradiation behavioral and social problems were CSF shunting, presence of an Ommaya reservoir, diabetes insipidus, and low pre-irradiation growth hormone levels. PMID- 19191346 TI - Ex vivo-expanded donor CD4(+) lymphocyte infusion against relapsing neuroblastoma: A transient graft-versus-tumor effect. AB - High-risk neuroblastoma has a poor prognosis despite multimodal treatment including high-dose chemotherapy. A 7-year-old male with neuroblastoma received ex vivo-expanded donor CD4(+) T lymphocyte infusion (CD4(+) DLI) after recurrence in the bone marrow following allogeneic hematopoietic stem cell transplantation from his HLA-identical mother. The disease transiently responded to CD4(+) DLI with reduction of tumor cells and a decrease of serum neuron-specific enolase. The response was associated with development of continued high fever and an increase of cytotoxic T lymphocytes in peripheral blood. This case suggests a possibility of a graft-versus-tumor effect against neuroblastoma. PMID- 19191348 TI - Cognitive function and common mental disorders in older people with vascular and non-vascular disorders: a national survey. AB - OBJECTIVE: Vascular disorders are important potential causes of cognitive impairment and common mental disorders but their specificity as risk factors has yet to be clarified. The objectives of this analysis were to compare vascular and non-vascular health problems with respect to their associations with cognitive function and common mental disorder. METHOD: An analysis was carried out of cross sectional data from the 2000 UK National Survey of Psychiatric Morbidity. The two dependent variables analysed were common mental disorder (revised Clinical Interview Schedule) and cognitive function (TICSm and animal naming) in survey participants who were aged 60-74 years (n = 2007). Associations with self reported vascular and common non-vascular (musculoskeletal, respiratory or gastrointestinal) disorders were compared. Disability (SF-12 physical scale) was considered as a potential confounding/mediating factor). RESULTS: Vascular disorders were associated with impaired global cognitive function and lower memory but not verbal fluency scores after adjustment for age, gender, education and social class. No such associations were found for non-vascular disorders. Vascular and non-vascular disorders were associated with common mental disorder to a similar extent, and associations were substantially explained by disability. CONCLUSIONS: Lower cognitive function was specifically associated with vascular disorders. Findings for common mental disorder were more consistent with an effect of disability rather than the actual consequences of specific health complaints. PMID- 19191349 TI - Paroxysmal dystonia associated to primary Sjogren's syndrome. PMID- 19191347 TI - Dual regulation of Ca2+-dependent glutamate release from astrocytes: vesicular glutamate transporters and cytosolic glutamate levels. AB - Vesicular glutamate transporters (VGLUTs) are responsible for vesicular glutamate storage and exocytotic glutamate release in neurons and astrocytes. Here, we selectively and efficiently overexpressed individual VGLUT proteins (VGLUT1, 2, or 3) in solitary astrocytes and studied their effects on mechanical stimulation induced Ca2+-dependent glutamate release. Neither VGLUT1 nor VGLUT2 overexpression changed the amount of glutamate release, whereas overexpression of VGLUT3 significantly enhanced Ca2+-dependent glutamate release from astrocytes. None of the VGLUT overexpression affected mechanically induced intracellular Ca2+ increase. Inhibition of glutamine synthetase activity by L-methionine sulfoximine in astrocytes, which leads to increased cytosolic glutamate concentration, greatly increased their mechanically induced Ca2+-dependent glutamate release, without affecting intracellular Ca2+ dynamics. Taken together, these data indicate that both VGLUT3 and the cytosolic concentration of glutamate are key limiting factors in regulating the Ca2+-dependent release of glutamate from astrocytes. PMID- 19191350 TI - Secondary follicle growth and oocyte maturation by culture in alginate hydrogel following cryopreservation of the ovary or individual follicles. AB - An option for fertility preservation for women facing a cancer diagnosis involves the cryopreservation of ovarian tissue for later re-transplantation or in vitro culture, with in vitro culture preferred to avoid reintroduction of the cancer. Small, immature follicles survive the freeze-thaw process, and can be matured through in follicle maturation (IFM) that involves an initial growth of the follicle and subsequent maturation of the oocyte. The ovarian tissue can be cryopreserved in two forms: (i) cortical strips consisting of follicles and surrounding stroma (Cryo-Ov) or (ii) individually isolated follicles (Cryo-In). The aim of this study was to assess the follicle growth and oocyte maturation for follicles that were cryopreserved either as strips or individually using a slow freezing cryopreservation method. The two follicle groups, together with non cryopreserved control follicles, were grown in an alginate-based three dimensional culture system for 12 days. The overall survival, size increase and antrum formation rates were comparable among the three groups. At day 12 of culture, Androstenedione levels were decreased in the Cryo-Ov group relative to the other two, and the ratio of progesterone to estradiol was increased in the two cryopreserved groups relative to the control. Both Gja1 (known as connexin 43) and Gja4 (known as connexin 37) mRNA expression were decreased at day 6 in the cryopreserved groups relative to controls, and by day 12, Gja1 was similar for all three groups. Moreover, Cryo-In resulted in lower GVBD rate indicating some impaired oocyte development. Overall, the present study demonstrated that mouse preantral follicles, either within ovarian tissues or individually isolated, could be successfully cryopreserved by the slow-freezing method, as evidenced by post-thaw follicle development and steroidgenesis, oocyte maturation and molecular markers for oocyte and/or granulosa cells connection. PMID- 19191351 TI - Effects of energy dissipation rate on islets of Langerhans: implications for isolation and transplantation. AB - Acute physical stresses can occur in the procurement and isolation process and potentially can contribute to islet death or malfunction upon transplantation. A contractional flow device, previously used to subject suspended cells to well defined hydrodynamic forces, has been modified and used to assess the vulnerability of porcine islets of Langerhans to hydrodynamic forces. The flow profiles and velocity gradients in this modified device were modeled using commercial CFD software and characterized, as in previous studies, with the scalar parameter, energy dissipation rate (EDR). Porcine islets were stressed in a single pass at various stress levels (i.e., values of EDR). Membrane integrity, oxygen uptake rate, caspase 3/7 activity, and insulin release were not affected by the levels of fluid stress tested up to an EDR of 2 x 10(3) W/m(3). Visual observation of the stressed islets suggested that cells at the islet exterior were peeled away at EDR greater than 10,000 W/m(3), however, this observation could not be confirmed using image analysis software, which determined the ratio of surface perimeter to total area. The result of this study suggests an upper limit in fluid stress to which islets can be subjected. Such upper limits assist in the design and operation of future islet processing equipment and processes. PMID- 19191352 TI - Secondary flow mixing due to biofilm growth in capillaries of varying dimensions. AB - Using a magnetic resonance microscopy (MRM) technique, velocity perturbations due to biofouling in capillaries were detected in 3D velocity maps. The velocity images in each of the three square capillary sizes (2, 0.9, and 0.5 mm i.d.) tested indicate secondary flow in both the x- and y-directions for the biofouled capillaries. Similar flow maps generated in a clean square capillary show only an axial component. Investigation of these secondary flows and their geometric and dynamic similarity is the focus of this study. The results showed significant secondary flows present in the 0.9 mm i.d. capillary, on the scale of 20% of the bulk fluid flow. Since this is the "standard 1 mm" size capillary used in confocal microscopy laboratory bioreactors to investigate biofilm properties, it is important to understand how these enhanced flows impact bioreactor transport. PMID- 19191353 TI - Syntrophic interactions among anode respiring bacteria (ARB) and Non-ARB in a biofilm anode: electron balances. AB - We demonstrate that the coulombic efficiency (CE) of a microbial electrolytic cell (MEC) fueled with a fermentable substrate, ethanol, depended on the interactions among anode respiring bacteria (ARB) and other groups of micro organisms, particularly fermenters and methanogens. When we allowed methanogenesis, we obtained a CE of 60%, and 26% of the electrons were lost as methane. The only methanogenic genus detected by quantitative real-time PCR was the hydrogenotrophic genus, Methanobacteriales, which presumably consumed all the hydrogen produced during ethanol fermentation ( approximately 30% of total electrons). We did not detect acetoclastic methanogenic genera, indicating that acetate-oxidizing ARB out-competed acetoclastic methanogens. Current production and methane formation increased in parallel, suggesting a syntrophic interaction between methanogens and acetate-consuming ARB. When we inhibited methanogenesis with 50 mM 2-bromoethane sulfonic acid (BES), the CE increased to 84%, and methane was not produced. With no methanogenesis, the electrons from hydrogen were converted to electrical current, either directly by the ARB or channeled to acetate through homo-acetogenesis. This illustrates the key role of competition among the various H(2) scavengers and that, when the hydrogen-consuming methanogens were present, they out-competed the other groups. These findings also demonstrate the importance of a three-way syntrophic relationship among fermenters, acetate-consuming ARB, and a H(2) consumer during the utilization of a fermentable substrate. To obtain high coulombic efficiencies with fermentable substrates in a mixed population, methanogens must be suppressed to promote new interactions at the anode that ultimately channel the electrons from hydrogen to current. PMID- 19191355 TI - SERS labels for red laser excitation: silica-encapsulated SAMs on tunable gold/silver nanoshells. AB - In a glass house: Silica-encapsulated self-assembled monolayers (SAMs) on tunable gold/silver nanoshells were used as labels for surface-enhanced Raman scattering (SERS). This concept combines the spectroscopic advantages arising from maximum surface coverage and uniform molecular orientation of the Raman reporter molecules within the complete monolayer together with the high chemical and mechanical stability of the glass shell. PMID- 19191354 TI - Structural genomics reveals EVE as a new ASCH/PUA-related domain. AB - We report on several proteins recently solved by structural genomics consortia, in particular by the Northeast Structural Genomics consortium (NESG). The proteins considered in this study differ substantially in their sequences but they share a similar structural core, characterized by a pseudobarrel five stranded beta sheet. This core corresponds to the PUA domain-like architecture in the SCOP database. By connecting sequence information with structural knowledge, we characterize a new subgroup of these proteins that we propose to be distinctly different from previously described PUA domain-like domains such as PUA proper or ASCH. We refer to these newly defined domains as EVE. Although EVE may have retained the ability of PUA domains to bind RNA, the available experimental and computational data suggests that both the details of its molecular function and its cellular function differ from those of other PUA domain-like domains. This study of EVE and its relatives illustrates how the combination of structure and genomics creates new insights by connecting a cornucopia of structures that map to the same evolutionary potential. Primary sequence information alone would have not been sufficient to reveal these evolutionary links. PMID- 19191356 TI - What does it really take to stabilize complexes of late transition metals with terminal oxo ligands? AB - Surprisingly stable: Noble-metal complexes with terminal oxo ligands are frequently postulated as intermediates, but they are generally considered elusive, as their d electrons destabilize the M=O units. Until recently, the isolation of such compounds was thought to require strong acceptor ligands, but now a remarkably stable Pt=O complex has been obtained employing a simple pincer ligand. PMID- 19191357 TI - Stereogradient polymers formed by controlled/living radical polymerization of bulky methacrylate monomers. AB - Life RAFT: A bulky methacrylate monomer, triphenylmethyl methacrylate (TrMA), was polymerized with reversible addition-fragmentation chain transfer (RAFT) agents. Stereogradient polymers in which the isospecificity increased spontaneously as the monomer concentration decreased were formed by a polymerization depolymerization equilibrium that can convert a less stable growing polymer terminal into a more stable form (see picture). PMID- 19191359 TI - Birch reduction of benzene in a low-temperature plasma. AB - Selective and specific dihydrogenation of benzene and other arenes has been observed in a low-temperature helium plasma. A surface Birch reduction mechanism has been proposed in which benzene molecules adsorbed on the discharge surface capture low-energy surface-adsorbed electrons and subsequently undergo protonation (see picture). Gas-phase oxidation processes accompany the reduction reaction. PMID- 19191358 TI - Pattern-based recognition of heparin contaminants by an array of self-assembling fluorescent receptors. AB - Tracking down potential killers: Strong host-guest interactions enable the facile combination of polycationic cyclodextrin binding motifs (blue) with fluorescent reporters (orange) tethered to a hydrophobic guest molecule (dark green). An array of supramolecular fluorescent receptors prepared by this modular approach was used for the pattern-based recognition of negatively charged contaminants in the anticoagulant drug heparin. PMID- 19191360 TI - A stable and crystalline triarylgermyl radical: structure and EPR spectra. AB - A radical thing: After being obtained unexpectedly in low yields, the synthesis of the first stable triarylgermyl radical (*)Ge[3,5-tBu(2)-2,6-(EtO)(2)C(6)H](3) (1; C gray, O blue) was considerably optimized, and the product was investigated by X-ray analysis and EPR spectroscopy. The results were compared with DFT-MO studies for the model compound (*)Ge[2,6-(MeO)(2)C(6)H(3)]. PMID- 19191361 TI - [Co@Ge10]3-: an intermetalloid cluster with archimedean pentagonal prismatic structure. AB - Inorganic pentaprismane: The unusual structure of the anion [Co@Ge(10)](3-), which was obtained by the reaction of K(4)Ge(9) with [Co(C(8)H(12))(C(8)H(13))] in ethylenediamine, raises questions about chemical bonding in the anion. The Zintl ion cluster has virtual D(5h) symmetry and is a unique example of a ligand free cluster that is not a deltahedron. The delocalized chemical bonding is represented in the picture by one of the bonding orbitals of the anion. PMID- 19191362 TI - Nanoparticle-enhanced fluorescence imaging of latent fingerprints reveals drug abuse. AB - No sweat! The sweat in a latent fingerprint (LFP) can contain orally ingested drugs and their metabolites. In a new method for drug detection, primary antibodies (Abs) against drug metabolites are conjugated to magnetic nanoparticles (NPs). The LFP is incubated with the NPs, excess particles removed, and the LFP treated with a fluorescently labeled secondary antibody. Fluorescence imaging then allows characterization. PMID- 19191363 TI - Zn(1.2)H(0.6)PW(12)O(40) Nanotubes with double acid sites as heterogeneous catalysts for the production of biodiesel from waste cooking oil. AB - Out of the frying pan: A ZnPW nanotube catalyst containing Bronsted and Lewis double acid sites promotes the conversion of waste cooking oil into biodiesel. The catalytic activity of the ZnPW nanotubes is stable to the presence of free fatty acids or water in the feedstock. The high catalytic activity of the ZnPW nanotubes is attributed to the synergistic effect of Lewis acid sites and Bronsted acid sites.Zinc dodecatungstophosphate (Zn(1.2)H(0.6)PW(12)O(40); ZnPW) nanotubes, which feature Lewis acid and Bronsted acid sites, were prepared using cellulose fibers as templates. The structure, acid properties, and catalytic activity of the nanotubes as heterogeneous catalysts for biodiesel production were then studied in detail. The ZnPW nanocatalyst exhibited higher catalytic activities for the simultaneous esterification and transesterification of palmitic acid than the parent acid catalyst 12-tungstophosphoric acid (H(3)PW(12)O(40)). Moreover, the doubly acidic nanotubes led to markedly enhanced yields of methyl esters in the conversion of waste cooking oil (containing 26.89 wt % free fatty acids and 1 % moisture) to biodiesel. The catalyst could be recycled and reused with negligible loss in activity over five cycles. The ZnPW nanocatalyst is acid- and water-tolerant and is an environmentally benign heterogeneous catalyst for the production of biodiesel from low-quality feedstocks. PMID- 19191364 TI - Giant pores in a chromium 2,6-naphthalenedicarboxylate open-framework structure with MIL-101 topology. AB - Large, larger, ... Replacement of 1,4-benzenedicarboxylate by 2,6 naphthalenedicarboxylate in the MIL-101 structure leads to an isoreticular mesoporous framework containing cages with diameters of 39 and 46 A. High throughput methods are employed to determine appropriate reaction conditions. The microcrystalline compound is characterized by molecular simulation techniques, powder X-ray diffraction, N(2)-sorption, and TEM investigations. PMID- 19191365 TI - Ligand-stabilized chalcogen dications. AB - Chalcogen-transfer reagents? The bonding in the dicationic rings C(2)N(2)E(2+) (see picture) differs from that in N-heterocyclic carbenes and their isovalent p block analogues in accommodating a lone pair of electrons with pi symmetry, as well as sigma symmetry, on the chalcogen center. The labile electrophilic chalcogenium dications (E(2+)) are potentially versatile chalcogen-transfer reagents in reactions with a variety of inorganic and organic substrates. PMID- 19191366 TI - Intermolecular methoxycarbonylation of terminal alkynes catalyzed by palladium(II) bis(oxazoline) complexes. AB - Boxing clever: Direct conversion of a terminal alkyne group into a beta methoxyacrylate is realized with the aid of the bis(oxazoline) ligand (box). Acetyl and ketal protecting groups, free hydroxy groups, and acid-sensitive glycosidic bonds are not affected under the reaction conditions. The one-pot synthesis of (+/-)-dihydrokawain from the homopropargyl alcohol is also achieved. tfa = trifluoroacetate. PMID- 19191368 TI - Force-clamp spectroscopy of reversible bond breakage. AB - We consider reversible breaking of adhesion bonds or folding of proteins under the influence of a constant external force. We discuss the statistical properties of the unbinding/rebinding events and analyze their mean number and their variance in the framework of simple kinetic models. In the calculations, we explicitly exploit the analogy to single molecule fluorescence and particularly between unbinding/rebinding and photon emission events. Whereas for two-state behavior Poisson or sub-Poisson statistics of the events is found, we show that for more general kinetic schemes also super-Poisson statistics can occur. Temporal fluctuations of the transition rates, a hallmark for the presence of dynamic disorder, should become experimentally accessible via the determination of the second moment of the event-number distribution. PMID- 19191369 TI - Production of vibrationally excited H(2)O from charge exchange of H(3)O(+) with cesium. AB - The center-of-mass kinetic energy release for the dissociation of H(3)O following charge exchange of H(3)O(+) with cesium has been studied experimentally and modeled using quasiclassical trajectory calculations based on an ab initio potential energy surface for H(3)O(+) and "direct dynamics" for H(3)O. Branching fractions for the H(2)O+H and OH+H(2) dissociation channels have been measured and compared to the calculations. The dominant channel is found to be H(2)O+H and the experimental kinetic energy release spectrum reveals that H(2)O is formed with a vibrational inversion in stretching vibrations, coupled with low bending and rotational excitation. PMID- 19191370 TI - Semiclassical on-the-fly computation of the S(0)-->S(1) absorption spectrum of formaldehyde. AB - The anharmonic S(0)-->S(1) vibronic absorption spectrum of the formaldehyde molecule is computed on the fly using semiclassical dynamics. This first example of an on-the-fly semiclassical computation of a vibronic spectrum was achieved using a unit prefactor modified frozen Gaussian semiclassical propagator for the excited state. A sample of 6000 trajectories sufficed for obtaining a converged spectrum, which is in reasonable agreement with experiment. Similar agreement is not obtained when using a harmonic approximation for the spectrum, demonstrating the need for a full anharmonic computation. This first example provides a resolution of approximately 100 cm(-1). Potential ways of improving the methodology and obtaining higher resolution and accuracy are discussed. PMID- 19191371 TI - Coarse-grained models for fluids and their mixtures: Comparison of Monte Carlo studies of their phase behavior with perturbation theory and experiment. AB - The prediction of the equation of state and the phase behavior of simple fluids (noble gases, carbon dioxide, benzene, methane, and short alkane chains) and their mixtures by Monte Carlo computer simulation and analytic approximations based on thermodynamic perturbation theory is discussed. Molecules are described by coarse grained models, where either the whole molecule (carbon dioxide, benzene, and methane) or a group of a few successive CH(2) groups (in the case of alkanes) are lumped into an effective point particle. Interactions among these point particles are fitted by Lennard-Jones (LJ) potentials such that the vapor liquid critical point of the fluid is reproduced in agreement with experiment; in the case of quadrupolar molecules a quadrupole-quadrupole interaction is included. These models are shown to provide a satisfactory description of the liquid-vapor phase diagram of these pure fluids. Investigations of mixtures, using the Lorentz-Berthelot (LB) combining rule, also produce satisfactory results if compared with experiment, while in some previous attempts (in which polar solvents were modeled without explicitly taking into account quadrupolar interaction), strong violations of the LB rules were required. For this reason, the present investigation is a step towards predictive modeling of polar mixtures at low computational cost. In many cases Monte Carlo simulations of such models (employing the grand-canonical ensemble together with reweighting techniques, successive umbrella sampling, and finite size scaling) yield accurate results in very good agreement with experimental data. Simulation results are quantitatively compared to an analytical approximation for the equation of state of the same model, which is computationally much more efficient, and some systematic discrepancies are discussed. These very simple coarse-grained models of small molecules developed here should be useful, e.g., for simulations of polymer solutions with such molecules as solvent. PMID- 19191372 TI - An arbitrary order Douglas-Kroll method with polynomial cost. AB - A new Douglas-Kroll transformation scheme up to arbitrary order is presented to study the convergence behavior of the Douglas-Kroll series and the influence of different choices of parametrization for the unitary transformation. The standard approach for evaluating the Douglas-Kroll Hamiltonian suffers from computational difficulties due to the huge number of matrix multiplications, which increase exponentially with respect to the order of truncation. This makes it prohibitively expensive to obtain results for very high order Douglas-Kroll Hamiltonians. The highest order previously presented is 14th order, but it is not enough to obtain accurate results for systems containing heavy elements, where the Douglas-Kroll series converges very slowly. In contrast, our approach dramatically reduces the number of matrix multiplications, which only increase with a polynomial scaling. With the new method, orders greater than 100 and machine accuracy are possible. This fast method is achieved by employing a special transformation to all Douglas-Kroll operators and our algorithm is very simple. We demonstrate the performance of our implementation with calculations on one-electron systems and many-electron atoms. All results show very good convergence behavior of the Douglas-Kroll series. Very small differences are found between the different parametrizations, and therefore the exponential form, which is the simplest and fastest, is recommended. PMID- 19191373 TI - Double spin-flip approach within equation-of-motion coupled cluster and configuration interaction formalisms: Theory, implementation, and examples. AB - The spin-flip (SF) approach is extended to excitations that flip the spin of two electrons to describe multiconfigurational M(s)=0 wave functions via high spin quintet references. Equations and implementation of the double SF (2SF) approach within equation-of-motion coupled-cluster (EOM-CC) and configuration interaction (CI) formalisms are presented. The numerical performance of the resulting EOM-2SF CC and 2SF-CI models is demonstrated by calculations of symmetric dissociation of O-H bonds in water, electronic states of linear H(4), double CC bond-breaking in ethylene, and low-lying states of trimethylenemethyl diradical and 2,4 didehydrometaxylylene tetraradical. The results of active-space variants of 2SF are very close to the more computationally expensive full-space counterparts. An efficient implementation of the active-space approximation of the 2SF configuration interaction doubles (CID) model termed 2SF-configuration interaction singles (CIS) is also reported. The scaling of 2SF-CIS is only N(4), which allows applications to relatively large molecules. PMID- 19191375 TI - Excitonic versus electronic couplings in molecular assemblies: The importance of non-nearest neighbor interactions. AB - We demonstrate that for a range of phenylene- and thiophene-based conjugated polymers of practical relevance for optoelectronic applications, exciton couplings in one-dimensional stacks deviate significantly from the nearest neighbor approximation. Instead, long-range interactions with non-nearest neighbors have to be included, which become increasingly important with growing oligomer size. While the exciton coupling vanishes for infinitely long ideal polymer chains and provides a sensitive measure of the actual conjugation length, the electronic coupling mediating charge transport shows rapid convergence with molecular size. Similar results have been obtained for very different molecular backbones, thus highlighting the general character of these findings. PMID- 19191374 TI - Effect of an external electric field on the dissociation energy and the electron density properties: The case of the hydrogen bonded dimer HF...HF. AB - The effect of a homogeneous external electric field parallel to the hydrogen bond in the FH...FH dimer has been studied by theoretical methods. The quantum theory of atoms in molecules methodology has been used for analyzing the electron distribution of the dimer, calculated with different hydrogen bond distances and external field magnitudes. It is shown that an electric field in the opposite direction to the dipole moment of the system strengthens the interaction due to a larger mutual polarization between both molecules and increases the covalent character of the hydrogen bond, while an external field in the opposite direction has the inverse effect. The properties of the complex at its equilibrium geometry with applied field have been calculated, showing that dependencies between hydrogen bond distance, dissociation energy, and properties derived from the topological analysis of the electron distribution are analogous to those observed in families of XDH...AY complexes. The application of an external field appears as a useful tool for studying the effect of the atomic environment on the hydrogen bond interaction. In the case of FH...FH, both the kinetic energy density and the curvature of the electron density along the hydrogen bond at the bond critical point present a surprisingly good linear dependence on the dissociation energy. The interaction energy can be modeled by the sum of two exponential terms that depend on both the hydrogen bond distance and the applied electric field. Moreover, as indicated by the resulting interaction energy observed upon application of different external fields, the equilibrium distance varies linearly with the external field, and the dependence of the dissociation energy on either the hydrogen bond distance or the external electric field is demonstrated to be exponential. PMID- 19191376 TI - Analytic expressions for electrical energy and electrical force of two spheres. AB - Analytical expressions for the electrical force and the electrical energy of two spheres immersed in a symmetric electrolyte solution are derived under conditions of constant surface potential and constant surface charge. Previous analysis under Debye-Huckel condition (linear case) is extended to the corresponding nonlinear case; because the level of the surface potential is arbitrary the results obtained have much wider applications. In general, the performance of the analytical formulas derived is satisfactory, and better than that of the available results in literature. For the case of constant surface potential, the analytic formulas obtained are most accurate for large particles, and for the case of constant surface charge, they are most accurate for small particles. PMID- 19191377 TI - A systematic understanding of orbital energy shift in polar solvent. AB - The orbital energy of molecule is significantly shifted upon going from gas phase to solution phase. Based on Koopmans' theorem, the shift should be related to the change of ionization potential. However, the computed shift looks usually random and clear understanding has not been attained yet. Furthermore it is obtained only after solving complicated equations. In this study, we report a systematic framework for understanding the orbital energy shift by solvation effect and simple approximate formulae are presented. PMID- 19191378 TI - Double excitations in finite systems. AB - Time-dependent density-functional theory (TDDFT) is widely used in the study of linear response properties of finite systems. However, there are difficulties in properly describing excited states, which have double- and higher-excitation characters, which are particularly important in molecules with an open-shell ground state. These states would be described if the exact TDDFT kernel were used; however, within the adiabatic approximation to the exchange-correlation (xc) kernel, the calculated excitation energies have a strict single-excitation character and are fewer than the real ones. A frequency-dependent xc kernel could create extra poles in the response function, which would describe states with a multiple-excitation character. We introduce a frequency-dependent xc kernel, which can reproduce, within TDDFT, double excitations in finite systems. In order to achieve this, we use the Bethe-Salpeter equation with a dynamically screened Coulomb interaction W(omega), which can describe these excitations, and from this we obtain the xc kernel. Using a two-electron model system, we show that the frequency dependence of W does indeed introduce the double excitations that are instead absent in any static approximation of the electron-hole screening. PMID- 19191379 TI - Enthalpy of formation and anharmonic force field of diacetylene. AB - The enthalpy of formation of diacetylene (C4H2) is pinpointed using state-of-the art theoretical methods, accounting for high-order electron correlation, relativistic effects, non-Born-Oppenheimer corrections, and vibrational anharmonicity. Molecular energies are determined from coupled cluster theory with single and double excitations (CCSD), perturbative triples [CCSD(T)], full triples (CCSDT), and perturbative quadruples [CCSDT(Q)], in concert with correlation-consistent basis sets (cc-pVXZ, X=D, T, Q, 5, 6) that facilitate extrapolations to the complete basis set limit. The first full quartic force field of diacetylene is determined at the highly accurate all-electron CCSD(T) level with a cc-pCVQZ basis, which includes tight functions for core correlation. Application of second-order vibrational perturbation theory to our anharmonic force field yields fundamental frequencies with a mean absolute difference of only 3.9 cm(-1) relative to the experimental band origins, without the use of any empirical scale factors. By a focal point approach, we converge on an enthalpy change for the isogyric reaction 2 H-C[triple bond]C-H-->H-C[triple bond]C C[triple bond]C-H+H2 of (+0.03, +0.81) kcal mol(-1) at (0, 298.15) K. With the precisely established fHdegrees of acetylene, we thus obtain DeltafHdegrees(C4H2)=(109.4,109.7)+/-0.3 kcal mol(-1) at (0, 298.15) K. Previous estimates of the diacetylene enthalpy of formation range from 102 to 120 kcal mol(-1). PMID- 19191380 TI - Mode-specific photoionization dynamics of a simple asymmetric target: OCS. AB - Vibrationally resolved photoelectron spectra of OCS(+)(C (2)Sigma(+)) are used to probe coupling between photoelectron motion and molecular vibration for a simple asymmetric system. Spectra are reported over the photon energy range of 21Na(+)+I(-)) and the covalent channel (NaI-->Na+I). A Stark pulse is used to control the diabatic dissociation dynamics and a probe pulse is employed to ionize the products from the two channels. Based on the first order nonresonant nonperturbative dynamic Stark effect, the dissociation probabilities and the branching ratio of the products from the two channels can be controlled. Moreover the final photoelectron kinetic energy distribution can also be affected by the Stark pulse. The influences of the delay time, intensity, frequency, and carrier-envelope phase of the Stark pulse on the dissociation and ionization dynamics of the NaI molecule are discussed in detail. PMID- 19191387 TI - NMR shielding constants in BF(3) and magnetic dipole moments of (11)B and (10)B nuclei. AB - Gas-phase NMR spectra of (11)B, (10)B, and (19)F in BF(3) are reported, and high level ab initio calculations of the corresponding NMR shielding constants are described. Extrapolation of the measured resonance frequencies to the zero density limit ensures that the results correspond to the ab initio values for an isolated molecule. Simultaneous measurements of (3)He resonance frequencies and application of the calculated shielding constants allow us to determine improved values of the nuclear magnetic dipole moments of (11)B and (10)B. The magnetic moments of both isotopes are also determined independently by comparing with the (19)F spectral parameters (frequencies and shielding constants). The separately derived nuclear magnetic moments are in good agreement, whereas the literature moments of both (11)B and (10)B are noticeably less accurate. PMID- 19191388 TI - Photodissociation of the propargyl and propynyl (C(3)D(3)) radicals at 248 and 193 nm. AB - The photodissociation of perdeuterated propargyl (D(2)CCCD) and propynyl (D(3)CCC) radicals was investigated using fast beam photofragment translational spectroscopy. Radicals were produced from their respective anions by photodetachment at 540 and 450 nm (below and above the electron affinity of propynyl). The radicals were then photodissociated at 248 or 193 nm. The recoiling photofragments were detected in coincidence with a time- and position sensitive detector. Three channels were observed: D(2) loss, CD+C(2)D(2), and CD(3)+C(2). Observation of the D loss channel was incompatible with this experiment and was not attempted. Our translational energy distributions for D(2) loss peaked at nonzero translational energy, consistent with ground state dissociation over small (<1 eV) exit barriers with respect to separated products. Translational energy distributions for the two heavy channels peaked near zero kinetic energy, indicating dissociation on the ground state in the absence of exit barriers. PMID- 19191389 TI - Reading molecular messages from high-order harmonic spectra at different orientation angles. AB - We investigate the orientation dependence of high-order harmonic generation (HHG) from H(2) (+) with different internuclear distances irradiated by intense laser fields both numerically and analytically. The calculated molecular HHG spectra are found to be sensitive to the molecular axis orientation relative to incident laser field polarization and internuclear separation. In particular, our simulations demonstrate that at certain harmonic orders the envelopes of the HHG spectra taken at different orientation angles intersect. Moreover, the position of intersection is largely independent of the laser intensity while strongly dependent on the internuclear separation. This striking "intersection" phenomenon is identified as arising due to intramolecular two-center interference in the HHG and can be used to probe the molecular instantaneous structure. PMID- 19191390 TI - Theoretical investigation of the direct observation of anharmonic coupling in CDCl(3) in the time domain with femtosecond stimulated Raman scattering. AB - The femtosecond stimulated Raman scattering (FSRS) difference spectra of CDCl(3) from the ground potential energy surface, with and without off-resonance impulsive stimulated Raman pumping, taken at various delay times from the impulsive pump by Kukura et al. [Phys. Rev. Lett. 96, 238303 (2006)], showed sidebands of the C-Cl bends with periodic changes in phase as well as having an inversion symmetry to the high and low frequency sides of the Stokes C-D stretch at 2255 cm(-1). The semiclassical coupled-wave approach and a one-dimensional (1D) oscillator model for the C-D stretch whose frequency is modulated in time by the C-Cl bends could account for some features of the experimental results. Here, a quantum mechanical investigation is made of the FSRS difference spectra with a modulated 1D oscillator and three-dimensional (3D) harmonic and anharmonic potentials. It is shown that (i) the sidebands are allowed, with or without anharmonic coupling between the C-D stretch and the C-Cl bends, (ii) in the 3D harmonic model the sidebands have mirror symmetry about the Stokes C-D stretch, (iii) in the 3D anharmonic model with appropriate coupling terms between the C-D stretch and the C-Cl bends, the experimental results are well accounted for in both the phase changes in the FSRS difference spectra with time delay as well as the inversion symmetry of the sidebands for the C-Cl bends about the Stokes C-D stretch, (iv) there is a correspondence between the phase changes in the FSRS difference spectra and the wavepacket motion induced by the impulsive pump pulse on the ground potential energy surface as a function of the delay time, and (v) changing the polarization of the impulsive pump pulse, say, directly affects the coordinate dependence of the linear transition dipole moment in the asymmetric mode and hence its phase in the FSRS difference spectra, but not so for the symmetric modes where the displacement of the excited state surface governs the FSRS difference spectra more than the transition dipole moment. This work illustrates the potential of off-resonance FSRS in studying polyatomic molecular dynamics. PMID- 19191391 TI - Analytical potential energy surfaces for N(3) low-lying doublet states. AB - Adiabatic potential energy surfaces (PESs) for five low lying doublet states (three (2)A(') states and two (2)A(") states) of N(3) are constructed based on 1504 single point calculations at the MRCISD(Q) level using aug-cc-pVTZ basis set. A new strategy is adopted to obtain the final PESs by combining global fits of individual adiabatic PESs and local simultaneous fits of two adiabatic PESs in several conical intersection regions with switching functions. These global fits employ basis functions that satisfy permutational invariance with respect to like nuclei and have rms errors around 2-3 kcal/mol. The special local two-state fits are performed at the cyclic, bent, and linear N(3) conical intersection regions to take account of intrinsic square root behavior of the potentials and to improve the quality of fitting. Stationary points as well as minima on the concial intersections and seams of crossing are located on these PESs and compared with ab initio results. The agreement is satisfactory in most cases. In addition to the construction of adiabatic PESs, diabatization is performed for the 1 (2)A(') and 2 (2)A(') states around their conical intersection at the N(3) bent region. These two diabatic PESs and the diabatic coupling potential have been constructed and reported. PMID- 19191392 TI - Solvent effects on resonance Raman and hyper-Raman scatterings for a centrosymmetric distyrylbenzene and relationship to two-photon absorption. AB - Resonance Raman (RR) and resonance hyper-Raman (RHR) spectra have been obtained for a nominally centrosymmetric dialkylammonium-substituted distyrylbenzene that was previously shown to exhibit a strong solvent polarity effect on its two photon absorption strength. Spectra have been obtained in acetonitrile, methanol, dimethylsulfoxide, and water. In each solvent, the RR and RHR spectra show very similar intensity patterns in the strongest lines in both the C=C and C-C stretching regions, but the RHR peaks are shifted to higher energies by 5-10 cm( 1). These are interpreted as vibrations of similar mode character but different symmetries, with u symmetry modes observed in RHR and g symmetry modes in RR. In all solvents the nonzero chromophore contribution to the hyper-Rayleigh scattering indicates that the center of symmetry is broken to some degree, and this effect is more pronounced in water than in the other three solvents. The ratio of RR intensity to one-photon fluorescence intensity shows an approximate inverse relationship with the fluorescence quantum yield, consistent with the approximate solvent independence of the one-photon transition strength implied by the linear absorption spectra. The ratio of RHR intensity to two-photon fluorescence intensity is smaller than the corresponding RR to one-photon fluorescence ratio in all four solvents. PMID- 19191393 TI - Thermodynamics of ternary electrolytes: Enhanced adsorption of macroions as minority component to liquid interfaces. AB - We study the equilibrium thermodynamics between two ternary ionic systems in immiscible solvents characterized by different dielectric constants. We consider system geometries wherein the two phases of immiscible solvents occupy, respectively, semi-infinite regions of space separated by neutral and charged planar interfaces. Specifically we analyze the case where the ternary system is composed of a pair of symmetric ions plus a minority charged component of high valence. We describe the system by means of a nonlinear mean-field theory. We first obtain exact analytical solutions for the electrostatic potentials, as well as density profiles for a symmetric binary system, and then extend these results to the ternary case using the perturbation theory. We show that the corresponding adsorption and depletion of multivalent macroions at the interface are highly enhanced when compared with the monovalent case. PMID- 19191394 TI - Discovery of an intermediate phase in bis-thiourea pyridinium chloride inclusion compound. AB - This paper reports the occurrence of a new phase for bis-thiourea pyridinium chloride inclusion compound, which is intermediate to the high- and low temperature ones already reported. The structure of the intermediate phase was determined by the x-ray diffraction and the following sequence of space groups was obtained: Cmcm-->(237 K)Cmc2(1)-->(206 K)Pbca. The triplication of the lattice a parameter in the intermediate phase was observed. Detailed investigations of its phase transitions were performed by differential scanning calorimetry, x-ray diffraction, and dielectric spectroscopy methods. The phase transition at T(1)=237 K is continuous while at T(2)=206 K is discontinuous. Both phase transitions have been classified as of order-disorder type. The high- and low-temperature phases are nonpolar in contrast to the intermediate phase, for which we could not found ferroelectric ordering. PMID- 19191395 TI - Evolutionary algorithms to solve complicated NMR spectra. AB - The complexity of (1)H NMR spectra of solutes in partially ordered solvents such as liquid crystals increases rapidly with the number of spins. Spectra of simple solutes with sufficient symmetry and containing not too many spins (typically , where z is coordination number, is average cosine of angle between adjacent dipoles in liquids ([J. G. Kirkwood, J. Chem. Phys. 7, 911 (1939)], Eq. (19)) that Kirkwood factor as determined by the mean cosine of the angle between adjacent dipoles should be modified for the liquids consisting of nonlinear molecules with the complex type of the supramolecular organization. Alcohols are examples of such liquids, in which long-range intermolecular correlations beyond the nearest coordination shells are essential for their structural organization. PMID- 19191423 TI - A vapor cell based on dispensers for laser spectroscopy. AB - We describe a simple strontium vapor cell for laser spectroscopy experiments. Strontium vapor is produced using an electrically heated commercial dispenser source. The sealed cell operates at room temperature, and without a buffer gas or vacuum pump. The cell was characterized using laser spectroscopy, and was found to offer stable and robust operation, with an estimated lifetime of >10 000 h. By changing the dispenser, this technique can be readily extended to other alkali and alkaline earth elements. PMID- 19191424 TI - Automatic beam alignment system for a pulsed infrared laser. AB - A fully automatic alignment system for a pulsed infrared laser beam (5 ns pulses, 10 Hz repetition rate, 1.3 microm wavelength) was developed and tested. It compensates for long-term fluctuations of the beam initial position and direction the automatic realignment is performed every 10 min, and lasts typically 1-2 min. The system adjusts the beam initial position with a maximum error of 0.5 mm (10% of the beam diameter) and the beam direction with a maximum error of 50 murad. The solution is based on two InGaAs quadrant photodiodes as the position detectors and two motorized mirrors controlled by a personal computer. The signals from the quadrant detectors are processed by a peak detector and digitized by an analog to digital converter, which is synchronized with the laser pulses. PMID- 19191425 TI - Cryogenic ion trapping systems with surface-electrode traps. AB - We present two simple cryogenic rf ion trap systems in which cryogenic temperatures and ultra high vacuum pressures can be reached in as little as 12 h. The ion traps are operated either in a liquid helium bath cryostat or in a low vibration closed cycle cryostat. The fast turn around time and availability of buffer gas cooling made the systems ideal for testing surface-electrode ion traps. The vibration amplitude of the closed cycled cryostat was found to be below 106 nm. We evaluated the systems by loading surface-electrode ion traps with (88)Sr(+) ions using laser ablation, which is compatible with the cryogenic environment. Using Doppler cooling we observed small ion crystals in which optically resolved ions have a trapped lifetime over 2500 min. PMID- 19191426 TI - A modified commercial scanner as an image plate for table-top optical applications. AB - A reliable, accurate, and inexpensive optical detector for table-top applications is described here. Based on a commercial high resolution office scanner coupled to a projection plate, the detector offers a large image plate surface, allowing recording of sizeable images without systematic errors associated with coupling optics aberrations. Several tests on distance-dependent and steady interference patterns are presented and discussed. The extension to other types of optical measurements by substituting the projection plate is proposed. PMID- 19191427 TI - Advanced intensity correlation method for evaluating Poisson's ratio of fiberlike material. AB - An advanced intensity correlation method for evaluating Poisson's ratio of fiberlike materials is presented. The method is based on wave diffraction theory and digital image correlation (DIC) analysis. With the new method, the transverse strain of the fiberlike specimen under stretch can be conveniently and accurately determined by fitting the DIC-measured deformation distributions as a straight line following the theoretical diffraction model. The validity and flexibility of the method have been verified by numerical simulations and an experiment. The method can be readily extended to the characterizations of many fiberlike materials. PMID- 19191428 TI - Conceptual design of International Linear Collider damping ring wiggler based on a hybrid technology. AB - A magnetic design of a failure-free damping ring wiggler with 100% duty factor, 55.6 mm gap, and field exceeding 1.6 T is proposed. The insertion device is based on permanent magnets and specially shimmed poles that capable to meet the requirements of the International Linear Collider (ILC) damping rings (including positron one), with field quality and gap comparable to that projected for ILC using superconducting wiggler. Performance improvement of the modified hybrid design is attained due to yoke reduction down to six separated strips and maximized packaging of the midgrade (or high-grade) parallelepiped magnet blocks surrounding the Permendur poles. Economical efficiency is demonstrated on the base of calculated amount of magnetic material. A cryogenic variant of the hybrid design may certainly provide good sustainability to harsh radiation environment and further enhancement of the design efficiency within the state of the art. PMID- 19191429 TI - rf streak camera based ultrafast relativistic electron diffraction. AB - We theoretically and experimentally investigate the possibility of using a rf streak camera to time resolve in a single shot structural changes at the sub-100 fs time scale via relativistic electron diffraction. We experimentally tested this novel concept at the UCLA Pegasus rf photoinjector. Time-resolved diffraction patterns from thin Al foil are recorded. Averaging over 50 shots is required in order to get statistics sufficient to uncover a variation in time of the diffraction patterns. In the absence of an external pump laser, this is explained as due to the energy chirp on the beam out of the electron gun. With further improvements to the electron source, rf streak camera based ultrafast electron diffraction has the potential to yield truly single shot measurements of ultrafast processes. PMID- 19191430 TI - Influence of the electron cyclotron resonance plasma confinement on reducing the bremsstrahlung production of an electron cyclotron resonance ion source with metal-dielectric structures. AB - The influence of metal-dielectric (MD) layers (MD structures) inserted into the plasma chamber of an electron cyclotron resonance ion source (ECRIS) onto the production of electron bremsstrahlung radiation has been studied in a series of dedicated experiments at the 14 GHz ECRIS of the Institut fur Kernphysik der Universitat Frankfurt. The IKF-ECRIS was equipped with a MD liner, covering the inner walls of the plasma chamber, and a MD electrode, covering the plasma-facing side of the extraction electrode. On the basis of similar extracted currents of highly charged ions, significantly reduced yields of bremsstrahlung radiation for the "MD source" as compared to the standard (stainless steel) source have been measured and can be explained by the significantly better plasma confinement in a MD source as compared to an "all stainless steel" ECRIS. PMID- 19191431 TI - Experimental system to evaluate the effective diffusion coefficient of radon. AB - The effective diffusion coefficient of radon is a very important factor in estimating the rate of radon exhalation from the ground surface. In this study, we developed an experimental system that overcomes technical problems in previous studies to accurately evaluate the effective diffusion coefficient. The radon source used for this system was the National Institute of Radiological Sciences radon chamber. This chamber is a calibrated international standard facility that can produce stable radon concentrations for long periods of time. Our tests showed that leakage of radon from the system was negligible. After the leakage test, we evaluated the effective diffusion coefficient in free-space and in dry porous materials at porosities of 35% and 45%. To ensure that the porous material in the column was as homogeneous as possible, we filled the column with an artificial soil with controlled grain size and grain composition. The measured values and theoretical calculations agreed well, which indicate that the proposed system can be used to accurately and quickly evaluate the effective diffusion coefficient. PMID- 19191432 TI - Three point method to characterize low-pressure electronegative discharges using electrostatic probe. AB - Electrostatic probe measurements for low-pressure inductively coupled SF(6) plasmas are performed. From the current-voltage (I-V) curves of probe, the saturation currents of the positive ions and electrons and the electron temperature are measured. The electronegativity and the negative ion density are deduced by using the ratios of these parameters at three adjacent pressure points. The positive ion density is calculated by the orbital-motion-limited theory, and the electron temperatures are given either by the slope of the I-V curves or by the electron energy distribution function with the second derivative of I-V curves. The variations in the charged species density with pressure and power are investigated. PMID- 19191433 TI - Runaway electron energy measurement using hard x-ray spectroscopy in "Damavand" tokamak. AB - Set of experiments has been developed to study existing runaway electrons in "Damavand" tokamak plasma upon characteristics of hard x-ray emissions produced by collision of the runaway electrons with the plasma particles and limiters. As a first step, spatial distribution of hard x-ray emissions on the equatorial plane of the torus was considered. Obtained spectra of hard x-ray emissions for different alignments of shielded detector indicate isotropic emissivity in the equatorial plane. This is in agreement with wide angle cone of bremsstrahlung radiations, deduced from the mean value of energy of the runaway electrons. The mean energy was calculated from the slope of the energy spectrum of hard x-ray photons. In the second stage in order to investigate time evolution of energy of the runaway electrons, similar technique were applied to obtain hard x-ray energy in every 3 ms intervals, from the beginning to the end of plasma. The mean energy of the runaway electrons increases during the ramp up phase and reaches its maximum between 3 and 9 ms after plasma formation. Also considering the time dependence of the counted photons in each energy range shows that energetic photons are emitted during the ramp up phase of the plasma current in Damavand tokamak. PMID- 19191434 TI - Real-time feedback control of millimeter-wave polarization for LHD. AB - Electron cyclotron heating (ECH) is widely used in magnetic fusion devices, and the polarization of the injected millimeter-wave beams plays a crucial role in the propagation and absorption of the beam energy by the plasma. This polarization can be adjusted by grating mirror polarizers placed in the transmission lines which carry the microwaves from the power source to the plasma. In long-pulse devices such as the Large Helical Device (LHD) and ITER, it is desirable to track changes in the plasma and adjust the polarization of the ECH in real time such as to keep the absorption as high as possible and avoid shine-through which may lead to overheating of vessel components. For this purpose a real-time feedback control scheme is envisioned in which a measure of the absorption efficiency can be used to adjust the orientation of the polarizing mirrors toward an optimum. Such a setup has been tested in a low-power test stand as preparation for future implementation in the LHD ECH system. It is shown that a simple search algorithm is efficient and can in principle be used to control either the absorption efficiency or the linear polarization angle. PMID- 19191435 TI - Fourier-transform resonance shear measurement for studying confined liquids. AB - The resonance shear measurement we recently developed is an efficient tool for investigating the rheological and tribological properties of liquid nanofilms confined between solid surfaces with varying film thicknesses. However, the previously employed resonance shear measurement measured shear responses at various oscillation frequencies for one film thickness (the frequency scanning method), so it required at least several minutes to obtain a reliable resonance curve. The fast Fourier-transform resonance shear method was developed to rapidly acquire the resonance curves. The obtained curves were very similar to those obtained by the frequency scanning method. The time necessary for the FFT method was 2-10 s, much shorter than the time required for the scanning method. This technique is easy to use and extends the applicability of the method to volatile liquids and unstable samples. PMID- 19191436 TI - Model-free iterative control of repetitive dynamics for high-speed scanning in atomic force microscopy. AB - We introduce an algorithm for calculating, offline or in real time and with no explicit system characterization, the feedforward input required for repetitive motions of a system. The algorithm is based on the secant method of numerical analysis and gives accurate motion at frequencies limited only by the signal-to noise ratio and the actuator power and range. We illustrate the secant-solver algorithm on a stage used for atomic force microscopy. PMID- 19191437 TI - Design considerations for refractive solid immersion lens: application to subsurface integrated circuit fault localization using laser induced techniques. AB - With fast scaling and advancement of integrated circuit (IC) technology, circuitries have become smaller and denser. New materials and more sophisticated designs have evolved. These changes reduced the effectiveness of conventional laser induced fault localization techniques. Since IC fault localization is the most critical step in failure analysis, there are strong motivations to improve both spatial resolution and sensitivity of such systems to meet the new challenges from advanced technology. Refractive solid immersion lens (RSIL) is well known to enhance the laser spot size which directly affects resolution and sensitivity in back side fault localizations. In practice, it is difficult to operate RSIL at the ideal configurations to obtain the smallest spot resolution. It is necessary to understand the resolution performance at the other design focal planes. Besides resolution, there are also other factors that affect sensitivity in a RSIL enhanced system. This paper identifies and characterizes key RSIL design parameters to optimize RSIL performance on laser induced techniques. We report that the most efficient conditions are achieved close to aplanatic RSIL design to within 20-25 microm (for a 1 mm diameter lens), and the backing objective should be the minimum numerical aperture required for optimum resolution performance. The size of the mechanical clear aperture opening should be large enough (>80%) to exploit the advantage of aplanatic RSIL. RSIL is developed on a laser scanning optical microscope in this work, and a resolution of 0.3 microm (for a wavelength of 1340 nm) was achieved over a range of operating conditions. A quantitative resolution of 0.25 microm is achieved and a pitch structure of 0.4 microm is easily resolvable. Close to 15 times enhancement in laser induced signal is obtained. PMID- 19191438 TI - Low-temperature and high magnetic field dynamic scanning capacitance microscope. AB - We demonstrate a dynamic scanning capacitance microscope (DSCM) that operates at large bandwidths, cryogenic temperatures, and high magnetic fields. The setup is based on a noncontact atomic force microscope (AFM) with a quartz tuning fork sensor for the nonoptical excitation and readout in topography, force, and dissipation measurements. The metallic AFM tip forms part of a rf resonator with a transmission characteristics modulated by the sample properties and the tip sample capacitance. The tip motion gives rise to a modulation of the capacitance at the frequency of the AFM sensor and its harmonics, which can be recorded simultaneously with the AFM data. We use an intuitive model to describe and analyze the resonator transmission and show that for most experimental conditions it is proportional to the complex tip-sample conductance, which depends on both the tip-sample capacitance and the sample resistivity. We demonstrate the performance of the DSCM on metal disks buried under a polymer layer and we discuss images recorded on a two-dimensional electron gas in the quantum Hall effect regime, i.e. at cryogenic temperatures and in high magnetic fields, where we directly image the formation of compressible stripes at the physical edge of the sample. PMID- 19191439 TI - Laser scanning confocal microscope with programmable amplitude, phase, and polarization of the illumination beam. AB - We describe the design and construction of a laser scanning confocal microscope with programmable beam forming optics. The amplitude, phase, and polarization of the laser beam used in the microscope can be controlled in real time with the help of a liquid crystal spatial light modulator, acting as a computer generated hologram, in conjunction with a polarizing beam splitter and two right angled prisms assembly. Two scan mirrors, comprising an on-axis fast moving scan mirror for line scanning and an off-axis slow moving scan mirror for frame scanning, configured in a way to minimize the movement of the scanned beam over the pupil plane of the microscope objective, form the XY scan unit. The confocal system, that incorporates the programmable beam forming unit and the scan unit, has been implemented to image in both reflected and fluorescence light from the specimen. Efficiency of the system to programmably generate custom defined vector beams has been demonstrated by generating a bottle structured focal volume, which in fact is the overlap of two cross polarized beams, that can simultaneously improve both the lateral and axial resolutions if used as the de-excitation beam in a stimulated emission depletion confocal microscope. PMID- 19191440 TI - Multihit two-dimensional charged-particle imaging system with real-time image processing at 1000 frames/s. AB - A high-speed imaging system developed for two-dimensional counting of charged particles is presented. Microchannel plates coupled with a phosphor screen of a short emission lifetime (<1 micros) are used to visualize the two-dimensional positions of charged-particle impacts, and the image on the phosphor screen is captured with a 1 kHz complementary metal oxide semiconductor (CMOS) image sensor (512x512 pixels). A multistage image intensifier consisting of the first and second generation devices was used to compensate for the low sensitivity of CMOS. The centers of gravity (COG) of individual light spots in each image frame are calculated in real time by a field programmable gate array circuit. The performance of this system is tested by time-resolved photoelectron imaging (TR PEI) of NO using (1+1(')) resonance enhanced multiphoton ionization via the A (2)Sigma(+) state with a femtosecond laser operated at 1 kHz. The new system enabled COG detection for more than ten particles in each frame at 1 kHz and achieved an extremely high degree of accuracy in the measurement of photoelectron angular distributions in TR-PEI. PMID- 19191441 TI - The advanced ion-milling method for preparation of thin film using ion slicer: application to a sample recovered from diamond-anvil cell. AB - The advanced argon ion-milling technique using a new instrument called ion slicer was newly developed for preparation of thin foil. Compared to the conventional ion-milling methods, this technique facilitates very wide area to be homogeneously thinned by rocking the ion beam source with low angle and the specimen during milling. Here we applied this technique to a sample recovered from a laser-heated diamond-anvil cell (DAC). We obtained the thin film of almost entire cross section of the DAC sample along the compression axis, which possesses of approximately (10x50) microm(2). The laser-heated sample is often heterogeneous due to a large temperature gradient. However the chemical analyses are obtained from the whole hot spot under the transmission electron microscope by preparing the thin foil using the ion slicer. PMID- 19191442 TI - Measurement of femtosecond electron pulse length and the temporal broadening due to space charge. AB - The temporal width of ultrashort electron pulses as a function of beam intensity was measured on the femtosecond time scale with a customized streak camera. The results show that the temporal profile of an electron pulse is Gaussian at low beam intensity and progressively evolves to a top-hat shape due to space charge broadening as the beam intensity increases. The strong correlation between the pulse width and beam intensity observed in our streaking measurements agrees very well with the mean-field calculation and supports the main conclusion of previous theoretical studies that the space charge broadening plays a determinant role. PMID- 19191443 TI - A flyer-impact technique for measuring viscosity of metal under shock compression. AB - A flyer-impact technique, different from the explosive method of [Sakharov et al., Sov. Phys. Dokl. 9, 1091 (1965)], is developed to investigate the viscosity of shocked metals. The shock wave with a front of sinusoidal perturbation is induced by the sinusoidal profile of the impact surface of the sample by use of two-stage light-gas gun. The oscillatory damping process of the perturbation amplitude is monitored by electric pins. Two damping curves (perturbation amplitude relative to its initial value versus propagated distance relative to the wavelength of sinusoidal perturbation) of aluminum are determined at 78 and 101 GPa. The effective shear viscosity coefficients are deduced to be about 1300 and 800 Pa s based on the Miller and Ahrens analytic solution for viscous fluid. PMID- 19191444 TI - High-frequency electron spin resonance system using a microcantilever and a pulsed magnetic field. AB - A novel technique of high-frequency electron spin resonance (ESR) in a pulsed magnetic field is presented. Our technique is based on the magnetic detection of a magnetization change associated with the ESR absorption using a microcantilever. We successfully observed ESR signals of a microcrystal (mass approximately 1 microg) in the millimeter-wave region up to 130 GHz in pulsed magnetic fields of up to 2.4 T. This result corresponds to the spin sensitivity of approximately 10(11) spins/G, which is four orders of magnitude better than that of conventional transmission-type ESR techniques. PMID- 19191445 TI - An electrical microheater technique for high-pressure and high-temperature diamond anvil cell experiments. AB - Small electrical heating elements have been lithographically fabricated onto the culets of "designer" diamond anvils for the purpose of performing high-pressure and high-temperature experiments on metals. The thin-film geometry of the heating elements makes them very resistant to plastic deformation during high-pressure loading, and their small cross-sectional area enables them to be electrically heated to very high temperatures with relatively modest currents (approximately = 1 A). The technique also offers excellent control and temporal stability of the sample temperature. Test experiments on gold samples have been performed for pressures up to 21 GPa and temperatures of nearly 2000 K. PMID- 19191446 TI - Effects of the positioning force of electrostatic levitators on viscosity measurements. AB - Electrostatic levitators use strong electric fields to levitate and accurately position a sample against gravity. In this study, the effects of the electric field are investigated with regard to viscosity measurements conducted with the oscillating drop method. The effects of the external field on viscosity measurements are experimentally confirmed by changing the sample size. Moreover, a numerical simulation based on a simple mass-spring-damper system can reproduce the experimental observations. Based on the above results, measurement procedures are improved. These help to minimize the effect of the positioning force and to increase the accuracy of the viscosity measurements. PMID- 19191447 TI - Surface photovoltage spectroscopy in a Kelvin probe force microscope under ultrahigh vacuum. AB - Surface photovoltage (SPV) spectroscopy is a common method for optoelectronic semiconductor characterization. Kelvin probe force microscopy has developed into a widely used tool for nanoscale characterization of semiconductors, metals, and insulators. We present here a setup for the measurement of local SPV spectra in a Kelvin probe force microscope operated under ultrahigh vacuum conditions. The atomic force microscope tip can be placed to any desired position with nanometer precision and the SPV can then be recorded as a function of the wavelength of the illuminating light. We introduce the realization of the setup and present the SPV spectra on two test systems, an epitaxially grown GaAs/CuGaSe(2) junction and a Zn-doped CuInS(2) polycrystalline thin film. PMID- 19191448 TI - Development of a versatile high temperature top seeded solution growth unit for growing cesium lithium borate crystals. AB - The design and fabrication of a state of the art high temperature top seeded solution growth (HTTSSG) unit with seed and crucible rotation options is discussed. Crystals from solid solutions of high viscosity could be grown using the setup. The fabricated setup is used to grow high optical quality single crystals of cesium lithium borate with dimensions 50x40x40 mm(3) for frequency conversion of ir to UV laser through harmonic generation. PMID- 19191449 TI - How to track protists in three dimensions. AB - We present an apparatus optimized for tracking swimming micro-organisms in the size range of 10-1000 microm, in three dimensions (3Ds), far from surfaces, and with negligible background convective fluid motion. Charge coupled device cameras attached to two long working distance microscopes synchronously image the sample from two perpendicular directions, with narrow band dark-field or bright-field illumination chosen to avoid triggering a phototactic response. The images from the two cameras can be combined to yield 3D tracks of the organism. Using additional, highly directional broad-spectrum illumination with millisecond timing control the phototactic trajectories in 3D of organisms ranging from Chlamydomonas to Volvox can be studied in detail. Surface-mediated hydrodynamic interactions can also be investigated without convective interference. Minimal modifications to the apparatus allow for studies of chemotaxis and other taxes. PMID- 19191450 TI - Combining magnetic resonance imaging and ultrawideband radar: a new concept for multimodal biomedical imaging. AB - Due to the recent advances in ultrawideband (UWB) radar technologies, there has been widespread interest in the medical applications of this technology. We propose the multimodal combination of magnetic resonance (MR) and UWB radar for improved functional diagnosis and imaging. A demonstrator was established to prove the feasibility of the simultaneous acquisition of physiological events by magnetic resonance imaging and UWB radar. Furthermore, first in vivo experiments have been carried out, utilizing this new approach. Correlating the reconstructed UWB signals with physiological signatures acquired by simultaneous MR measurements, representing respiratory and myocardial displacements, gave encouraging results which can be improved by optimization of the MR data acquisition technique or the use of UWB antenna arrays to localize the motion in a focused area. PMID- 19191451 TI - Measurements of accurate x-ray scattering data of protein solutions using small stationary sample cells. AB - In this paper, we report a method of precise in situ x-ray scattering measurements on protein solutions using small stationary sample cells. Although reduction in the radiation damage induced by intense synchrotron radiation sources is indispensable for the correct interpretation of scattering data, there is still a lack of effective methods to overcome radiation-induced aggregation and extract scattering profiles free from chemical or structural damage. It is found that radiation-induced aggregation mainly begins on the surface of the sample cell and grows along the beam path; the diameter of the damaged region is comparable to the x-ray beam size. Radiation-induced aggregation can be effectively avoided by using a two-dimensional scan (2D mode), with an interval as small as 1.5 times the beam size, at low temperature (e.g., 4 degrees C). A radiation sensitive protein, bovine hemoglobin, was used to test the method. A standard deviation of less than 5% in the small angle region was observed from a series of nine spectra recorded in 2D mode, in contrast to the intensity variation seen using the conventional stationary technique, which can exceed 100%. Wide-angle x-ray scattering data were collected at a standard macromolecular diffraction station using the same data collection protocol and showed a good signal/noise ratio (better than the reported data on the same protein using a flow cell). The results indicate that this method is an effective approach for obtaining precise measurements of protein solution scattering. PMID- 19191452 TI - A lightweight balloon-carried cloud charge sensor. AB - Despite the importance of microphysical cloud processes on the climate system, some topics are under-explored. For example, few measurements of droplet charges in nonthunderstorm clouds exist. Balloon carried charge sensors can be used to provide new measurements. A charge sensor is described for use with meteorological balloons, which has been tested over a range of atmospheric temperatures from -60 to 20 degrees C, in cloudy and clear air. The rapid time response of the sensor (to >10 V s(-1)) permits charge densities from 100 fC m( 3) to 1 nC m(-3) to be determined, which is sufficient for it to act as a cloud edge charge detector at weakly charged horizontal cloud boundaries. PMID- 19191453 TI - An impact rotary motor based on a fiber torsional piezoelectric actuator. AB - A prototype small impact rotary motor has been fabricated based on a newly developed torsional actuator which is 15.0 mm long and 1.0 mm in diameter. The motor can rotate when it is powered with a saw-shaped voltage. The experimental results show that its angular speed is proportional to both the driving voltage's amplitude and the frequency under 1 kHz. The large nonlinearity occurs at higher driving frequency due to the resonance of the partial mechanical structure of the motor. The motor can rotate at a speed of 90 rpm with a saw-shaped driving voltage of 600 V(p.-p.) at 8 kHz, and produce a stall torque of 80 microN m with 1000 V(p.-p.) at 3 kHz. PMID- 19191454 TI - Laser ultrasonics and neural networks for the characterization of thin isotropic plates. AB - Nondestructive testing has been performed on a thin copper plate. Lamb waves were generated and studied with a noncontact method, using a laser ultrasonic setup. A two-dimensional Fourier transform technique has been employed to obtain the dispersion curves, revealing numerous symmetric and antisymmetric modes. The inverse problem, in other words, the determination of the thickness and the elastic constants of the tested plate, has been solved by means of a feed-forward neural network. These parameters were then evaluated simultaneously, the dispersion curves being entirely fitted. The experimental results show good agreement with the theoretical model. This inversion method was found to be prompt and easy to automate. PMID- 19191455 TI - Ripplon on high viscosity liquid. AB - A ripplon is a thermally excited capillary wave propagating on liquid surface. Propagation of the ripplon reflects surface mechanical properties such as surface tension and viscoelasticity of the liquid. An optical beating ripplon spectroscopy technique developed by us was applied mainly to the observation of various surface phenomena on a lowly viscous liquid surface in the frequency range from 1 kHz to several 10 MHz. In this study, we carried out a light scattering observation of the highly damped ripplon on the surface of the viscous liquid. The spectral peak width of the damped ripplon was smaller for the higher viscosity and the optical beating technique could resolve such a structure. A strict description of the dynamic structure factor of the ripplon was employed to fit the experimental power spectrum of overdamped ripplon to viscosity up to 1000 cS. PMID- 19191456 TI - Photoacoustic tomography imaging system based on digital B-mode ultrasound diagnosis equipment. AB - A photoacoustic tomography imaging system with B-mode digital ultrasound diagnosis equipment was designed, fabricated, and tested for biomedical imaging. The system for biomedical diagnostics was designed with four parts consisting of an irradiating source, digital B-mode ultrasound diagnostic equipment with a liner transducer array, data acquisition and transmission system, and rotation devices for imaging scanning. Multielement phase-controlled focus technique was applied to reduce the data-acquisition time and enhance signal-to-noise ratio. Under the self-designed working sequence, data-acquisition time of an image can be reduced to 12 s. A series of experiments were performed to estimate the imaging quality of the system. The lateral and elevational resolutions of the system were detected to be 0.25 and 1.4 mm, respectively. The results show that the system has the feasibility to be used as a medical imaging method, and it may provide a new rapid and noninvasive imaging modality for clinical applications. PMID- 19191457 TI - Chopper system for time resolved experiments with synchrotron radiation. AB - A chopper system for time resolved pump-probe experiments with x-ray beams from a synchrotron is described. The system has three parts: a water-cooled heatload chopper, a high-speed chopper, and a millisecond shutter. The chopper system, which is installed in beamline ID09B at the European Synchrotron Radiation Facility, provides short x-ray pulses for pump-probe experiments with ultrafast lasers. The chopper system can produce x-ray pulses as short as 200 ns in a continuous beam and repeat at frequencies from 0 to 3 kHz. For bunch filling patterns of the synchrotron with pulse separations greater than 100 ns, the high speed chopper can isolate single 100 ps x-ray pulses that are used for the highest time resolution. A new rotor in the high-speed chopper is presented with a single pulse (100 ps) and long pulse (10 micros) option. In white beam experiments, the heatload of the (noncooled) high-speed chopper is lowered by a heatload chopper, which absorbs 95% of the incoming power without affecting the pulses selected by the high speed chopper. PMID- 19191458 TI - Minimizing sample shape and size effects in all purpose magnetometers. AB - When measurements are made with a magnetometer, it is usually assumed that the sample produces a pure dipolar magnetic field. This is only true if the sample is a sphere. If the sample is not a sphere, the magnetic field produced by it will also contain higher order multipoles. These components of the magnetic field depend not only on the volume and magnetization but also on the sample's shape and size. Ideally the measured moment of a sample is only proportional to the sample's magnetization and volume. This paper explains why a sample's shape and size affects the response of a magnetometer, shows results illustrating this effect, and discusses conditions that will minimize these effects. PMID- 19191459 TI - Rotational UV lithography device for cylindrical substrate exposure. AB - Optical photolithograhy is a well developed technique, which is normally restricted to planar substrates used in microelectronics or microelectromechanical system fabrication. For other applications--e.g., patterning of stents--photolithography would be an attractive alternative to techniques such as laser structuring provided that the planar technique could be adapted to cylindrical geometries. This study presents the development of a three dimensional UV photolithography exposure method using a synchronizing movement between a planar Cr mask and a circular substrate. This technique was successfully applied to tubes with outer diameters between 1 and 5 mm. A lateral resolution for a 5 microm feature size of 4.8 microm was achieved, which is close to the resolution of 4.6 microm for similar planar films. PMID- 19191460 TI - Generation and control of thoron emanated from lantern mantles. AB - This paper describes the performance of a thoron ((220)Rn) flowthrough source made of a commercially available lantern mantle. This (220)Rn source is easy to construct and operate and has a negligible radon ((222)Rn) gas generated when the air was passed through the source. We studied the (220)Rn concentration generated from the lantern mantles in terms of the variability in concentration associated with both the total weight of the lantern mantles used in the source and the air flow rate used in the experimental chamber. We found that the concentration of (220)Rn generated in air ranged from 0.9 to 150 kBq m(-3) and exponentially depended on the absolute humidity. The (220)Rn concentration increased linearly with increasing total weight of the lantern mantles, but variations in the rate of air flow passed through the source had no influence on the observed (220)Rn concentration. PMID- 19191461 TI - A dynamic microindentation device with electrical contact detection. AB - We developed a microindentation instrument that allows direct measurement of the point of contact for reasonably conductive samples. This is achieved in the absence of a contact load using a simple electrical circuit. Force is measured using an optical interrupter to measure the deflection of a cantilever beam. Displacement is achieved using a piezoelectric motor and is measured using an independent optical interrupter. Force and displacement measurements are accomplished in real time, allowing the specification of arbitrary waveforms. The instrument was rigorously validated by comparing mechanical property measurements from the indenter with results from traditional dynamic mechanical analysis. Details of the construction and feedback control schemes are given explicitly. PMID- 19191462 TI - Precision optical trapping via a programmable direct-digital-synthesis-based controller for acousto-optic deflectors. AB - We describe a simple-to-construct programmable direct-digital-synthesis-based controller for use with acousto-optic deflectors. Our controller corrects for nonlinear diffraction efficiency versus diffraction angle, provides superior stability, functionality, and configurability, and costs a fraction of commercially available systems. Using this instrument, we move a 1 mum diameter bead by 1-nm-sized steps and resolve these steps. PMID- 19191463 TI - An automatic liquid-nitrogen filling system for multiple Ge detectors. AB - In order to reduce the time and effort of the daily task of refilling Dewar vessels of Ge detectors with liquid nitrogen (LN(2)), we have developed an automatic LN(2) filling system equipped with a LN(2) plant. With this system, we were freed from the work of LN(2) filling. Such an LN(2) filling system is useful for moderate-scale Ge detector arrays consisting of fewer than 20 Ge detectors. PMID- 19191464 TI - Compact collimated fiber optic array diagnostic for railgun plasmas. AB - We developed and tested a compact collimated 16 channel fiber optic array diagnostic for studying the light emission of railgun armature plasmas with approximately millimeter spatial and submicrosecond temporal resolution. The design and operational details of the diagnostic are described. Plasma velocities, oscillation, and dimension data from the diagnostic for the Livermore fixed hybrid armature experiment are presented and compared with one-dimensional simulations. The techniques and principles discussed allow the extension of the diagnostic to other railgun and related dense plasma experiments. PMID- 19191465 TI - Graphical template software for accurate micromanipulation in a scanning electron microscope. AB - Micromanipulation techniques in a scanning electron microscope (SEM) have been utilized for assembling micrometer-sized structures. The precision of the assembled microstructures has been limited by the poor accuracy of the SEM image. We have developed a software to assist the operator in the accurate assembly of microstructures in a SEM, in which computer-generated outlines of the target structure [graphical templates (GTs)] are superimposed on the monitor. The displayed GTs are distorted on the basis of the image properties of the SEM evaluated in advance. As a consequence, the operator can construct microstructures with a high precision only by maneuvering the manipulator so that the outline of each object perfectly overlaps the GT without any alteration of the electron optics or circuits for improving the image accuracy. PMID- 19191466 TI - A microfluidic Coulter counting device for metal wear detection in lubrication oil. AB - We present the design, fabrication, and testing of a microfluidic device for metal wear detection in lubrication oils. The detection is based on the capacitance Coulter counting principle, that is, on the change in a microchannel's capacitance caused by the presence of a metal particle in the microchannel. The testing of the microfluidic device using 10-25 microm aluminum particles has demonstrated the feasibility for detection and counting of microscale metal particles in low-conductive lubrication oils. This microfluidic device is promising for online oil debris detection by the use of multiple detection microfluidic channels. PMID- 19191467 TI - Design of high power radio frequency radial combiner for proton accelerator. AB - A simplified design method has been proposed for systematic design of novel radio frequency (rf) power combiner and divider, incorporating radial slab-line structure, without using isolation resistor and external tuning mechanism. Due to low insertion loss, high power capability, and rigid mechanical configuration, this structure is advantageous for modern solid state rf power source used for feeding rf energy to superconducting accelerating structures. Analysis, based on equivalent circuit and radial transmission line approximation, provides simple design formula for calculating combiner parameters. Based on this method, novel 8 way and 16-way power combiners, with power handling capability of 4 kW, have been designed, as part of high power solid state rf amplifier development. Detailed experiments showed good performance in accordance with theory. PMID- 19191468 TI - High power light emitting diode based setup for photobleaching fluorescent impurities. AB - Single molecule fluorescence experiments, with their associated low signals, require very low background fluorescence in the sample. Even high purity liquids will often possess large numbers of fluorescent impurities that are difficult to completely remove through standard purification techniques such as distillation and recrystallization. We have constructed a simple setup in which such impurities can be photobleached before final sample preparation. The instrument consists of high power light emitting diodes, and it delivers almost 10 W of light to the sample without the heating associated with more conventional light sources or the cost and safety concerns associated with a high power laser. PMID- 19191469 TI - Producing and quantifying enriched para-H2. AB - The production of enriched para-H(2) is useful for many scientific applications, but the technology for producing and measuring para-H(2) is not yet widespread. In this note and in the accompanying auxiliary material, we describe the design, construction, and use of a versatile standalone converter that is capable of producing para-H(2) enrichments of up to > or = 99.99% at continuous flow rates of up to 0.4 SLM. We also discuss para-H(2) storage and back conversion rates, and improvements to three techniques (thermal conductance, NMR, and solid hydrogen impurity spectroscopy) used to quantify the para-H(2) enrichment. PMID- 19191470 TI - Degradation and sorption of 2-propenyl and benzyl isothiocyanate in soil. AB - Isothiocyanates of natural origin produced by the hydrolysis of plant-produced glucosinolates have the potential to control soil pests, but getting sufficiently high isothiocyanate concentrations in soil is difficult. Furthermore, the isothiocyanates have proven toxic to a wide range of organisms and hence may also harm nontarget organisms. Knowledge of the sorption and degradation of the isothiocyanates is essential to optimize the use of natural isothiocyanates for pest control while minimizing the environmental impact. We have conducted studies on the sorption and degradation of two isothiocyanates of natural origin, 2 propenyl isothiocyanate and benzyl isothiocyanate. The experiments show the isothiocyanates degrade very quickly (t(1/2) = 0.93-4.25 h) in a 1:1 soil water slurry at 25 degrees C and they are sorbed by the organic matter in soil. From an environmental point of view, a fast degradation is desirable, but if the natural isothiocyanates are to be utilized for pest control, a fast degradation may imply they are not present long enough to have the desired effect on pests. PMID- 19191473 TI - Structure and insight into blue light-induced changes in the BlrP1 BLUF domain. AB - BLUF domains (sensors of blue light using flavin adenine dinucleotide) are a group of flavin-containing blue light photosensory domains from a variety of bacterial and algal proteins. While spectroscopic studies have indicated that these domains reorganize their interactions with an internally bound chromophore upon illumination, it remains unclear how these are converted into structural and functional changes. To address this, we have solved the solution structure of the BLUF domain from Klebsiella pneumoniae BlrP1, a light-activated c-di-guanosine 5' monophosphate phosphodiesterase which consists of a sensory BLUF and a catalytic EAL (Glu-Ala-Leu) domain [Schmidt et. al. (2008) J. Bacteriol. 187, 4774-4781]. Our dark state structure of the sensory domain shows that it adopts a standard BLUF domain fold followed by two C-terminal alpha helices which adopt a novel orientation with respect to the rest of the domain. Comparison of NMR spectra acquired under dark and light conditions suggests that residues throughout the BlrP1 BLUF domain undergo significant light-induced chemical shift changes, including sites clustered on the beta(4)beta(5) loop, beta(5) strand, and alpha(3)alpha(4) loop. Given that these changes were observed at several sites on the helical cap, over 15 A from chromophore, our data suggest a long-range signal transduction process in BLUF domains. PMID- 19191477 TI - Structure of a SusD homologue, BT1043, involved in mucin O-glycan utilization in a prominent human gut symbiont. AB - Mammalian distal gut bacteria have an expanded capacity to utilize glycans. In the absence of dietary sources, some species rely on host-derived mucosal glycans. The ability of Bacteroides thetaiotaomicron, a prominent human gut symbiont, to forage host glycans contributes to both its ability to persist within an individual host and its ability to be transmitted naturally to new hosts at birth. The molecular basis of host glycan recognition by this species is still unknown but likely occurs through an expanded suite of outermembrane glycan binding proteins that are the primary interface between B. thetaiotaomicron and its environment. Presented here is the atomic structure of the B. thetaiotaomicron protein BT1043, an outer membrane lipoprotein involved in host glycan metabolism. Despite a lack of detectable amino acid sequence similarity, BT1043 is a structural homologue of the B. thetaiotaomicron starch-binding protein SusD. Both structures are dominated by tetratrico peptide repeats that may facilitate association with outer membrane beta-barrel transporters required for glycan uptake. The structure of BT1043 complexed with N-acetyllactosamine reveals that recognition is mediated via hydrogen bonding interactions with the reducing end of beta-N-acetylglucosamine, suggesting a role in binding glycans liberated from the mucin polypeptide. This is in contrast to CBM 32 family members that target the terminal nonreducing galactose residue of mucin glycans. The highly articulated glycan-binding pocket of BT1043 suggests that binding of ligands to BT1043 relies more upon interactions with the composite sugar residues than upon overall ligand conformation as previously observed for SusD. The diversity in amino acid sequence level likely reflects early divergence from a common ancestor, while the unique and conserved alpha-helical fold the SusD family suggests a similar function in glycan uptake. PMID- 19191479 TI - Direct determination of the enantiomeric purity or enantiomeric composition of methylpropionates using a single capacitive microsensor. AB - Capacitive enantioselective sensors have been demonstrated to provide antipodal signals upon dosage of, e.g., the enantiomers of methyl lactate or methyl-2 chloropropionate. In a next step, these sensors have been used to not only qualitatively determine the nature of the respective enantiomer or to quantitatively measure its concentration upon dosage in the pure form but to also assess the enantiomeric composition of mixtures by using only a single capacitive type sensor. The enantioselective coating material consisted of a modified gamma cyclodextrin. It was shown that the absorption and desorption kinetics of the two enantiomers of, e.g., the methyl-2-chloropropionate, are sufficiently different and produce sensor signal features that enable an accurate determination of the enantiomeric purity and composition of the chiral analyte or mixture under investigation. The method even allows for detecting small impurities in commercially available samples labeled as 99% enantiomerically pure. Moreover, the results disclosed here show that sensor techniques can be used to reveal details of enantioselective analyte-receptor and analyte-matrix interactions. PMID- 19191484 TI - Comparison of QuantiFERON-TB Gold In-Tube Test and tuberculin skin test for identification of latent Mycobacterium tuberculosis infection in healthcare staff and association between positive test results and known risk factors for infection. AB - OBJECTIVE: We compared a whole-blood interferon-gamma release assay (QuantiFERON TB Gold In-Tube test, hereafter "QFT-in tube test") with a tuberculin skin test (TST) to determine which test more accurately identified latent Mycobacterium tuberculosis infection in healthcare staff. METHODS: A total of 481 hospital staff members were recruited from 5 hospitals in Melbourne, Australia. They provided information about demographic variables and tuberculosis (TB) risk factors (ie, birth or travel in a country with a high prevalence of TB, working in an occupation likely to involve contact with M. tuberculosis or individuals with TB, or being a household contact of an individual with a proven case of pulmonary TB). The QFT-in tube test and the TST were administered in accordance with standardized protocols. Concordance between the test results and positive risk factors was analyzed using the kappa statistic, the McNemar test, and logistic regression. RESULTS: A total of 358 participants had both a TST result and a QFT-in tube test result available for comparison. There were fewer positive QFT-in tube test results than positive TST results (6.7% vs. 33.0%; P<.001). Agreement between the tests was poor (71%; kappa=0.16). A positive QFT-in tube test result was associated with birth in a country with a high prevalence of TB, the number of years an individual had lived in a country with a high prevalence of TB (ie, the effect of each additional year, treated as a continuous variable), and high-risk occupational contact. A positive TST result was associated with older age, receipt of bacille Calmette-Guerin (BCG) vaccination, and working in an occupation that involved patient contact. Receipt of BCG vaccination was most strongly associated with discordant results in instances in which the TST result was positive and the QFT-in tube test result was negative. CONCLUSION: In a population of healthcare staff with a low prevalence of TB and a significant rate of BCG vaccination, a positive QFT-in tube test result was associated with the presence of known risk factors for TB exposure, whereas a positive TST result was more strongly associated with a prior history of BCG vaccination. PMID- 19191486 TI - In silico evaluation platform for artificial pancreatic beta-cell development--a dynamic simulator for closed-loop control with hardware-in-the-loop. AB - BACKGROUND: A critical step in algorithm development for an artificial beta-cell is extensive in silico testing. Computer simulations usually involve only the controller software, leaving untested the hardware elements, including the critical communication interface between the controller and the glucose sensor and insulin pump. METHODS: An in silico simulation platform has been developed that uses all of the components of the clinical system. At the core is a comprehensive in silico population model that covers the variability of principal metabolic parameters observed in vivo, to replace the human subject, with the ability to use historical clinical data. A continuous glucose monitor, in this case either the Abbott Diabetes Care (Alameda, CA) FreeStyle Navigator or the DexCom (San Diego, CA) STS7, is supplied with a glucose signal provided by the simulator. The Insulet (Bedford, MA) OmniPod insulin pump is also interfaced with the simulator to provide insulin delivery data. These hardware elements are an integral part of the system under testing, which also includes the algorithm components. RESULTS: The system is unique in that it uses the same hardware components for simulations as are required in clinical trials, allowing for full system level verification and validation. With a detailed mathematical model, a suite of patients can be simulated to reflect various conditions. Because all hardware is used, their related limitations are automatically included. CONCLUSIONS: A complete artificial beta-cell evaluation platform was realized with the flexibility to interface various algorithms and patient models, allowing for the systematic analysis of monitoring and control algorithms. The system facilitates a variety of tests and challenges to the software and the component devices, streamlining preclinical validation trials. PMID- 19191487 TI - A stable hyponitrite-bridged iron porphyrin complex. AB - The coupling of two nitric oxide (NO) molecules in heme active sites is an important contributor to the conversion of NO to nitrous oxide (N(2)O) by heme containing enzymes. Several formulations for the presumed heme-Fe{N(2)O(2)}(n-) intermediates have been proposed previously, however, no crystal structures of heme-Fe{N(2)O(2)}(n-) systems have been reported to date. We report the first isolation and characterization of a stable bimetallic hyponitrite iron porphyrin, [(OEP)Fe](2)(mu-N(2)O(2)), prepared from the reaction of [(OEP)Fe](2)(mu-O) with hyponitrous acid. Density functional theoretical calculations were performed on the model compound [(porphine)Fe](2)(mu-N(2)O(2)) to characterize its electronic structure and properties. PMID- 19191488 TI - Cross-resistance to herbicides of five ALS-inhibiting groups and sequencing of the ALS gene in Cyperus difformis L. AB - Resistance to ALS-inhibiting herbicides in Cyperus difformis has evolved rapidly in many rice areas worldwide. This study identified the mechanism of resistance, assessed cross-resistance patterns to all five chemical groups of ALS-inhibiting herbicides in four C. difformis biotypes, and attempted to sequence the ALS gene. Whole-plant and ALS enzyme activity dose-response assays indicated that the WA biotype was resistant to all ALS-inhibiting herbicides evaluated. The IR biotype was resistant to bensulfuron-methyl, orthosulfamuron, imazethapyr, and propoxycarbazone-sodium and less resistant to bispyribac-sodium and halosulfuron methyl, and susceptible to penoxsulam. ALS enzyme activity assays indicated that resistance is due to an altered target site yet mutations previously found to endow target-site resistance in weeds were not detected in the sequences obtained. The inability to detect resistance mutations in C. difformis may result from the presence of additional ALS genes, which were not amplified by the primers used. This study reports the first ALS gene sequence from Cyperus difformis. Certain ALS-inhibiting herbicides can still be used to control some resistant C. difformis biotypes. However, because cross-resistance to all five classes of ALS-inhibitors was detected in other resistant biotypes, these herbicides should only be used within an integrated weed management program designed to delay the evolution of herbicide resistance. PMID- 19191489 TI - 2-Aminopurine-modified abasic-site-containing duplex DNA for highly selective detection of theophylline. AB - 2-Aminopurine-modified abasic-site-containing duplex [DNA 5'-TCTGC GTCCT PXT TAACG CACAC-3'/3'-AGACG CAGGA TCA ATTGC GTGTG-5'; P = 2-aminopurine, X = abasic site (Spacer-C3), C = receptor base] is capable of selectively binding to the bronchodilator theophylline with a dissociation constant of 10 microM (5 degrees C, pH 7.0, I = 0.11 M) and is applicable to monitoring serum theophylline concentrations. PMID- 19191490 TI - Melting dramatically enhances the reactivity of aluminum nanoclusters. AB - The kinetic energy threshold for chemisorption of N(2) on Al(100)(+) has been measured as a function of the nanocluster's temperature from 440 to 790 K. When the Al(100)(+) cluster melts at 620-660 K, the threshold drops by approximately 1 eV (approximately 96 kJ/mol). A decrease in the activation energy of this magnitude causes a 10(8)-fold increase in the reaction rate at the melting temperature. The decrease in the activation energy may result from the mobility of the surface atoms on the liquid cluster, which allows them to move to a lower energy arrangement as the N(2) approaches. PMID- 19191491 TI - Ultrafast charge separation in multiexcited CdSe quantum dots mediated by adsorbed electron acceptors. AB - Ultrafast ET with a characteristic time constant of approximately 70 fs between CdSe QDs (mean radii of 1.4 nm) photoexcited in the lowest 1S electron state (lambda(exc) = 539 nm), and the molecular electron acceptor MV(2+) adsorbed on the QD surface was observed. The photophysics of such a system was investigated by time-resolved transient absorbance spectroscopy in the UV-visible spectral region. Our studies for the coupled system as a function of excitation intensity at lambda(exc) = 387 nm show that the ET processes compete efficiently with Auger recombination in CdSe QDs and at least 4 e-h pairs can be separated by ET to the electron acceptor MV(2+). PMID- 19191492 TI - Phthalocyanine dimerization-based molecular beacons using near-IR fluorescence. AB - Herein we demonstrate the use of a novel dimerization-based molecular beacon (MB) probe consisting of two metallo-phthalocyanine (Pc) fluorophores that use near-IR fluorescence, appropriate for highly specific and sensitive in vivo and/or in vitro DNA/RNA detection. Pc's possess a propensity to form nonfluorescent H dimers that is utilized as the molecular "off" switch in the closed MB conformation. The "on" switch, which is generated when the solution target binds to the loop of the MB forming the open form, also provides two fluorophores for transduction resulting in a doubling of the extinction coefficient and improving the resulting fluorescence yield compared to a classical single fluorophore/quencher MB system. In addition, the Pc-based MBs possess high thermal, photo, and chemical stabilities that are essential for many highly sensitive applications, such as molecular imaging. The dimer-based MBs were obtained using a simple single-step synthesis procedure and demonstrated excellent quenching efficiencies (98%) as well as a high signal-to-background ratio (approximately 60) exceeding the performance characteristics of many conventionally available MB probes. PMID- 19191493 TI - Millisecond curing time of a molecular adhesive causes velocity-dependent cargo loading of molecular shuttles. AB - It has been recently discovered that biological systems exploit complex intermolecular bonds, such as the "catch bond" between FimH proteins and mannose, to regulate dynamic assembly processes. Here we show that the assembly of hybrid nanostructures also has to account for and ideally exploit the subtleties of the utilized intermolecular interactions. We focus on the biotin-streptavidin bond, which gains its ultimate strength on a time scale of milliseconds, due to existence of metastable binding states. As a consequence of the gluelike character of this widely used intermolecular bond, the velocity of molecular shuttles, active nanoscale transport systems based on biomolecular motors and their associated filaments, has to be optimized to permit efficient attachment of cargo via biotin-streptavidin linkages. The attachment process can only be understood by combining rigorous mechanical engineering analysis with detailed physicochemical models. PMID- 19191494 TI - Effect of the chemical functionalization on charge transport in carbon nanotubes at the mesoscopic scale. AB - We present first-principles calculations of quantum transport in chemically functionalized metallic carbon nanotubes with lengths reaching the micrometer scale and random distributions of functional groups. Two typical cases are investigated, namely, a sp2-type bonding between carbene groups (CH2) and the nanotube sidewalls and a sp3-type bonding of nanotubes with paired phenyl groups. For similar molecular coverage density, charge transport is found to range from a quasi-ballistic-like to a strongly diffusive regime, with corresponding mean free paths changing by orders of magnitude depending on the nature of the chemical bonding. PMID- 19191495 TI - Osteocalcin secretion as an early marker of in vitro osteogenic differentiation of rat mesenchymal stem cells. AB - Osteocalcin (OC) is a bone-specific protein synthesized by osteoblasts that represents a good marker for osteogenic maturation. We examined whether in vitro osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells (MSCs) could be simply assessed at earlier stages by monitoring OC secretion into the conditioned medium, rather than measuring OC deposition on the extracellular matrix (ECM), using a sandwich enzyme immunoassay system involving a specific anti-rat OC monoclonal antibody. During a 16-day culture, OC was secreted into the medium of MSCs from day 8 and increased substantially until day 16. In contrast, OC deposition on the ECM was low, even at day 13, when calcium deposition was at high levels. The histological expression pattern of OC messenger RNA provided in situ evidence that osteoblastic cells appeared at the early stages of 6 to 9 days and matured over time in vitro. Furthermore, the temporal expression of osteogenesis-specific genes, such as the transcriptional factors core-binding factor 1 and osterix, followed by increases in secretory OC proved the commitment of MSCs to osteoblastic differentiation. These results revealed that biomineralization followed secretion of OC, which may reflect early osteoblastic differentiation of cultured MSCs under osteoinductive conditions. We ascertained the osteogenic differentiation capacity of cultured MSCs in a non destructive manner by monitoring OC secretion into the culture medium and proved that secretory OC could represent a reliable marker for predicting in vivo osteogenic potential in bone tissue engineering. PMID- 19191497 TI - High electron mobility in vacuum and ambient for PDIF-CN2 single-crystal transistors. AB - Single-crystal field-effect transistors (FETs) based on a fluorocarbon substituted dicyanoperylene-3,4:9,10-bis(dicarboximide) [PDIF-CN(2)] were fabricated by lamination of the semiconductor crystal on Si-SiO(2)/PMMA-Au gate dielectric-contact substrates. These devices were characterized both in vacuum and in the air, and they exhibit electron mobilities of ca. 6-3 and ca. 3-1 cm(2) V(-1) s(-1), respectively, I(on):I(off) > 10(3), and near-zero threshold voltage. PMID- 19191499 TI - Bone regeneration with autologous plasma, bone marrow stromal cells, and porous beta-tricalcium phosphate in nonhuman primates. AB - To potentiate the bone formation capability of bone marrow stromal cell (BMSC)/beta-tricalcium phosphate (beta-TCP) constructs, we devised an autologous plasma-based construct. We tested its effectiveness and investigated the effects of its components on a monkey ectopic bone formation model. The autologous plasma (platelet-rich plasma, PRP, or platelet-poor plasma, PPP)/BMSC/beta-TCP construct (R group or P group) showed significantly more bone formation at 3 and 6 weeks after implantation than a conventional BMSC/beta-TCP construct using a culture medium (M group). There was no significant difference between the P and R groups. Moreover, the P group constructs with a 10-fold lower cell concentration yielded equivalent bone formation to the M group at 5 weeks after implantation. To elucidate the effect of fibrin and serum contained in the plasma, five constructs were prepared using the following cell vehicles: autologous serum + fibrinogen (0, 1, 4, or 16 mg/mL) or phosphate-buffered saline + fibrinogen (4 mg/mL). The serum + fibrinogen (4 mg/mL, physiological concentration of monkeys) construct showed the most abundant bone formation at 3 weeks after implantation, though at 5 weeks no statistical difference existed among the groups. Autologous plasma efficiently promoted osteogenesis of BMSCs/porous beta-TCP constructs, and both fibrin and serum proved to play significant roles in the mechanism. PMID- 19191500 TI - Carbon nanotubes as molecular transporters for walled plant cells. AB - We have investigated the capability of single-walled carbon nanotubes (SWNTs) to penetrate the cell wall and cell membrane of intact plant cells. Confocal fluorescence images revealed the cellular uptake of both SWNT/fluorescein isothiocyanate and SWNT/DNA conjugates, demonstrating that SWNTs also hold great promise as nanotransporters for walled plant cells. Moreover, the result suggested that SWNTs could deliver different cargoes into different plant cell organelles. PMID- 19191498 TI - anti-Diastereo- and enantioselective carbonyl crotylation from the alcohol or aldehyde oxidation level employing a cyclometallated iridium catalyst: alpha methyl allyl acetate as a surrogate to preformed crotylmetal reagents. AB - Under the conditions of transfer hydrogenation employing an ortho-cyclometallated iridium catalyst generated in situ from [Ir(cod)Cl](2), 4-cyano-3-nitrobenzoic acid and the chiral phosphine ligand (S)-SEGPHOS, alpha-methyl allyl acetate couples to alcohols 1a-1j with complete levels of branched regioselectivity to furnish products of carbonyl crotylation 3a-3j, which are formed with good levels of anti-diastereoselectivity and exceptional levels of enantioselectivity. An identical set of optically enriched carbonyl crotylation products 3a-3j is accessible from the corresponding aldehydes 2a-2j under the same conditions, but employing isopropanol as the terminal reductant. Experiments aimed at probing the origins of stereoselection establish a matched mode of ionization for the (R) acetate and the iridium catalyst modified by (S)-SEGPHOS, as well as reversible ionization of the allylic acetate with rapid pi-facial interconversion of the resulting pi-crotyl intermediate in advance of C-C bond formation. Additionally, rapid alcohol-aldehyde redox equilibration in advance of carbonyl addition is demonstrated. Thus, anti-diastereo- and enantioselective carbonyl crotylation from the alcohol or aldehyde oxidation level is achieved in the absence of any stoichiometric metallic reagents or stoichiometric metallic byproducts. PMID- 19191501 TI - Combined effects of scaffold stiffening and mechanical preconditioning cycles on construct biomechanics, gene expression, and tendon repair biomechanics. AB - Our group has previously reported that in vitro mechanical stimulation of tissue engineered tendon constructs significantly increases both construct stiffness and the biomechanical properties of the repair tissue after surgery. When optimized using response surface methodology, our results indicate that a mechanical stimulus with three components (2.4% strain, 3000 cycles/day, and one cycle repetition) produced the highest in vitro linear stiffness. Such positive correlations between construct and repair stiffness after surgery suggest that enhancing structural stiffness before surgery could not only accelerate repair stiffness but also prevent premature failures in culture due to poor mechanical integrity. In this study, we examined the combined effects of scaffold crosslinking and subsequent mechanical stimulation on construct mechanics and biology. Autologous tissue-engineered constructs were created by seeding mesenchymal stem cells (MSCs) from 15 New Zealand white rabbits on type I collagen sponges that had undergone additional dehydrothermal crosslinking (termed ADHT in this manuscript). Both constructs from each rabbit were mechanically stimulated for 8h/day for 12 consecutive days with half receiving 100 cycles/day and the other half receiving 3000 cycles/day. These paired MSC collagen autologous constructs were then implanted in bilateral full-thickness, full-length defects in the central third of rabbit patellar tendons. Increasing the number of in vitro cycles/day delivered to the ADHT constructs in culture produced no differences in stiffness or gene expression and no changes in biomechanical properties or histology 12 weeks after surgery. Compared to MSC based repairs from a previous study that received no additional treatment in culture, ADHT crosslinking of the scaffolds actually lowered the 12-week repair stiffness. Thus, while ADHT crosslinking may initially stiffen a construct in culture, this specific treatment also appears to mask any benefits of stimulation among repairs postsurgery. Our findings emphasize the importance of properly preconditioning a scaffold to better control/modulate MSC differentiation in vitro and to further enhance repair outcome in vivo. PMID- 19191502 TI - Vortex polarization states in nanoscale ferroelectric arrays. AB - Two-dimensional arrays of ferroelectric lead zirconate titanate (PZT) nanodots were fabricated using pulsed laser deposition through ultrathin anodic aluminum oxide membrane stencil masks. The static distribution of polarization configurations was investigated using in- and out-of-plane piezoresponse force microscopy (PFM). The observed presence of an in-plane polarization component in nominally (001) oriented PZT suggests the existence of a significant deviation from the regular tetragonal structure that allows the formation of complex core polarization states. Core-polarization states may indicate the presence of quasi toroidal polarization ordering. The experimental results are compared with a theoretical model to determine the fingerprint of a vortex polarization state in PFM. PMID- 19191504 TI - Thermodynamic properties of aqueous gadolinium perrhenate and gadolinium chloride from high dilution calorimetry at extreme temperatures and pressures. AB - The heat of solution of solid cubic gadolinium oxide has been measured in noncomplexing perrhenic acid solutions at very high dilutions (10(-4) m) up to 596.30 K, from which the standard state thermodynamic properties of aqueous gadolinium perrhenate were determined up to 623.15 K. From the measured differences between similar properties of aqueous sodium chloride and perrhenate, thermodynamic properties for aqueous gadolinium chloride were obtained by ionic additivity. Data for the hydrolysis of Gd3+(aq) were obtained by separate determinations. The enthalpy of solution of gadolinium chloride at 623.15 K obtained from this research (-2.7 MJ mol(-1)) is apparently larger than any other recorded for a chemical reaction involving aqueous systems. Standard state partial molal heat capacities for ThCl4(aq) were predicted up to 623.15 K. PMID- 19191505 TI - Standard state thermodynamic properties of Ba2+(aq), Co2+(aq), and Cu2+(aq) up to 598.15 K, and temperature effect on ligand field. AB - Integral heat of solution measurements of barium chloride to 619.81 K, copper oxide in an excess of perrhenic acid to 585 K, and cobalt perrhenate in perrhenic acid to 573 K were measured in a high dilution calorimeter (< or =10(-3) m) at psat, from which the high temperature thermodynamic properties of aqueous barium chloride, copper perrhenate, and cobalt perrhenate were obtained. From the known differences between the corresponding properties for aqueous perrhenate and chloride ions, the thermodynamic properties of completely ionized aqueous copper and cobalt chloride were obtained from ionic additivity. The enthalpy and derived heat capacity data at higher temperatures (T > 473.15 K) suggest that the ligand field stabilization energy of Co2+(aq) may be disappearing. PMID- 19191503 TI - Phospholamban modulates the functional coupling between nucleotide domains in Ca ATPase oligomeric complexes in cardiac sarcoplasmic reticulum. AB - Oligomeric interactions between Ca-ATPase polypeptide chains and their modulation by phospholamban (PLB) were measured in native cardiac sarcoplasmic reticulum (SR) microsomes. Progressive modification of Lys(514) with fluorescein 5 isothiocyanate (FITC), which physically blocks access to the nucleotide binding site by ATP, demonstrates that Ca-ATPase active sites function independently of one another prior to the phosphorylation of PLB. However, upon cAMP-dependent protein kinase (PKA) phosphorylation of PLB, a second-order dependence between residual enzyme activity and the fraction of active sites is observed, consistent with a dimeric functional complex. Complementary distance measurements were made using FITC or 5-iodoacetamidofluorescein (IAF) bound to Cys(674) within the N- or P-domains, respectively, to detect structural coupling within oligomeric complexes. Accompanying the phosphorylation of PLB, neighboring Ca-ATPase polypeptide chains exhibit a 4 +/- 2 A decrease in the proximity between FITC sites within the N-domain and a 9 +/- 3 A increase in the proximity between IAF sites within P-domains. Thus, the phosphorylation of PLB induces spatial rearrangements between the N- and P-domain elements of proximal Ca-ATPase polypeptide chains which restore functional interactions between neighboring polypeptide chains and, in turn, result in increased rates of catalytic turnover. These results are interpreted in terms of a structural model, calculated through optimization of shape complementarity, desolvation, and electrostatic energies, which suggests a dimeric arrangement of Ca-ATPase polypeptide chains through the proximal association of N-domains that accommodates interaction with PLB. We suggest that the phosphorylation of PLB acts to release constraints involving interdomain subunit interactions that enhance catalytically important N-domain motions. PMID- 19191506 TI - A unified theory of the thermodynamic properties of aqueous electrolytes to extreme temperatures and pressures. AB - A new theoretical treatment has been developed for predicting the thermodynamic properties of electrolytes up to and beyond the critical temperature of water (973 K and at pressures up to 1000 MPa). The model is based upon the classical Born equation corrected for non-Born hydration effects. The temperature and pressure behavior of electrolytes can now be accurately predicted from existing low temperature data. Only two constants are needed for each electrolyte at all temperatures and pressures, where data exist to test the theory. PMID- 19191507 TI - Palladium(II)-catalyzed conjugate phosphination of electron-deficient acceptors. AB - A general protocol for the conjugate transfer of diphenyl-, dicyclohexyl-, and di tert-butylphosphinyl groups from silylphosphines to cyclic and acyclic electron deficient acceptors employing a bench-stable palladium(II) catalyst is reported. Several E and Z configured alpha,beta-unsaturated carbonyl and carboxyl acceptors (including imides) as well as nitroalkenes participate in this palladium(II) catalyzed process in high chemical yields. PMID- 19191508 TI - Electron transfer in oligothiophene-bridged bisporphyrins. AB - Oligothiophene-bridged zinc-tin bisporphyrinates were synthesized. Their absorption spectra have been analyzed in terms of exciton interaction and porphyrin-bridge coupling by through-bond interaction and the steady-state fluorescence spectra in terms of differential Stokes shifts for the electron donating zinc and the electron-accepting tin porphyrinates. Strong quenching of the fluorescence intensity and acceleration of the fluorescence decay as compared to porphyrinate monomers (ZnTPP, SnTPP) were observed. Both phenomena were traced back to light-induced electron transfer by the occurrence of ion pair absorption bands in picosecond transient absorption spectra. Similar absorption spectra of both chromophores caused always simultaneous excitation and, consequently, two concurrent photoreactions. Combined evaluation of the time-dependent absorption and fluorescence data allowed the estimation of rates for the electron transfer reactions. The found dependence on the separation distance was much smaller than for donor-acceptor systems with saturated spacers. A damping factor of 0.05 was calculated for the charge separation proceeding from the excited state of the zinc porphyrin. The polarity of the solvent had a profound influence on the transfer rates. The charge recombination was 300 times faster in polar tetrahydrofuran than in nonpolar toluene. PMID- 19191509 TI - Determination of the structure form of the fourth ligand of zinc in Acutolysin A using combined quantum mechanical and molecular mechanical simulation. AB - Acutolysin A, which is isolated from the snake venom of Agkistrodon acutus, is a member of the SVMPs subfamily of the metzincin family, and it is a snake venom zinc metalloproteinase possessing only one catalytic domain. The catalytic zinc ion, in the active site, is coordinated in a tetrahedral manner with three imidazole nitrogen atoms of histidine and one oxygen atom. It is uncertain whether this oxygen atom is a water molecule or a hydroxide ion just from the three-dimensional X-ray crystal structure. The identity of the fourth ligand of zinc is theoretically determined for the first time by performing both combined quantum mechanical and molecular mechanical (QM/MM) simulation and high-level quantum mechanical calculations. All of the results obtained indicate that the fourth ligand in the active site of the reported X-ray crystal structure is a water molecule rather than a hydroxide anion. On the basis of these theoretical results, we note that the experimental observed pH dependence of the proteolytic and hemorrhagic activity of Acutolysin A can be attributed to the deprotonation of the zinc-bound water to yield a better nucleophile, the hydroxide ion. Structural analyses revealed structural details useful for the understanding of acutolysin catalytic mechanism. PMID- 19191510 TI - Effects of sugars on lipid bilayers during dehydration--SAXS/WAXS measurements and quantitative model. AB - We present an X-ray scattering study of the effects of dehydration on the bilayer and chain-chain repeat spacings of dipalmitoylphosphatidylcholine bilayers in the presence of sugars. The presence of sugars has no effect on the average spacing between the phospholipid chains in either the fluid or gel phase. Using this finding, we establish that for low sugar concentrations only a small amount of sugar exclusion occurs. Under these conditions, the effects of sugars on the membrane transition temperatures can be explained quantitatively by the reduction in hydration repulsion between bilayers due to the presence of the sugars. Specific bonding of sugars to lipid headgroups is not required to explain this effect. PMID- 19191511 TI - Re-evaluation of the pressure effect for nucleation in laminar flow diffusion chamber experiments with fluent and the fine particle model. AB - This study is an investigation of the effect of total pressure on homogeneous nucleation rates of n-butanol in helium and n-pentanol in helium and argon in a laminar flow diffusion chamber (LFDC). To verify earlier findings, experimental data was re-evaluated using the computational fluid dynamics (CFD) software FLUENT in combination with the fine particle model (FPM) for aerosol dynamics calculations. This approach has been introduced in an earlier paper [Herrmann, E.; Lihavainen, H.; Hyvarinen, A.-P.; Riipinen, I.; Wilck, M.; Stratmann, F.; Kulmala, M. J. Phys. Chem. A 2006, 110, 12448]. As a result of our evaluation, a flaw in the femtube2 code was found which had been used in the original data analysis [Hyvarinen, A.-P.; Brus, D.; Zdimal, V.; Smolik, J.; Kulmala, M.; Viisanen, Y.; Lihavainen, H. J. Chem. Phys. 2006, 124, 224304]. The FLUENT analysis yielded a weak positive pressure effect for the nucleation of n-butanol in helium at low nucleation temperatures (265-270 K). n-Pentanol in helium showed a positive pressure effect at all temperatures (265-290 K), while the effect for the nucleation of n-pentanol in argon was negative at high temperatures (280 and 285 K) and positive at lower nucleation temperatures (265 K). These findings support results gained with the corrected femtube2 model. In this study, we also carried out a detailed comparison of FLUENT and femtube2 modeling results, especially focusing on the calculation of temperature and saturation ratio at nucleation rate maximum (T(nuc) and S(nuc), respectively) in both models. PMID- 19191512 TI - Electron transfer kinetics of cytochrome c probed by time-resolved surface enhanced resonance Raman spectroscopy. AB - An improved setup including a measuring cell was designed for time-resolved surface-enhanced resonance Raman (SERR) spectroscopy. The cell is based on a rotating disk electrode (RDE) made from electrochemically roughened Ag. Cytochrome c (cc) adsorbed on a monolayer of mercaptoethanol is investigated with respect to heterogeneous electron transfer. Cyclic voltammograms and potential dependent static SERR spectra indicate cc to be electroactive on the Ag electrode. The standard redox potential was found to be 234 mV. Time-resolved SERR spectra were then measured triggered by periodic potential pulses changing the protein between the oxidized and reduced state at a frequency of 10 Hz. Monoexponential functions obtained from the intensity of the band at 1361 cm-1 plotted versus time yielded the rate constants of heterogeneous electron transfer to be k(ox) = 46 +/- 7 s(-1) and k(red) = 84 +/- 20 s(-1). These relatively low rates are in line with the orientation of cc on the mercaptoethanol-modified Ag electrode. In this case the heme cleft pointed away from the surface thus hampering electron transfer. PMID- 19191513 TI - Neural stem cell- and Schwann cell-loaded biodegradable polymer scaffolds support axonal regeneration in the transected spinal cord. AB - Biodegradable polymer scaffolds provide an excellent approach to quantifying critical factors necessary for restoration of function after a transection spinal cord injury. Neural stem cells (NSCs) and Schwann cells (SCs) support axonal regeneration. This study examines the compatibility of NSCs and SCs with the poly lactic-co-glycolic acid polymer scaffold and quantitatively assesses their potential to promote regeneration after a spinal cord transection injury in rats. NSCs were cultured as neurospheres and characterized by immunostaining for nestin (NSCs), glial fibrillary acidic protein (GFAP) (astrocytes), betaIII-tubulin (immature neurons), oligodendrocyte-4 (immature oligodendrocytes), and myelin oligodendrocyte (mature oligodendrocytes), while SCs were characterized by immunostaining for S-100. Rats with transection injuries received scaffold implants containing NSCs (n=17), SCs (n=17), and no cells (control) (n=8). The degree of axonal regeneration was determined by counting neurofilament-stained axons through the scaffold channels 1 month after transplantation. Serial sectioning through the scaffold channels in NSC- and SC-treated groups revealed the presence of nestin, neurofilament, S-100, and betaIII tubulin-positive cells. GFAP-positive cells were only seen at the spinal cord-scaffold border. There were significantly more axons in the NSC- and SC- treated groups compared to the control group. In conclusion, biodegradable scaffolds with aligned columns seeded with NSCs or SCs facilitate regeneration across the transected spinal cord. Further, these multichannel biodegradable polymer scaffolds effectively serve as platforms for quantitative analysis of axonal regeneration. PMID- 19191515 TI - Absorption spectra of estradiol and tryptophan constructed by the statistical and elongation methods. AB - The statistical quantum chemical/molecular dynamical method is developed and employed to reproduce optical spectra. This technique includes quantum-mechanical calculations on energy states and photophysical properties of molecular conformers obtained during molecular dynamical simulation. Polycyclic organic molecule estradiol surrounded by solvent particles and protein structure including tryptophan fragment under thermodynamical conditions are considered. A wide absorption spectrum over several excited electronic states of estradiol is constructed. First longwave absorption band of tryptophan-cage mini protein is built involving the elongation method. These statistical spectra reflect the main features of the corresponding experimental ones. PMID- 19191516 TI - Electrostatic matching versus close-packing molecular arrangement in compressed dimethyl sulfoxide (DMSO) polymorphs. AB - Single crystals of dimethyl sulfoxide (DMSO, (CH3)2SO) were in situ frozen at isochoric conditions in a diamond-anvil cell and their structures determined at 0.37, 0.56, and 2.4 GPa. At ambient pressure, DMSO freezes at 291 K in monoclinic phase alpha, space group P2(1)/c, stable in all the temperature range from its melting point down to 2 K. On increasing the pressure, DMSO freezes at 140 MPa/296 K in phase alpha too, but above 540 MPa it collapses into a more compact triclinic phase beta, space group P1. The molecular aggregation in the crystal structure of DMSO is dominated by electrostatic attraction between negative and positive sites on the molecular surface and CH...O hydrogen bonds linking the molecules into dimers and chains. Most of this electrostatic matching and CH...O bonds present in phase alpha are preserved above 540 MPa, but the more tight packing in phase beta is achieved at the cost of broken dipole-dipole interactions between antiparallel SO groups and repulsing contacts between electropositive H-atoms squeezed to distances commensurate with the sum of van der Waals radii. The isostructural relation between phases alpha and beta is strictly connected with the molecular aggregation governed by electrostatic matching of interatomic contacts. PMID- 19191514 TI - Tensile stimulation of murine stem cell-collagen sponge constructs increases collagen type I gene expression and linear stiffness. AB - The objectives of this study were to determine how tensile stimulation delivered up to 14 days in culture influenced type I collagen gene expression in stem cells cultured in collagen sponges, and to establish if gene expression, measured using a fluorescence method, correlates with an established method, real-time quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). Using a novel model system, mesenchymal stem cells were harvested from six double transgenic mice in which the type I and type II collagen promoters were linked to green fluorescent protein-topaz and enhanced cyan fluorescent protein, respectively. Tissue-engineered constructs were created by seeding 0.5 x 10(6) mesenchymal stem cells onto type I collagen sponge scaffolds in a silicone dish. Constructs were then transferred to a custom pneumatic mechanical stimulation system housed in a standard incubator and stimulated for 5 h=day in tension for either 7 or 14 days using a repeated profile (2.4% peak strain for 20 s at 1 Hz followed by a rest period at 0% strain for 100 s). Control specimens were exposed to identical culture conditions but without mechanical stimulation. At three time points (0, 7, and 14 days), constructs were then prepared for evaluation of gene expression using fluorescence analysis and qRT-PCR, and the remaining constructs were failed in tension. Both analytical methods showed that constructs stimulated for 7 and 14 days showed significantly higher collagen type I gene expression than nonstimulated controls at the same time interval. Gene expression measured using qRT-PCR and fluorescence analysis was positively correlated (r = 0.9). Linear stiffness of stimulated constructs was significantly higher at both 7 and 14 days than that of nonstimulated controls at the same time intervals. Linear stiffness of the stimulated constructs at day 14 was significantly different from that of day 7. Future studies will vary the mechanical signal to optimize type I collagen gene expression to improve construct biomechanics and in vivo tendon repair. PMID- 19191517 TI - Regulating fibrinolysis to engineer skeletal muscle from the C2C12 cell line. AB - Muscles engineered from transformed cells would be a powerful model for the study of muscle physiology by allowing long-term in vitro studies of muscle adaptation. However, previously described methods either take >5 weeks to produce a tissue or use collagen as a scaffold, which decreases the specific force of the muscle, making it hard to measure the function of the constructs. The aim of this study was to rapidly engineer muscle using the C2C12 cell line in fibrin, which has a stiffness similar to muscle tissue, allowing accurate functional testing. Both the protease inhibitor aprotinin and the natural cross-linker genipin increased the length of time that muscle could be cultured, with genipin increasing the time in culture to 10 weeks. The function of the tissues was significantly affected by the batch of serum (64-78%) or thrombin (41%), the differentiation medium (78%), and the seeding protocol (38%), but was unaffected by initial cell number. Strikingly, different C2C12 clones produced up to a 3.6-fold variation in force production. Under optimal conditions, the tissues form in 10.4+/-0.3 days and remain fully functional for 5 weeks over which time they continue to mature. The optimized model described here provides rapid, reliable, and functional tissues that will be useful in the study of skeletal muscle physiology. PMID- 19191518 TI - Platelet lysate as a serum substitute for 2D static and 3D perfusion culture of stromal vascular fraction cells from human adipose tissue. AB - Fetal bovine serum (FBS) and fibroblast growth factor (FGF)-2 are key supplements for the culture of stromal vascular fraction (SVF) cells from adipose tissue, both for typical monolayer (2D) expansion and for streamlined generation of osteogenic-vasculogenic grafts in 3D perfusion culture. The present study investigates whether factors present in human platelet lysate (PL) could substitute for FBS and FGF-2 in 2D and 3D culture models of SVF cells from human lipoaspirates. SVF cells were grown in medium supplemented with 10% FBS+FGF-2 or with 5% PL. In 2D cultures, PL initially supported SVF cell proliferation, but resulted in growth arrest shortly after the first passage. Freshly isolated SVF cells cultured with both media under perfusion for 5 days within 3D ceramic scaffolds induced bone formation after subcutaneous implantation in nude mice. However, blood vessels of donor origin were generated only using FBS+FGF-2 cultured cells. This was unexpected, because the proportion of CD34+/CD31+ endothelial lineage cells was significantly higher with PL than that of FBS+FGF-2 (33% vs. 3%, respectively). These results support the use of PL as a substitute of FBS+FGF-2 for short-term culture of human SVF cells, and indicate that more specific serum-free formulations are required to maintain a functionally vasculogenic fraction of SVF cells expanded under 3D perfusion. PMID- 19191519 TI - Supramolecular modification of ion chemistry: modulation of peptide charge state and dissociation behavior through complexation with cucurbit[n]uril (n = 5, 6) or alpha-cyclodextrin. AB - Electrospray Fourier transform ion cyclotron resonance mass spectrometry, ion mobility spectrometry, and computational methods were utilized to characterize the complexes between lysine or pentalysine with three prototypical host molecules: alpha-cyclodextrin (alpha-CD), cucurbit[5]uril (CB[5]), and cucurbit[6]uril (CB[6]). Ion mobility measurements show lysine forms externally bound, singly charged complexes with either alpha-CD or CB[5], but a doubly charged complex with the lysine side chain threaded through the host cavity of CB[6]. These structural differences result in distinct dissociation behaviors in collision-induced dissociation (CID) experiments: the alpha-CD complex dissociates via the simple loss of intact lysine, whereas the CB[5] complex dissociates to yield [CB[5] + H(3)O](+), and the CB[6] complex loses neutral NH(3) and CO, the product ion remaining a doubly charged complex. These results are consistent with B3LYP/6-31G* binding energies (kJ mol(-1)) of D(Lys + H(+) alpha-CD) = 281, D(Lys + H(+)-CB[5]) = 327, and D(Lys + 2H(2+)-CB[6]) = 600. B3LYP/6-31G* geometry optimizations show complexation with alpha-CD stabilizes the salt bridge form of protonated lysine, whereas complexation with CB[6] stabilizes doubly protonated lysine. Complexation of the larger polypeptide pentalysine with alpha-CD forms a nonspecific adduct: no modification of the pentalysine charge state distribution is observed, and dissociation occurs via the simple loss of alpha-CD. Complexation of pentalysine with the cucurbiturils is more specific: the observed charge state distribution shifts higher on complexation, and fragmentation patterns are significantly altered relative to uncomplexed pentalysine: C-terminal fragment ions appear that are consistent with charge stabilization by the cucurbiturils, and the cucurbiturils are retained on the fragment ions. Molecular mechanics calculations suggest CB[5] binds to two protonated sites on pentalysine without threading onto the peptide and that CB[6] binds two adjacent protonated sites via threading onto the peptide. PMID- 19191520 TI - Solution structure of the DNA damage lesion 8-oxoguanosine from ultraviolet resonance Raman spectroscopy. AB - UV radiation and reactive byproducts of cellular metabolism are constant threats to genomic stability. A frequent consequence is the oxidation of DNA nucleobases, especially guanine to 8-oxoguanosine. This highly mutagenic lesion can form base pairs with other nucleobases, does not significantly distort the DNA structure, and remains unnoticed by DNA polymerases. Detection and biophysical studies of modified nucleobases is challenging because they are not fluorescent and have broad electronic spectra that overlap with those of normal bases. The structure of 8-oxoguanosine and its anion in solution has been contentious in the literature. Using ultraviolet excitation in resonance with the nucleobase, we have obtained the Raman spectra of 8-oxoguanosine. The stable tautomer in solution is unequivocally identified as the diketone form. We show that, at high pH, 8-oxoguanosine gets deprotonated to form an anion through loss of the N1 proton from the pyrimidine ring. The enol form is never populated to a detectable level. Raman spectra are supported by density functional theoretical calculations and a complete normal-mode analysis to identify bands that can be used as reporters of protein-nucleobase interactions. We have demonstrated that UVRR spectra provide unprecedented information on the solution-state structures of modified nucleobases. PMID- 19191521 TI - Excited states and photochemistry of bicyclo[1.1.0]butane. AB - Calculations on the excited states of bicyclo[1.1.0]butane in the gas phase by different theoretical methods using several basis sets were performed. In general, the agreement between calculated and experimental excitation energies for bicyclo[1.1.0]butane in the gas phase is very good. Reviews of the solution phase photochemistry of bicyclo[1.1.0]butane as well as previous calculations on the ground and excited states of bicyclo[1.1.0]butane are given to provide a necessary perspective of the photochemistry of bicyclo[1.1.0]butane in solution. To simulate the solution-phase photochemistry of bicyclo[1.1.0]butane, a well potential is added to the Kirkwood-Onsager model for obtaining solvation energies of molecules in solution. The addition of the well potential gives rise to a blue shift of all gas-phase excitation energies in solution. However, there is also the very important added effect of providing an increase in Rydberg-valence mixing of solution-phase excited states. It is this mixing of antibonding valence character into the solution-phase excited states that is necessary to explain the solution-phase photochemistry of bicyclo[1.1.0]butane through bond-breaking and the formation of a conical intersection intermediate. PMID- 19191522 TI - Theoretical study of mechanism of 2,3-dihydro-1,5-benzodiazepin-2-ones hydrazinolysis. AB - Density functional theory approach was used for the 4-methyl-2,3-dihydro-1,5 benzodiazepin-2-one compound to determine the mechanism of hydrazinolysis of 4 substituted 2,3-dihydro-1,5-benzodiazepin-2-ones. Single point computations at the MP2/6-311+G(d,p)//B3LYP/6-31G(d) level were performed for the more precise energy prediction. The solvent effect was taken into account by carrying out single point calculations using the PCM methodology. The obtained results show that in the investigating mechanism the first step consists of the hydrazine molecule addition to the azomethine bond of the 4-methyl-2,3-dihydro-1,5 benzodiazepin-2-one. Further cyclization occurs with pyrazole ring formation, and then the diazepine ring opening is revealed. Finally, removal of o phenylendiamine leads to 3-methylpyrazolone-5 as a main product that is in agreement with the experimental observation. The final step is a rate-determining step of this reaction. PMID- 19191523 TI - Characterization of scaffolds for tissue engineering by benchtop-magnetic resonance imaging. AB - Several publications have shown approaches for the optimization of tissue engineering constructs by magnetic resonance imaging (MRI). However, the technology is still scarcely used, probably because of the poor spatial resolution of clinical scanners and their temporally limited availability for many researchers. The new benchtop- MRI (BT-MRI) equipment used in the present study is much more affordable, for example, because of the low static magnetic field strength of 0.5 T and the absence of a helium cooling system. In this study, the method of BT-MRI was evaluated for the characterization of a tissue engineering scaffold. Hollow cylinder scaffolds were made of hydroxyapatite (HA), collagen, and chitosan and wrapped in a polyglycolic acid mesh. Mass transport between construct and surrounding medium was investigated by dynamic contrast agent-enhanced MRI with gadolinium(III)-diethylaminepentaacetic acid. The results demonstrate that BT-MRI permits detailed, space-resolved insights into diffusion processes within the three-dimensional matrices, enabling a comparison of the mass transport inside different scaffold types. Inhomogeneities of the HA distribution in scaffolds caused by the fabrication were also visible in MR images. The fate of cells, labeled with superparamagnetic iron oxide nanoparticles and seeded on the scaffold surface, was monitored. For the first time, it was shown that mass transport, inhomogeneities of the HA distribution, and localization of superparamagnetic iron oxide nanoparticle-labeled cells are accessible in a tissue engineering scaffold by BT-MRI. Hence, it is demonstrated that BT-MRI is a powerful analytic method for the noninvasive evaluation of tissue engineering constructs. PMID- 19191526 TI - Variants of MUC5B minisatellites and the susceptibility of bladder cancer. AB - The human MUC5B gene, which is primarily expressed in the tracheobronchial tract, is clustered to chromosome 11p15.5 with three other secreted gel-forming mucins, MUC6, MUC2, and MUC5AC. In this study, we identified seven variable number of tandem repeats (VNTRs; minisatellites) from the entire MUC5B region. Six (MUC5B MS1, -MS2, -MS3, -MS4, -MS5, and -MS7) of the seven minisatellites evaluated in this study were novel minisatellites, but the MUC5B-MS6 minisatellite was described in a previous study. These minisatellites of MUC5B were analyzed in genomic DNA extracted from controls, cancer patients, and multigenerational families. Three (MUC5B-MS3, -MS6, and -MS7) of the seven minisatellites were found to be polymorphic and transmitted through meiosis following Mendelian inheritance in seven families; therefore, these minisatellite polymorphisms could be useful as markers for paternity mapping and DNA fingerprinting. In addition, we evaluated allelic variation in these minisatellites to determine if such variation affected the susceptibility to various carcinomas. To accomplish this, we conducted a case-control study in which the genomic DNA of 789 cancer-free controls and cancer patients with five types of cancer were compared. A statistically significant association between the long rare MUC5B-MS6 alleles and the occurrence of bladder cancer was identified in the younger group (<60; odds ratio, 4.54; 95% confidence interval, 1.0-20.7; p=0.03). This observation suggests that the long rare MUC5B-MS6 alleles evaluated in this study could be used to identify the risk of bladder cancer. PMID- 19191524 TI - Design of highly stabilized beta-hairpin peptides through cation-pi interactions of lysine and n-methyllysine with an aromatic pocket. AB - Two tryptophan residues were incorporated on one face of a beta-hairpin peptide to form an aromatic pocket that interacts with a lysine or N-methylated lysine via cation-pi interactions. The two tryptophan residues were found to pack against the lysine side chain forming an aromatic pocket similar to those observed in trimethylated lysine receptor proteins. Thermal analysis of methylated lysine variant hairpin peptides revealed an increase in thermal stability as the degree of methylation was increased, resulting in the most thermally stable beta-hairpin reported to date. PMID- 19191527 TI - Computational modeling study on formation of acyclic clavulanate intermediates in inhibition of class A beta-lactamase: water-assisted proton transfer. AB - Molecular dynamics (MD) simulation and quantum chemical (QC) calculations were used to investigate the reaction mechanism of the formation of acyclic clavulanate intermediates in the inhibition of class A beta-lactamase. The initial model for QC calculations was derived from an MD simulation. It was composed of a substrate clavulanate and four residues (Ser70, Gln237, Ser130, and Ser216), which form hydrogen bonds with the substrate. The QC calculation results indicate that the oxazolidine ring can undergo cleavage by proton transfer, which yields not only imine but also enamine products. A new mechanism involving hydrogen transfer from C6 to O1 has been suggested. Besides, MD simulation provided evidence that the water molecule can catalyze the proton transfer, and QC calculation shows water assistance can decrease the energy barrier greatly. PMID- 19191536 TI - Astrobiological polarimeter. AB - Chirality is an excellent indicator of life, but naturally occurring astrobiological (as well as terrestrial) samples nearly always exhibit massive depolarizing light scattering, which renders conventional polarimeters useless. For astrobiological applications, we instead consider a novel polarimeter originally developed for non-invasive human-glucose measurement. It involves deliberately rotating in time the plane of polarization of a linearly polarized beam and detecting the shift in the plane of the rotating linearly polarized component of the transmitted light from a chiral sample relative to the input polarization plane. We find that this polarimeter can operate in 3 orders of magnitude more depolarizing scattering than conventional polarimeters. Furthermore, it can also be designed to be lightweight, compact, and energy efficient. PMID- 19191537 TI - Novel Sfp1 transcriptional regulation of Saccharomyces cerevisiae gene expression changes during spaceflight. AB - This study identifies transcriptional regulation of stress response element (STRE) genes in space in the model eukaryotic organism, Saccharomyces cerevisiae. To determine transcription-factor dependence, gene expression changes in space were examined in strains bearing green fluorescent protein-tagged (GFP-tagged) reporters for YIL052C (Sfp1 dependent with stress), YST-2 (Sfp1/Rap1 dependent with stress), or SSA4 (Msn4 dependent with stress), along with strains of SSA4 GFP and YIL052C-GFP with individual deletions of the Msn4 or Sfp1. When compared to parallel ground controls, spaceflight induces significant gene expression changes in SSA4 (35% decrease) and YIL052C (45% decrease), while expression of YST-2 (0.08% decrease) did not change. In space, deletion of Sfp1 reversed the SSA4 gene expression effect (0.00% change), but Msn4 deletion yielded a similar decrease in SSA4 expression (34% change), which indicates that SSA4 gene expression is dependent on the Sfp1 transcription factor in space, unlike other stresses. For YIL052C, deletion of Sfp1 reversed the effect (0.01% change), and the Msn4 deletion maintained the decrease in expression (30% change), which indicates that expression of YIL052C is also dependent on Sfp1 in space. Spaceflight has selective and specific effects on SSA4 and YIL052C gene expression, indicated by novel dependence on Sfp1. PMID- 19191538 TI - Short- and long-term olivine weathering in Svalbard: implications for Mars. AB - Liquid water is essential to life as we know it on Earth; therefore, the search for water on Mars is a critical component of the search for life. Olivine, a mineral identified as present on Mars, has been proposed as an indicator of the duration and characteristics of water because it dissolves quickly, particularly under low-pH conditions. The duration of olivine persistence relative to glass under conditions of aqueous alteration reflects the pH and temperature of the reacting fluids. In this paper, we investigate the utility of 3 methodologies to detect silicate weathering in a Mars analog environment (Sverrefjell volcano, Svalbard). CheMin, a miniature X-ray diffraction instrument developed for flight on NASA's upcoming Mars Science Laboratory, was deployed on Svalbard and was successful in detecting olivine and weathering products. The persistence of olivine and glass in Svalbard rocks was also investigated via laboratory observations of weathered hand samples as well as an in situ burial experiment. Observations of hand samples are consistent with the inference that olivine persists longer than glass at near-zero temperatures in the presence of solutions at pH approximately 7-9 on Svalbard, whereas in hydrothermally altered zones, glass has persisted longer than olivine in the presence of fluids at similar pH at approximately 50 degrees C. Analysis of the surfaces of olivine and glass samples, which were buried on Sverrefjell for 1 year and then retrieved, documented only minor incipient weathering, though these results suggest the importance of biological impacts. The 3 types of observations (CheMin, laboratory observations of hand samples, burial experiments) of weathering of olivine and glass at Svalbard show promise for interpretation of weathering on Mars. Furthermore, the weathering relationships observed on Svalbard are consistent with laboratory-measured dissolution rates, which suggests that relative mineral dissolution rates in the laboratory, in concert with field observations, can be used to yield valuable information regarding the pH and temperature of reacting martian fluids. PMID- 19191539 TI - Inorganic nitrogen reduction and stability under simulated hydrothermal conditions. AB - Availability of reduced nitrogen is considered a prerequisite for the genesis of life from prebiotic precursors. Most atmospheric and oceanic models for the Hadean Earth predict a mildly oxidizing environment that is conducive to the formation and stability of only oxidized forms of nitrogen. A possible environment where reduction of oxidized nitrogen to ammonium has been speculated to occur is aqueous hydrothermal systems. We examined a suite of transition metal oxides and sulfides for their ability to reduce nitrate and nitrite, as well as oxidize ammonia, under hot (300 degrees C) high-pressure (50-500 MPa) aqueous conditions. In general, iron sulfides exhibited the most rapid and complete conversion noted, followed by nickel and copper sulfides to a much lower degree. Of the oxides examined, only magnetite exhibited any ability to reduce NO(3)(-) or NO(2)(-). Ammonium was stable or exhibited small losses (<20%) in contact with all the mineral phases and conditions tested. The results support the idea that hydrothermal systems could have provided significant amounts of reduced nitrogen to their immediate environments. The enhanced availability of reduced nitrogen in hydrothermal systems also has important implications for prebiotic metabolic pathways where nitrogen availability is critical to the production of amino acids and other nitrogenous compounds. PMID- 19191540 TI - Fossilized microorganisms from the Emperor Seamounts: implications for the search for a subsurface fossil record on Earth and Mars. AB - We have observed filamentous carbon-rich structures in samples drilled at 3 different seamounts that belong to the Emperor Seamounts in the Pacific Ocean: Detroit (81 Ma), Nintoku (56 Ma), and Koko Seamounts (48 Ma). The samples consist of low-temperature altered basalts recovered from all 3 seamounts. The maximum depth from which the samples were retrieved was 954 meters below seafloor (mbsf). The filamentous structures occur in veins and fractures in the basalts, where they are attached to the vein walls and embedded in vein-filling minerals like calcite, aragonite, and gypsum. The filaments were studied with a combination of optical microscopy, environmental scanning electron microscopy (ESEM), Raman spectroscopy, and time-of-flight secondary ion mass spectrometry (ToF-SIMS). Minerals were identified by a combination of optical microscopy, X-ray diffraction, Raman spectrometry, and energy dispersive spectrometry on an environmental scanning electron microscope. Carbon content of the filaments ranges between approximately 10 wt % and approximately 50 wt % and is not associated with carbonates. These results indicate an organic origin of the carbon. The presence of C(2)H(4), phosphate, and lipid-like molecules in the filaments further supports a biogenic origin. We also found microchannels in volcanic glass enriched in carbon (approximately 10-40 wt %) compatible with putative microbial activity. Our findings suggest new niches for life in subseafloor environments and have implications for further exploration of the subseafloor biosphere on Earth and beyond. PMID- 19191541 TI - Isolation of rpoB mutations causing rifampicin resistance in Bacillus subtilis spores exposed to simulated Martian surface conditions. AB - ABSTRACT Bacterial spores are considered prime candidates for Earth-to-Mars transport by natural processes and human spaceflight activities. Previous studies have shown that exposure of Bacillus subtilis spores to ultrahigh vacuum (UHV) characteristic of space both increased the spontaneous mutation rate and altered the spectrum of mutation in various marker genes; but, to date, mutagenesis studies have not been performed on spores exposed to milder low pressures encountered in the martian environment. Mutations to rifampicin-resistance (Rif(R)) were isolated in B. subtilis spores exposed to simulated martian atmosphere (99.9% CO(2), 710 Pa) for 21 days in a Mars Simulation Chamber (MSC) and compared to parallel Earth controls. Exposure in the MSC reduced spore viability by approximately 67% compared to Earth controls, but this decrease was not statistically significant (P = 0.3321). The frequency of mutation to Rif(R) was also not significantly increased in the MSC compared to Earth-exposed spores (P = 0.479). Forty-two and 51 Rif(R) mutant spores were isolated from the MSC- and Earth-exposed controls, respectively. Nucleotide sequencing located the Rif(R) mutations in the rpoB gene encoding the beta subunit of RNA polymerase at residue V135F of the N-cluster and at residues Q469K/L, H482D/P/R/Y, and S487L in Cluster I. No mutations were found in rpoB Clusters II or III. Two new alleles, Q469L and H482D, previously unreported in B. subtilis rpoB, were isolated from spores exposed in the MSC; otherwise, only slight differences were observed in the spectra of spontaneous Rif(R) mutations from spores exposed to Earth vs. the MSC. However, both spectra are distinctly different from Rif(R) mutations previously reported arising from B. subtilis spores exposed to simulated space vacuum. PMID- 19191542 TI - Determination of lethality rate constants and D-values for Bacillus atrophaeus (ATCC 9372) spores exposed to dry heat from 115 degrees C to 170 degrees C. AB - Dry heat microbial reduction is the NASA-approved sterilization method to reduce the microbial bioburden on spaceflight hardware for missions with planetary protection requirements. The method involves heating the spaceflight hardware to temperatures between 104 degrees C and 125 degrees C for up to 50 hours, while controlling the humidity to very low values. Collection of lethality data at temperatures above 125 degrees C and with ambient (uncontrolled) humidity conditions would establish whether any microbial reduction credit can be offered to the flight project for processes that occur at temperatures greater than 125 degrees C. The goal of this research is to determine the survival rates of Bacillus atrophaeus (ATCC 9372) spores subjected to temperatures higher than 125 degrees C under both dry (controlled) and room ambient humidity (36-66% relative humidity) conditions. Spores were deposited inside thin, stainless steel thermal spore exposure vessels (TSEVs) and heated under ambient or controlled humidity conditions from 115 degrees C to 170 degrees C. After the exposures, the TSEVs were cooled rapidly, and the spores were recovered and plated. Survivor ratios, lethality rate constants, and D-values were calculated at each temperature. At 115 degrees C and 125 degrees C, the controlled humidity lethality rate constant was faster than the ambient humidity lethality rate constant. At 135 degrees C, the ambient and controlled humidity lethality rate constants were statistically identical. At 150 degrees C and 170 degrees C, the ambient humidity lethality rate constant was slightly faster than the controlled humidity lethality rate constant. These results provide evidence for possibly modifying the NASA dry heat microbial reduction specification. PMID- 19191544 TI - Changes of cervical dorsal root ganglia induced by compression injury and decompression procedure: a novel rat model of cervical radiculoneuropathy. AB - Our study aimed to establish a model of compression injury of cervical dorsal root ganglia (DRG) in the rat and to investigate the pathological changes following compression injury and decompression procedures. Thirty rats were divided into three groups: control group receiving sham surgery, compression group undergoing surgery to place a micro-silica gel on C6 DRG, and decompression group with subsequent decompression procedure. The samples harvested from the different groups were examined with light microscopy, ultrastructural analysis, and horseradish peroxidase (HRP) retrograde tracing techniques. Apoptosis of DRG neurons was demonstrated with TUNEL staining. Changes in PGE2 and PLA2 in DRG neurons were detected with enzyme-linked immunosorbent assay (ELISA). Local expression of vascular endothelial growth factor (VEGF) was monitored with immunohistochemistry. DRG neurons in the compression group became swollen with vacuolar changes in cytoplasm. Decompression procedure partially ameliorated the resultant compression pathology. Ultrastructural examination showed a large number of swollen vacuoles, demyelinated nerve root fibers, absence of Schwann cells, and proliferation in the surrounding connective tissues in the compression group. Compared to the control group, the compression group showed a significant decrease in the number of the HRP-labeled cells and a significant increase in levels of PGE2 and PLA2, in the expression of VEGF protein, and in the number of apoptotic DRG neurons. These findings demonstrate that compression results in local inflammation, followed by increased apoptosis and upregulation of VEGF. We conclude that such a model provides a tool to study the pathogenesis and treatment of cervical radiculoneuropathy. PMID- 19191546 TI - Loop-tryptophan human purine nucleoside phosphorylase reveals submillisecond protein dynamics. AB - Human PNP is a homotrimer containing three tryptophan residues at positions 16, 94, and 178, all remote from the catalytic site. The catalytic sites of PNP are located near the subunit-subunit interfaces where F159 is a catalytic site residue donated from an adjacent subunit. F159 covers the top (beta) surface of the ribosyl group at the catalytic site. QM/MM calculations of human PNP have shown that F159 is the center of the most mobile region of the protein providing access to the substrate in the active site. F159 is also the key residue in a cluster of hydrophobic residues that shield catalytic site ligands from bulk solvent. Trp-free human PNP (Leuko-PNP) was previously engineered by replacing the three Trp residues of native PNP with Tyr. From this active construct, a single Trp residue was placed in the catalytic site loop (F159W-Leuko-PNP) as a reporter group for the ribosyl region of the catalytic site. The F159W-Leuko-PNP fluorescence is red shifted compared to native PNP, suggesting a solvent-exposed Trp residue. Upon ligand binding (hypoxanthine), the 3-fold fluorescence quench confirms conformational packing of the catalytic site pocket hydrophobic cluster. F159W-Leuko-PNP has an on-enzyme thermodynamic equilibrium constant (Keq) near unity in the temperature range between 20 and 30 degrees C and nonzero enthalpic components, making it suitable for laser-induced T-jump analyses. T-jump relaxation kinetics of F159W-Leuko-PNP in equilibrium with substrates and/or products indicate the conformational equilibria of at least two ternary complex intermediates in the nano- to millisecond time scale (1000-10000 s-1) that equilibrate prior to the slower chemical step (approximately 200 s-1). F159W Leuko-PNP provides a novel protein platform to investigate the protein conformational dynamics occurring prior to transition state formation. PMID- 19191548 TI - Delineation of alternative conformational states in Escherichia coli peptide deformylase via thermodynamic studies for the binding of actinonin. AB - We investigated the binding of a naturally occurring antibiotic, actinonin, to the Ni(2+)-reconstituted recombinant form of Escherichia coli peptide deformylase (PDF(Ec)) via isothermal titration microcalorimetry. The binding data conformed to both exothermic and endothermic phases with magnitudes of DeltaG degrees , DeltaH degrees , and TDeltaS degrees being equal to -12, -2.7, and 9.3 kcal/mol and -8.7, 3.9, and 12.6 kcal/mol, respectively. Evidently, although both phases are dominated by favorable entropic changes, the exothermic phase has about 6.7 kcal/mol enthalpic advantage over the endothermic phase. We observed that the removal of bound Ni(2+) from PDF(Ec) abolished the exothermic phase without affecting the endothermic phase, but it was regained upon addition of Zn(2+). In conjunction with metal analysis data, we propose that the recombinant form of PDF(Ec) is expressed in two stable conformational states that yield markedly distinct ITC profiles (i.e., exothermic versus endothermic) upon interaction with actinonin. The existence of two conformational states of PDF(Ec) is further supported by the observation of two distinct and independent transitions during the thermal unfolding of the enzyme. In addition, the thermodynamic data reveal that the formation of the PDF(Ec)-actinonin complex results in the transfer of one H(+) from the enzyme phase to the bulk solvent at pH 6.3. Both exothermic and endothermic phases produce highly negative DeltaC(p) degrees values, but there is no apparent enthalpy-entropy compensation effect upon formation of the PDF(Ec) actinonin complex. In view of the known structural features of the enzyme, arguments are presented that the alternative conformational states of PDF(Ec) are modulated by the metal ligation at the enzyme site. PMID- 19191549 TI - Neopyrrolomycins with broad spectrum antibacterial activity. AB - Three new antibiotics, neopyrrolomycins B (1), C (2), and D (3), with potent activity against Gram-positive pathogens were discovered. They exhibited MIC values < 1 microg/mL versus a number of resistant strains. The compounds were obtained from the ethyl acetate extracts of a Streptomyces sp. after purification by column chromatography and RP-HPLC. Their structures were elucidated using X ray crystallography (1) and NMR spectroscopy (2 and 3). PMID- 19191550 TI - Rapid cloning and heterologous expression of the meridamycin biosynthetic gene cluster using a versatile Escherichia coli-streptomyces artificial chromosome vector, pSBAC. AB - Expression of biosynthetic pathways in heterologous hosts is an emerging approach to expedite production improvement and biosynthetic modification of natural products derived from microbial secondary metabolites. Herein we describe the development of a versatile Escherichia coli-Streptomyces shuttle Bacterial Artificial Chromosomal (BAC) conjugation vector, pSBAC, to facilitate the cloning, genetic manipulation, and heterologous expression of actinomycetes secondary metabolite biosynthetic gene clusters. The utility of pSBAC was demonstrated through the rapid cloning and heterologous expression of one of the largest polyketide synthase (PKS) and nonribosomal peptide synthetase (NRPS) biosynthetic pathways: the meridamycin biosynthesis gene cluster (mer). The entire mer gene cluster ( approximately 90 kb) was captured in a single pSBAC clone through a straightforward restriction enzyme digestion and cloning approach and transferred into Streptomyces lividans. The production of meridamycin (1) in the heterologous host was achieved after replacement of the original promoter with an ermE* promoter and was enhanced by feeding with a biosynthetic precursor. The success of heterologous expression of such a giant gene cluster demonstrates the versatility of BAC cloning technology and paves the road for future exploration of expression of the meridamycin biosynthetic pathway in various hosts, including strains that have been optimized for polyketide production. PMID- 19191551 TI - Alstiphyllanines A-D, indole alkaloids from Alstonia macrophylla. AB - Four new alkaloids, alstiphyllanines A-D (1-4), were isolated from Alstonia macrophylla, and their structures were determined by MS and 2D NMR analyses. Alkaloids 1-4 showed moderate antiplasmodial activity against Plasmodium falciparum and vasorelaxant activity against phenylephrine-induced contraction of isolated rat aorta. PMID- 19191554 TI - Discovery of N-(3-fluorophenyl)-1-[(4-([(3S)-3-methyl-1 piperazinyl]methyl)phenyl)acetyl]-4-piperidinamine (GSK962040), the first small molecule motilin receptor agonist clinical candidate. AB - N-(3-fluorophenyl)-1-[(4-([(3S)-3-methyl-1-piperazinyl]methyl)phenyl)acetyl]-4 piperidinamine 12 (GSK962040) is a novel small molecule motilin receptor agonist. It possesses excellent activity at the recombinant human motilin receptor and also at the native rabbit motilin receptor where its agonist activity results in potentiation of the amplitude of neuronal-mediated contractions of isolated gastric antrum tissue. Compound 12 also possesses highly promising pharmacokinetic profiles in both rat and dog, and these results, in combination with further profiling in human native tissue and an in vivo model of gastrointestinal transit in the rabbit, have led to its selection as a candidate for further development. PMID- 19191553 TI - Toward optimization of the second aryl substructure common to transthyretin amyloidogenesis inhibitors using biochemical and structural studies. AB - Transthyretin (TTR) amyloidogenesis inhibitors are typically composed of two aromatic rings and a linker. We have previously established optimal structures for one aromatic ring and the linker. Herein, we employ a suboptimal linker and an optimal aryl-X substructure to rank order the desirability of aryl-Z substructures--using a library of 56 N-(3,5-dibromo-4-hydroxyphenyl)benzamides. Coconsideration of amyloid inhibition potency and ex vivo plasma TTR binding selectivity data reveal that 2,6, 2,5, 2, 3,4,5, and 3,5 substituted aryls bearing small substituents generate the most potent and selective inhibitors, in descending order. These benzamides generally lack undesirable thyroid hormone receptor binding and COX-1 inhibition activity. Three high-resolution TTR.inhibitor crystal structures (1.31-1.35 A) provide insight into why these inhibitors are potent and selective, enabling future structure-based design of TTR kinetic stabilizers. PMID- 19191556 TI - Stereoselective synthesis of chiral IBR2 analogues. AB - Two stereoselective routes were developed to synthesize optically pure IBR2 analogues 1-16. The first features addition of N-Boc-3-bromoindole 26 to the sulfinamide 25, providing a 1:1 ratio of the separable diasteroisomers 27 and 28 in good yield. In a straightforward fashion, the sulfinamides 27 and 28 were conveniently converted into the key amines 39 and 47 over 8 steps, respectively, from which a series of 3,4-dihydroisoquinolinyl IBR2 analogues 1-14 containing fluorinated and trifluoromethylated benzyl groups were prepared. Another route highlights the highly enantioselective addition of indole to the sulfonyl amide 50 with bifunctional aminothioureas 57 and 58 as catalysts. After the reaction conditions were optimized, the desired sulfonyl amides (R)-55 and (S)-55 were obtained in 99% ee and 98% ee, respectively. Acylation of (R)-55 and (S)-55 separately and subsequent allylation gave compounds 60 and 63, respectively, which were further subjected to RCM to furnish compounds 61 and 64 and, after removal of the Boc groups, the desired IBR2 analogues 15 and 16. PMID- 19191557 TI - Phosphonosulfonates are potent, selective inhibitors of dehydrosqualene synthase and staphyloxanthin biosynthesis in Staphylococcus aureus. AB - Staphylococcus aureus produces a golden carotenoid virulence factor called staphyloxanthin (STX), and we report here the inhibition of the enzyme, dehydrosqualene synthase (CrtM), responsible for the first committed step in STX biosynthesis. The most active compounds are halogen-substituted phosphonosulfonates, with K(i) values as low as 5 nM against the enzyme and IC(50) values for STX inhibition in S. aureus as low as 11 nM. There is, however, only a poor correlation (R(2) = 0.27) between enzyme and cell pIC(50) (= -log(10) IC(50)) values. The ability to predict cell from enzyme data improves considerably (to R(2) = 0.72) with addition of two more descriptors. We also investigated the activity of these compounds against human squalene synthase (SQS), as a counterscreen, finding several potent STX biosynthesis inhibitors with essentially no squalene synthase activity. These results open up the way to developing potent and selective inhibitors of an important virulence factor in S. aureus, a major human pathogen. PMID- 19191558 TI - Cell-penetrating metal complex optical probes: targeted and responsive systems based on lanthanide luminescence. AB - To understand better the structure and function of biological systems, cell biologists and biochemists would like to have methods that minimally perturb living systems. The development of emissive optical probes is essential for improving our observation of intracellular signaling and recognition processes. Following excitation of the probe, photons emitted from the probe may be observed by spectroscopy or microscopy and encode information about their environments in their energy, lifetime, and polarization. Such optical probes may be based on organic fluorophores, quantum dots, recombinant proteins, or emissive metal complexes. In this Account, we trace the emergence of lanthanide coordination complexes as emissive optical probes. These probes benefit from sharp emission bands and long lifetimes. We can design these complexes to report on the concentration of key biochemical variables by modulation of spectral form, lifetime, or circular polarization. These properties allow us to apply ratiometric methods of analysis in spectroscopy or microscopy to report on local pH, pM (M = Ca, Zn), or the concentration of certain anionic metabolites, such as citrate, lactate, bicarbonate, or urate. For optical microscopy studies in living cells, these probes must be cell-permeable and, ideally, should localize in a given cell organelle. We undertook systematic studies of more than 60 emissive complexes, examining the time dependence of cellular uptake and compartmentalization, cellular toxicity, protein affinity, and quenching sensitivity. These results and their relationship to probe structure have allowed us to identify certain structure-activity relationships. The nature and linkage mode of the integral sensitizing group-introduced to harvest incident light efficiently-is of primary importance in determining protein affinity and cellular uptake and trafficking. In many cases, uptake may occur via macropinocytosis. We have defined three main classes of behavior: complexes exhibit predominant localization profiles in protein-rich regions (nucleoli/ribosomes), in cellular mitochondria, or in endosomes/lysosomes. Therefore, these systems offer considerable promise as intracellular optical probes, amenable to single- or two photon excitation, that may report on the local ionic composition of living cells subjected to differing environmental stresses. PMID- 19191559 TI - Fatty acid profile of bovine milk naturally enhanced with docosahexaenoic acid. AB - Recent studies have shown that the fatty acid profile of dietary lipid has the potential for improving the health of consumers. The present study was conducted to determine the fatty acid composition of commercial milks, namely, Dairy-Oh! Homo-Milk (DOHM), which is naturally enhanced with docosahexaenoic acid (DHA), or regular Homo-Milk (HM). The milk was collected from local supermarkets. The most abundant saturated fatty acids in the milk were butyric (C4:0), lauric (C12:0), myristic (C14:0), palmitic (C16:0), and stearic (C18:0) acids. Among unsaturated fatty acids, oleic acid (cis-9-C18:1) was also considerably high (502.7 mg/100 mL of milk). The concentration of total trans-18:1 was higher (P < 0.05) in DOHM than in HM (134.7 vs 107.0 mg/100 mL of milk, respectively), whereas total cis 18:1 was higher (P < 0.05) in HM than in DOHM (566.4 vs 508.4 mg/100 mL of milk, respectively). The concentration of DHA was 24.0 times higher (P < 0.05) in DOHM than in HM. DOHM contained 2.8 times higher (P < 0.05) eicosapentaenoic acid (EPA) compared to HM. Milk fat from DOHM contained a greater concentration of cis 9,trans-11 conjugated linoleic acid (CLA, 16.4 vs 11.6 mg/100 mL of milk, DOHM vs HM, respectively). The total omega-3 polyunsaturated fatty acids content was 2.23 times greater (P < 0.05) in DOHM compared with HM, due to an increase in C18:3n 3, EPA, and DHA. The result of the milk fatty acid analyses indicates that milk fat from DOHM had increased contents of EPA, DHA, and cis-9,trans-11 CLA, which could have a more favorable impact on diet composition and healthfulness. PMID- 19191561 TI - Catechols as membrane anion transporters. AB - We report that an amphiphilic bis-catechol (3) functions as a transmembrane anion transporter. Activity depends on the catechol's substitution and amphiphilicity. We also describe a liposomal assay that allows one to readily measure anion transport selectivity. This assay reveals that anion transport selectivity for this amphiphilic bis-catechol follows a Hofmeister sequence wherein anions that are easier to dehydrate are made more permeable to the membrane by 3. PMID- 19191560 TI - Characterization of the tautomycetin biosynthetic gene cluster from Streptomyces griseochromogenes provides new insight into dialkylmaleic anhydride biosynthesis. AB - Tautomycetin (TTN) is a highly potent and specific protein phosphatase inhibitor isolated from Streptomyces griseochromogenes. The biological activity of TTN makes it an important lead for drug discovery, whereas its rare dialkylmaleic anhydride moiety and structural similarity to tautomycin (TTM), another potent phosphatase inhibitor with tremendous medicinal potential, draws attention to novel biosynthetic chemistries responsible for its production. To elucidate the biosynthetic machinery associated with TTN production, the ttn biosynthetic gene cluster from S. griseochromogenes was isolated and characterized, and its involvement in TTN biosynthesis confirmed by gene inactivation and complementation experiments. The ttn cluster was localized to a 79 kb DNA region, consisting of 19 open reading frames that encode two modular type I polyketide synthases (TtnAB), one type II thioesterase (TtnH), eight proteins for dialkylmaleic anhydride biosynthesis (TtnKLMNOPRS), four tailoring enzymes (TtnCDFI), two regulatory proteins (TtnGQ), and one resistance protein (TtnJ). A model for TTN biosynthesis is proposed on the basis of functional assignments from sequence analysis, which agrees well with previous feeding experiments, has been supported by in vivo gene inactivation experiments, and is supported by analogy to the recently reported ttm cluster. These findings set the stage to fully investigate TTN biosynthesis and to biosynthetically engineer new TTN analogues. PMID- 19191562 TI - Antitumor psoropermum xanthones and sarcomelicope acridones: privileged structures implied in DNA alkylation. AB - Fused isopropylfuran and dimethylpyran units are privileged structures present in numerous bioactive natural products exemplified, in the field of anticancer drugs, by the furanoxanthone psorospermin and the pyranoacridone acronycine. Psorospermin binds to the N-7 position of the guanine units in the presence of topoisomerase II. In contrast, acronycine derivatives such as cis-1,2-diacetoxy 1,2-dihydrobenzo[b]acronycine alkylate the 2-amino group of DNA guanine residues in the minor groove. Hybrid compounds associating the acridone or benzo[b]acridone chromophore of acronycine derivatives and the epoxyfuran alkylating unit present in psorospermin also display very potent antiproliferative activities, alkylating DNA guanine units at position N-7 in the major groove, as natural xanthones belonging to the psorospermin series. PMID- 19191563 TI - Motualevic acids A-F, antimicrobial acids from the sponge Siliquariaspongia sp. AB - Seven new antibacterials, motualevic acids A-F (1-6) and (4E)-(R)-antazirine (7), have been isolated from the marine sponge Siliquariaspongia sp. and their structures elucidated by spectroscopic methods. Motualevic acids A-D are the first glycyl conjugates of the omega-brominated lipid (E)-14,14-dibromotetradeca 2,13-dienoic acid, and motualevic acid F is the first long-chain 2H-azirine 2 carboxylic acid to be found in nature. Carboxylic acid-containing compounds 1 and 6 inhibit the growth of Staphylococcus aureus and methicillin-resistant S. aureus at 1.2-10.9 microg/mL. PMID- 19191564 TI - Design of core--shell-type nanoparticles carrying stable radicals in the core. AB - Utilizing the self-assembled core-shell-type polymeric micelle technique, high performance nanoparticles possessing stable radicals in the core and reactive groups on the periphery were prepared. The anionic ring-opening polymerization of ethylene oxide (EO) was carried out using potassium 3,3-diethoxypropanolate as an initiator, followed by mesylation with methanesulfonyl chloride to obtain acetal poly(ethylene glycol)-methanesulfonate (acetal-PEG-Ms; 1). Compound 1 was reacted with potassium O-ethyldithiocarbonate, followed by treatment with n-propylamine to obtain heterobifunctional PEG derivatives containing both sulfanyl and acetal terminal groups (acetal-PEG-SH) (2) in a highly selective and quantitative manner. Poly(ethylene glycol)-block-poly(chloromethylstyrene) (acetal-PEG-b-PCMS) (3) was synthesized by the free-radical telomerization of chloromethylstyrene (CMS) using 2 as a telogen. The chloromethyl groups in the PCMS segment of the block copolymer (3) were quantitatively converted to 2,2,6,6 tetramethylpiperidinyloxys (TEMPOs) via the amination of 3 with 4-amino-TEMPO to obtain acetal-PEG-b-PCMS containing TEMPO moieties (4). The obtained 4 formed core-shell-type nanoparticles in aqueous media when subjected to the dialysis method: the cumulant average diameter of the nanoparticles was about 40 nm, and the nanoparticles emitted intense electron paramagnetic resonance (EPR) signals. The TEMPO radicals in the core of the nanoparticles showed reduction resistance even in the presence of 3.5 mM ascorbic acid. This means that these nanoparticles are anticipated as high-performance bionanoparticles that can be used in vivo. PMID- 19191565 TI - Compared to purpurinimides, the pyropheophorbide containing an iodobenzyl group showed enhanced PDT efficacy and tumor imaging (124I-PET) ability. AB - Two positional isomers of purpurinimide, 3-[1'-(3-iodobenzyloxyethyl)] purpurin 18-N-hexylimide methyl ester 4, in which the iodobenzyl group is present at the top half of the molecule (position-3), and a 3-(1'-hexyloxyethy)purpurin-18-N-(3 iodo-benzylimide)] methyl ester 5, where the iodobenzyl group is introduced at the bottom half (N-substitued cyclicimide) of the molecule, were derived from chlorophyll-a. The tumor uptake and phototherapeutic abilities of these isomers were compared with the pyropheophorbide analogue 1 (lead compound). These compounds were then converted into the corresponding 124I-labeled PET imaging agents with specific activity >1 Ci/micromol. Among the positional isomers 4 and 5, purpurinimide 5 showed enhanced imaging and therapeutic potential. However, the lead compound 1 derived from pyropheophorbide-a exhibited the best PET imaging and PDT efficacy. For investigating the overall lipophilicity of the molecule, the 3-O-hexyl ether group present at position-3 of purpurinimide 5 was replaced with a methyl ether substituent, and the resulting product 10 showed improved tumor uptake, but due to its significantly higher uptake in the liver, spleen, and other organs, a poor tumor contrast in whole-body tumor imaging was observed. PMID- 19191567 TI - Nonnatural amino acids for site-specific protein conjugation. AB - Over the years, several chemical reactions have been developed that enable the covalent conjugation of synthetic molecules to natural proteins. The resulting bioconjugates have become important tools in the study of natural proteins. Furthermore, they form a new class of protein-based pharmaceuticals and biomaterials. However, classical bioconjugation reactions to natural amino acids suffer from poor site-specificity. To overcome this problem, a variety of uniquely reactive non-natural amino acids have recently been designed. These can be incorporated into proteins by specifically engineered bacterial strains. Such reactive non-natural amino acids create new possibilities for bio-orthogonal conjugation to proteins. This review first gives an overview of the various methods for site-specific introduction of non-natural amino acids into proteins. Both semisynthetic and entirely recombinant methods are addressed. Then, a detailed description is given of the reactive non-natural amino acids that have already been recombinantly introduced into proteins. The bio-orthogonal reactions that can be used for conjugation to these reactive non-natural amino acids are also discussed. These include the alkyne/azide 'click' reaction, carbonyl condensations, Michael-type additions, and Mizoroki-Heck substitutions. PMID- 19191566 TI - Design and synthesis of biomimetic hydrogel scaffolds with controlled organization of cyclic RGD peptides. AB - We report on the rational design and synthesis of a new type of bioactive poly(ethylene glycol) diacrylate (PEGDA) macromers, cyclic Arg-Gly-Asp (cRGD) PEGDA, to mimic the cell-adhesive properties of extracellular matrix (ECM), aiming to create biomimetic scaffolds with controlled spatial organization of ligands and enhanced cell binding affinity for tissue engineering. To attach the cRGD peptide in the middle of PEGDA chain, a tailed cRGD peptide, c[RGDfE(SSSKK NH2)] (1), was synthesized with c(RGDfE) linked to a tail of SSSKK. The tail consists of a spacer with three serine residues and a linker with two lysine residues for conjugating with acryloyl-PEG-NHS (5) to create cRGD-PEGDA (6). cRGD PEGDA possesses good photopolymerization ability to fabricate hydrogel scaffolds under UV radiation. Surface morphology and composition analysis demonstrates that cRGD-PEGDA hydrogels were well-constructed with porous three-dimensional (3D) structures and uniform distribution of cRGD ligands. Our results show that cRGD PEGDA hydrogels facilitate endothelial cell (EC) adhesion and spreading on the hydrogel surfaces and exhibit significantly higher EC population in comparison with linear RGD-modified hydrogels at low peptide incorporation. Since ligand presentation in biomimetic scaffolds plays an important role in controlling cell behavior, cRGD-PEGDA has great advantages of controlling hydrogel properties and ligand spatial organization in the resulting scaffolds. Furthermore, cRGD-PEGDA is an attractive candidate for the future development of tissue engineering scaffolds with optimum cell adhesive strength and ligand density. PMID- 19191568 TI - Influence of disulfide density and molecular weight on disulfide cross-linked polyethylenimine as gene vectors. AB - Disulfide cross-linked polyethylenimines (PEI(X)-SS(Y), where X refers to the molecular weight of raw PEI, and Y refers to the thiolation degree) were prepared in two steps: First, thiol groups were introduced on a raw polyethylenimine (PEI) by the amine-induced ring-opening reaction of thiirane. Second, thiol groups were oxidized by DMSO to form the disulfide cross-links. The cross-linked PEI(800) SS(Y) polymers with a moderate thiolation degree (PEI(800)-SS(2.6,) PEI(800) SS(3.5), and PEI(800)-SS(4.5)) could form compact polyplexes with a size of 200 300 nm at an adequate N/P ratio. In contrast, those with a too low or too high thiolation degree (Y below 2.6 or above 4.5) formed much looser polyplexes with a size above 600 nm. The polyplexes of PEI(X)-SS(3.0-4.0) series (X = 800, 1800, and 25,000) formed small particles with a size below 400 nm at a wide range of N/P ratios. Efficiency of the cross-linked PEIs as gene vectors was evaluated in vitro by transfection of pGL3 to HeLa, COS7, 293T, and CHO cells. The efficiency is disulfide content and molecular weight dependent. The PEI(800)-SS(Y) series with an adequate thiolation degree between 2.6 and 4.5 have relatively lower cytotoxicity and higher gene transfection efficiency than 25 KDa PEI. The polymers with very low or very high thiolation degrees were unable to form compact polyplexes and had very poor transfection efficiency. A suitable molecular weight of raw PEI is also essential to obtain a highly efficient disulfide cross-linked PEI gene vector. Among the three raw PEIs of different molecular weights tested (800 Da, 1800 Da, and 25 KDa), the cross-linked polymer prepared from 800 Da PEI that has the lowest molecular weight gave the best results. PMID- 19191569 TI - Simulation of the first hydration shell of nucleosides D4T and thymidine: structures obtained using MP2 and DFT methods. AB - A comparative theoretical analysis on the effect of the solvent on the molecular structure and energetics of the most stable conformers of the nucleoside analogue D4T (stavudine) and of the natural nucleoside thymidine (Thy) was carried out. Solvent effects were considered using the Tomasi's polarized continuum model (PCM) and including a variable number (1-13) of explicit water molecules surrounding the nucleoside in order to simulate the first hydration shell. More than 200 cluster structures with water were analyzed. B3LYP and MP2 quantum chemical methods were used. The CP-corrected interaction energies for D4T and water molecules were computed. For cases where literature data are available, the computed values were in good agreement with previous experimental and theoretical studies. In the isolated state, conformer I (anti-gg-gg) appears the most stable for D4T molecule and conformer II (anti-gg-gt) for Thy molecule. In D4T with eight water molecules, conformer II changes to conformer I. Thus, conformer I seems preferred when water molecules are situated in the first hydration shell. However, in hydrated thymidine conformer Ia (anti-gg-tg) is the more stable one, and the first hydration shell is more extended than in D4T molecule. The effect of the hydration on the total atomic charges and intermolecular distances were also discussed. Several general conclusions on hydrogen bonds network and involved interaction energies were underlined. PMID- 19191570 TI - Differentiation of rodent bone marrow mesenchymal stem cells into intervertebral disc-like cells following coculture with rat disc tissue. AB - This study aimed to evaluate whether rat mesenchymal stem cells (rMSCs) could be differentiated in vitro into disc-like cells by coculturing with intervertebral disc tissue. rMSCs were cultured with rodent intervertebral disc for up to 30 days in transwell plates. The differentiation of rMSCs was evaluated by immunostaining, Western blot, real-time RT-PCR, Northern blot, and electron microscopy. The potentials of multilineage differentiation and proteoglycan and collagen synthesis were also investigated. rMSCs underwent morphological changes to form three-dimensional micromasses and expressed collagen-2, aggrecan, and sox 9 at RNA and protein levels after 14 days of coculture. These changes were not detected in the samples of rMSCs cultured alone. Cocultured rMSCs also showed other characteristic features of disc-like cells, including the extracellular matrix formation, and proteoglycan and collagen synthesis. In addition, cellular contact between cocultured rMSCs and disc tissue was observed by electron microscopy. Committed rMSCs still retained their differentiation ability into mesoderm lineages of adipocytes or osteocytes when the local environment was altered. This study supports that MSCs are a promising source for cell therapy and tissue engineering in disc regeneration, and highlights that rMSCs can be induced into nucleus pulposus-like cells in vitro under the direct influence of intact disc tissue. PMID- 19191573 TI - Synthesis of benzodiazepine beta-turn mimetics by an Ugi 4CC/Staudinger/aza Wittig sequence. Solving the conformational behavior of the Ugi 4CC adducts. AB - 5-Oxobenzo[e][1,4]diazepine-3-carboxamides were synthesized by sequential Ugi reaction-Staudinger/aza-Wittig cyclization. The pseudopeptidic backbone of the new benzodiazepine derivatives superimposed well with type I, I', II, and II' beta-turn motifs. The intermediate Ugi adducts were characterized as two conformers of the enol form by the correlation between (1)H NMR spectra and X-ray diffraction structures of model compounds. PMID- 19191574 TI - The chemistry of biotic interactions in perspective: small molecules take center stage. AB - The important discoveries made during the last two centuries which are largely responsible for our current understanding of organic chemistry in general, and of natural products chemistry and chemical ecology in particular, are reviewed. A brief personal history follows, including an account of a few examples from our own work which illustrate the importance of interdisciplinary, collaborative research in gaining insights that are not likely to have been achieved by either a chemist or a biologist working alone. Some possible future developments in natural products chemistry and chemical ecology, assuming that we can mobilize appropriate support and enthusiasm for these disciplines, are imagined. Finally, friends, teachers, colleagues, and students who have contributed most importantly to the author's scientific development or who have served as sources of inspiration are gratefully acknowledged. PMID- 19191575 TI - Synthesis of a 35-member stereoisomer library of bistramide A: evaluation of effects on actin state, cell cycle and tumor cell growth. AB - Synthesis and preliminary biological evaluation of a 35-member library of bistramide A stereoisomers are reported. All eight stereoisomers of the C1-C13 tetrahydropyran fragment of the molecule were prepared utilizing crotylsilane reagents 9 and 10 in our [4+2]-annulation methodology. In addition, the four isomers of the C14-C18 gamma-amino acid unit were accessed via a Lewis acid mediated crotylation reaction with use of both enantiomers of organosilane 11. The spiroketal subunit of bistramide A was modified at the C39-alcohol to give another point of stereochemical diversification. The fragments were coupled by using a standard peptide coupling protocol to provide 35 stereoisomers of the natural product. These stereochemical analogues were screened for their effects on cellular actin and cytotoxicity against cancer cell lines (UO-31 renal and SF 295 CNS). The results of these assays identified one analogue, 1.21, with enhanced potency relative to the natural product, bistramide A. PMID- 19191578 TI - Helical-screw directions of diastereoisomeric cyclic alpha-amino acid oligomers. AB - Two series of homooligomers composed of diastereoisomeric cyclic alpha-amino acids having two chiral centers at the alpha-carbon and the side chain were synthesized, and their preferred secondary structures were studied in solution and in the crystal state. The oligomers are a new class of helical-foldamers possessing two kinds of chiral centers on the helical backbone and at the lateral surface of the helix. PMID- 19191576 TI - Proteolytic cleavage of p70 ribosomal S6 kinase by caspase-3 during DNA damage induced apoptosis. AB - p70 S6 kinase (p70S6K) plays an important role in protein translation and cell cycle progression. Increased levels of p70S6K have been associated with drug resistance. In this study, we have investigated the involvement of p70S6K in DNA damage-induced apoptosis. The DNA-damaging agent cisplatin caused a concentration dependent decrease in the level of full-length p70S6K in small cell lung cancer H69 and non-small cell lung cancer A549 cells with a concomitant increase in the level of an approximately 45 kDa fragment. The proteolytic cleavage of p70S6K was inhibited by a broad specificity caspase inhibitor but not by the proteosome or calpain inhibitor. Cell-permeable peptide inhibitor and siRNA against caspase-3 inhibited cisplatin-induced proteolytic cleavage of p70S6K. In vitro-translated p70S6K was cleaved by human recombinant caspase-3. Cisplatin failed to induce cleavage of p70S6K in MCF-7 cells that lack functional caspase-3, but ectopic expression of caspase-3 in MCF-7 cells resulted in the cleavage of p70S6K. p70S6K was primarily cleaved at a noncanonical recognition site, Thr-Pro-Val-Asp, after Asp-393. Site-directed mutagenesis of Asp-393 to Ala resulted in protection against cisplatin-mediated apoptosis, whereas introduction of the N-terminal cleaved fragment resulted in potentiation of cisplatin-induced apoptosis. These results suggest that p70S6K is a novel substrate for caspase-3 and that the proteolytic cleavage of p70S6K is important for cisplatin-induced apoptosis. PMID- 19191580 TI - Tandem ring-opening/closing reactions of N-Ts aziridines and aryl propargyl alcohols promoted by t-BuOK. AB - t-BuOK was found to be an effective promoting reagent for tandem ring opening/closing reactions of various N-Ts aziridines and aryl propargyl alcohols to afford dihydroxazine derivatives in moderate to good yields. A plausible reaction mechanism has been proposed. PMID- 19191579 TI - Targeted gene delivery mediated by folate-polyethylenimine-block-poly(ethylene glycol) with receptor selectivity. AB - The folate receptor (FR) is a tumor marker overexpressed in large numbers of cancer cells. Folic acid has high affinity to the FR and retains its binding affinity upon derivatization via its gamma-carboxyl. Therefore, in this article, folate-polyethylenimine-block-poly(ethylene glycol) (FOL-PEI-b-PEG) was designed for specific receptor targeted gene delivery. Physicochemical characterizations of resulting FOL-PEI-b-PEG/DNA complexes in terms of agarose gel electrophoresis, particle size, and zeta potential measurements were investigated. The results indicated that FOL-PEI-b-PEG was able to condense plasmid DNA tightly with a suitable particle size. The cytotoxicity study indicated that the copolymer exhibited less toxicity in comparison with that of 25 kDa PEI. Luciferase assay and green fluorescent protein (GFP) detections were also used to confirm that FOL PEI-b-PEG could be an effective gene vector. Importantly, transfection efficiency of FOL-PEI-b-PEG with free folic acid was much lower than that of the copolymer without free folic acid on FR-positive HeLa cells, suggesting that FOL-PEI-b-PEG has great potential as a targeting gene vector. PMID- 19191581 TI - Quantitation of twelve metals in tequila and mezcal spirits as authenticity parameters. AB - In this paper the differentiation of silver, gold, aged and extra-aged tequila and mezcal has been carried out according to their metal content. Aluminum, barium, calcium, copper, iron, magnesium, manganese, potassium, sodium, strontium, zinc, and sulfur were determined by inductively coupled plasma optical emission spectrometry. The concentrations found for each element in the samples were used as chemical descriptors for characterization purposes. Principal component analysis, linear discriminant analysis and artificial neural networks were applied to differentiate types of tequila and mezcal. Using probabilistic neural networks 100% of success in the classification was obtained for silver, gold, extra-aged tequila and mezcal. In the case of aged tequila 90% of samples were successfully classified. Sodium, potassium, calcium, sulfur, magnesium, iron, strontium, copper and zinc were the most discriminant elements. PMID- 19191582 TI - PNA-PEG modified silicon platforms as functional bio-interfaces for applications in DNA microarrays and biosensors. AB - The synthesis and characterization of two types of silicon-based biofunctional interfaces are reported; each interface bonds a dense layer of poly(ethylene glycol) (PEG(n)) and peptide nucleic acid (PNA) probes. Phosphonate self assembled monolayers were derivatized with PNA using a maleimido-terminated PEG(45). Similarly, siloxane monolayers were functionalized with PNA using a maleimido-terminated PEG(45) spacer and were subsequently modified with a shorter methoxy-terminated PEG(12) ("back-filling"). The long PEG(45) spacer was used to distance the PNA probe from the surface and to minimize undesirable nonspecific adsorption of DNA analyte. The short PEG(12) "back-filler" was used to provide additional passivation of the surface against nonspecific DNA adsorption. X-ray photoelectron spectroscopic (XPS) analysis near the C 1s and N 1s ionization edges was done to characterize chemical groups formed in the near-surface region, which confirmed binding of PEG and PNA to the phosphonate and silane films. XPS also indicated that additional PEG chains were tethered to the surface during the back-filling process. Fluorescence hybridization experiments were carried out with complementary and noncDNA strands; both phosphonate and siloxane biofunctional surfaces were effective for hybridization of cDNA strands and significantly reduced nonspecific adsorption of the analyte. Spatial patterns were prepared by polydimethylsiloxane (PDMS) micromolding on the PNA functionalized surfaces; selective hybridization of fluorescently labeled DNA was shown at the PNA functionalized regions, and physisorption at the probe-less PEG functionalized regions was dramatically reduced. These results show that PNA-PEG derivatized phosphonate monolayers hold promise for the smooth integration of device surface chemistry with semiconductor technology for the fabrication of DNA biosensors. In addition, our results confirm that PNA-PEG derivatized self assembled carboxyalkylsiloxane films are promising substrates for DNA microarray applications. PMID- 19191583 TI - Efficient kinetic macrocyclization. AB - In this article, the highly efficient formation of a series of recently discovered aromatic oligoamide macrocycles consisting of six meta-linked residues is first discussed. The macrocycles, with their backbones rigidified by three center hydrogen bonds, were found to form in high yields that deviate dramatically from the theoretically allowed value obtained from kinetic simulation of a typical kinetically controlled macrocyclization reaction. The folding of the uncyclized six-residue oligomeric precursors, which belong to a class of backbone-rigidified oligoamides that have been demonstrated by us to adopt well-defined crescent conformations, plays a critical role in the observed high efficiency. Out of two possible mechanisms, one is consistent with experimental results obtained from the coupling of crescent oligoamides of different lengths, which suggests a remote steric effect that discourages the formation of oligomers having lengths longer than the backbone of the six-residue precursors. The suggested mechanism is supported by the efficient formation of very large aromatic oligoamide macrocycles consisting of alternating meta- and para-linked residues. These large macrocycles, having H-bond-rigidified backbones and large internal lumens, are formed in high (>80%) yields on the basis of one step, multicomponent macrocyclization reactions. The condensation of monomeric meta-diamines and a para-diacid chloride leads to the efficient formation of macrocycles with 14, 16, and 18 residues, corresponding to 70-, 80-, and 90 membered rings that contain internal cavities of 2.2, 2.5, and 2.9 nm across. In addition, the condensation between trimeric or pentameric diamines and a monomeric diacid chloride had resulted in the selective formation of single macrocyclic products with 16 or 18 residues. The efficient formation of the macrocycles, along with the absence of other noncyclic oligomeric and polymeric byproducts, is in sharp contrast to the poor yields associated with most kinetically controlled macrocyclization reactions. This system represents a rare example of highly efficient kinetic macrocyclization reactions involving large numbers of reacting units, which provides very large, shape-persistent macrocycles. PMID- 19191584 TI - Efficient chemical synthesis of AppDNA by adenylation of immobilized DNA-5' monophosphate. AB - AppDNA is an intermediate in enzyme-catalyzed DNA ligation reactions, and its efficient enzymatic synthesis requires a donor-template duplex of at least 11 base pairs in length. An efficient chemical synthesis of AppDNA with the coupling of an adenosine 5'-phosphorimidazolidate to an immobilized DNA-5'-monophosphate as the key step is described. The adenylation efficiencies of DNA-5' monophosphate were excellent for oligonucleotides containing less than 11 nucleotides and at least 50% for oligonucleotides containing 15-25 nucleotides. PMID- 19191585 TI - Antagonists of the P2X(7) receptor. From lead identification to drug development. PMID- 19191587 TI - Disruption of human plasma high-density lipoproteins by streptococcal serum opacity factor requires labile apolipoprotein A-I. AB - Human plasma high-density lipoproteins (HDL), the primary vehicle for reverse cholesterol transport, are the target of serum opacity factor (SOF), a virulence determinant of Streptococcus pyogenes that turns serum opaque. HDL comprise a core of neutral lipidscholesteryl esters and some triglyceridesurrounded by a surface monolayer of cholesterol, phospholipids, and specialized proteins [apolipoproteins (apos) A-I and A-II]. A HDL is an unstable particle residing in a kinetic trap from which it can escape via chaotropic, detergent, or thermal perturbation. Recombinant (r) SOF catalyzes the transfer of nearly all neutral lipids of approximately 100,000 HDL particles (D approximately 8.5 nm) into a single, large cholesteryl ester-rich microemulsion (CERM; D > 100 nm), leaving a new HDL-like particle [neo HDL (D approximately 5.8 nm)] while releasing lipid free (LF) apo A-I. CERM formation and apo A-I release have similar kinetics, suggesting parallel or rapid consecutive steps. By using complementary physicochemical methods, we have refined the mechanistic model for HDL opacification. According to size exclusion chromatography, a HDL containing nonlabile apo A-I resists rSOF-mediated opacification. On the basis of kinetic cryo-electron microscopy, rSOF (10 nM) catalyzes the conversion of HDL (4 microM) to neo HDL via a stepwise mechanism in which intermediate-sized particles are seen. Kinetic turbidimetry revealed opacification as a rising exponential reaction with a rate constant k of (4.400 +/- 0.004) x 10(-2) min(-1). Analysis of the kinetic data using transition state theory gave an enthalpy (DeltaH()), entropy (DeltaS(++)), and free energy (DeltaG()) of activation of 73.9 kJ/mol, 66.87 J/K, and 94.6 kJ/mol, respectively. The free energy of activation for opacification is nearly identical to that for the displacement of apo A-I from HDL by guanidine hydrochloride. We conclude that apo A-I lability is required for HDL opacification, LF apo A-I desorption is the rate-limiting step, and nearly all HDL particles contain at least one labile copy of apo A-I. PMID- 19191588 TI - Chromophore-assisted laser inactivation of alpha- and gamma-tubulin SNAP-tag fusion proteins inside living cells. AB - Chromophore-assisted laser inactivation (CALI) can help to unravel localized activities of target proteins at defined times and locations within living cells. Covalent SNAP-tag labeling of fusion proteins with fluorophores such as fluorescein is a fast and highly specific tool to attach the photosensitizer to its target protein in vivo for selective inactivation of the fusion protein. Here, we demonstrate the effectiveness and specificity of SNAP-tag-based CALI by acute inactivation of alpha-tubulin and gamma-tubulin SNAP-tag fusions during live imaging assays of cell division. Singlet oxygen is confirmed as the reactive oxygen species that leads to loss of fusion protein function. The major advantage of SNAP-tag CALI is the ease, reliability, and high flexibility in labeling: the genetically encoded protein tag can be covalently labeled with various dyes matching the experimental requirements. This makes SNAP-tag CALI a very useful tool for rapid inactivation of tagged proteins in living cells. PMID- 19191586 TI - Targeting the fatty acid biosynthesis enzyme, beta-ketoacyl-acyl carrier protein synthase III (PfKASIII), in the identification of novel antimalarial agents. AB - The importance of fatty acids to the human malaria parasite, Plasmodium falciparum, and differences due to a type I fatty acid synthesis (FAS) pathway in the parasite, make it an attractive drug target. In the present study, we developed and a utilized a pharmacophore to select compounds for testing against PfKASIII, the initiating enzyme of FAS. This effort identified several PfKASIII inhibitors that grouped into various chemical classes of sulfides, sulfonamides, and sulfonyls. Approximately 60% of the submicromolar inhibitors of PfKASIII inhibited in vitro growth of the malaria parasite. These compounds inhibited both drug sensitive and resistant parasites and testing against a mammalian cell line revealed an encouraging in vitro therapeutic index for the most active compounds. Docking studies into the active site of PfKASIII suggest a potential binding mode that exploits amino acid residues at the mouth of the substrate tunnel. PMID- 19191592 TI - Capacity of a single spiking neuron channel. AB - Information transfer through a single neuron is a fundamental component of information processing in the brain, and computing the information channel capacity is important to understand this information processing. The problem is difficult since the capacity depends on coding, characteristics of the communication channel, and optimization over input distributions, among other issues. In this letter, we consider two models. The temporal coding model of a neuron as a communication channel assumes the output is tau where tau is a gamma distributed random variable corresponding to the interspike interval, that is, the time it takes for the neuron to fire once. The rate coding model is similar; the output is the actual rate of firing over a fixed period of time. Theoretical studies prove that the distribution of inputs, which achieves channel capacity, is a discrete distribution with finite mass points for temporal and rate coding under a reasonable assumption. This allows us to compute numerically the capacity of a neuron. Numerical results are in a plausible range based on biological evidence to date. PMID- 19191590 TI - Regimen simplification to atazanavir-ritonavir alone as maintenance antiretroviral therapy: final 48-week clinical and virologic outcomes. AB - BACKGROUND: Simplified maintenance therapy with ritonavir-boosted atazanavir (ATV/RTV) alone is attractive because of nucleoside reverse-transcriptase inhibitor (NRTI)-sparing benefits, low pill burden, once-daily dosage, and safety. METHODS: Subjects with virologic suppression after > or = 48 weeks of initial antiretroviral therapy with 2 NRTIs and a protease inhibitor (PI) were enrolled. Subjects switched to ATV/RTV at entry and discontinued NRTIs after 6 weeks. The primary end point was time to virologic failure (confirmed HIV-1 RNA level > or = 200 copies/mL). Drug resistance at virologic failure was evaluated by standard genotyping and single-genome sequencing (SGS). Residual viremia (1.1 49 copies/mL) was measured by single-copy assay. RESULTS: Thirty-four subjects simplified to ATV/RTV alone, of whom 30 (88%) did not experience virologic failure by 48 weeks after simplification. Residual viremia did not change significantly after NRTI discontinuation among those without virologic failure but did increase 4-12 weeks before confirmed virologic failure. No major PI resistance mutations were identified at virologic failure by standard genotyping or SGS. CONCLUSIONS: In this pilot study, simplified maintenance therapy with ATV/RTV alone maintained viral suppression in most subjects through 48 weeks. PI resistance was not detected among subjects experiencing virologic failure. Larger, randomized trials are warranted to further define the efficacy and safety of this strategy. PMID- 19191593 TI - A binary variable model for affinity propagation. AB - Affinity propagation (AP) was recently introduced as an unsupervised learning algorithm for exemplar-based clustering. We present a derivation of AP that is much simpler than the original one and is based on a quite different graphical model. The new model allows easy derivations of message updates for extensions and modifications of the standard AP algorithm. We demonstrate this by adjusting the new AP model to represent the capacitated clustering problem. For those wishing to investigate or extend the graphical model of the AP algorithm, we suggest using this new formulation since it allows a simpler and more intuitive model manipulation. PMID- 19191594 TI - Estimation of positive semidefinite correlation matrices by using convex quadratic semidefinite programming. AB - The correlation matrix is a fundamental statistic that is used in many fields. For example, GroupLens, a collaborative filtering system, uses the correlation between users for predictive purposes. Since the correlation is a natural similarity measure between users, the correlation matrix may be used in the Gram matrix in kernel methods. However, the estimated correlation matrix sometimes has a serious defect: although the correlation matrix is originally positive semidefinite, the estimated one may not be positive semidefinite when not all ratings are observed. To obtain a positive semidefinite correlation matrix, the nearest correlation matrix problem has recently been studied in the fields of numerical analysis and optimization. However, statistical properties are not explicitly used in such studies. To obtain a positive semidefinite correlation matrix, we assume the approximate model. By using the model, an estimate is obtained as the optimal point of an optimization problem formulated with information on the variances of the estimated correlation coefficients. The problem is solved by a convex quadratic semidefinite program. A penalized likelihood approach is also examined. The MovieLens data set is used to test our approach. PMID- 19191595 TI - Dynamical analysis of Bayesian inference models for the Eriksen task. AB - The Eriksen task is a classical paradigm that explores the effects of competing sensory inputs on response tendencies and the nature of selective attention in controlling these processes. In this task, conflicting flanker stimuli interfere with the processing of a central target, especially on short reaction time trials. This task has been modeled by neural networks and more recently by a normative Bayesian account. Here, we analyze the dynamics of the Bayesian models, which are nonlinear, coupled discrete time dynamical systems, by considering simplified, approximate systems that are linear and decoupled. Analytical solutions of these allow us to describe how posterior probabilities and psychometric functions depend on model parameters. We compare our results with numerical simulations of the original models and derive fits to experimental data, showing that agreements are rather good. We also investigate continuum limits of these simplified dynamical systems and demonstrate that Bayesian updating is closely related to a drift-diffusion process, whose implementation in neural network models has been extensively studied. This provides insight into how neural substrates can implement Bayesian computations. PMID- 19191596 TI - Generation of spike trains with controlled auto- and cross-correlation functions. AB - Emerging evidence indicates that information processing, as well as learning and memory processes, in both the network and single-neuron levels are highly dependent on the correlation structure of multiple spike trains. Contemporary experimental as well as theoretical studies that involve quasi-realistic neuronal stimulation thus require a method for controlling spike train correlations. This letter introduces a general new strategy for generating multiple spike trains with exactly controlled mean firing rates and correlation structure (defined in terms of auto- and cross-correlation functions). Our approach nonlinearly transforms random gaussian-distributed processes with a predistorted correlation structure into nonnegative rate processes, which are then used to generate doubly stochastic Poisson point processes with the required correlation structure. We show how this approach can be used to generate stationary or nonstationary spike trains from small or large groups of neurons with diverse auto- and cross correlation structures. We analyze and derive analytical formulas for the high order correlation structure of generated spike trains and discuss the limitations of this approach. PMID- 19191597 TI - Adaptive synchronization of activities in a recurrent network. AB - Predictive learning rules, where synaptic changes are driven by the difference between a random input and its reconstruction derived from internal variables, have proven to be very stable and efficient. However, it is not clear how such learning rules could take place in biological synapses. Here we propose an implementation that exploits the synchronization of neural activities within a recurrent network. In this framework, the asymmetric shape of spike-timing dependent plasticity (STDP) can be interpreted as a self-stabilizing mechanism. Our results suggest a novel hypothesis concerning the computational role of neural synchrony and oscillations. PMID- 19191598 TI - How does maintenance of network activity depend on endogenous dynamics of isolated neurons? AB - Robust activity of some networks, such as central pattern generators, suggests the existence of physiological mechanisms that maintain the most important characteristics, for example, the period and spike frequency of the pattern. Whatever these mechanisms are, they change the appropriate model parameters to or along the isomanifolds on which the characteristics of the pattern are constant, while their sensitivities to parameters may be different. Setting synaptic connections to zero at the points of isomanifolds allows for dissecting the maintenance mechanisms into components involving synaptic transmission and components involving intrinsic currents. The physiological meaning of the intrinsic current changes might be revealed by analysis of their impact on endogenous neuronal dynamics. Here, we sought answers to two questions: (1) Do parameter variations in insensitive directions (along isomanifolds) change endogenous dynamics of the network neurons? (2) Do sensitive and insensitive directions for network pattern characteristics depend on endogenous dynamics of the network neurons? We considered a leech heartbeat half-center oscillator model network and analyzed isomanifolds on which the burst period or spike frequency of the model, or both, are constant. Based on our analysis, we hypothesize that the dependence on endogenous dynamics of the isolated neurons is the stronger the more characteristics of the pattern have to be maintained. We also found that in general, the network was more flexible when it consisted of endogenously tonically spiking rather than bursting or silent neurons. Finally, we discuss the physiological implications of our findings. PMID- 19191599 TI - Nonlinear extraction of independent components of natural images using radial gaussianization. AB - We consider the problem of efficiently encoding a signal by transforming it to a new representation whose components are statistically independent. A widely studied linear solution, known as independent component analysis (ICA), exists for the case when the signal is generated as a linear transformation of independent nongaussian sources. Here, we examine a complementary case, in which the source is nongaussian and elliptically symmetric. In this case, no invertible linear transform suffices to decompose the signal into independent components, but we show that a simple nonlinear transformation, which we call radial gaussianization (RG), is able to remove all dependencies. We then examine this methodology in the context of natural image statistics. We first show that distributions of spatially proximal bandpass filter responses are better described as elliptical than as linearly transformed independent sources. Consistent with this, we demonstrate that the reduction in dependency achieved by applying RG to either nearby pairs or blocks of bandpass filter responses is significantly greater than that achieved by ICA. Finally, we show that the RG transformation may be closely approximated by divisive normalization, which has been used to model the nonlinear response properties of visual neurons. PMID- 19191601 TI - GABA transporter preserving ongoing spontaneous neuronal activity at firing subthreshold. AB - There has been compelling evidence that the GABA transporter is crucial not only for removing gamma-aminobutyric acid (GABA) from but also releasing it into extracellular space, thereby clamping ambient GABA (GABA in extracellular space) at a certain level. The ambient GABA is known to activate extrasynaptic GABA receptors and provide tonic inhibitory current into neurons. We investigated how the transporter regulates the level of ambient GABA, mediates tonic neuronal inhibition, and influences ongoing spontaneous neuronal activity. A cortical neural network model is proposed in which GABA transporters on lateral (L) and feedback (F) inhibitory (GABAergic) interneurons are functionally made. Principal (P) cell assemblies participate in expressing information about elemental sensory features. At membrane potentials below the reversal potential, there is net influx of GABA, whereas at membrane potentials above the reversal potential, there is net efflux of GABA. Through this transport mechanism, ambient GABA concentration is kept within a submicromolar range during an ongoing spontaneous neuronal activity time period. Here we show that the GABA transporter on L cells regulates the overall level of ambient GABA across cell assemblies, and that on F cells it does so within individual cell assemblies. This combinatorial regulation of ambient GABA allows P cells to oscillate near firing threshold during the ongoing time period, thereby reducing their reaction time to externally applied stimuli. We suggest that the GABA transporter, with its forward and reverse transport mechanism, could regulate the ambient GABA. This transporter-mediated ambient GABA regulation may contribute to establishing an ongoing subthreshold neuronal state by which the network can respond rapidly to subsequent sensory input. PMID- 19191600 TI - A first-order nonhomogeneous Markov model for the response of spiking neurons stimulated by small phase-continuous signals. AB - We present a first-order nonhomogeneous Markov model for the interspike-interval density of a continuously stimulated spiking neuron. The model allows the conditional interspike-interval density and the stationary interspike-interval density to be expressed as products of two separate functions, one of which describes only the neuron characteristics and the other of which describes only the signal characteristics. The approximation shows particularly clearly that signal autocorrelations and cross-correlations arise as natural features of the interspike-interval density and are particularly clear for small signals and moderate noise. We show that this model simplifies the design of spiking neuron cross-correlation systems and describe a four-neuron mutual inhibition network that generates a cross-correlation output for two input signals. PMID- 19191602 TI - A new hypothesis for sleep: tuning for criticality. AB - We propose that the critical function of sleep is to prevent uncontrolled neuronal feedback while allowing rapid responses and prolonged retention of short term memories. Through learning, the brain is tuned to react optimally to environmental challenges. Optimal behavior often requires rapid responses and the prolonged retention of short-term memories. At a neuronal level, these correspond to recurrent activity in local networks. Unfortunately, when a network exhibits recurrent activity, small changes in the parameters or conditions can lead to runaway oscillations. Thus, the very changes that improve the processing performance of the network can put it at risk of runaway oscillation. To prevent this, stimulus-dependent network changes should be permitted only when there is a margin of safety around the current network parameters. We propose that the essential role of sleep is to establish this margin by exposing the network to a variety of inputs, monitoring for erratic behavior, and adjusting the parameters. When sleep is not possible, an emergency mechanism must come into play, preventing runaway behavior at the expense of processing efficiency. This is tiredness. PMID- 19191603 TI - Multifrequency electron spin resonance spectra of a spin-labeled protein calculated from molecular dynamics simulations. AB - Multifrequency electron spin resonance (ESR) spectra provide a wealth of structural and dynamic information about the local environment of the spin label and, indirectly, about the spin-labeled protein. Relating the features of the observed spectra to the underlying molecular motions and interactions is, however, challenging. To make progress toward a rigorous interpretation of ESR spectra, we perform extensive molecular dynamics (MD) simulations of fully solvated T4 Lysozyme, labeled with the spin label MTSSL at positions 72 and 131. These two sites have been the object of numerous experimental studies and are generally considered as prototypical solvent-exposed sites on the surfaces of alpha-helices. To extend the time window afforded by the MD simulations, stochastic Markov models reflecting the dynamics of the spin label side chains in terms of their rotameric states are constructed from the trajectories. The calculated multifrequency ESR spectra are in very good agreement with experiment for three different magnetic field strengths without adjusting any parameters. During the trajectories, the spin labels interconvert among a fairly large number of conformations and display a propensity to form interactions with protein residues other than their nearest neighbors along the helix. The detailed picture of the spin label emerging from the MD simulations provides useful insight into the molecular origins of the available spectroscopic and crystallographic data. PMID- 19191604 TI - Balancing adherence concerns with the risks of HIV disease progression. PMID- 19191605 TI - Invasive pneumococcal disease among children in rural Bangladesh: results from a population-based surveillance. AB - BACKGROUND: Streptococcus pneumoniae infection is recognized as a global priority public health problem, and conjugate vaccines have been shown to prevent vaccine type invasive pneumococcal disease (IPD) in children. However, better estimates of the disease burden and reliable population-based data on serotype composition are needed for vaccine development and implementation in developing countries. METHODS: We initiated a population-based surveillance in the rural Bangladesh community of Mirzapur, covering a population of approximately 144,000. Village health care workers made weekly visits to approximately 12,000 children 1-59 months of age in the study area. Children with reported fever, cough, or difficulty breathing were assessed by the village health care workers using a clinical algorithm and were referred to the hospital if required. Children from the study area who were seen in the hospital underwent clinical examination and laboratory testing if they met standardized case definitions. IPD was confirmed by blood and/or cerebrospinal fluid culture results. Isolates were identified, tested for susceptibility to antibiotics, and serotyped in accordance with standard laboratory methods. We present here the results from the first 3 years of the surveillance (July 2004-June 2007). RESULTS: Village health care workers identified 5020 cases of possible severe pneumonia and/or very severe disease (165 cases per 1000 child-years)and 9411 cases of possible pneumonia (310 cases per 1000 child-years) as well as 2029 cases of suspected meningitis and/or very severe disease (67 cases per 1000 child-years) and 8967 cases of high fever and/or possible bacteremia (295 cases per 1000 child-years). Pneumonia was the single most common form of illness observed among 2596 hospitalizations (found in 977 [38%] of cases). We recovered 26 S. pneumoniae isolates (25 isolates from 6925 blood cultures and 1 isolate from 41 cerebrospinal fluid cultures), which gave an overall IPD incidence of 86 cases per 100,000 child-years. Invasive pneumococcal infection was common during infancy (with infants accounting for 23 of the 26 cases), and 50% of the total isolates were obtained from nonhospitalized patients who received a diagnosis of upper respiratory tract infection and fever. The most prevalent pneumococcal serotypes were serotypes 1, 5, 14, 18C, 19A, and 38. Ten of the 26 isolates were completely resistant to trimethoprim-sulfamethoxazole, and another 10 isolates had intermediate resistance. CONCLUSIONS: IPD contributes substantially to childhood morbidity in rural Bangladesh. S. pneumoniae can cause invasive but nonsevere disease in children, and IPD incidence can be seriously under reported if such cases are overlooked. The emerging high resistance to trimethoprim-sulfamethoxazole should be addressed. Data on serotype distribution would help to guide appropriate pneumococcal conjugate vaccine formulation. PMID- 19191606 TI - Hospital-based surveillance of invasive pneumococcal disease among young children in urban Nepal. AB - BACKGROUND: Streptococcus pneumoniae is a leading cause of pneumonia and meningitis in young children. Before implementation of the pneumococcal conjugate vaccine in developing countries, there is an urgent need to provide regional epidemiological data on pneumococcal disease. The aims of this study were to determine the prevalence and serotype distribution of invasive pneumococcal disease among young children hospitalized in urban Nepal. METHODS: Children aged 2 months to 5 years who were admitted to Patan Hospital, Kathmandu, with fever and/or suspected pneumonia, meningitis, or bacteremia were recruited. Blood culture specimens were collected from all participants. In cases of suspected meningitis, cerebrospinal fluid specimens were cultured and were tested for S. pneumoniae antigen. RESULTS: A total of 885 children were recruited during the 21 month study period. Of these, 76 (9%) had meningitis and 498 (56%) had pneumonia, on the basis of clinical criteria. Radiographically confirmed pneumonia occurred in 354 (40%), and probable or definite meningitis occurred in 47 (5%). S. pneumoniae was isolated in specimens from 17 (2%) of the children. Serotypes 1 and 12A were isolated most frequently, and only 1 of 17 isolates had a serotype contained in the currently available 7-valent pneumococcal conjugate vaccine. CONCLUSIONS: More than 60% of children aged <5 years who were admitted with fever and/or suspected invasive bacterial disease in urban Nepal had the clinical syndromes of meningitis and/or pneumonia. A new generation of pneumococcal vaccines that prevent infection with a broader range of serotypes may be necessary to most effectively control pneumococcal disease in young children in Kathmandu. PMID- 19191607 TI - Invasive pneumococcal disease in Kanti Children's Hospital, Nepal, as observed by the South Asian Pneumococcal Alliance network. AB - BACKGROUND: Pneumonia accounts for approximately 2 million deaths annually among children aged <5 years, with most of these deaths occurring in Africa and southern Asia. The South Asian Pneumococcal Alliance (SAPNA) network in Nepal is generating local epidemiological data to assist in the development of national and regional policies for prevention of pneumococcal and Haemophilus influenzae (Hib) disease. METHODS: Children aged 2 months to 5 years with suspected invasive bacterial disease were recruited from Kanti Children Hospital, Kathmandu, Nepal. Specimens of blood, CSF, and normally sterile body fluids were cultured, and analysis of antimicrobial susceptibility patterns and serotyping of Streptococcus pneumoniae isolates were performed. CSF specimens were also tested for S. pneumoniae and Hib antigens by a latex agglutination test and an immunochromatographic test of pneumococcal antigen (NOW S. pneumoniae Antigen Test; Binax). RESULTS: A total of 2528 children with suspected invasive bacterial disease were recruited, of whom 82% had pneumonia, 9.6% had meningitis, 2% had very severe disease, and 0.4% had bacteremia; the remainder received another diagnosis. Before hospitalization, 26.7% had received antibiotic treatment. Fifty children had S. pneumoniae identified as the etiological agent of invasive disease. Of 2461 blood cultures performed, 22 were positive for S. pneumoniae. Of 33 cases of S. pneumoniae meningitis, 11 were detected by CSF culture, and 21 were detected by latex agglutination and pneumococcal antigen tests. The rate of detection of S. pneumoniae in CSF was 3.6% by culture, compared with 7.8% by latex agglutination and 10% by pneumococcal antigen testing. The rate of detection of H. influenzae in CSF was 1.7% by culture and 6.5% by latex agglutination. The most common serotypes found were 1, 5, 2, and 7F, followed by 12A, 19B, and 23F. Of all the invasive isolates, 3.8% were resistant to penicillin, and 68% were resistant to trimethoprim-sulfamethoxazole. CONCLUSIONS: The SAPNA network has identified Hib and pneumococci as causes of significant disease in Nepal. PMID- 19191608 TI - Surveillance of pneumococcal meningitis among children in Sindh, southern Pakistan. AB - BACKGROUND: Information about the burden of invasive pneumococcal disease among children in Pakistan is limited. METHODS: Surveillance of bacterial meningitis among children aged <5 years was set up at 18 hospitals in southern Pakistan that fulfilled the following criteria: (1) >30 pediatric admissions weekly, (2) skilled personnel to perform lumbar punctures, and (3) close proximity to an Aga Khan University Hospital laboratory collection point. RESULTS: A total of 2690 children were admitted to the hospital with suspected acute bacterial meningitis, and 2646 (98%) underwent lumbar puncture. Of the 2646 cerebrospinal fluid specimens obtained, 412 (16%) were purulent, and pathogens were detected by culture or latex agglutination testing in 83 (20.1%) of the purulent specimens. Of the 83 isolates detected, 48 (57.8%) were Haemophilus influenzae type b, 32 (38.5%) were Streptococcus pneumoniae, and 3 (3.6%) were Neisseria meningiditis. Overall, 81% of the pathogens detected were from children aged <1 year. More than 50% of families reported definite prior antimicrobial use. The minimum detected incidence rates of purulent meningitis in Hyderabad were 112 cases per 100,000 children aged <1 year and 45.3 cases per 100,000 children aged <5 years. After adjustment for limitations in access to care and the low sensitivity of cerebrospinal fluid culture, the adjusted incidence rates of pneumococcal meningitis were 81 cases per 100,000 children aged <1 year (95% confidence interval, 26.2-190.5 cases per 100,000) and 20 cases per 100,000 children aged <5 years (95% confidence interval, 7.3-43.7 cases per 100,000). Of the 32 children with pneumococcal meningitis, 8 (25%) died during hospitalization. CONCLUSIONS: Our surveillance system detected a substantial burden of purulent meningitis among infants and children in southern Pakistan. H. influenzae type b and S. pneumoniae accounted for >90% of detected pathogens. The use of vaccines against these 2 pathogens could prevent a substantial portion of disease and deaths in Pakistan. PMID- 19191609 TI - Surveillance of invasive pneumococcal disease in Colombo, Sri Lanka. AB - The South Asian Pneumococcal Surveillance network uses standard recruitment and laboratory procedures for surveillance of invasive pneumococcal disease in India, Nepal, and Sri Lanka. Children aged 2 months to 5 years who were admitted to the sentinel surveillance site, Lady Ridgeway Hospital for Children, in Colombo, Sri Lanka, and who presented with signs and symptoms of meningitis, pneumonia, or very severe disease were studied. Blood culture and CSF culture specimens were analyzed at the microbiology laboratory at Lady Ridgeway Hospital for Children. Specimens were processed by routine conventional methods. Antigen testing was performed on CSF specimens with use of commercially available latex agglutination test kits. From January 2005 to March 2007, we observed 23 isolates of Streptococcus pneumoniae, and the most common serotypes were 19F, 14, 23F, and 6B. Of the serotypes found, 60% are covered by the currently available 7-valent conjugate pneumococcal vaccine. More than 90% of the isolates were penicillin resistant, and the rate of resistance to third-generation cephalosporins was also high. PMID- 19191610 TI - Report on invasive disease and meningitis due to Haemophilus influenzae and Streptococcus pneumonia from the Network for Surveillance of Pneumococcal Disease in the East African Region. AB - Pneumococcal disease in young children has not been as well characterized in East Africa as it has been in industrialized countries. Although pneumococci are likely to cause substantial mortality and morbidity, universal diagnostic challenges plus the rudimentary nature of public health surveillance make the true epidemiological characteristics of these diseases difficult to ascertain with these methods alone. However, local data are critical to inform the debate on vaccine deployment and assess vaccine impact. The Network for Surveillance of Pneumococcal Disease in the East African Region has worked to expand the World Health Organization Paediatric Bacterial Meningitis Surveillance Network initiated surveillance process aimed at Haemophilus influenza type b to perform surveillance on pneumococcal diseases. A total of 119 H. influenzae isolates from children aged >2 years but <5 years of age have been confirmed. Eighty-three isolates (69.75%) were serotype b, 19 belonged to other capsular antigen groups, and 17 were nontypable. For Streptococcus pneumoniae, a total of 442 isolates were confirmed to be pneumococci; 302 isolates were from blood cultures, and 140 were from cultures of cerebrospinal fluid. Most of the isolates were obtained from patients in the 6-29-month age group; in this age group, overall coverage by the heptavalent vaccine was 56% (increasing to 67% with the addition of cross protection due to serotype 6A). S. pneumoniae isolates are susceptible to most commonly used antibiotics, with the exception of trimethoprim-sulfamethoxazole, and have exhibited no resistance to penicillin. A surveillance network is in place to provide local data on the importance of S. pneumoniae as a cause of both meningitis and bacteremia. Serotypes in the currently available heptavalent conjugate pneumococcal vaccine and related serotypes account for two-thirds of invasive pneumococcal disease among children aged 6-29 months. PMID- 19191611 TI - Surveillance for Streptococcus pneumoniae meningitis in children aged <5 years: implications for immunization in Uganda. AB - Affordable pneumococcal conjugate vaccines will soon become available to developing countries through the Global Alliance for Vaccines and Immunization. Data on Streptococcus pneumoniae meningitis epidemiology in Uganda will assist decision makers in determining the best national vaccine policy. We reviewed acute bacterial meningitis surveillance data for children aged <5 years from 3 sentinel surveillance sites in 3 Ugandan districts collected from 2001 through 2006. Serotype and antibiotic-resistance testing were performed on pneumococcal isolates collected from 2005 through 2006 from the Kampala district in the tropical central region of Uganda. Minimum pneumococcal meningitis incidence estimates were calculated for a portion of the Kampala district and all of the Gulu district, where case ascertainment was more complete. At the 3 sites, 14,388 probable acute bacterial meningitis cases were observed. The most common cause identified was S. pneumoniae (n = 331; 35% of all confirmed cases), which had an overall case fatality ratio of 19%. Yearly pneumococcal meningitis incidence was 3-20 cases per 100,000 population in Kampala versus 28-42 cases per 100,000 population in Gulu. The most commonly identified serotypes were 6A/6B (40%); 43% of isolates were serotypes that are in the available 7-valent pneumococcal conjugate vaccine and 70% are in the proposed 13-valent pneumococcal vaccine. Twenty-five isolates (83%) had intermediate resistance to penicillin but none were fully resistant. Pneumococcal meningitis is common and severe in Uganda, indicating a role for the pneumococcal conjugate vaccine. PMID- 19191612 TI - Behind the data: establishing the Network for Surveillance of Pneumococcal Disease in the East African Region. AB - In a region with high rates of mortality among children aged <5 years, the underfunded health care systems of sub-Saharan Africa have few resources available to perform surveillance activities that can help determine the causes of morbidity and mortality in the region. At present, there are few examples of attempts to promote public health care surveillance that might inform current debates about how to expand and improve surveillance, particularly for bacterial diseases. Driven by this gap in knowledge, we attempted to explore the successes and failures of the Network for Surveillance of Pneumococcal Disease in the East African Region and to share the experiences of what are essentially nonresearch public-sector hospitals in East Africa, with the hopes that surveillance systems for other diseases, especially those that require complex diagnostic support, may be informed by these experiences. The state of services essential for surveillance and the measures taken to overcome any shortcomings are described, as is the progress made in improving clinical diagnosis, laboratory processing, and data management. For surveillance to play a role in public health care, ministries of health and associated institutions must own and push forward the surveillance agenda, with support from global partners, and take advantage of the developments that have been achieved within the institutions. PMID- 19191613 TI - Surveillance of acute bacterial meningitis among children admitted to a district hospital in rural Mozambique. AB - BACKGROUND: Acute bacterial meningitis (ABM) remains an important cause of mortality among African children. Epidemiologic data with regard to ABM infection are necessary for prioritizing public health interventions. METHODS: We strengthened hospital-based surveillance of ABM among children admitted to Manhica District Hospital (Maputo, Mozambique). Cerebrospinal fluid (CSF) samples were collected from children admitted to the hospital who met clinical criteria of ABM. Laboratory determinations were performed. Clinical information and outcome of cases were recorded. RESULTS: During the first 12 months of surveillance, which began in January 2006, CSF samples were collected from 642 children <15 years of age with suspected meningitis (18% of all pediatric patients admitted to the hospital during that time). ABM was confirmed in 43 (7%) of the 642 cases. Haemophilus influenzae type b (Hib) (14 cases), pneumococcus (9 cases), and meningococcus (7 cases) represented approximately 70% of confirmed cases. Four of the 9 pneumococci were serotypes covered by the 7-valent pneumococcal conjugate vaccine. The case fatality rate among patients with ABM was 24% (8 of 33 with known outcome); an additional 8 patients left the hospital before discharge. The incidence of ABM was 85 per 100,000 population, which peaked at 2-12 months of age at 1078 cases per 100,000 population. All 9 pneumococci isolates were susceptible to chloramphenicol, and 8 were susceptible to penicillin (the additional 1 had intermediate resistance). For the 10 Hib isolates tested, only 1 was susceptible to chloramphenicol, and 5 were susceptible to ampicillin. CONCLUSION: These data reinforce the importance of ABM as a cause of hospital admission and death in rural sub-Saharan Africa. Most observed ABM cases could have been prevented by current pneumococcal and Hib conjugate vaccines. PMID- 19191614 TI - Incidence, seasonality, age distribution, and mortality of pneumococcal meningitis in Burkina Faso and Togo. AB - Streptococcus pneumoniae causes a substantial proportion of meningitis cases in the African meningitis belt; however, few reports exist to quantify its burden and characteristics. We conducted population-based and sentinel hospital surveillance of acute bacterial meningitis among persons of all ages in Burkina Faso and Togo in 2002-2006. S. pneumoniae and other organisms were identified by culture, polymerase chain reaction, or detection of antigen in cerebrospinal fluid (CSF). Information was collected on 2843 patients with suspected acute bacterial meningitis. CSF specimens were collected from 2689 (95%) of the patients; of these 2689, 463 (17%) had S. pneumoniae identified, 234 (9%) had Haemophilus influenzae type b identified, and 400 (15%) had Neisseria meningitidis identified. Of the 463 cases of S. pneumoniae meningitis, 99 (21%) were aged <1 year, 71 (15%) were aged 1-4 years, 95 (21%) were aged 5-14 years, and 189 (41%) were aged >or=15 years (age was unknown for 9 [2%]). In Burkina Faso, the annual incidence rate of pneumococcal meningitis was 14 cases per 100,000 persons, with annual incidence rates of 77, 33, 10, and 11 cases per 100,000 persons aged <1 year, <5 years, 5-14 years, and >or=15 years, respectively. The case-fatality ratio for S. pneumoniae meningitis was 47% (range for age groups, 44%-52%), and 53% of deaths occurred among those aged >5 years. S. pneumoniae meningitis had an epidemic pattern similar to that of N. meningitidis meningitis. Of 48 isolates tested for serotype, 18 were from children aged <5 years; of these 18, 3 isolates (17%) each were serotypes 1, 2, and 5, and 5 isolates (28%) were serotype 6A. The 7-, 10-, and 13-valent pneumococcal conjugate vaccines would cover 6%, 39%, and 67% of serotypes identified among children aged <5 years, respectively. Of the 30 serotypes identified for patients aged >or=5 years, 18 (60%) were serotype 1, whereas no other serotype constituted >10%. The 7-, 10-, and 13-valent vaccines would cover 7%, 70%, and 77% of serotypes. Epidemic pneumococcal meningitis in the African meningitis belt countries of Burkina Faso and Togo is common, affects all age groups, and is highly lethal. On the basis of a modest number of isolates from a limited area that includes only meningitis cases, 7-valent pneumococcal conjugate vaccine might have only a limited and short-term role. By contrast, the proposed 10- and 13-valent vaccines would cover most of the identified serotypes. To better inform vaccine policy, continued and expanded surveillance is essential to document serotypes associated with pneumonia, changes in serotype distribution across time, and the impact of vaccine after vaccine introduction. PMID- 19191616 TI - Case definitions for pneumococcal syndromes and other severe bacterial infections. PMID- 19191615 TI - Invasive pneumococcal disease in children aged <5 years admitted to 3 urban hospitals in Ibadan, Nigeria. AB - BACKGROUND: Streptococcus pneumoniae remains a major cause of childhood morbidity and mortality in the world. The introduction of pneumococcal conjugate vaccine in developing countries will be facilitated by a clearer understanding of the disease burden for bacterial causes of pneumonia and meningitis and the prevalent serotypes of S. pneumoniae. METHODS: We conducted a prospective, hospital-based surveillance for a 2-year period involving children aged 2-59 months at 3 urban hospitals in Ibadan, Nigeria, using standard microbiological methods with confirmation and further testing of isolates at the Medical Research Council Laboratories in The Gambia. RESULTS: There were 1210 cases overall: 481 (39.8%) were meningitis, 399 (33.0%) were pneumonia, and 330 (27.2%) were bacteremia clinical syndromes. There were 24 cases of definite meningitis, of which 9 were caused by S. pneumoniae, 11 by Haemophilus influenzae type b, and 4 by Klebsiella species. Of the 90 culture-positive pneumonia cases, 9 were caused by S. pneumoniae, 2 by H. influenzae type b, and 79 by other species. Among cases of bacteremia, the pathogen isolation rate was 28.8% (95 of 330); the isolated species included S. pneumoniae (3 isolates), Staphylococcus aureus (20 isolates), Klebsiella species (13 isolates), Salmonella species (15 isolates), and Escherichia coli (6 isolates). Of the 23 S. pneumoniae isolates, 11 were serotyped; the serotypes found were 5 (5 isolates), 19F (3 isolates), and 4 (3 isolates), and 1 isolate was nontypeable. These isolates were all susceptible to penicillin. Eight of 9 patients with definite pneumococcal meningitis died, whereas all patients with pneumococcal pneumonia and septicemia survived. CONCLUSIONS: Of the pneumococcal serotypes identified, 55% were covered by the licensed 7-valent pneumococcal conjugate vaccine, whereas all are covered by the 10- and 13-valent vaccines. PMID- 19191617 TI - Progress and future challenges in coordinated surveillance and detection of pneumococcal and Hib disease in developing countries. PMID- 19191618 TI - Standardizing surveillance of pneumococcal disease. AB - BACKGROUND: Surveillance for invasive pneumococcal disease has been conducted using a variety of case ascertainment methods and diagnostic tools. Interstudy differences in observed rates of invasive pneumococcal disease could reflect variations in surveillance methods or true epidemiological differences in disease incidence. To facilitate comparisons of surveillance data among countries, investigators of Pneumococcal Vaccines Accelerated Development and Introduction Plan-sponsored projects have developed standard case definitions and data reporting methods. METHODS: Investigators developed case definitions for meningitis, pneumonia, and very severe disease using existing World Health Organization guidelines and clinical definitions from Africa and Asia. Standardized case definitions were used to standardize reporting of aggregated results. Univariate analyses were conducted to compare results among countries and to identify factors contributing to detection of Streptococcus pneumoniae. RESULTS: Surveillance sites varied with regard to the age groups targeted, disease syndromes monitored, specimens collected, and laboratory methods employed. The proportion of specimens positive for pneumococcus was greater for cerebrospinal fluid specimens (1.2%-19.4%) than for blood specimens (0.1%-1.4%) in all countries (range, 1.3-38-fold greater). The distribution of disease syndromes and pneumonia severity captured by surveillance differed among countries. The proportion of disease cases with pneumococcus detected varied by syndrome (meningitis, 1.4%-10.8%; pneumonia, 0.2%-1.3%; other, 0.2%-1.2%) and illness severity (nonsevere pneumonia, 0%-2.7%; severe pneumonia, 0.2%-1.2%), although these variations were not consistent for all sites. Antigen testing and polymerase chain reaction increased the proportion of cerebrospinal fluid specimens with pneumococcus identified by 1.3-5.5-fold, compared with culture alone. CONCLUSIONS: Standardized case definitions and data reporting enhanced our understanding of pneumococcal epidemiology and enabled us to assess the contributions of specimen type, disease syndrome, pneumonia severity, and diagnostic tools to rate of pneumococcal detection. Broader standardization and more-detailed data reporting would further improve interpretation of surveillance results. PMID- 19191619 TI - Enhanced diagnosis of pneumococcal meningitis with use of the Binax NOW immunochromatographic test of Streptococcus pneumoniae antigen: a multisite study. AB - BACKGROUND: Accurate etiological diagnosis of meningitis in developing countries is needed, to improve clinical care and to optimize disease-prevention strategies. Cerebrospinal fluid (CSF) culture and latex agglutination testing are currently the standard diagnostic methods but lack sensitivity. METHODS: We prospectively assessed the utility of an immunochromatographic test (ICT) of pneumococcal antigen (NOW Streptococcus pneumoniae Antigen Test; Binax), compared with culture, in 5 countries that are conducting bacterial meningitis surveillance in Africa and Asia. Most CSF samples were collected from patients aged 1-59 months. RESULTS: A total of 1173 CSF samples from suspected meningitis cases were included. The ICT results were positive for 68 (99%) of the 69 culture confirmed pneumococcal meningitis cases and negative for 124 (99%) of 125 culture confirmed bacterial meningitis cases caused by other pathogens. By use of culture and latex agglutination testing alone, pneumococci were detected in samples from 7.4% of patients in Asia and 15.6% in Africa. The ICT increased pneumococcal detection, resulting in similar identification rates across sites, ranging from 16.2% in Nigeria to 20% in Bangladesh. ICT detection in specimens from culture negative cases varied according to region (8.5% in Africa vs. 18.8% in Asia; P< .001), prior antibiotic use (24.2% with prior antibiotic use vs. 12.2% without; P< .001), and WBC count (9.0% for WBC count of 10-99 cells/mL, 22.1% for 100-999 cells/mL, and 25.4% for >or=1000 cells/mL; P< .001 by test for trend). CONCLUSIONS: The ICT provided substantial benefit over the latex agglutination test and culture at Asian sites but not at African sites. With the addition of the ICT, the proportion of meningitis cases attributable to pneumococci was determined to be similar in Asia and Africa. These results suggest that previous studies have underestimated the proportion of pediatric bacterial meningitis cases caused by pneumococci. PMID- 19191620 TI - Surveillance of pneumococcal-associated disease among hospitalized children in Khanh Hoa Province, Vietnam. AB - BACKGROUND: To understand the epidemiology of childhood bacterial diseases, including invasive pneumococcal disease, prospective surveillance was conducted among hospitalized children in Nha Trang, Vietnam. METHODS: From April 2005 through August 2006, pediatricians at the Khanh Hoa General Hospital used standardized screening criteria to identify children aged <5 years who had signs and symptoms of invasive bacterial disease. All cerebrospinal fluid (CSF) and blood specimens collected were tested by bacterial culture. Selected culture negative specimens were tested for Streptococcus pneumoniae by antigen detection or for Haemophilus influenzae, Moraxella catarrhalis, Neisseria meningitidis, and S. pneumoniae by polymerase chain reaction (PCR). RESULTS: A total of 987 children were enrolled (794 with pneumonia, 76 with meningitis, and 117 with other syndromes consistent with invasive bacterial disease); 84% of children were aged 0-23 months, and 57% were male. Seven (0.71%) of 987 blood cultures and 4 (15%) of 26 CSF cultures were positive for any bacterial pathogen (including 6 for H. influenzae type b and 1 for S. pneumoniae). Pneumococcal antigen testing and PCR identified an additional 16 children with invasive pneumococcal disease (12 by antigen testing and 4 by PCR). Among children aged <5 years who lived in Nha Trang, the incidence rate of invasive pneumococcal disease was at least 48.7 cases per 100,000 children (95% confidence interval, 27.9-85.1 cases per 100,000 children). CONCLUSIONS: S. pneumoniae and H. influenzae type b were the most common causes of laboratory-confirmed invasive bacterial disease in children. PCR and antigen testing increased the sensitivity of detection and provided a more accurate estimate of the burden of invasive bacterial disease in Vietnam. PMID- 19191621 TI - Incidence of pneumococcal bacteremia requiring hospitalization in rural Thailand. AB - BACKGROUND: Population-based estimates of the incidence of invasive pneumococcal disease are unavailable for Thailand and other countries in Southeast Asia. We estimated the incidence of pneumococcal bacteremia cases requiring hospitalization in rural Thailand. METHODS: Blood cultures were performed on samples from hospitalized patients in 2 rural provinces where active, population based surveillance of community-acquired pneumonia is conducted. Blood cultures were performed at clinician discretion and were encouraged for all patients with suspected pneumonia and all children aged <5 years with suspected sepsis. Pneumococcal antigen testing was performed on positive blood culture specimens that failed to grow organisms on subculture. RESULTS: From May 2005 through June 2007, 23,853 blood culture specimens were collected overall, and 7319 were collected from children aged <5 years, which represented 66% and 47% of target patients, respectively. A total of 72 culture-confirmed pneumococcal bacteremia cases requiring hospitalization were identified. An additional 44 patients had media from positive blood cultures that yielded no growth on subculture but that had positive results of pneumococcal antigen testing. Of the 116 confirmed cases of bacteremia, 27 (23%) occurred in children aged <5 years; of these, 9 (33%) were confirmed by antigen testing only. The incidence of pneumococcal bacteremia cases requiring hospitalization among children aged <5 years had a range of 10.6 28.9 cases per 100,000 persons (incidence range if cases detected by antigen are excluded, 7.5-14.0 cases per 100,000 persons). CONCLUSIONS: Invasive pneumococcal disease is more common than was previously suspected in Thailand, even on the basis of estimates limited to hospitalized cases of bacteremia. These estimates, which are close to estimates of the incidence of hospitalized cases of pneumococcal bacteremia in the United States before introduction of pneumococcal conjugate vaccine, provide important data to guide public health care policy and to inform discussions about vaccine introduction in Thailand and the rest of Southeast Asia. PMID- 19191623 TI - Multihospital surveillance of pneumonia burden among children aged <5 years hospitalized for pneumonia in Bangladesh. AB - BACKGROUND: Pneumonia contributes substantially to childhood mortality in Bangladesh. We conducted a study to characterize the disease severity and risk factors for mortality among children hospitalized for pneumonia in Bangladesh. METHODS: We analyzed data on hospitalization, patient characteristics, and mortality collected by a multicenter hospital-based surveillance of pneumonia in Bangladesh. RESULTS: From May 2004 through April 2007, 4155 children aged 2-59 months who met a pneumonia case definition adopted by GAVI's Pneumococcal Vaccines Accelerated Development and Introduction Plan-sponsored surveillance networks were enrolled after blood culture specimens were obtained. The mean duration (+/-SD) from illness onset to hospital admission was 6+/- days; 1842 children (44%) received antimicrobial treatment before hospitalization, and an additional 924 (22%) received antimicrobial treatment after admission to the hospital. Bacteria were isolated from 161 (4%) of the 4155 specimens, including 10 (6%) Streptococcus pneumoniae isolates and 5 (3%) Haemophilus influenzae type b isolates. The case-fatality rate for pneumonia in the hospital was 4% (150 deaths), and the children who died did so after a median of 2 days of hospitalization (range, 0-24 days). Infancy was highly associated with death due to pneumonia (odds ratio [OR], 2.0; 95% confidence interval [CI], 1.3-3.2), as were very severe pneumonia (OR, 7.9; 95% CI, 5.6-11.2), a blood culture positive for bacteria (OR, 3.4; 95% CI, 2.0-5.8), severe malnutrition (OR, 4.6; 95% CI, 2.9-7.4), and delayed admission (mean [+/-SD] duration from illness onset to admission, 6+/-6 days, compared with 5+/-4 days for survivors; P< .04). CONCLUSIONS: The prevalence of pneumonia among children aged <5 years in hospitals in Bangladesh is high. However, the isolation rate of bacteria is low, possibly because of the high (68%) background use of antibiotics. Multiple risk factors associated with pneumonia case fatality suggest multiple strategies, including vaccines, to reduce pneumonia-related and overall child mortality in Bangladesh. PMID- 19191622 TI - Surveillance for invasive Streptococcus pneumoniae disease among hospitalized children in Bangladesh: antimicrobial susceptibility and serotype distribution. AB - BACKGROUND: Vaccines offer the prospect of primary disease prevention of pneumococcal disease in childhood. For introduction of such vaccines in developing countries, information about disease epidemiology is necessary. METHODS: We evaluated antimicrobial susceptibility and serotype distribution of invasive Streptococcus pneumoniae disease in children aged <5 years in a network of 7 hospitals in Bangladesh from May 2004 through May 2007. RESULTS: Of 17,969 blood cultures and 3765 cerebrospinal fluid cultures, 139 yielded S. pneumoniae isolates; 94 were from meningitis cases, 13 were from pneumonia cases, and 32 were from sepsis cases. Among the children with positive culture results, 73% were aged <12 months and 90% were aged <24 months. Complete resistance against penicillin, chloramphenicol, and cotrimoxazole was found in 0%, 6%, and 32% of isolates, respectively. Of the 37 serotypes observed, the predominant serotypes were 2 (17%), 1 (12%), 14 (7%), 5 (6%), 7F (6%), 45 (7%), and 12A (4%). Serotypes differed between meningitis cases and nonmeningitis cases, especially for serotype 2 (25% of meningitis cases vs. 0% of pneumonia cases; P < .001). The 7-, 10-, and 13-valent vaccines would cover 20% (95% confidence interval [CI], 13% 27%), 43% (95% CI, 35%-51%), and 50% (95% CI, 42%-58%) of these cases of invasive pneumococcal disease overall, with higher coverage of nonmeningitis cases, compared with meningitis cases (7-valent coverage, 23% vs. 18%; 10-valent coverage, 55% vs. 38%; 13-valent coverage, 66% vs. 42%). CONCLUSIONS: High levels of nonsusceptibility to cotrimoxazole and susceptibility to penicillin suggest that penicillin may be a drug of choice for treatment of invasive pneumococcal disease. Although serotype distribution is diverse, with changes over time and differences between syndromes observed, implementation of use of the currently available 10- or 13-valent vaccines would have a substantial impact on pneumococcal disease in Bangladesh. PMID- 19191624 TI - Neurodevelopmental sequelae in pneumococcal meningitis cases in Bangladesh: a comprehensive follow-up study. AB - BACKGROUND: Evaluation of the long-term impact of pneumococcal meningitis on surviving children and their families is critical to fully comprehending the burden of pneumococcal disease and to facilitating an evidence-based decision for the introduction of pneumococcal vaccine. This study was an investigation of the short- and long-term impacts of pneumococcal meningitis among Bangladeshi children. METHODS: Case patients with pneumococcal meningitis who were hospitalized between January 2006 and March 2007 were subjected to short-term follow-up within 30-40 days of discharge. Case patients discharged prior to January 2005 were selected for long-term follow-up at 6-24 months after the date of discharge. Both cohorts were enrolled from Dhaka Shishu Hospital, a pediatric hospital in Bangladesh. Healthy children matched for age, sex, socioeconomic status, and area of residence were recruited from the community as control subjects. During follow-up visits, case patients and control subjects were assessed for their physical and neurodevelopmental status by use of a standardized protocol. The impact of pneumococcal meningitis on siblings and the family as a whole was assessed by means of qualitative interviews. RESULTS: Neurodevelopmental assessments of the short-term follow-up cohort (n=51) revealed hearing, vision, mental, and psychomotor deficits in 33%, 8%, 41%, and 49% of the case patients, respectively. These deficits were 18%, 4%, 41%, and 35% in the long-term follow-up cohort (n=51), respectively. Such deficits were seen in only 2% of the control subjects, none of whom had vision or hearing deficits. CONCLUSIONS: In addition to the risk of death, pneumococcal meningitis in children causes severe disabilities among survivors, as well as disruption of the life of other siblings and family members. This study demonstrated that high rates of sequelae are associated with pneumococcal meningitis. Neurodevelopmental assessment during follow-up of patients with meningitis is critical to our understanding of the burden of the adverse consequences of pneumococcal disease. These data, along with the fact of poor access to health care, provide a compelling argument in favor of the introduction of pneumococcal vaccine, specifically in a setting where access to health care is poor and disabled children remain incapacitated because of a lack of resources and facilities. PMID- 19191625 TI - Use of multiple surveillance modalities to assess the epidemiology of Streptococcus pneumoniae infection in Bangladesh. AB - Measuring the broad impact of pneumococcal disease requires multiple surveillance modalities. Four major data sources elucidate the burden of pneumococcal disease in Bangladesh. The Bangladesh Demographic and Health Survey has identified pneumonia as the leading cause of childhood death. By extrapolation of mortality rates in the survey to the Bangladesh population, it has been estimated that approximately 90,000 children >1 month and <5 years of age die of pneumonia every year in Bangladesh. Through hospital-based surveillance, a wide range of pneumococcal serotypes leading to hospitalization and pneumonia have been identified as a leading cause of pediatric hospitalization. Urban community-based surveillance has demonstrated that invasive pneumococcal disease (IPD) is common in the community. Rural community-based surveillance has demonstrated that serious IPD is common in rural areas. Together, these data provide a strong scientific case for the importance of pneumococcal disease prevention to child health in Bangladesh and, therefore, the potential benefit of an effective vaccine. PMID- 19191628 TI - Childhood bacterial meningitis in Ulaanbaatar, Mongolia, 2002-2004. AB - BACKGROUND: Childhood bacterial meningitis is severe and largely preventable by vaccination. Few data on childhood bacterial meningitis in Northeast and Central Asia exist. Our aim was to determine the incidence and etiology of childhood bacterial meningitis in Ulaanbaatar, Mongolia. METHODS: We conducted prospective, population-based, active hospital surveillance for clinical meningitis in children 2 months to 5 years of age. Clinical data, blood, and cerebrospinal fluid were collected according to a standard protocol. Laboratory testing was performed at 2 reference laboratories in Ulaanbaatar. RESULTS: From February 2002 to January 2005, 201 suspected meningitis cases were identified in residents of Ulaanbaatar. The average annual incidence rate for confirmed and probable bacterial meningitis (cases with culture-negative, purulent cerebrospinal fluid) was 68 cases per 100,000 children aged 2 months to 5 years. The average annual incidence rate of confirmed cases was 28 cases per 100,000 children for Haemophilus influenzae type b meningitis, 11 cases per 100,000 children for pneumococcal meningitis, and 13 cases per 100,000 children for meningococcal meningitis. Adjusting for cases without complete cerebrospinal fluid information and culture-negative, probable bacterial cases, the estimated incidence rate was 40 cases per 100,000 children for H. influenzae type b meningitis, 15 cases per 100,000 children for pneumococcal meningitis, and 17 cases per 100,000 children for meningococcal meningitis. CONCLUSION: H. influenzae type b is the leading cause of childhood bacterial meningitis in Ulaanbaatar, and the incidence rate is higher than that reported from other Asian countries. These data supported the recent introduction of H. influenzae type b conjugate vaccine in Mongolia. Ongoing surveillance will monitor the impact of the vaccine. PMID- 19191629 TI - A multimodal training program for laparoscopic pyeloplasty. AB - PURPOSE: We introduced a multimodal training program for laparoscopic pyeloplasty (LP) and evaluated its safety, feasibility, and efficacy. METHODS: The program consisted of box-trainer training, animal-model training, and operative training. Five trainees with different experiences in open pyeloplasty and laparoscopy were exposed to the program. The mentor performed objective and subjective evaluations at each stage to ensure the training quality and operation safety. The perioperative parameters of five groups of patients who underwent LP by the trainees independently were evaluated. RESULTS: All trainees successfully finished the training program and independently performed five LPs under the supervision of the mentor. Five trainees spent different training times on the box-trainer and animal-model training,but acquired similar laparoscopic proficiency. There were no conversions to open procedures, transfusions, or deaths among the patients. No statistically significant difference was found in the operative time, estimated blood loss, postoperative hospital stay, and perioperative complications among the five trainees (P > 0.05). CONCLUSION: The multimodal training program can be used to train residents to perform advanced LP through step-by-step training from box trainer to animal model to clinical practice. The mentor-initiated approach is important to guarantee the training quality and safety. PMID- 19191631 TI - Editorial Comment. PMID- 19191630 TI - Two concomitant robot-assisted procedures in one anesthesia session: our experience. AB - PURPOSE: To evaluate the feasibility and safety of two concomitant robot-assisted procedures in the same patient in one anesthesia setting. PATIENTS AND METHODS: Data were reviewed for 15 patients for whom two concomitant robot-assisted surgeries at our center were planned. Preoperative details, port position, intraoperative problems encountered, surgical and anesthesia time, blood loss, complications, hospital stay, and postoperative recovery were analyzed. RESULTS: Fifteen patients underwent two concomitant robot-assisted procedures; at least one of the two surgeries was a urologic procedure. The second procedure added <50 mL of blood loss. Surgical time was increased by 58 minutes and 30 minutes in cases in which the primary procedure involved the upper and lower urinary tract, respectively, while anesthesia time was increased by 82 minutes and 30 minutes, respectively. There were no added complications during the second procedure, and hospital stay was not increased, being limited only by the first procedure. CONCLUSION: Planned robot-assisted surgery for two different operations is feasible in one anesthesia session, thereby reducing cost and overall hospital stay for the patient without any increased risk of perioperative morbidity. The decision to continue with the second procedure depends on the successful completion of the first procedure in a reasonable time and without any complications. PMID- 19191632 TI - Multidrug-resistant tuberculosis (TB) resistant to fluoroquinolones and streptomycin but susceptible to second-line injection therapy has a better prognosis than extensively drug-resistant TB. AB - Multidrug-resistant tuberculosis (TB) strains resistant to the fluoroquinolones and streptomycin but susceptible to second-line injection treatment would not be defined as extensively drug-resistant TB. In a cohort of 174 patients with multidrug-resistant TB, we demonstrated that 12 patients with multidrug-resistant TB strains resistant to the fluoroquinolones and streptomycin had significantly better initial and long-term outcomes, compared with 10 patients with extensively drug-resistant TB. PMID- 19191633 TI - Delayed microbial cure of lymphogranuloma venereum proctitis with doxycycline treatment. AB - Microbial cure of chlamydia proctitis (lymphogranuloma venereum [LGV] and non LGV) with doxycycline treatment was evaluated by chlamydia DNA and RNA persistence in anal swab specimens. In LGV proctitis, RNA persisted for up to 16 days. In non-LGV chlamydia proctitis, DNA was undetectable after 7 days. These findings support the Centers for Disease Control and Prevention's treatment recommendation of a 21-day doxycycline regimen for LGV proctitis and a 7-day regimen for non-LGV chlamydia proctitis. Delayed microbial cure of LGV proctitis should be considered in improved treatment regimens. PMID- 19191635 TI - Clinical practice guidelines for the management of candidiasis: 2009 update by the Infectious Diseases Society of America. AB - Guidelines for the management of patients with invasive candidiasis and mucosal candidiasis were prepared by an Expert Panel of the Infectious Diseases Society of America. These updated guidelines replace the previous guidelines published in the 15 January 2004 issue of Clinical Infectious Diseases and are intended for use by health care providers who care for patients who either have or are at risk of these infections. Since 2004, several new antifungal agents have become available, and several new studies have been published relating to the treatment of candidemia, other forms of invasive candidiasis, and mucosal disease, including oropharyngeal and esophageal candidiasis. There are also recent prospective data on the prevention of invasive candidiasis in high-risk neonates and adults and on the empiric treatment of suspected invasive candidiasis in adults. This new information is incorporated into this revised document. PMID- 19191636 TI - Duration and clearance of anal human papillomavirus (HPV) infection among women: the Hawaii HPV cohort study. AB - BACKGROUND: The association of anal cancer with human papillomavirus (HPV) infection is well established; however, little is known about the epidemiology of anal HPV in healthy women. We investigated patterns of duration and clearance of anal HPV infection in a cohort of healthy women in Hawaii. METHODS: Viral and nonviral determinants of anal HPV clearance were examined in a longitudinal cohort study of 431 sexually active women. At baseline and at 4-month intervals, interviews were conducted and cervical and anal cell specimens were obtained for detection of HPV DNA. RESULTS: Of the 431 women, 50% experienced a total of 414 incident anal HPV infections, reported at 1 clinic visits from baseline through a follow-up period of average duration of 1.2 years. Of these infections, 58% cleared during follow-up. The clearance rate for a high-risk anal infection was 9.2 per 100 woman-months (95% confidence interval [CI], 6.9-11.9 per 100 woman months), with a median duration of 150 days (95% CI, 132-243 days). The slowest clearing high-risk HPV types were HPV-59 (median clearance time, 350 days) and HPV-58 (median clearance time, 252 days). The median clearance times for HPV-16 and HPV-18, the predominant types associated with anal cancer, were 132 days and 212 days, respectively. Nonviral factors that delayed clearance of anal HPV included douching, long-term tobacco smoking, and anal sex. CONCLUSIONS: The majority of anal HPV infections resolve in a relatively short time. Although anal HPV is commonly acquired in healthy women, its rapid clearance suggests limited efficacy of HPV testing as an anal cancer screening tool. PMID- 19191637 TI - Duration of anal human papillomavirus infection among immunocompetent women: clues to anal cancer epidemiology and possible prevention strategies. PMID- 19191638 TI - Seroepidemiologic and occupational risk survey for Coxiella burnetii antibodies among US veterinarians. AB - BACKGROUND: Little is known about the occurrence of Q fever among veterinarians in the United States. In this study, we sought to estimate the prevalence of Coxiella burnetii antibodies among veterinarians and to identify risk factors for exposure. METHODS: We tested serum samples from 508 veterinarians who attended the 143rd American Veterinary Medical Association Annual Convention in 2006. Samples were screened using a Q fever IgG enzyme-linked immunosorbent assay (ELISA). Samples with positive or equivocal results of ELISA were confirmed using phase I and phase II IgG immunofluorescence antibody assays, and end point IgG titers were determined for samples with positive results. RESULTS: Antibodies against C. burnetii were detected in 113 (22.2%) of 508 veterinarians. Risk factors associated with seropositivity included age 46 years, routine contact with ponds, and treatment of cattle, swine, or wildlife. CONCLUSIONS: Veterinarians have a high level of exposure to C. burnetii, the causative organism of Q fever, especially those veterinarians who treat livestock. In this study, risk of C. burnetii seropositivity was also independently associated with contact with ponds. The role of exposure to standing bodies of water in infection is not usually considered and should be investigated in future studies. Additionally, the evidence of past infection with C. burnetii in >20% of veterinarians also highlights the need for use of appropriate personal protective equipment when treating animals that are potentially infected with C. burnetii. Physicians should consider the risk of infection with C. burnetii when treating ill veterinarians and others with potential occupational exposures. PMID- 19191639 TI - Reemergence of Q fever after 11 September 2001. PMID- 19191640 TI - Detection of circulating Chlamydophila pneumoniae in patients with coronary artery disease and healthy control subjects. AB - BACKGROUND: There is a long history of research suggesting that Chlamydophila pneumoniae is associated with coronary artery disease (CAD). C. pneumoniae in peripheral blood mononuclear cells (PBMCs) could serve as a risk factor for CAD if respiratory infection with C. pneumoniae spreads to atherosclerotic plaques through PBMCs or if infected plaques shed C. pneumoniae-laden PBMCs into the circulation. METHODS: PBMCs were collected from 86 case patients with abnormal coronary angiogram findings and from 91 age- and gender-matched healthy control subjects. The healthy control subjects were strictly defined as not having atherosclerosis on the basis of absence of both clinical atherosclerotic disease and traditional risk factors for CAD. PBMCs were probed for the presence of C. pneumoniae nucleic acid by 2 separate real-time polymerase chain reaction (PCR) assays that used primers for outer membrane protein A (ompA) and 16S ribosomal RNA. C. pneumoniae serologic findings were determined for both case patients and control subjects. RESULTS: Despite serologic findings indicating past exposure to C. pneumoniae (immunoglobulin G titer, >or=1:16) in 74% of case patients with CAD and control subjects, no C. pneumoniae DNA or RNA was detected in PBMCs from any of the case patients or control subjects, including a subset of 42 participants (18 with CAD) who had samples obtained serially over 8 months. Multiple laboratory controls, including controls for inhibition of PCR, produced expected results. CONCLUSIONS: The uniformly negative results with use of highly sensitive methods are in contrast to much of the published literature. Probing of PBMCs for the genes of C. pneumoniae does not appear useful as a noninvasive way of detecting the presence of C. pneumoniae in atheromatous lesions. PMID- 19191642 TI - Clostridium difficile infection: same incidence and worse prognosis? PMID- 19191641 TI - Health care-associated Clostridium difficile infection in adults admitted to acute care hospitals in Canada: a Canadian Nosocomial Infection Surveillance Program Study. AB - BACKGROUND: Clostridium difficile infection (CDI) is the most frequent cause of health care-associated infectious diarrhea in industrialized countries. The only previous report describing the incidence of health care-associated CDI (HA CDI) in Canada was conducted in 1997 by the Canadian Nosocomial Infection Surveillance Program. We re-examined the incidence of HA CDI with an emphasis on patient outcomes. METHODS: A prospective surveillance was conducted from 1 November 2004 through 30 April 2005. Basic demographic data were collected, including age, sex, type of patient ward where the patient was hospitalized on the day HA CDI was identified, and patient comorbidities. Data regarding severe outcome were collected 30 days after the diagnosis of HA CDI; severe outcome was defined as an admission to the intensive care unit because of complications of CDI, colectomy due to CDI, and/or death attributable to CDI. RESULTS: A total of 1430 adults with HA CDI were identified in 29 hospitals during the 6-month surveillance period. The overall incidence rate of HA CDI for adult patients admitted to these hospitals was 4.6 cases per 1000 patient admissions and 65 per 100,000 patient days. At 30 days after onset of HA CDI, 233 patients (16.3%) had died from all causes; 31 deaths (2.2%) were a direct result of CDI, and 51 deaths (3.6%) were indirectly related to CDI, for a total attributable mortality rate of 5.7%. CONCLUSIONS: The rates are remarkably similar to those found in our previous study; although we found wide variations in HA CDI among the participating hospitals. However, the attributable mortality increased almost 4-fold (5.7% vs. 1.5%; P<.001). PMID- 19191643 TI - Gram-negative bacteremia upon hospital admission: when should Pseudomonas aeruginosa be suspected? AB - BACKGROUND: Pseudomonas aeruginosa is an uncommon cause of community-acquired bacteremia among patients without severe immunodeficiency. Because tension exists between the need to limit unnecessary use of anti-pseudomonal agents and the need to avoid a delay in appropriate therapy, clinicians require better guidance regarding when to cover empirically for P. aeruginosa. We sought to determine the occurrence of and construct a model to predict P. aeruginosa bacteremia upon hospital admission. METHODS: A retrospective study was conducted in 4 tertiary care hospitals. Microbiology databases were searched to find all episodes of bacteremia caused by gram-negative rods (GNRs) 90 years, receipt of antimicrobial therapy within past 30 days, and presence of a central venous catheter or a urinary device. Among 250 patients without severe immunodeficiency, if no predictor variables existed, the likelihood of having P. aeruginosa bacteremia was 1:42. If >or= 2 predictors existed, the risk increased to nearly 1:3. CONCLUSIONS: P. aeruginosa bacteremia upon hospital admission in patients without severe immunodeficiency is rare. Among immunocompetent patients with suspected GNR bacteremia who have >or= 2 predictors, empirical anti-pseudomonal treatment is warranted. PMID- 19191644 TI - Severity of meningococcal disease associated with genomic bacterial load. AB - BACKGROUND: Diagnostic polymerase chain reaction (PCR) detection of Neisseria meningitidis has enabled accurate quantification of the bacterial load in patients with meningococcal disease. METHODS: Quantification of the N. meningitidis DNA level by real time-PCR was conducted on whole-blood samples obtained from patients presenting with meningococcal disease to hospitals throughout England and Wales over a 3-year period. Levels were correlated with clinical outcome, infecting serogroup, and host factors including, interleukin-1 genotype (IL-1). RESULTS: Bacterial loads were available for 1045 patients and were not associated with the age of the patient, delay in sample submission, or administration of antibiotics prior to admission. The median log bacterial load was higher in 95 patients who died (5.29 log(10)copies/mL; interquartile range, 4.41-6.30 log(10)copies/mL) than in 950 patients who survived (3.79 log(10)copies/mL; interquartile range, 2.87-4.71 log(10)copies/mL). Logistic regression revealed that age (odds ratio, 1.04 per 1-year increase in age) and bacterial load (odds ratio, 2.04 per log(10)-copies/mL increase) had a statistically significant effect on the risk of death. Infection with N. meningitidis serogroup C was associated with increased risk of death and an increased bacterial load. Also associated with a higher bacterial load were prolonged hospitalization (duration, >10 days); digit, limb, or soft-tissue loss; and requirement of hemodialysis. Carriage of IL-1RN(+2018) was associated with increased mortality (odds ratio, 2.14; P=.07) but not with a higher bacterial load. CONCLUSIONS: In meningococcal disease, bacterial load is associated with likelihood of death, development of permanent disease sequelae, and prolonged hospitalization. The bacterial load was relatively higher in patients infected with N. meningitidis serogroup C than in those infected with other serogroups. The effects of age and IL-1 genotype on mortality are independent of a high genomic bacterial load. PMID- 19191645 TI - Genetics in meningococcal disease: one step beyond. PMID- 19191646 TI - Antimotility agents for the treatment of Clostridium difficile diarrhea and colitis. AB - Antimotility agent use for the treatment of Clostridium difficile infection (CDI) is discouraged. We reviewed the literature and unpublished postmarketing surveillance reports regarding antimotility treatment of CDI. Twenty reports met inclusion criteria, describing 55 patients with CDI who were exposed to antimotility agents. All studies were case reports or series, with the exception of 1 retrospective review. Nineteen patients (35%) improved, with clinical resolution. Nine patients (16%) died, and 27 patients (49%) had unknown outcomes. Seventeen patients (31%) with CDI developed colonic dilation; 5 of these patients with severe CDI died. However, all patients who experienced complications or died were given antimotility agents alone initially, without an appropriate antibiotic. Twenty-three patients who received metronidazole or vancomycin coadministered with the antimotility agent experienced no complications. Evidence supporting the hypothesis that worsened outcomes are associated with antimotility therapy of CDI is lacking. Further study of the role of antimotility agents in providing symptomatic relief and reducing environmental contamination with infectious stool may be warranted. PMID- 19191647 TI - Antimotility agents for the treatment of Clostridium difficile infection: is the juice worth the squeeze? PMID- 19191648 TI - HIV-associated opportunistic infections--going, going, but not gone: the continued need for prevention and treatment guidelines. PMID- 19191649 TI - Extreme pyrexia and rapid death due to Staphylococcus aureus infection: analysis of 2 cases. AB - We describe unusual Staphylococcus aureus infections in 2 patients. The infections were characterized by extreme pyrexia and rapid death. Both causative organisms produced a deletion mutant form of toxic shock syndrome toxin-1 and variant enterotoxin C, which may have caused pyrexia and death. PMID- 19191650 TI - Streptococcus suis: an emerging human pathogen. AB - Streptococcus suis infection is acquired through exposure to contaminated pigs or pig meat. Over the past few years, the number of reported S. suis infections in humans has increased significantly, with most cases originating in Southeast Asia, where there is a high density of pigs. Increased awareness, improved diagnostics, and the occurrence of outbreaks have contributed to this increase. Meningitis and sepsis are the most common clinical manifestations of S. suis infection; hearing loss is a frequent complication. In this article, we provide an overview of the emergence and clinical manifestations of S. suis infection. PMID- 19191651 TI - Navigating the Web in search of resources on antimicrobial stewardship in health care institutions. AB - Bacterial resistance to antimicrobials has become a public health threat for which coordinated action at the international, national, and local level is needed. Current recommendations for the control of antimicrobial overuse and resistance in hospitals recommend various strategies, including antimicrobial stewardship programs. Several of these integrated and multidisciplinary antimicrobial management programs provide detailed information and recommendations on the Web. We performed a search of the most relevant and authoritative Web sites in English that were available without need for special registration or cost. The search excluded community-based programs, and we present only established programs or those providing expert information useful for building a hospital-based antimicrobial stewardship program. The overview of these Web sites may be useful either for institutions or individuals planning to implement such programs in their own health care institution or for educational purposes targeted at different professionals involved in improving antimicrobial practice. PMID- 19191652 TI - Lymphoma diagnosis and plasma Epstein-Barr virus load during vicriviroc therapy: results of the AIDS Clinical Trials Group A5211. AB - BACKGROUND: Lack of functional CCR5 increases the severity of certain viral infections, including West Nile virus and tickborne encephalitis. In a phase II trial of the investigational CCR5 antagonist vicriviroc (AIDS Clinical Trials Group protocol A5211), 4 lymphomas occurred in study patients who received vicriviroc. Because of the known association between unregulated Epstein-Barr virus (EBV) replication and lymphoma in immunocompromised patients, we evaluated whether vicriviroc exposure was associated with lymphoma EBV antigen positivity and/or had an effect on plasma levels of EBV DNA. METHODS: Clinical findings for all 4 patients enrolled in the A5211 study who developed lymphoma (2 Hodgkin and 2 non-Hodgkin) were reviewed, and tumor specimens were assessed for evidence of ongoing EBV replication. Longitudinal plasma samples from 116 patients in the A5211 study were analyzed, and EBV DNA was quantified by real-time polymerase chain reaction. RESULTS: Plasma EBV DNA was not detected in the 2 patients with non-Hodgkin lymphoma; both patients with Hodgkin lymphoma who had samples tested had EBV DNA levels <3200 copies/mL. One patient with Hodgkin lymphoma had a lymph node core biopsy specimen that was strongly positive for EBV; the other 3 lymphomas were histochemically EBV negative. None of the 116 patients with available samples experienced sustained increases in plasma EBV levels. CONCLUSIONS: CCR5 antagonism by vicriviroc treatment in treatment-experienced patients was not associated with reactivation of EBV infection. PMID- 19191653 TI - Characteristics and treatment outcomes among HIV-infected individuals in the Australian Trial in Acute Hepatitis C. AB - BACKGROUND: The Australian Trial in Acute Hepatitis C (ATAHC) is a National Institutes of Health-funded prospective cohort study of the natural history and efficacy of treatment in individuals with recently acquired hepatitis C. Enrollment is open to both human immunodeficiency virus (HIV)-infected and uninfected individuals. The aim of this article was to evaluate characteristics and virological outcomes among HIV-infected individuals enrolled in ATAHC. METHODS: Eligibility criteria included the first positive result of testing for anti-hepatitis C virus (HCV) antibody within 6 months and either clinical hepatitis diagnosed within the past 12 months or documented anti-HCV seroconversion within the past 24 months. RESULTS: Of the initial 103 patients enrolled, 27 (26%) were HIV infected. HIV-infected patients were more likely to be older, to have HCV genotype 1 infection and high levels of HCV RNA at baseline than were HCV-monoinfected patients. Sexual acquisition accounted for the majority (56%) of HCV infections among HIV-infected patients, compared with only 8% of HCV-monoinfected patients. The median duration from estimated HCV infection to treatment was 30 weeks. Treatment with 24 weeks of pegylated interferon and ribavirin resulted in rates of undetectability of HCV RNA of 95%, 90%, and 80% at weeks 12, 24, and 48, respectively. Undetectability at week 4 was achieved in 44% of patients and yielded positive and negative predictive values for sustained virological response of 100% and 33%, respectively. CONCLUSIONS: Significant differences were demonstrated between HIV-infected and HIV-uninfected individuals enrolled in ATAHC. Treatment responses among HIV-infected individuals with both acute and early chronic infection are encouraging and support regular HCV screening of high-risk individuals and early treatment for recently acquired HCV infection. PMID- 19191654 TI - Immunogenicity and safety of yellow fever vaccination for 102 HIV-infected patients. AB - BACKGROUND: Yellow fever vaccine (17DV) has been investigated incompletely in human immunodeficiency virus (HIV)-infected patients, and adequate immunogenicity and safety are of concern in this population. METHODS: In the Swiss HIV Cohort Study, we identified 102 patients who received 17DV while they were HIV infected. We analyzed neutralization titers (NTs) after 17DV administration using the plaque reduction neutralization test. NTs of 1:>or=10 were defined as reactive, and those of 1:<10 were defined as nonreactive, which was considered to be nonprotective. The results were compared with data for HIV-uninfected individuals. Serious adverse events were defined as hospitalization or death within 6 weeks after receipt of 17DV. RESULTS: At the time of 17DV administration, the median CD4 cell count was 537 cells/mm(3) (range, 11-1730 cells/mm(3)), and the HIV RNA level was undetectable in 41 of 102 HIV-infected patients. During the first year after vaccination, fewer HIV-infected patients (65 [83%] of 78; P = .01) than HIV-uninfected patients revealed reactive NTs, and their NTs were significantly lower (P < .001) than in HIV-uninfected individuals. Eleven patients with initially reactive NTs lost these reactive NTs or = 20 mmHg reduction from start of previous trial) and DBP response (< 90 or > or = 10 mmHg reduction from start of previous trial) increased at week 24 versus baseline from 51.8 to 69.3% and from 55.8 to 74.3%, respectively. At week 24, 110 (17.4%) patients achieved normal SBP/DBP (< 130/< 85 mmHg) and a further 24 (3.8%) patients achieved optimal SBP/DBP (< 120/< 80 mmHg). T80/H25 FDC was well tolerated; drug-related adverse events occurred in 29 (4.5%) patients. CONCLUSION: T80/H25 provides sustained and well-tolerated additive BP control, given alone or as part of a multi-antihypertensive therapy. PMID- 19191675 TI - Pharmacotherapy of patients with schizophrenia and substance abuse. AB - BACKGROUND: Comorbid substance-use disorder in schizophrenia patients is associated with poor clinical and functional outcome. OBJECTIVE: To provide evidence-based pharmacological treatment recommendations to improve the disease course of this patient group. METHODS: A comprehensive systematic review of the pharmacological studies in this subgroup of patients was performed and the available studies were discussed from the standpoint of evidence-based medicine. RESULTS/CONCLUSION: Slight advantages were found for second-generation antipsychotic agents over conventional antipsychotics with regard to improvement of distinct psychopathological symptoms, reduced craving, and greater reduction of substance use; these findings are mainly derived from open studies or case series. Additional treatment with antidepressants, depending on the patient's psychopathology, as well as with anti-craving agents should be considered. PMID- 19191676 TI - Pharmacotherapy of acute coronary syndrome: the ACUITY trial. AB - BACKGROUND: Bivalirudin, a direct thrombin inhibitor, has unique attributes including predictable pharmacokinetics, a reduction in bleeding complications and avoidance of heparin induced thrombocytopenia. OBJECTIVE: We critically review the role of bivalirudin in the management of patients with acute coronary syndrome in light of the Acute Catheterization and Urgent Intervention Triage Strategy (ACUITY) trial. METHODS: Data from the ACUITY trial was critically reviewed along with other contemporary data. RESULTS/CONCLUSION: In the ACUITY trial, bivalirudin monotherapy was associated with rates of ischemic events that were noninferior to those associated with heparin plus glycoprotein IIb/IIIa inhibition, but was associated with a significantly reduced incidence of major bleeding. Bivalirudin seems to be an attractive antithrombotic agent in patients with acute coronary syndrome undergoing early invasive management. PMID- 19191678 TI - Treatment options for AIDS patients with progressive multifocal leukoencephalopathy. AB - Progressive multifocal leukoencephalopathy (PML) is a demyelinating viral disease produced by the John Cunningham (JC) virus, which is ubiquitously distributed. Up to 80% of adults seroconvert to JC virus. Classically, PML is a life-threatening AIDS-defining disease of the CNS, usually occurring in severely immunocompromised individuals. Until now, and despite several therapeutic attempts, there is no specific treatment for PML. Soon after the widespread use of combination antiretroviral therapy (CART), several studies showed prolonged survival for patients with AIDS-associated PML who were treated with CART. The outcome of PML in patients receiving CART is unpredictable at disease onset. Prognostic markers are needed. The JC virus DNA detection in cerebrospinal fluid by nucleic acid amplification techniques and the CD4+ cell count are the most promising parameters. Higher levels of CD4+ cell counts were independently associated with an improved survival in different clinical observations. A summary of the main current knowledge about AIDS-related PML is presented. The most effective strategy is to optimize CART to completely suppress HIV-1 viral load and allow the best CD4+ T-cell immune recovery. Nowadays, AIDS-related PML is no longer an ultimately fatal disease. A substantial number of HIV-1-infected patients with this condition can improve with CART. PMID- 19191679 TI - A review of current anti-HCV treatment regimens and possible future strategies. AB - BACKGROUND: Hepatitis C virus (HCV) infection remains a serious health problem worldwide. The current standard treatment of HCV infection is pegylated interferon-alpha plus ribavirin, but this is clearly not sufficiently effective and tolerable. OBJECTIVE: To review current HCV treatment strategies and future options. METHODS: Review of major clinical trials or observational studies when no trial is available. RESULTS/CONCLUSION: Rates of sustained virologic response are widely variable, approximately 40-80%, depending on genotype, and even lower when HIV coinfection occurs. New agents, like small molecules that specifically target the HCV life cycle, may improve response rates; but safety is a concern. PMID- 19191680 TI - Pharmacotherapy of helminth infection. AB - BACKGROUND: In the first decade of the 21st century, worm infections are still very common, especially--but not exclusively--in the developing world. OBJECTIVE: To review the current pharmacotherapy of the major trematode, cestode and nematode infections of humans. METHODS: A systematic search of the Cochrane Databank of Controlled Trials and PubMed with MeSH terms (anthelmint(*) or treatment or therapy) and (cestoda or trematoda or nematoda or specific helminth species or specific medication). Further references were obtained from article biobliographies. RESULTS: Three hundred and twenty-six publications were selected for further review. CONCLUSION: Albendazole, praziquantel and ivermectin are the most important anthelmintics available, easy to use and active against most helminths. Diethylcarbamazine is used in loasis and lymphatic filariasis. Doxycycline can eliminate endosymbiotic bacteria of certain filariae, but its place in therapy needs to be further defined. In the treatment of cystic hydatid disease, a better, non-caustic protoscolicidal drug would diminish the complication rate of current puncture-aspiration-injection-reaspiration treatment. The reliance on so few drugs creates a dangerous situation for development of resistance. Triclabendazole is a welcome addition for fascioliasis. Tribendimidine, artemisinine derivatives and nitazoxanide are promising products, but their therapeutic place needs to be further defined. PMID- 19191677 TI - Tuberculosis pharmacotherapy: strategies to optimize patient care. AB - INTRODUCTION: The treatment of tuberculosis (TB) is a mature discipline, with more than 60 years of clinical experience accrued across the globe. The requisite Multi-drug treatment of drug-susceptible TB, however, lasts 6 months and has never been optimized according to current standards. Multi-drug resistant TB and TB in individuals coinfected with HIV present additional treatment challenges. OBJECTIVE: This article reviews the role that existing drugs and new compounds could have in shortening or improving treatment for TB. The key to treatment shortening seems to be sterilizing activity, or the ability of drugs to kill mycobacteria that persist after the initial days of multi-drug treatment. RESULTS: Among existing anti-TB drugs, the rifamycins hold the greatest potential for shortening treatment and improving outcomes, in both HIV-infected and HIV uninfected populations, without dramatic increases in toxicity. Clinical studies underway or being planned, are supported by in vitro , animal and human evidence of increased sterilizing activity--without significant increases in toxicity--at elevated daily doses. Fluoroquinolones also seem to have significant sterilizing activity. At present, at least two class members are being evaluated for treatment shortening with different combinations of first-line drugs. However, in light of apparent rapid selection for fluoroquinolone-resistant mutants, relative frequency of serious adverse events and a perceived need to 'reserve' fluoroquinolones for the treatment of drug-resistant TB, their exact role in TB treatment remains to be determined. Other possible improvements may come from inhaled delivery or split dosing (linezolid) of anti-TB drugs for which toxicity (ethionamide) or lack of absorption (aminoglycosides and polypeptides) precludes delivery of maximally effective, oral doses, once daily. New classes of drugs with novel mechanisms of action, nitroimidazopyrans and a diarylquinoline, among others, may soon provide opportunities for improving treatment of drug-resistant TB or shortening treatment of drug-susceptible TB. CONCLUSION: More potential options for improved TB treatment currently exist than at any other time in the last 30 years. The challenge in TB pharmacotherapy is to devise well-tolerated, efficacious, short-duration regimens that can be used successfully against drug resistant and drug-resistant TB in a heterogeneous population of patients. PMID- 19191681 TI - Pathophysiologic basis for the medical management of portal hypertension. AB - BACKGROUND: Portal hypertension is a potentially life-threatening complication of cirrhosis, resulting from increased intrahepatic resistance and portal inflow. OBJECTIVE: Given the complex nature of this disorder, a more complete understanding of the pathophysiology of portal hypertension is necessary to develop new therapies that target specific pathways that regulate portal pressure. METHODS: This review is based on a literature search of published articles and abstracts on the pathophysiology of portal hypertension, its complications and its treatment. RESULTS/CONCLUSION: A number of therapies have been developed or are under investigation for the treatment of portal hypertension and its complications. These agents may reduce mortality and improve quality of life for patients with advanced liver disease. PMID- 19191682 TI - Pharmacotherapy of gastroparesis. AB - Gastroparesis is a relatively common and often disabling condition that is characterized by a broad range of clinical presentation ranging from dyspeptic symptoms to nausea, vomiting, abdominal pain, malnutrition, frequent hospitalizations and incapacitation. The treatment of gastroparetic symptoms can be challenging to the gastroenterologist and the intensity of therapy varies with the physician's knowledge. Hence the determination that a patient is refractory to 'standard medical therapy' is an assessment that is subspeciality-based and could differ around the world depending on medications available. In this article, we review the use of available prokinetics, antiemetic agents, the approach for analgesia in the context of gastroparesis, and also discuss potential and evolving pharmacotherapies. The progress has been relatively limited as far as availability of new medications for gastroparesis is concerned; however, active research in developing newer prokinetics holds great promise for the future of management of this challenging entity. PMID- 19191683 TI - Clinical benefits versus shortcomings of diltiazem once-daily in the chronotherapy of cardiovascular diseases. AB - BACKGROUND: The introduction of chronotherapy (that is improving a drugs therapeutic efficacy by paralleling the drugs plasma levels to circadian rhythms) has recently become a focus of interest. OBJECTIVE: This article addresses the efficacy and potential shortcomings of chronotherapy, and focuses on one specific type of chronotherapy: a novel long-acting diltiazem formulation, DTZ-LA. METHODS: We reviewed the literature to assess the clinical benefits and shortcomings associated with DTZ-LA in the management of hypertension and angina. RESULTS/CONCLUSIONS: The clinical benefits of DTZ-LA outweigh its disadvantages when surrogate outcomes are evaluated, but it still remains to be determined whether chronotherapy benefits hard clinical outcomes. Nonetheless, chronotherapy has the potential to address the cardiovascular triggers that peak in the early morning hours when the preponderance of cardiovascular events occur, as well as providing better target organ protection compared with non-chronotherapeutic therapy. PMID- 19191684 TI - Somatostatin analog lanreotide in the treatment of castration-resistant prostate cancer (CRPC). AB - Prostate cancer is a common disease affecting males. Despite initial sensitivity to hormone treatment, prostate cancer eventually progresses to a castration resistant stage (CRPC), which carries an ominous prognosis. Lanreotide is a long acting somatostatin analog with the same properties with the native peptide. It has been shown to be highly efficacious in treating various hypersecretoty disorders and tumors. Lanreotide has been administered to patients with CRPC within a novel treatment concept, with the aim of targeting not only cancer cells but also various factors secreted in the tumor cell milieu that confer protection from apoptosis. Within this concept, lanreotide has been administered as part of the "antisurvival factor therapy" in combination with dexamethasone and a gonadotropin releasing hormone (GnRH) analog. It has also been given combined with oestrogens in patients with CRPC. The so far published series have documented a clinical response in many patients treated along with significant improvement in parameters related to quality of life. In view of these promising results, large-scale, randomized, controlled trials are warranted to clearly define the exact role of lanreotide and other somatostatin analogs in the treatment of patients with CRPC. PMID- 19191685 TI - Alogliptin: a new, highly selective dipeptidyl peptidase-4 inhibitor for the treatment of type 2 diabetes. AB - BACKGROUND: Alogliptin is a potent, highly selective dipeptidyl peptidase-4 inhibitor now undergoing clinical testing to support a new drug application for the treatment of type 2 diabetes. OBJECTIVE: To provide an overview of the mechanism by which alogliptin reduces hyperglycemia and to summarize the pharmacology, efficacy and safety of the drug. METHODS: A Medline search was conducted and abstracts and presentations from recent American Diabetes Association and European Association for the Study of Diabetes (EASD) meetings were reviewed. The results of Phase II and Phase III human studies were evaluated for clinical efficacy, safety and tolerability in patients with type 2 diabetes. RESULTS/CONCLUSIONS: Alogliptin is effective as a treatment for type 2 diabetes, either as monotherapy or in combination with metformin, thiazolidinediones, sulfonylureas and insulin. It is well tolerated and has an excellent safety profile. Additional studies are needed to evaluate the long-term safety and efficacy of alogliptin. PMID- 19191686 TI - New findings with old drugs for osteoporosis. AB - BACKGROUND: Postmenopausal osteoporosis is common and is associated with stooped posture, loss of height, back pain and fractures. OBJECTIVES/METHODS: This evaluation is of clinical outcome trials with tibolone (Long-Term Intervention of Fractures with Tibolone) and strontium ranelate (Spinal Osteoporosis Therapeutic Intervention) in postmenopausal osteoporosis. RESULTS: Although the Long-Term Intervention of Fractures with Tibolone trial established that tibolone decreased the incidence of vertebral and non-vertebral fractures in postmenopausal osteoporosis, it also showed that tibolone caused a small increase in the incidence of stoke. The Spinal Osteoporosis Therapeutic Intervention trial established that strontium ranelate decreased the incidence of vertebral fractures, but had little effect on the incidence of non-vertebral fractures. CONCLUSIONS: As some of the bisphosphonates (alendronate, risedronate, zoledronic acid) have been shown to prevent hip fractures without increasing the incidence of stroke, they should be preferred to tibolone and strontium in the treatment of postmenopausal osteoporosis. PMID- 19191688 TI - Synthesis of rare-metal absorbing polymer by three-component polyaddition through combination of chemo-selective nucleophilic and radical additions. AB - Sequential three-component polyaddition has been difficult, because the first addition typically consumes the two reactive monomers to produce a stable intermediate that cannot react smoothly with the remaining most stable monomer. The combination of nucleophilic and radical additions has overcome this thermodynamic problem. The first nucleophilic addition of diamines to cyclic dithiocarbonates produces bis(mercaptothiourethane)s that can attack diethynylbenzene radically, which consequently gives poly(thiourethane phenylenevinylenesulfide)s in high atom efficiency up to 95%. The resulting polymer extracted palladium and gold cations from their aqueous solutions in excellent efficiencies. PMID- 19191687 TI - Rate limiting step precedes C-C bond formation in the archetypical proline catalyzed intramolecular aldol reaction. AB - The archetypical proline-catalyzed intramolecular aldol reaction, the Hajos Parrish-Eder-Sauer-Wiechert reaction, has served as a model reaction for the mechanistic study of the ever-growing class of proline-catalyzed conversions. Experimental measurements of the (13)C kinetic isotope effects for this reaction show conclusively that carbon-carbon bond formation is not rate-limiting. PMID- 19191689 TI - Isomer restriction on a nanoparticle surface and enhanced blue emission. AB - 5-Mercapto-2,2'-bithiophene (BTSH) functionalized copper (Cu), silver (Ag), and gold (Au) nanoparticles (NPs) of various diameters have been synthesized. Emission with a peak around 455 nm is detected on irradiation with UV light. This emission, however, disappears upon photopolymerization of the NPs. The photophysical study of BTSH at room and low temperatures indicates that the trans isomer should be dominant when attached onto the surface of an NP. The electronic interaction among the ligands and NP core likely restricts the isomerization of BTSH and forms an emissive excited state. Therefore, we suggest that the trans isomer assembles on the NP surface giving the enhanced blue emission. As a control, no such isomer restriction in the case of 5-(5-mercaptopentyl)-2,2' bithiophene (BTC(5)SH) is observed when attached on the NP surface. PMID- 19191690 TI - Oxiranyl anion methodology using microflow systems. AB - Deprotonation of epoxides followed by trapping with electrophiles was carried out using microflow systems with varying temperature and residence time. Time dependence of the yields of products provides a deeper insight into chemical and configurational stabilities of oxiranyllithiums. With the thus-obtained information, reactions of oxiranyllithiums with various electrophiles were successfully carried out without decomposition and isomerization. PMID- 19191691 TI - Highly efficient incorporation of functional groups into aromatic main-chain polymer using iridium-catalyzed C-H activation and suzuki-miyaura reaction. AB - We report a mild, iridium-catalyzed borylation of aromatic polysulfone with bis(pinacolato)diboron [B(2)(pin)(2)] to form the corresponding borylated polysulfones up to high concentrations with nearly constant efficiency. The Suzuki-Miyaura cross-coupling reactions of the borylated polysulfones with functionalized aryl bromides allows installation of various functional groups such as ketone, amine, hydroxyl, and aldehyde to the polysulfone main chain in excellent conversion. PMID- 19191692 TI - Mechanism-based small molecule probes for labeling CD38 on live cells. AB - CD38 is a type II transmembrane glycoprotein with multiple functions. It acts as an ecto-enzyme as well as a receptor. The enzymatic activity catalyzes the formation of two potent Ca(2+) releasing agents: cyclic adenosine diphosphate ribose (cADPR) from nicotinamide adenine dinucleotide (NAD) and nicotinic acid adenine dinucleotide phosphate (NAADP) from NAD phosphate (NADP). The receptor function of CD38 leads to the phosphorylation of intracellular signaling proteins and the up-regulation of cytokine production in immune cells. These two functions of CD38 underlie its involvement in various biological processes, such as hormone secretion, immune cell differentiation, and immune responses. Clinically, CD38 is used as a negative prognosis marker for chronic lymphatic leukemia (CLL). However, a clear molecular understanding of CD38's role in physiology and pathology is still lacking. To facilitate the study of CD38 at cellular and molecular levels, here we report a mechanism-based method for fluorescently labeling CD38 on live cells. This labeling method does not interfere with the receptor function of CD38 and the downstream signaling. The labeling method is thus a useful tool to study the receptor function of CD38 in live cells. In addition, since the mechanism-based labeling also inhibits the enzymatic activity of CD38, it should be useful for dissecting the receptor function of CD38 without interference from its enzyme function in complicated biological processes. PMID- 19191693 TI - Silver-mediated fluorination of functionalized aryl stannanes. AB - We report a regiospecific silver-mediated fluorination of aryl stannanes. The presented reaction can afford complex fluoroarenes from readily available phenols in three steps. The operational simplicity and the broad substrate scope of the fluorination should render this reaction a useful tool for the synthesis of milligram to gram quantities of functionalized aryl fluorides. Silver-mediated oxidative transformations of aryl nucleophiles that proceed via bimetallic redox processes are a new avenue to develop carbon-heteroatom bond formations. PMID- 19191694 TI - Metal-free oxidative cross-coupling of unfunctionalized aromatic compounds. AB - A new strategy for mixed biaryl synthesis has been developed using the hypervalent iodine(III) reagents. The unique reactivities of the sigma-heteroaryl iodine(III) intermediates generated in situ are the key element for the unusual metal catalyst-free transformations and strict control of the product selectivities. PMID- 19191695 TI - Organization of nanoparticles in polymer brushes. AB - We have demonstrated a facile infiltration process, in which gold nanoparticles are assembled into block copolymer brushes. After solvent annealing, the polymer covered nanoparticles are either sequestered into the corresponding block copolymer domain or expulsed from the brush, depending on the shell density of the nanoparticles. PMID- 19191696 TI - Magnesium-catalyzed asymmetric direct aldol addition of ethyl diazoacetate to aromatic, aliphatic, and alpha,beta-unsaturated aldehydes. AB - Magnesium-catalyzed enantioselective aldol between ethyl diazoacetate and aromatic, aliphatic, and alpha,beta-unsaturated aldehydes affords alpha-diazo beta-hydroxy-esters in high enantioselectivities. Aldol adducts resulting from this asymmetric transformation are versatile intermediates toward the synthesis of several ester containing chiral building blocks. PMID- 19191697 TI - Polymer semiconductors for artificial photosynthesis: hydrogen evolution by mesoporous graphitic carbon nitride with visible light. AB - We investigated semiconductor characteristics for polymeric carbon nitride as a metal-free photocatalyst working with visible light and have shown that the efficiency of hydrogen production by photochemical water reduction can be improved by approximately 1 order of magnitude by introducing the right type of mesoporosity into polymeric C(3)N(4). We anticipate a wide rang of potential application of C(3)N(4) as energy transducers for artificial photosynthesis in general, especially with a 3D continuous nanoarchitecture. Moreover, the results of finding photoactivity for carbon nitride nanoparticles can enrich the discussion on prebiotic chemistry of the Earth, as HCN polymer clusters are unequivocal in the solar system. PMID- 19191699 TI - Explorations into neolignan biosynthesis: concise total syntheses of helicterin B, helisorin, and helisterculin A from a common intermediate. AB - Helicterins A and B (1 and 2), helisorin (3), and helisterculin A (4) are structurally unique natural products with the ability to combat the avian myeloblastosis virus. Biogenetically, their architectures are considered to be products of seemingly straightforward Diels-Alder, radical-based, or acid-induced dimerizations of common, simpler precursors. Yet, the pursuit of such blueprints in the laboratory has failed thus far in enabling their successful synthesis. Herein, we describe the first total syntheses of three of these natural products. Key features include the use of a building block distinct from Nature's likely starting material, highly complex retro Diels-Alder/Diels-Alder reaction cascades, an unconventional protecting group to achieve the proper balance of chemical reactivity on sensitive scaffolds, and several carefully developed reaction conditions that effectively balance competing reaction pathways. PMID- 19191698 TI - Dynamics of water and ions near DNA: comparison of simulation to time-resolved stokes-shift experiments. AB - Time-resolved Stokes-shift experiments measure the dynamics of biomolecules and of the perturbed solvent near them on subnanosecond time scales, but molecular dynamics simulations are needed to provide a clear interpretation of the results. Here we show that simulations using standard methods quantitatively reproduce the main features of TRSS experiments in DNA and provide a molecular assignment for the dynamics. The simulations reproduce the magnitude and unusual power-law dynamics of the Stokes shift seen in recent experiments [ Andreatta, D., et al. J. Am. Chem. Soc. 2005, 127, 7270 ]. A polarization model is introduced to eliminate cross-correlations between the different components contributing to the signal. Using this model, well-defined contributions of the DNA, water, and counterion to the experimental signal are extracted. Water is found to have the largest contribution and to be responsible for the power-law dynamics. The counterions have a smaller, but non-negligible, contribution with a time constant of 220 ps. The contribution to the signal of the DNA itself is minor and fits a 30 ps stretched exponential. Both time-averaged and dynamic distributions are calculated. They show a small subset of ions with a different coupling but no other evidence of substates or rate heterogeneity. PMID- 19191700 TI - Ruthenium-catalyzed N-alkylation of amines and sulfonamides using borrowing hydrogen methodology. AB - The alkylation of amines by alcohols has been achieved using 0.5 mol % [Ru(p cymene)Cl(2)](2) with the bidentate phosphines dppf or DPEphos as the catalyst. Primary amines have been converted into secondary amines, and secondary amines into tertiary amines, including the syntheses of Piribedil, Tripelennamine, and Chlorpheniramine. N-Heterocyclization reactions of primary amines are reported, as well as alkylation reactions of primary sulfonamides. Secondary alcohols require more forcing conditions than primary alcohols but are still effective alkylating agents in the presence of this catalyst. PMID- 19191701 TI - Green and efficient synthesis of sulfonamides catalyzed by nano-Ru/Fe(3)O(4). AB - The environmentally benign synthesis of carbon-nitrogen bonds continues to be an active and challenging field of chemical research. Here, a novel, environmentally benign method for the direct coupling of sulfonamides and alcohols is described. Despite the importance of sulfonamide derivatives as intermediates in drug synthesis, till now such transformations are rarely known. For the first time a domino dehydrogenation-condensation-hydrogenation sequence of alcohols and sulfonamides has been realized in the presence of a nanostructured catalyst. The magnetic property of the catalyst system allows for convenient isolation of the product and efficient recycling of the catalyst. A variety of coupling reactions of benzylic alcohols and sulfonamides including various heterocycles were successfully realized, often with >80% isolated yield. Advantageously, only one equivalent of the primary alcohol is consumed in the process. Mechanistic investigations of the competitive reactions of benzyl alcohol and d(7)-benzyl alcohol with p-toluenesulfonamide revealed a kinetic isotope effect (k(H)/k(D)) of 2.86 (+/-0.109) for the dehydrogenation of benzyl alcohol and 0.74 (+/-0.021) for the hydrogenation of N-benzylidene-p-toluenesulfonamide intermediate, which suggests dehydrogenation of the alcohol to be the rate determining step. PMID- 19191702 TI - Size-dependent kinetic enhancement in hydrogen absorption and desorption of the Li-Mg-N-H system. AB - High operating temperature and slow kinetics retard the practical applications of the Li-Mg-N-H system for hydrogen storage. To alleviate these problems, a first attempt was carried out by synthesizing Li(2)MgN(2)H(2) through sintering a mixture of Mg(NH(2))(2)-2LiNH(2) and investigating its size-dependent hydrogen storage performance. A dramatically enhanced kinetics for hydrogen absorption/desorption was achieved with a reduction in the particle size. For the dehydrogenation reaction, a three-dimensional diffusion-controlled kinetic mechanism was identified for the first time by analyzing isothermal hydrogen desorption curves with a linear plot method. The experimental improvement and mechanistic understanding on the dehydrogenation kinetics of the Li-Mg-N-H system shed light on how to further decrease the operating temperature and enhance the hydrogen absorption/desorption rate of the amide/hydride combined materials. PMID- 19191703 TI - Imperfect crystal and unusual semiconductor: boron, a frustrated element. AB - All elements, except for helium, appear to solidify into crystalline forms at zero temperature, and it is generally assumed that the introduction of lattice defects results in an increase in internal energy. beta-Rhombohedral boron, a thermodynamically stable form of elemental boron at high temperature, is known to have a large amount of partial occupied sites, seemingly in conflict with our common knowledge. By using lattice Monte Carlo techniques combined with ab initio calculations, we find that the beta-phase is stabilized by a macroscopic amount of intrinsic defects that are responsible not only for entropic effects but also for a reduction in internal energy. These defects enable the conversion of two center to three-center bonds and are accompanied by the presence of localized, nonconductive electronic states in the optical gap. In addition we find that the ab initio Ising model describing the partial occupancy of beta-boron has macroscopic residual entropy, suggesting that boron is a frustrated system analogous to ice and spin ice. PMID- 19191704 TI - Deprotection, tethering, and activation of a catalytically active metalloporphyrin to a chemically active metal surface: [SAc](4)P-Mn(III)Cl on Ag(100). AB - The adsorption and subsequent thermal chemistry of the acetyl-protected manganese porphyrin, [SAc](4)P-Mn(III)Cl on Ag(100) have been studied by high resolution XPS and temperature-programmed desorption. The deprotection event, leading to formation of the covalently bound thioporphyrin, has been characterized and the conditions necessary for removal of the axial chlorine ligand have been determined, thus establishing a methodology for creating tethered activated species that could serve as catalytic sites for delicate oxidation reactions. Surface-mediated acetyl deprotection occurs at 298 K, at which temperature porphyrin diffusion is limited. At temperatures above approximately 425 K porphyrin desorption, diffusion and deprotection occur and at >470 K the axial chlorine is removed. PMID- 19191705 TI - Gas-surface chemical reactions at high collision energies? AB - Most gas-surface chemical reactions occur via reaction of adsorbed species to form a thermal-energy ( approximately kT) product; however, some instances exist where an energetic projectile directly reacts with an adsorbate in a single collision event to form a hyperthermal product (with a kinetic energy of a few eV). Here we show for the first time that 30-300 eV F(+) bombardment of fluorinated Ag and Si surfaces produces "ultrafast" F(2)(-) products with exit energies of up to 90 eV via a multistep direct-reaction mechanism. Experiments conclusively show that the projectile F atom ends up in the fast molecular product despite the fact that the impact energy is far greater than typical bond energies. PMID- 19191706 TI - Coumarin-derived Cu(2+)-selective fluorescence sensor: synthesis, mechanisms, and applications in living cells. AB - A novel coumarin-based fluorogenic probe bearing the 2-picolyl unit (1) was developed as a fluorescent chemosensor with high selectivity and suitable affinity in biological systems toward Cu(2+) over other cations tested. The fluorescence on-off mechanism was studied by femtosecond time-resolved fluorescence (TRF) upconversion technique and ab initio calculations. The receptor can be applied to the monitoring of Cu(2+) ion in aqueous solution with a pH span 4-10. To confirm the suitability of 1 for biological applications, we also employed it for the fluorescence detection of the changes of intracellular Cu(2+) in cultured cells. The results indicate that 1 should be useful for the fluorescence microscopic imaging and the study on the biological functions of Cu(2+). PMID- 19191709 TI - Synthesis of deoxytetrahydrouridine. AB - The alpha-hydroxyamido functionality of 2'-deoxytetrahydrouridine (dTHU) makes this seemingly simple and generally useful compound difficult to obtain. Reported synthetic strategies produce extremely poor yields and multiple products, and full characterization data is not available. Described herein is a two-step approach for synthesizing dTHU in increased yields and purity; stability concerns are also addressed. Catalytic reduction (5% Rh/alumina) of 2'-deoxyuridine, followed by reduction with sodium borohydride as a limiting reagent, produces dTHU and limits formation of side products. Evidence was obtained for formation of a methoxy-substituted analogue during purification. By this strategy, dTHU of >95% purity can be obtained in 40% yield on a 150 mg scale. PMID- 19191707 TI - Comparative analysis of gene expression changes mediated by individual constituents of hemozoin. AB - Plasmodium protozoa, the source of malarial infections, catabolize large quantities of hemoglobin during an intraerythrocytic phase. During this process, free heme is detoxified through biomineralization into an insoluble heme aggregate, hemozoin (Hz). In its native state, Hz is associated with a variety of lipid peroxidation products including 4-hydroxy-2-nonenal (HNE). In the present study, gene expression profiles were used to compare responses to two of the individual components of Hz in a model macrophage cell line. LPS-stimulated RAW 264.7 cells were exposed to HNE and the synthetic form of Hz, beta-hematin (BH), for 6 or 24 h. Microarray analysis identified alterations in gene expression induced by exposure to HNE and opsonized BH (fold change, > or = 1.8; p value, < or = 0.01). Patterns of gene expression were compared to changes induced by an opsonized control latex bead challenge in LPS-stimulated cells and revealed that the BH response was predominantly phagocytic. Ingenuity Pathway Analysis demonstrated that HNE mediated a short-term oxidative stress response and had a prolonged effect on the expression of genes associated with categories of "Cell Cycle", "Cellular Assembly and Organization", "DNA Replication, Recombination, and Repair", and "Cellular Development". Comparisons of expression changes caused by BH and HNE with those observed during malarial infection suggest that BH and HNE are involved in inflammatory response modulation, altered NF-kappaB signal transduction, extracellular matrix (ECM) degradation, and dyserythropoiesis. HNE exposure led to several significant steady-state expression changes including repressed chemokine (C-C motif) ligand 5 (Ccl5), indicative of dyserythropoiesis, and a severe matrix metalloproteinase 9 (Mmp9)/tissue inhibitor of metalloproteinase 1 (Timp1) imbalance in favor of ECM proteolysis. PMID- 19191710 TI - Synthesis of fused cyclic systems containing medium-sized rings through tandem ROM-RCM of norbornene derivatives embedded in a carbohydrate template. AB - A general approach for the synthesis of fused cyclic systems containing medium sized rings (7-9) has been developed. The key steps involve a diastereoface selective Diels-Alder reaction of the dienophiles 4a-d attached to a furanosugar with cyclopentadiene and ring opening (ROM)-ring closing metathesis (RCM) of the resulting norbornene derivatives 10a-d and 11a-d. Diels-Alder reaction of the dienophiles 4a-d with cyclopentadiene in the absence of a catalyst produced 10a-d as the major product arising through addition of the diene to the unhindered Si face. The most interesting and new aspect of the Diels-Alder reaction of these dienophiles is the accessibility of the Re-face that was blocked by the alkenyl chains under Lewis acid catalysis producing the diastereoisomers 11a-d exclusively. The reversal of facial selectivity from an uncatalyzed reaction to a catalyzed one is unprecedented. The observed stereochemical dichotomy is attributed to rotation of the enone moiety along the sigma bond linking the sugar moiety during formation of the chelate 13. This makes the Re-face of the enone moiety in 4a-d unhindered. Diels-Alder reaction of the carbocyclic analogue 15 under Lewis acid catalysis produced a 1:1 mixture of the adducts 16 and 17 confirming the participation of sugar ring oxygen in chelate formation. Finally ROM-RCM of 10a-d and 11a-d with Grubbs' catalyst afforded the cis-syn-cis and cis anti-cis bicyclo-annulated sugars 21a-d and 23a-d, respectively, containing 7-9 membered rings. PMID- 19191711 TI - Lithium diisopropylamide-mediated ortholithiations: lithium chloride catalysis. AB - Ortholithiations of a range of arenes mediated by lithium diisopropylamide (LDA) in THF at -78 degrees C reveal substantial accelerations by as little as 0.5 mol % of LiCl (relative to LDA). Substrate dependencies suggest a specific range of reactivity within which the LiCl catalysis is optimal. Standard protocols with unpurified commercial samples of n-butyllithium to prepare LDA or commercially available LDA show marked batch-dependent rates--up to 100-fold--that could prove significant to the unwary practitioner. Other lithium salts elicit more modest accelerations. The mechanism is not discussed. PMID- 19191712 TI - Reductions of aliphatic and aromatic nitriles to primary amines with diisopropylaminoborane. AB - Diisopropylaminoborane [BH(2)N(iPr)(2)] in the presence of a catalytic amount of lithium borohydride (LiBH(4)) reduces a large variety of aliphatic and aromatic nitriles in excellent yields. BH(2)N(iPr)(2) can be prepared by two methods: first by reacting diisopropylamineborane [(iPr)(2)N:BH(3)] with 1.1 equiv of n butyllithium (n-BuLi) followed by methyl iodide (MeI), or reacting iPrN:BH(3) with 1 equiv of n-BuLi followed by trimethylsilyl chloride (TMSCl). BH(2)N(iPr)(2) prepared with MeI was found to reduce benzonitriles to the corresponding benzylamines at ambient temperatures, whereas diisopropylaminoborane prepared with TMSCl does not reduce nitriles unless a catalytic amount of a lithium ion source, such as LiBH(4) or lithium tetraphenylborate (LiBPh(4)), is added to the reaction. The reductions of benzonitriles with one or more electron-withdrawing groups on the aromatic ring generally occur much faster with higher yields. For example, 2,4 dichlorobenzonitrile was successfully reduced to 2,4-dichlorobenzylamine in 99% yield after 5 h at 25 degrees C. On the other hand, benzonitriles containing electron-donating groups on the aromatic ring require refluxing in tetrahydrofuran (THF) for complete reduction. For instance, 4-methoxybenzonitrile was successfully reduced to 4-methoxybenzylamine in 80% yield. Aliphatic nitriles can also be reduced by the BH(2)N(iPr)(2)/cat. LiBH(4) reducing system. Benzyl cyanide was reduced to phenethylamine in 83% yield. BH(2)N(iPr)(2) can also reduce nitriles in the presence of unconjugated alkenes and alkynes such as the reduction of 2-hexynenitrile to hex-5-yn-1-amine in 80% yield. Unfortunately, selective reduction of a nitrile in the presence of an aldehyde is not possible as aldehydes are reduced along with the nitrile. However, selective reduction of the nitrile group at 25 degrees C in the presence of an ester is possible as long as the nitrile group is activated by an electron-withdrawing substituent. It should be pointed out that lithium aminoborohydrides (LABs) do not reduce nitriles under ambient conditions and behave as bases with aliphatic nitriles as well as nitriles containing acidic alpha-protons. Consequently, both LABs and BH(2)N(iPr)(2) are complementary to each other and offer methods for the selective reductions of multifunctional compounds. PMID- 19191713 TI - A versatile synthesis of electroactive stilbenoprismands for effective binding of metal cations. AB - A versatile synthesis of a new class of polyaromatic receptors (stilbenoprismands) containing a Delta-shaped cavity similar to that of the pi prismand together with an intimately coupled electroactive stilbenoid moiety was accomplished via an efficient intramolecular McMurry coupling reaction. The presence of the Delta-shaped cavity in stilbenoprismands allows an efficient binding of a single silver cation as probed by (1)H NMR spectroscopy. Electron rich stilbenoprismands undergo a ready oxidation to their highly robust cation radical and dicationic salts. X-ray structure determination of a representative dicationic stilbenoprismand showed that the charges were largely localized on the tetraarylethylene moiety, which results in a twisting of the ethylenic C=C bond by approximately 35 degrees. Moreover, the electronic coupling among the stilbenoid and pi-prismand moieties in various stilbenoprismands was briefly probed by optical methods. PMID- 19191714 TI - Using ergosterol to mitigate the deleterious effects of ethanol on bilayer structure. AB - In wine fermentations, yeast is exposed to concentrated ethanol solutions. Ergosterol, a sterol that is found in lower eukaryotic membranes, helps preserve the structural integrity of yeast membranes in stressful environmental conditions. A premature arrest in ethanol production due to unknown metabolic changes in yeasts results in undesirably large concentrations of residual sugar and may be caused by the formation of an ethanol-induced interdigitated phase. We use atomistic molecular dynamics simulations to examine the induction of the interdigitated phase in model yeast membranes that contain either 0, 10, 20, 25 mol % ergosterol in ethanol concentrations of 0, 10, 15 vol %. The 25 mol % ergosterol system shows a similar level of interdigitation for the 0 and 10 vol % ethanol solutions, indicating that ergosterol molecules in this system are able to effectively counteract the disruptive behavior of ethanol molecules. However, at a 15 vol % ethanol solution, the amount of interdigitation triples and this ethanol concentration is similar to the concentrations found in stuck fermentations. The other three ergosterol concentrations studied (0, 10, 20 mol %) show larger quantities of interdigitation in the 10 vol % ethanol solution than the 0 vol % solution. Thus, the 25 mol % ergosterol bilayer, which is representative of the ergosterol concentrations seen in yeast membranes, is unique in the systems examined in its ability to delay the onset of ethanol induced interdigitation. The concentration of ergosterol affects the permeability of a fluid-phase bilayer, where the 10 mol % ergosterol bilayer is more permeable to ethanol than either a bilayer containing no ergosterol molecules or a bilayer containing 20/25 mol % ergosterol. This lipid permeability appears to be correlated with the existence of a lipid region whose lipids neither have direct contact with ergosterol molecules nor exhibit bulk lipid/lipid interactions. PMID- 19191715 TI - Light-induced change of configuration of the LHCII-bound xanthophyll (tentatively assigned to violaxanthin): a resonance Raman study. AB - Raman scattering spectra of light-harvesting complex LHCII isolated from spinach were recorded with an argon laser, tuned to excite the most red-absorbing LHCII bound xanthophylls (514.5 nm). The intensity of the nu(4) band (at ca. 950 cm-1) corresponding to the out-of-plane wagging modes of the C-H groups in the resonance Raman spectra of carotenoids appears to be inversely dependent on the probing laser power density. This observation can be interpreted in terms of excitation-induced change of configuration of the protein-bound xanthophyll owing to the fact that the intensity of this particular band is diagnostic of a chromophore twisting resulting from its binding to the protein environment. The comparison of the shape of the nu(4) band of a xanthophyll involved in the light induced spectral changes with the shape of the nu(4) band of the xanthophylls present in LHCII, reported in the literature, lets us conclude that, most probably, violaxanthin is a pigment that undergoes light-driven changes of molecular configuration but also the involvement of lutein may not be excluded. Possible physical mechanisms responsible for the configuration changes and physiological importance of the effect observed are discussed. PMID- 19191716 TI - In situ ESR/UV-vis-NIR and ATR-FTIR spectroelectrochemical studies on the p doping of copolymers of 3-methylthiophene and 3-hexylthiophene. AB - A combined spectroelectrochemical study by ESR/UV-vis-NIR as well as FTIR spectroscopy on the influence of the copolymer composition on the stabilization of charges upon electrochemical p-doping is presented. As compared to the parent homopolymers 3-hexylthiophene (3-HeTh) and 3-methylthiophene (3-MeTh) which seems to be irregular, FTIR studies of the copolymer of both monomers (copMeHeTh) point to a regioregular structure. The in situ ESR and UV-vis-NIR spectroelectrochemistry at higher doping levels of the polymeric materials proves bipolarons and polaron pairs as stable charged states in poly(3-hexylthiophene) as well as the copolymer copMeHeTh. During the p-doping of poly(3 methylthiophene) bipolarons are the dominating species at higher doping levels. It is demonstrated that only the simultaneous use of both the ESR and the UV-vis NIR spectroscopy enables the differentiation of polarons (paramagnetic) and polaron pairs (diamagnetic) in a conducting polymer. PMID- 19191717 TI - Relationship between morphological change and crystalline phase transitions of polyethylene-poly(ethylene oxide) diblock copolymers, revealed by the temperature dependent synchrotron WAXD/SAXS and infrared/Raman spectral measurements. AB - The phase transition behaviors of low-molecular-weight polyethylene-poly(ethylene oxide) (PE-b-PEO) diblock copolymers with the monomeric units of PE/PEO = 17/40 and 39/86 have been successfully investigated through the temperature-dependent measurements of wide-angle X-ray diffraction (WAXD), small-angle X-ray scattering (SAXS), infrared and Raman spectra, as well as thermal analysis. These diblock copolymers had been believed to show only order-to-disorder transition of lamellar morphology in a wide temperature region, but it has been found here for the first time that this copolymer clearly exhibits the three stages of transitions among lamella, gyroid, cylinder, and spherical phases in the heating and cooling processes. The WAXD and IR/Raman spectral measurements allowed us to relate these morphological changes to the microscopic changes in the aggregation states of PEO and PE segments. In the low-temperature region the PEO segments form the monoclinic crystal of (7/2) helical chain conformation and the PE segments of planar-zigzag form take the orthorhombic crystalline phase. These crystalline lamellae of PEO and PE segments are alternately stacked with the long period of 165 Angstroms. In a higher temperature region, where the PEO crystalline parts are on the way of melting but the PE parts are still in the orthorhombic phase, the gyroid morphology is detected in the SAXS data. By heating further, the gyroid morphology changes to the hexagonally packed cylindrical morphology, where the orthorhombic phase of PE segments is gradually disordered because of thermally activated molecular motion and finally transforms to the pseudohexagonal or rotator phase. Once the PE segments are perfectly melted, the higher-order structure changes from the cylinder to the spherical morphology. These morphological transitions might relate to the thermally activated motions of two short chain segments of the diblock copolymer, although the details of the transition mechanism are unclear at the present stage. PMID- 19191719 TI - New European guidelines for cardiovascular disease prevention in clinical practice. AB - In 2007, the Fourth Joint European Societies' Task Force on cardiovascular disease (CVD) prevention in clinical practice issued an update of the guidelines on how to prevent CVD. These guidelines are endorsed by 10 international societies interested in the subject. Particular attention is given to lifestyle changes, risk factor management, and the use of cardioprotective drugs. In this paper, the content of the guidelines is summarized with emphasis on what is new compared to the previous version and on what could be of particular interest to clinical chemists. For those who would like to read more on this, please refer to the full document (112 pages including 1105 references). PMID- 19191720 TI - The PCSK9 gene E670G polymorphism affects low-density lipoprotein cholesterol levels but is not a risk factor for coronary artery disease in ethnic Chinese in Taiwan. AB - BACKGROUND: An E670G polymorphism of the exon 12 of the proprotein convertase subtilisin/kexin type 9 (PCSK9) gene was recently found to be associated with increased plasma low-density lipoprotein cholesterol (LDL-C) levels and severity of coronary atherosclerosis. This case-control study tested for a possible link between this PCSK9 polymorphism and the risk of coronary artery disease (CAD) in an ethnic Chinese population in Taiwan. METHODS: The subjects included 202 CAD patients and 614 unrelated controls. Genotypes were determined via polymerase chain reaction, restriction mapping with MboII, and gel electrophoresis. RESULTS: Contradictory to the results of a previous report, a significantly lower level of LDL-C was noted in 670G carriers than in non-carriers (2.78+/-0.82 mmol/L vs. 3.02+/-0.85 mmol/L; p=0.029) among controls, after adjusting for age, gender, smoking, hypertension, diabetes mellitus, body mass index, and use of lipid lowering agents. The 670G carrier was identified less frequently in patients with CAD than in controls (9.9% vs. 11.9%), but the difference was not significant in a multivariable logistic regression analysis (odds ratio=0.73; 95% CI=0.24-2.22; p=0.575). The G allele also occurred at similar frequencies in the two groups (5.0% vs. 6.0%; p=0.421). CONCLUSIONS: These results indicate that the E670G polymorphism of the PCSK9 gene modulates plasma LDL-C levels, but that it is not a risk variant for CAD in ethnic Chinese in Taiwan. PMID- 19191721 TI - Polymorphisms and plasma soluble levels of E-selectin in patients with chronic hepatitis B virus infection. AB - BACKGROUND: Infections with hepatitis B virus (HBV) may lead to a distinct clinical outcome which is partially related to host genetic variability. Our aim was to investigate the relationships between the polymorphisms of the E-selectin gene and disease progression in a HBV-infected Chinese Han population, and also to determine the plasma soluble E-selectin (sE-selectin) levels in this population. METHODS: Genomic DNA was extracted from the peripheral blood of 367 HBV carriers and 281 healthy controls. Two polymorphisms (PstI for A561C and HphI for G98T) of the E-selectin gene were analyzed by polymerase chain reaction restriction fragment length polymorphism. Circulating sE-selectin levels were measured by specific enzyme-linked immunosorbent assay (ELISA). RESULTS: The frequency of the C allele (AC or CC) of the A561C polymorphism was significantly higher in patients with liver cirrhosis (LC) compared to controls (p=0.002). There was no difference in allele distribution of the G98T polymorphism. But in patients with LC, classified according to the Child-Pugh classification, the frequency of the T carrier (GT and TT) was significantly different between Child Pugh class A and class B plus C (p=0.009). Levels of plasma soluble E-selectin (sE-selectin) were significantly increased in HBV carriers with chronic hepatitis (CH) and LC (mean+/-SD 68.94+/-34.09 and 43.39+/-18.00 ng/mL) compared to controls (13.96+/-7.50 ng/mL) (p<0.01). In the LC subgroup, levels of sE-selectin were significantly decreased from Child-Pugh class A to class C (p<0.05). In each group, individuals with the C allele showed higher sE-selectin levels than those with the A allele (p<0.05). CONCLUSIONS: This is the first report describing the association between E-selectin polymorphisms and HBV-related chronic liver diseases. Our data suggest that the A561C polymorphism of the E-selectin gene may be associated with disease progression in patients with chronic HBV infection and control the expression of plasma soluble levels, while the G98T polymorphism may be related to fibrotic severity in the Chinese population. PMID- 19191722 TI - Effect of high levels of human chorionic gonadotropin and estradiol on the severity of hyperemesis gravidarum. AB - BACKGROUND: Serum human chorionic gonadotropin (hCG) and estradiol levels are higher in women with hyperemesis gravidarum (HG) than in unaffected pregnant controls. We postulated that higher levels of these hormones may be associated with clinically more severe HG. The aim of this study was to evaluate the effect of maternal hCG and estradiol levels on the severity of HG. METHODS: A prospective study was performed on 167 first trimester women hospitalized for HG. Venous blood was taken for hCG and estradiol levels. Scattergrams were plotted for hCG or estradiol levels vs. gestational age. A curve of best fit was drawn. Women were categorized into two groups according to their position above or below the curve. Prolonged hospital stay (> or =4 days) was used as a marker for HG severity. Multivariable logistic regression analysis was used to control for differences in characteristics, laboratory results on admission, and treatment received. RESULTS: After adjustment, high hCG level adjusted odds ratio (AOR) (2.2, 95% CI 1.0-4.9, p=0.04) and hyponatremia AOR (2.8, 95% CI 1.2-6.6, p=0.02) were independently associated with prolonged hospital stay. With bivariate analysis, high estradiol level was not associated with prolonged stay. CONCLUSIONS: High hCG but not estradiol is associated with more severe HG. PMID- 19191724 TI - High concentrations of haptocorrin interfere with routine measurement of cobalamins in human serum and milk. A problem and its solution. AB - BACKGROUND: Human milk and occasional serum samples contain high concentrations of unsaturated haptocorrin, which influence accurate measurement of cobalamins. METHODS: Cobalamins in serum samples spiked with increasing amounts of unsaturated haptocorrin were measured employing the Centaur, Cobas and Architect analysers. Cobinamide-coated EAH sepharose was employed for pretreatment of the samples. Human milk samples were collected from 24 healthy mothers. Haptocorrin was measured by ELISA. RESULTS: The measured concentration of cobalamins either increased (Centaur analyser) or decreased (Architect, Cobas analysers) significantly for haptocorrin >10 nM, and was 220%, 52% or 45% of the expected values in a serum sample containing 50 nM haptocorrin. Following pretreatment with cobinamide-sepharose, the expected cobalamin concentration was obtained (Centaur). The milk samples contained 4.5-180 nM haptocorrin. In samples containing >10 nM haptocorrin (n=19), the median concentration of cobalamins decreased from 1.3 nM to 0.67 nM after pretreatment with cobinamide-sepharose. CONCLUSIONS: Haptocorrin in concentrations above 10 nM influences measurement of cobalamins giving rise to falsely elevated or decreased results. Removal of unsaturated haptocorrin by pretreatment with cobinamide-sepharose solves the problem. PMID- 19191723 TI - Methodology for monitoring cytomegalovirus infection after renal transplantation. AB - BACKGROUND: The aim of this study was to evaluate the diagnostic value of different detection methods for cytomegalovirus (CMV) infection after renal transplantation and also to establish a system to monitor therapy for CMV infection. METHODS: We retrospectively studied 1516 renal transplant recipients from June 1994 to December 2006. All patients were screened for CMV-DNA. A total of 1402 patients had received CMV-IgG/IgM detection since June 1996 and 660 had received CMV antigen detection since June 2000. RESULTS: A total of 664 (43.8%) recipients developed CMV infection. The sensitivity, specificity and Youden index of the three methods, respectively, were 18.84%, 100% and 0.1884 for ELISA, 91.86%, 82.98% and 0.7484 for PCR, and 88.06%, 96.95% and 0.8501 for the CMV-pp65 antigenemia test. The sensitivity and specificity of the two combined detection methods (CMV-DNA and CMV-pp65) for post-operation CMV infection were 93.49% and 99.06%; the two detection methods had significant dependability (p<0.05) in diagnosis of CMV infection and in evaluation of therapeutic effect of antiviral drugs. CONCLUSIONS: Only ELISA can be used as a screening index in order to distinguish whether the donors or recipients are infected with CMV or not. CMV pp65 antigenemia can help guide clinical therapy for CMV infection. CMV-pp65 and CMV-PCR combined together provide a more effective method to monitor CMV infection and predict its outcome. PMID- 19191725 TI - Inter- and intra-individual variations of urinary endogenous metabolites in healthy male college students using (1)H NMR spectroscopy. AB - BACKGROUND: Most human metabolomics studies have shown that spectral outputs of (1)H nuclear magnetic resonance fingerprinting are strongly influenced by inter- and intra-individual variations; however, few studies have been performed to evaluate the inter- and intra-individual variations in urinary endogenous metabolites. METHODS: We recruited 30 male college students to evaluate the factors affecting intra- and inter-individual variations in urinary endogenous metabolites. Statistical analysis for variations in urinary metabolites was performed after eliminating outliers found in principal component analysis (PCA) plots. RESULTS: Inter-individual variations were relatively low for 2 oxoglutarate, succinate, citrate, dimethylglycine, and taurine, but high for trimethylaminoxide (TMAO), hippurate, and lactate. Intra-individual variations for 2-oxoglutarate, citrate, dimethylglycine, and taurine were relatively low, but high for TMAO and hippurate. The factors affecting inter-individual variation of lactate were age, body mass index, beverages, and alcohol, whereas the factors affecting intra-individual variation of lactate were age and fish. Kim Chi intake affected the inter-individual variation of succinate, citrate, TMAO, and hippurate; however, it did not affect the intra-individual variation of endogenous metabolites. CONCLUSIONS: Our results showed that inter- and intra individual variations in urinary endogenous metabolites were very large, and significant factors affecting inter- and intra-individual variation were diverse, even after eliminating outliers in PCA analysis. PMID- 19191726 TI - Preanalytical errors in primary healthcare: a questionnaire study of information search procedures, test request management and test tube labelling. AB - BACKGROUND: Most errors in laboratory medicine occur in the preanalytical phase and are the result of human mistakes. This study investigated information search procedures, test request management and test tube labelling in primary healthcare compared to the same procedures amongst clinical laboratory staff. METHODS: A questionnaire was completed by 317 venous blood sampling staff in 70 primary healthcare centres and in two clinical laboratories (response rate = 94%). RESULTS: Correct procedures were not always followed. Only 60% of the primary healthcare staff reported that they always sought information in the updated, online laboratory manual. Only 12% reported that they always labelled the test tubes prior to drawing blood samples. No major differences between primary healthcare centres and clinical laboratories were found, except for test tube labelling, whereby the laboratory staff reported better practices. Re-education and access to documented routines were not clearly associated with better practices. CONCLUSIONS: The preanalytical procedure in the surveyed primary healthcare centres was associated with a risk of errors which could affect patient safety. To improve patient safety in laboratory testing, all healthcare providers should survey their preanalytical procedures and improve the total testing process with a systems perspective. PMID- 19191727 TI - Age and hormone replacement therapy as factors influencing androgen levels in the postmenopausal female. AB - BACKGROUND: The changes in androgen levels after the menopause and the effects of hormone replacement therapy (HRT) itself and the mode of HRT have not been established. The objectives of this study were to document the effect of age on androgen levels in a normal population of postmenopausal women and to investigate the effect of oral or transdermal HRT on androgen, oestradiol and sex hormone binding globulin (SHBG) values. METHODS: A cross-sectional study was conducted on 182 postmenopausal females aged 45-100 years, randomly recruited from the community. Serum levels of dehydroepiandrosterone sulphate (DHEAS), androstenedione, testosterone, free androgen index (FAI) and SHBG in a reference group of women, subdivided by age, menopausal status, HRT replacement and mode of HRT replacement. RESULTS: Age was a significant factor affecting androstenedione (<55, > or =55 years) and DHEAS (<65, > or =65 years). HRT status was found to influence values for androstenedione, SHBG and FAI. Neither HRT nor age significantly influenced testosterone. CONCLUSIONS: The observations suggest the need to provide age-specific reference intervals for androstenedione and DHEAS but not for testosterone and SHBG. The significant effect of HRT treatment would also suggest a need for treatment associated reference intervals for androstenedione, SHBG and FAI. Suggested reference intervals for each of these partitioned groups are presented. PMID- 19191728 TI - Preheparin serum lipoprotein lipase mass interacts with gender, gene polymorphism and, positively, with smoking. AB - BACKGROUND: Correlates of preheparin serum lipoprotein lipase (LPL) mass and its associations with the likelihood of metabolic syndrome (MS) and coronary heart disease (CHD) were investigated. METHODS: A cross-sectional study was carried out in a population sample (n=352, median age 55). MS was defined according to modified Adult Treatment Panel III criteria. RESULTS: Age-adjusted geometric mean preheparin LPL concentrations were 58.6+/-1.04 ng/mL in men and 66.9+/-1.03 ng/mL in women (p<0.004). A positive interaction with both the LPL X447 allele (p<0.034) and age-adjusted smoking status (p=0.026 in men and p=0.11 women) was observed. LPL mass was significantly correlated in both genders with high-density lipoprotein (HDL)-cholesterol and inversely with triacylglycerol levels and HOMA index. In multiple linear regression analysis, LPL mass was significantly associated with genotype, gender, age, adiponectin, smoking status and HDL cholesterol, and in women with C-reactive protein after adjustment for body mass index, triacylglycerol and insulin. Significantly low sex- and age-adjusted serum LPL mass was observed in cases of MS, hypertension and CHD. Logistic regression analysis after adjustment for age, sex, adiponectin and S447X polymorphism demonstrated that LPL mass was inversely associated with CHD in men and both genders (p=0.02), with hypertension confined to women (p=0.04) and with MS likelihood in both genders combined and women [odds ratio 1.51 (95% CI 1.14-2.00) for halving the likelihood]. CONCLUSIONS: LPL X447 genotype, female gender and smoking habit interact in increasing preheparin serum LPL mass in Turkish adults. Serum LPL mass is inversely associated with MS and CHD, independent of confounders, and probably reflects insulin sensitivity. PMID- 19191729 TI - Comparison of calcium, magnesium, sodium, potassium, zinc, and creatinine concentration in 24-h and spot urine samples in women. AB - BACKGROUND: The 24-h urine sample is considered as the most reliable material for testing many but not necessarily all constituents in urine. However, its collection is tedious for both patients and research participants. The aim of this study was to compare concentrations of essential elements calcium (Ca), magnesium (Mg), sodium (Na), potassium (K), and zinc (Zn) in 24-h and spot urine samples. METHODS: Urine samples were collected from 143 generally healthy women, aged 30-79 years. Fasting spot urine was collected immediately after the end of the 24-h collection, therefore being of the same content as the first morning urine which ended the 24-h collection. Elements were analyzed by flame atomic absorption/emission spectrometry and expressed as mg/g and/or mmol/mol of creatinine (Cr). Spearman rank order correlations between 24-h and spot urine were carried out for each element. Ratios of elements in 24-h to spot urine samples were calculated to estimate the element-proportion of spot in the 24-h sample. RESULTS: All coefficients of correlation between 24-h and spot urine of measured elements and Cr were significant (p<0.05): Zn (0.637), Mg (0.623), Ca (0.603), Na (0.452), K (0.396), and Cr (0.217). Ratios of 24-h to spot urine samples for each element (except K) were similar and close to 2, indicating uniform proportion of elements from spot urine sample in the 24-h sample. In addition, a high correlation between various pairs of elements was obtained in both 24-h and spot urine; the highest being between Na/Ca (0.435) and (0.578), respectively. This is in accordance with theoretical presumptions and previous findings regarding those relationships. CONCLUSIONS: Although replacing burdensome 24-h urine collection with spot urine sampling might not provide the solution in all cases, our results show that for the elements analyzed, spot urine could be a reliable alternative. PMID- 19191730 TI - Performance of free beta-human chorionic gonadotrophin (free beta-hCG) and pregnancy associated plasma protein-A (PAPP-A) analysis between Delfia Xpress and AutoDelfia systems in The Netherlands. AB - BACKGROUND: The VU University Medical Center (VUmc) was the first hospital in the Netherlands to introduce the Delfia Xpress for the analysis of free beta-human chorionic gonadotrophin (beta-hCG) and pregnancy associated plasma protein-A (PAPP-A) in the first trimester screening program for Down syndrome. Since then, others have implemented this system. In this study, we tested the equality of measurements for free beta-hCG and PAPP-A between Delfia Xpress systems and one AutoDelfia system. METHODS: A total of 40 serum samples were aliquoted and stored at -20 degrees C. Samples were analyzed by six Delfia Xpress systems and one AutoDelfia system over a time period of 2 years. RESULTS: The relationships between free beta-hCG and PAPP-A were excellent for the different Delfia Xpress systems (r>0.99, p<0.0001). For PAPP-A, the agreement between the main system at VUmc and five other systems was linear with slopes between 0.99 and 1.06. Similarly, agreement for free beta-hCG was linear with slopes between 0.99 and 1.09. Likewise, agreement for PAPP-A and free beta-hCG was excellent for the AutoDelfia vs. the main Delfia Xpress at the VUmc (r>0.99, p<0.0001). For both PAPP-A and free beta-hCG, the relationships were linear with slopes of 1.08 and 1.07. CONCLUSIONS: We demonstrate an excellent agreement for the analysis of PAPP A and free beta-hCG between Delfia Xpress systems and one AutoDelfia system. PMID- 19191731 TI - Interference of IgM-lambda paraprotein with biuret-type assay for total serum protein quantification. PMID- 19191732 TI - Evaluation of BD Vacutainer PST II tubes for a wide range of immunoassays. PMID- 19191733 TI - Native prothrombin as an alternative to prothrombin time determination for monitoring oral anticoagulant therapy. PMID- 19191739 TI - AsH3 ultraviolet photochemistry. AB - High-n Rydberg time-of-flight spectroscopy has been used to study the 193.3 nm photolysis of AsH(3). The center-of-mass translational energy distribution for the 1-photon process, AsH(3) + h nu --> AsH(2) + H, P(E(c.m.)), indicates that AsH(2) internal excitation accounts for approximately 64% of the available energy [i.e., h nu - D(0)(H(2)As - H)]. Secondary AsH(2) photodissociation also takes place. Analyses of superimposed structure atop the broad P(E(c.m.)) distribution suggest that AsH(2) is formed with significant a-axis rotation as well as bending excitation. Comparison of the results obtained with AsH(3) versus those of the lighter group-V hydrides (NH(3), PH(3)) lends support to the proposed mechanisms. Of the group-V hydrides, AsH(3) lies intermediate between the nonrelativistic and relativistic regimes, requiring high-level electronic structure theory. PMID- 19191736 TI - Structural and functional studies of QdtC: an N-acetyltransferase required for the biosynthesis of dTDP-3-acetamido-3,6-dideoxy-alpha-D-glucose. AB - 3-Acetamido-3,6-dideoxy-alpha-D-glucose or Quip3NAc is an unusual dideoxy sugar found in the O-antigens of various Gram-negative bacteria and in the S-layer glycoprotein glycans of some Gram-positive bacteria. It is produced in these organisms as a dTDP-linked sugar, with five enzymes ultimately required for its biosynthesis. The focus of this investigation is on the enzyme QdtC, a CoA dependent N-acetyltransferase that catalyzes the last step in the Quip3NAc biosynthetic pathway. For this analysis, three crystal structures were determined: the wild-type enzyme in the presence of acetyl-CoA and two ternary complexes of the enzyme with CoA and either dTDP-D-Quip3N or dTDP-3-amino-3,6 didexoy-alpha-D-galactose (dTDP-D-Fucp3N). Each subunit of the trimeric enzyme is dominated by a left-handed beta-helix motif with 11 turns. The three active sites are located at the subunit-subunit interfaces, and the two dTDP-sugar ligands employed in this study bind to the protein in nearly identical manners. Those residues responsible for anchoring the hexose moieties of the dTDP-sugars to the protein include Glu 141, Asn 159, and Asp 160 from one subunit and His 134 from another subunit. To probe the roles of various amino acid residues in the catalytic mechanism of the enzyme, 10 site-directed mutant proteins were constructed and their kinetic parameters measured. On the basis of these data, a catalytic mechanism is proposed for QdtC in which the acetylation of the sugar amino group does not require a catalytic base provided by the protein. Rather, the sulfur of CoA functions as the ultimate proton acceptor. PMID- 19191740 TI - An enhanced molecular dynamics study of HPPK-ATP conformation space exploration and ATP binding to HPPK. AB - HPPK (6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase) catalyzes the transfer of pyrophosphate from ATP to HP (6-hydroxymethyl-7,8-dihydropterin). This first reaction in the folate biosynthetic pathway is an important target for potential antimicrobial agents. In this work, the mechanism by which HPPK traps and binds ATP is studied by molecular dynamics (MD)-based methods. Based on the ternary crystal structure of HPPK with an ATP mimic and HP, a complex of ATPMg(2) and HPPK is simulated and found to undergo small conformational changes with conventional MD, as does also conventional MD when started from the apo crystal structure. The introduction of restraints in the MD that serve to move HPPK-ATP from its ternary complex (closed) to apo-like (open) forms shows that throughout the restraint path ATP remains bound to HPPK. That ATP remains bound suggests that there is an ensemble of conformations with ATP bound to HPPK that span the apo to more ligand-bound-like conformations, consistent with the pre-existing equilibrium hypothesis of ligand binding, whereby a ligand can select from and bind to a broad range of protein conformations. In the apo-like conformations, ATPMg(2) remains bound to HPPK through a number of mainly salt-bridge-like interactions between several negatively charged residues and the two magnesium cations. The introduction of a reweight method that enhances the sampling of MD by targeting explicit terms in the force field helps define the interactions that bind ATP to HPPK. Using the reweight method, conformational and center of mass motions of ATP, driven by the breaking and making of hydrogen bonds and salt bridges, are identified that lead to ATP separating from HPPK. An elastic normal mode (ENM) approach to opening the ternary complex and closing the apo crystal structures was carried out. The ENM analysis of the apo structure analysis shows one mode that does have a closing motion of HPPK loops, but the direction does not correlate strongly with the loop motions that are required for forming the ternary complex. PMID- 19191742 TI - TSH-based protocol, tablet instability, and absorption effects on L-T4 bioequivalence. AB - BACKGROUND: FDA Guidance for pharmacokinetic (PK) testing of levothyroxine (L T(4)) for interbrand bioequivalence has evolved recently. Concerns remain about efficacy and safety of the current protocol, based on PK analysis following supraphysiological L-T(4) dosing in euthyroid volunteers, and recent recalls due to intrabrand manufacturing problems also suggest need for further refinement. We examine these interrelated issues quantitatively, using simulated what-if scenarios testing efficacy of a TSH-based protocol and tablet stability and absorption, to enhance precision of L-T(4) bioequivalence methods. METHODS: We use an updated simulation model of human thyroid hormone regulation quantified and validated from data that span a wide range of normal and abnormal thyroid system function. Bioequivalence: We explored a TSH-based protocol, using normal replacement dosing in simulated thyroidectomized patients, switching brands after 8 weeks of full replacement dosing. We simulated effects of tablet potency differences and intestinal absorption differences on predicted plasma TSH, T(4), and triiodothyronine (T(3)) dynamics. Stability: We simulated effects of potency decay and lot-by-lot differences in realistic scenarios, using actual tablet potency data spanning 2 years, comparing the recently reduced 95-105% FDA approved potency range with the original 90-110% range. RESULTS: A simulated decrease as small as 10-15% in L-T(4) or its absorption generated TSH concentrations outside the bioequivalence target range (0.5-2.5 mU/L TSH), whereas T(3) and T(4) plasma levels were maintained normal. For a 25% reduction, steady-state TSH changed 300% (from 1.5 to 6 mU/L) compared with <25% for both T(4) and T(3) (both within their reference ranges). Stability: TSH, T(4), and T(3) remained within normal ranges for most potency decay scenarios, but tablets of the same dose strength and brand were not bioequivalent between lots and between fresh and near-expired tablets. CONCLUSIONS: A pharmacodynamic TSH measurement bioequivalence protocol, using normal L-T(4) replacement dosing in athyreotic volunteers, is likely to be more sensitive and safer than current FDA Guidance based on T(4) PK. The tightened 95-105% allowable potency range for L T(4) tablets is a significant improvement, but otherwise acceptable potency differences (whether due to potency decay or lot-by-lot inconsistencies) may be problematic for some patients, for example, those undergoing high-dose L-T(4) therapy for cancer. PMID- 19191741 TI - Type 1 diabetes and autoimmune thyroiditis: the genetic connection. PMID- 19191743 TI - Mildly elevated TSH and cognition in middle-aged and older adults. AB - BACKGROUND: It is accepted that markedly elevated thyroid-stimulating hormone (TSH) levels are associated with impaired cognitive function. However, the findings regarding the association between mildly elevated TSH levels and cognition are equivocal. The objective of this study was to assess the relation between TSH levels in the normal to mildly elevated range (0.3-10.0 mIU/L) and several domains of cognitive function. METHODS: A healthy, community-based sample of 489 men and women (40-88 years old, mean = 60.5 years) enrolled in the B Vitamin Atherosclerosis Intervention Trial were studied. A neuropsychological test battery was used to assess a broad array of cognitive functions. Four uncorrelated neuropsychological factors were extracted by principal component analysis. Using multivariable linear regression, performance on each factor was examined in relation to TSH levels, controlling for age, gender, race-ethnicity, education, homocysteine levels, low-density lipoprotein cholesterol levels, and smoking status. RESULTS: TSH levels were not associated with any of the four factor scores in the total sample or in younger (age < 60) or older (age >or= 60) subjects, although there was a trend for older subjects with higher levels of TSH to do more poorly on paragraph recall (p = 0.06). Gender-stratified analyses showed that TSH was positively associated with scores on word list learning for females only (p = 0.003). CONCLUSIONS: In this community-based sample of middle aged to older individuals, increasing TSH levels were not associated with significantly reduced cognitive performance in any domain. Further exploration of the effects of gender on the association between TSH and cognition is warranted. PMID- 19191744 TI - Encapsulated papillary thyroid carcinoma: a clinico-pathologic study of 106 cases with emphasis on its morphologic subtypes (histologic growth pattern). AB - BACKGROUND: Encapsulated papillary thyroid carcinoma (EPTC) can have a histologic growth pattern similar to the one seen in classical papillary thyroid carcinoma (PTC) or akin to the follicular variant of PTC (FVPTC). This study aims to assess the behavior of EPTC according to its growth pattern. METHODS: All cases of thyroid carcinomas treated at our institution between 1980 and 2000 were reviewed and reclassified according to current histopathologic criteria. RESULTS: After review by two pathologists, 106 cases were included. Forty-three (41%) of the cases were identified as encapsulated classical PTC (E-CPTC) and 63 (59%) as encapsulated FVPTC (E-FVPTC). E-FVPTC had a higher rate of vascular invasion (16/63; 25%) than E-CPTC (2/43; 5%) (p = 0.007). In contrast, E-CPTC had a higher frequency of capsular invasion (28/43; 65%) than E-FVPTC (24/63, 38%) (p = 0.01). The lymph node metastatic rate was significantly higher in E-CPTC (11/43, 26%) compared to E-FVPTC (2/63, 3%) (p = 0.0014). All 34 noninvasive E-FVPTC lacked evidence of nodal metastases while 4 of 15 (27%) noninvasive E-CPTC presented with nodal disease (p = 0.006). Distant metastasis occurred only in four cases of E-FVPTC at presentation. These four FVPTC had extensive capsular and/or vascular invasion and no nodal disease. None of noninvasive EPTC recurred, including 30 patients treated by lobectomy without radioactive iodine (RAI) therapy (median follow-up: 8.9 years). CONCLUSION: E-CPTC resembles classical PTC in its propensity to metastasize to lymph nodes and its vascular/capsular invasive pattern while E-FVPTC behaves more like follicular carcinoma/adenoma group of tumors. Meticulous search for capsular and vascular invasion can reliably predict the metastatic potential of E-FVPTC but not of E-CPTC. The latter can therefore be treated like unencapsulated classical PTC. Noninvasive E-FVPTC could be managed like minimally invasive follicular carcinoma by lobectomy without RAI therapy. Invasive E-FVPTC seem quite indolent if no distant metastases are found at presentation. PMID- 19191745 TI - Association of XRCC1 polymorphisms and risk of differentiated thyroid carcinoma: a case-control analysis. AB - BACKGROUND: Numerous single-nucleotide polymorphisms (SNPs) of the DNA repair gene XRCC1 have been described. These SNPs have been increasingly studied in the epidemiology of various cancer types. In this study we evaluated the risk association between six common SNPs of the XRCC1 gene and differentiated thyroid carcinoma (DTC). METHODS: We conducted a case-control study of 251 subjects with DTC, 145 subjects with benign thyroid disease, and 503 cancer-free controls. Polymerase chain reaction-restriction fragment length polymorphism assays were performed for genotyping. Multivariate logistic regression analysis was performed for risk estimation. Expectation-maximization algorithm and bayesian methods were used to estimate haplotype frequencies. RESULTS: Multivariate analysis demonstrated an increased risk of DTC for the Arg194Trp heterozygous polymorphic (CT) genotype (odds ratio [OR]: 1.4, 95% confidence interval [CI]: 0.9-2.1). Multivariate analysis demonstrated a decreased risk of DTC for the Arg399Gln homozygous polymorphic (AA) genotype (OR: 0.5, 95% CI: 0.3-0.8) and the polymorphic (A) allele (OR: 0.7, 95% CI: 0.5-1.0). Compared to the most commonly observed haplotype (CGTCGA), multiple haplotypes were associated with a significantly increased risk of DTC, with the CGTTGG haplotype demonstrating the strongest association (OR: 5.0, 95% CI: 1.9-13.2). CONCLUSIONS: The XRCC1 194Trp variant allele may be associated with increased risk of DTC, while the XRCC1 399Gln variant allele may be associated with decreased risk of DTC. The utility of XRCC1 haplotypes in predicting DTC risk deserves further investigation with direct haplotype measurement. PMID- 19191746 TI - Effect of type I interferon(s) on cell viability and apoptosis in primary human thyrocyte cultures. AB - BACKGROUND: Interferon (IFN) therapy may induce a generalized activation of the immune system, hence triggering or exacerbating autoimmune disease. Apoptosis contributes to the development of hypothyroidism in autoimmune thyroiditis. IFN can affect all phases of the cell cycle and may induce apoptosis in several cell lines from varied histologies. To date, no data exist on the possible effect of type I IFN(s) on FAS/FASL system and cell apoptosis of human thyroid follicles. Therefore, we evaluated the effect of both IFN-alpha and -beta on apoptosis in primary human thyrocyte cultures and the potential role of the FAS/FASL pathway. METHODS: Thyrocytes were cultured in monolayers and FAS, FASL, and Bcl-2 mRNA expression was determined by reverse transcriptase polymerase chain reaction after exposure to 10 mIU/mL bovine thyroid-stimulating hormone alone or in combination with increasing doses of IFN-alpha or -beta for 24, 48, and 72 hours. The percentage of apoptotic hypodiploid cells was evaluated by flow cytometry. RESULTS: Thyroid-stimulating hormone significantly decreased FAS and increased Bcl-2 mRNA expression while reducing the percentage of hypodiploid cells. The concomitant addition of either IFN-alpha or -beta reduced cell viability and increased the number of hypodiloid cells, but only IFN-beta modulated the expression of FAS and Bcl-2 mRNA expression in a proapoptotic sense. CONCLUSIONS: Both type I IFN(s) increase apoptosis in primary thyrocyte cultures, but only IFN beta modulates FAS and Bcl-2 gene expression toward a proapoptotic pathway. Because apoptosis plays an important role in thyroid homeostasis and disease, this mechanism may contribute to the development and progression of type I IFN(s) therapy-associated thyroid disease. PMID- 19191747 TI - No link between X chromosome inactivation pattern and simple goiter in females: evidence from a twin study. AB - BACKGROUND: Simple goiter (SG) comprises diffuse (DG) and nodular (NG) benign nonautoimmune nontoxic goiter. In nonendemic goiter areas, the ratio of females to males may exceed 5:1, indicating that gender and/or sex hormones may play a role in the etiology of SG in these areas. Theoretically, as shown for autoimmune thyroid disease, X chromosome inactivation (XCI) and resultant tissue chimerism could offer a novel explanation for the female preponderance of SG. To examine whether skewed XCI is associated with SG, we first compared XCI in 71 twin individuals with SG with that in 142 unrelated healthy control twin individuals, and then performed a within-pair comparison of XCI in 48 twin pairs discordant for SG. METHODS: DNA was extracted from peripheral blood cells. XCI analysis was performed by predigestion of DNA using the methylation-sensitive enzyme Hpall, followed by polymerase chain reaction of the polymorphic CAG repeat of the androgen receptor gene. A polymerase chain reaction product is obtained from the inactive X chromosome only. The XCI pattern was classified as skewed when 80% or more of the cells preferentially inactivated the same X chromosome. Twin zygosity was established by DNA fingerprinting. RESULTS: The frequency of skewed XCI in female twins with SG, DG, and NG was 11% (8/71), 13% (6/46), and 8% (2/25), respectively, which was not significantly different from the prevalences in the corresponding control populations, 14% (20/142, p = 0.56), 14% (13/92, p = 1.00), and 14% (7/50, p = 0.71), respectively. Essentially, similar results were obtained when comparing the prevalence of skewed XCI in twin pairs discordant for SG (48 pairs), DG (30 pairs), and NG (18 pairs). CONCLUSION: In a sample of Danish female twins, we did not find evidence for involvement of skewed XCI in the etiology or the female preponderance of SG. PMID- 19191748 TI - A thyroid hormone deiodinase inhibitor can decrease cutaneous cell proliferation in vitro. AB - BACKGROUND: Although cutaneous manifestations associated with thyroid dysfunction are classic, the potential for thyroid hormone or its antagonists to treat dermatological disease has not been explored with rigor. The predominant circulating thyroid hormone is the pro-hormone, thyroxine (T(4)). Skin, like many tissues, expresses thyroid hormone deiodinases to convert T(4) to the active thyroid hormone, triiodothyronine (T(3)). Previously, we determined that T(3) is necessary for optimal growth of keratinocytes and fibroblasts. The first hypothesis of this experiment was that the deiodinase inhibitor iopanoic acid (IOP) could inhibit cutaneous cell proliferation. The second hypothesis of this experiment was that the action of IOP could be attributed to its inhibition of conversion of T(4) to T(3). Although IOP is known to inhibit T(4) to T(3) conversion, the inhibition of cutaneous cell proliferation by IOP might conceivably result from other properties of IOP. METHODS: In separate experiments, primary culture human keratinocytes and dermal fibroblasts were incubated overnight with IOP. RESULTS: Proliferation was inhibited in a dose dependent manner in both cell lines. Overnight incubation with T(3) restored the proliferation but overnight incubation with T(4) did not. CONCLUSION: The study is the first to demonstrate that IOP inhibits cutaneous cell proliferation and that supplemental T(3) is sufficient to correct at least part of the inhibition caused by IOP. The data suggest that IOP decreases cutaneous cell proliferation by inhibition of intracellular T(4) to T(3) conversion. The data provide indirect evidence of the presence of type 1 or type 2 iodothyronine deiodinase activity in skin cells. The data support our previous hypothesis that T(3) is necessary for normal cutaneous proliferation. PMID- 19191749 TI - A somatic gain-of-function mutation in the thyrotropin receptor gene producing a toxic adenoma in an infant. AB - BACKGROUND: Activating mutations of the thyroid stimulating hormone receptor gene (TSHR) are rare in the neonate and in the pediatric population. They are usually present in the germline, and are either inherited or occur de novo. Somatic mutations in TSHR are unusual in the pediatric population. METHODS: We describe a nine-month-old infant with thyrotoxicosis who harbored an activating somatic mutation in TSHR that was not present in the germline. RESULTS: As genomic DNA analysis failed to show a TSHR gene mutation, a radioiodide scan was performed to reveal a unilateral localization of uptake suppressing the remaining thyroid tissue. Genomic and complementary DNA analyses of the active thyroid tissue, removed surgically, identified a missense mutation (D633Y) located in the sixth transmembrane domain of the TSHR. The absence of this TSHR mutation in circulating mononuclear cells and in unaffected thyroid tissue confirmed the somatic nature of this genetic alteration. CONCLUSIONS: To the authors' knowledge, this is the youngest patient to receive definitive treatment for hyperthyroidism due to an activating mutation of TSHR. PMID- 19191750 TI - Emergency dilation by self-expandable tracheal stent for upper airway obstruction in a patient with a giant primary thyroid lymphoma. PMID- 19191751 TI - Bilateral breast involvement in medullary thyroid carcinoma. PMID- 19191752 TI - A ganglioneuroma with features of a thyroid nodule: intense pain on fine needle biopsy as a diagnostic clue. PMID- 19191757 TI - Action of RANKL and OPG for osteoclastogenesis. AB - Osteoclasts develop from hematopoietic cells of the monocyte-macrophage lineage. The coculture system of osteoblasts and hematopoietic cells was devised to examine osteoclastogenesis in vitro. Experiments using the coculture system have established the concept that osteoblasts are crucially involved in osteoclastogenesis. Remarkable progress has been achieved during the last decade in our understanding the molecular mechanism of osteoclast differentiation, largely because of the discovery of receptor activator of NF-kappaB ligand (RANKL), an essential cytokine for osteoclastogenesis. Osteoblasts express RANKL in response to bone-resorbing factors. Osteoblasts also produce osteoprotegerin (OPG), a decoy receptor for RANKL, which inhibits osteoclast differentiation and function by interrupting the interaction between RANKL and RANK, a receptor of RANKL. The identification of nuclear factor of activated T-cells cytoplasmic 1 (NFATc1) as a master transcription! factor for RANKL-induced osteoclastogenesis has provided major insight into the molecular mechanism of osteoclast differentiation. The discovery of the immunoreceptor tyrosine-based activation motif (ITAM)-mediated signals as a costimulatory signal in osteoclastogenesis has confirmed that osteoblasts play another important role in osteoclastogenesis. Mutations of RANK, OPG, and RANKL found in humans cause bone diseases associated with expected skeletal abnormalities. Thus, the RANKL/RANK/OPG axis is now recognized as the central regulator of osteoclast differentiation and function. PMID- 19191758 TI - Molecular mechanisms in coupling of bone formation to resorption. AB - Bone remodeling is the process of removal and replacement of bone, taking place at many sites throughout the skeleton and regulated mainly by locally generated factors. Its purposes are to repair damaged bone, remove old bone, and facilitate skeletal responses to changes in loading requirements. Cells of the osteoblast lineage control the formation and activity of osteoclasts, which are responsible for initiation and execution of resorption at remodeling sites. The bone resorbed by osteoclasts is replaced through the differentiation and activity of osteoblasts. The consequent formation must match closely the amount of bone that is resorbed at each site. This coupling of the two processes is essential for bone balance. Both resorption products and osteoclast-derived factors contribute to the coupling of bone formation to resorption in bone remodeling. This review considers the molecular mechanisms and intercellular communication processes involved in remodeling and coupling. PMID- 19191759 TI - Cholesteryl ester transfer protein (CETP) increases postprandial triglyceridaemia and delays triacylglycerol plasma clearance in transgenic mice. AB - The CETP (cholesteryl ester transfer protein) is a plasma protein synthesized in several tissues, mainly in the liver; CETP reduces plasma HDL (high-density lipoprotein) cholesterol and increases the risk of atherosclerosis. The effect of CETP levels on postprandial intravascular metabolism of TAGs (triacylglycerols) is an often-overlooked aspect of the relationship between CETP and lipoprotein metabolism. Here, we tested the hypothesis that CETP delays the plasma clearance of TAG-rich lipoprotein by comparing human CETP expressing Tg (transgenic) and non-Tg mice. After an oral fat load, the postprandial triglyceridaemia curve was markedly increased in CETP-Tg compared with non-Tg mice (280+/-30 versus 190+/-20 mg/dl per 6 h respectively, P<0.02). No differences in intestinal fat absorption and VLDL (very-low-density lipoprotein) secretion rates were observed. Kinetic studies of double-labelled chylomicron-like EMs (emulsions) showed that both [(3)H]triolein and [(14)C]cholesteryl oleate FCRs (fractional clearance rates) were significantly reduced ( approximately 20%) in CETP-Tg mice. Furthermore, TAG from lipid EM pre-incubated with CETP-Tg plasma had plasma clearance and liver uptake significantly lower than the non-Tg plasma-treated lipid EM. In addition, reductions in post-heparin plasma LPL (lipoprotein lipase) activity (50%) and adipose tissue mRNA abundance (39%) were verified in CETP-Tg mice. Therefore we conclude that CETP expression in Tg mice delays plasma clearance and liver uptake of TAG-rich lipoproteins by two mechanisms: (i) transferring TAG to HDLs and increasing CE content of the remnant particles and (ii) by diminishing LPL expression. These findings show that the level of CETP expression can influence the responsiveness to dietary fat and may lead to fat intolerance. PMID- 19191756 TI - Cancer treatment-related bone disease. AB - Bone health may be impaired in many patients being treated for cancer. Primary tumors that reside in or form metastases to bone can result in compromised skeletal integrity. It has also been increasingly recognized that patients undergoing therapies for treatment of cancer are at higher risk of bone loss. These include androgen-deprivation therapy for prostate cancer and aromatase inhibitor therapy for breast cancer, among others. Hypogonadism induced by many of these cancer treatments results in bone loss and increases the risk of osteoporosis and fractures. Progress has been made in identifying the role of oral and intravenous bisphosphonates to prevent bone loss in these patients. This review discusses bone loss associated with cancer treatments, with a focus on breast cancer, prostate cancer, and survivors of childhood malignancies. PMID- 19191760 TI - Expression of the iron-regulatory protein haemojuvelin in retina and its regulation during cytomegalovirus infection. AB - Haemochromatosis is a genetic disorder of iron overload resulting from loss-of function mutations in genes coding for the iron-regulatory proteins HFE [HLA-like protein involved in iron (Fe) homoeostasis], transferrin receptor 2, ferroportin, hepcidin and HJV (haemojuvelin). Expression of the first four genes coding for these proteins in retina has been established. Here we report on the expression of HJV. Since infection of retina with CMV (cytomegalovirus) causes blindness, we also investigated the expression of HJV and other iron-regulatory proteins in retina during CMV infection. HJV (HJV gene) mRNA was expressed in RPE (retinal pigment epithelium)/eyecup and neural retina in mouse. In situ hybridization and immunohistochemistry confirmed the presence of HJV mRNA and protein in RPE, outer and inner nuclear layers, and ganglion cell layer. Immunocytochemistry with cell lines and primary cell cultures showed HJV expression in RPE and Muller cells. In RPE, the expression was restricted to apical membrane. Infection of primary cultures of mouse RPE with CMV increased HJV mRNA and protein levels. Under similar conditions, HFE (HFE gene) mRNA levels were not altered, but HFE protein was decreased. Hepcidin expression was, however, not altered. These findings were demonstrable in vivo with CMV-infected mouse retina. The CMV-induced up regulation of HJV in RPE was independent of changes in HFE because the phenomenon was also seen in HFE-null RPE cells. CMV-infected primary RPE cells showed evidence of iron accumulation and oxidative stress, as indicated by increased levels of ferritin and hydroxynonenal. The observed changes in HJV expression and iron status during CMV infection in retina may have significance in the pathophysiology of CMV retinitis. PMID- 19191761 TI - Helicobacter pylori, helminth infections and growth: a cross-sectional study in a high prevalence population. AB - AIM: Helicobacter pylori (H. pylori) infection is usually acquired in childhood and may adversely affect growth, although data are inconsistent. This study investigated growth parameters in a paediatric cohort with a high prevalence of H. pylori and helminth infection. METHODS: A cross-sectional study of African refugee children (<16 years) recruited at their initial health assessment following resettlement in Australia. Detailed demographic, infection and anthropometric data were obtained. H. pylori infection was diagnosed by monoclonal faecal antigen enzyme immunoassay testing (MFAT). Growth restriction was defined as children with any anthropometric measures below the fifth centile for age and gender. RESULTS: H. pylori infection was detected in 149/182 (81.9%). Children with H. pylori infection were older (mean 8.5 years, standard deviation (SD) 4.2 years vs. 5.8 years, SD 4.5 years, p < 0.001). No gender differences were observed. After adjustment for age, H. pylori did not adversely affect body mass index or other anthropometric measurements. Helminth infections were common (41.8%) but not associated with reduced growth or with H. pylori infection. CONCLUSION: H. pylori and helminth infections are prevalent in African refugee children but neither is associated with growth restriction. Longitudinal growth velocity studies are necessary to identify any long-term consequences of H. pylori on childhood growth. PMID- 19191762 TI - Adverse neurodevelopmental outcome in preterm infants: risk factor profiles for different gestational ages. AB - AIM: Assessment of risk predictors for adverse neurodevelopmental outcome at 1 year of age in preterm infants with a gestational age <30 weeks (Group I) and 30 32 weeks (Group II). METHODS: Between January 2003 and December 2006, we prospectively enrolled 310 live-born infants between 23 and 32 weeks of gestation. The association between candidate risk factors and delayed motor or mental development (Bayley Scales of infant development II; psychomotor or mental developmental index <85) was analysed by means of logistic regression analysis. RESULTS: Two hundred and fifty infants were eligible for follow-up, and 205 (82.0%) completed the follow-up visit. Intracerebral haemorrhage, small for gestational age and late-onset sepsis were associated with an increased risk for delayed development in Group I (p < 0.05, each). Premature rupture of membranes was a risk condition relevant to Group II. Antenatal steroids were associated with a decreased risk of neurodevelopmental delay in both groups. CONCLUSION: This study identified distinct risk factors for adverse outcome in preterm infants of lower (<30 weeks) and higher (30-32 weeks) gestational age. In the lower gestational age group, neonatal risk predictors are most important. Antenatal steroids appear to decrease the risk for adverse outcome in both age groups. PMID- 19191755 TI - Signaling networks that control the lineage commitment and differentiation of bone cells. AB - Osteoblasts and osteoclasts are the two major bone cells involved in the bone remodeling process. Osteoblasts are responsible for bone formation while osteoclasts are the bone-resorbing cells. The major event that triggers osteogenesis and bone remodeling is the transition of mesenchymal stem cells into differentiating osteoblast cells and monocyte/macrophage precursors into differentiating osteoclasts. Imbalance in differentiation and function of these two cell types will result in skeletal diseases such as osteoporosis, Paget's disease, rheumatoid arthritis, osteopetrosis, periodontal disease, and bone cancer metastases. Osteoblast and osteoclast commitment and differentiation are controlled by complex activities involving signal transduction and transcriptional regulation of gene expression. Recent advances in molecular and genetic studies using gene targeting in mice enable a better understanding of the multiple factors and signaling networks that control the differentiation process at a molecular level. This review summarizes recent advances in studies of signaling transduction pathways and transcriptional regulation of osteoblast and osteoclast cell lineage commitment and differentiation. Understanding the signaling networks that control the commitment and differentiation of bone cells will not only expand our basic understanding of the molecular mechanisms of skeletal development but will also aid our ability to develop therapeutic means of intervention in skeletal diseases. PMID- 19191763 TI - Effect of comorbidity adjustment on CMS criteria for kidney transplant center performance. AB - The Centers for Medicare & Medicaid Services (CMS) uses kidney transplant outcomes, unadjusted for standard comorbidity, to identify centers with sufficiently higher than expected rates of graft failure or patient death (underperforming centers) that they may be denied Medicare participation. To examine whether comorbidity adjustment would affect this determination, we identified centers that would have failed to meet 1-year graft survival criteria, 1992-2005, with and without adjustment using the Elixhauser Comorbidity Index. Adjustment was performed for each U.S. center for 24 consecutive (overlapping) 30 month intervals, including 102 176 adult deceased-donor and living-donor kidney transplant patients with Medicare as primary payer 6 months pretransplant. For each interval, we determined percent positive agreement (PPA) (number of centers underperforming both before and after adjustment, divided by number underperforming either before or after adjustment). Overall PPA was 80.8%, with no evidence of a trend over time. Among deceased-donor recipients, 10 of 31 comorbid conditions were predictors of graft failure in at least half of the intervals, as were six conditions among living-donor recipients. Lack of comorbidity adjustment may disadvantage centers willing to accept higher risk patients. Risk of jeopardizing Medicare funding may give centers incentive to deny transplantation to higher risk patients. PMID- 19191764 TI - Positive cross-match living donor kidney transplantation: longer-term outcomes. AB - The long-term graft outcomes after positive cross-match (PXM) living donor kidney transplantation (LDKT) are unknown and the descriptive published data present short-medium term results. We conducted a retrospective cohort study of LDKT with PXM by flow cytometry performed at our center during February 1999 to October 2006, compared to a control group, matched 1:1 for age, sex, race, retransplantation and transplant year. The PXM group was treated with a course of plasmapheresis/low-dose intravenous immunoglobulin (IVIg) preoperatively, and OKT3 or thymoglobulin induction. Both groups (n = 41 each) were comparable except for duration of end-stage renal disease (ESRD), induction, HLA mismatch and panel reactive antibody (PRA). During the period of up to 9 years, 14 PXM and 7 controls lost their grafts (p < 0.04). Graft survival rates at 1 and 5 years were 89.9% and 69.4% for PXM group and 97.6% and 80.6% for the controls, respectively. PXM was associated with higher risk of graft loss (HR 2.6, p = 0.04; 95%CI 1.03 6.4) (t(1/2)= 6.8 years), but not with patient survival (HR 1.96, p = 0.29; 95%CI 0.6-7.0) or 1-year serum creatinine (beta= 0.06, p = 0.59 for ln (SCr); 95% CI 0.16 to 0.28). These results suggest that despite the favorable short-term results of PXM LDKT after PP/IVIg conditioning, medium-long-term outcomes are notably worse than expected, perhaps comparable to non-ECD deceased donor kidney transplantation (KT). PMID- 19191765 TI - Formal policies and special informed consent are associated with higher provider utilization of CDC high-risk donor organs. AB - A new United Network for Organ Sharing (UNOS) policy mandates special informed consent (SIC) before transplanting organs from donors classified by the Public Health Service/Center for Disease Control (PHS/CDC) as high-risk donors (HRDs); however, concerns remain that this policy may cause suboptimal organ utilization. Currently, consent and disclosure policy is determined by individual centers or surgeons; as such, little is known about current practices. The goals of this study were to quantify consent and disclosure practices for HRDs in the United States, identify factors associated with SIC use and analyze associations between SIC use and HRD organ utilization. We surveyed 422 transplant surgeons about their use of HRD organs and their associated consent and disclosure practices. In total, 52.7% of surgeons use SIC, but there is a high variation in use within centers, between centers and by donor behavior. A defined HRD policy at a transplant center is strongly associated with SIC use at that center (OR = 4.68, p < 0.001 by multivariate hierarchical logistic regression). SIC use is associated with higher utilization of HRD livers (OR 3.37), and a trend toward higher utilization of HRD kidneys (OR 1.74) and pancreata (OR 1.28). We believe our findings support a formalized national policy and suggest that this policy will not result in decreased utilization. PMID- 19191766 TI - Viral nucleic acid testing (NAT) and OPO-level disposition of high-risk donor organs. AB - The use of Public Health Service/Centers for Disease Control and Prevention (PHS/CDC) high-risk donor (HRD) organs remains controversial, especially in light of a recent high-profile case of human immunodeficiency virus (HIV) and hepatitis C virus (HCV) transmission. Nucleic acid testing (NAT), while more expensive and time consuming, reduces infectious risk by shortening the period between infection and detectability. The purpose of this study was to characterize HRDs and disposition of their organs by organ procurement organization (OPO), to measure NAT practices by OPO and to examine associations between NAT practices and use of HRD organs. We analyzed 29 950 deceased donors (2574 HRDs) reported to UNOS since July 1, 2004 and May 8, 2008. We then surveyed all OPO clinical directors about their use of NAT, average time to receive NAT results, locations where NAT is performed and percentage of the time NAT results are available for allocation decisions. In total, 51.7% of OPOs always perform HIV NAT, while 24.1% never do. A similar pattern is seen for HCV NAT performance, while the majority (65.6%) never perform HBV NAT. AIDS prevalence in an OPO service area is not associated with NAT practice. OPOs that perform HIV NAT are less likely to export organs outside of their region. The wide variation of current practice and the possibility that NAT would improve organ utilization support consideration for a national policy. PMID- 19191767 TI - Therapeutic targets in liver transplantation: angiotensin II in nonsteatotic grafts and angiotensin-(1-7) in steatotic grafts. AB - Numerous steatotic livers are discarded as unsuitable for transplantation because of their poor tolerance of ischemia-reperfusion(I/R). The injurious effects of angiotensin (Ang)-II and the benefits of Ang-(1-7) in various pathologies are well documented. We examined the generation of Ang II and Ang-(1-7) in steatotic and nonsteatotic liver grafts from Zucker rats following transplantation. We also studied in both liver grafts the effects of Ang-II receptors antagonists and Ang (1-7) receptor antagonists on hepatic I/R damage associated with transplantation. Nonsteatotic grafts showed higher Ang II levels than steatotic grafts, whereas steatotic grafts showed higher Ang-(1-7) levels than nonsteatotic grafts. Ang II receptor antagonists protected only nonsteatotic grafts against damage, whereas Ang-(1-7) receptor antagonists were effective only in steatotic grafts. The protection conferred by Ang II receptor antagonists in nonsteatotic grafts was associated with ERK 1/2 overexpression, whereas the beneficial effects of Ang-(1 7) receptor antagonists in steatotic grafts may be mediated by NO inhibition. Our results show that Ang II receptor antagonists are effective only in nonsteatotic liver transplantation and point to a novel therapeutic target in liver transplantation based on Ang-(1-7), which is specific for steatotic liver grafts. PMID- 19191768 TI - Serum autoantibodies against cytochrome P450 2E1 (CYP2E1) predict severity of necroinflammation of recurrent hepatitis C. AB - We previously reported that autoantibodies against cytochrome P4502E1 (CYP2E1) are frequent in patients with chronic hepatitis C. As autoimmune reactions are increasingly detected after orthotopic liver transplantation (OLT), this study investigates prevalence and significance of anti-CYP2E1 autoantibodies in 46 patients with post-OLT recurrent hepatitis C. IgG against recombinant human CYP2E1 above the control threshold was detected in 19 out 46 (41%) sera collected immediately before OLT and in 15 out 46 (33%) sera collected at the time of the 12 months follow-up liver biopsy. Although anti-CYP2E1 reactivity was not modified by OLT, the patients with persistently elevated anti-CYP2E1 IgG (n = 12; 26%) showed significantly higher prevalence of recurrent hepatitis with severe necroinflammation and fibrosis than those persistently negative or positive only either before or after OLT. Moreover, the probability of developing severe necroinflammation was significantly higher in persistently anti-CYP2E1-positive subjects. Multivariate regression and Cox analysis confirmed that the persistence of anti-CYP2E1 IgG, together with a history of acute cellular rejection and donor age >50 years, was an independent risk factor for developing recurrent hepatitis C with severe necroinflammation. We propose that autoimmune reactions involving CYP2E1 might contribute to hepatic damage in a subgroup of transplanted patients with recurrent hepatitis C. PMID- 19191769 TI - Identifying specific causes of kidney allograft loss. AB - The causes of kidney allograft loss remain unclear. Herein we investigated these causes in 1317 conventional kidney recipients. The cause of graft loss was determined by reviewing clinical and histologic information the latter available in 98% of cases. During 50.3 +/- 32.6 months of follow-up, 330 grafts were lost (25.0%), 138 (10.4%) due to death with function, 39 (2.9%) due to primary nonfunction and 153 (11.6%) due to graft failure censored for death. The latter group was subdivided by cause into: glomerular diseases (n = 56, 36.6%); fibrosis/atrophy (n = 47, 30.7%); medical/surgical conditions (n = 25, 16.3%); acute rejection (n = 18, 11.8%); and unclassifiable (n = 7, 4.6%). Glomerular pathologies leading to failure included recurrent disease (n = 23), transplant glomerulopathy (n = 23) and presumed nonrecurrent disease (n = 10). In cases with fibrosis/atrophy a specific cause(s) was identified in 81% and it was rarely attributable to calcineurin inhibitor (CNI) toxicity alone (n = 1, 0.7%). Contrary to current concepts, most cases of kidney graft loss have an identifiable cause that is not idiopathic fibrosis/atrophy or CNI toxicity. Glomerular pathologies cause the largest proportion of graft loss and alloinmunity remains the most common mechanism leading to failure. This study identifies targets for investigation and intervention that may result in improved kidney transplantation outcomes. PMID- 19191770 TI - Follicular bronchiolitis: a rare cause of bronchiolitis obliterans syndrome after lung transplantation: a case report. AB - This case report is the first confirmed case of follicular bronchiolitis (FB), a rare bronchiolar disorder characterized by peribronchiolar lymphoid follicles, in a series of over 400 lung transplantations performed in our center. It is to our knowledge, the first publication describing FB after lung transplantation (LTx), presenting as chronic allograft dysfunction or bronchiolitis obliterans syndrome (BOS). PMID- 19191772 TI - Renal allografts with IF/TA display distinct expression profiles of metzincins and related genes. AB - Chronic renal allograft injury is often reflected by interstitial fibrosis (IF) and tubular atrophy (TA) without evidence of specific etiology. In most instances, IF/TA remains an irreversible disorder, representing a major cause of long-term allograft loss. As members of the protease family metzincins and functionally related genes are involved in fibrotic and sclerotic processes of the extracellular matrix (ECM), we hypothesized their deregulation in IF/TA. Gene expression and protein level analyses using allograft biopsies with and without Banff'05 classified IF/TA illustrated their deregulation. Expression profiles of these genes differentiated IF/TA from Banff'05 classified Normal biopsies in three independent microarray studies and demonstrated histological progression of IF/TA I to III. Significant upregulation of matrix metalloprotease-7 (MMP-7) and thrombospondin-2 (THBS-2) in IF/TA biopsies and sera was revealed in two independent patient sets. Furthermore, elevated THBS-2, osteopontin (SPP1) and beta-catenin may play regulatory roles on MMP. Our findings further suggest that deregulated ECM remodeling and possibly epithelial to mesenchymal transition (EMT) are implicated in IF/TA of kidney transplants, and that metzincins and related genes play an important role in these processes. Profiling of these genes may be used to complement IF/TA diagnosis and to disclose IF/TA progression in kidney transplant recipients. PMID- 19191771 TI - 18F-FDG-uptake of hepatocellular carcinoma on PET predicts microvascular tumor invasion in liver transplant patients. AB - Vascular invasion of hepatocellular carcinoma (HCC) is a major risk factor for poor outcome after liver transplantation (LT). The aim of this retrospective analysis was to assess the value of preoperative positron emission tomography (PET) using (18)F-fluorodeoxyglucose ((18)F-FDG) in liver transplant candidates with HCC for predicting microvascular tumor invasion (MVI) and posttransplant tumor recurrence. Forty-two patients underwent LT for HCC after PET evaluation. Sixteen patients had an increased (18)F-FDG tumor uptake on preoperative PET scans (PET +), while 26 recipients revealed negative PET findings (PET-) pre-LT. PET- recipients demonstrated a significantly better 3-year recurrence-free survival (93%) than PET + patients (35%, p < 0.001). HCC recurrence rate was 50% in the PET + group, and 3.8% in the PET-population (p < 0.001). PET + status was identified as independent predictor of MVI [hazard ratio: 13.4]. Patients with advanced PET negative tumors and patients with HCC meeting the Milan criteria had a comparable 3-year-recurrence-free survival (80% vs. 94%, p = 0.6). Increased (18)F-FDG uptake on PET is predictive for MVI and tumor recurrence after LT for HCC. Its application may identify eligible liver transplant candidates with tumors beyond the Milan criteria. PMID- 19191773 TI - Resource utilization of living donor versus deceased donor liver transplantation is similar at an experienced transplant center. AB - Although living donor liver transplantation (LDLT) has been shown to decrease waiting-list mortality, little is known of its financial impact relative to deceased donor liver transplantation (DDLT). We performed a retrospective cohort study of the comprehensive resource utilization, using financial charges as a surrogate measure-from the pretransplant through the posttransplant periods-of 489 adult liver transplants (LDLT n = 86; DDLT n = 403) between January 1, 2000, through December 31, 2006, at a single center with substantial experience in LDLT. Baseline characteristics differed between LDLT versus DDLT with regards to age at transplantation (p = 0.02), male gender (p < 0.01), percentage Caucasians (p < 0.01) and transplant model for end-stage liver disease (MELD) score (p < 0.01). In univariate analysis, there was a trend toward decreased total transplant charges with LDLT (p = 0.06), despite increased surgical charges associated with LDLT (p < 0.01). After adjustment for the covariates that were associated with financial charges, there was no significant difference in total transplant charges (p = 0.82). MELD score at transplant was the strongest driver of resource utilization. We conclude that at an experienced transplant center, LDLT imposes a similar overall financial burden than DDLT, despite the increased complexity of living donor surgery and the addition of the costs of the living donor. We speculate that LDLT optimizes transplantation by transplanting healthier and younger recipients. PMID- 19191774 TI - Stability of alemtuzumab for low-dose induction and test doses. PMID- 19191775 TI - Functional endoscopic sinus surgery improved asthma symptoms as well as PEFR and olfaction in patients with nasal polyposis. AB - BACKGROUND: Nasal polyposis is a disease known to be associated with asthma. The management is anti-inflammatory, with topical and oral corticosteroids as the first-line treatment. The effect of surgical treatment on lower airway inflammation has not been sufficiently studied. AIM: The aim of this study is to investigate the effects of functional endoscopic sinus surgery (FESS) as well as fluticasone proprionate nasal drops (FPND) 400 microg b.i.d. on nasal and lower airway parameters in asthmatics with nasal polyposis. METHODS: This was a prospective 21-week study of 68 patients with asthma and nasal polyposis, on the benefits of FESS on nasal '(butanol test, subjective olfaction, peak nasal inspiratory flow, congestion, rhinorrhoea, and polyp score)', and on the lower airway parameters (dyspnea, cough, mean daily peak expiratory flow rate (PEFR), and lung function tests). It also included a randomized, double-blind, placebo controlled 14 weeks phase on FPND. RESULTS: Functional endoscopic sinus surgery significantly improved mean asthma symptom scores and daily PEFR and all nasal parameters including subjective and objective olfaction tests. This is the first study that shows the benefits of FESS on butanol tests in patients with nasal polyposis. We found no significant difference between topical treatment with FPND or placebo in the nasal or lower airway variables. CONCLUSION: Functional endoscopic sinus surgery improved nasal and asthma symptoms in patients with nasal polyposis. Functional endoscopic sinus surgery could be considered early in the natural course of nasal polyposis with concomitant asthma, as well as a second-line treatment in nasal polyposis patients with a reduced sense of smell. The potential benefits of FPND 400 microg b.i.d. were probably overshadowed by FESS. PMID- 19191776 TI - Indications for induction of labour: a best-evidence review. AB - BACKGROUND: Rates of labour induction are increasing. OBJECTIVES: To review the evidence supporting indications for induction. SEARCH STRATEGY: We listed indications for labour induction and then reviewed the evidence. We searched MEDLINE and the Cochrane Library between 1980 and April 2008 using several terms and combinations, including induction of labour, premature rupture of membranes, post-term pregnancy, preterm prelabour rupture of membranes (PROM), multiple gestation, suspected macrosomia, diabetes, gestational diabetes mellitus, cardiac disease, fetal anomalies, systemic lupus erythematosis, oligohydramnios, alloimmunization, rhesus disease, intrahepatic cholestasis of pregnancy (IHCP), and intrauterine growth restriction (IUGR). We performed a review of the literature supporting each indication. SELECTION CRITERIA: We identified 1387 abstracts and reviewed 418 full text articles. We preferentially included high quality systematic reviews or large randomised trials. Where no such studies existed, we included the best evidence available from smaller randomised trials and observational studies. MAIN RESULTS: We included 34 full text articles. For each indication, we assigned levels of evidence and grades of recommendation based upon the GRADE system. Recommendations for induction of labour for post term gestation, PROM at term, and premature rupture of membranes near term with pulmonary maturity are supported by the evidence. Induction for IUGR before term reduces intrauterine fetal death, but increases caesarean deliveries and neonatal deaths. Evidence is insufficient to support induction for women with insulin requiring diabetes, twin gestation, fetal macrosomia, oligohydramnios, cholestasis of pregnancy, maternal cardiac disease and fetal gastroschisis. AUTHORS' CONCLUSIONS: Research is needed to determine risks and benefits of induction for many commonly advocated clinical indications. PMID- 19191777 TI - Occurrence of placental abruption in relatives. AB - OBJECTIVE: The aim of this study was to assess the recurrence of placental abruption by severity, comparing the risk in a woman with that of recurrence in her sister and in the partner of her brother. DESIGN: Prospective observational study. SETTING: General population. POPULATION: Population-based study based on records of pregnancies from the Medical Birth Registry of Norway; 377.902 sisters with 767 395 pregnancies, 168,142 families incorporating 2-10 sisters, and 346,385 brothers with 717,604 pregnancies in their partners were identified. METHODS: Placental abruption with preterm birth, birthweight below 2500 g or perinatal death was defined as severe, other cases as mild. Because of the nested family data structure, multilevel multivariate regression was used. MAIN OUTCOME MEASURES: Placental abruption (severe and mild). RESULTS: Adjusted odds ratios of recurrence of mild and severe abruption were 6.5 (1.7%) and 11.5 (3.8%), respectively, compared with risks of 0.2 and 0.3% in the total population. After a severe abruption, odds ratios in her sisters were 1.7-2.1, whereas mild abruption produced no increased recurrence in sisters. The estimated heritability between sisters of severe abruption was 16%. No excess rate of abruption was observed between sisters and brothers' partners, between brothers' partners, or from brothers' partners to sisters. The odds ratios for a third abruption after a second abruption and a second severe abruption were 38.7 (19%) and 50.1 (24%), respectively. CONCLUSIONS: The recurrence risk of placental abruption in the same woman was higher after severe than mild abruption. Severe abruption was associated with a two-fold risk in sisters. Pregnancies following a second abruption should be considered very high risk. PMID- 19191778 TI - Optimal management strategies for placenta accreta. AB - OBJECTIVE: To determine which interventions for managing placenta accreta were associated with reduced maternal morbidity. DESIGN: Retrospective cohort study. SETTING: Two tertiary care teaching hospitals in Utah. POPULATION: All identified cases of placenta accreta from 1996 to 2008. METHODS: Cases of placenta accreta were identified using standard ICD-9 codes for placenta accreta, placenta praevia, and caesarean hysterectomy. Medical records were then abstracted for maternal medical history, hospital course, and maternal and neonatal outcomes. Maternal and neonatal complications were compared according to antenatal suspicion of accreta, indications for delivery, preoperative preparation, attempts at placental removal before hysterectomy, and hypogastric artery ligation. MAIN OUTCOME MEASURES: Early morbidity (prolonged maternal intensive care unit admission, large volume of blood transfusion, coagulopathy, ureteral injury, or early re-operation) and late morbidity (intra-abdominal infection, hospital re-admission, or need for delayed re-operation). Results Seventy-six cases of placenta accreta were identified. When accreta was suspected, scheduled caesarean hysterectomy without attempting placental removal was associated with a significantly reduced rate of early morbidity compared with cases in which placental removal was attempted (67 versus 36%, P=0.038). Women with preoperative bilateral ureteric stents had a lower incidence of early morbidity compared with women without stents (18 versus 55%, P=0.018). Hypogastric artery ligation did not reduce maternal morbidity. CONCLUSIONS: Scheduled caesarean hysterectomy with preoperative ureteric stent placement and avoiding attempted placental removal are associated with reduced maternal morbidity in women with suspected placenta accreta. PMID- 19191779 TI - Prospective randomised trial comparing gonadotrophin-releasing hormone analogues with triple tourniquets at open myomectomy. AB - OBJECTIVE: To compare intra-operative blood loss with triple tourniquets to occlude uterine blood supply against preoperative treatment with gonadotrophin releasing hormone (GnRH) analogues at open myomectomy. DESIGN: A prospective randomised controlled trial. SETTING: University teaching hospital. POPULATION: Forty women undergoing open myomectomy for symptomatic fibroids. METHODS: Women due to undergo open myomectomy were randomised to either 3 months pre-treatment with a GnRH analogue or the intra-operative application of triple tourniquets (number 1 polyglactin suture [Vicryl Ethicon Inc., Somerville, NJ, USA] tied around the cervix and a size 10 polythene suction catheter tied around the infundibulo-pelvic ligaments) to occlude the uterine blood supply. MAIN OUTCOME MEASURES: The primary outcome measure was intra-operative blood loss. Secondary outcome measures included postoperative blood loss, blood transfusion rate and postoperative morbidity. RESULTS: The two groups were similar in baseline characteristics. An average of 15 and 22 fibroids were removed from the GnRH analogue and tourniquet groups respectively. Intra-operative estimated blood loss was significantly higher in the GnRH analogue group (median 2482 ml, 75% percentile 1744-3151) than when triple tourniquets were used (median 640 ml, 75% percentile 418-881), giving a difference between means of 1842 ml (P<0.001). Similarly, significantly more women required blood transfusion in the GnRH analogue group (70 versus 30%, P<0.025). Postoperative morbidity was similar between the two groups. There were two serious complications in the tourniquet group, but they were not considered to be directly related to occlusion of the uterine blood supply. CONCLUSIONS: Triple tourniquets are significantly more effective than preoperative treatment with GnRH analogues at reducing intra operative blood loss at open myomectomy. PMID- 19191780 TI - Perineal approach to vascular anatomy during transobturator cystocele repair. AB - OBJECTIVE: To evaluate the risk of vascular injury during transobturator approach of cystocele repair. DESIGN: Dissection of the obturator area by perineal approach was performed after placement of mesh needles used for cystocele mesh repair. SETTING: Surgery school of Paris. POPULATION OR SAMPLE: Twenty obturator regions in ten fresh female cadavers. METHODS: Transperineal dissection of the obturator area was conducted in ten fresh female anatomic subjects after inserting anterior Prolift needles. MAIN OUTCOME MEASURES: The vascular anatomy of the obturator region was mapped. Distances between needles and vascular structures of the obturator area were measured three times and averaged for each side. RESULTS: The anterior cannula-equipped needle perforated the gracilis and the adductor brevis muscles. The mean (SD) distance to the anterior obturator vessels was 21.2 (1.6) mm on the right side and 20.4 (1.5) mm on the left. The posterior needle perforated the adductor magnus. Its distance to the posterior division of the obturator vessels was 1.8 (1.0) mm on the right side and 1.1 (0.9) mm on the left. CONCLUSIONS: During mesh cystocele repair by transobturator approach, the posterior obturator vessels division seems at risk of injury during the posterior needle insertion. PMID- 19191781 TI - Epistaxis during treatment with paclitaxel. AB - The purpose of this study was to determine the incidence and severity of epistaxis in patients treated with paclitaxel. Patients who were treated with paclitaxel filled a questionnaire regarding their general health, medications and incidents of epistaxis. Relevant clinical information was obtained from the patients' charts. Forty-seven consecutive patients were recruited to the study. Twenty-four (51%) of the patients reported epistaxis during paclitaxel therapy. Twenty-three of 39 (59%) patients who received weekly paclitaxel had epistaxis at least once during treatment, compared with one out of eight patients who were treated every 3 weeks (P = 0.045). All episodes of epistaxis were mild, occurred with normal platelets counts and did not require blood product transfusions or treatment modification. The majority of the patients experienced the first episode of epistaxis on the third week of weekly paclitaxel treatment and then repeatedly throughout therapy. It is concluded that epistaxis is a common mild side-effect of weekly paclitaxel that has not been reported previously. In this trial, epistaxis did not have any major clinical consequences. However, when paclitaxel is combined with other drugs that may cause bleeding, such as bevacizumab, physicians should be alerted to the potential risk of this phenomenon. PMID- 19191782 TI - Derivatives of prostate-specific antigen as predictors of incidental prostate cancer. AB - OBJECTIVE To search for an optimal derivative of prostate-specific antigen (PSA) identifying patients at risk of incidental prostate cancer. PATIENTS AND METHODS In all, 693 patients who underwent transurethral resection of the prostate (TURP) with a normal digital rectal examination, no history of prostate cancer and a PSA level of 2.5-10 ng/mL were studied. The total PSA (tPSA), percentage of free/total PSA (%fPSA), complexed PSA (cPSA), PSA density, cPSA density and the ratio of fPSA to cPSA were measured. Specificity, sensitivity, positive and negative predictive values were determined for all possible threshold values indicating the risk of incidental prostate cancer (T1a or T1b). Furthermore, the patients were subdivided according to age and the presence of an indwelling transurethral catheter. The areas under the receiver operating characteristic curves (AUC) were compared. RESULTS In the whole sample, the %fPSA was the best test predicting all incidental prostate cancers (AUC 0.618, reference: tPSA 0.494), whereas cPSA density was the best predictor of T1b disease (AUC 0.720, reference: tPSA 0.548). Stratification by age did not meaningfully alter the results, the presence of a transurethral catheter, however, was associated with a superiority of tests based on fPSA (AUC 0.620-0.670) compared with tests based on tPSA or cPSA (AUC 0.421-0.581). CONCLUSION Replacing tPSA by PSA derivatives (%fPSA or cPSA density) and stratifying by the presence of an indwelling transurethral catheter may improve the prediction of the risk of incidental prostate cancer and spare unnecessary biopsies before TURP in the tPSA range 2.5 10 ng/mL. PMID- 19191783 TI - A tertiary experience of urethral diverticulectomy: diagnosis, imaging and surgical outcomes. AB - OBJECTIVE To review the outcomes of consecutive patients referred with urethral diverticula to a tertiary centre; to investigate the diagnostic, imaging and surgical factors relevant to success. PATIENTS AND METHODS A retrospective case note review of 30 consecutive patients treated between January 1999-2007 was performed and data retrieved on demographics, presenting symptoms, preoperative imaging, surgical technique, outcomes and need for further intervention. RESULTS All patients were tertiary referrals, four after failed local repairs. The mean (range) interval between initial presentation and repair was 48 (1-264) months. Only seven patients (23%) had all three symptoms of the classical triad of dysuria, dyspareunia and dribble, whilst 23% did not have any of these symptoms. Transvaginal ultrasonography showed the diverticulum in six of nine patients, voiding cysto-urethrography (VCUG) in 13 of 18 patients (72%) and magnetic resonance imaging (MRI) in all 11 patients assessed. MRI accurately imaged diverticular configuration, whilst VCUG assessed detrusor and sphincteric function. Twenty-nine (97%) patients were cured of their diverticulum; all 19 patients with simple diverticula were cured at first attempt, whilst 17 procedures were performed on the 11 patients with complex diverticula. Twenty of 24 (83%) repairs were successful using three-layered closure, 9 of 11 using Martius interposition, and one using bulbospongiosus muscle interposition. There were three primary repair failures; two circumferential diverticula repaired with Martius interposition and one partial horseshoe diverticulum repaired without interposition had partial recurrences. Both were subsequently repaired successfully. One patient with chronic urethral pain from multiple, infected recurrences was eventually diverted. A pubovaginal sling procedure was required in only one (3.3%) patient with persistent pre-existing stress urinary incontinence (SUI). CONCLUSIONS The presentation of urethral diverticula is diverse and diagnosis frequently delayed. The most useful preoperative imaging is MRI and VCUG to assess diverticular anatomy and detrusor/urethral function, respectively. In simple cases, transvaginal excision with three-layer closure is curative, whilst more extensive, persistent or SUI-associated diverticula require Martius fat interposition. Sling procedures can be deferred until the results of primary excision are assessed. PMID- 19191784 TI - Can contemporary transrectal prostate biopsy accurately select candidates for hemi-ablative focal therapy of prostate cancer? AB - OBJECTIVE: To determine if biopsy characteristics can be used to identify men with unilateral prostate cancer on radical prostatectomy (RP) pathological specimens, thereby selecting candidates for hemi-ablative focal therapy. PATIENTS AND METHODS: Of 1458 men who had RP from January 2000 to June 2007, we identified 590 of 880 evaluable patients with unilateral disease on their preoperative biopsy. Charts were reviewed to record preoperative prostate-specific antigen (PSA) level, high-grade prostatic intraepithelial neoplasia (HGPIN), clinical stage, Gleason score, perineural invasion (PNI), prostate volume, number of positive cores, and percentage of positive cores. Final surgical pathology was evaluated for unilateral cancer. Univariate analysis was used (logistic regression method) to identify independent predictors of unilateral disease on the RP specimen. A subset analysis was done in men with low-risk disease, defined as clinical stage T1C, Gleason score <7 and a PSA level of <10 ng/mL. RESULTS: Of 590 men with unilateral disease on biopsy, 163 (27.3%) had unilateral disease on the RP specimen. Pathological features, including HGPIN (P = 0.714), Gleason score (P > 0.608), PNI (P = 0.714), number of positive cores (P = 0.076), percentage of cores positive (P = 0.056), prostate volume (P = 0.285), and PSA level (P = 0.062) did not improve the prediction of unilateral disease. When men with unilateral cancer were further stratified to include only those with low risk disease, 28.4% had unilateral disease on the RP specimen. None of the biopsy or clinical features evaluated were predictors of unilateral disease on the RP specimen. CONCLUSION: Unilateral prostate cancer on biopsy predicts unilateral disease on RP pathology in only 27.6% of cases. The predictive ability is not improved by adding biopsy and clinical characteristics. Additional methods are needed to accurately identify men appropriate for focal therapy. PMID- 19191785 TI - Regulation of growth factors in hormone- and drug-resistant prostate cancer cells by synergistic combination of docetaxel and octreotide. AB - OBJECTIVE To evaluate the effects of combined treatment with docetaxel and octreotide, a somatostatin analogue, on human hormone- and drug-refractory prostate cancer cell lines, PC-3 and DU-145, and on some growth factors related to tumour growth and angiogenesis in prostate cancer. MATERIALS AND METHODS A cell proliferation assay was used to assess the cytotoxicity of the drugs. To verify apoptosis, both DNA fragmentation (by enzyme-linked immunosorbent assay) and caspase 3/7 activity were measured. We also investigated the effect of combined docetaxel and octreotide on growth factors secreted from prostate cancer cells using a human growth factor antibody array. RESULTS The combination of docetaxel and octreotide resulted in significant synergistic cytotoxic activity and apoptosis, which was dose- and time-dependent. The combined treatment also resulted in significantly less secretion of stem cell factor and platelet-derived growth factor-AB in PC-3 cells, and transforming growth factor-beta and basic fibroblast growth factor in DU-145 cells, than in untreated controls. CONCLUSION Octreotide, a somatostatin analogue, combined with docetaxel might provide a rationale treatment option for hormone-refractory prostate cancer cells, not only by direct inhibition of cell proliferation but also by inhibiting the secretion of growth factors. PMID- 19191786 TI - Listeria monocytogenes infection in the face of innate immunity. AB - Pathogen recognition and induction of immune responses are important for efficient elimination of infection. However, pathogens such as Listeria monocytogenes employ strategies to evade or modulate these defences, thus creating a more favourable environment that ensures their survival and pathogenesis. New insights into these strategies, particularly those targeting innate immunity, have recently emerged. L. monocytogenes is initially detected at the cell surface or in phagosomes by toll-like receptor 2 and in the cytosol by nuclear oligodimerization domain (NOD)-like receptors (NOD1, NOD2) and NALP3 and Ipaf. It carries out N-deacetylation of peptidoglycan to avoid this detection by toll-like receptor 2 and NOD-like receptors. L. monocytogenes modulates transcription of host immunity genes through modification of histones and chromatin remodelling. Furthermore, L. monocytogenes has recently been shown to avoid autophagy and induce apoptosis in immune effector cells. In this review we discuss some of these strategies, which have provided new insights into the interaction between L. monocytogenes and the immune response at a crucial stage of infection. PMID- 19191787 TI - Listeria monocytogenes internalin and E-cadherin: from structure to pathogenesis. AB - Many bacterial pathogens that invade non-phagocytic cells first interact with host cell surface receptors. Adhesion to the host cell is followed by the activation of specific host signalling pathways that mediate bacterial internalization. The food-borne Gram-positive bacterium Listeria monocytogenes makes use of two surface proteins, internalin (InlA) and InlB to engage in a species-specific manner the adhesion molecule E-cadherin and the hepatocyte growth factor receptor Met, respectively, to induce its internalization. After entry, Listeria has the capacity to spread from cell to cell and disseminate to its target organs after breaching the intestinal, blood-brain and placental barriers in human. InlA but not InlB is critical for the crossing of the intestinal barrier, whereas the conjugated action of both InlA and InlB mediates the crossing of the placental barrier. Here we review the InlA-E-cadherin interaction, the signalling downstream of this interaction, the molecular mechanisms involved in bacterial internalization and the role of InlA-E-cadherin interaction in the breaching of host barriers and the progression to listeriosis. Together, this review illustrates how in vitro data were validated by epidemiological approaches and in vivo studies using both natural hosts and genetically engineered animal models, thereby elucidating key issues of listeriosis pathophysiology. PMID- 19191788 TI - Circulating Candida-specific anti-mannan antibodies precede invasive candidiasis in patients undergoing myelo-ablative chemotherapy. AB - The kinetics of circulating Candida mannan and anti-mannan antibodies were studied in consecutive plasma samples, obtained upon hospital admission, of 21 patients with microbiologically proven invasive candidiasis and 30 control patients who underwent myelo-ablative chemotherapy. The detection of Candida anti mannan antibodies preceded the diagnosis of invasive candidiasis in infected patients, and the antibodies were detected significantly more often in patients who had experienced multiple episodes of neutropenia than in the control group (OR 8.9, 95% CI 5.6-14.3; p <0.05). Mannan was predominantly detected in patients who developed invasive candidiasis during their first episode of neutropenia (OR 3.7, 95% CI 1.4-9.7; p <0.05). This observation suggests that patients with multiple episodes of neutropenia have been previously exposed to Candida and that the presence of anti-mannan antibodies in these patients might be associated with an increased risk of developing clinically manifest invasive candidiasis. PMID- 19191790 TI - Characterization of IS6110 insertions in the dnaA-dnaN intergenic region of Mycobacterium tuberculosis clinical isolates. AB - Mycobacterium tuberculosis isolates with identical IS6110 restriction fragment length polymorphism (RFLP) patterns are considered to be clonally related. The presence of IS6110 in the dnaA-dnaN intergenic region, one preferential locus for the integration of IS6110, was evaluated in 125 M. tuberculosis isolates. Five isolates had IS6110 inserted in this region, and two consisted of a mix of isogenic strains that putatively have evolved during a single infection. Strains from the same isolate had identical spoligo and mycobacterial interspersed repetitive unit-variable-number tandem repeat profiles, but had slight variations in IS6110 RFLP patterns, due to the presence of IS6110 in the dnaA-dnaN intergenic region. Duplication of the dnaA-dnaN intergenic region was found in one isogenic strain. PMID- 19191789 TI - Biofilm formation and prevalence of lukF-pv, seb, sec and tst genes among hospital- and community-acquired isolates of some international methicillin resistant Staphylococcus aureus lineages. AB - Methicillin-resistant Staphylococcus aureus (MRSA) is an important nosocomial agent of biopolymer-associated infections, and isolates of S. aureus can produce different virulence factors, including potent toxins. The biofilm formation and accumulation by certain international MRSA lineages were analysed, and the toxic shock syndrome-associated genes (tst, seb and sec) among these isolates were assessed. In addition, the presence of lukF-pv (encoding the F-subunit of Panton Valentine leukocidin (PVL)) was investigated. Most of the MRSA isolates tested were capable of forming biofilm on polystyrene surfaces, but lacked the superantigen toxin genes that were tested. PVL was rarely detected among the hospital isolates analysed. PMID- 19191791 TI - Early cortical bone healing around loaded titanium implants: a histological study in the rabbit. AB - OBJECTIVES: To identify the role of immediate implant loading on the early phases of the bone healing responses. MATERIAL AND METHODS: Implants were placed in rabbit tibial diaphyses and left to heal for 3, 7, 14, 28 or 42 days. Half of the animals received an immediate loading protocol of 2.2 N at 3 Hz for 1800 cycles and 5 days/week, whereas the others served as unloaded controls. Histological assessment was combined with histomorphometrical measurements. RESULTS: At early time-points, an endosteal and periosteal new bone formation was found, while the cortex itself contained damaged osteocytes. At later time-points, new bone formation was also found at the cortical level itself. Differences between groups were found mainly in this new bone formation process, with larger reactions for the endosteal and periosteal bone in the loaded group after 28 and 42 days, respectively. At the end-point of the experiment, bone formation at the cortical level was reduced in the loaded group compared with the control group. CONCLUSIONS: These results show that the immediate loading protocol caused no differences in the sequential events leading to osseointegration in cortical bone. However, the processes of new bone formation originating from the endosteum and the periosteum lasted longer compared with the unloaded controls. PMID- 19191792 TI - A feasibility study evaluating an in situ formed synthetic biodegradable membrane for guided bone regeneration in dogs. AB - PURPOSE: The aim was (1) to evaluate the soft-tissue reaction of a synthetic polyethylene glycol (PEG) hydrogel used as a barrier membrane for guided bone regeneration (GBR) compared with a collagen membrane and (2) to test whether or not the application of this in situ formed membrane will result in a similar amount of bone regeneration as the use of a collagen membrane. MATERIAL AND METHODS: Tooth extraction and preparation of osseous defects were performed in the mandibles of 11 beagle dogs. After 3 months, 44 cylindrical implants were placed within healed dehiscence-type bone defects resulting in approximately 6 mm exposed implant surface. The following four treatment modalities were randomly allocated: PEG+autogenous bone chips, PEG+hydroxyapatite (HA)/tricalcium phosphate (TCP) granules, bioresorbable collagen membrane+autogenous bone chips and autogenous bone chips without a membrane. After 2 and 6 months, six and five dogs were sacrificed, respectively. A semi-quantitative evaluation of the local tolerance and a histomorphometric analysis were performed. For statistical analysis, repeated measures analysis of variance (ANOVA) and subsequent pairwise Student's t-test were applied (P<0.05). RESULTS: No local adverse effects in association with the PEG compared with the collagen membrane was observed clinically and histologically at any time-point. Healing was uneventful and all implants were histologically integrated. Four out of 22 PEG membrane sites revealed a soft-tissue dehiscence after 1-2 weeks that subsequently healed uneventful. Histomorphometric measurement of the vertical bone gain showed after 2 months values between 31% and 45% and after 6 months between 31% and 38%. Bone to-implant contact (BIC) within the former defect area was similarly high in all groups ranging from 71% to 82% after 2 months and 49% to 91% after 6 months. However, with regard to all evaluated parameters, the PEG and the collagen membranes did not show any statistically significant difference compared with sites treated with autogenous bone without a membrane. CONCLUSION: The in situ forming synthetic membrane made of PEG was safely used in the present study, revealing no biologically significant abnormal soft-tissue reaction and demonstrated similar amounts of newly formed bone for defects treated with the PEG membrane compared with defects treated with a standard collagen membrane. PMID- 19191793 TI - A randomized, controlled clinical trial to evaluate a new membrane for guided bone regeneration around dental implants. AB - OBJECTIVES: The use of barrier membranes in guided bone regeneration (GBR) procedures for the treatment of alveolar bone defects is common practice. The objective of this study was to test whether a synthetic bioresorbable polyethylene glycol (PEG) hydrogel membrane could result in a similar amount of vertical bone fill as a standard collagen membrane, both combined with a membrane supporting material. MATERIAL AND METHODS: The study enrolled 37 patients requiring implant treatment with an expected osseous defect in the posterior maxilla or mandible. After raising a mucoperiosteal flap, the implant sites were prepared and dental implants placed. The defect height was then measured and defects <3 mm were excluded from the study. Defects were grafted with bovine bone mineral and randomly covered with either a collagen membrane (control group, 18 patients) or a PEG hydrogel membrane (test group, 19 patients), which is applied as a liquid. After a healing period of 6 months, surgical re-entry was performed and the change in vertical bone height from baseline evaluated. RESULTS: Well vascularized hard tissue was apparent at all sites and the regenerated bone was similar to the surrounding native bone. Mean vertical defect fill after 6 months was 5.63+/-1.84 mm at test sites and 4.25+/-1.16 mm at control sites, and the mean defect fills were 94.9% and 96.4% at test and control sites, respectively. More soft tissue complications were observed with the PEG membrane (e.g., delayed or incomplete wound healing) but all sites recovered uneventfully. CONCLUSIONS: The new PEG hydrogel membrane was as successful as a standard collagen membrane in the treatment of bony dehiscence defects around dental implants with simplified clinical handling. PMID- 19191794 TI - Long-term effects of magnetron-sputtered calcium phosphate coating on osseointegration of dental implants in non-human primates. AB - OBJECTIVES: To determine the effect of magnetron-sputtered calcium phosphate coating of implants on the later stages of osseointegration in a non-human primate model. MATERIAL AND METHODS: Eighteen and 20 implants with a 0.1 microm amorphous calcium phosphate coating and a turned surface, respectively, were inserted in the anterior upper and lower jaw of adult non-human primates. Following a 7.5 months healing period, one part of the implants remained in the submerged position. The other part of implants was connected to healing abutments to allow peri-implant inflammation to occur. After another 20 months, histologic and histomorphometric analysis of the peri-implant area was performed. RESULTS: Submerged implants with a calcium phosphate coating and a turned surface showed no signs of an inflammatory reaction. The histomorphometric parameters 'bone volume per tissue volume' (BV/TV) and 'bone-to-implant contacts' (BIC) were not affected by calcium phosphate coating. Non-submerged implants of both groups showed occasionally signs of inflammation at the implant-abutment junction. Histomorphometric analysis revealed that the distance between the implant abutment junction and the most coronal level (where bone was attached to the implant) as well as BV/TV and BIC were independent from the surface modification. CONCLUSION: Our results show that dental implants with calcium phosphate coating behave similar to turned implants independently whether they are connected to healing abutments or remain submerged. Ultra-thin calcium phosphate coating can combine the positive effects of calcium phosphate during the early stage of osseointegration without causing impairment of the later stages. PMID- 19191795 TI - Systemic tetracycline delays degradation of three different collagen membranes in rat calvaria. AB - OBJECTIVES: The aim of this study was to quantitatively evaluate the effect of systemic tetracycline (TTC) on the degradation of three different collagen membranes. MATERIALS AND METHODS: Collagen membranes were cut into 5 mm diameter membrane discs and labeled with aminohexanoyl-biotin-N-hydroxy-succinimide ester. One membrane disc each of a non-cross-linked [BioGide (BG)], glutaraldehyde cross linked [BioMend Extend (BM)], and ribose cross-linked [Ossix (OS)] was implanted on the calvaria of 40 Wistar rats. Another 10 biotinylated collagen membrane discs from each membrane type were processed for histologic observation and served as baseline; half of them (five from each group) were also treated with formic acid to inspect possible interference with biotinilazation of collagen by formic acid used during the decalcification process. A 10 mg/kg dose of TTC (50% of the minimal recommended antibacterial dose) to the experimental (20 animals) and saline to the control (20 animals) group was administered intramuscularly every 3 days. From each group, block sections were retrieved in half of the animals after 14 days and in the remaining after 28 days. Decalcified tissue histology was stained with streptavidin horseradish peroxidase. A computer assisted program measured the membranes' collagen contents. Statistical analysis consisted of analysis of variance (ANOVA) with repeated measures. RESULTS: No statistically significant differences in collagen contents were appreciated between biotinylated non-implanted membranes treated or not treated by formic acid. Systemic TTC had a different effect on the bio-degradation of the membranes: while it significantly decreased the resorption of two of the membranes (BG and BM), it had minimal influence on the ribose cross-linked membrane (OS). ANOVA with repeated measures, tests of within-subjects effects, showed a statistically significant difference between the membranes (P<0.001), within the membranes at the different time-points (P<0.001), a significant interaction between membranes and time and between the membranes and administered TTC (P<0.001). Test of between-subject effects revealed a statistically significant interaction with time and with TTC (P<0.001). CONCLUSIONS: Systemically administered TTC in sub-antibacterial doses may offer a possible treatment alternative to reduce bio-degradation and enhance bio-durability of certain collagen membranes. The findings of the present study could have clinical application in large non-self-contained bone defects, where prolonged membrane barrier functions are desirable. PMID- 19191796 TI - Effect of abutment's height and framework alloy on the load distribution of mandibular cantilevered implant-supported prosthesis. AB - OBJECTIVES: In cantilevered implant-supported complete prosthesis, the abutments' different heights represent different lever arms to which the abutments are subjected resulting in deformation of the components, which in turn transmit the load to the adjacent bone. The purpose of this in vitro study was to quantitatively assess the deformation of abutments of different heights in mandibular cantilevered implant-supported complete prosthesis. MATERIAL AND METHODS: A circular steel master cast with five perforations containing implant replicas (Theta 3.75 mm) was used. Two groups were formed according to the types of alloy of the framework (CoCr or PdAg). Three frameworks were made for each group to be tested with 4, 5.5 and 7 mm abutments. A 100 N load was applied at a point 15 mm distal to the center of the terminal implant. Readings of the deformations generated on the mesial and distal aspects of the abutments were obtained with the use of strain gauges. RESULTS: Deformation caused by tension and compression was observed in all specimens with the terminal abutment taking most of the load. An increase in deformation was observed in the terminal abutment as the height was increased. The use of an alloy of higher elastic modulus (CoCr) also caused the abutment deformation to increase. CONCLUSION: Abutment's height and framework alloy influence the deformation of abutments of mandibular cantilevered implant-supported prosthesis. PMID- 19191797 TI - Implant system for guiding a new layer of bone. Computed microtomography and histomorphometric analysis in the rabbit mandible. AB - OBJECTIVE: To prove the concept that an implant system with osteoconductive surface characteristics and an osteoinductive scaffold material has the capacity to guide vertical supracrestal bone growth in a rabbit mandible onlay model. MATERIAL AND METHODS: Thirteen adult white New Zealand rabbits each received custom-designed dental implants. All implants had sandblasted, acid-etched (SLA) surfaces, with the coronal aspect (3 mm) of each implant was left outside the lateral aspect of posterior mandibular bone, but covered by periosteum, muscle, subcutaneous tissue, and skin. Bone formation around implants placed adjacent to osteoinductive demineralized bone matrix (DBM) scaffolds were compared with contralateral implants without scaffolds in six rabbits using micro-CT imaging. Bone formation around implants with scaffolds from seven additional rabbits was measured using both micro-CT imaging and quantitative histology. RESULTS: At 8 weeks, new supracrestal bone was seen adjacent to all implants placed with DBM and two implants without DBM. The mean supracrestal bone heights achieved for implants with and without DBM scaffolds as measured by micro-CT was 2.1+/-0.9 and 0.8+/-0.9 mm, respectively (P=0.008). Histomorphometric analysis illustrated that supracrestal bone-to-implant contact for implants with DBM scaffolds was 58.1+/ 14% and that mean supracrestal bone height was 2.4+/-0.6 mm. CONCLUSIONS: Successful implant-guided supracrestal osteogenesis has been demonstrated in a rabbit model with the combined use of osteoconductive implant surfaces, an osteoinductive scaffold, and a device that prevents soft tissue downgrowth and provides scaffold stabilization. PMID- 19191798 TI - Analysis of the association of IL1B (C+3954T) and IL1RN (intron 2) polymorphisms with dental implant loss in a Brazilian population. AB - BACKGROUND: Although dental implants have a high success rate, failures occur, in spite of adequate clinical conditions. Together with the observation that multiple implant losses occur in certain groups of individuals (clusterization phenomenon), this suggests that host response may influence implant failure. Little is known about the influence of genetic susceptibility on implant loss. Interleukin (IL)-1 beta and IL-1 ra are believed to play a key role in the immune inflammatory response, and polymorphisms IL1B (C+3954T) and IL1RN (intron 2) are shown to alter the coding proteins expression. OBJECTIVES: The aim of this study was to investigate the association between dental implant loss and polymorphisms IL1B (+3954) and IL1RN (intron 2). MATERIAL AND METHODS: The study population (n=266) was divided into Test group (T)- 90 subjects with implant loss, and Control group (C)- 176 subjects without any implant failure. Genotyping was performed by PCR-RFLP. RESULTS: The number of present teeth was observed to influence implant loss. No differences in genotype and allele frequencies between C and T were found for IL1B (+3954) and IL1RN (intron 2) polymorphisms. However, the analysis of the whole study population (control and test groups) showed that genotype 2/2 was significantly more frequent in individuals with multiple implant losses (n=35) than in individuals that lost up to a single implant (n=231) (OR: 3.07, IC: 1.13-8.34, P=0.027). CONCLUSION: It was observed that number of teeth and edentulism were associated with implant loss. Genotype 2/2 of IL1RN polymorphism was significantly more frequent in patients who presented multiple losses, which suggests that the clusterization phenomenon has a genetic basis. PMID- 19191799 TI - Clinical experience with histidine-tryptophan-ketoglutarate solution in abdominal organ preservation: a review of recent literature. AB - Histidine-tryptophan-ketoglutarate (HTK) solution was introduced by Bretschneider as a cardioplegia solution in the early 1980s and has subsequently been applied to organ preservation for transplantation in Europe during the 1980s and in North America during the last six yr. With the increasing use of HTK for primary preservation, it is important that the transplant community be aware of the properties of this solution and the advantages and disadvantages of its use. Even if a center uses an alternative preservation solution, it is likely that import offers will be made for allografts that have been preserved in HTK. In this review article, recent literature describing the clinical use of HTK in abdominal transplantation will be summarized with references to earlier reports when indicated. PMID- 19191800 TI - Hematologic complications of anti-CMV therapy in solid organ transplant recipients. AB - Cytomegalovirus (CMV) infection complicates the post-operative course of patients receiving solid organ transplants. While ganciclovir has significantly reduced the direct effects of CMV infection, some patients cannot tolerate the optimal therapeutic exposure required for CMV prevention and treatment. Few reports directly address the incidence, consequences, and risk factors for hematologic toxicities related to ganciclovir therapy. Nevertheless, leukopenia, thrombocytopenia, and anemia occur in 5-50% of patients. Current strategies, focused on ganciclovir dose reduction, may increase the risk of CMV reactivation and drug-resistant disease. The current article reviews the incidence, risk factors, and consequences of ganciclovir-associated hematologic adverse events in transplant recipients. Management strategies, including ganciclovir dose reduction, and the addition of CMV hyperimmune globulin are discussed. Exposing this relatively frequently occurring, but uncommonly discussed, toxicity should lead to better avoidance and treatment strategies, without placing patients at increased risk of CMV disease. PMID- 19191801 TI - Dual hepatitis virus infections in liver transplant: case report and a review of the literature. AB - BACKGROUND: Liver transplantation (LT) using grafts from anti-HBVcore antibody positive (anti-HBVcAB+) donors carry risk for development of hepatitis B virus (HBV) infection. The long-term course of hepatitis C virus (HCV) patients receiving anti-HBVcAB+ grafts is poorly understood. PATIENTS AND METHODS: A patient with chronic hepatitis C received an anti-HBVc+ graft and developed de novo hepatitis B after four months. We describe the 14 HCV patients who received antiHBVc+ grafts and the condition of disease. RESULTS: Hepatitis B was treated successfully with lamivudine. One year later, breakthrough infection developed with a lamivudine-resistant mutant. Addition of adefovir led to HBV surface antigen to surface antibody seroconversion after two yr, which was maintained long term. Antiviral therapy was discontinued. Liver biopsy revealed minimal histologic changes up to eight yr post-LT. Survival of 14 recipients of antiHBVc+ allografts and 180 recipients of antiHBVc-negative grafts was equal (minimum follow up of five yr). Liver biopsies at four yr showed grade 0/1 and stage 0/1 in >70%; only two patients showed bridging fibrosis. A literature review of dual hepatitis virus infection revealed an overall milder course of hepatitis post-LT. CONCLUSION: The outcome of HCV patients receiving anti-HBc+ grafts is good and may be associated with a milder course of recurrent HCV. PMID- 19191802 TI - Post-transplant erythrocytosis: a disappearing phenomenon? AB - BACKGROUND: Erythrocytosis is relatively common post-kidney transplantation and may have adverse consequences. This study examined whether the incidence of erythrocytosis has remained stable over time and explored the impact of this condition on patient outcomes. METHODS: This was a retrospective single center review of an incidence cohort (transplanted between 1993 and 2005). Predictors of erythrocytosis and hemoglobin levels and subsequent patient and allograft survival were examined. RESULTS: Erythrocytosis (hemoglobin >170 g/L for >1 month) was observed in 59 of 511 recipients. Erythrocytosis developed in only 8.1% of those transplanted from 1997 to 2005, compared with 18.7% in those transplanted from 1993 to 1996 (p = 0.0005). Independent predictive factors were use of angiotensin converting enzyme inhibitors/angiotensin receptor blockers (ACEi/ARBs) (HR 0.176, 95% CI 0.040-0.71, p = 0.016), male gender (HR 3.72, 95% CI 1.54-9.0, p = 0.003), and mycophenolic acid agents (HR 0.49, 95% CI 0.237 0.99, p = 0.049). Patients with erythrocytosis had superior overall survival (HR for death 0.105, 95% CI 0.014-0.760, p = 0.026) but a trend for worse death censored graft loss (univariate HR 2.06, 95% CI 0.91-4.65, p = 0.084). CONCLUSIONS: The incidence of erythrocytosis is falling and is likely related to greater ACEi/ARB use and possibly more antiproliferative immunosuppression. Patient survival is excellent in those with erythrocytosis, but long-term graft survival may be compromised. PMID- 19191803 TI - Clinical significance of C4d deposition in stable renal allografts in the early post-transplantation period. AB - The clinical significance of C4d positivity in patients with stable graft function is undetermined. This study evaluated the clinical outcome of protocol biopsy-proven C4d-positive renal transplants with stable graft function in the early post-transplantation period. Protocol biopsies (n = 79) were performed on stable allografts on the 14th post-transplant day, and indication biopsies (n = 74) were performed on dysfunctioning allografts within one yr after transplantation. Clinical and histological findings, graft function and graft survival rates were compared between C4d-positive and C4d-negative grafts in each group. The incidence of C4d positivity was 5.1% in protocol biopsies and 9.5% in indication biopsies. In protocol biopsies, C4d-positive allografts showed minimal tubulointerstitial inflammation, and the graft function and graft survival rate did not differ from C4d-negative allografts. All C4d-positive allografts maintained stable graft function without anti-rejection therapy, and follow-up biopsies of two patients showed no C4d deposition or evidence of rejection. On the other hand, C4d-positive allografts in indication biopsies showed severe tissue injury, and the graft survival rate was significantly lower than C4d negative allografts. In conclusion, C4d-positive allografts with stable graft function in the early post-transplantation period take an indolent course. PMID- 19191804 TI - Fever of unknown origin in renal transplant patients with tacrolimus. AB - The immunosuppressive agent tacrolimus is now widely used for the prevention of acute and chronic rejection in renal allograft recipients. We here report on three patients, who developed drug-induced fever due to tacrolimus one to five months after renal transplantation. Extensive search for a focus, autoantibodies and virus infection remained inconclusive. Therefore, drug-induced fever was suggested. After discontinuing tacrolimus and switching to cyclosporine A fever completely resolved within 24 h. This report demonstrates that tacrolimus-induced drug fever should be included in the differential diagnosis of fever of unknown origin in renal transplant recipients. PMID- 19191805 TI - Orogenital ulcers in a liver transplant recipient: discerning between mycophenolate-mofetil-induced complication and Behcet's disease. PMID- 19191806 TI - Outcome predictors in African-American deceased-donor renal allograft recipients. AB - The relative importance of donor and recipient risk factors in predicting outcomes in African-American (AA) renal allograft recipients receiving contemporary immunosuppression, including early steroid withdrawal, has not been previously examined. We assessed the impact of 21 risk factors on five primary outcomes in 132 deceased-donor AA renal allograft recipients transplanted from July 2001 to August 2006 with follow-up 6-67 (mean 35 +/- 17) months by univariate and multivariate analysis. Thymoglobulin or basiliximab was given for induction, and mycophenolate mofetil with either tacrolimus or sirolimus (SRL) +/ prednisone for maintenance. Non-compliance accounted for 26% of graft loss (GL) and 19% of acute rejection (AR) episodes, and was more prevalent in patients who were HCV+ and those on prednisone. Delayed graft function remained a significant predictor of GL, but not via increased AR, and donor ethnicity emerged as an important predictor of patient death. De novo use of SRL resulted in increased AR, and only increased recipient age significantly predicted new-onset diabetes mellitus. Our preliminary results suggest the need for improvements in patient education, pre-transplant psychosocial assessment, and late post-transplant psychosocial support and can be utilized to help guide donor/recipient selection and tailor immunosuppressive management to optimize outcomes in this challenging group of patients. PMID- 19191807 TI - Longitudinal evaluation of mycophenolic acid pharmacokinetics in pediatric kidney transplant recipients. The role of post-transplant clinical and therapeutic variables. AB - This longitudinal study assessed the influence of post-transplant clinical and therapeutic variables in 50 kidney transplant recipients aged 2-19 yr receiving a triple immunosuppressive regimen consisting of cyclosporine microemulsion (CsA), steroids and MMF (300-400 mg/m(2) body surface area twice daily), the full pharmacokinetic profile (10 points) of which was investigated on post-transplant days 6, 30, 180 and 360. Total plasma MPA was measured by Enzyme Multiplied Immunoassay Technique. CsA therapeutic drug monitoring (TDM) was performed via C2 blood monitoring, while MPA TDM via C0. MPA Cmax, tmax, AUC0-12 and AUC0-4 pharmacokinetic profile changed significantly during the first post-transplant year. C0 was a poor predictor of the total MPA exposure [as measured by the area under the concentration-time curve AUC)], while a truncated AUC was a good surrogate of the 12-h profile (r = 0.91; p < 0.001) Graft function and cyclosporine therapy influenced MPA pharmacokinetics, as shown by the univariate and multivariate analyses. We conclude that because after transplantation MPA exposure varied over time, a strict TDM is advisable in the pediatric population. PMID- 19191808 TI - Coverage of the organ donation process on Grey's Anatomy: the story of Denny Duquette. AB - This investigation examined the impact of Grey's Anatomy viewing on a host of organ donation-related variables: organ donation beliefs and attitudes and a willingness to discuss organ donation with family. Following exposure to a storyline in which two myths were presented: (i) the rich and famous can buy their way to the top of organ waiting lists (purchase myth) and (ii) friends and family of medical professionals receive organ transplants quicker than other individuals (relationship myth), results revealed that loyal viewers of Grey's Anatomy are less likely to believe the purchase myth compared to non-viewers. No difference emerged between these two groups with respect to the relationship myth. Additionally, loyal viewers were more likely to talk about their willingness to donate organs than non-viewers. Results are discussed with an emphasis on how Grey's Anatomy portrayals can impact perceptions and action tendencies related to organ donation. PMID- 19191810 TI - Assessment of liver graft function and regeneration by galactosyl-human serum albumin (99mTc-GSA) liver scintigraphy in adult living-donor liver transplantation. AB - BACKGROUND: In adult living-donor liver transplantation (LDLT), the assessment of the graft functional reserve is very important. We evaluated the graft functional reserve by technetium-99m-diethylenetriaminepenta-acetic acid-galactosyl-human serum albumin ((99m)Tc-GSA) liver scintigraphy. PATIENTS AND METHOD: From May 2003 to September 2006, (99m)Tc-GSA studies were performed in 27 adult recipients on two, four wk after LDLT, the receptor index [ratio of liver to heart-plus liver radioactivity at 15 minutes (LHL15)] (LHL15) was calculated. Recipients were divided into two groups according to LHL15 on two wk after LDLT (group H; >0.935, group L; <0.935). Liver functional tests and recipients' background parameters were evaluated between the two groups. RESULT: Group L accompanied higher preoperative model for end-stage liver disease (MELD) score (p = 0.038), lower graft-recipient weight ratio (GRWR) (p = 0.032) and older donor age (p = 0.003) compared with group H. There was no significant difference in the graft regeneration rate between two groups. The three-yr cumulative survival rate was 76.1% in group L and 88.9% in group H. CONCLUSION: In LDLT, LHL15 has the potential to assess the graft function and predict the recipients' outcome. Graft function after LDLT may be related closely to the pretransplant MELD score, GRWR, and donor age. PMID- 19191809 TI - Domino liver transplantation using a graft from a donor with familial hypercholesterolemia: seven-yr follow-up. AB - A 46-yr-old female with hepatocellular carcinoma and severe hepatitis B-related liver cirrhosis received a domino liver graft from a 25-yr-old female with homozygous familial hypercholesterolemia (HFHC) in September 2001. Hypercholesterolemia occurred in the graft recipient within one yr after transplantation and was partially controlled by atorvastatin. Three yr after transplantation, an autologous CD34(+) cell transplantation was performed in order to better control the hypercholesterolemia. Only preliminary results of this domino liver transplantation (DLT) were published in 2003, without a long term analysis of the hypercholesterolemic effects in recipient. Subsequent to DLT, the average plasma cholesterol level in the domino donor rapidly normalized and seven yr after had a value of 182 mg/dL. After seven-yr follow-up, the domino recipient has no hepatocarcinoma recurrence. Moreover, no signs of cardiovascular or atherosclerotic lesions were noted despite an elevated plasma cholesterol level (339 mg/dL after seven yr of follow-up) resistant to drug therapy and stem cell autotransplantation. In conclusion, DLT using a liver graft from a patient with HFHC provides a viable option for marginal recipients. PMID- 19191811 TI - The influence of chronic periodontitis on serum TNF-alpha, IL-6 and hs-CRP concentrations, and function of graft and survival of kidney transplant recipients. AB - OBJECTIVE: The aim of the study is to analyze whether chronic periodontitis (CP) influences serum tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, and high sensitivity C-reactive protein (hs-CRP) concentrations in renal transplant recipients and patients or graft survival. BACKGROUND: CP is associated both with higher serum concentrations of cytokines and CRP and high cardiovascular risk. METHODS: One hundred and seventeen patients with a transplanted kidney (Tx) were divided according to the Community Periodontal Index of Treatment Needs (CPITN) into two subgroups: TxP+: severe CP (CPITN 3-4) and TxP-: no or moderate CP (CPITN 0-2). The control group (C) consisted of 36 subjects with no kidney disease and with no or moderate CP (CPITN 0-2). RESULTS: hs-CRP concentration was higher in TxP+ than in TxP- patients [2.0 mg/L (0.7-4.4) vs. 0.9 mg/L (0.3-1.9), p = 0.006]. There were no differences in serum TNF-alpha and IL-6 between TxP+ and TxP-. Significant positive correlation between CPITN score and hs-CRP concentration was found both in Tx and C. The hazard ratio of death was 7.17 (1.4 76.4) for TxP+ patients. CP status did not increase the risk of graft loss or doubling of serum creatinine. CONCLUSIONS: Severe CP is associated with increased serum hs-CRP concentration in patients after kidney transplantation. Severe periodontitis seems to increase the risk of patients' death after kidney transplantation. PMID- 19191812 TI - The impact of meticulous management for hepatic artery thrombosis on long-term outcome after pediatric living donor liver transplantation. AB - To analyze the risk factors in the development of hepatic artery thrombosis (HAT) and assess the impact of our perioperative management for HAT on the long-term outcome after pediatric living donor liver transplantation (LDLT), we reviewed 382 patients under 12 yr of age who underwent 403 LDLT from January 1996 to December 2005. One- and 10-yr patient survival rates were 78% and 78% in the patients with HAT (27 patients; 6.7%), and 84% and 76% in the patients without HAT, respectively (p = n.s.). Univariate analysis showed gender (female), body weight (lower), and graft-to-recipient weight ratio (higher) were significant risk factors in the patients with HAT (p < 0.05). Patients with Doppler ultrasound signal loss of the hepatic artery (HA) accompanied by an increase of liver enzymes underwent thrombectomy and reanastomosis (S-group, n = 13), and patients with a weak HA signal underwent anticoagulant therapy (M-group, n = 13). One patient underwent re-LDLT. One- and five-yr patient survival rates were 83% and 83% in the S-group, and 77% and 77% in the M-group (p = n.s.). The incidence of biliary complications in the S-group (58%) was significantly higher than that of the M-group (15%). For a successful long-term outcome, the early detection of HAT and prompt medical and surgical intervention are crucial to minimize the insult of HAT. PMID- 19191813 TI - Outcome of hepaticojejunostomy for biliary tract obstruction following liver transplantation. AB - BACKGROUND: Strictures and concrements are the most common biliary complications following liver transplantation. Endoscopic treatment might not lead to a definitive cure in all patients, especially in strictures involving the biliary bifurcation. The aim of this study was to determine the efficacy and the long term outcome of hepaticojejunostomy (HJS) for post-transplant biliary tract obstruction. MATERIAL AND METHODS: Thirty-seven patients were retrospectively studied for resolving of cholestasis and the incidence of recurring biliary obstruction. RESULTS: Surgery was performed because of anastomotic strictures in 11, ischemic strictures at the donor common bile duct in seven, strictures involving the bile duct bifurcation in 10, hepatolithiasis without strictures in one and biliary cast formation diagnosed by endoscopic retrograde cholangiography or T-tube cholangiography in eight patients. Cholestasis instantly improved in 82% of the patients. After a long-term follow-up of median 33 months (range 3 149), 28 of the patients (76%) required no further intervention for recurring biliary obstruction following HJS. Anastomotic strictures were observed in six (16%), recurring biliary concrements in two patients (5%). CONCLUSION: HJS did prevent recurrent biliary obstruction in the majority of the patients. We therefore recommend early HJS for complicated post-transplant biliary tract obstruction not treatable by a limited number of endoscopic interventions. PMID- 19191814 TI - From words to action--influence of two organ donation campaigns on knowledge and formal decision making. AB - INTRODUCTION: In Sweden, two donation campaigns have been carried out; one short term (STC) during October 2001, and one long term (LTC) between the years 2003 2005. The goal was not only to inform the public but also to create a positive attitude, make people talk about donation and formally declare their decision. METHODS: The effects of the two campaigns were evaluated through three opinion polls. RESULTS: The willingness to donate was widespread (86%) before and after the campaigns. The LTC increased the knowledge of the Donor Card (24-35%, p < 0.001) and the National Donor Registry (19-40%, p < 0.001). The LTC focused on making people register, still though, only a small increase of registrants was found (11-14%, p = 0.043). The proportion expecting the question of organ donation if their relative was to die under circumstances that made organ donation possible increased (73-79%, p = 0.002). No corresponding increase was found regarding the number having informed a relative about their decision (20 23%, ns). CONCLUSION: The LTC was successful in increasing the expectation for the donation request. It also improved the knowledge of the Card and the Registry and slightly increased the number having registered. However, neither of the campaigns succeeded in making people inform their relatives. PMID- 19191816 TI - Roux limb volvulus after pancreas transplantation: an unusual cause of pancreatic graft loss. AB - Pancreas transplantation with enteric drainage avoids the long-term urological complications of bladder drainage. Increasing use of this technique raises the possibility of complications from the enteric reconstruction. This report describes a patient five yr after left-sided pancreas transplant with Roux-en-Y enteric drainage, presenting with abdominal pain, leukocytosis and radiological evidence of bowel obstruction. Exploration revealed a volvulus of the Roux limb as it passed through the mesocolon, with necrosis of the allograft duodenum and marked congestion of the pancreas. This is the first report of pancreas graft loss due to this entity, which should be recognized as an unusual cause of abdominal pain after pancreas transplantation. Potential bowel complications related to the sigmoid mesentery in left-sided pancreas transplantation are additional reasons for right-sided placement of the pancreas allograft. PMID- 19191815 TI - Transplant tourism to China: the impact on domestic patient-care decisions. AB - Organ procurement in China has been criticized because of its reliance on executed prisoners as donors. We aimed to assess the influence of perceptions about organ procurement practices in China on domestic patient-care decisions. METHODS: An anonymous internet administered case-based questionnaire was used to survey a sample of healthcare professionals with affiliations to hepatology and transplantation professional societies. RESULTS: Of 674 completed surveys, the vast majority (93%) of the respondents were physicians, surgeons or allied transplant professionals actively caring for liver transplant patients and 81% practiced in the US. A strong majority believed procurement practices were ethically sound in the US and Europe (87% and 73%) but fare fewer believed that procurement practices were ethically sound in China (4%, p < 0.001). In case based questions, lack of confidence in the ethical standards of organ procurement in China predicted patient-care decisions. The majority would provide post transplantation care for patients who underwent liver transplantation at another domestic center, in a foreign country and in China (90%, 78%, and 63%, respectively, p < 0.001) yet respondents who suspected unethical procurement practices in China were more reluctant to do so (p < 0.001). CONCLUSIONS: Transplant professionals expressed concern about organ procurement practices in China which influenced their patient-care decision-making. PMID- 19191817 TI - Two cases of central venous catheter-related thrombosis in living liver donors: how can the risk be minimized? AB - A central venous catheter (CVC) is commonly used for intraoperative management by anesthetists and surgeons during major operations, including donor operations for living donor liver transplantation (LDLT), in which donor safety is of utmost importance. Reasons for use of CVC for donors include measurement of central venous pressure and drug infusion when necessary. A potentially serious complication of a major operation is pulmonary thromboembolism. We report two cases of LDLT donors complicated by catheter related thrombosis (CRT) of the jugular vein, who were eventually discharged without long-term complications. To the best of our knowledge, there has been no report of CRT among LDLT donor population. In this report, in order to minimize the risks related to CRT in LDLT donors, we propose thorough screening for thrombophilic disorders, use of a silicone or polyurethane double-lumen CVC as thin as possible, placement of the tip of the CVC at the superior vena cava via the right jugular vein using ultrasonography as a guide for puncture, and removal of the catheter at the end of the operation based on our experience of CRT among LDLT donors. PMID- 19191818 TI - Predictors of untreated dental decay among 15-34-year-old Australians. AB - OBJECTIVES: To determine predictors of untreated dental decay among 15-34-year olds in Australia. METHODS: Data were from Australia's National Survey of Adult Oral Health, a representative survey that utilized a three-stage, stratified clustered sampling design. Models representing demographic, socioeconomic, dental service utilization and oral health perception variables were tested using multivariable logistic regression to produce odds ratios. RESULTS: An estimated 25.8% (95% CI 22.4-29.5) of 15-34-year-old Australians had untreated dental decay. After controlling for other covariates, those who lived in a location other than a capital city had 2.0 times the odds of having untreated dental decay than their capital city-dwelling counterparts (95% CI 1.29-3.06). Similarly, those whose highest level of education was not a university degree had 2.1 times the odds of experiencing untreated dental decay (95% CI 1.35-3.31). Perceived need of extractions or restorations predicted untreated coronal decay, with 2.9 times the odds for those who perceived a treatment need over those with no such treatment need perception (95% CI 1.84-4.53). Participants who experienced dental fear had 2.2 times the odds of having untreated dental decay (95% CI 1.38-3.41), while those who reported experiencing toothache, orofacial pain or food avoidance in the last 12 months had 1.9 times the odds of having untreated dental decay than their counterparts with no such oral health-related quality-of-life impact (95% CI 1.20-2.92). The multivariate model achieved a 'useful' level of accuracy in predicting untreated decay (area under the ROC curve = 0.74; sensitivity = 0.63; specificity = 0.73). CONCLUSIONS: In the Australian young adult population, residential location, education level, perceived need for dental care, dental fear, toothache, orofacial pain or food avoidance together were predictors of untreated dental decay. The prediction model had acceptable specificity, indicating that it may be useful as part of a triage system for health departments wishing to screen by means of a questionnaire for apparently-dentally healthy 15-34-year-olds. PMID- 19191819 TI - Barriers to restorative care as perceived by dental patients attending government hospitals in Tanzania. AB - OBJECTIVES: Perceptions of dental patients in Tanzania regarding barriers to restorative care were examined. METHODS: A total of 1138 dental patients aged 15 years and above, attending 12 selected government dental clinics in January 2005, completed a pre-tested questionnaire. anova and logistic regression were used to identify the effects of independent variables upon the barrier factors. RESULTS: Ten barriers to restorative care in Tanzania were identified. The five main barriers included: No advice received from dentist (mean = 3.82); Lack of knowledge on restorative care (mean = 3.58); Lack of habits for going for dental check-ups (mean = 3.31); Bother that one may get in seeking restorative care (mean = 3.28) and Past experience with dental treatment (mean = 3.25). Less respondents in the middle economic zone perceived issues related to beliefs and misinformation, lack of knowledge on restorative care and past experience with dental treatment to be barriers than respondents in low and high economic zones (P < 0.01). However, more of them perceived issues related to high fee for restoration, bother that one may get in seeking restorative care and few clinics rendering restorative care (P < 0.0001) to be barriers than respondents in the high economic zone did. Women, more than men, perceived few clinics rendering restorative care and no advice received from dentist as barriers to restorative care (P < 0.01). Respondents who had received a restoration and/or had relatives who had received a restoration perceived bother that one may get in seeking restorative care and no advice received from dentist more as barriers to restorative care than did those who lacked that experience and whose relatives had not received a restoration (P < 0.05). CONCLUSIONS: No advice received from dentist and lack of knowledge on restorative care, were the two major barriers to restorative care, as perceived by dental patients in Tanzania. Implementation of the Atraumatic Restorative Treatment approach may overcome many of the barriers identified. PMID- 19191820 TI - School-level contextual factors associated with betel quid chewing among schoolchildren in Taiwan. AB - OBJECTIVES: Previous to this research, no literature had explored the clustering of betel quid chewing at the school-level and the contextual factors contributing to this behavior among elementary schoolchildren. This study therefore examines this in clusters stratified according to school geography and the rate of aboriginal students in each school. Individual-level characteristics are controlled in the study. METHODS: The data used derive from student and parent surveys of the School Smoking Survey Project from a sampling of third and fourth grade children from 13 elementary schools in Tao-Yuan County, Taiwan (n = 1585). The individual- and school-level characteristics regarding betel quid chewing were estimated using multi-level logistic regression analysis. RESULTS: Compared to their counterparts, students attending schools with higher percentages of aboriginal students (OR = 3.60, 95% CI: 1.31-9.95), or those schools located in a mountainous region (OR = 2.82, 95% CI: 1.19-6.70), were found to have a greater risk of chewing behavior. Other observed characteristics having a significant relationship to student chewing behavior were drinking alcohol and smoking cigarettes. CONCLUSIONS: The new findings from this study suggest that the specific school attended is an important point of intervention. Therefore, effective policies and preventive programs should be strongly considered for implementation in schools that are putting students at the greatest risk for betel quid chewing behavior. Our study also confirmed the relationship between betel quid chewing, cigarette smoking, and alcohol drinking, suggesting that multifaceted intervention strategies need to be considered in regard to such behaviors among elementary schoolchildren. PMID- 19191821 TI - Is there equity in oral healthcare utilization: experience after achieving Universal Coverage. AB - OBJECTIVES: To assess the socioeconomic-related (in)equality and horizontal (in)equity in oral healthcare utilization among Thai adults after Universal Coverage (UC) policy implemented nationwide, and to decompose the source of inequality in utilization. Further, to identify the determinants that effect to out-of-pocket payments for oral healthcare. METHODS: Using the data of 32748, Thai adults aged 15 years and over from nationally representative Health and Welfare Survey and Socio-Economic Survey in 2006. This study employs concentration index (CI) and horizontal inequity index (HI) to measure the socioeconomic-related inequality and horizontal inequity in oral healthcare utilization, respectively. Further, employing decomposition method to identify the sources of inequality comprising of a contribution of income, need determinants (i.e. self-assessed oral health, demographic characteristics), non need determinants (i.e.working status, educational level attainment, type of insurance entitlement, geographic characteristics and marital status) and residual term. Two-part model is used to determine the factors effect to out-of pocket payments for oral healthcare. RESULTS: There are the pro-rich inequality and inequity in oral healthcare utilization among Thais as indicated by significantly positive values of CI (=0.199) and HI (=0.206). The poor are more likely to access and utilize services at subsidized public facility particularly community hospital, as opposed to the better-off who tend to utilize services at private facility. Income and non-need determinants principally contribute to the pro-poor in public sector utilization, unlike pro-rich in private sector utilization. Need factors account for most of the pro-poor utilization. Type of treatment obtained and insurance used in the last visit are the substantial determinants effect to incurring out-of-pocket payments for oral healthcare. CONCLUSION: Notwithstanding the UC policy implementation made impressive strides toward improving of welfare coverage and an increase in accessibility of health services among Thais, inequality and inequity in oral healthcare utilization persist even when the country achieved universal coverage. Decomposition analyses demonstrate the association of each determinant to inequality in utilization which provides information for policy amendment to achieve the goal of equity in healthcare system. PMID- 19191822 TI - Prevalence and factors associated to dental trauma in infants 1-3 years of age. AB - The aims of this study were to assess the epidemiology of traumatic dental injuries (TDI) to primary teeth in infants and toddlers between 1 and 3 years of age and investigate whether TDI was related to biological and social factors. A representative sample of infants and toddlers in the city of Belo Horizonte, Minas Gerais, Brazil (n = 519) was examined during a vaccination campaign. The evaluation of clinical signs of previous dental trauma was performed by nine previously calibrated examiners (intra-examiner Kappa = 0.95%; inter-examiner Kappa = 0.88%). Data were analysed through descriptive analysis, the chi-squared test (P < 0.050) and logistic regression. The prevalence of dental trauma was 41.6%. Among the 519 children examined, 193 (37.2%) had enamel fractures, 30 (5.7%) had enamel-dentin fractures and only three (0.6%) had enamel-dentin fractures involving the pulp. Dental care had been performed within the first 24 h in 4.1% of the sample and after 36 h in 0.8% of the sample. There was no statistically significant difference between genders. The most reported aetiologies were falls (28.8%) and collisions (6.8%). There were no statistically significant associations between the prevalence of dental trauma and non nutritive sucking habits, dental caries or lip incompetence (P > 0.050). Children of mothers with low levels of schooling (0-6 years of study) had a greater prevalence of TDI (P = 0.001). The results of the logistic regression also demonstrated a significant association between dental injury and the Social Vulnerability Index (P = 0.045). Children from families with high social vulnerability (worse living conditions) had a 1.51 (95% confidence interval 1.0 2.2) greater chance of exhibiting TDI. These results can help guide the implementation of health promotion policies. PMID- 19191823 TI - Root formation of an autotransplanted tooth. AB - Traumatic injuries with loss of anterior teeth occur more frequently in children. Treatment of the dental trauma is complex and requires a comprehensive and accurate diagnostic and treatment plan. It is also important to consider the biological, functional, esthetic, and economic aspects. The purpose of this article is to show a case report with a successful root formation and periodontal healing after autotransplantation of maxillary second right premolar in place of maxillary right central incisor site. Tooth autotransplantation may be considered as a viable treatment option instead of a conventional prosthetic and implant rehabilitation. PMID- 19191824 TI - Autotransplantation of first premolar to replace a maxillary incisor - 3D-volume tomography for evaluation of the periodontal space. AB - The anterior maxilla is the most traumatized region during childhood and tooth loss is frequently involved. Space closure with implants is contraindicated in growing patients and maintenance of space is necessary over a long period. Transplantation of premolars from the mandible with two-third to three-quarter root formation wide open foramen provides the best prognosis for permanent survival. CASE REPORT: This report describes the management of a 10 year old girl with autotransplantation to replace an upper incisor by a lower premolar under consideration of special care for the periodontal ligament during surgical procedure. A 3-D imaging with digital volume tomography (DVT) was used 20 months after transplantation to evaluate the periodontal morphology. DISCUSSION: Autotransplantation of premolars with incomplete root formation to replace maxillary incisor is a treatment alternative after traumatic loss of teeth. There were no signs of pathosis like resorption or ankylosis and a normal periodontal space demonstrated with DVT. There is no need for endodontic treatment before transplantation. In this case of autotransplantation of a premolar, the DVT after 20 months gives evidence of a successful regeneration of the periodontal ligament. PMID- 19191825 TI - Maternal genitourinary infection and risk of cerebral palsy. AB - AIM: To examine the association between genitourinary infection during pregnancy and cerebral palsy (CP) in children. METHOD: Medicaid and birth certificate data were obtained for 135 835 pregnant women with singleton births paid for by Medicaid from 1996 to 2002. Linked Medicaid billing data were obtained for their children in 2007. The association between maternal genitourinary infection and CP was modeled using generalized estimating equations. RESULTS: Maternal genitourinary infection was significantly associated with CP (odds ratio [OR]=1.27, p=0.007). Additional analyses revealed that the association was strongly significant for preterm or low birthweight infants when maternal infection was diagnosed in the first two trimesters of pregnancy (OR=1.62, p<0.001). This association remained (OR=1.72, p<0.001) when the model was limited to cases of CP diagnosed by at least two different clinicians. Infection was not significantly associated with CP in term or normal-birthweight infants. INTERPRETATION: Maternal genitourinary infection occurring in the first two trimesters was associated with increased risk of CP in preterm or low-birthweight children. Additional research is needed to determine whether this association is affected by antimicrobial treatment. PMID- 19191826 TI - Cerebrospinal fluid neopterin in paediatric neurology: a marker of active central nervous system inflammation. AB - AIM: Cerebrospinal fluid (CSF) neopterin production is increased by interferon gamma stimulation and appears to act as a marker of intrathecal immune activation. We aimed to test the usefulness of elevated CSF neopterin as a biological marker of central nervous system (CNS) inflammation. METHOD: We retrospectively reviewed CSF neopterin in 158 children (89 males, 69 females, mean age 4y 1mo, SD 3y 11mo, range 1mo-15y). RESULTS: CSF neopterin levels in children with chronic static CNS disorders (n=105) were predominantly low, suggesting that inflammation is rare in these patients. We created an upper value of normal (chronic static group 95th centile 27.4 nmol/l). CSF neopterin was elevated in all 10 patients with acute encephalitis and in 10 of 12 patients with other acute inflammatory CNS disorders (demyelination, post-infectious ataxia, myelitis). CSF neopterin was also significantly elevated in patients with chronic progressive disorders of inflammatory origin. Interestingly, CSF neopterin was elevated in four of six patients with chronic static disorders who were tested during a febrile exacerbation of seizures or dystonia, suggesting that intrathecal immune activation may be important in this setting. INTERPRETATION: Neopterin has a short half-life and was useful for monitoring inflammation activity in a patient with relapsing-remitting encephalitis. CSF neopterin is a useful marker of inflammation in a broad range of acute and chronic CNS disorders, and is a significantly more sensitive marker of inflammation than CSF pleocytosis. PMID- 19191827 TI - Neurodevelopmental outcomes in children with cerebellar malformations: a systematic review. AB - Cerebellar malformations are increasingly diagnosed in the fetal period. Consequently, their consideration requires stressful and often critical decisions from both clinicians and families. This has resulted in an emergent need to understand better the impact of these early life lesions on child development. We performed a comprehensive literature search of studies describing neurodevelopmental outcomes of cerebellar malformations between January 1997 and December 2007. Overall, the data suggested that children with isolated inferior vermis hypoplasia (IVH) and mega cisterna magna (MCM) have a good developmental outcome, whereas children with molar tooth sign/Joubert syndrome, vermis hypoplasia, pontocerebellar hypoplasia (PCH) type II, and cerebellar agenesis experience moderate to severe global developmental delays. Reports for Dandy Walker malformation (DWM) were conflicting; however, the presence of a normally lobulated vermis and the absence of associated brain anomalies were associated with a more favourable outcome. Finally, children with isolated cerebellar hypoplasia experienced fewer impairments. Important methodological limitations highlighted include a lack of standardized outcome measure use in 79% of studies and the predominant use of retrospective study designs (85%), with 40% limited to case reports or case-series. In summary, rigorous outcome studies describing the spectrum of disabilities in survivors are urgently needed to accurately delineate the long-term neurodevelopmental consequences of cerebellar malformations. PMID- 19191828 TI - Limbic encephalitis associated with anti-GAD antibody and common variable immune deficiency. AB - A variety of autoantibodies have been identified with complex neurological disorders including limbic encephalitis. The underlying trigger for the immune mediated process and the role of autoantibodies in the pathogenesis of limbic encephalitis remain to be clarified. Here, we report a 16-year-old female who was diagnosed with acute-onset non-neoplastic limbic encephalitis. The initial treatment with pulse doses of i.v. methylprednisolone improved the neurological symptoms. During the next 12 months, progressive decline was reported in her academic functioning and seizure control. Additional diagnostic evaluation revealed no evidence of malignancy or central nervous system infection but circulating anti-GAD antibodies were present in the serum and cerebrospinal fluid. Intravenous gammaglobulin infusion was initiated and continued monthly. Intravenous and oral steroids were added to the intravenous immunoglobulin treatment because of the worsening course and seizures, despite treatment with antiepileptic medications. Screening for quantitative immunoglobulins demonstrated hypogammaglobulinaemia with low immunoglobulin M and G in addition to low immunoglobulin A levels. There was a lack of protective pneumococcal antibody titers before and after immunization. Therefore, common variable immunodeficiency was suspected despite there being no history of recurrent infections. To our knowledge, this is the first report describing a possible link between immune-mediated limbic encephalitis and immune deficiency. PMID- 19191829 TI - The ketogenic diet improves recently worsened focal epilepsy. AB - AIM: We observed a dramatic response to the ketogenic diet in several patients with highly refractory epilepsy whose seizure frequency had recently worsened. This study aimed to identify whether this characteristic was a useful indication for the ketogenic diet. METHOD: From the 70 patients who received the ketogenic diet during a 3-year period at our institution, we retrospectively selected patients with focal epilepsy. There were 22 children, 13 females and nine males, aged from 5 months to 18 years 6 months (mean 6y 9mo, SD 5y 11mo). Fifteen had symptomatic and seven had cryptogenic focal epilepsy. Seizure frequency 1 week before initiating the ketogenic diet was compared with that at 1 month and at the last visit on the diet. RESULTS: Eleven patients were responders (defined as reduction of seizures by more than 50%) at 1 month. Responders were higher (p=0.046) in the group with a recent worsening of seizures than in those with stable seizure frequency. Seven patients were still seizure-free at 6 months on the diet. Tolerability was excellent in 10 patients. Five patients stopped the diet because of early side effects. INTERPRETATION: The ketogenic diet may be a valuable therapeutic option for children with pharmacoresistant focal epilepsy, particularly those with a recent deterioration of seizure control and neurological status. Because of its rapid effect, the ketogenic diet may be a useful support to intravenous emergency drugs in such a situation. PMID- 19191832 TI - Are infants with torticollis at risk of a delay in early motor milestones compared with a control group of healthy infants? AB - Recently it has been claimed that infants with congenital muscular torticollis (CMT) are at risk of a delay in early motor milestones. The aim of the present study was to investigate whether infants with CMT are indeed at risk in comparison with a control group of healthy infants. A second aim was to investigate whether the time spent in a prone position and plagiocephaly had any influence on motor development. Eighty-two infants with CMT (35 females and 47 males) were compared with 40 healthy infants (18 females and 22 males). Motor development was assessed with the Alberta Infant Motor scale (AIMS). Multiple regression showed that infants in the CMT group had a significantly lower AIMS score than the control group at 2 months (p=0.03) and 6 months of age (p=0.05). Infants who spent at least three occasions daily in a prone position when awake had significantly higher AIMS scores than infants who spent less time prone at 2 months (p=0.001), 6 months (p<0.001), and 10 months of age (p<0.001). The CMT group achieved early motor milestones significantly later than the control group until the age of 10 months, but the risk of delay seems to be more strongly associated with little or no time prone when awake than with CMT. PMID- 19191833 TI - Quality of life of adolescents with cerebral palsy: perspectives of adolescents and parents. AB - Quality of life (QOL) has emerged over the past 20 years as an outcome for measuring the effectiveness of health-improvement interventions. The Cerebral Palsy Quality of Life Questionnaire for Children (CPQOL-Child) is well regarded and now integrated into research internationally. We describe the results of qualitative research, using grounded research in which we aimed to identify domains of QOL for adolescents with cerebral palsy (CP) from adolescent and parent perspectives to guide the development of an adolescent version. Seventeen adolescents (nine males, eight females) aged 13 to 18 years (mean=15.53 SD=1.74), with varying levels of impairment (Gross Motor Function Classification System levels I n=5, II n=2, III n=6, IV n=4, and V n=6) and their parents (n=23) participated in semi-structured interviews. Questions included: 'What do you think is important to your (child's) QOL?' and 'How does CP impact on your (child's) life?' Fifteen themes were identified, including domains related to health issues in adolescence, participation, education, specific CP-related issues (pain and discomfort, communication), family issues, practical issues (financial resources), and changes associated with adolescence (sexuality, independence). The composition of these QOL domains reflects the developmental concerns of adolescents with CP, adding to the views of children, and strongly supports adolescent participation in the development of self-reported well-being and QOL measures. Many of the domains are consistent with child reports of QOL and thus it appears feasible and valid to develop a measure that will be transferable across childhood and adolescence. PMID- 19191834 TI - Interactive computer play in rehabilitation of children with sensorimotor disorders: a systematic review. AB - The aim of this review was to examine systematically the evidence for the application of interactive computer play in the rehabilitation of children with sensorimotor disorders. A literature search of 11 electronic databases was conducted to identify articles published between January 1995 and May 2008. The review was restricted to reports of intervention studies evaluating the impact of interactive computer play on motor rehabilitation in children. For each study the quality of the methods and the strength of the evidence were assessed by two independent reviewers using the guidelines of the American Academy for Cerebral Palsy and Developmental Medicine. A total of 74 articles were identified, of which 16 met the inclusion criteria. Three studies were randomized controlled trials (RCTs) and half were case series or case reports. Areas investigated were movement quality, spatial orientation and mobility, and motivational aspects. Thirteen studies presented positive findings. Two of the three RCTs investigating movement quality and one level III study examining spatial orientation showed no significant improvements. Interactive computer play is a potentially promising tool for the motor rehabilitation of children but the level of evidence is too limited to assess its value fully. Further and more convincing research is needed. PMID- 19191835 TI - Subjective well-being and satisfaction with life in adults with spastic cerebral palsy: a pilot study of a randomized sample. AB - The aim of this study was to explore subjective well-being and satisfaction with life in general and with participation, activities, and health, in adults with cerebral palsy (CP). A randomized sample of 50 individuals was drawn from a sample of 465 previous outpatients of a regional social paediatric centre in southern Germany. A psychological measure for well-being (Bern Questionnaire on Subjective Well-Being, adult form [BSW/A]) and a sociomedical measure (Life Satisfaction Checklist [LiSat-11]) were applied as outcome measures. Twenty-five individuals from 35 responders (nine males, 16 females; mean age 28y, SD 2y 7mo, range 25-33y) were evaluated completely. Nineteen participants had bilateral and six had unilateral spastic types of CP (Gross Motor Function Classification System Levels I n=5, II n=5, III n=5, IV n=8, and V n=2). Eighteen individuals had no or only minor intellectual disabilities. Subjective well-being and general satisfaction with life were not impaired in adults with CP. Joy of life (a subscale of the BSW/A) was even increased. Satisfaction with some areas of participation was reduced compared with a reference and clinical samples from Sweden. In conclusion, subjective well-being and general satisfaction with life are not decreased in adults with CP. The different levels of satisfaction with areas of participation in German and Swedish individuals with CP may be explained by differences of social inclusion. PMID- 19191836 TI - Delayed language development due to infantile thiamine deficiency. AB - The aim of this study was to investigate the language development of 20 children who had been exposed to thiamine (vitamin B(1)) deficiency in infancy due to feeding with soy-based formula that was accidentally deficient of thiamine. In this case-control study, 20 children (12 males, eight females; mean age 31.8mo [SD 4.1], range 24-39mo) who were fed thiamine-deficient formula in infancy were compared with 20 children (12 males, eight females; mean age 32.2mo [SD 3.9], range 25-39mo) fed with other milk sources and matched for age, sex, and maternal education. Receptive and expressive language development was assessed with the Preschool Language Scale, 3rd edition. Other assessments included mental development (Bayley Scales of Infant Development, 2nd edition), evaluation for autistic spectrum disorders, and neurological examination. Motor development was compared by age at independent walking. The study and control groups differed significantly in the expressive communication (p<0.001) and auditory comprehension language subscales (p<0.001), the Mental Developmental Index score (p<0.001), and age at independent walking (p=0.001). A significant correlation was found between the receptive language score and age at independent walking, i.e. poorer language associated with later walking (r=-0.601, p=0.005). The conclusion was that thiamine deficiency in infancy could affect language development in childhood. PMID- 19191837 TI - The constraints of terminology. PMID- 19191839 TI - Does botulinum toxin prevent or promote deformity in children with cerebral palsy? PMID- 19191840 TI - Thrombolysis in paediatric arterial ischaemic stroke. PMID- 19191841 TI - Measuring participation in childhood disability: how does the capability approach improve our understanding? PMID- 19191842 TI - Systematic reviews of psychosocial interventions for autism: an umbrella review. AB - AIM: A wide range of psychosocial interventions for the treatment of individuals with autism-spectrum disorders (ASDs) have been evaluated in systematic reviews. We conducted an umbrella review of systematic reviews of the effectiveness of psychosocial interventions for ASD. METHOD: Comprehensive searches were conducted in 25 bibliographic databases, relevant journals and reference lists up to May 2007. Studies included were systematic reviews on any psychosocial intervention for individuals with ASDs. Two reviewers independently assessed study relevance and quality. RESULTS: Thirty systematic reviews were included. The majority of reviews evaluated interventions based on behavioural theory (n=9) or communication-focused (n=7) therapies. Positive intervention outcomes were reported in the majority of the reviews. Methodological quality of the reviews was generally poor. INTERPRETATION: The reviews reported positive outcomes for many of the interventions, suggesting that some form of treatment is favourable over no treatment. However, there is little evidence for the relative effectiveness of these treatment options. Many of the systematic reviews had methodological weaknesses that make them vulnerable to bias. There is a need for further systematic reviews that adhere to strict scientific methods and for primary studies that make direct comparisons between different treatment options. PMID- 19191843 TI - Continuous postural management and the prevention of deformity in children with cerebral palsy: an appraisal. AB - Continuous postural management programmes are commonly used for children with cerebral palsy (CP) in Gross Motor Function Classification System levels IV and V, with the aim of preventing musculoskeletal deformity. There is a lack of evidence to support their use in this capacity and a possibility that children with CP who are most likely to develop deformity may be least able to comply with a continuous postural management programme. The implications for the child and family of such a programme in terms of increased demands and potential discomfort are discussed within the framework of the International Classification of Functioning, Disability and Health. A shift in focus in the use of postural management from an emphasis on body structure towards the environment and participation of the child with CP is suggested. PMID- 19191844 TI - A randomized controlled trial of the impact of therapeutic horse riding on the quality of life, health, and function of children with cerebral palsy. AB - This randomized controlled trial examined whether therapeutic horse riding has a clinically significant impact on the physical function, health and quality of life (QoL) of children with cerebral palsy (CP). Ninety-nine children aged 4 to 12 years with no prior horse riding experience and various levels of impairment (Gross Motor Function Classification System Levels I-III) were randomized to intervention (10wks therapeutic programme; 26 males, 24 females; mean age 7y 8mo [SD 2y 5mo] or control (usual activities, 27 males, 22 females; mean age 8y 2mo [SD 2y 6mo]). Pre- and post-measures were completed by 72 families (35 intervention and 37 control). Children's gross motor function (Gross Motor Function Measure [GMFM]), health status (Child Health Questionnaire [CHQ]), and QoL (CP QoL-Child, KIDSCREEN) were assessed by parents and QoL was assessed by children before and after the 10-week study period. On analysis of covariance, there was no statistically significant difference in GMFM, CP QoL-Child (parent report and child self-report), and CHQ scores (except family cohesion) between the intervention and control group after the 10-week study period, but there was weak evidence of a difference for KIDSCREEN (parent report). This study suggests that therapeutic horse riding does not have a clinically significant impact on children with CP. However, a smaller effect cannot be ruled out and the absence of evidence might be explained by a lack of sensitivity of the instruments since the QoL and health measures have not yet been demonstrated to be sensitive to change for children with CP. PMID- 19191845 TI - Long-term effects of botulinum toxin A in children with cerebral palsy. AB - The long-term effects of botulinum toxin A (BoNT-A) treatment in children with cerebral palsy (CP) are still elusive. We studied a prospective clinical cohort of 94 children with different subtypes (50% spastic diplegic CP, 22% hemiplegic CP, 25% tetraplegic CP, 3% dyskinetic CP), sex (55% male, 45% female), severity according to Gross Motor Function Classification System (29% Level I, 15% Level II, 16% Level III, 17% Level IV, 23% Level V), and age (median 5y 4mo, range 11mo 17y 8mo). The longest follow-up time was 3 years 7 months (median 1y 6mo) and included a maximum of eight injections per muscle (median two injections to a specific muscle). Outcome measurements were muscle tone (Modified Ashworth Scale) and joint range of motion (ROM). Assessments were made at a minimum before and 3 months after each injection. Ninety-five per cent confidence intervals for differences from baseline were used to identify significant changes. BoNT-A injections induced reduction of long-term spasticity in all muscle-groups examined: the gastrocnemius, hamstring, and adductor muscles. The reduction in tone was most distinct in the gastrocnemius muscle, and each repeated injection produced an immediate reduction in muscle tone. However, improvement in ROM was brief and measured only after the first injections, whereupon the ROM declined. Thus, the results suggest that BoNT-A can be effective in reducing muscle tone over a longer period, but not in preventing development of contractures in spastic muscles. The dissociation between the effects on muscle tone and ROM indicates that development of contractures is not coupled to increased muscle tone only, but might be caused by other mechanisms. PMID- 19191846 TI - Bacterial endocarditis in a child presenting with acute arterial ischemic stroke: should thrombolytic therapy be absolutely contraindicated? AB - Thrombolysis is considered to be contraindicated in acute ischemic stroke secondary to infective endocarditis (IE). We report a 12-year-old female who presented with acute dense right hemiparesis and aphasia. Cranial magnetic resonance imaging and angiography showed multiple diffusion-restricted lesions in the left hemisphere and absence of flow in the left internal carotid artery. She was treated with intra-arterial tissue plasminogen activator within 6 hours of her presentation. Subsequently she was diagnosed with pneumococcal endocarditis and underwent debridement of vegetations and patch repair of the mitral valve. The patient did not have hemorrhagic complications following thrombolytic therapy or surgery. Pathological analysis of the mitral valve vegetations revealed mostly fibrin thrombus. Follow-up imaging showed complete recanalization of the left internal carotid artery, and the patient had a remarkable neurological recovery. This is the first case report of successful intra-arterial thrombolytic therapy in childhood IE-related stroke. We believe that thrombolytic therapy contributed to a favorable outcome in our patient and may be safe in selected patients with childhood IE-related acute ischemic stroke. PMID- 19191847 TI - Local intra-arterial thrombolysis in a 4-year-old male with vertebrobasilar artery thrombosis. AB - We report the case of a 4-year-old male with vertebrobasilar artery thrombosis for which he was treated with local intra-arterial urokinase 60 hours after onset of symptoms. Initially the patient had dysarthria and dysphagia. Brain magnetic resonance imaging (MRI) in a community hospital showed abnormalities in the pons, and vertebrobasilar artery thrombosis was suspected. The patient was transferred to the university hospital because of neurological deterioration. Just before thrombolysis, his physical examination revealed a downward ocular deviation, stretching of the arms, and spontaneous movements of the legs. Brain MRI showed recent ischaemia in the pons and caudal part of the mesencephalon, and magnetic resonance angiography showed occlusion of the basilar artery. Intra-arterial thrombolysis with urokinase (total dose 36 000U/kg) was performed 60 hours after onset of symptoms. After thrombolysis he was treated with heparin for 10 days, followed by aspirin. The patient recovered well. After 1 year he had only minor ataxia and performed all activities at school. Local vasculitis due to an infection with Borrelia burgdorferi was thought to be the cause of the local thrombosis. A review of the literature revealed 11 other case reports of successful local intra-arterial thrombolysis in children and adolescents with ischaemic stroke. This suggests that intra-arterial thrombolysis is feasible and safe in selected paediatric patients with ischaemic stroke. PMID- 19191848 TI - Rituximab as a highly effective treatment in a female adolescent with severe multiple sclerosis. PMID- 19191849 TI - Prognostic factors of radiotherapy in patients with node-positive thoracic esophageal squamous cell carcinoma after radical surgery. AB - The aim of this study was to retrospectively analyze and assess the outcomes and prognostic factors of radiotherapy in patients with node-positive thoracic esophageal squamous cell carcinoma after radical surgery. One hundred twenty-six patients with node-positive thoracic esophageal squamous cell carcinoma who had undergone adjuvant therapy (postoperative radiotherapy alone or postoperative sequential chemoradiotherapy without receiving postoperative concurrent chemoradiotherapy) after radical surgery, were retrospectively reviewed from January 1996 to December 2003. Univariate and multivariate analyses were performed using log-rank and Cox proportional hazard models, and survival curves were estimated using the Kaplan-Meier method. The 1-, 3- and 5-year overall survival rates of all 126 patients were 71.4, 39.1, and 22.0%, and disease-free survival rates were 64.3, 36.4, and 21.5%, respectively. Lymph node ratio (the ratio of the number of metastatic lymph nodes to the number of lymph nodes removed, LNR) > or = 0.2 (P= 0.006), pT3 + pT4 (P= 0.06) and sequential chemoradiotherapy (P= 0.08) were associated with a poorer survival by univariate analysis. In multivariate analysis, LNR (P= 0.01, hazard ratio = 0.57, 95% confidence interval, 0.37-0.87) and tumor depth of invasion (P= 0.03, hazard ratio = 0.62, 95% confidence interval, 0.41-0.96) were the independent predictors of survival. Sequential chemoradiotherapy receded survival tendency without significant difference (P= 0.09, hazard ratio = 0.64, 95% confidence interval, 0.37-1.08). Therefore, LNR and tumor depth of invasion were the independent prognostic factors of radiotherapy in patients with node-positive thoracic esophageal squamous cell carcinoma after radical surgery. The addition of chemotherapy does not seem to confer a survival benefit. PMID- 19191850 TI - Efficacy of chemoradiotherapy with low-dose cisplatin and continuous infusion of 5-fluorouracil for unresectable squamous cell carcinoma of the esophagus. AB - We retrospectively investigated the efficacy of a chemoradiotherapy regimen using daily low-dose cisplatin and continuous 5-fluorouracil infusion in 71 registered patients with unresectable esophageal cancer. The overall response rate (complete response plus partial response) was 59%. The major toxicities observed were leukopenia and anorexia. The 1- and 3-year overall survival rates were 54.6% and 18.4%, respectively. A low preoperative C-reactive protein level was found to be associated with a good response. The pretreatment performance status and response results were both shown to be prognostic factors for overall survival. These findings confirmed that the chemoradiotherapy regimen had curative potential for unresectable esophageal cancer. PMID- 19191851 TI - Endoscopy of the esophagus in gastroesophageal reflux disease: are we losing sight of symptoms? Another perspective. AB - Gastroesophageal reflux disease (GERD) is an extremely common chronic disorder associated with impaired quality of life and huge economic burden. Recently, an International Consensus Group developed a global definition of GERD (The Montreal Definition): a condition that develops when the reflux of stomach contents causes troublesome symptoms and/or complications. The traditional endoscopy-based classification of GERD patients into one of three groups - non-erosive reflux disease, erosive esophagitis, and Barrett's esophagus - is fraught with several limitations. Due to the lack of a gold standard, GERD is a symptom-based diagnosis, and hence symptom evaluation will remain the primary means by which treatment decisions are made for patients with suspected GERD. We propose that patients reporting the predominant GERD-like symptoms (GERS) in the primary care setting be classified based upon their response to an empiric trial of acid suppressive therapy: complete response to acid suppressive therapy, partial response to acid suppressive therapy, and no response to acid suppressive therapy. Given the limitations of objective medical testing, implementation of our proposed new symptom-based classification of patients with GERS would guide primary care physicians on when to refer patients to a gastroenterologist, which in turn could help in better resource utilization. Validation of this proposed classification by well-designed prospective multicenter studies is awaited. PMID- 19191852 TI - Influence of Helicobacter pylori eradication on reflux esophagitis in Japanese patients. AB - The relationship between Helicobacter pylori eradication and reflux esophagitis is controversial. We analyzed the development of reflux esophagitis and the change in the grade of pre-existing reflux esophagitis after eradication. Enrolled were 559 Japanese patients who received eradication therapy for H. pylori. The grade of reflux esophagitis by endoscopy before and after therapy was evaluated retrospectively. No esophagitis was present before eradication in 526 patients. H. pylori was and was not eradicated in 429 and 97, respectively. Reflux esophagitis developed in 40 of the eradication group and in three of the treatment failure group, with prevalence higher with successful eradication (P = 0.04). Successful eradication and hiatus hernia were significant risk factors for reflux esophagitis development. Twenty-seven of 33 patients with pre-existing reflux esophagitis had successful eradication and six treatment failure. The reflux esophagitis grade worsened in two (Los Angeles classification from A to B) and improved in 14 patients after eradication. With treatment failure, reflux esophagitis worsened in none and improved in three patients. There showed no significant change in the grade of pre-existing reflux esophagitis after H. pylori eradication but the sample size was too small to evaluate the difference. In conclusion, the eradication of H. pylori increases the prevalence of reflux esophagitis, and hiatus hernia was a significant risk factor for the development of reflux esophagitis. PMID- 19191853 TI - A study comparing tolerability, satisfaction and acceptance of three different techniques for esophageal endoscopy: sedated conventional, unsedated peroral ultra thin, and esophageal capsule. AB - Three methods of esophagoscopy are available until now: sedated conventional endoscopy, unsedated ultrathin endoscopy, and esophageal capsule endoscopy. The three methods carry comparable diagnostic accuracy and different complication rates. Although all of them have been found well accepted from patients, no comparative study comprising the three techniques has been published. The aim of this study was to compare the three methods of esophagoscopy regarding tolerability, satisfaction, and acceptance. Twenty patients with large esophageal varices and 10 with gastroesophageal reflux disease were prospectively included. All patients underwent consecutively sedated conventional endoscopy, unsedated ultrathin endoscopy, and esophageal capsule endoscopy. After each procedure, patients completed a seven-item questionnaire. The total positive attitude of patients toward all methods was high. However, statistical analysis revealed the following differences in favor of esophageal capsule endoscopy: (i) total positive attitude has been found higher (chi(2)= 18.2, df = 2, P= 0.00), (ii) less patients felt pain (chi(2)= 6.9, df = 2, P= 0.03) and discomfort (chi(2)= 22.1, df = 2, P= 0.00), (iii) less patients experienced difficulty (chi(2)= 13.7, df = 2, P= 0.01), and (iv) more patients were willing to undergo esophageal capsule endoscopy in the future (chi(2)= 12.1, df = 2, P= 0.002). Esophageal capsule endoscopy was characterized by a more positive general attitude and caused less pain and discomfort. Sedated conventional endoscopy has been found more difficult. More patients would repeat esophageal capsule endoscopy in the future. Patients' total position for all three available techniques for esophageal endoscopy was excellent and renders the observed advantage of esophageal capsule endoscopy over both sedated conventional and unsedated ultrathin endoscopy a statistical finding without a real clinical benefit. PMID- 19191854 TI - Self-expanding plastic stents for inoperable malignant strictures of the cervical esophagus. AB - Dysphagia and respiratory complications are the major problems in patients suffering from malignant strictures of the cervical esophagus. In inoperable cases, interventional palliation is the cornerstone of treatment. The aim of this study was to evaluate the use of self-expanding plastic stents (SEPS) in this group of patients. In a retrospective study, 23 patients suffering from various malignant obstructive diseases of the cervical esophagus, including squamous cell carcinoma (n = 10), laryngeal cancer (n = 7), lung cancer with esophageal invasion (n = 5), and metastatic breast cancer (n = 1), underwent SEPS placement, under endoscopic and fluoroscopic guidance. Tracheoesophageal fistula was documented in five patients. Technical success rate, improvement of dysphagia grade, and stent-related complications were evaluated after stent placement. Stent insertion was successfully achieved in all cases, namely in 20 patients at the first stent placement attempt and in three patients after a second attempt. Dysphagia grade was notably improved after 24 h. In two cases, major complications occurred. These were successfully treated without the need of stent extraction. Barium swallowing studies demonstrated complete sealing of all fistulas. Foreign-body sensation that gradually disappeared within the first week after stent placement was observed in eight patients. Recurrence of dysphagia occurred in three patients, due to hyperplastic tissue proliferation (n = 2) and tumor overgrowth (n = 1). Late migration of the stent was detected in one case after 67 days. SEPS placement is an effective and safe palliative treatment for malignant strictures of the cervical esophagus. Main advantages include easy retrievability and reduced rates of reinterventions. PMID- 19191856 TI - International survey on esophageal cancer: part I surgical techniques. AB - In patients with esophageal cancer, radical surgical resection of the esophagus and surrounding lymph nodes is the only curative treatment option. Nevertheless, no standard surgical procedure exists. The aims of the present study were to gain insight into the frequencies of the various surgical techniques in esophageal cancer surgery as applied by surgeons throughout the world and to identify intercontinental differences regarding surgical techniques. Surgeons with particular interest in esophageal surgery, including members of the International Society for Diseases of the Esophagus, the European Society of Esophagology Group d'Etude Europeen des Maladies de l'Oesophage and the OESO, were invited to participate in an online questionnaire. Questions were asked regarding approach to esophagectomy, extent of lymphadenectomy (LND), type of reconstruction, and anastomotic techniques. Subanalyses were performed for the surgeons' case volume per year, years of experience in esophageal cancer surgery, and continent. Of 567 invited surgeons, 269 participated, resulting in an overall response rate of 47%. The responders currently performing esophagectomies (n= 250; 44%), represented 41 countries across the six continents. Fifty-two percent of responders favor open transthoracic esophagectomy (TTE) over transhiatal esophagectomy (THE) or minimally invasive esophagectomy (MIE). THE is preferred by 26%, whereas MIE is favored by 14%. Eight percent have no preference for one approach to esophagectomy over the other. The extent of LND is most frequently the 2-field, routinely performed by 73% of surgeons. The continuity of the digestive tract is most frequently restored with a gastric conduit (85%). In open TTE, the anastomosis is routinely created in the neck by 56% of responders and in the chest by 40%. Cervical anastomoses are routinely fashioned by means of a handsewn technique by 65% of responders, while 35% favor the stapled technique. The cervical incision is predominantly performed vertically on the left side of the neck (routinely by 66%). A horizontal neck incision is routinely carried out by 19% of responders and a vertical right-sided incision by 11%. Significant differences in surgical techniques could be detected between low- and high-volume surgeons, between surgeons with or=21 years of experience, and between surgeons from different continents. In conclusion, currently the most commonly applied surgical procedure is the open right-sided transthoracic approach with a two-field lymphadenectomy, using a gastric tube anastomosed at the left side of the neck by means of a handsewn, end-to-side technique. The results of this survey provide baseline data for future research and for the development of international guidelines. PMID- 19191855 TI - International survey on esophageal cancer: part II staging and neoadjuvant therapy. AB - The outcome of esophagectomy could be improved by optimal diagnostic strategies leading to adequate preoperative patient selection. Neoadjuvant therapy could improve outcome by increasing the number of radical resections and by controlling metastatic disease. The purposes of this study were to gain insight into the current worldwide practice of staging modalities and neoadjuvant therapy in esophageal cancer, and to detect intercontinental differences. Surgeons with particular interest in esophageal surgery, including members of the International Society for Diseases of the Esophagus, the European Society of Esophagology - Group d'Etude Europeen des Maladies de l'Oesophage, and the OESO, were invited to participate in an online questionnaire. Questions were asked regarding staging modalities, neoadjuvant therapy, and response evaluation applied in esophageal cancer patients. Of 567 invited surgeons, 269 participated resulting in a response rate of 47%. The responders currently performing esophagectomies (n= 250; 44%) represented 41 countries across the six continents. Esophagogastroscopy with biopsy and computed tomography (CT) scanning were routinely performed by 98% of responders for diagnosing and staging esophageal cancer, while endoscopic ultrasound (EUS) and barium esophagography were routinely applied by 58% and 51%, respectively. Neoadjuvant therapy is routinely administered by 33% and occasionally by 63% of responders. Of the responders that administer identical neoadjuvant regimens to esophageal adenocarcinoma (AC) and squamous cell carcinoma, 54% favor chemoradiotherapy. For AC, chemotherapy is preferred by 31% of the responders that administer neoadjuvant therapy, whereas for squamous cell carcinoma, the majority of responders (38%) prefer chemoradiotherapy. Response to neoadjuvant therapy is predominantly assessed by CT scanning of the chest and abdomen (86%). Barium esophagography, EUS, and combined CT/PET scan are requested for response monitoring in equal frequency (25%). Substantial differences in applied staging modalities and neoadjuvant regimens were detected between surgeons from different continents. In conclusion, currently the most commonly applied diagnostic modalities for staging and restaging esophageal cancer are CT scanning of the chest and abdomen, gastroscopy, barium esophagography and EUS. Neoadjuvant therapy is routinely applied by one third of the responders. Intercontinental differences have been detected in the diagnostic modalities applied in esophageal cancer staging and in the administration of neoadjuvant therapy. The results of this survey provide baseline data for future research and for the development of international guidelines. PMID- 19191857 TI - Overexpression of MMP-2 and MMP-9 in esophageal squamous cell carcinoma. AB - Matrix metalloproteinases (MMPs) are known to play important roles in extracellular matrix remodeling during the process of tumor invasion and metastasis. However, little is known about their role in esophageal squamous cell carcinoma (ESCC). Expression of MMP-2 and MMP-9 in ESCC was detected in our research. Tissue microarray chip was prepared, consisting of 58 cases of ESCC and corresponding esophageal epithelium tissues. MMP-2 and MMP-9 were examined by immunohistochemistry. Overexpression of MMP-2 and MMP-9 was found in ESCC (42.1 and 60.3%, respectively), compared with paired distal normal esophageal tissues (22.9 and 8.9%, respectively). Expression of MMP-2 in ESCC was significantly associated with the tumor invasion depth, tumor-node-metastasis stages, and lymph node metastasis. MMP-2 and MMP-9 may play important roles in carcinogenesis, and MMP-2 may act as a biological marker of invasion and lymph node metastasis in ESCC. PMID- 19191858 TI - Management of delayed intrathoracic esophageal perforation with modified intraluminal esophageal stent. AB - In this article, we reviewed our experience of treatment of the delayed intrathoracic nonmalignant esophageal perforation employing modified intraluminal esophageal stent. Between February 1990 and August 2006, eight patients were included in this study. Five patients experienced sepsis. The interval time between perforation and stent placement ranged from 36 h to 27 days (average, 8.6 days). Esophageal stenting and throracotomy for foreign body removal were performed in four patients. The remaining four patients underwent stent placement and thoracostomy. Nutrition was initiated through gastrostomy after 7 to 10 days after the stenting. The stent was removed after the patients resumed oral intake of food and the esophagogram showed that perforation was closed. There was no death in this group. Signs of sepsis remitted 1 week after stent placement. Complications included stress ulcer, stimulative cough, and pneumonia each. Stent removal ranged 32 to 120 days (average 66.7) after its placement. The stent was kept in place for 4 months to prevent formation of esophageal stricture in one patient with caustic esophageal burns. The follow-up was completed in all the patients. The mean follow-up period was 59 months (range 12-180). One patient with caustic esophageal burn underwent cicatricial esophagectomy and gastric transposition 3 years later due to the esophageal stricture. Barium swallow demonstrated that there was a diverticulum-like outpouching in one patient and slight esophageal stricture at T2 and T3 level in another. One patient developed reflux esophagitis 5 years after stent removal. All the patients finally had a normal intake of food. Modified esophageal stenting is an effective method to manage the delayed intrathoracic esophageal perforation. Prevention of stent migration and its convenient adjustment might be the major advantages of this method. PMID- 19191859 TI - Surgical repair of refractory strictures of esophagogastric anastomoses caused by leakage following esophagectomy. AB - Refractory strictures of esophagogastric anastomosis caused by leakage following an esophagectomy are a severe complication, for which either repeated balloon dilations or bougies are not necessarily effective. In such a case, surgical repair is quite difficult because the esophageal substitute such as the stomach or colon is usually located in the mediastinum and severely adhesive to the neighboring organs. Furthermore, in case the resected stricture is too long for direct re-anastomosis to be performed, a free jejunal graft or a new esophageal substitute should be prepared. This paper proposes a procedure for the re reconstruction of refractory stricture in the case of a retrosternal reconstruction with a gastric conduit, which frequently employs pull-up route. The anterior plate of the manubrium was divided medially from the notch to the symphysis with the sternal saw. The manubrium is then removed, bite by bite, like breaking up rocks, with a bone rongeur forceps, starting with the anterior plate, then the posterior plate, from upper median part to the lower and lateral part of the sternum until it reaches the symphysis and the sternoclavicular and the sternocostal joints. It is safer to destroy the manubrium little by little from the anterior side so that the posterior periosteum, which is likely to adhere tightly to the gastric conduit, can be preserved. After the manubrium is almost completely resected and the posterior periosteum of the manubrium is preserved, a median longitudinal incision is carefully made on the periosteum so as not to damage the gastric conduit that may be adhesive to the periosteum. The periosteum was gradually opened bilaterally separating the periostium and the gastric conduit. Although gastroenterological surgeons may hesitate to remove the manubrium, removing the manubrium and preserving the posterior periosteum make it possible to avoid injuring the gastric conduit and to provide a wide view around the stenosis for safely resecting the anastomotic stricture. Furthermore, this procedure allows direct re-anastomosis between the cervical esophagus and the gastric conduit without a complicated reconstruction such as a free jejunal graft. This procedure is strongly recommended as an alternative option so that a second reconstruction can be performed both safely and steadily. PMID- 19191860 TI - Assessment of novel endoscopic techniques for visualizing superficial esophageal squamous cell carcinoma: autofluorescence and narrow-band imaging. AB - Lugol chromoendoscopy (LCE) is a useful technique for visualizing superficial esophageal squamous cell carcinoma (SESCC), but the stimulating effect of the Lugol solution can sometimes cause clinical problems. Newly developed techniques such as narrow-band imaging (NBI) and autofluorescence imaging (AFI) enable SESCC to be easily visualized without LCE. This study aimed to assess the visualizing power of white-light imaging (WLI), NBI, and AFI, compared with LCE. Sixteen patients with 16 SESCCs underwent LCE and endoscopy with NBI and AFI before endoscopic or surgical treatment. Twenty sets of endoscopic SESCC images were prepared, each of which contained still images from WLI, NBI, AFI, and LCE. The image sets were shown to 25 endoscopists, who then each completed a questionnaire about the ease-of-detection of the SESCCs, scoring WLI, NBI, and AFI images with reference to a perfect score for LCE; mean scores were compared. Overall, significantly higher scores were given for NBI than for WLI and AFI, with no significant difference between WLI and AFI. Stratification by endoscopist characteristics indicated that younger or less experienced endoscopists gave significantly higher scores for AFI than WLI. Stratification by lesion characteristics revealed that AFI had significantly higher scores than WLI for flat/elevated lesions or those with diameter >or=20 mm; scores were significantly lower for depressed lesions or those with diameter <20 mm. For SESCC, the visualizing power of NBI seems more similar to that of LCE than AFI or WLI: NBI might be more useful than AFI or WLI in detecting SESCC. AFI seems to have both superior and inferior visualizing power to WLI depending on characteristics of endoscopists or SESCC lesions. PMID- 19191861 TI - The characteristics of Barrett's esophagus: an analysis of 4120 cases in China. AB - Our objective was to investigate the endoscopic and clinico-pathological characteristics in patients with Barrett's esophagus (BE) in China. Using the terms 'Barrett's esophagus' and 'Barrett's esophagus, China' as key words, literatures published in Chinese and English journals were searched in Chinese data banks, as well as PubMed and ISI Web of Science from 1989 to 2007. An analysis was carried out with the standard inclusion and exclusion criteria. A total of 4120 cases were included in this study. BE was found in 2.44% of patients undergoing endoscopy for various symptoms of upper gastrointestinal tract diseases; the male : female ratio was 2.09 : 1, the average age of detection of BE was 53.15 years old, and 51% of patients with BE had typical symptoms for gastroesophageal reflux disease (GERD). The island-type BE was predominant (56.80%), and the occurrence of BE with special intestinal metaplasia (SIM) was 36.58%, but SIM was more common in tongue-type BE than island-type and circumferential-type BE (both P < 0.001), as well as in long segment BE (LSBE) than in short segment BE (SSBE) (P < 0.001). A total of 46.39% of patients had Helicobacter pylori infection. The mean length of follow up was 2 years in 492 patients. The incidence of adenocarcinoma was 0.61% patient-years of total follow up. In China, the endoscopic prevalence of BE is lower, but the average age of diagnosis is younger; a high proportion of H. pylori infection is found in patients with BE, and about half of the patients have no typical symptoms of GERD; the tongue-type BE and the LSBE are apt to SIM. PMID- 19191862 TI - Surgical results of patients after esophageal resection or extended gastrectomy for cancer of the esophagogastric junction. AB - Precise classification of cancers of the esophagogastric junction according to Siewert may be difficult for the presence of Barrett's esophagus or hiatal hernia, which subsequently leads to a difficult choice of the surgical procedure of esophagectomy or gastrectomy. Ninety-six patients with such cancers were operated on in our department in 7 years. Twenty-nine patients (30.2%), classified as type I (group 1), underwent a transthoracic esophagectomy with gastric pull up. Sixty-seven patients (69.8%) classified as type II or III (group 2) underwent an extended gastrectomy. We compared the patients of both groups retrospectively for disease-free survival and postoperative complications. The general performance status of most patients was comparable in both groups and was assigned to the American Society of Anesthesiologists class II or III. Statistically significant differences between the groups were seen for the postoperative reintubation rate [group 1: 31.0% vs. group 2: 9.0% (P = 0.009)], median time for surgery [group 1: 6 (3.5-8.5) hours vs. group 2: 4.7 (2.2-11.5) hours (P = 0.001)], time in the intensive care unit [group 1: 6 (3-85) days vs. group 2: 3 (1-54) days (P = 0.001)], median hospitalization time [group 1: 23 (14 105) days vs. group 2: 18 (10-63) days (P = 0.018)]. No statistical difference was observed for the recurrence-free survival of 40% after 3 years (P = 0.311), the mortality rate, the morbidity rate (P = 0.108), surgical and respiratory complications, and the incidence of anastomotic leakage (P = 0.645). We conclude that in selected cases it may be possible to perform an extended gastrectomy for small type I cancers. PMID- 19191863 TI - Reduction in labile plasma iron during treatment with deferasirox, a once-daily oral iron chelator, in heavily iron-overloaded patients with beta-thalassaemia. AB - This subgroup analysis evaluated the effect of once-daily oral deferasirox on labile plasma iron (LPI) levels in patients from the prospective, 1-yr, multicentre ESCALATOR study. Mean baseline liver iron concentration and median serum ferritin levels were 28.6 +/- 10.3 mg Fe/g dry weight and 6334 ng/mL respectively, indicating high iron burden despite prior chelation therapy. Baseline LPI levels (0.98 +/- 0.82 micromol/L) decreased significantly to 0.12 +/ 0.16 micromol/L, 2 h after first deferasirox dose (P = 0.0006). Reductions from pre- to post-deferasirox administration were also observed at all other time points. Compared to baseline, there was a significant reduction in preadministration LPI that reached the normal range at week 4 and throughout the remainder of the study (P < or = 0.02). Pharmacokinetic analysis demonstrated an inverse relationship between preadministration LPI levels and trough deferasirox plasma concentrations. Once-daily dosing with deferasirox > or =20 mg/kg/d provided sustained reduction in LPI levels in these heavily iron-overloaded patients, suggesting 24-h protection from LPI. Deferasirox may therefore reduce unregulated tissue iron loading and prevent further end-organ damage. PMID- 19191864 TI - Identification of the first in cis mutations in MYH9 disorder. AB - Here, we report the first in cis mutations in exon 1 of the MYH9 gene in a patient with MYH9 disorder. The patient was a 5-yr-old girl with macrothrombocytopenia and conspicuous cytoplasmic inclusion bodies in neutrophils. Immunofluorescence analysis of neutrophil non-muscle myosin heavy chain-II A (NMMHC-IIA) indicated several cytoplasmic spots of NMMHC-IIA aggregates that were circular to oval in shape (type II pattern). Mutational analysis showed two mutations, c.99G > T and c.103C > G, which would result in p.W33C and p.P35A, respectively, in exon 1 of the MYH9 gene. In addition, concurrent mutations were present on the same chromosome. Inclusion bodies are usually faint or mostly invisible in MYH9 disorders with a mutation in exon 1. In this case, double mutations might have caused the large myosin protein aggregation and accumulation. Although not observed in this patient, the development of Alport manifestations should be monitored by careful follow-up. PMID- 19191865 TI - Cerebral aspergillosis. PMID- 19191866 TI - Anti-thymocyte globulin therapy induced a spurious increase of fibrinogen degradation products in hypoplastic myelodysplastic syndrome. PMID- 19191867 TI - Specific assessment of BCR-ABL transcript overexpression and imatinib resistance in chronic myeloid leukemia patients. AB - Imatinib mesylate has proven to be the most effective treatment in chronic myeloid leukemia. Nevertheless, imatinib resistance has raised concern and prompted interest in additional strategies to achieve disease eradication. Resistance to imatinib is mainly associated with three mechanisms: acquired mutations in the kinase domain of BCR-ABL protein, genetic amplification, and transcript overexpression of BCR-ABL rearrangement. Therefore an accurate assessment of resistance mechanism is particularly important to improve strategies to overcome resistance. In order to determine overexpression of BCR ABL, we propose a method that correlates quantitative real time PCR and fluorescence in situ hybridization data from the same peripheral blood sample. The ratio between both methodologies permits to calculate the expression index (EI) for each patient. EI estimates the rate of BCR-ABL transcription per rearrangement. The median EI value, including all cases (n = 123), was 0.288; those cases (n = 13) included in percentile 90 showed an increment of EI above 1 Log (>2.88) with respect to the median value and were considered as cases with overexpression. We also evaluated the EIs using receiver operating characteristics curve; choosing an EI cutoff of 1.836 we obtained a sensitivity of 95% and a specificity of 61%. Using this EI cutoff value, more patients (n = 17) were included in the overexpression group. Patients within this group were resistant to imatinib and also showed a worse overall survival if compared with the remaining. PMID- 19191869 TI - Bovicin HC5 inhibits wasteful amino acid degradation by mixed ruminal bacteria in vitro. AB - Streptococcus bovis HC5 produces a broad spectrum lantibiotic (bovicin HC5) that inhibits pure cultures of hyper ammonia-producing bacteria (HAB). Experiments were preformed to see if: (1) S. bovis HC5 cells could inhibit the deamination of amino acids by mixed ruminal bacteria taken directly from a cow, (2) semi purified bovicin was as effective as S. bovis HC5 cells, and 3) semi-purified and the feed additive monensin were affecting the same types of ammonia-producing ruminal bacteria. Because purified and semi-purified bovicin HC5 was as effective as S. bovis HC5 cells, it appeared that bovicin HC5 was penetrating the cell membranes of HAB before it could be degraded by peptidases and proteinases. Mixed ruminal bacteria that were successively transferred and enriched nine times with trypticase did not become significantly more resistant to either bovicin HC5 (50 AU mL(-1)) or monensin (5 microM), and amplified rDNA restriction analysis indicated that bovicin HC5 and monensin appeared to be selecting against the same types of bacteria. PMID- 19191868 TI - High expression of BCL3 in human myeloma cells is associated with increased proliferation and inferior prognosis. AB - BACKGROUND: BCL3 is a putative oncogene encoding for a protein belonging to the inhibitory kappaB-family. We experienced that this putative oncogene was a common target gene for growth-promoting cytokines in myeloma cell lines. METHODS: Gene expression of BCL3 was studied in 351 newly diagnosed myeloma patients, 12 patients with smouldering myeloma, 44 patients with monoclonal gammopathy of undetermined significance and 22 healthy individuals. Smaller material of samples was included for mRNA detection by RT-PCR, protein detection by Western blot and immunohistochemistry, and for cytogenetic studies. A total of eight different myeloma cell lines were studied. RESULTS: Bcl-3 was induced in myeloma cell lines by interleukin (IL)-6, IL-21, IL-15, tumor necrosis factor-alpha and IGF-1, and its upregulation was associated with increased proliferation of the cells. In a population of 351 patients, expression levels of BCL3 above 75th percentile were associated with shorter 5-yr survival. When this patient population was divided into subgroups based on molecular classification, BCL3 was significantly increased in a poor risk subgroup characterized by overexpression of cell cycle and proliferation related genes. Intracellular localization of Bcl-3 was dependent on type of stimulus given to the cell. CONCLUSION: BCL3 is a common target gene for several growth-promoting cytokines in myeloma cells and high expression of BCL3 at the time of diagnosis is associated with poor prognosis of patients with multiple myeloma (MM). These data may indicate a potential oncogenic role for Bcl-3 in MM. PMID- 19191870 TI - Influence of carbon monoxide on metabolite formation in Methanosarcina acetivorans. AB - Methanogenic archaea conserve energy for growth by reducing some one- and two carbon compounds to methane and concomitantly generating an ion motive force. Growth of Methanosarcina acetivorans on carbon monoxide (CO) is peculiar as it involves formation of, besides methane, formate, acetate and methylated thiols. It has been argued that methane formation is partially inhibited under carboxidotrophic conditions and that the other products result from either detoxification of CO or from bypassing methanogenesis with other pathways for energy conservation. To gain a deeper understanding of the CO-dependent physiology of M. acetivorans we analyzed metabolite formation in resting cells. The initial rates of methane, acetate, formate, and dimethylsulfide formation increased differentially with increasing CO concentrations but were maximal already at the same moderate CO partial pressure. Strikingly, further increase of the amount of CO was not inhibitory. The maximal rate of methane formation from CO was approximately fivefold lower than that from methanol, consistent with the previously observed significant downregulation of the energy converting sodium dependent methyltransferase. The rate of dimethylsulfide formation from CO was only 1-2% of that of methane formation under any conditions tested. Implications of the data presented for previously proposed pathways of CO utilization are discussed. PMID- 19191871 TI - Antimicrobial properties of resident coral mucus bacteria of Oculina patagonica. AB - The inhibitory properties of the microbial community of the coral mucus from the Mediterranean coral Oculina patagonica were examined. Out of 156 different colony morphotypes that were isolated from the coral mucus, nine inhibited the growth of Vibrio shiloi, a species previously shown to be a pathogen of this coral. An isolate identified as Pseudoalteromonas sp. was the strongest inhibitor of V. shiloi. Several isolates, especially one identified as Roseobacter sp., also showed a broad spectrum of action against the coral pathogens Vibrio coralliilyticus and Thallassomonas loyana, plus nine other selected Gram-positive and Gram-negative bacteria. Inoculation of a previously established biofilm of the Roseobacter strain with V. shiloi led to a 5-log reduction in the viable count of the pathogen within 3 h, while inoculation of a Pseudoalteromonas biofilm led to complete loss of viability of V. shiloi after 3 h. These results support the concept of a probiotic effect on microbial communities associated with the coral holobiont. PMID- 19191872 TI - Investigating the effects of positive charge and hydrophobicity on the cell selectivity, mechanism of action and anti-inflammatory activity of a Trp-rich antimicrobial peptide indolicidin. AB - To investigate the effects of positive charge and hydrophobicity on the cell selectivity, mechanism of action and anti-inflammatory activity of a Trp-rich antimicrobial peptide indolicidin (IN), a series of IN analogs with Trp-->Lys substitution were synthesized. All IN analogs displayed an approximately 7- to 18 fold higher cell selectivity, compared with IN. IN, IN-1 and IN-2 depolarized (50 90%) the cytoplasmic membrane potential of Staphylococcus aureus close to minimal inhibitory concentration (5-10 microg mL(-1)). However, other IN analogs (IN-3 and IN-4) displayed very low ability in membrane depolarization even at 40 microg mL(-1). Confocal laser-scanning microscopy revealed that IN-3 and IN-4 penetrated the Escherichia coli cell membrane, whereas IN, IN-1 and IN-2 did not enter the cell membrane. In the gel retardation assay, IN-3 and IN-4 bound more strongly to DNA compared with IN, IN-1 and IN-2. These findings suggest that the mechanism of antimicrobial action of IN-3 and IN-4 may be involved in the inhibition of intracellular functions via interference with DNA/RNA synthesis. Unlike IN, all IN analogs did not inhibit nitric oxide production or inducible nitric oxide synthase mRNA expression in lipopolysaccharide-stimulated mouse macrophage RAW264.7 cells, indicating that the hydrophobicity of IN is more important for anti-inflammatory activity in lipopolysaccharide-treated macrophage cells than the positive charge. PMID- 19191873 TI - Expression of the phosphonoalanine-degradative gene cluster from Variovorax sp. Pal2 is induced by growth on phosphonoalanine and phosphonopyruvate. AB - The phosphonopyruvate hydrolase (PalA) found in Variovorax sp., Pal2, is a novel carbon-phosphorus bond cleavage enzyme, which is expressed even in the presence of high levels of phosphate, thus permitting phosphonopyruvate to be used as the sole carbon and energy source. Analysis of the regions adjacent to the palA gene revealed the presence of the five structural genes that constitute the 2-amino-3 phosphonopropionic acid (phosphonoalanine)-degradative operon. Reverse transcriptase-PCR (RT-PCR) experiments demonstrated that all five genes in the operon are transcribed as a single mRNA and that their transcription is induced by phosphonoalanine or phosphonopyruvate. Transcriptional fusions of the regulatory region of the phosphonoalanine degradative operon with the gfp gene were constructed. Expression analysis indicated that the presence of a LysR-type regulator (encoded by the palR gene) is essential for the transcription of the structural genes of the operon. Similar gene clusters were found in the sequenced genomes of six bacterial species from the Alpha-, Beta- and Gammaproteobacteria, and analysis of metagenomic libraries revealed that sequences related to palA are widely spread in the marine environment. PMID- 19191874 TI - Identification of Streptococcus suis genes preferentially expressed under iron starvation by selective capture of transcribed sequences. AB - Streptococcus suis serotype 2 is a zoonotic Gram-positive bacterium responsible for arthritis, meningitis, pneumonia and septicemia in swine and humans. Little information about the regulation of iron on its gene expression had been reported. In this study, 63 S. suis genes upregulated under an iron-restricted condition were identified using selective capture of transcribed sequences (SCUTS) technique: 23 genes involved in metabolism, 22 genes responsible for the replication and genetic information proceeding of the bacteria, eight genes relative to the construction of the cell wall, five ATP-binding cassette transporters, four transcriptional regulators and one uncharacterized gene conserved among streptococcal species. To adapt to the stress, S. suis modulated its physiological activities, which were validated by the upregulation of RelA (a crucial enzyme in stringent response), ArcA (a component of the arginine deiminase system catalyzing the conversion of arginine to ornithine) and CpdB (a cell surface protein that is a substrate of sortase A). All of them were reported to be virulence factors in S. suis or other bacteria. Besides, together with the results of quantitative reverse transcriptase PCR, we found that several homologous genes (fur, fhuGBDA operons) associated with iron uptake as reported in other bacteria were also upregulated under an iron-restricted condition in S. suis. PMID- 19191875 TI - Impaired synthesis and secretion of SopA in Salmonella Typhimurium dam mutants. AB - DNA adenine methylation regulates virulence gene expression in certain bacteria, including Salmonella Typhimurium. The aim of this study was to investigate the involvement of DNA adenine methylase (Dam) methylation in the expression and secretion of the SPI-1 effector protein SopA. For this purpose, SopA-FLAG-tagged wild-type and dam strains of Salmonella Typhimurium were constructed. The expression and secretion of SopA were determined in bacterial culture and in intracellular bacteria recovered from infected HEp-2 epithelial cells. Bacterial culture supernatants and pellets were used to investigate secreted proteins and cell-associated proteins, respectively. Western blot and quantitative reverse transcriptase PCR analysis showed that the dam mutant expresses lower levels of SopA than the wild-type strain. Interestingly, the strain lacking Dam synthesizes SopA under nonpermissive conditions (28 degrees C). In addition, SopA secretion was drastically impaired in the dam mutant. In vivo experiments showed that the intracellular Salmonella dam mutant synthesizes SopA although in lower amounts than the wild-type strain. Taken together, our results suggest that Dam methylation modulates the expression and secretion of SopA in Salmonella Typhimurium. PMID- 19191876 TI - TNF-alpha reduces the level of Staphylococcus epidermidis internalization by bovine endothelial cells. AB - Staphylococcus epidermidis is an environmental opportunistic pathogen associated with bovine intramammary infections. In bacterial infections, the endothelial tissue plays an important role during inflammation and it is the target of proinflammatory cytokines such as tumor necrosis factor alpha (TNF-alpha). Therefore, this work was designed to explore the effect of TNF-alpha on the interaction of S. epidermidis with bovine endothelial cells (BEC). We show that cell signaling activated by TNF-alpha caused a marked reduction in the number of intracellular S. epidermidis, suggesting that molecules participating in this pathway were involved in the internalization of this bacterium. We also found that S. epidermidis internalization was not associated with basal levels of nuclear factor kappa B (NF-kappaB) activity because the intracellular number of bacteria recovered after treating BEC with the NF-kappaB inhibitors, SN50 or BAY 11-7083, was similar to that of the untreated control. Interestingly, inhibition of the basal activity of JNK with SP600125 and p38 with SB203580 caused a decrease in the number of intracellular S. epidermidis. These results suggest that activation of the signaling pathway initiated by TNF-alpha could play an important role in the phagocytosis of this bacterium. However, the basal activity of NF-kappaB was shown not to be important for the internalization process of S. epidermidis. PMID- 19191877 TI - The GacS-GacA two-component regulatory system of Pseudomonas fluorescens: a bacterial two-hybrid analysis. AB - The two-component regulatory system comprised of the sensor kinase, GacS, and its response regulator, GacA, is involved in regulation of secondary metabolism and many other aspects of bacterial physiology. Although it is known that the sensor kinases RetS and LadS feed into the GacS/GacA system, the mechanism through which this occurs is unknown, as are the protein-protein interactions in this system. To characterize and define these interactions, we utilized a bacterial two-hybrid system to study the interactions of GacS and GacA from Pseudomonas fluorescens CHA0. Domains of GacA and GacS, identified through bioinformatics, were subcloned and their ability to interact in vivo was investigated. We found that the entire GacA molecule is required for GacA to interact with itself or GacS. Furthermore, the HisKA/HATPase/REC domains of GacS together are responsible for GacS interacting with GacA, while the HAMP domain of GacS is responsible for GacS interacting with itself. In addition, homologs of Pseudomonas aeruginosa hybrid sensor kinases, RetS and LadS, were identified in P. fluorescens, and shown to interact with GacS, but not GacA. PMID- 19191880 TI - RNA interference in hippocampus demonstrates opposing roles for CREB and PP1alpha in contextual and temporal long-term memory. AB - We injected small interfering RNAs (siRNAs) directly into the hippocampus of wild type mice, knocking down expression of cyclic AMP responsive element-binding protein (CREB) and disrupting long-term, but not short-term, memory after both contextual and trace fear conditioning. In contrast, similar knockdown of siRNA for protein phosphatase 1 (PP1) was sufficient to enhance contextual and temporal memory formation, thereby demonstrating with such a gain-of-function effect a lack of any general deleterious effect for this method of RNAi-mediated gene knockdown. Our findings clearly confirm that contextual memory formation involves CREB and PP1 as positive and negative regulators, respectively, and show for the first time that temporal memory formation shares this mechanism. More generally, we establish that direct injection of siRNA into identified adult brain regions yields specific gene knockdowns, which can be used to validate in vivo candidate genes involved in behavioral plasticity. PMID- 19191881 TI - Joint sustainability of petroleum energy production and water resources. PMID- 19191878 TI - Genetic modulation of striatal volume by loci on Chrs 6 and 17 in BXD recombinant inbred mice. AB - Natural variation in the absolute and relative size of different parts of the human brain is substantial, with a range that often exceeds a factor of 2. Much of this variation is generated by the cumulative effects of sets of unknown gene variants that modulate the proliferation, growth and death of neurons and glial cells. Discovering and testing the functions of these genes should contribute significantly to our understanding of differences in brain development, behavior and disease susceptibility. We have exploited a large population of genetically well-characterized strains of mice (BXD recombinant inbred strains) to map gene variants that influence the volume of the dorsal striatum (caudate-putamen without nucleus accumbens). We used unbiased methods to estimate volumes bilaterally in a sex-balanced sample taken from the Mouse Brain Library (www.mbl.org). We generated a matched microarray data set to efficiently evaluate candidate genes (www.genenetwork.org). As in humans, volume of the striatum is highly heritable, with greater than twofold differences among strains. We mapped a locus that modulates striatal volume on chromosome (Chr) 6 at 88 +/- 5 Mb. We also uncovered an epistatic interaction between loci on Chr 6 and Chr 17 that modulates striatal volume. Using bioinformatic tools and the corresponding expression database, we have identified positional candidates in these quantitative trait locus intervals. PMID- 19191883 TI - Ground water development--the time to full capture problem. AB - Ground water systems can be categorized with respect to quantity into two groups: (1) those that will ultimately reach a new equilibrium state where pumping can be continued indefinitely and (2) those in which the stress is so large that a new equilibrium is impossible; hence, the system has a finite life. Large ground water systems, where a new equilibrium can be reached and in which the pumping is a long distance from boundaries where capture can occur, take long times to reach a new equilibrium. Some systems are so large that the new equilibrium will take a millennium or more to reach a new steady-state condition. These large systems pose a challenge to the water manager, especially when the water manager is committed to attempting to reach a new equilibrium state in which water levels will stabilize and the system can be maintained indefinitely. PMID- 19191884 TI - Recollections of an inquiring engineer. PMID- 19191885 TI - Distinguishing iron-reducing from sulfate-reducing conditions. AB - Ground water systems dominated by iron- or sulfate-reducing conditions may be distinguished by observing concentrations of dissolved iron (Fe(2+)) and sulfide (sum of H(2)S, HS(-), and S(=) species and denoted here as "H(2)S"). This approach is based on the observation that concentrations of Fe(2+) and H(2)S in ground water systems tend to be inversely related according to a hyperbolic function. That is, when Fe(2+) concentrations are high, H(2)S concentrations tend to be low and vice versa. This relation partly reflects the rapid reaction kinetics of Fe(2+) with H(2)S to produce relatively insoluble ferrous sulfides (FeS). This relation also reflects competition for organic substrates between the iron- and the sulfate-reducing microorganisms that catalyze the production of Fe(2+) and H(2)S. These solubility and microbial constraints operate in tandem, resulting in the observed hyperbolic relation between Fe(2+) and H(2)S concentrations. Concentrations of redox indicators, including dissolved hydrogen (H(2)) measured in a shallow aquifer in Hanahan, South Carolina, suggest that if the Fe(2+)/H(2)S mass ratio (units of mg/L) exceeded 10, the screened interval being tapped was consistently iron reducing (H(2) approximately 0.2 to 0.8 nM). Conversely, if the Fe(2+)/H(2)S ratio was less than 0.30, consistent sulfate reducing (H(2) approximately 1 to 5 nM) conditions were observed over time. Concomitantly high Fe(2+) and H(2)S concentrations were associated with H(2) concentrations that varied between 0.2 and 5.0 nM over time, suggesting mixing of water from adjacent iron- and sulfate-reducing zones or concomitant iron and sulfate reduction under nonelectron donor-limited conditions. These observations suggest that Fe(2+)/H(2)S mass ratios may provide useful information concerning the occurrence and distribution of iron and sulfate reduction in ground water systems. PMID- 19191886 TI - Impact of sea-level rise on sea water intrusion in coastal aquifers. AB - Despite its purported importance, previous studies of the influence of sea-level rise on coastal aquifers have focused on specific sites, and a generalized systematic analysis of the general case of the sea water intrusion response to sea-level rise has not been reported. In this study, a simple conceptual framework is used to provide a first-order assessment of sea water intrusion changes in coastal unconfined aquifers in response to sea-level rise. Two conceptual models are tested: (1) flux-controlled systems, in which ground water discharge to the sea is persistent despite changes in sea level, and (2) head controlled systems, whereby ground water abstractions or surface features maintain the head condition in the aquifer despite sea-level changes. The conceptualization assumes steady-state conditions, a sharp interface sea water fresh water transition zone, homogeneous and isotropic aquifer properties, and constant recharge. In the case of constant flux conditions, the upper limit for sea water intrusion due to sea-level rise (up to 1.5 m is tested) is no greater than 50 m for typical values of recharge, hydraulic conductivity, and aquifer depth. This is in striking contrast to the constant head cases, in which the magnitude of salt water toe migration is on the order of hundreds of meters to several kilometers for the same sea-level rise. This study has highlighted the importance of inland boundary conditions on the sea-level rise impact. It identifies combinations of hydrogeologic parameters that control whether large or small salt water toe migration will occur for any given change in a hydrogeologic variable. PMID- 19191888 TI - Pain in the numb chin syndrome. PMID- 19191889 TI - The interaction between Helicobacter pylori and atopy: does inverse association really exist? AB - AIM: To date, cross-sectional and case-control studies suggest an inverse association between Helicobacter pylori infection and atopic diseases, whereas the immunologic basis has not been studied yet. In this study we investigated T helper (Th) cell function in H. pylori-infected children and compared cytokine responses in atopic and non-atopic groups. METHODS: The study groups was recruited from a cohort of 327 healthy children evaluated and followed-up for 6 years to assess the natural history of H. pylori infection. Seventy-four of 136 healthy children who underwent (13)C urea breath test were eligible and accepted to participate. All participants were evaluated by a questionnaire, and skin prick testing. According to the results, children were divided into four groups with respect to the presence or absence of H. pylori and atopy. Peripheral blood mononuclear cells isolated from 34 of 74 children were cultured with H. pylori, Der p 1, and phytohemagglutinin (PHA). Interferon-gamma (IFN-gamma), interleukin (IL)-4 and IL-10, transforming growth factor-beta (TGF-beta) levels were measured in supernatants. RESULTS: The frequency of atopy was lower in H. pylori-infected group (31.9% vs. 48.1, p = .22), while atopic symptoms were similar between infected and non-infected children. While PHA and H. pylori induced IFN-gamma levels were significantly higher in H. pylori-infected children, concomitant presence of both atopy and H. pylori decreased the level of PHA and H. pylori induced IFN-gamma production. PHA and Der p 1-induced IL-4 levels were higher in atopic children, and IL-4 production was suppressed when they were concomitantly infected with H. pylori. The production of TGF-beta was found to be suppressed in atopic children irrespective of the presence of H. pylori infection. CONCLUSION: The results of the current study demonstrated a counteractive Th1 and Th2 cytokine interaction between H. pylori infection and atopy. However, this counteractive immunologic balance did not protect against atopy. PMID- 19191890 TI - Autoimmune angioneurotic edema in a patient with Helicobacter pylori infection. AB - Association of acquired autoimmune angioneurotic edema with other diseases is increasing. However, the precise mechanism by which antibodies to C1-esterase inhibitor (C1-INH) are produced, is not elucidated. We describe a patient with IgA antibodies against C1-INH without other autoimmune markers. Our patient had gastritis and Helicobacter pylori infection, proven by biopsy. This case suggests that H. pylori infection can act as triggering factor for acquired autoimmune angioneurotic edema. PMID- 19191891 TI - Comparison of high resolution magnifying endoscopy and standard videoendoscopy for the diagnosis of Helicobacter pylori gastritis in routine clinical practice: a prospective study. AB - BACKGROUND: It has been shown that standard endoscopic features often labeled as gastritis has a poor correlation with histopathology. Recently, high resolution magnifying endoscopy has been reported to be an effective method to diagnose gastritis. The aim of the present study was to compare standard endoscopy with magnifying endoscopy for the diagnosis of Helicobacter pylori gastritis, and to determine whether gastritis can be diagnosed based on findings at magnification endoscopy. MATERIALS AND METHODS: A total of 129 patients were enrolled into the study. Erythema, erosions, prominent area gastrica, nodularity, and regular arrangement of collecting venules (RAC) were investigated by standard endoscopy. Standard endoscopy was followed by magnifying endoscopy in all patients, and repeated in 55 patients after indigo carmine spraying. RESULTS: None of the standard endoscopic features showed a sensitivity of more than 70% for H. pylori gastritis, except RAC pattern analysis. Absence of a corporal RAC pattern had 85.7% sensitivity and 82.8% specificity for predicting H. pylori infection. Under magnification, the sensitivity and specificity of regular corporal pattern (regular collecting and capillary vascular structures with gastric pits resembling pinholes) for predicting normal histology were 90.3% and 93.9%, respectively. Loss of collecting venules, or both collecting and capillary structures was correlated with chronic inflammation and activity. With the progression of mucosal atrophy, irregular collecting venules became visible. The values for irregularly arranged antral ridge pattern for the prediction of antral gastritis were 89.3% and 65.2%, respectively. Indigo carmine staining increased sensitivity and specificity up to 97.6% and 100% for corporal gastritis, and up to 88.4% and 75.0% for antral gastritis, respectively. Indigo carmine staining significantly increases the detection of intestinal metaplasia. CONCLUSIONS: High resolution magnifying is superior to standard endoscopy for the diagnosis of H. pylori gastritis, and identification of specific histopathologic features such as atrophy and intestinal metaplasia seems possible. PMID- 19191892 TI - Dual-priming oligonucleotide-based multiplex PCR for the detection of Helicobacter pylori and determination of clarithromycin resistance with gastric biopsy specimens. AB - BACKGROUND: Assessment of Helicobacter pylori (H. pylori) clarithromycin resistance has rarely been performed routinely despite an increasing resistance rate. Our aim was to develop and evaluate the use of dual-priming oligonucleotide (DPO)-based multiplex polymerase chain reaction (PCR) to detect point mutations in the 23S rRNA gene responsible for clarithromycin resistance of H. pylori. MATERIALS AND METHODS: Gastric biopsy specimens from 212 untreated patients with dyspepsia were examined by culture, histology, and DPO-based multiplex PCR. A disk diffusion test and E-test were used for performing phenotypic antibiotic susceptibility tests. RESULTS: Among the biopsy specimens tested, 22.2% (47/212), 42.5% (90/212), and 41.5% (88/212) of the specimens were classified as H. pylori positive by culture, histology, and DPO-based multiplex PCR, respectively. Among 96 strains identified by either culture or DPO-based multiplex PCR, 80 strains were clarithromycin-susceptible and 16 strains (16.7%) were clarithromycin resistant. There was 94.1% (32/34) concordance between phenotypic susceptibility tests and DPO-based multiplex PCR. In two patients with discrepant results, only DPO-based multiplex PCR detected clarithromycin-resistant strains. DPO-based multiplex PCR identified additional 49 clarithromycin-resistant or clarithromycin susceptible H. pylori among 165 culture-negative specimens. CONCLUSIONS: DPO based multiplex PCR can be used as a practical method for the detection of H. pylori infection and the determination of clarithromycin susceptibility in addition to phenotypic antimicrobial susceptibility tests. PMID- 19191893 TI - Helicobacter pylori eradication therapy success regarding different treatment period based on clarithromycin or metronidazole triple-therapy regimens. AB - BACKGROUND: The study compares the eradication success of standard first-line triple therapies of different durations (7, 10, and 14 days). MATERIALS AND METHODS: A total of 592 naive Helicobacter pylori-positive patients were randomized to receive pantoprazole, amoxicillin, and clarithromycin or metronidazole for 14 days (PACl14 or PAM14), 10 days (PACl10 or PAM10), or 7 days (PACl7 or PAM7). H. pylori eradication was assessed by histological, microbiological, and rapid urease examination. RESULTS: The intention-to-treat (ITT) and per-protocol (PP) analyses have shown no overall statistically significant differences between the eradication success of PACl and PAM treatment groups (ITT p = .308, PP p = .167). Longer treatment duration has yielded statistically significant increase in eradication success for clarithromycin (ITT p = .004; PP p = .004) and metronidazole (ITT p = .010; PP p = .034) based regimens. Namely, PACl10, PACl14, and PAM14 protocols resulted in eradication success exceeding 80% in ITT and 90% in PP analysis. Primary resistance to clarithromycin and metronidazole equals 8.2% and 32.9%, respectively. Prolonging the metronidazole-based treatment duration in patients with resistant strains resulted in statistically significant higher eradication success. CONCLUSIONS: For all antimicrobial combinations, 14 days protocols have led to a significant increase of H. pylori eradication success when compared to 10 and 7 days, respectively. Prolonging the treatment duration can overcome the negative effect of metronidazole resistance. Only PAM14, PACl10 protocols achieved ITT success > 80% and should be recommended as the first line eradication treatment in Croatia. PMID- 19191894 TI - The expression of iron-repressible outer membrane proteins in Helicobacter pylori and its association with iron deficiency anemia. AB - BACKGROUND: Helicobacter pylori infection is known to be a cause of iron deficiency anemia (IDA) that is unresponsive to iron supplements. H. pylori bind iron to a specific receptor by iron-repressible outer membrane proteins (IROMPs) under conditions of restricted iron. MATERIALS AND METHODS: We compared the expression of IROMPs from strains of H. pylori under both iron-restricted and iron-supplemented conditions to determine the difference between strains with and without IDA. One standard strain, two clinical strains, and three IDA strains were cultured; and then the IROMPs were extracted under iron-restricted and iron supplemented conditions. We used SDS-PAGE to compare the expression of the IROMPs from each strain. RESULTS: IROMPs were found in IDA strains under iron-restricted conditions and their molecular sizes were estimated to be 56, 48, 41, and 37 kDa. In the iron-repleted media, the IROMPs were no longer present. CONCLUSION: In the iron-depleted state, specific H. pylori strains associated with IDA demonstrated an advantage in iron acquisition due to a higher expression of IROMPs. Our results can explain in part why some patients with H. pylori infection are more prone to develop clinical IDA under restricted iron conditions in the host. PMID- 19191895 TI - Childhood hygienic practice and family education status determine the prevalence of Helicobacter pylori infection in Iran. AB - BACKGROUND: Management of Helicobacter pylori, a causative agent of gastrointestinal diseases is an important health problem in most countries. The main reasons include poorly defined epidemiological status and unrecognized mode of bacterial transmission. Our objective was to investigate the prevalence of H. pylori infection in a representative population of Iran and to evaluate possible risk factors for the H. pylori infection. MATERIALS AND METHODS: In this cross sectional study, 2561 healthy individuals aged 18-65 years (mean age, 35.5 years) were selected out of 12,100,000 inhabitants of Tehran province by cluster sampling. Infection with H. pylori was evaluated by detection of anti-H. pylori IgG antibody in serum. Sociodemographic status of each subject was determined by filling up a questionnaire. RESULTS: Prevalence of H. pylori infection was 69% and was correlated with increasing age. The highest infection rate (79.2%) was seen in individuals 46-55 years old. No association was detected between H. pylori positivity and gender. Low education of the study subjects; low father's and mother's education; poor tooth brushing habit; crowded families in childhood; and lack of household bath, hygienic drinking water, and swage disposal facility in childhood were determined as possible risk factors. CONCLUSIONS: The rate of prevalence of H. pylori infection was higher than developed countries. Low socioeconomic status, poor sanitary indications, and crowded families in childhood were related to high prevalence of H. pylori infection in Iran. Accordingly, fecal-oral and oral-oral routes could be considered as the main pathways of transmission of H. pylori. PMID- 19191896 TI - Toll-like receptor 4 +3725 G/C polymorphism, Helicobacter pylori seropositivity, and the risk of gastric atrophy and gastric cancer in Japanese. AB - BACKGROUND: Toll-like receptor 4 (TLR4) Asp299Gly and Thr399Ile polymorphisms were reported to be a risk factor of gastric carcinoma or its precursors in Caucasian and Indian population, but these polymorphisms are absent in Japanese. We investigated the associations of TLR4+3725 G/C polymorphism, another functional polymorphism of TLR4, with risk of gastric cancer and gastric atrophy in Japanese. MATERIALS AND METHODS: Study subjects were 583 histologically diagnosed gastric cancer patients and age- and sex-matched 1592 control outpatients, who visited Aichi Cancer Center Hospital from 2001 to 2005. Serum anti-H. pylori IgG antibody and pepsinogens were measured to evaluate H. pylori infection and gastric atrophy, respectively. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated by a logistic model. RESULTS: Among the seropositive subjects, the age- and sex-adjusted OR of gastric atrophy was 1.17 (95%CI: 0.91-1.50) for G/C, 1.20 (95%CI: 0.76-1.89) for C/C, and 1.18 (95%CI: 0.93-1.49) for G/C+C/C relative to G/G genotype. The age- and sex-adjusted OR of severe gastric atrophy among H. pylori seropositive subjects was 1.43 (95%CI: 0.99-2.06) for G/C, 1.47 (95%CI: 0.76-2.88) for C/C, and 1.43 (95%CI: 1.01-2.04) for G/C+C/C. The OR of gastric cancer compared with gastric atrophy controls was not statistically significant. CONCLUSION: Our study found that TLR4+3725 G/C polymorphism was a risk factor of severe gastric atrophy in H. pylori seropositive Japanese. Our results underscored the significance of the variations in host innate immunity due to TLR4 polymorphism as genetic predispositions to gastric precancerous lesions in Eastern Asian populations with the same backgrounds. PMID- 19191897 TI - Gender difference of circulating ghrelin and leptin concentrations in chronic Helicobacter pylori infection. AB - BACKGROUND: Both ghrelin and leptin are important appetite hormones secreted from the stomach. We examined whether demographic background, Helicobacter pylori infection, or its related gastritis severity could be associated with circulating ghrelin and leptin levels. METHODS: This study prospectively enrolled 341 dyspeptic patients (196 females, 145 males), who had received endoscopy to provide the gastric specimens over both antrum and corpus for histology reviewed by the updated Sydney's system. The fasting blood sample of each patient was obtained for total ghrelin and leptin analysis. RESULTS: Without H. pylori infection, there were similar ghrelin levels between female and male patients. In the H. pylori-infected patients, the males had lower plasma ghrelin levels than females (1053 vs. 1419 pg/mL, p < .001). Only in males, not in females, the H. pylori infection and its related acute and chronic inflammation scores were significantly associated with a lower ghrelin level (p < or = .04). The multivariate regression disclosed that only the chronic inflammation score independently related to a lower ghrelin level. Only in males, the ghrelin levels ranked in a downward trend for the gastritis feature as with limited-gastritis, with antrum-predominant gastritis, and with corpus-gastritis (1236, 1101, and 977 pg/mL). Leptin level was not related to H. pylori-related gastritis, but positively related to body mass index. CONCLUSION: There should be a gender difference to circulating total ghrelin levels, but not leptin levels, in response to H. pylori infection and its related chronic gastritis. PMID- 19191899 TI - Characterization and application of a new monoclonal antibody with high specificity for Helicobacter hepaticus. AB - BACKGROUND AND AIMS: Infection with Helicobacter hepaticus is suggested to play a role in the pathogenesis of chronic liver disease in humans. However, reactive antigens among Helicobacter species make the development of an H. hepaticus ELISA test with high specificity difficult. A new monoclonal antibody from a hybridoma clone (HRII-51) showed high specificity to H. hepaticus without cross-reaction to other gastrointestinal bacteria. METHODS: The molecular weight of HRII-51 immunoreactive antigen was examined by Western blot of H. hepaticus probed with the monoclonal antibody HRII-51. A HRII-51-immunoreactive antigen capture ELISA was prepared in which the specific antigen was anchored by HRII-51-immobilized ELISA plate. Accuracy of HRII-51 antigen capture ELISA was examined using sera obtained from mice inoculated with Helicobacter species. Specificity of HRII-51 antigen capture ELISA was compared to that of H. hepaticus antigen-based ELISA using human sera with absorption by H. pylori cell lysate. RESULTS: HRII-51 immunoreactive antigen had a molecular weight of 15 kDa. Sensitivity and specificity of HRII-51 antigen capture ELISA were 87.0% and 97.6% in mice inoculated with Helicobacter species. In human sera, modification of the results by absorption with H. pylori lysate was smaller in HRII-51 antigen capture ELISA comparing with H. hepaticus-antigen-based ELISA. CONCLUSION: Use of the HRII-51 antigen capture ELISA would be a useful approach for the serodiagnosis of H. hepaticus infection in both experimental animals and humans. PMID- 19191898 TI - Prevalence of primary fluoroquinolone resistance among clinical isolates of Helicobacter pylori at a University Hospital in Southern Taiwan. AB - BACKGROUND: Fluoroquinolone-containing therapy is effective in eradicating Helicobacter pylori. However, the resistance rate of H. pylori to fluoroquinolones in Taiwan has not yet been reported. In this study, we aimed to investigate the susceptibility to antibiotics commonly used in eradication schedules and fluoroquinolones in H. pylori. METHODS: A total of 210 clinical isolates of H. pylori were collected from April 1998 to September 2007 from patients in southern Taiwan. The in vitro activities of six antimicrobial agents were determined by the agar dilution method and Etest. The mutations in quinolone resistance-determining regions of gyrA and gyrB were investigated by direct sequencing. RESULTS: Overall, 5.7% of the isolates were resistant to ciprofloxacin and levofloxacin. The resistance rate to amoxicillin, clarithromycin, metronidazole, and tetracycline was 1.0% (two of 210), 9.5% (20 of 210), 27.6% (58 of 210), and 0.5% (one of 210), respectively. The resistance rate to either ciprofloxacin or to levofloxacin increased from 2.8% (1998-2003) to 11.8% (2004-2007). The mutations in gyrA at N87 or D91 had an impact on primary fluoroquinolone resistance in H. pylori. Garenoxacin, but not moxifloxacin, had a good in vitro inhibitory effect against ciprofloxacin/levofloxacin-resistant strains compared with objective minimal inhibitory concentration values. CONCLUSIONS: Drug resistance to ciprofloxacin and levofloxacin in H. pylori collected from 2004 to 2007 increased significantly compared with resistance level observed during 1998-2003. The continuous surveillance of quinolone resistance among H. pylori is important in this area. PMID- 19191900 TI - The Helicobacter hepaticus hefA gene is involved in resistance to amoxicillin. AB - BACKGROUND: Gastrointestinal infections with pathogenic Helicobacter species are commonly treated with combination therapies, which often include amoxicillin. Although this treatment is effective for eradication of Helicobacter pylori, the few existing reports are less clear about antibiotic susceptibility of other Helicobacter species. In this study we have determined the susceptibility of gastric and enterohepatic Helicobacter species to amoxicillin, and have investigated the mechanism of amoxicillin resistance in Helicobacter hepaticus. MATERIALS AND METHODS: The minimal inhibitory concentration (MIC) of antimicrobial compounds was determined by E-test and agar/broth dilution assays. The hefA gene of H. hepaticus was inactivated by insertion of a chloramphenicol resistance gene. Transcription was measured by quantitative real-time polymerase chain reaction. RESULTS: Three gastric Helicobacter species (H. pylori, H. mustelae, and H. acinonychis) were susceptible to amoxicillin (MIC < 0.25 mg/L). In contrast, three enterohepatic Helicobacter species (H. rappini, H. bilis, and H. hepaticus) were resistant to amoxicillin (MIC of 8, 16, and 6-64 mg/L, respectively). There was no detectable beta-lactamase activity in H. hepaticus, and inhibition of beta-lactamases did not change the MIC of amoxicillin of H. hepaticus. A H. hepaticus hefA (hh0224) mutant, encoding a TolC-component of a putative efflux system, resulted in loss of amoxicillin resistance (MIC 0.25 mg/L), and also resulted in increased sensitivity to bile acids. Finally, transcription of the hefA gene was not responsive to amoxicillin, but induced by bile acids. CONCLUSIONS: Rodents are frequently colonized by a variety of enterohepatic Helicobacter species, and this may affect their global health status and intestinal inflammatory responses. Animal facilities should have treatment strategies for Helicobacter infections, and hence resistance of enterohepatic Helicobacter species to amoxicillin should be considered when designing eradication programs. PMID- 19191901 TI - High alpha-defensin levels in patients with systemic lupus erythematosus. AB - Innate immunity plays a role in systemic lupus erythematosus (SLE). Our objective was to determine the levels of defensins, which are antimicrobial and immunomodulatory polypeptides, in SLE. Sera from SLE patients and healthy controls were tested for pro-inflammatory human beta-defensin 2 (hBD-2) and for alpha-defensin human neutrophil peptide 1 (HNP-1). hBD-2 could not be detected by enzyme-linked immunosorbent assay (ELISA) and its mRNA levels were low in SLE patients and similar to those found in controls. In contrast, the mean alpha defensin level in the sera of all SLE patients (11.07 +/- 13.92 ng/microl) was significantly higher than that of controls (0.12 +/- 0.07 ng/microl). Moreover, 60% of patients demonstrated very high serum levels (18.5 +/- 13.36 ng/microl) and 50% showed elevated gene expression in polymorphonuclear cells. High alpha defensin levels correlated with disease activity, but not with neutrophil count. Thus, activation and degranulation of neutrophils led to alpha-defensin secretion in SLE patients. Given the immunomodulatory role of alpha-defensins, it is possible that their secretion may activate the adaptive immune system leading to a systemic response. PMID- 19191902 TI - Suppression of allergic airway inflammation in a mouse model by Der p2 recombined BCG. AB - Allergic asthma is a chronic inflammatory disease mediated by T helper (Th)2 cell immune responses. Currently, immunotherapies based on both immune deviation and immune suppression, including the development of recombinant mycobacteria as immunoregulatory vaccines, are attractive treatment strategies for asthma. In our previous studies, we created a genetically recombinant form of bacille Calmette Guerin (rBCG) that expressed Der p2 of house dust mites and established that it induced a shift from a Th2 response to a Th1 response in naive mice. However, it is unclear whether rBCG could suppress allergic airway inflammation in a mouse model. In this article we report that rBCG dramatically inhibited airway inflammation, eosinophilia, mucus production and mast cell degranulation in allergic mice. Analysis of interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) levels in bronchoalveolar lavage fluid (BALF) and lung tissue revealed that the suppression was associated with a shift from a Th2 response to a Th1 response. At the same time, rBCG induced a CD4(+) CD25(+) Foxp3(+) T-cell subtype that could suppress the proliferation of Th2 effector cells in vitro in an antigen-specific manner. Moreover, suppression of CD4(+) CD25(+) T cells could be adoptively transferred. Thus, our results demonstrate that rBCG induces both generic and specific immune responses. The generic immune response is associated with a shift from a Th2 to a Th1 cytokine response, whereas the specific immune response against Der p2 appears to be related to the expansion of transforming growth factor-beta (TGF-beta)-producing CD4(+) CD25(+) Foxp3(+) regulatory T cells. rBCG can suppress asthmatic airway inflammation through both immune deviation and immune suppression and may be a feasible, efficient immunotherapy for asthma. PMID- 19191903 TI - Regulation of different inflammatory diseases by impacting the mevalonate pathway. AB - The 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase inhibitors (statins) interfere with the mevalonate pathway. While initially developed for their lipid lowering properties, statins have been extensively investigated with respect to their impact on autoantigen and alloantigen driven immune responses. Mechanistically it was shown that statins modify immune responses on several levels, including effects on dendritic cells, endothelial cells, macrophages, B cells and T cells. Several lines of evidence suggest that statins act in a disease-specific manner and are not effective in each immune disorder. This review discusses possible modes of action of statins in modulating immunity towards autoantigens and alloantigens. PMID- 19191904 TI - Differential gene expression of peripheral blood mononuclear cells from rheumatoid arthritis patients may discriminate immunogenetic, pathogenic and treatment features. AB - This study aimed to evaluate the association between the differential gene expression profiling of peripheral blood mononuclear cells of rheumatoid arthritis patients with their immunogenetic (human leucocyte antigen shared epitope, HLA-SE), autoimmune response [anti-cyclic citrullinated peptide (CCP) antibodies], disease activity score (DAS-28) and treatment (disease-modifying antirheumatic drugs and tumour necrosis factor blocker) features. Total RNA samples were copied into Cy3-labelled complementary DNA probes, hybridized onto a glass slide microarray containing 4500 human IMAGE complementary DNA target sequences. The Cy3-monocolour microarray images from patients were quantified and normalized. Analysis of the data using the significance analysis of microarrays algorithm together with a Venn diagram allowed the identification of shared and of exclusively modulated genes, according to patient features. Thirteen genes were exclusively associated with the presence of HLA-SE alleles, whose major biological function was related to signal transduction, phosphorylation and apoptosis. Ninety-one genes were associated with disease activity, being involved in signal transduction, apoptosis, response to stress and DNA damage. One hundred and one genes were associated with the presence of anti-CCP antibodies, being involved in signal transduction, cell proliferation and apoptosis. Twenty-eight genes were associated with tumour necrosis factor blocker treatment, being involved in intracellular signalling cascade, phosphorylation and protein transport. Some of these genes had been previously associated with rheumatoid arthritis pathogenesis, whereas others were unveiled for future research. PMID- 19191905 TI - Apolipoproteins inhibit the innate immunity activated by necrotic cells or bacterial endotoxin. AB - We suggested earlier that the hydrophobic portions (Hyppos) of molecules, which are normally embedded in the membranes of cells or the core of molecular structures so as to be separated from the aqueous environment, might serve as evolutionarily ancient alarm signals of injury or stress to initiate innate immune responses when they are exposed on the surface. Under normal physiological conditions, the Hyppos released from endogenous or exogenous sources might be handled by 'Hyppo-quenchers'in vivo to maintain the tissue homeostasis and immune modulation. To test this idea, we selected apolipoproteins, which have been known to transport blood lipids and play a role in a number of pathological inflammatory conditions. We examined their role as Hyppo-quenchers in early immune responses and found that apolipoproteins showed significant inhibition of the nuclear factor-kappaB-dependent gene expression in recombinant Chinese hamster ovary (CHO) cells and dendritic cells stimulated by necrotic cells or bacterial endotoxin. In addition, our results indicate that apolipoproteins could dramatically abrogate complement fixation on the surface of necrotic cells. These findings suggest that apolipoproteins, besides having known functions in lipid metabolism, also have a role in preventing the initiation of innate immunity, potentially through neutralizing Hyppos from injured cells or exogenous endotoxin. PMID- 19191906 TI - Cytotoxic T-lymphocyte antigen 4 blockade augments the T-cell response primed by attenuated Listeria monocytogenes resulting in more rapid clearance of virulent bacterial challenge. AB - Cytotoxic T-lymphocyte antigen 4 (CTLA-4) uniformly suppresses antigen-specific T cells during chronic infection with bacterial, parasitic or viral pathogens. However, the importance of CTLA-4 in controlling the T-cell response during acute infection or after priming with live attenuated vaccine vectors has not been well characterized. Since strategies aimed at blocking CTLA-4 are being actively developed to therapeutically augment T-cell-mediated immunity, the effects of CTLA-4 blockade on T-cell activation during these conditions need to be more clearly defined. We have examined the role of CTLA-4 in a prime-challenge model of acute bacterial infection using both attenuated and virulent strains of the intracellular bacterium Listeria monocytogenes. Although Foxp3(+) CD4(+) T cells are the predominant CTLA-4-expressing cell type in naive mice, antigen-specific Foxp3(-) CD4(+) cells upregulate CTLA-4 expression after primary L. monocytogenes infection. Blockade of CTLA-4 results in increased numbers of L. monocytogenes specific CD4 and CD8 T cells after primary infection with attenuated L. monocytogenes, and confers more rapid bacterial clearance after secondary challenge with virulent L. monocytogenes. Accordingly, CTLA-4 plays an important suppressive role in T-cell priming and protective immunity in a prime-challenge model of acute bacterial infection. PMID- 19191907 TI - Calpain inhibition induces activation of the distinct signalling pathways and cell migration in human monocytes. AB - We have recently reported that constitutively active calpain negatively regulates activation of the distinct signalling pathways and cell migration in human neutrophils. Here, we report that a similar regulatory system is also functioning in human monocytes, but not lymphocytes. Calpain was constitutively active in resting human monocytes, but not lymphocytes. Mitogen-activated protein kinases, including extracellular signal-regulated kinase (ERK), p38 and c-Jun N-terminal kinase (JNK), phosphatidylinositol 3-kinase (PI3K)/Akt and p21-activated kinase (PAK, an effector molecule of Rac) were rapidly (within 1 min) activated in monocytes, but not lymphocytes, upon exposure to calpain inhibitors (PD150606 and N-acetyl-Leu-Leu-Nle-CHO), but not PD145305 (the inactive analogue of PD150606). Following activation of these signalling pathways, monocytes displayed active migration within 5 min after exposure to calpain inhibitors, and active migration was sustained for more than 45 min. The micropipette method revealed that calpain inhibition-mediated monocyte migration was chemotaxis, not random migration. The studies with pharmacological inhibitors suggest that calpain inhibition-mediated monocyte migration is mediated by activation of ERK, p38, JNK, PI3K/Akt and Rac. NSC23766 (Rac inhibitor) and pertussis toxin (PTX) suppressed calpain inhibitor induced phosphorylation of distinct signalling molecules (PAK, ERK, p38, JNK and Akt) as well as cell migration, suggesting that the PTX-sensitive G protein and Rac axis may be a possible key target of calpain inhibitors. These findings suggest that constitutively active calpain negatively regulates activation of the distinct signalling pathways and cell migration in resting monocytes, but not lymphocytes. PMID- 19191908 TI - Role of high-affinity HLA-DP specific CLIP-derived peptides in beryllium binding to the HLA-DPGlu69 berylliosis-associated molecules and presentation to beryllium sensitized T cells. AB - Berylliosis is driven by the accumulation in the lung of beryllium-specific T helper type 1 (Th1) cells recognizing beryllium as antigen when presented principally by human leucocyte antigen DP molecules carrying a glutamate at position beta69 (HLA-DPGlu69). This study was designed to clarify the precise role of peptides in beryllium binding to the HLA-DP groove's pocket 4 and to identify peptides with higher affinity for pocket 4 that might prevent beryllium presentation and T-cell stimulation. Beryllium/HLA-DP interactions were analysed by the ability of beryllium to compete with CLIP and CLIP-derived peptides to HLA DPGlu69 soluble molecule. The CLIP-derived low-affinity peptide CLIP-AA, could not outcompete beryllium; while the CLIP-derived high-affinity peptides CLIP-YY, CLIP-QY and CLIP-RF were only marginally influenced by the presence of beryllium in the competition assay. The effect of these CLIP-derived high-affinity peptides on beryllium presentation was determined by measuring interferon-gamma (IFN gamma) release upon beryllium stimulation of peripheral blood mononuclear cells obtained from beryllium-hypersensitive subjects. CLIP-YY did inhibit beryllium presentation and T-cell activation, while CLIP-QY and CLIP-RF markedly enhanced the IFN-gamma response to beryllium. Anti-HLA-DP monoclonal antibody blocked the beryllium-induced IFN-gamma release in the presence of CLIP-QY (88%) and CLIP-RF (76%). A similar effect was observed for CLIP-YY capability to block IFN-gamma release by beryllium stimulation in the presence of CLIP-QY (79%) and CLIP-RF (76%). Overall, these data support the proposal that HLA-DP high-affinity peptides might be used as a model for specific berylliosis therapy. PMID- 19191909 TI - Effect of bone morphogenetic protein-6 on macrophages. AB - Bone morphogenetic proteins (BMPs) are members of the transforming growth factor (TGF)-beta superfamily which regulates bone formation, haematopoiesis and development. While TGF-beta is known to be a negative regulator of the immune system, the effect of BMPs on the immune system is largely unknown. Herein, the effect of BMP-6 on the innate immune system was investigated using the murine macrophage cell line RAW 264.7. BMP-6 altered cellular morphology, inhibited cellular proliferation, increased the fraction of subG(1) phase cells, and decreased the fraction of cells in the S and G(2)M phases, without changing the percentage of apoptotic cells. In addition, BMP-6 induced expression of pro inflammatory inducible nitric oxide synthase (iNOS) and the cytokine tumour necrosis factor (TNF)-alpha. Reverse transcription-polymerase chain reaction (RT PCR) analysis demonstrated the expression of all three known type II BMP receptors [BMP-RII, activin (Act)-RIIA and Act-RIIB] and two of the three known type I receptors [activin receptor-like kinase 2 (ALK2) and ALK3]. Over expression as well as knock-down studies using short hairpin RNA (shRNA) demonstrated that BMP-RII, ALK2 and ALK3 are the functional BMP-6 receptors in macrophages. Finally, the effect of BMP-6 was confirmed in murine peritoneal macrophages and the THP-1 human monocyte cell line. Taken together, these results demonstrate that BMP-6 regulates the proliferation and gene expression profile of macrophages. PMID- 19191911 TI - Interleukin-4 suppresses the cytotoxic potential of in vitro generated, adaptive regulatory CD4 T cells by down-regulation of granzyme B. AB - Regulatory CD4+ T cells (Tregs) control immune responses using secretion of anti inflammatory cytokines and/or cytotoxic mechanisms and play a central role in the outcomes of several immune pathologies. Previous studies suggest an impaired function of Tregs in allergy, especially during allergen seasons, but the underlying mechanism is not known. Therefore, we analysed the impact of the T helper type 2 cytokine interleukin (IL)-4 on in vitro generated adaptive Tregs (aTregs), which have been reported to use the granzyme B (GrB)/perforin pathway to kill autologous immune cells. aTregs were generated by co-ligation of CD3 and CD46 on CD4+ T lymphocytes and granzyme expression was analysed using flow cytometry. To quantify GrB and perforin expression as well as IL-10 secretion in response to IL-4, specific enzyme-linked immunosorbent assays were performed in cell lysates and/or culture supernatants. Using a flow cytometry-based cytotoxicity assay the impact of IL-4 on the cytotoxic potential of aTregs was investigated. While IL-4 did not affect IL-10 secretion and perforin expression in aTregs, a significant suppression of GrB synthesis was detected in the presence of IL-4. In addition, IL-4-mediated suppression of GrB led to impaired cytotoxicity of aTregs against K562 target cells. In conclusion, our data suggest that IL-4 might play a role in impaired aTreg function in allergy. PMID- 19191910 TI - Evolution of the opossum major histocompatibility complex: evidence for diverse alternative splice patterns and low polymorphism among class I genes. AB - The opossum major histocompatibility complex (MHC) shares a similar organization with that of non-mammals while containing a diverse set of class I genes more like that of eutherian (placental) mammals. There are 11 class I loci in the opossum MHC region, seven of which are known to be transcribed. The previously described Monodelphis domestica (Modo)-UA1 and Modo-UG display characteristics consistent with their being classical and non-classical class I genes, respectively. Here we describe the characteristics of the remaining five transcribed class I loci (Modo-UE, -UK, -UI, -UJ and -UM). All five genes have peptide-binding grooves with low or no polymorphism, contain unpaired cysteines with the potential to produce homodimer formation and display genomic organizational features that would be unusual for classical class I loci. In addition, Modo-UJ and -UM were expressed in alternatively spliced mRNA forms, including a potentially soluble isoform of Modo-UJ. Thus, the MHC region of the opossum contains a single class I gene that is clearly classical and six other class I genes each with its own unique characteristics that probably perform roles other than or in addition to antigen presentation. PMID- 19191912 TI - Virulence, immunopathology and transmissibility of selected strains of Mycobacterium tuberculosis in a murine model. AB - After encounter with Mycobacterium tuberculosis, a series of non-uniform immune responses are triggered that define the course of the infection. Eight M. tuberculosis strains were selected from a prospective population-based study of pulmonary tuberculosis patients (1995-2003) based on relevant clinical/epidemiological patterns and tested in a well-characterized BALB/c mouse model of progressive pulmonary tuberculosis. In addition, a new mouse model of transmissibility consisting of prolonged cohousing (up to 60 days) of infected and naive animals was tested. Four phenotypes were defined based on strain virulence (mouse survival, lung bacillary load and tissue damage), immunology response (cytokine expression determined by real-time polymerase chain reaction) and transmissibility (lung bacillary loads and cutaneous delayed-type hypersensitivity in naive animals).We identified four clearly defined strain phenotypes: (1) hypervirulent strain with non-protective immune response and highly transmissible; (2) virulent strain, associated with high expression of proinflammatory cytokines (tumour necrosis factor and interferon) and very low anti-inflammatory cytokine expression (interleukins 4 and 10), which induced accelerated death by immunopathology; (3) strain inducing efficient protective immunity with lower virulence, and (4) strain demonstrating strong and early macrophage activation (innate immunity) with delayed participation of acquired immunity (interferon expression). We were able to correlate virulent and transmissible phenotypes in the mouse model and markers of community transmission such as tuberculin reactivity among contacts, rapid progression to disease and cluster status. However, we were not able to find correlation with the other two phenotypes. Our new transmission model supported the hypothesis that among these strains increased virulence was linked to increased transmission. PMID- 19191913 TI - Autoantibodies to myelin basic protein (MBP) in healthy individuals and in patients with multiple sclerosis: a role in regulating cytokine responses to MBP. AB - Anti-myelin basic protein (-MBP) autoantibodies have generally been considered to be absent from sera from healthy individuals, but to be detectable in sera from some patients with multiple sclerosis (MS). However, their pathogenic role is uncertain. We demonstrate the presence of MBP-reactive autoantibodies in sera from 17 healthy individuals and 17 MS patients. The addition of MBP to the sera caused a dose-dependent deposition of MBP and co-deposition of immunoglobulin M (IgM) and fragments of complement component 3 (C3) on allogeneic monocytes. Calcium chelation abrogated the immunoglobulin deposition, indicating that formation of complement-activating immune complexes played a role in the binding process. Furthermore, MBP elicited tumour necrosis factor (TNF)-alpha and interleukin (IL)-10 production by normal mononuclear cells in the presence of serum from both patients and controls. Mononuclear cells from MS patients responded to MBP with the production of interferon (IFN)-gamma, IL-4 and IL-5, in addition to TNF-alpha and IL-10. The production of IFN-gamma and IL-5 was increased when MS serum was added rather than normal serum. Denaturation of MBP strongly inhibited MBP deposition and the MBP-induced IgM deposition and cytokine production, indicating that these events were facilitated by autoantibodies recognizing conformational epitopes on MBP. We infer that MBP-elicited TNF-alpha and IL-10 responses are promoted to equal extents by naturally occurring MBP autoantibodies and autoantibodies contained in MS sera. However, the latter seem to be more efficient in facilitating the production of IFN-gamma and IL-5. PMID- 19191914 TI - Yersinia pestis and host macrophages: immunodeficiency of mouse macrophages induced by YscW. AB - The virulence of the pathogenic Yersinia species depends on a plasmid-encoded type III secretion system (T3SS) that transfers six Yersinia outer protein (Yop) effector proteins into the cytoplasm of eukaryotic cells, leading to disruption of host defence mechanisms. It is shown in this study that Yersinia pestis YscW, a protein of the T3SS injectisome, contributes to the induction of a deficiency in phagocytosis in host macrophages and a reduction in their antigen-presenting capacity. A Y. pestis strain lacking yscW had no effect on uptake by host macrophages. In mice infected with wild-type Y. pestis, the yscW mutant or a complement strain, immunodeficiency was observed in host macrophages compared with those from uninfected mice. However, the phagocytosis and antigen presenting capacities of macrophages infected by yscW mutant strain both in vivo and in vitro were significantly higher than those by wild type strain. Consistent with this finding, when YscW was expressed in the RAW264.7 macrophage cell line, phagocytosis and antigen-presenting capacities were significantly lower than those of the control groups. These results indicate that Y. pestis YscW may directly induce immunodeficiency in murine macrophages by crippling their phagocytosis and antigen-presenting capacities. These data provide evidences to Y. pestis pathogenesis that some proteins in T3SS injectisome, such as YscW protein, might play independent roles in disrupting host defense apart from their known functions. PMID- 19191915 TI - Co-operative action of interleukin-10 and interferon-gamma to regulate dendritic cell functions. AB - Interleukin-10 (IL-10) and interferon-gamma (IFN-gamma) double producer is found in a subpopulation of T regulatory type 1 (Tr1) and T helper type 1 (Th1) cells. Consequently, it is of interest how IL-10 and IFN-gamma influence the immune system. However, few studies have addressed the co-operative action of these 'immunosuppressive' and 'immunostimulatory' cytokines. Here, we examine the effect of IL-10 combined with IFN-gamma on dendritic cell (DC) functions. Murine bone marrow-derived conventional DCs were stimulated with IL-10 and/or IFN-gamma for 24 hr. Tumour necrosis factor-alpha and IL-12 p40 production by DCs treated with both IL-10 and IFN-gamma was significantly lower than that by DCs treated with IL-10 or IFN-gamma alone. Major histocompatibility complex class II expression on DCs treated with both cytokines was attenuated compared with that on DCs treated with either cytokine alone. In contrast, levels of inducible nitric oxide synthase and indoleamine 2,3-dioxygenase, which appear to suppress T cell responses and promote tolerance, in DCs treated with both cytokines were higher than those in DCs treated with IL-10 or IFN-gamma alone. Simultaneous treatment with IL-10 and IFN-gamma significantly suppressed the ability of DCs to activate CD4+ T cells compared with treatment with either cytokine. Therefore, IL 10 and IFN-gamma co-operatively suppress the immunostimulatory functions of DCs. PMID- 19191916 TI - Major histocompatibility complex (MHC) class II but not MHC class I molecules are required for efficient control of Strongyloides venezuelensis infection in mice. AB - Strongyloides stercoralis is an intestinal nematode capable of chronic, persistent infection and hyperinfection of the host; this can lead to dissemination, mainly in immunosuppressive states, in which the infection can become severe and result in the death of the host. In this study, we investigated the immune response against Strongyloides venezuelensis infection in major histocompatibility complex (MHC) class I or class II deficient mice. We found that MHC II(-/-) animals were more susceptible to S. venezuelensis infection as a result of the presence of an elevated number of eggs in the faeces and a delay in the elimination of adult worms compared with wild-type (WT) and MHC I(-/-) mice. Histopathological analysis revealed that MHC II(-/-) mice had a mild inflammatory infiltration in the small intestine with a reduction in tissue eosinophilia. These mice also presented a significantly lower frequency of eosinophils and mononuclear cells in the blood, together with reduced T helper type 2 (Th2) cytokines in small intestine homogenates and sera compared with WT and MHC I(-/-) animals. Additionally, levels of parasite-specific immunoglobulin M (IgM), IgA, IgE, total IgG and IgG1 were also significantly reduced in the sera of MHC II(-/ ) infected mice, while a non-significant increase in the level of IgG2a was found in comparison to WT or MHC I(-/-) infected mice. Together, these data demonstrate that expression of MHC class II but not class I molecules is required to induce a predominantly Th2 response and to achieve efficient control of S. venezuelensis infection in mice. PMID- 19191919 TI - Indoor air pollution from solid biomass fuels combustion in rural agricultural area of Tibet, China. AB - In this study, we are trying to investigate the indoor air pollution and to estimate the residents' pollution exposure reduction of energy altering in rural Tibet. Daily PM(2.5) monitoring was conducted in indoor microenvironments like kitchen, living-room, bedroom, and yard in rural Tibet from December 2006 to March 2007. For kitchen air pollution, impact of two fuel types, methane and solid biomass fuels (SBFs), were compared. Questionnaire survey on the domestic energy pattern and residents' daily activity pattern was performed in Zha-nang County. Daily average PM(2.5) concentrations in kitchen, living-room, bedroom, and yard were 134.91 microg/m(3) (mean, n = 45, 95%CI 84.02, 185.80), 103.61 microg/m(3) (mean, n = 21, 95%CI 85.77, 121.45), 76.13 microg/m(3) (mean, n = 18, 95%CI 57.22, 95.04), and 78.33 microg/m(3) (mean, n = 34, 95%CI 60.00, 96.65) respectively. Using SBFs in kitchen resulted in higher indoor pollution than using methane. PM(2.5) concentrations in kitchen with dung cake, fuel wood and methane use were 117.41 microg/m(3) (mean, n = 18, 95%CI 71.03, 163.79), 271.11 microg/m(3) (mean, n = 12, 95%CI 104.74, 437.48), and 46.96 microg/m(3) (mean, n = 15, 95%CI 28.10, 65.82) respectively. Family income has significant influence on cooking energy choice, while the lack of commercial energy supply affects the energy choice for heating more. The effects of two countermeasures to improve indoor air quality were estimated in this research. One is to replace SBFs by clean energy like methane, the other is to separate the cooking place from other rooms and by applying these countermeasures, residents' exposure to particulate matters would reduce by 25-50% (methane) or 20-30% (separation) compared to the present situation. PRACTICAL IMPLICATIONS: Indoor air pollution caused by solid biomass fuels is one of the most important burdens of disease in the developing countries, which attracts the attention of environment and public health researchers, as well as policy makers. This paper gives a pilot research on the indoor air pollution and estimated the effects of some intervention policies in Tibet of China, where the living habits of the residents are quite different from other parts of the world. This work would be an important supply to the indoor air pollution studies, and would be helpful in policy making. PMID- 19191917 TI - Neither interleukin-4 receptor alpha expression on CD4+ T cells, or macrophages and neutrophils is required for protective immunity to Trichinella spiralis. AB - The T helper type 2 (Th2) mediated expulsion of the gastrointestinal nematode Trichinella spiralis requires interleukin-4 receptor alpha (IL-4Ralpha) expression on both bone-marrow-derived and non-bone-marrow-derived cells. To more definitively investigate the role of IL-4/IL-13 responsiveness in the development of protective immunity to T. spiralis, cell-specific IL-4Ralpha signalling on CD4(+) T cells (Lck(cre) IL-4Ralpha(-/flox)) and macrophages/neutrophils (LysM(cre) IL-4Ralpha(-/flox)) was analysed on the BALB/c background. Infection of wild-type and control IL-4Ralpha(-/flox) mice induced a Th2-type immune response with elevated IL-4 cytokine production, parasite-specific immunoglobulin G1 (IgG1), total IgE, intestinal mastocytosis and enteropathy. In contrast, global IL-4Ralpha-deficient BALB/c mice showed reduced worm expulsion, antibody production, intestinal mastocytosis and gut pathology. BALB/c mice generated with cell-specific deletion of IL-4Ralpha on CD4(+) T lymphocytes or macrophages/neutrophils, controlled gastrointestinal helminth infection by eliciting a protective immune response comparable to that observed with wild-type and IL-4Ralpha(-/flox) controls. Together, this shows that the development of host protective Th2 responses accompanied by parasite loss is independent of IL 4Ralpha expression on CD4(+) T cells and macrophages/neutrophils. PMID- 19191920 TI - Dampness at dorm and its associations with allergy and airways infection among college students in China: a cross-sectional study. AB - A cross-sectional study was carried out at Tianjin University campus, China, from February 21 to June10, 2006, to survey the association between dampness in dorms and allergy and airways infection among college students. The health and dampness condition were self-reported by 3436 students living in 1511 dorm rooms located in 13 buildings on the campus. The buildings were selected according to their positions, construction periods and occupant densities. The allergy and airways infection symptoms involved wheezing, dry cough during night, rhinitis, eczema, cold/flu, ear inflammation, pneumonia and tuberculosis. The indoor moisture signs were mould/damp spots on walls, ceilings and floors; suspected or ever happened water damage; condensation on windowpane in winter and odours perceived by subjects themselves. This study showed there was significantly positive association between condensation and dry cough. Eczema was often reported in rooms with suspected moisture problem. Dampness was a significantly risk factor for common cold. PRACTICAL IMPLICATIONS: This paper indicated that dampness problem at dorms of Chinese students was a risk factor in irritating allergic symptoms, and hence there is a need for dorm environment improvement. The ventilation and microbiology problems in dorm environment corresponding to dampness should be further studied, especially when it is associated to occupants' health. PMID- 19191918 TI - CD28 and inducible costimulator (ICOS) signalling can sustain CD154 expression on activated T cells. AB - The biological outcome of receptor-mediated signalling often depends on the duration of engagement. Because CD40 signalling is controlled by the regulated expression of its ligand, CD154, the mechanisms that regulate CD154 expression probably determine the strength and duration of CD40 signalling. Here, we demonstrate that CD154 expression on the surface of mouse CD4 T cells can be separated into an early phase, occurring between 0 and 24 hr after T-cell activation, and a later extended phase, occurring after 24 hr. The early phase of CD154 expression did not require costimulation and was probably influenced by the strength of T-cell receptor (TCR) signalling alone. However, later CD154 expression was highly dependent on costimulation through either CD28 or inducible costimulator (ICOS). Although CD28 signalling interleukin (IL)-2 secretion, ICOS not, suggesting that costimulation enhance CD154 expression independently of IL-2 production. In fact, anti-CD28 treatment could still induce late-phase CD154 on anti-CD3-stimulated CD4 T cells expressing a mutated form of CD28 that not lead to the induction of IL-2. However, this CD154 induction was somewhat weaker than that of wild-type CD28-expressing cells, suggesting that direct signalling and IL 2-mediated signalling co-operatively responsible for the levels of CD154 induced by CD28. Finally, we show that the second phase of CD154 expression negatively regulated B-cell terminal differentiation and antibody secretion. These results demonstrate that TCR signalling and costimulation each regulate different phases of CD154 expression and control the biological outcome of CD40 signalling on B cells. PMID- 19191922 TI - Citations, impact factors, and Indoor Air: a look behind the numbers. PMID- 19191921 TI - Molds and mycotoxins in dust from water-damaged homes in New Orleans after hurricane Katrina. AB - Dust collected in New Orleans homes mold-contaminated because of the flooding after hurricane Katrina was analyzed for molds and mycotoxins. The mycoflora was studied by cultivation and quantitative PCR for selected molds. The most commonly found mold taxa were Aspergillus, Cladosporium, and Penicillium. Verrucarol, a hydrolysis product of macrocyclic trichothecenes predominately produced by Stachybotrys spp. was identified in three dust samples by gas chromatography tandem mass spectrometry, and sterigmatocystin (produced by various Aspergillus spp.) was found in two samples by high pressure liquid chromatography-tandem mass spectrometry. This is the first demonstration of mycotoxins in Katrina-associated dust samples. The analytical methods used represent valuable tools in further studies on bioaerosol exposure and health risks. PRACTICAL IMPLICATIONS: In the aftermath of natural disasters like hurricane Katrina water-damages on infrastructure and public and private property are often associated with health risks for remediation workers and returning residents. In the case of New Orleans evaluations of health hazards, health studies, and assessments of bioaerosol have been conducted previously. However, until now mycotoxins have not been addressed. Our study shows, for the first time, the presence of mycotoxins in dust collected in houses in New Orleans mold-contaminated because of the hurricane Katrina. The results may highlight the potential health threats posed by mold aerosols in post disaster inhabited areas. PMID- 19191923 TI - A one-dimensional analytical model for airborne contaminant transport in airliner cabins. AB - Quick information on airborne infectious disease transmission in airliner cabins is essential to reduce the risk of infection of passengers and crew members. This investigation proposed a one-dimensional analytical model that can predict the longitudinal transmission of airborne contaminants or disease viruses inside an airliner cabin. The model considered both diffusive and convective transport of contaminants in the longitudinal direction of the cabin but assumed complete mixing of contaminants in the cabin cross-section. The effect of recirculation of the cabin air and efficiency of the high-efficiency particulate air (HEPA) filters is also considered in the model. The analytical solution for the one dimensional contaminant transport model is obtained by using the principle of superposition and the method of separation of variables. The analytical solutions agree well with the computational fluid dynamics (CFD) results. The coupling of a CFD model with the one-dimensional analytical model could capture the impact of local airflow on contaminant transport. This analytical model has been used for analyzing contaminant transport in a 30-row all-economy-class airliner cabin with minimal computing effort. PRACTICAL IMPLICATIONS: The paper presents a new one dimensional analytical model that can provide quick information on global airborne contaminant transmissions in airliner cabins for effective response plans. The model can be used to study the effects of air exchange rates, recirculation, efficiency of the high-efficiency particulate air (HEPA) filters and longitudinal airflow on airborne contaminant transport in airliner cabins with minimal computing effort. PMID- 19191924 TI - Magnitude of indoor NO2 from biomass fuels in rural settings of Ethiopia. AB - Half of the world's population and about 80% of households in Sub-Saharan Africa depend on biomass fuels. Indoor air pollution due to biomass fuel combustion may constitute a major public health threat affecting children and women. The purpose of this study was to measure levels of indoor NO(2) concentration in homes with under-five children in rural Ethiopia. The study was undertaken in the Butajira area in Ethiopia from March 2000 to April 2002. 24-h samples were taken regularly at about three month intervals in approximately 3300 homes. Indoor air sampling was done using a modified Willems badge. For each sample taken, an interview with the mother of the child was performed. A Saltzman colorimetric method using a spectrometer calibrated at 540 nm was employed to analyze the mass of NO(2) in field samples. Wood, crop residues and animal dung were the main household fuels. The mean (s.d.) 24-h concentration of NO(2) was 97 microg/m(3) (91.4). This is more than double the currently proposed annual mean of WHO air quality guideline. Highland households had significantly higher indoor NO(2) concentration. This study demonstrates high levels of indoor NO(2) in rural homes of Ethiopia. PRACTICAL IMPLICATIONS: Respiratory infection is a major cause of morbidity and mortality, globally. Acute respiratory symptoms are also related to high levels of air pollution. Interventions aimed at reducing exposure to indoor air pollution should focus on cooking and heating practices in developing countries. This study is not undermining the role of other biomass smoke constituents in determining respiratory infections. PMID- 19191925 TI - Improving indoor air quality for poor families: a controlled experiment in Bangladesh. AB - The World Health Organization's 2004 Global and Regional Burden of Disease Report estimates that acute respiratory infections from indoor air pollution (pollution from burning wood, animal dung, and other bio-fuels) kill a million children annually in developing countries, inflicting a particularly heavy toll on poor families in South Asia and Africa. This paper reports on an experiment that studied the use of different fuels in conjunction with different combinations of construction materials, space configurations, cooking locations, and household ventilation practices (use of doors and windows) as potentially-important determinants of indoor air pollution. Results from controlled experiments in Bangladesh were analyzed to test whether changes in these determinants can have significant effects on indoor air pollution. Analysis of the data shows, for example, that pollution from the cooking area is transported into living spaces rapidly and completely. Furthermore, it is important to factor in the interaction between outdoor and indoor air pollution. Hence, the optimal cooking location should take 'seasonality' in account. Among fuels, seasonal conditions seem to affect the relative severity of pollution from wood, dung, and other biomass fuels. However, there is no ambiguity about their collective impact. All are far dirtier than clean (LPG and Kerosene) fuels. The analysis concludes that if cooking with clean fuels is not possible, then building the kitchen with permeable construction material and providing proper ventilation in cooking areas will yield a better indoor health environment. PRACTICAL IMPLICATIONS: Several village-level measures could significantly reduce IAP exposure in Bangladesh. All would require arrangements and the assert of male heads-of-household: negotiated bulk purchases of higher cost, cleaner fuels; purchase of more fuel-efficient stoves; peripheral location of cooking facilities; building the kitchen with permeable construction material; rotation of women in cooking roles, to reduce their exposure; and ventilation of smoke through a stack tall enough to disperse smoke over a relatively broad area. It is expected that village men and women will agree to these measures if they become convinced that IAP poses a serious risk to health, and their actions will significantly reduce the risk. The keys to success are effective public education about the sources and risks of IAP, and financial and technical assistance for changes in cooking arrangements. PMID- 19191926 TI - Real-time or faster-than-real-time simulation of airflow in buildings. AB - Real-time flow simulation is crucial for emergency management in buildings, such as fire and accidental or intentional release of chemical/biological agents (contaminants). The simulation results can then be used to impose proper measures to minimize casualties. Computational fluid dynamics (CFD) is accurate, but too time-consuming. Nodal models are fast, but not informative. To obtain a quick and informative solution, this study proposes an intermediate approach between nodal models and CFD by introducing a fast fluid dynamics (FFD) method. This investigation used the FFD methods with and without turbulence treatments to study systematically four basic flows in buildings, and compared the numerical results with the corresponding CFD results and the data from the literature. The results show that, on one hand, the FFD can offer much richer flow information than nodal models, but less accurate results than CFD. On the other hand, the FFD is 50 times faster than the CFD. The results also show that the FFD with the laminar assumption has the best overall performance as regards both accuracy and speed. It is possible to conduct faster-than-real-time flow simulations with detailed flow information by using the FFD method. PRACTICAL IMPLICATIONS: The paper introduces a fast fluid dynamics (FFD) method, which can simulate airflow and contaminant dispersion in buildings with real-time or faster-than-real-time speed and provide informative solutions. As an intermediate approach between nodal models and the computational fluid dynamics (CFD), the FFD can be a very useful tool for emergency management in case of fire and accidental or intentional release of chemical or biological agents in a building or around the buildings. The FFD can also be used as a preliminary test tool for quick assessment of indoor airflows before a detailed CFD analysis. PMID- 19191927 TI - Health evaluation of volatile organic compound (VOC) emission from exotic wood products. AB - The purpose of this study was to measure and evaluate the impact of the emissions of selected products of exotic wood on health. Ten products were screened for chemical compounds, and five of the most used products which emitted more than 800 microg/kg were selected for further quantitative analyses by climate chamber measurement (iroko, ramin, sheesham, merbau, and rubber tree). Samples of exotic wood (rubber tree and belalu) were further analyzed for emission of chemical compounds by migration into artificial saliva and for content of pesticides and allergenic natural rubber latex (NR latex) (rubber tree). The toxicological effects of all substances identified were evaluated and the lowest concentrations of interest (LCI) assessed. An R-value was calculated for each wood product (R value below 1 is considered to be unproblematic as regards health). Emission from the evaluated exotic wood only took place to a very limited extent. None of the selected products, under the chosen rating system, is likely to cause adverse health effects. Products with surface treatment might pose a problem if used as kitchen utensils, as children's toys, or when they are in close contact with the skin for a long time. PRACTICAL IMPLICATIONS: The authors investigated the chemical emissions from selected products from exotic wood by climate chamber measurement. Quantitative chemical analyses of emissions from the five most used exotic products in Denmark were performed, and all chemical compounds found were evaluated toxicologically. Emission from the evaluated exotic wood was very limited. None of the products is likely, under our exposure conditions, to cause health problems in relation to indoor air. PMID- 19191928 TI - Sarcoidosis, asthma, and asthma-like symptoms among occupants of a historically water-damaged office building. AB - Sarcoidosis is a granulomatous disease of unknown etiology with evidence of association with exposure to microbial agents. In June 2006, we investigated a sarcoidosis cluster among office workers in a water-damaged building. In the course of the investigation, we became aware of a high rate of respiratory complaints including asthma and asthma-like symptoms. We conducted case finding for physician-diagnosed sarcoidosis and asthma and administered a health questionnaire survey and pulmonary function tests (PFTs) to consenting occupants. We compared prevalence ratios (PRs) to the Environmental Protection Agency's Building Assessment Survey and Evaluation study (BASE) and the National Health and Nutrition Examination Survey (NHANES). We identified six sarcoidosis cases. The current building prevalence is 2206 cases/100,000 population, elevated, compared with the US population range of <1-40 cases/100,000. Of current occupants, 77% (105) participated in the health questionnaire survey and 64% (87) in PFTs. Physician-diagnosed asthma was elevated, compared with the US adult population. Adult asthma incidence was 3.3/1000 person-years during the period before building occupancy and 11.5/1000 person-years during the period after building occupancy. Comparisons with US office workers (BASE) yielded elevated PRs for shortness of breath [PR, 9.6; 95% confidence interval (CI), 6.1-15.2], wheeze (PR, 9.1; 95% CI 5.6-14.6), and chest tightness (PR, 5.1; 95% CI 2.8-9.0). PFT results supported reports of respiratory symptoms and diagnoses. Based on our findings building occupants were relocated. PRACTICAL IMPLICATIONS: The remission of occupational asthma caused by certain known antigens improves with early diagnosis and removal from exposure. As a suspected antigen-mediated disease, sarcoidosis might also benefit if affected persons are isolated from continued exposure. Our investigation identified a high prevalence of new-onset sarcoidosis, and asthma among workers of a water damaged building with a history of indoor environmental quality complaints. Removal of all individuals from such environments until completion of building diagnostics, environmental sampling and complete remediation is a prudent measure when feasible. PMID- 19191929 TI - A TRP channel is expressed in Spodoptera littoralis antennae and is potentially involved in insect olfactory transduction. AB - The molecular characterization of post-receptor actors involved in insect olfactory transduction has yet to be understood. We have investigated the presence of a Transient Receptor Potential (TRP) channel in the peripheral olfactory system of the moth Spodoptera littoralis. A cDNA encoding a Lepidopteran TRP channel (TRPgamma) was identified by analysis of a male-antennal EST database and subsequently cloned by RACE PCR. In adult males, the TRPgamma transcript was detected in antennae, at the base of olfactory sensilla. Moreover, TRPgamma was observed in antennae in both pupal and adult stages. This work is the first step in understanding the involvement of TRPgamma in signalling pathways involved in the development and function of the insect olfactory system. PMID- 19191930 TI - Post-chemotherapy nerve-sparing retroperitoneal lymph node dissection for advanced germ cell tumor. AB - OBJECTIVE: To report our experience with post-chemotherapy nerve-sparing retroperitoneal lymph node dissection (RPLND) for advanced germ cell tumor (GCT). METHODS: Between 1994 and 2008, 92 patients with advanced GCT underwent RPLND after multiple treatments with systemic chemotherapy at our institution. A nerve sparing RPLND was carried out in 78 patients (84.8%; median age 32 years). Of them, 19 had a seminoma and 59 had a non-seminoma. RESULTS: Lumbar splanchnic nerves controlling ejaculatory function were macroscopically preserved during RPLND. Bilateral and unilateral lumbar splanchnic nerves were preserved in 40 patients and 38 patients, respectively. Sixty-five patients could be evaluated for ejaculation. Fifty-four patients (83.1%) achieved antegrade ejaculation with a median postoperative interval of 3 months (range: 1-10 months). Twenty-eight patients (28/30: 93.3%) and 26 patients (26/35: 74.3%) undergoing bilateral and unilateral nerve-sparing RPLND had antegrade ejaculation, respectively (P = 0.041). Only two patients (2.6%) had mediastinal and retroperitoneal recurrences during a median follow-up of 42 months (range: 1-138 months), respectively. However, these patients were cured by chemotherapy and surgery. CONCLUSIONS: Post chemotherapy nerve-sparing RPLND preserves ejaculatory function in the majority of patients with advanced GCT without increasing the risk of local recurrence. PMID- 19191931 TI - Relying on machines to do the breathing. PMID- 19191932 TI - Incarceration and the tasks of grief: a narrative review. AB - AIM: This study is a report of a narrative review to explore the challenges facing prisoners and the corrections system in the presence of the death of a significant person to the prisoner. BACKGROUND: Death of a loved one is an important challenge, amplified for incarcerated men. There are unique aspects of incarceration that prevent prisoners from having access to usual ritual expressions and support structures. DATA SOURCES: A search of the CINAHL, ProQuest Medical, PubMed, EBSCO and COCHRANE databases was conducted for papers published from 1998-2007. The search terms were bereavement and prison nursing. A hand search of material specific to grief and incarceration was also undertaken. REVIEW METHODS: A narrative technique involving reading, writing, thinking, interpreting, arguing and justifying was used to synthesize the material and create a convincing and cohesive story. RESULTS: Limited research is available specifically addressing the grief experience of incarcerated individuals or the impact of unresolved grief on recidivism. However, a number of potential challenges to the grieving process in the prison system are identified in the literature, such as the prison culture of toughness and limited options for funeral attendance. CONCLUSION: Whilst the literature is scant, it is clear that issues of masculinity and culture have a strong impact on the ability of incarcerated men to resolve grief issues. More research is required to understand the impact of this on issues, such as recidivism. In the meantime, prison nurses have an important role to play in supporting prisoners who have lost a loved one during their incarceration. PMID- 19191934 TI - The use of technology in cancer care: applying Foucault's ideas to explore the changing dynamics of power in health care. AB - AIM: This study is a report to identify the utility of a hand-held side-effect monitoring system for people receiving chemotherapy in the home care setting. BACKGROUND: Increasingly, health care is being provided in people's own homes and communities rather than in hospitals. This has driven the development of technologies which support patients in the home environment. The meaning of such technologies can be explored from a Foucauldian perspective to shed light on how they enable new forms of medical surveillance. METHOD: An intervention study was performed in 2006 using new technologies for people receiving chemotherapy. Questionnaires were completed by 56 people affected by cancer who used the new technology; 12 of these people were then interviewed. Secondary analysis of the interview data is presented in this paper, drawing on Foucault's writing about surveillance and power in medical settings. FINDINGS: The interview transcripts contain numerous examples of people affected by cancer reflecting on issues such as power and surveillance in cancer care. While these terms are ordinarily considered to reflect negative elements of care, they were used by participants in an empowering manner. CONCLUSION: Theoretical insights can help nurses to think critically about the advances of technology. In particular, there are implications for how nurses consider the relationship of technology to patients and for power dynamics in healthcare relationships. We suggest that there is a need to problematize and celebrate the growth of technologically-driven health surveillance. PMID- 19191935 TI - Staff-student relationships and their impact on nursing students' belongingness and learning. AB - AIM: This paper is a report of selected findings from a study exploring the relationship between belongingness and placement experiences of preregistration nursing students. BACKGROUND: Staff-student relationships are an important influence on students' experiences of belongingness and their clinical learning. The need to belong is universal and pervasive, exerting a powerful influence on thought processes, emotions, behaviour, health and happiness. People deprived of belongingness are more likely to experience diminished self-esteem, increased stress and anxiety, depression and a decrease in general well-being. Nursing students' motivation and capacity to learn, self-concept, confidence, the extent to which they are willing to question or conform to poor practice and their future career decisions are influenced by the extent to which they experience belongingness. METHOD: During 2006, 18 third year students from two Australian universities and one United Kingdom university participated in in-depth semi structured interviews. Data were analysed thematically. FINDINGS: Participants described placement experiences spanning a continuum from those promoting a high degree of belongingness to those provoking intense feelings of alienation. Staff student relationships (including receptiveness, inclusion/exclusion, legitimization of the student role, recognition and appreciation, challenge and support) were the most important influence on students' sense of belonging and learning. Similarities between sites were remarkable, despite the differences in healthcare and higher education systems. CONCLUSION: Staff-student relationships are key to students' experience of belongingness. Understanding the types of interactions and behaviours that facilitate or impede students' belongingness and learning are essential to the creation of positive clinical experiences. PMID- 19191936 TI - Innovative behaviour: the effect of structural and psychological empowerment on nurses. AB - AIM: This paper is a report of a study to investigate the relationship between structural empowerment/psychological empowerment and innovative behaviour. BACKGROUND: Innovative behaviour is essential for nurses in today's dynamic healthcare systems. Empowerment has become an increasingly important factor in predicting innovative behaviour. Structural empowerment refers to power based on the employee's position in the organization, while psychological empowerment consists of the fundamental personal convictions that employees have about their role in the organization. METHODS: A cross-sectional correlational survey was conducted in The Netherlands in 2007 with 519 Registered Nurses. The instruments used were the Conditions of Work Effectiveness Questionnaire II, the Psychological Empowerment Instrument and the Questionnaire Innovative Behaviour. Four hypotheses were tested using descriptive statistics, bivariate and multiple regression and one-way analysis of variance. RESULTS: Structural and psychological empowerment were statistically significant predictors of innovative behaviour. Informal power and impact were the most relevant determinants of innovative behaviour, the latter the strongest. Psychological empowerment functioned as a mediator between structural empowerment and innovative behaviour. The expected moderating effect of structural empowerment was not proven. CONCLUSION: Organizations need to create the right conditions to be able to strengthen nurses' empowerment. Networking is an important skill which should be incorporated in the nursing education. Future research should take into consideration nursing culture and personality traits as determining factors. Replication of this study in another setting and a study examining causality are recommended so that additional empirical evidence may be obtained. PMID- 19191938 TI - The impact of neuropathic pain on relationships. AB - AIM: This paper is a report of a study exploring the impact of neuropathic pain on family, social and working relationships among patients at a pain clinic serving a large urban area. BACKGROUND: Neuropathic pain is a particularly distressing type of chronic pain which is extremely difficult to manage successfully. It produces a range of unpleasant symptoms and adversely affects patients' quality of life, but little is known about its personal impact. METHOD: A descriptive and exploratory approach was used and 10 participants participated in three focus groups in 2005. Because of the low response rate of 20% from the initial sample, a second sample of 16 patients was invited to participate. However, only one person responded and therefore it was not possible to convene an additional group. FINDINGS: The unpleasant and bizarre nature of neuropathic pain underpinned much of its impact in terms of respondents' difficulties in maintaining a range of relationships. For closer relationships, key difficulties centred on the reduction in quality and/or number of personal relationships. For more distant relationships and those with professionals, frustration at the invisibility of their pain and their own failure to communicate symptoms and its consequences were central. CONCLUSION: More extensive work is needed to improve our understanding of how neuropathic pain is experienced, how it affects close and more distant kinds of relationships, and how healthcare professionals might best support people with persistent neuropathic pain to maintain personal and social relationships, and to communicate their pain effectively. PMID- 19191937 TI - Nurse practitioners substituting for general practitioners: randomized controlled trial. AB - AIM: This paper is a report of a study conducted to evaluate process and outcomes of care provided to patients with common complaints by general practitioners or specially trained nurse practitioners as first point of contact. BACKGROUND: Studies in the United States of America and Great Britain show that substituting nurse practitioners for general practitioners results in higher patient satisfaction and higher quality of care. As the American and British healthcare system and settings differ from that in The Netherlands, a Dutch trial was conducted. METHODS: A total of 1501 patients in 15 general practices were randomized to consultation by a general practitioner or a nurse practitioner. Data were collected over a 6-month period in 2006 by means of questionnaires, extracting medical records from practice computer systems and recording the length of consultations. FINDINGS: In both groups, the patients highly appreciated the quality of care. No statistically significant differences were found in health status, medical resource consumption and compliance of practical guidelines in primary care in The Netherlands. Patients in the NP intervention group were more often invited to re-attend, had more follow-up consultations and their consultations took statistically significantly longer. CONCLUSION: Nurse practitioners and general practitioners provide comparable care. Our findings support an increased involvement of specially trained nurse practitioners in the Dutch primary care and contribute to knowledge of the effectiveness of care provision by nurse practitioners from a national and international perspective. PMID- 19191939 TI - Jordanian nurses' knowledge and practice of breast self-examination. AB - AIM: This paper is a report of a study to evaluate the knowledge and practice of Jordanian nurses in relation to breast self-examination. BACKGROUND: Studies have shown that women who have learned about breast self-examination have positive attitudes toward breast cancer and practise breast self-examination more frequently, and that nurses who teach their clients about methods of early detection and breast self-examination are more knowledgeable about breast cancer screening and breast self-examination techniques than those who do not. Therefore, it is important to understand nurses' knowledge about breast cancer and its early detection. METHODS: A cross-sectional design was used, with a convenience sample of 347 Registered Nurses at three large cities in Jordan (response rate 95%). Data collection took place in 2005 using a self administrated questionnaire with three parts and based on the American Cancer Society's guidelines: demographics, knowledge, and practice of breast self examination. RESULTS: Nurses reported high levels of knowledge of breast self examination (M = 7.6, SD. 2.7). A high proportion of nurses reported doing breast self-examination in the past 12 months (85%), but only 17.7% reported doing so on a monthly basis. None of the demographic characteristics was found to be associated with the practice of breast self-examination. CONCLUSION: More health education about monthly breast self-examination and prevention strategies is needed for nurses and their women patients, especially for Arabic women. PMID- 19191940 TI - The effect of three positioning methods on patient outcomes after cardiac catheterization. AB - AIM: This paper is a report of a study to investigate the effect of three positioning protocols on back pain, heart rate, blood pressure and vascular complications after cardiac catheterization. BACKGROUND: After cardiac catheterization, bed rest is prescribed in order to minimize vascular complications, but this often leads to back pain and other complications, such as hemodynamic instability. METHODS: A three-group quasi-experimental design was used in this study, which was conducted in 2006. A convenience sample of 105 patients was randomly assigned to either the control or the two experimental groups (A and B). The control group received routine care. Group B was treated only with modified positioning and group A with modified positioning and a pillow under their body. Back pain, heart rate, arterial blood pressure, haematoma formation and bleeding were measured at regular time intervals. FINDINGS: The control group experienced higher levels of pain after 3, 6, 8 hours and the morning after catheterization. The level of pain in group B was also higher than in group A at 3 hours after the procedure. Mean heart rate and blood pressure were lower in the experimental groups compared with the control group at 6 and 8 hours after catheterization. No statistically significant difference between the three groups regarding the amounts of overall bleeding and overall haematoma formation was observed. CONCLUSION: Changing position in bed and using a supportive pillow during the early hours after cardiac catheterization can effectively minimize pain and hemodynamic instability without increasing vascular complications. PMID- 19191941 TI - Clients' experiences of living at home with a mechanical ventilator. AB - AIM: This paper reports on a study of how clients experience living with home mechanical ventilation and how they experience care and supervision of healthcare personnel. BACKGROUND: The number of people living at home with mechanical ventilators is increasing, and this is considered a successful approach to reducing incapacity and mortality. METHOD: Qualitative interviews were conducted with 10 service users in 2006. The informants were 18-75 years old and had varying diagnoses and levels of functioning. The interviews were tape recorded, transcribed and analysed by qualitative content analysis. FINDINGS: Two main themes emerged: Theme 1. Having a home ventilator enhances quality of life--a life worth living. The ventilator treatment builds up strength and improves well being. Participants emphasized that it was important to feel in control of their own situation and had an overriding wish to live a normal and active life; Theme 2. Competence and continuity of healthcare personnel are factors for success. The experience was that competence and follow-up by healthcare personnel varied, and that good quality teaching and information were important. CONCLUSION: Users of home mechanical ventilators should be active partners in their own care so that their experience is taken into account. It is important for clients having home mechanical ventilation to be empowered and have control in their daily lives, as well as having competent caregivers and continuity of care. PMID- 19191942 TI - Care dependency of hospitalized children: testing the Care Dependency Scale for Paediatrics in a cross-cultural comparison. AB - AIM: This paper is a report of a study to examine the psychometric properties of the Care Dependency Scale for Paediatrics in Germany and Egypt and to compare the care dependency of school-age children in both countries. BACKGROUND: Cross cultural differences in care dependency of older adults have been documented in the literature, but little is known about the differences and similarities with regard to children's care dependency in different cultures. METHOD: A convenience sample of 258 school-aged children from Germany and Egypt participated in the study in 2005. The reliability of the Care Dependency Scale for Paediatrics was assessed in terms of internal consistency and interrater reliability. Factor analysis (principal component analysis) was employed to verify the construct validity. A Visual Analogue Scale was used to investigate the criterion-related validity. FINDINGS: Good internal consistency was detected both for the Arabic and German versions. Factor analysis revealed one factor for both versions. A Pearson's correlation between the Care Dependency Scale for Paediatrics and Visual Analogue Scale was statistically significant for both versions indicating criterion-related validity. Statistically significant differences between the participants were detected regarding the mean sum score on the Care Dependency Scale for Paediatrics. CONCLUSION: The Care Dependency Scale for Paediatrics is a reliable and valid tool for assessing the care dependency of children and is recommended for assessing the care dependency of children from different ethnic origins. Differences in care dependency between German and Egyptian children were detected, which might be due to cultural differences. PMID- 19191943 TI - Using an algorithmic model to reveal individually variable movement decisions in a wintering sea duck. AB - 1. Many migratory birds are assumed to remain fairly stationary during winter. However, recent research indicates that mid-winter movements are evident in a variety of bird species, and the factors causing individuals to move are poorly understood. 2. We examined the winter movements of 95 individual king eiders (Somateria spectabilis, L.) tracked with satellite transmitters in the Bering Sea between 2002 and 2006 to explore whether environmental factors such as day length, location, sea ice, and habitat quality could explain the occurrence of winter movements longer than 50 km. 3. We used a novel algorithmic random forest model to assess the importance of variables predicting whether a bird remained or departed from a wintering site. 4. We found extremely high individual variability in winter movement decisions by king eiders, and the individual bird was the most important variable followed by location, date, and sea ice concentration. 5. We conclude that individual strategies exist that interact with environmental conditions to form multiple movement patterns. 6. While a minor proportion of winter movements may be forced by environmental conditions, we propose that many winter movements may be of an exploratory nature where individuals aim to acquire information about alternative wintering sites that may enhance their survival probability at some point in time when environmental fluctuation renders their preferred wintering site unsuitable. PMID- 19191944 TI - A nested real-time PCR assay has an increased sensitivity suitable for detection of viruses in aerosol studies. AB - AIMS: Influenza is commonly spread by infectious aerosols; however, detection of viruses in aerosols is not sensitive enough to confirm the characteristics of virus aerosols. The aim of this study was to develop an assay for respiratory viruses sufficiently sensitive to be used in epidemiological studies. METHOD: A two-step, nested real-time PCR assay was developed for MS2 bacteriophage, and for influenza A and B, parainfluenza 1 and human respiratory syncytial virus. Outer primer pairs were designed to nest each existing real-time PCR assay. The sensitivities of the nested real-time PCR assays were compared to those of existing real-time PCR assays. Both assays were applied in an aerosol study to compare their detection limits in air samples. CONCLUSIONS: The nested real-time PCR assays were found to be several logs more sensitive than the real-time PCR assays, with lower levels of virus detected at lower Ct values. The nested real time PCR assay successfully detected MS2 in air samples, whereas the real-time assay did not. SIGNIFICANCE AND IMPACT OF THE STUDY: The sensitive assays for respiratory viruses will permit further research using air samples from naturally generated virus aerosols. This will inform current knowledge regarding the risks associated with the spread of viruses through aerosol transmission. PMID- 19191945 TI - Genetic analysis of Saccharomyces cerevisiae strains isolated from palm wine in eastern Nigeria. Comparison with other African strains. AB - AIMS: To study the yeast diversity of Nigerian palm wines by comparison with other African strains. METHODS AND RESULTS: Twenty-three Saccharomyces cerevisiae strains were obtained from palm wine samples collected at four locations in eastern Nigeria, and characterized using different molecular techniques: internal transcribed spacer restriction fragment length polymorphism and sequence analysis, pulsed field gel electrophoresis, inter delta typing and microsatellite multilocus analysis. These techniques revealed that palm wine yeasts represent a group of closely related strains that includes other West African isolates (CBS400, NCYC110, DVPG6044). Population analysis revealed an excess of homozygote strains and an allelic richness similar to wine suggestive of local domestication. Several other African yeast strains were not connected to this group. Ghana sorghum beer strains and other African strains (DBVPG1853 and MUCL28071) displayed strikingly high relatedness with European bread, beer or wine strains, and the genome of strain MUCL30909 contained African and wine-type alleles, indicating its hybrid origin. CONCLUSIONS: Nigerian palm wine yeast represents a local specific yeast flora, whereas a European origin or hybrid was suspected for several other Africa isolates. SIGNIFICANCE AND IMPACT OF THE STUDY: This study presents the first genetic characterization of an autochthonous African palm wine yeast population and confirms the idea that human intervention has favoured yeast migration. PMID- 19191946 TI - Characterization of lactosporin, a novel antimicrobial protein produced by Bacillus coagulans ATCC 7050. AB - AIMS: To characterize the antimicrobial protein produced by Bacillus coagulans used in the probiotic dietary supplement (Lactospore) Probiotic, Sabinsa Corp., Piscataway, NJ, USA). METHODS AND RESULTS: Bacillus coagulans ATCC 7050 was grown at 37 degrees C for 18 h. The cell free supernatant was concentrated 10-fold (lactosporin preparation, LP). The antimicrobial activity of LP was confirmed against Micrococcus luteus ATCC 10420 in a well diffusion assay. The proteinaceous nature of LP was determined following exposure to different enzymes. The activity of LP was pH-dependent but stable to heat. The isoelectric point of LP was determined to be 3.5-4.0. PCR analyses showed no similarity between lactosporin and known antimicrobial proteins produced by the Bacillus spp. CONCLUSIONS: Lactosporin is a novel antimicrobial protein. Initial characterization indicates that it may fall outside of the conventional classification of class I and II bacteriocins. Loss of activity after exposure to a number of proteolytic enzymes and lipase suggest that lactosporin may posses a lipid moiety which contributes to its inhibitory activity. SIGNIFICANCE AND IMPACT OF THE STUDY: The unique characteristics of lactosporin, including its antimicrobial activity against pathogenic micro-organisms, indicate that it may have potential for application in foods and personal care products. PMID- 19191947 TI - PCR screening of 3-amino-5-hydroxybenzoic acid synthase gene leads to identification of ansamycins and AHBA-related antibiotic producers in Actinomycetes. AB - AIMS: The 3-amino-5-hydroxybenzoic acid (AHBA) synthase is one of the essential and unique enzymes for AHBA biosynthesis. The possibility of screening for ansamycin or AHBA-related antibiotic-producing strains from Actinomycetes by targeting an AHBA synthase gene was explored. METHODS AND RESULTS: A pair of degenerated primers designed according to the conserved regions of five known AHBA synthases was used to detect AHBA synthase genes within the genomic DNA of Actinomycetes. PCR screening resulted in obtaining 33 AHBA synthase gene-positive strains from 2000 newly isolated Actinomycetes. Phylogenetic analysis of these gene fragments along with those involved in the biosynthesis of structurally determined ansamycins showed that the genes with close phylogenetic relationships might be involved in the biosynthesis of compounds with the same/similar structures. Four strains have been proved to be actual geldanamycin or rifamycin producers by chemical characterization of their fermentation products. CONCLUSIONS: The results confirmed the feasibility of using the AHBA synthase gene as a probe in polymerase chain reaction (PCR) screening of ansamycin or AHBA related antibiotic-producing strains. SIGNIFICANCE AND IMPACT OF THE STUDY: The PCR screening of AHBA synthase gene represents a direct and sensitive molecular method for rapid detection of AHBA-related antibiotic-producing strains. PMID- 19191948 TI - Cyanophycin-degrading bacteria in digestive tracts of mammals, birds and fish and consequences for possible applications of cyanophycin and its dipeptides in nutrition and therapy. AB - AIMS: To determine the susceptibility of cyanophycin granule polypeptide (CGP) to degradation by several mammalian, avian and fish gut flora. METHODS AND RESULTS: Samples of gut flora were investigated for the occurrence of bacteria capable of CGP degradation. With all samples, a complete anaerobic degradation of CGP was achieved over incubation periods of only 12-48 h at 37 degrees C. CGP-degrading bacteria were detected in all samples, and they occurred in particular high titres in caecum flora from rabbit and sheep and in the digestive tract of carp fish. A total of 62 axenic cultures were isolated. All degraded CGP aerobically, 46 of them degraded CGP also anaerobically over incubation periods ranging from 24 h to 7 days. HPLC analysis revealed that all isolates degraded CGP to its constituting dipeptides. Eight strains were identified by 16S rRNA gene sequencing and were affiliated to the genera Bacillus, Brevibacillus, Pseudomonas, Streptomyces and Micromonospora. CONCLUSIONS: These data demonstrate for the first time the occurrence of a natural niche for CGP in the digestive tracts of animals. SIGNIFICANCE AND IMPACT OF THE STUDY: The biodegradability of CGP by gut flora provides a first confirmation for the potential applications of CGP and its dipeptides in nutrition and therapy as highly bio-available sources for arginine, lysine, aspartate and possibly also other amino acids. PMID- 19191949 TI - Effect of the fermentation pH on the storage stability of Lactobacillus rhamnosus preparations and suitability of in vitro analyses of cell physiological functions to predict it. AB - AIMS: To investigate how cell physiological functions can predict the stability of freeze-dried probiotics. In addition, the effect of the fermentation pH on the stability of probiotics was investigated. METHODS AND RESULTS: Fermenter-grown (pH 5.8 or 5.0) Lactobacillus rhamnosus cells were freeze-dried and their survival was evaluated during storage at 37 degrees C, in apple juice and during acid [hydrochloric acid (HCl) and malic acid] and bile exposure. Cells grown at pH 5.0 were generally coping better with acid-stress than cells grown at pH 5.8. Cells were more sensitive to malic acid compared with HCl. Short-term stability results of Lact. rhamnosus cells in malic acid correlated well with the long-term stability results in apple juice, whereas the results of cell membrane integrity studies were in accordance with bile exposure results. CONCLUSIONS: Malic acid exposure can prove useful in evaluating the long-term stability of probiotic preparations in apple juice. Fermentation at reduced pH may ensure a better performance of Lact. rhamnosus cells during the subsequent acid-stress. SIGNIFICANCE AND IMPACT OF THE STUDY: The beneficial effect of lowered fermentation pH to Lact. rhamnosus stability during storage in apple juice and the usefulness of malic acid test in predicting the stability were shown. PMID- 19191950 TI - Components derived from Pelargonium stimulate macrophage killing of Mycobacterium species. AB - AIMS: To determine the capacity of extracts of Pelargonium reniforme and Pelargonium sidoides, plants of the Geraniaceae family, to stimulate the uptake and killing of mycobacteria by murine macrophages and to identify the constituents that are responsible. METHODS AND RESULTS: Bioassay-guided fractionation of aqueous P. reniforme extracts yielded five chemically distinct structures with the capacity to increase the rate of intracellular killing by macrophages. These were: gallic acid, methyl gallate, myricetin and quercitin-3-O beta-d-glucoside, in addition to the previously unrecognized constituent 1-O-(2 (4-methoxyphenyl)ethyl-6-O-galloyl-glucopyranoside. Kinetics of intracellular accumulation of Mycobacterium tuberculosis and Mycobacterium fortuitum by macrophages were indistinguishable; pure preparations of the four previously known plant constituents stimulated macrophage killing, but not uptake, of M. tuberculosis and M. fortuitum equally well. CONCLUSIONS: A number of distinct molecular species are present in the medicinal plant P. reniforme that stimulate the killing of the intracellular pathogen M. tuberculosis. SIGNIFICANCE AND IMPACT OF THE STUDY: These observations support the view that Pelargonium extracts may have utility in the treatment of tuberculosis. PMID- 19191951 TI - The effect of surface charge, negative and bipolar ionization on the deposition of airborne bacteria. AB - AIMS: A series of experiments were conducted to evaluate the effect of surface charge and air ionization on the deposition of airborne bacteria. METHODS AND RESULTS: The interaction between surface electrostatic potential and the deposition of airborne bacteria in an indoor environment was investigated using settle plates charged with electric potentials of 0, +/-2.5kV and +/-5kV. Results showed that bacterial deposition on the plates increased proportionally with increased potential to over twice the gravitational sedimentation rate at +5kV. Experiments were repeated under similar conditions in the presence of either negative or bipolar air ionization. Bipolar air ionization resulted in reduction of bacterial deposition onto the charged surfaces to levels nearly equal to gravitational sedimentation. In contrast, diffusion charging appears to have occurred during negative air ionization, resulting in an even greater deposition onto the oppositely charged surface than observed without ionization. CONCLUSIONS: Static charges on fomitic surfaces may attract bacteria resulting in deposition in excess of that expected by gravitational sedimentation or simple diffusion. Implementation of bipolar ionization may result in reduction of bacterial deposition. SIGNIFICANCE AND IMPACT OF STUDY: Fomitic surfaces are important vehicles for the transmission of infectious organisms. This study has demonstrated a simple strategy for minimizing charge related deposition of bacteria on surfaces. PMID- 19191952 TI - Molecular typing and identification of Bdellovibrio-and-like organisms isolated from seawater shrimp ponds and adjacent coastal waters. AB - AIMS: To apply and compare two PCR-based methods for typing saltwater Bdellovibrio-and-like organisms (BALOs) and to understand ecological and phylogenetic aspects of the BALOs isolated from shrimp mariculture systems. METHODS AND RESULTS: Using double-layer agar technique, the numbers of culturable BALOs that lyse Vibrio alginolyticus were found to be 10-10(3) PFU ml(-1) in the surface water samples. A total of 130 BALOs isolates were differentiated into five phylotypes by denaturing gradient gel electrophoresis targeting the 16S rDNA V3 region and four phylotypes by amplified rDNA restriction analysis of the Bacteriovoracaceae-specific 16S rDNA fragment respectively. Phylogenetic analysis of representative isolates showed that all of them were identified as Bacteriovorax spp., but affiliated with four different clusters in the family Bacteriovoracaceae. CONCLUSIONS: The two PCR-based methods both can be chosen to rapidly type the saltwater BALOs at cluster level. And the relatively large numbers of BALOs with various phylotypes recovered from the same habitats suggested that these predators might play important ecological role in shrimp mariculture environments. SIGNIFICANCE AND IMPACT OF THE STUDY: We proposed two effective methods to distinguish rapidly large numbers of BALOs isolates and our results would be helpful to understand the diversity and function of BALOs in mariculture environments. PMID- 19191953 TI - Dairy bacteria remove in vitro dietary lectins with toxic effects on colonic cells. AB - AIMS: To assess in vitro the ability of some dairy bacteria to bind concanavalin A (Con A), peanut agglutinin (PNA) and jacalin (AIL), preventing their toxicity on mouse intestinal epithelial cells (IEC). METHODS AND RESULTS: Con A and AIL reduced significantly IEC viability in vitro, as determined by Trypan Blue dye exclusion or by propidium iodide/fluorescein diacetate/Hoescht staining. Different strains of dairy bacteria were able to remove lectins from the media. Two strains were subjected to treatments used to remove S-layer, cell wall proteins, polysaccharides and lectin-like adhesins. They were then assayed for the ability to bind dietary lectins and reduce toxicity against IEC and to adhere to IEC after interaction with lectins. Con A and AIL were removed by Propionibacterium acidipropionici and Propionibacterium freudenreichii by binding with specific sugar moieties on the bacterial surface. Removal of lectins by bacteria impaired IEC protection. Adhesion of P. acidipropionici to IEC was reduced but not abolished after binding Con A or AIL. CONCLUSIONS: Removal of Con A or AIL by dairy propionibacteria was effective to avoid the toxic effect against colonic cells in vitro. SIGNIFICANCE AND IMPACT OF THE STUDY: Consumption of foods containing these bacteria would be a tool to protect the intestinal epithelia. PMID- 19191954 TI - Atrazine degradation by stable mixed cultures enriched from agricultural soil and their characterization. AB - AIMS: The aim of this work was to enrich stable mixed cultures from atrazine contaminated soil. The cultures were examined for their atrazine biodegradation efficiencies in comparison with J14a, a known atrazine-degrading strain of Agrobacterium radiobacter. The cultures were also characterized to identify community structure and bacterial species present. METHODS AND RESULTS: The cultures were enriched and then stabilized in bacterial media. The stable mixed cultures and J14a were tested in a medium containing 100 microg l(-1) of atrazine. For all cultures, atrazine was removed 33-51% within 7 days and the cell optical density increased from 0.05 to between 0.50 and 0.70. Four isolates designated ND1, ND2, ND3 and ND4 were purified from the mixed cultures and identified based on sequence analysis of the 16 S rRNA gene as Alcaligenes faecalis, Klebsiella ornithinolytica, Bacillus megaterium and Agrobacterium tumefaciens, respectively. An atrazine-degrading gene, atzA, was present in ND2 and ND4. CONCLUSIONS: The stable mixed cultures obtained could degrade atrazine. Klebsiella ornithinolytica ND2 and Ag. tumefaciens ND4 are atrazine degraders. SIGNIFICANCE AND IMPACT OF THE STUDY: The novel stable mixed cultures could be used for bioremediating crop fields contaminated with atrazine. This is the first report of the atzA gene in Kl. ornithinolytica. PMID- 19191955 TI - Bacterial and archaeal assemblages in sediments of a large shallow freshwater lake, Lake Taihu, as revealed by denaturing gradient gel electrophoresis. AB - AIMS: To explore the association of microbial community structure with the development of eutrophication in a large shallow freshwater lake, Lake Taihu. METHODS AND RESULTS: The bacterial and archaeal assemblages in sediments of different lake areas were analysed using denaturing gradient gel electrophoresis (DGGE) of amplified 16S rDNA fragments. The bacterial DGGE profiles showed that eutrophied sites, grass-bottom areas and relatively clean sites with a eutrophic (albeit dredged) site are three respective clusters. Fifty-one dominant bacterial DGGE bands were detected and 92 corresponding clones were sequenced, most of which were affiliated with bacterial phylotypes commonly found in freshwater ecosystems. Actinobacteria were detected in the centre of the lake and not at eutrophied sites whereas the opposite was found with respect to Verrucomicrobiales. Twenty-five dominant archaeal DGGE bands were detected and 31 corresponding clones were sequenced, most of which were affiliated with freshwater archaeal phylotypes. CONCLUSIONS: The bacterial community structures in the sediments of different areas with similar water quality and situation tend to be similar in Taihu Lake. SIGNIFICANCE AND IMPACT OF THE STUDY: This study may expand our knowledge on the relationship between the overall microbial assemblages and the development of eutrophication in the shallow freshwater lake. PMID- 19191956 TI - Fermented soya bean (tempe) extracts reduce adhesion of enterotoxigenic Escherichia coli to intestinal epithelial cells. AB - AIMS: This study aimed to investigate the effect of processed soya bean, during the successive stages of tempe fermentation and different fermentation times, on adhesion of enterotoxigenic Escherichia coli (ETEC) K88 to intestinal brush border cells as well as Caco-2 intestinal epithelial cells; and to clarify the mechanism of action. METHODS AND RESULTS: Tempe was prepared at controlled laboratory scale using Rhizopus microsporus var. microsporus as the inoculum. Extracts of raw, soaked and cooked soya beans reduced ETEC adhesion to brush border cells by 40%. Tempe extracts reduced adhesion by 80% or more. ETEC adhesion to Caco-2 cells reduced by 50% in the presence of tempe extracts. ETEC K88 bacteria were found to interact with soya bean extracts, and this may contribute to the observed decrease of ETEC adhesion to intestinal epithelial cells. CONCLUSIONS: Fermented soya beans (tempe) reduce the adhesion of ETEC to intestinal epithelial cells of pig and human origin. This reduced adhesion is caused by an interaction between ETEC K88 bacteria and soya bean compounds. SIGNIFICANCE AND IMPACT OF THE STUDY: The results strengthen previous observations on the anti-diarrhoeal effect of tempe. This effect indicates that soya-derived compounds may reduce adhesion of ETEC to intestinal cells in pigs as well as in humans and prevent against diarrhoeal diseases. PMID- 19191957 TI - The improvement of amorpha-4,11-diene production by a yeast-conform variant. AB - AIMS: To investigate the effect of the yeast-conform variant of the Artemisia annua gene encoding for amorpha-4,11-diene synthase (ADS) on the production of amorpha-4,11-diene in a transformed yeast. METHODS AND RESULTS: The ADS gene was mutated to the yeast-conform variant ADSm. The ADSm synthesis was performed based on step-by-step extension of a short region of the gene through a series of polymerase chain reactions (PCR). The artificial ADSm gene contained codons preferred by the yeast translation machinery. The sequence was then integrated into a yeast expression vector pYeDP60. The fusion construct was active and the transformed yeast cells produced higher level of amorpha-4,11-diene compared with the plant gene-transformed yeast cells. CONCLUSIONS: Strains transformed with the yeast-conform allele (ADSm) were more efficient in terms of production of amorpha 4,11-diene than those transformed with the plant gene. SIGNIFICANCE AND IMPACT OF THE STUDY: We demonstrated that yeast-conform allele of foreign genes by serial PCR reactions can be a solution to low efficiency of heterologous gene expression in Saccharomyces cerevisiae cells. PMID- 19191958 TI - Acrebol, a novel toxic peptaibol produced by an Acremonium exuviarum indoor isolate. AB - AIMS: To identify a toxin and its producer isolated from woody material in a building where the occupants experienced serious ill health symptoms. METHODS AND RESULTS: Hyphal extracts of an indoor fungus, identified as the cycloheximide tolerant species Acremonium exuviarum, inhibited motility of boar spermatozoa (EC(50) 5 +/- 2 microg of crude solids ml(-1)) and caused cytolysis of murine neuroblastoma cells (MNA) and feline fetal lung cells (FL). The responsible substances were purified and identified as two structurally similar, heat-stable, novel, toxic peptaibols, 1726 Da and 1740 Da, respectively, with amino acid sequences of Acetyl-Phe-Iva/Val-Gln-Aib-Ile-Thr-Leu-Aib-Pro-Aib-Gln-Pro-Aib-(X-X X)-SerOH and Acetyl-Phe-Iva/Val-Gln-Aib-Ile-Thr-Leu-Val-Pro-Aib-Gln-Pro-Aib-(X-X X)-SerOH. Purified acrebol inhibited motility of boar sperm, depleted ATP half content in 1 day (EC(50) of 0.1 microg ml(-1), 60 nmol l(-1)) depolarised the mitochondria after 2 days, but did not affect the cellular content in NADH. This indicates mitochondrial toxicity. Plate-grown biomass of A. exuviarum BMB4 contained 0.1-1% (w/w) of acrebol, depending on the culture medium. CONCLUSIONS: Acrebol paralysed the energy generation of mammalian cells suggesting that mitochondria were its target of action. SIGNIFICANCE AND IMPACT OF THE STUDY: Acremonium exuviarum, as an indoor fungus, is potentially hazardous to health because of the toxic peptaibols that it produces. PMID- 19191959 TI - Protective effect of salicylates against hydrogen peroxide stress in yeast. AB - AIMS: To investigate the effects of salicylates in Saccharomyces cerevisiae exposed to oxidative stress induced by hydrogen peroxide (H(2)O(2)). METHODS AND RESULTS: Saccharomyces cerevisiae was cultured through to the postlogarithmic phase of growth. Stress was induced by the addition of 1.5 mmol l(-1) H(2)O(2) for 1 h, while N-acetyl-l-cysteine (NAC) and glutathione (GSSG) were used as control agents that affect the redox balance. Sodium salicylate, at 0.01-10 mmol l(-1)or acetylsalicylic acid, at 0.02-2.5 mmol l(-1) was administered at various times before hydrogen peroxide stress. Both agents conferred resistance to a subsequent hydrogen peroxide stress, similarly to the induction of the adaptive response observed upon pretreatment with NAC and GSSG. Sodium salicylate was more potent as a short-term, but not as a long-term pretreatment agent, compared to acetylsalicylic acid. CONCLUSIONS: Pharmacological pretreatment with salicylates resulted in dose related increases in cell survival, indicating the induction of the protective response in yeast. SIGNIFICANCE AND IMPACT OF THE STUDY: The possible role of salicylates in the modulation of the hydrogen peroxide stress response in eukaryotic cells address questions on the effects of these commonly used therapeutic agents in a number of disorders exhibiting an oxidative stress component. PMID- 19191960 TI - A unique lantibiotic, thermophilin 1277, containing a disulfide bridge and two thioether bridges. AB - AIMS: To identify the chemical structure of a bacteriocin, thermophilin 1277, produced by Streptococcus thermophilus SBT1277. METHODS AND RESULTS: Thermophilin 1277 was purified and partial N-terminal sequence analysis revealed 6 unidentified amino acids amongst 31 amino acids residues. A 2.7-kbp region containing the thermophilin 1277 structural gene (tepA) encoding 58 amino acids was cloned and sequenced. Mature thermophilin 1277 (33 amino acids) was preceded by a 25-amino acid putative leader peptide containing a double glycine cleavage motif. Peptide sequence analysis following chemical modification of thermophilin 1277 revealed that the Cys21 and Cys29 residues form a disulfide bridge and that Thr8 or Thr10 forms two 3-methyllanthionines with Cys13 or Cys32 via thioether bridges. Antimicrobial activity was disrupted by ethanethiol or reductive agent treatments, indicating that the internal amino acid modifications are crucial for the activity. CONCLUSIONS: Thermophilin 1277 from Strep. thermophilus SBT1277 belongs to the class of AII-type lantibiotics that has a disulfide and two thioether bridges. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of a lantibiotic produced by a GRAS species of Strep. thermophilus; thermophilin 1277 has a unique structure containing both a disulfide bridge and two thioether bridges that are crucial for its activity. PMID- 19191961 TI - Delineation of Pseudomonas savastanoi pv. savastanoi strains isolated in Tunisia by random-amplified polymorphic DNA analysis. AB - AIMS: To investigate the genetic diversity of Pseudomonas savastanoi pv. savastanoi strains and to look whether these strains were distributed to geographical location. METHODS AND RESULTS: Random amplification of polymorphic DNA (RAPD) was used to discriminate between 58 Tunisian strains and 21 strains from various other countries of P. savastanoi pv. savastanoi, the causal agent of olive knot disease. Isolates were separated into three groups by cluster analysis and principal coordinate analysis of RAPD fingerprint data obtained with three primers (OPR-12, OPX-7 and OPX-14). Group 1 contained isolates from the southeast of Tunisia and European strains. Group 2 comprised strains isolated from the north of Tunisia exclusively while group 3 encompassed the majority of isolates obtained from five orchards located in the centre of Tunisia. CONCLUSIONS: The results indicated that isolates of P. savastanoi pv. savastanoi were genetically distinct according to geographic regions. RAPD grouped isolates derived from the same orchard as identical. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first application of RAPD in the delineation of P. savastanoi pv. savastanoi strains. PMID- 19191962 TI - Inhibitory effects of sucrose fatty acid esters on survival of Yersinia enterocolitica on eggshell surface and use of blue lake as indicator of bacterial penetration into eggs. AB - AIMS: To determine the effectiveness of sucrose monolaurate (SML) and sucrose monocaprate (SMC), alone and in combination with ethylenediaminetetraacetic acid (EDTA), propionic acid (PA) or citric acid (CA) in reducing mesophilic aerobic bacteria (MAB) and Yersinia enterocolitica O:9 populations on eggshells and their damage potential on the microstructure of shell cuticle. METHODS AND RESULTS: Uninoculated eggs and eggs submerged in a solution of Y. enterocolitica were immersed in solutions of the various treatments. MAB and Y. enterocolitica counts on the surface of the eggs were carried out before and after treatment. MAB counts decreased less than 2 logs on uninoculated eggshells irrespective of treatment and reductions of 3.2 and 3.0 logs of Y. enterocolitica were obtained with 1000 microg ml(-1) SML plus 0.1% CA or 1000 microg ml(-1) SML plus 600 microg ml(-1) EDTA solutions, respectively. Y. enterocolitica 2/O:9 was recovered from natural microflora. Use of blue lake staining revealed minimal damage to the shells from the washing treatments. CONCLUSIONS: SML and SMC at 1000 microg ml( 1) combined with CA or EDTA could be effective in reducing Y. enterocolitica on eggshells with a minimal risk of later bacterial recontamination. SIGNIFICANCE AND IMPACT OF THE STUDY: Eggs are a recognized vehicle for transmission of Y enterocolitica although a prevalence of only 2.7% was detected in this study. Washing eggs in solutions containing SML or SMC could eliminate Y. enterocolitica contamination of egg shells. PMID- 19191963 TI - Effects of dietary probiotic supplementations on prevention/treatment of yersiniosis disease. AB - AIMS: To evaluate Enterobacter cloacae and Bacillus mojavensis, isolated from rainbow trout gut in the present study, as a probiotic to control yersiniosis disease. METHODS AND RESULTS: A strain of Ent. cloacae and B. mojavensis, isolated from the digestive tract of rainbow trout had an antagonistic effect to Yersinia ruckeri, which causes yersiniosis. After feeding fish with 1 x 10(8) cells g(-1) probiotic containing feed for 60 days, the fish survival rate increased to 99.2% following challenge with Y. ruckeri compared with controls that had 35% survival rate. Effects of Ent. cloacae and B. mojavensis on weight gains and stimulation of red blood cells, white blood cells, platelets and hemoglobin were also evaluated in separate groups of fish fed probiotic containing feed for 2 months. Probiotic significantly affected white blood cells, hemoglobin and weight gains of the experimental fish. CONCLUSIONS: Enterobacter cloacae and B. mojavensis, can be used to prevent and control yersiniosis disease. SIGNIFICANCE AND IMPACT OF THE STUDY: In conclusion, concomitant use of Ent. cloacae and B. mojavensis as a feed supplement is beneficial to rainbow trout. Use of these organisms can protect fish from yersiniosis and enhance digestibility and utilization of feed. Use of such probiotics may also limit the use of antibiotics and other chemicals in control and treatment of diseases, and thus contribute to the effort to reduce environmental contamination by residual antibiotics and chemicals. PMID- 19191964 TI - Biochemical characteristics and function of a threonine dehydrogenase encoded by ste11 in Ebosin biosynthesis of Streptomyces sp. 139. AB - AIMS: Ebosin, a novel exopolysaccharide (EPS) produced by Streptomyces sp. 139 has antagonistic activity for interleukin-1 receptor (IL-1R) in vitro and remarkable anti-rheumatic arthritis activity in vivo. Ebosin biosynthesis gene (ste) cluster has been identified in our laboratory. This paper reports our effort to characterize the function of ste11 gene. METHODS AND RESULTS: After the ste11 gene was cloned and expressed in Escherichia coli BL21, the recombinant Ste11 was purified and found capable of catalyzing NAD(+) and l-threonine to NADH and 2-amino-3-ketobutyrate, hence identified as a threonine dehydrogenase (TDH). To investigate its function in the biosynthesis of Ebosin, the ste11 gene was knocked out with a double crossover via homologous recombination. The monosaccharide composition of EPS produced by the mutant strain (EPS-m) was altered from that of Ebosin. The analysis of IL-1R antagonist activity for EPSs showed that the bioactivity of EPS-m was lower than Ebosin. CONCLUSIONS: ste11 gene encoding a TDH may function as a modifier gene of Ebosin during its biosynthesis. SIGNIFICANCE AND IMPACT OF THE STUDY: TDH encoded by ste11 is functional in Ebosin biosynthesis. It is the first characterized TDH in Streptomyces. PMID- 19191965 TI - Evaluation of the 'GeneDisc' real-time PCR system for detection of enterohaemorrhagic Escherichia coli (EHEC) O26, O103, O111, O145 and O157 strains according to their virulence markers and their O- and H-antigen-associated genes. AB - AIMS: To evaluate the GeneDisc multiplex real-time PCR assay for detection of enterohaemorrhagic Escherichia coli (EHEC) O26, O103, O111, O145 and O157 strains. METHODS AND RESULTS: GeneDiscs for detection of genes encoding Shiga toxins (stx), intimins (eae), E. coli O157 (rfbE(O157)) and H7 (fliC(H7)) antigens as well as genes specific for EHEC O26 (wzx(O26)), O103 (wzx(O103)), O111 (wbd1(O111)), O145 (ihp1(O145)) and O157 (ihp1(O157)) were evaluated. The assay was run with native bacteria in 1 h in a GeneDisc Cycler. All genotypes of stx and eae, except stx(2f) and eae-rho, were identified. Escherichia coli strains belonging to O-groups O26, O103, O111, O157 as well as EHEC O145:[H28] strains were specifically detected with this assay. The ihp1(O157) gene was not found specific for EHEC O157. O-rough mutants of EHEC and non-motile EHEC O157 strains were reliably identified with the GeneDisc assay. Two to three colonies of EHEC strains were still detectable in a lawn of 50 000 apathogenic E. coli from agar plates. CONCLUSIONS: The GeneDisc assay is a specific and reliable assay for detection of major EHEC strains. It is robust enough to detect few EHEC colonies in mixed cultures of bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: The assay is promising for its use in EHEC diagnostics and for EHEC monitoring with different kinds of samples. PMID- 19191966 TI - New insights into the biodegradation of thiodiglycol, the hydrolysis product of Yperite (sulfur mustard gas). AB - AIMS: To isolate thiodiglycol (TDG)-degrading bacteria, the mustard gas hydrolysis product, and to characterize the metabolites formed and the enzymes involved in the degradation. METHODS AND RESULTS: Two strains, identified as Achromobacter xylosoxydans G5 and Paracoccus denitrificans E4, isolated from a petroleum-contaminated soil, utilized TDG as sole carbon and sulfur source. During the degradation of TDG by strain E4 [(2-hydroxyethyl)thio] acetic acid (HETA), thiodiglycolic acid (TDGA) and bis-(2-hydroxyethyl)disulfide (BHEDS) were identified by gas chromatography-mass spectrometry analysis, while HETA and TDGA were identified for strain G5. Two-dimensional isoelectric focussing-gel electrophoresis (2-D IEF/SDS-PAGE) maps of protein extracts of P. denitrificans E4 grown on TDG showed a spot identified as a methanol dehydrogenase. Increased expression of a putative iscS gene, involved in sulfur assimilation, was observed in TDG-grown cells of A. xylosoxydans G5. CONCLUSIONS: TDG degradation by P. denitrificans E4 occurred through two pathways: one involved cleavage of the C-S bond of HETA, yielding BHEDS and the other, oxidation of the alcoholic groups of TDG, yielding TDGA. The cleavage of the C-S bond of TDGA gave mercaptoacetic acid, further oxidized to acetate and sulfate. SIGNIFICANCE AND IMPACT OF THE STUDY: Increased knowledge of TDG-degrading bacteria and the possibility of using them in a tailored-two-stage mustard gas destruction process. PMID- 19191967 TI - Campylobacter jejuni: a study on environmental conditions affecting culturability and in vitro adhesion/invasion. AB - AIMS: Nongrowing cultures of Campylobacter jejuni lose their culturability. It is unclear whether this loss in culturability also affects their ability to interact with host cells. The purpose of this study was to determine the relevance of the number of culturable cells to the ability to adhere/invade in Caco-2 cells. METHODS AND RESULTS: For C. jejuni C356, culturability and adhesion/invasion were monitored in time (days) under different storage conditions (temperature, medium, atmosphere). Decrease rates of both culturability and adhesion/invasion were dependent on the conditions used, but the number of adhering/invading cells per culturable cell was not affected by the environmental conditions. Furthermore five strains were monitored at one condition. The culturability and adhesion/invasion decrease rates did not significantly differ per strain; however the number of adhering/invading cells per culturable cell was strain dependent. CONCLUSIONS: Culturability and adhesion/invasion of C. jejuni are linearly related. The number of adhering/invading cells per culturable C. jejuni cell is strain dependent, but is not affected by environmental conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: It was shown that the number of culturable cells is a good measure for the in vitro adhesion/invasion of. C. jejuni. PMID- 19191968 TI - Effect of gamma-irradiation on membrane fatty acids and peptidoglycan's muropeptides of Pantoea agglomerans, a plant pathogen. AB - AIMS: The aim of this study was to evaluate the effect of gamma-irradiation on the fatty acids (FA) and muropeptides content of two strains of an Enterobacteriacea: Pantoea agglomerans. METHODS AND RESULTS: Pantoea agglomerans strains ATCC 49174 and RL1 isolated from irradiated carrots were used for this study. Radiation treatments (1 and 3.5 kGy) were performed to study the radiotolerance. Total lipids were obtained by multiple extractions using methanol/chloroform (2 : 1) and were quantified by GC. Muropeptides were purified by successive enzymatic digestions and analysed using a reverse phase C(18) column in high performance liquid chromatography. A significant (P or= 37.8 degrees C). The mean bias between rectal and ear temperatures as measured with the ThermoScan was -0.20 degrees C (SD 0.32) degrees C and the 95% limits of agreement were -0.83 degrees C and 0.42 degrees C (95% CI, -0.88-0.48 degrees C). Using the Genius, the corresponding figures were 0.56 degrees C (SD 0.39) degrees C, -1.32 degrees C and 0.20 degrees C (95% CI, 1.39-0.27 degrees C). After correction for bias, the ThermoScan predicted the level of fever with a maximum error of 0.7 degrees C (mean error 0.3 degrees C). Using the Genius, the maximum error and the mean error were 1.6 degrees C and 0.4 degrees C, respectively. CONCLUSIONS: Infrared ear thermometry can predict rectal temperature in normothermic and in febrile inpatients with an acceptable level of accuracy. However, the predictive accuracy depends on both operator technique and quality of instrumentation. RELEVANCE TO CLINICAL PRACTICE: Proper technique (measuring in both ears) and optimal instrumentation (model of ear thermometer) are essential for accuracy. PMID- 19191994 TI - Commentary on McCance T, Slater P & McCormack B (2009) Using the caring dimensions inventory as an indicator of person-centred nursing. Journal of Clinical Nursing, 18, 409-417. PMID- 19191996 TI - Editorial: Nursing amidst stories (and knowing what to do with them). PMID- 19191997 TI - A review of the information and support needs of family carers of patients with chronic obstructive pulmonary disease. AB - AIMS AND OBJECTIVES: The objectives of this narrative review were to identify: (1) The information and support needs of carers of family members with chronic obstructive pulmonary disease; (2) appropriate interventions to support carers in their caregiving role; (3) information on carers' needs as reported in studies of patients living with COPD in the community. BACKGROUND: Chronic obstructive pulmonary disease is a major health problem in the UK resulting in significant burden for patients, families and the health service. Current National Health Service policies emphasise, where medically appropriate, early discharge for acute exacerbations, hospital-at-home care and other models of community care to prevent or reduce re-hospitalisations of people with chronic conditions. Understanding carers' needs is important if health care professionals are to support carers in their caregiving role. DESIGN: A narrative literature review. METHODS: Thirty five papers were reviewed after searching electronic databases. RESULTS: Few studies were identified which addressed, even peripherally, carers' needs for information and support, and no studies were found which described and evaluated interventions designed to enhance caregiving capacity. Several studies of hospital-at-home/early discharge, self care and home management programmes were identified which included some information on patients' living arrangements or marital status. However, there was little or no detail reported on the needs of, and in many cases, even the presence of a family carer. CONCLUSIONS: This review highlights the dearth of information on the needs of carers of chronic obstructive pulmonary disease patients and the need for future research. RELEVANCE TO CLINICAL PRACTICE: There is little research based knowledge of the needs of carers of chronic obstructive pulmonary disease patients and interventions to assist them in providing care. This knowledge is critical to ensure that carers receive the information they need to carry out this role while maintaining their own physical and emotional health. PMID- 19191998 TI - Informal carers of older family members: how they manage and what support they receive from respite care. AB - AIMS: To describe informal carers' coping strategies and their experiences of the support provided by respite care. BACKGROUND: The target set in the Finnish national old age policy strategy is that 90% of the population over 75 should be able to live at home. A major source of support for older people and their families is respite care in an institution. DESIGN: Survey. METHODS: The data were collected among informal carers who regularly use respite care services (n = 143). Two structured questionnaires were used: the Carers' Assessment of Managing Index (Nolan et al. 1995) and an index developed by the researchers. RESULTS: Informal carers who provided care for younger individuals or people who needed less help had a better quality of life. Spouse carers and older informal carers were less satisfied with their quality of life than younger carers. Periods of respite care had a major influence on informal carers' quality of life: 93% said they felt invigorated. Care periods remained rather isolated episodes in the lives of the older patients in that there was hardly any discussion at the institution about the family's situation, the objectives of respite care, or about how the informal carer was coping. Altogether, 47% of the nurses remained fairly distant. The most useful coping strategies involved creating alternative perspectives in the caring relationship. CONCLUSIONS: The results strengthen the knowledge base of informal caregiving by showing how caregivers cope and what kind of support they need in their demanding caring role. RELEVANCE TO CLINICAL PRACTICE: Respite care nurses should invest greater effort in supporting informal carers. The service should be developed as an integral part of home care for older people. Nurses and informal carers should see each other as partners who share a common goal, i.e. the well-being of the entire family. PMID- 19191999 TI - Stressors, social support, depressive symptoms and general health status of Taiwanese caregivers of persons with stroke or Alzheimer's disease. AB - AIMS AND OBJECTIVES: This study examined the relationships among stressors, social support, depressive symptoms and the general health status of Taiwanese caregivers of individuals with stroke or Alzheimer's disease. BACKGROUND: Caring for a disabled or cognitively impaired person can be extremely stressful and often has adverse effects on caregivers' health. While research on caregiving in Taiwan has examined caregivers' characteristics, caregivers' need and caregivers' burden in caring for older people in general, little is known about Taiwanese caregivers of individuals with stroke or Alzheimer's disease. DESIGN: Cross sectional, descriptive correlation design. METHODS: Data were obtained from a convenience sample of 103 Taiwanese informal caregivers in the South of Taiwan and analysed using descriptive statistics, Pearson's correlations, multiple and hierarchical regressions and t-tests. RESULTS: Caregivers who had lower household incomes and were taking care of individuals with more behaviour problems had more depressive symptoms. In addition, caregivers who were older and were taking care of individuals with more behaviour problems had worse general health. Caregivers who had more emotional support had less depressive symptoms. CONCLUSIONS: Caregivers of persons with Alzheimer's disease had more depressive symptoms and worse general health than caregivers of persons with stroke. Only emotional support moderated the relationship between one of the stressors (household income) and depressive symptoms. RELEVANCE TO CLINICAL PRACTICE: The findings of this study may be helpful for nurses and other health care professionals in designing effective interventions to minimise the negative impacts of stressors on the psychological and general health of caregivers in Taiwan. PMID- 19192000 TI - Assuring the integrity of the family: being the father of a very low birth weight infant. AB - AIM: The purpose of this study was to explore the paternal role of fathers of very low birth weight infants during their first year of life. BACKGROUND: Following birth, very low birth weight infants often require extended hospitalisation and long-term care. The birth experience and the infant's hospitalisation often overwhelm parents and may hinder the development of their parental roles. The father is known to be an important supporting figure for the mother, but most studies in Taiwan have focussed on mothers. DESIGN: A qualitative design using a grounded theory approach was employed in this study. METHODS: In-depth interviews were conducted with a purposive sample of fathers during home visits. Each father was interviewed twice at one to three months and seven to nine months after his very low birth weight infant's discharge from the hospital. Data were analysed using constant comparison and repeated verification. RESULTS: Twelve fathers were included in the study. During the first year of life of their very low birth weight infants, the fathers experienced 'protecting the wife-baby dyad', 'concentrating on the child's health and growth', and 'possessing a complete family'. Subcategories within each major category also emerged. CONCLUSION: In this study, Taiwanese fathers of very low birth weight infants learned different and broader dimensions of the paternal role and continually made adjustments to assure their family's integrity. RELEVANCE TO CLINICAL PRACTICE: These findings can help nurses understand paternal perceptions and behaviours related to having a very low birth weight infant and in turn help fathers adjust to their role. PMID- 19192001 TI - The dramatic encounter: experiences of taking part in a health conversation. AB - AIM: The purpose of this study was to describe lived experiences of a health conversation from the perspective of participants who, in the course of a health check, had been informed that they were at increased risk of cardiovascular heart disease. BACKGROUND: The modern lifestyle has created an increased occurrence of cardiovascular heart disease. Counselling about lifestyle changes to prevent disease is an important duty of nurses in primary healthcare when encountering patients with risk factors such as diabetes, hypertension and hypercholesterolemia. It is well known that accomplishing lifestyle changes is complicated. Research has shown the importance of assuming the patient perspective in pedagogical encounters and that nurses involved in patient teaching need more knowledge about the learning person. DESIGN: Written narratives about the health conversation followed by interviews with open-ended questions. Nine informants from the Skaraborg Project participated. METHODS: A phenomenological analysis was chosen to describe the participants' experiences and to visualise the meaning of the new knowledge obtained in the encounter. RESULTS: Three themes were identified; 'The unavoidable message', 'Reflection on the content of the conversation' and 'The pedagogical encounter'. Sub-themes show the variations of meaning owing to the participants' previous life experiences. CONCLUSIONS: The nurse must be aware of differences in perceptions concerning the health conversation and possible consequences for the life situation. The health conversation encounter requires preparedness and a pedagogical awareness as well as an ability to recognise the person's need for understanding and level of motivation to make lifestyle changes. Relevance to clinical practice. Time and space must be created for health conversations and follow-up of cardiovascular heart disease to achieve a preventive pedagogical encounter in primary healthcare. PMID- 19192002 TI - Disease knowledge and treatment adherence among patients with thalassemia major and their mothers in Taiwan. AB - AIMS: The current study had three aims: (i) to examine disease knowledge in both thalassemia major patients and their mothers; (ii) to understand the relationships between disease knowledge and treatment adherence in thalassemia major patients; and (iii) to explore the importance of selected factors in predicting patients' knowledge about thalassemia major. BACKGROUND: Patients with thalassemia major must be treated with life-long blood transfusions. Evidence suggests that patients with more knowledge/information about their illnesses adhere more readily to treatment schedules. However, there has been little evaluation of knowledge and treatment adherence in thalassemia major patients. DESIGN AND METHODS: A cross-sectional correlational survey design and purposive sampling were used. Thirty-two thalassemia major patients (mean age 17.5 years) and 32 mothers (mean age 40.5 years) were recruited. RESULTS: On a scale ranging from 0-20, the average of the patients' disease knowledge about thalassemia major was 15.19 and the average of their mothers' disease knowledge was 16.44. The scores for the patients' disease knowledge about thalassemia major were positively correlated with follow-up visit adherence (r = 0.690, p < 0.001) and with desferrioxamine infusion adherence (r = 0.791, p < 0.001). 95.6% of variance in patients' knowledge was explained by a model that included mothers' knowledge (beta = 0.901, p < 0.001), follow-up visit adherence (beta = 0.084, p = 0.140) and annual household income (beta = 0.042, p < 0.387). CONCLUSIONS: The positive association between knowledge and treatment adherence and factors of patients' knowledge indicate the need for systematic education for patients and caregivers to improve adherence to treatment. RELEVANCE TO CLINICAL PRACTICE: Improvement of the quality of patient care, reinforcement of medical education and enhanced efforts by clinical staff to provide practical knowledge to patients with thalassemia major should significantly improve patient adherence to treatment. PMID- 19192003 TI - The relationship between caregiver's strain and social support among mothers with intellectually disabled children. AB - AIMS AND OBJECTIVES: To explore caregiver's strain, the relationship between social support and caregiver's strain and the predictors of caregiver's strain among mothers with school-aged intellectually disabled children in Taiwan. BACKGROUND: Strain is a common condition among mothers who take care of intellectually disabled children; this correspondingly reduces their ability to care for children, thereby affecting the functioning of the entire family. Yet, there have been very few studies on caregivers of intellectually disabled children of school age. DESIGN: Cross-section correlational design. METHODS: Data collection consisted of face-to-face interviews combined with a structured questionnaire. Instruments employed were the Caregiver Strain Index, Social Support Scale and three open-ended questions. In total, 127 mothers completed the questionnaire. RESULTS: Results showed that mothers with intellectually disabled children had a rather high level of strain and received inadequate social support. Social support and strain had a significant and negative correlation. Stepwise regression analysis revealed that mothers' health status, social support and amount of time spent as a caregiver, as well as the intellectually disabled children's dependent degree of daily living activity, were major predictors of caregiver's strain, which accounted for 38.4% of the total variance. CONCLUSIONS: The results provide a guide for healthcare professionals in designing effective interventions and preventive care to reduce the level of strain in mothers with intellectually disabled children. This, in turn, could improve the quality of life of the mother and her family. RELEVANCE TO CLINICAL PRACTICE: In Taiwan, care of intellectually disabled children is primarily provided by family members. Therefore, we should emphasise family-centred care to enable healthcare professionals to become more effective as case managers in local clinics, schools and communities. PMID- 19192004 TI - Systems and people under pressure: the discharge process in an acute hospital. AB - AIMS: To understand the perspective of hospital-based health professionals with regard to preparing patients for discharge from an acute hospital in England. BACKGROUND: The hospital experience in England over recent years is characterised by increasing admission rates and decreasing length of stay. Legislation and policy initiatives have also focussed upon the need to reduce delayed discharges. Discharge preparation is known to be a complex intervention with multiple obstacles within and outside of the hospital setting. DESIGN: Qualitative. METHODS: Posters were displayed within a hospital asking health professionals to take part in a focus group. Maximum variation, in terms of job titles, was sought for within the sample. Focus groups were held in December 2006. Six senior members of staff divided into pairs to run them. All groups were taped and transcribed verbatim and analysed using a framework approach. RESULTS: Three focus groups were conducted, which involved 11 nurses, 15 allied health professionals, five social workers and one doctor. Analysis identified the following themes and sub themes: 1 Conflicting pressures on staff: Keeping patients in hospital vs. getting them out; Striving for flexibility within a system; A paucity of intermediary provision. 2 Casualties arising from conflicting pressures: Professionals losing their sense of professionalism; Patients being 'systematised'. CONCLUSIONS: Pressures described during focus groups stemmed from five main sources: external targets placed upon the system, internal hospital inflexibility and poor communication, the dominance of the medical model of care, a desire to address the complex needs of individuals and a lack of community services. Staff felt themselves to be victims of these competing pressures and that many of the solutions were beyond their influence. Staff described the dehumanising effect of sometimes having to ignore patient concerns, wishes and choices. RELEVANCE TO CLINICAL PRACTICE: Understanding of the pressures surrounding discharge could inform relevant service improvements. PMID- 19192005 TI - Perceived adverse patient outcomes correlated to nurses' workload in medical and surgical wards of selected hospitals in Kuwait. AB - AIM: This study was carried out to identify the perceived adverse patient outcomes as related to nurses' workload. It also assessed nurses' perception of variables contributing to the workload and adverse patient outcomes. BACKGROUND: Several studies have been published on adverse patient outcomes in which a correlation was found between nurses' workload and some adverse patient outcomes. DESIGN: A cross-sectional survey was conducted between registered nurses (n = 780) working in medical and surgical wards of five general governmental hospitals in Kuwait. DATA COLLECTION INSTRUMENTS: Data were collected using a self administered questionnaire consisting of three sections to elicit information about the sample characteristics, perception of workload and perceived adverse patient outcomes during the last shift and last working week. RESULTS: The three major perceived adverse outcomes reported by the nurses while on duty during their last shift were: complaints from patients and families (2%), patients received a late dose or missed a dose of medication (1.8%) and occurrences of pressure ulcer (1.5%). Similarly, the reported adverse outcomes over the past week were complaints from patients and families (5%), patients received a late dose or missed a dose of medication (5.3%) and discovery of a urinary tract infection (3.7%). Increases in nurse-patient load, bed occupancy rate, unstable patients' condition, extra ordinary life support efforts and non-nursing tasks; all correlated positively with perceived adverse patient outcomes. CONCLUSION: This study sheds light on an important issue affecting patient safety and quality of care as perceived by the nurses themselves as caregivers. RELEVANCE TO CLINICAL PRACTICE: Nurses' perception of variables contributing to adverse patient outcomes and their workload could significantly affect the provided nursing care and nursing care recipients. The findings could help in policy formulation and planning strategies to decrease adverse patient outcomes in many countries with a health care structure similar to that of Kuwait. PMID- 19192006 TI - HIV prevention for people with serious mental illness: a survey of mental health workers' attitudes, knowledge and practice. AB - AIM: The aim of this survey was to investigate the attitudes, knowledge and reported practice (capabilities) of mental health workers concerning human immunodeficiency virus (HIV) and other sexually transmitted diseases in people with serious mental illness. BACKGROUND: People with serious mental illness are at increased risk of HIV and other sexually transmitted infections. Mental health workers have a key role to play in promoting sexual health in this population, but it is unclear how they perceive their role in this work and whether they have the capabilities to deliver sexual health promotion. DESIGN: Cross sectional survey. METHODS: A questionnaire was devised and distributed to 650 mental health workers working in a London (UK) NHS mental health service. RESULTS: A response rate of 44% was achieved. Overall, workers reported positive attitudes to sexual health promotion and were knowledgeable about risk behaviours and risk factors for HIV infection. Adherence to glove wearing was good. However, participants' knowledge about HIV/AIDS in people with schizophrenia was poor and most reported they were not engaged in sexual health promotion activities with people with serious mental illness. Glove wearing was predicted by those who had drug and alcohol training and clinical experience and knowledge of risk factors was predicted by previous health promotion training. No other demographic factors predicted any of the other subscales. CONCLUSION: Mental health workers require training to provide skills for health promotion regarding sexual health and HIV in people with serious mental health problems. In addition, there needs to be more research on risk behaviours. RELEVANCE TO CLINICAL PRACTICE: The development of effective interventions to reduce this behaviour. PMID- 19192008 TI - Commentary on Taylor J, Baldwin N and Spencer N (2008) 'Predicting child abuse and neglect; ethical, theoretical and methodological challenges'. Journal of Clinical Nursing 17, 1193-1200. PMID- 19192009 TI - Response to Carter, B (2008) Commentary on Shields L, Pratt J, Hunter J (2006) Family centred care: a review of qualitative studies Journal of Clinical Nursing 15, 1317-1323 in Journal of Clinical Nursing 17, 2091-2093. PMID- 19192011 TI - De novo tuberculosis during infliximab therapy in a patient with Behcet disease. AB - A woman with oculocutaneous Behcet disease developed primary tuberculosis while being treated with infliximab. A latent tuberculosis infection had been excluded before therapy. After more than 80 weeks of treatment, the patient complained of fevers, night sweats, shivering and vigorous cough. The chest x-ray showed miliary shadowing. Mycobacterium tuberculosis was identified. The history revealed recent contact to an individual with smear-positive tuberculosis. This constellation speaks in favor of a de novo tuberculosis infection with a fulminant course. PMID- 19192012 TI - Trichoblastomas with Merkel cell proliferation in nevi sebacei in Schimmelpenning Feuerstein-Mims syndrome--histological differentiation between trichoblastomas and basal cell carcinomas. AB - The hallmark of Schimmelpenning-Feuerstein-Mims syndrome (SFMS) is a systematized nevus sebaceous that follows Blaschko lines and usually involves the face. It represents a rare congenital nevus syndrome with alterations of skin, bones, CNS, eyes and heart. Nevi sebacei can proliferate and develop into epithelial tumors like trichoblastoma, syringocystadenoma and basal cell carcinoma. The histological differentiation between basal cell carcinoma and trichoblastoma is difficult. We present an adult woman with SFMS who was followed by multiple specialties since birth without the correct diagnosis being made. She was referred to us with the diagnosis of multiple basal cell carcinomas of head and face. Our diagnosis of systematized nevus sebaceus was crucial for the correct classification of SFMS. We identified multiple trichoblastomas in the nevi sebacei and could exclude basal cell carcinomas. The essential clue was the detection of multiple Merkel cells within the epidermal layer by cytokeratin 20 staining. PMID- 19192013 TI - Tamoxifen-induced melasma in a postmenopausal woman. PMID- 19192014 TI - Paediatric molluscum folliculitis: a diagnosis to keep in mind. PMID- 19192015 TI - Interest of the association retinaldehyde/glycolic acid in adult acne. AB - BACKGROUND: Topical retinoids have been successfully used in the treatment of acne vulgaris but may induce irritation when used twice daily. The association of retinaldehyde (RAL) with glycolic acid (GA) have complementary activities, which could be of interest for adult women with acne because of a better tolerance/efficacy ratio. The aim of this study was to evaluate the tolerance and the efficiency of RAL (0.1%)/GA (6%) in adult women with acne when used alone or in combination with their usual acne products except retinoids. METHODS: Three hundred ninety-seven women with acne (aged between 30 and 40 years old) were included in this open multicentric study. They had to apply cream containing RAL/GA for 90 days without stopping their previous acne treatment (except topical retinoids). The tolerance was the main criteria and the second one is the efficacy, which was assessed by counting inflammatory and retentional lesions after 30 and 90 days of treatment. RESULTS: Used alone or in association with other anti-acne treatments, RAL/GA was considered to be highly tolerated. A significant decrease in both inflammatory and retentional lesions between day 0 and day 90 indicates that RAL/GA can be used as monotherapy for mild acne or could potentate the efficiency of other anti-acne products used at the same time by patients suffering from moderate acne. Complaints about side-effects were rare. The subjective evaluation of the preparation's efficacy by investigators and patients was strongly favourable. CONCLUSION: These data show that a combination of RAL 0.1% and GA 6% may be used in association with other topical anti-acne treatments with an excellent tolerance. PMID- 19192016 TI - Double-blind, randomized, placebo-controlled trial of a cream containing the Stryphnodendron adstringens (Martius) Coville bark extract for suppressing terminal hair growth. AB - BACKGROUND: Excess of terminal hair can be defined as excessive hair that appears in male-like pattern in women. Some experts consider this condition as a result of an atypical relationship between levels of circulating androgens and sensitivity of androgen receptors in hair follicles to circulating androgens. AIMS: The aim of this research work was to evaluate the efficacy of a topical treatment for suppressing terminal hair growth of a cream containing 6.0% of the Stryphnodendron adstringens bark extract. STUDY DESIGN AND SUBJECTS: Study was randomized, double-blind and placebo-controlled. Subjects with excess of terminal hair were randomized to placebo and to the active treatment (cream with 6.0% of the extract). Evaluation was performed before and after 6 months, and subjects were photographed in each time. Clinical examination was carried out with the same physicians and in accordance with the Ferriman-Gallwey (FG) score. RESULTS: Benefits of the cream containing S. adstringens bark extract was observed in 60.98% (P < 0.001) of the subjects. FG score changed from 4 to 3 in the placebo group compared to 4-2 in the active. The cream suppressed the terminal hair growth and diminished the number of terminal hair. Subjects also described the reduction of skin hyperpigmentation, folliculitis and acne. Adverse events were not verified by physicians or patients. CONCLUSIONS: The cream with 6.0% of the S. adstringens bark extract was effective on the reduction and on the reversion of the terminal hair excess, being considered a new promissory product for such finality. PMID- 19192017 TI - Cutaneous T-cell lymphoma following treatment of rheumatoid arthritis with tumour necrosis factor-alpha blocking agents: two cases. PMID- 19192018 TI - Role of tacrolimus (FK506) 0.1% ointment WW in vitiligo in children and imperatives of combine therapy with Trioxsalen and Silymarin suspension in progressive vitiligo. PMID- 19192019 TI - Homeopathic treatment of patients with psoriasis--a prospective observational study with 2 years follow-up. AB - Design Prospective multicentre observational study. Objective To evaluate details and effects of homeopathic treatment in patients with psoriasis in usual medical care. Methods Primary care patients were evaluated over 2 years using standardized questionnaires, recording diagnoses and complaints severity, health related quality of life (QoL), medical history, consultations, all treatments, and use of other health services. Results Forty-five physicians treated 82 adults, 51.2% women, aged 41.6 +/- 12.2 (mean +/- SD) years. Patients had psoriasis for 14.7 +/- 11.9 years; 96.3% had been treated before. Initial case taking took 127 +/- 47 min. The 7.4 +/- 7.4 subsequent consultations (duration: 19.4 +/- 10.5 min) cumulated to 169.0 +/- 138.8 min. Patients received 6.0 +/- 4.9 homeopathic prescriptions. Diagnoses and complaints severity improved markedly with large effect sizes (Cohen's d= 1.02-2.09). In addition, QoL improved (SF-36 physical component score d = 0.26, mental component score d = 0.49), while conventional treatment and health service use were considerably reduced. Conclusions Under classical homeopathic treatment, patients with psoriasis improved in symptoms and QoL. PMID- 19192020 TI - Autoerythrocyte sensitization syndrome (Gardner-Diamond syndrome): review of the literature. AB - A review of the literature concerning psychogenic purpura is presented. The diagnosis is usually based on typical anamnestic data, clinical presentation (painful inflammatory skin lesions, which progressed to ecchymoses during the next 24 h) and positive diagnostic tests with intracutaneous injections of 80% solution of washed autologous erythrocytes. No pathological findings of blood coagulation parameters are usually detected. Histopathological evaluations of lesional biopsies revealed non-specific changes. Taking into account the high frequency of psychic disorders and stress dependence of skin symptoms, therapy with psychotropic drugs (according to indications) and psychotherapy are pathogenetically grounded methods of treatment in psychogenic purpura, and should be provided together with symptomatic therapy. PMID- 19192021 TI - Developing a research strategy for dietetics. PMID- 19192022 TI - The role of diet in the management of gout: a comparison of knowledge and attitudes to current evidence. AB - BACKGROUND: Evidence supports dietary modifications in the management of gout. Despite this, the degree of implementation of this evidence by nutrition professionals and rheumatologists and those affected by gout is unknown. The present study aimed to compare usual dietary practices of patients with gout to evidence for dietary management of gout and to investigate whether the knowledge and attitudes of nutrition professionals and rheumatologists reflects current evidence. METHODS: A food frequency questionnaire was used to determine usual dietary intake of patients with gout, a separate questionnaire examined gout related dietary modifications (n = 29). Online questionnaires to examine attitudes towards dietary management of gout were completed by nutrition professionals and rheumatologists. RESULTS: Proportions of participants whose reported intakes were inconsistent with current evidence for the dietary management of gout were: alcohol, n = 14 (48%); beer, n = 18 (62%); seafood, n = 29 (100%); meat, n = 7 (24%); beef/pork/lamb, n = 24 (83%); dairy products, n = 12 (41%); vitamin C supplementation, n = 29 (100%). Of the 61 rheumatologists and 231 nutrition professionals who completed the online survey, the majority considered that weight loss and decreased alcohol intake were important or very important outcomes. Proportions were lower for decreased purine intake. Thirty four (56%) rheumatologists do not refer patients with gout to dietetic services and, of those who do, the majority refer less than half. CONCLUSIONS: Overall, patients with gout in the present study were not implementing evidence for dietary management of their condition and complex dietary issues were evident. PMID- 19192023 TI - Compulsory continuing professional development: a questionnaire-based survey of the UK dietetic profession. AB - BACKGROUND: Continuing professional development (CPD) for Health Professions Council (HPC) registrants became mandatory in July 2006. Some health professions have identified external barriers to CPD participation, and other research suggests that mandatory CPD can devalue learning. The present study aimed to investigate current CPD practices of UK dietitians and to identify their attitudes towards the new mandatory requirement. METHODS: UK Dietitians were asked to participate in an online questionnaire made available via an advert placed on the British Dietetic Association's website and in an electronic newsletter. RESULTS: Of 206 respondents, 98.1% kept a CPD portfolio. Those who had undertaken the 'ABC' placement model (23.7%) were more likely to keep their portfolio up to date (P = 0.006). Only 41.3% dietitians were confident that they would currently meet the minimum CPD requirement, whereas 77.2% believed they would comply by the first audit in 2010. Some 50.5% dietitians considered their CPD time commitment insufficient due to obstacles such as workload and time constraints. A total of 96.1% respondents acknowledged the importance of undertaking CPD, with the introduction of a mandatory system appearing to provide the motivation to engage in CPD. CONCLUSIONS: UK dietitians are currently engaging in CPD. There is, however, concern regarding achievement of the compulsory requirement for the HPC 2010 audit. The findings show barriers exist to engaging in CPD activities and to maintaining a portfolio. These issues could be addressed with the introduction of protected time for CPD. PMID- 19192024 TI - Video-recorded participant behaviours: the association between food choices and observed behaviours from a web-based diet history interview. AB - BACKGROUND: Automation of dietary assessments allows participant behaviour to be captured by video observation. It also allows clinicians to identify areas that effect reporting accuracy. The present observational study describes the differences in behaviour according to the type of foods selected by participants using a dietary assessment website encompassing diet history methodology. METHODS: Eleven free-living adults with type 2 diabetes mellitus were video recorded while using a dietary assessment website in a laboratory setting. Significant observable events were identified using TRANSANA software and matched for time of occurrence with grouped food item selections from the website. The frequency and proportion of behaviour types per food group were calculated and trends of associations between food groups and observed behaviour type were determined using weighted chi-square analyses. RESULTS: Sixteen categories of observed behaviour type were constructed from 7724 significant observable events. The food groups with the highest percentage of observed behaviour types were savoury sauces (7.66%), takeaways (6.47%), sugary foods (6.27%), fats and oils (5.93%), and alcoholic beverages (5.93%), which are all socially undesirable food groups. Self-touching of the face, head movement, postural movement and movement in the chair were observed significantly more often than other observed behaviour types across all food groups. CONCLUSIONS: Behaviours related to the reporting of foods during a dietary assessment may have significant implications for dietary practice. PMID- 19192025 TI - Development and validation of a food frequency index using nutritional biomarkers in a sample of middle-aged and older adults. AB - BACKGROUND: An index of diet quality, which examines different aspects of a diet concurrently, may facilitate the identification of poor dietary habits in population sub-groups. The objectives of the present study were to develop a food frequency index (FFI) and to test its associations with nutritional biomarkers and nutrient intake. METHODS: The study comprised a cross-sectional survey among 444 adults aged 55 years and older in Vienna, Austria, and a sub-sample of 226 subjects who provided fasting blood specimen. Data from a qualitative 28-item food-frequency questionnaire were used to develop the FFI. RESULTS: FFI scores were positively correlated with plasma concentrations of beta-carotene (r = 0.26), beta-cryptoxanthin (r = 0.31), zeaxanthin (r = 0.19), lutein (r = 0.21), phylloquinone (r = 0.19), 25-hydroxyvitamin D (r = 0.20), and serum high-density lipoprotein (HDL)-cholesterol (r = 0.24) and were negatively correlated with the ratio of total : HDL-cholesterol (r = -0.23). Sub-groups with higher FFI scores had, on average, lower intakes of total fat, saturated fat, and dietary cholesterol and higher intakes of total carbohydrates, dietary fibre, and most of the examined micronutrients. CONCLUSION: The FFI as a measure of diet quality has the ability to discern population sub-groups, with reasonable validity, into low- or high-risk dietary habits. PMID- 19192026 TI - Estimation of total body water from bioelectrical impedance spectroscopy in oncology outpatients receiving radiotherapy and agreement with three prediction equations. AB - BACKGROUND: Bioelectrical impedance spectroscopy (BIS) may be more accurate in determining total body water (TBW) than bioelectrical impedance analysis (BIA). The present study compared the agreement between three TBW prediction equations developed using BIA and BIS-derived TBW in oncology outpatients. METHODS: A cross sectional, observational study was conducted in 37 outpatients receiving radiotherapy (27 males/10 females, aged 68.3 +/- 10.2 years). TBW was estimated by BIS (TBW(BIS)) and three BIA TBW prediction equations (TBW(ca-u): underweight cancer patients; TBW(ca-n): normal-weight cancer patients; and TBW(rad): patients receiving radiotherapy). Bland-Altman analyses determined agreement between methods. BIS-derived TBW using new resistivity constants was calculated. RESULTS: The mean +/- SD of TBW estimated by BIS was 39.8 +/- 8.3 L, which was significantly different from the prediction equations; TBW(rad) 35.1 +/- 7.9 L, TBW(ca-u) 33.1 +/- 7.5 L and TBW(ca-n) 32.3 +/- 7.3 L, (P < 0.001). Using new resistivity constants, TBW was 36.2 +/- 8.1 L but this still differed from the equations (P < 0.001). Bias between TBW(BIS) and that predicted by the equations was in the range 4.7-7.4 L or 1.1-3.9 L using new resistivity constants. CONCLUSIONS: TBW estimated by BIS cannot be directly compared with oncology specific BIA equations, suggesting that BIS cannot be used at the group level in outpatients receiving radiotherapy. There was a reduced bias with BIS using new resistivity constants; however, further research should determine any advantage of BIS over BIA in this population. PMID- 19192027 TI - The long-term influence of orlistat on dietary intake in obese subjects with components of metabolic syndrome. AB - BACKGROUND: Orlistat is a lipase inhibitor that reduces the intestinal absorption of fat and may enhance the effects of dietary and behavioural therapy on weight loss and maintenance. The present study examined the effect of orlistat on dietary intake, especially fat intake, during long-term weight maintenance. METHODS: Subjects comprised 44 men and women (aged 18-63 years; body mass index 37.5 +/- 4.3 kg m(-2)) included in the Scandinavian Multicenter study of Obese subjects with the metabolic syndrome, a 3-year clinical trial of orlistat or placebo following an 8-week, very low energy diet (VLED). Two months after the end of the trial when the use of orlistat was optional, 33 subjects remained in the study. A dietary interview based on a validated food frequency questionnaire was conducted before the VLED, after 1 year of treatment with orlistat or placebo and 2 months after the end of the trial. RESULTS: At 1 year, dietary intake did not differ between the orlistat and placebo group. Energy percent (E%) fat was reduced and E% carbohydrate was increased within both groups. Two months after the end of the trial, E% fat was 32.6% (SD 6.2%) in subjects that chose to take orlistat and 27.7% (SD 5.5%) in subjects not taking orlistat [between group difference -5.0% (95% confidence interval -9.2 to -0.7); P = 0.021]. CONCLUSIONS: The use of orlistat compared with placebo in a lifestyle modification programme does not appear to influence dietary intake. Subjects that chose to take orlistat after the end of the programme did not comply with dietary recommendations and this may hamper the effect of the drug. PMID- 19192028 TI - Acute effect of nut consumption on plasma total polyphenols, antioxidant capacity and lipid peroxidation. AB - BACKGROUND: Nuts have been shown to have beneficial effects on human health due to the healthy fat content; however, the effect of antioxidants (i.e. polyphenols) in nuts have not been fully investigated. The present study aimed to assess the immediate effect of a polyphenol-rich meal (75% of energy from nuts: walnuts or almonds) and a polyphenol-free meal on plasma polyphenol content, antioxidant capacity and lipid peroxidation in healthy volunteers. METHODS: Thirteen subjects participated in a randomized, crossover, intervention study. After an overnight fast, walnuts, almonds or a control meal in the form of smoothies were consumed by study subjects. Each subject participated on three occasions, 1 week apart, consuming one of the smoothies each time. Blood samples were obtained at fasting and then at intervals up to 3.5 h after consumption of the smoothies. RESULTS: There was a significant increase in plasma polyphenol concentration following both nut meals, with peak concentrations being achieved at 90 min, and with a walnut meal having a more sustained higher concentration than an almond meal. The plasma total antioxidant capacity reached its highest point at 150 min postconsumption of the nut meals, and was higher after the almond compared to walnut meal. A gradual significant (P < 0.05) reduction in the susceptibility of plasma to lipid peroxidation was observed 90 min after ingestion of the nut meals. No changes were observed following consumption of control meal. CONCLUSIONS: Consumption of both nuts increased plasma polyphenol concentrations, increased the total antioxidant capacity and reduced plasma lipid peroxidation. PMID- 19192029 TI - Evaluation of attendance and weight loss in an intensive weight management clinic compared to standard dietetic care. AB - BACKGROUND: With increasing rates of obesity, the effectiveness of weight reduction programmes come under increasing scrutiny. This nonrandomized study aimed to review the effectiveness of two weight loss clinics in terms of percent and rate of weight loss, and attendance. METHODS: Data were collected on consecutive obese patients, attending either an intensive weight management clinic (IWMC) or a general dietetic outpatient clinic. The IWMC had a structured approach with six once-a-month appointments, a signed agreement to attend, an initial screening of readiness to change and consistent advice from one dietitian. The general clinic was less structured, had more ad hoc follow up and did not guarantee one dietitian. RESULTS: Seventy percent of patients referred were female [mean (SD) age 48 (14.2) years]. Thirty-three percent (103/313) of all patients referred did not book an appointment. Of those attending with a body mass index > or = 32 kg m(-2), 55% were seen in the general and 45% in the intensive clinic, but only 19% and 53%, respectively, completed the programmes. The total amount and rate of weight loss did not differ significantly between clinics. However, analysis using the last recorded weight revealed a median weight reduction of 1.8% (interquartile range = -5.6-0) at the median rate of 0.4 kg month(-1) (-1-0) in the intensive clinic, compared to no overall weight loss in the general clinic (P < or = 0.001). CONCLUSIONS: A more structured approach and initial screening of readiness to change is likely to achieve better weight loss results and therefore will comprise a better use of dietetic time than including obese patients in general clinics. PMID- 19192030 TI - Low-glycaemic index sweetener-based beverages reduce 24-h glucose profiles in healthy adults. AB - BACKGROUND: Previous research has shown that substitution of high-glycaemic index (GI) bread for low-GI bread can favourably alter the 24-h glucose profile. Given the high beverage consumption in the UK, the present study aimed to evaluate the impact of consuming a beverage containing a low-GI, natural sweetener at mealtimes compared to a sucrose-based beverage, on the 24-h glucose profile. METHODS: In a randomized crossover design, six subjects (aged 41 +/- 16 years; body mass index = 25.8 +/- 4.1 kg m(-2)) were provided with a diet including low GI or control (sucrose) beverages on two nonconsecutive days. On each study day, subjects consumed the low-GI or control beverage at breakfast, lunch and dinner. Interstitial glucose concentrations were measured over 24 h using a continuous glucose monitoring system. RESULTS: Compared to the control beverage, the low-GI beverage significantly reduced mean glucose concentration over 24 h (P < 0.05). Similarly, 24-h and daytime incremental area under the curves for glucose were significantly lower (P = 0.001). CONCLUSIONS: The substitution of a sucrose-based beverage for a beverage containing a low-GI, natural sweetener at mealtimes is a simple dietary change that can significantly reduce 24-h glucose levels and may be beneficial for many individuals. PMID- 19192034 TI - Introduction: Euromit VII--summarizing 50 years of research on mitochondrial disease. PMID- 19192035 TI - Disorders from perturbations of nuclear-mitochondrial intergenomic cross-talk. AB - In the course of evolution, mitochondria lost their independence, and mitochondrial DNA (mtDNA) became the 'slave' of nuclear DNA, depending on numerous nucleus-encoded factors for its integrity, replication and expression. Mutations in any of these factors may alter the cross-talk between the two genomes and cause Mendelian disorders characterized by qualitative (multiple deletions) or quantitative (depletion) alterations of mtDNA, or by defective translation of mtDNA-encoded respiratory chain components. PMID- 19192036 TI - Mitochondrial medicine: entering the era of treatment. AB - Research of patients with defects in cellular energy metabolism (mitochondrial disease) has led to a better understanding of mitochondrial biology in health and disease. The obtained knowledge is of increasing importance for physicians of all medical disciplines. It assists in enabling the development of rational treatment strategies for diseases or conditions caused by mitochondrial dysfunction. The still frequently used classical interventions with vitamins or co-factors are only beneficial in some rare mitochondrial disease conditions, like coenzyme Q biosynthesis defects. For that reason alternative strategies to correct disturbed energy metabolism have to be developed. New approaches in this direction include nutrition and exercise therapies, alternative gene expression, enzyme replacement, scavenging of potentially toxic compounds and modulating cell signalling. The effect of some of these interventions has already been explored in humans whilst others are still at the level of single cell research. We review the state of the art of the development of mitochondrial treatment strategies and discuss what steps need to be taken to efficiently approach the huge burden of disease caused by dysfunctional mitochondria. PMID- 19192037 TI - Batteries not included: diagnosis and management of mitochondrial disease. AB - In 1998, Wallace et al. (Science 1988; 242: 1427-30) published evidence that the mutation m.11778G>A was responsible for causing Leber's hereditary optic neuropathy. This was the first account of a mitochondrial DNA mutation being irrefutably linked with a human disease and was swiftly followed by a report from Holt et al. (Nature 1988; 331: 717-9) identifying deletions in mitochondrial DNA as a cause for myopathy. During the subsequent 20 years there has been an exponential growth in 'mitochondrial medicine', with clinical, biochemical and genetic characterizations of a wide range of mitochondrial diseases and evidence implicating mitochondria in a host of many other clinical conditions including ageing, neurodegenerative illness and cancer. In this review we shall focus on the diagnosis and management of mitochondrial diseases that lead directly or indirectly to disruption of the process of oxidative phosphorylation. PMID- 19192038 TI - The impact of chronic multimorbidity and disability on functional decline and survival in elderly persons. A community-based, longitudinal study. AB - OBJECTIVE: We aimed to disentangle the effect of chronic multimorbidity and disability on 3-year functional decline and survival in the elderly. DESIGN: Prospective cohort study with a mean of follow-up of 2.8 years. SETTING: Swedish elderly persons from the Kungsholmen Project (1987-2000). SUBJECTS: A total of 1099 subjects, 77-100 years old, living in the community and institutions. MAIN OUTCOME MEASUREMENTS: Medical diagnoses (based on clinical examination, drug use, medical records and blood tests), and functional assessment (according to Katz Index) at baseline were investigated in relation to functional decline and death occurring during follow-up. RESULTS: At baseline, 12.1% of participants had disability, and 52.3% were affected by multimorbidity. During follow-up, 363 persons died and 85 worsened in functioning. The number of chronic conditions incrementally increased the risk of functional decline [hazard ratio (HR) increased from 1.5 in subjects with one disease to 6.2 in persons with 4+ diseases]. However, this was not the case for mortality, as the HR of death was the same for people with one disease as well as 4+ diseases (HR=2.3). Baseline disability had the highest impact on survival, independently of number of diseases [HR=8.1; 95% confidence interval (CI)=4.8-13.7 in subjects with one disease and HR=7.7; 95% CI=4.7-12.6 in those with 2+ diseases]. CONCLUSIONS: In the elderly subjects, chronic disability rather than multimorbidity emerged as the strongest negative prognostic factor for functionality and survival. PMID- 19192039 TI - Is endometroid cancer of the ovaries a late manifestation of Alstrom syndrome? PMID- 19192040 TI - Skin signs in acute pancreatitis: frequency and implications for prognosis. PMID- 19192041 TI - Closing the clinical-imaging gap in multiple sclerosis? Imaging iron deposition in deep gray matter. PMID- 19192042 TI - Deep gray matter involvement on brain MRI scans is associated with clinical progression in multiple sclerosis. AB - BACKGROUND: Conventional brain MRI lesion measures have unreliable associations with clinical progression in multiple sclerosis (MS). Gray matter imaging may improve clinical-MRI correlations. METHODS: We tested if gray matter MRI measures and conventional measures of lesions/atrophy predicted clinical progression in a 4-year longitudinal study of 97 patients with MS. Baseline and follow-up brain MRI were analyzed for basal ganglia and thalamic normalized T2 signal intensity, whole brain T2-hyperintense lesion volume, and whole brain atrophy. Logistic regression tested the ability of baseline or on-study change in MRI to predict disability progression, as reported by area under the receiver operator characteristics curve (AUC). RESULTS: Lower caudate T2-intensity at baseline (P= .04; AUC = .69) and on-study decreasing T2-intensity in the putamen (P= .03; AUC = .70) and thalamus (P= .01; AUC = .71) were the MRI variables associated with clinical progression when regression modeling was adjusted for length of follow up interval, baseline EDSS, disease duration, age, and sex. CONCLUSIONS: Gray matter T2-hypointensity, suggestive of excessive iron deposition is associated with worsening disability in patients with MS. Gray matter MRI assessment may be able to capture neurodegenerative aspects of the disease, with more clinical relevance than derived from conventional MRI measures. J Neuroimaging 2009;19:3 8. PMID- 19192043 TI - Self-referral of imaging does not imply overutilization. AB - For several years, some sectors of the specialty of Radiology have complained about the practice of self-referral, where a nonradiologist physician provides and interprets an imaging procedure. It is argued that such practice leads to increased costs since a physician will overutilize technology because of financial incentives. Here we review the literature. The most extensive analysis to date is at odds with the conclusions drawn in the older literature in that it provides little, if any, evidence for overutilization. We performed our own investigation using a poll study and found no suggestion of overutilization in 33 self-referring neurologists when compared with 900 neurologists who referred imaging to radiologists. The main period of growth in demand for imaging was between 1999 and 2002. Since 2002 there has been a steep decline in the rate of growth, so that it is possible to predict roughly zero growth in MRI utilization by 2009 without any intervention. It is shown why the rise in demand for imaging studies cannot be explained by self-referral, and it is argued that the sudden expansion of demand 8 years ago was caused by simultaneous technological improvements in the 3 major imaging modalities. Finally, it is shown how self referral may actually reduce costs by facilitating the transfer of care from the hospital and ER to the office. PMID- 19192046 TI - Altered binding of MYF-5 to FOXE1 promoter in non-syndromic and CHARGE-associated cleft palate. AB - BACKGROUND: Three different homozygous loss-of-function mutations of the Forkhead box E1 (FOXE1) gene have been associated with syndromic cleft palate. Here, we screened the entire promoter region to identify the variations in significant consensus motifs affecting FOXE1 transcription. METHOD: Genomic DNAs of 35 cleft palate patients, 10 of whom with CHARGE association, 80 unrelated healthy people and 80 unaffected first-degree relatives were analysed by automatic sequencing. The Transcription Element Search System program was employed to identify transcription factor binding sites. The protein-DNA complexes were observed using DNA band-shift assays and oligonucleotide competition analyses. Real-time PCR was used to estimate FOXE1 expression at mRNA level. RESULTS: In 11 non-syndromic cleft palate patients, a novel non-coding polymorphism (C-->G) in the 5' untranslated region of FOXE1 was found. The variation fell into a putative consensus sequence for the transcription factor MYF-5 and completely impaired the ability of MYF-5 to bind to its motif, as shown by EMSA experiments. As a consequence, a significantly reduced FOXE1 mRNA expression was observed. CONCLUSIONS: In 45% of non-syndromic cleft palate patients, a novel homozygous polymorphism that prevented the binding of MYF-5 to FOXE1 promoter and affected the FOXE1 expression was found. As recent data show the role of MYF-5 in the muscle-dependent craniofacial skeletal development and in the fusion of primary palate and secondary palate, the results reported here strongly suggest a more significant involvement of this factor in the cleft palate onset. PMID- 19192047 TI - A case-control study on etiological factors involved in patients with burning mouth syndrome. AB - OBJECTIVE: To evaluate and analyze the risk factors for burning mouth syndrome (BMS). METHODS: Eighty-seven consecutive patients with BMS and a randomly selected control group (n = 82) were comprehensively investigated with a self designed questionnaire, Self-rating Anxiety Scale (SAS) and Self-rating Depression Scale (SDS). A complete blood count and serum sex hormone were also examined in patients with BMS and control subjects. All the data obtained were transferred to a data bank and analyzed statistically in SPSS 11.5 for windows. RESULTS: No statistical difference between the BMS group and the control group was found in blood analyses including white blood cell count, red blood cell count, hemoglobin and platelet count. Among the menopausal or postmenopausal women with BMS, the follicle stimulating hormone (FSH) level was significantly higher, but the estradiol level was significantly lower. The BMS group reported adverse life events more frequently than the control group. Patients with BMS significantly exhibited symptoms of somatization, and both the scores of anxiety, depression in patients with BMS were higher than those of the control group (P < 0.05). A regression equation which included six variables had been established by using logistic regression analysis, indicating that the habit of tongue thrusting, lip sucking, periodontitis, smoking, outcome of recent medication, depression were the principal risk factors, among which tongue thrusting was the most significant. CONCLUSION: Our study indicated that BMS may be of psychological origin, and the measures such as refraining from oral parafunctional activities, removing local irritating factors, stopping smoking, good mental health status could help in the prevention of BMS. PMID- 19192048 TI - Calcitonin gene-related peptide levels in saliva of patients with burning mouth syndrome. AB - Burning mouth syndrome (BMS) is an intraoral burning sensation for which no medical or dental cause can be found. Recent studies suggest that primary neuropathic dysfunction might be involved in the pathogenesis of BMS. Calcitonin gene-related peptide (CGRP) plays an important role in the development of pain and serves as a biological marker of trigeminovascular activation. The aim of this study was to determine the levels of CGRP in the saliva of BMS patients and estimate the trigeminovascular activation in BMS. CGRP levels were measured, by RIA method in 78 BMS patients and 16 healthy subjects. The levels of CGRP were non-significantly decreased in BMS patients in comparison to healthy subjects. These results suggest that trigeminal nerve degeneration may be the underlying cause of BMS. PMID- 19192050 TI - Immunohistochemical analyses of survivin and heat shock protein 90 expression in patients with oral lichen planus. AB - BACKGROUND: Survivin is an inhibitor of apoptosis protein. Heat shock protein 90 (Hsp90) is a molecular chaperone that is involved in protein folding and maturation. Alterations in the expression of survivin and Hsp90 have been reported in several chronic inflammatory diseases. No study has evaluated the expression of survivin and Hsp90 in the same lesion of oral lichen planus (OLP). The objective of this study was to investigate the expression of survivin and Hsp90 in biopsy specimens of patients with OLP and controls. METHOD: Formalin fixed and paraffin-embedded biopsy specimens from 29 patients with OLP and 29 healthy subjects were used in this study. Immunohistochemical methods were used to detect the expression of survivin and Hsp90 in the oral epithelia of these two groups. The distributions of positively stained epithelial cells were evaluated with a semiquantitative method, using an immunostaining score system. RESULTS: Expression of survivin and Hsp90 in oral epithelia of patients with OLP was significantly different in comparison with that in controls (P = 0.001 and P < 0.001 respectively). These differences were in the direction towards reduction in the expression of survivin and Hsp90 in the oral epithelia of patients with OLP. CONCLUSION: Our results provided additional evidence that alterations in the expression of survivin and Hsp90 in oral epithelia might be associated with pathological stress, such as chronic inflammation, in patients with OLP. PMID- 19192049 TI - Activation of innate immune responses through Toll-like receptor 3 causes a rapid loss of salivary gland function. AB - BACKGROUND: Recent studies have demonstrated the expression of Toll-like receptor 3 (TLR3) in salivary glands and epithelial cell lines derived from Sjogren's syndrome (SS) patients. As viral infections are considered to be a trigger for SS, in this study we investigated whether in vivo engagement of TLR3 affects salivary gland function. METHODS: Female New Zealand Black/WF1 mice were repeatedly injected with polyinosinic:polycytidylic acid [poly(I:C)]. TLR3 expression within submandibular glands was studied using immunohistochemistry. RNA levels of inflammatory cytokines in the submandibular glands were determined by real time polymerase chain reaction. Pilocarpine induced saliva volume was used as an index of glandular function. RESULTS: Immunohistochemical analysis of submandibular glands showed TLR3 expression in epithelium of serous and mucous acini, granular convoluted tubules, and ducts. Poly(I:C) treatment rapidly up regulated the mRNA levels of type I interferon (IFN) and inflammatory cytokines in the submandibular glands. One week after treatment, the saliva volumes in poly(I:C) treated mice were significantly reduced in comparison with the phosphate-buffered saline (PBS) treated mice. Hematoxylin and eosin staining showed that salivary gland histology was normal and lymphocytic foci were not detected. Glandular function recovered after poly(I:C) treatment was stopped. CONCLUSIONS: Our results demonstrate that engagement of TLR3 within the salivary glands results in a rapid loss of glandular function. This phenomenon is associated with the production of type I IFN and inflammatory cytokines in the salivary glands. Restoration of glandular function suggests that for viral etiology of SS, a chronic infection of salivary glands might be necessary. PMID- 19192051 TI - Polymorphic drug metabolizing CYP-enzymes--a pathogenic factor in oral lichen planus? AB - BACKGROUND: Oral lichen planus (OLP) is a chronic mucosal disease with a characteristic clinical phenotype. Environmental exposures, e.g. drugs have been associated with the pathogenesis. OBJECTIVES: To test the hypothesis that some OLP lesions have a pharmacological pathogenesis related to polymorphisms of the cytochrome P450 enzymes (CYPs) resulting in poor or intermediate CYP metabolism. METHODS: One hundred and twenty patients with OLP and 180 gender-matched controls without OLP were genotyped for CYP2C9, CYP2C19, and CYP2D6 alleles with absent or reduced function. RESULTS: The prevalence of poor or intermediate metabolizers was not higher among the OLPs as compared with the controls; however, there were higher numbers of variant CYP2D6 genotypes among the OLP females (P < 0.05). There were no differences between the groups with regard to intake of drugs metabolized by polymorphic CYPs or drug or herbal products inhibiting CYPs. The prevalence of CYP2D6*4 alleles among the OLPs was higher [28%; 95% confidence interval (CI) 20-36%] than previously reported among Danes (19%; 95% CI 17-22%). Fifty per cent of the OLPs had a CYP2D6*4 genotype as compared with 30% in the background population (P = 0.0001). The CYP2D6*4 protein has sequence homology with human herpes simplex virus type 1 (HSV1) and Candida albicans, which may result in molecular mimicry. CONCLUSION: It was not possible to substantiate a pharmacological pathogenesis of OLP based on poor or intermediate CYP metabolism. However, molecular mimicry between CYP2D6, in particular CYP2D6*4, and common oral pathogens may be involved in the pathogenesis of OLP. PMID- 19192052 TI - Molecular markers in the surgical margin of oral carcinomas. AB - BACKGROUND: Local or regional lymph node recurrence is the most common pattern of treatment failure in oral squamous cell carcinoma (SCC). The local recurrence rate is 30% even when the surgical resection margin is diagnosed as tumour free. Accumulation of genetic changes in histologically normal epithelium in the surgical resection margin may explain the local recurrence rate. The purpose of this study is to investigate the presence of senescence markers, which may represent early malignant changes in the margin that in routine pathological evaluations are classified as histologically normal. METHODS: Formalin-fixed, paraffin-embedded surgical specimens from 16 consecutive patients with oral SCC and a clear surgical margin were obtained. The margin was analysed by immunohistochemistry for p53, p16, Chk2, Laminin-5 and glycosylated oncofetal fibronectin. RESULTS: Two patterns of p53 expression were found in the histologically normal epithelium in the surgical resection margin. One was characterized by no protein expression in the majority of cells, except for small clusters of basal and parabasal cells with nuclear staining. The other was characterized by p53 expression in the nuclei of most basal cells. The expression of p16 was confined to small groups of cells in the basal cell layer whereas Chk2 was only seen in one case. Upregulation of the stromal proteins, Laminin-5 or glycosylated oncofetal fibronectin, was only seen at regions of invasion. CONCLUSION: Small groups of cells expressing p53 and p16 were found in the surgical resection margin that appeared to be histologically normal and may represent early malignant changes. PMID- 19192053 TI - Areca nut extract-treated gingival fibroblasts modulate the invasiveness of polymorphonuclear leukocytes via the production of MMP-2. AB - BACKGROUND: Areca nut chewing is associated with an increase in the incidence of oral neoplastic or inflammatory diseases. Aberrations in matrix metalloprotease (MMP) expression are associated with the pathogenesis of oral diseases. This study investigated the potential effects of areca nut extract (ANE) on human gingival fibroblasts and the consequential impacts on inflammatory pathogenesis. METHODS: Analyses of senescence marker, cell viability, changes of the cell cycle, and cell granularity in gingival fibroblasts together with an assessment of the invasiveness of polymorphonuclear (PMN) leukocytes after treatment with the supernatant of ANE-treated gingival fibroblasts were performed to characterize the phenotypic impacts. Western blotting and gelatin zymography were used to assay the expression and activity of MMP-2. RESULTS: Chronic subtoxic (<10 microg/ml) ANE treatment resulted in premature growth arrest, appearance of senescence-associated beta-galactosidase activity and various other senescence associated phenotypes in gingival fibroblasts. Gingival fibroblasts established from older individuals had a higher propensity to become ANE-induced senescent gingival fibroblasts. An activation of MMP-2 was identified in senescent cells. PMN leukocytes treated with the supernatant of ANE-induced senescent cells exhibited a significant increase in invasiveness, which was abrogated by both a MMP-2 blocker and a MMP-2 nullifying antibody. CONCLUSIONS: This study provides evidence whereby MMP-2 secreted from ANE-induced senescent gingival fibroblasts would facilitate the invasiveness of PMN leukocytes, which could be associated with the oral inflammatory process in areca chewers. PMID- 19192054 TI - Methylation of P16, P21, P27, RB1 and P53 genes in odontogenic keratocysts. AB - BACKGROUND: Odontogenic keratocyst (OKC) is a benign neoplasm with an aggressive clinical behavior and a high recurrence rate. Although epigenetic alterations have been reported in different tumors, these events were not investigated in OKC yet. The aim of this study was to investigate the presence of methylation in P16, P21, P27, P53 and RB1 genes in OKC tumors. METHODS: Methylation-specific polymerase chain reaction (MSP) was used to evaluate the presence of methylation in 10 samples of OKCs, 10 samples of dental follicles and six samples of normal mucosa. RESULTS: The methylation status of the P16 gene was similar among the three groups. In P21 gene, 30% of OKCs were methylated while no methylation could be detected in the other groups. High frequency of P27 methylation (90%) was observed in dental follicles, however, some OKC lesions (10%) and normal mucosa samples (33%) were also methylated. Concerning the RB1 gene, positive results were detected only in dental follicles (40%). No positive result was observed considering P53 gene. CONCLUSIONS: Our data show methylation of the promoter of P21 gene in OKCs. In addition, methylation of the P27 and RB1 genes are commonly found in dental follicles. Further studies are necessary to determine the functional relevance of these alterations. PMID- 19192055 TI - Expression of p16 in oral cancer and premalignant lesions. AB - BACKGROUND: Expression of p16 has been proposed as a marker for malignant transformation. This study aimed to evaluate p16 expression in oral squamous cell carcinoma (OSCC) and premalignant lesions including oral leukoplakia (OL) with and without dysplasia. METHODS: Expression of p16 was investigated in 56 samples including OSCC, OL with and without dysplasia, and normal oral mucosa. Expression of p16 was identified by immunohistochemistry, using the CINtecTM p16INK4a Histology Kit. Both nuclear and/or cytoplasmic staining of the keratinocytes were considered to be positive and the percentage of positive cells was calculated. RESULTS: Expression of p16 was detected in 3/16 (18.75%) cases of OSCC, in 4/15 (26.7%) cases of OL without dysplasia, and in none of OL with dysplasia and normal mucosa. No significant differences in p16 expression prevalence were found among OSCC, OL with and without dysplasia and normal mucosa. The percentages of positive cells in OSCC and OL without dysplasia were 0.89 and 0.17, respectively. No significant difference in the percentage of positive keratinocytes was found. CONCLUSION: As a marker, p16 is not reliable for oral mucosal dysplasia and malignant transformation. PMID- 19192056 TI - An investigation of the role of oral epithelial cells and Langerhans cells as possible HIV viral reservoirs. AB - BACKGROUND: The role of the oral mucosa as a target of human immunodeficiency virus (HIV-1) infection and persistence is unclear. HIV-1 has been reported in oral epithelial cells, but this has not been confirmed. Cellular reservoirs may impede antiretroviral therapies and should be identified. This study was performed to determine the presence of HIV-1 in oral epithelial and Langerhans cells (LCs) of HIV-1-positive antiretroviral naive patients. Non-invasive brush biopsy technique for future in vivo HIV research was also evaluated. METHODS: Oral mucosal cells were harvested from the buccal mucosae, dorsal tongue and the gingiva of the mandibular teeth of 35 HIV-1-positive patients using a Cytobrush Plus cell collector. Epithelial cells were purified from the samples by flow cytometric cell sorting using cytokeratin stains after which the epithelial cell samples were further purified and divided into superficial and deep epithelial cells by laser microdissection on Pap stained cytospin smears. LCs were picked up individually by laser microdissection from CD1a stained cytospin smears. Purified epithelial and LC samples were tested for the presence of HIV-1 DNA by polymerase chain reaction analysis. RESULTS: Ten of the patients had HIV-1 DNA in one or more of the sampled anatomical locations. No HIV-1 DNA could be demonstrated in any of the purified superficial or deep epithelial or LC samples. CONCLUSIONS: HIV-DNA can be found using non-invasive oral brush biopsies and should be investigated further as an experimental model for in vivo oral HIV research. Better ways to purify the different cell types should be investigated. PMID- 19192057 TI - Oral ulcers in HIV-positive Peruvian patients: an immunohistochemical and in situ hybridization study. AB - BACKGROUND: This study describes the histopathological, immunohistochemical (IHC) and in situ hybridization (ISH) data of 25 cases of oral ulcers in HIV-positive patients, with clinical and microscopical features similar to ulcers not otherwise specified (NOS)/necrotizing ulcerative stomatitis (NUS). METHODS: Sex, age and clinical history were obtained from the clinical records. Histological analysis for H&E, Gomori-Grocott and Ziehl-Neelsen stains, IHC analysis to detect infectious agents and to characterize inflammatory cellular infiltrate, and ISH for cytomegalovirus (CMV) and EBER1/2 were performed. RESULTS: Twenty-one patients were men and four were women (mean age of 34.6 years). The tongue was preferentially affected. Microscopically, the lesions showed extensive necrosis, leukocytoclasia, vasculitis with luminal fibrin clots and an intense inflammatory cellular infiltrate predominated by CD68(+) atypical large cells, normal-sized and crescent-shaped nuclei macrophages, interspersed by CD8(+) T lymphocytes. Mast cells were also observed in all samples studied. CD4(+) T lymphocytes, CD20(+) B lymphocytes and VS38c(+) plasma cells were practically absent. CMV and EBER1/2 were identified in scarce cells of 3 and 16 of 25 cases respectively. CONCLUSION: These results show that CD68(+) macrophages, followed by CD8(+) T lymphocytes, were the predominant inflammatory cells, indicating they are relevant to the pathogenesis of the ulcers, possibly reflecting an abnormal immune response in the oral mucosa. The clinicopathological and immunoprofile features of the present cases are similar to NOS ulcers/NUS in HIV-positive patients. PMID- 19192058 TI - Oral manifestations of HIV infection in adult patients from the province of Sancti Spiritus, Cuba. AB - BACKGROUND: Studies on the prevalence of HIV-related oral lesions (HIV-OL) have shown great variations among different countries. The aim of this study was to describe the prevalence of HIV-OL in adults infected with HIV in the province of Sancti Spiritus, Cuba, and to determine the factors associated with the presence of HIV-OL. METHODS: A cross-sectional observational study was performed between November 2006 and August 2007 at the Hospital General Universitario 'Camilo Cienfuegos', Sancti Spiritus. One hundred and fifty-four HIV-infected patients were included. Patients were examined and interviewed by a periodontal specialist. Diagnosis of HIV-OL was based on clinical criteria. Demographical, clinical and laboratory data were obtained. Independent association of each factor with HIV-OL was assessed by logistic regression modelling. RESULTS: The prevalence of HIV-OL was 40.9%. The commonest manifestation was oral hairy leucoplakia (n = 19; 12.3%); oral candidiasis (n = 17; 11%); herpes simplex virus infection (n = 11; 7.4%); and aphthous ulcer (n = 9; 5.8%). Principal factors associated with the presence of HIV-OL were CD4(+) lymphocytes <500 cells/mm(3) (OR: 2.06; 95% CI: 1.019-4.195) and smoking (OR: 2.03 CI: 1.037-3.982). CONCLUSION: This study described the prevalence of HIV-OL in 154 HIV-infected patients which represent about 80% of those known to be infected in the province of Sancti Spiritus. The prevalence of HIV-OL was lower than those reported from developing countries. Oral hairy leucoplakia and oral candidiasis were the most prevalent HIV-OL. Smoking and CD4(+) cells count <500 cells/mm(3) were the two factors independently associated with the presence of HIV-OL. PMID- 19192059 TI - Granular cell tumor of the oral cavity: updated immunohistochemical profile. AB - BACKGROUND: Granular cell tumor (GCT) is a benign lesion that occurs at different body sites with preponderance to the oral cavity. It is generally believed to be of schwann cell/neural cell origin. We used a large panel of both traditional and recently developed antibodies in an attempt to trace the origin of GCTs on the basis of their immunoprofile. METHODS: The patients' demographic data and the cytological and architectural features of the lesions were analyzed in a large series of oral GCTs (n = 68). Forty-two lesions were also submitted to a panel of immunohistochemical stains with antibodies against S-100, CD-68 (KP-1 and PG-M1), vimentin, calretinin, NKI/C3, PGP9.5, p75/NGFR and inhibin-alpha. RESULTS: The tongue was the most common location of oral GCTs (81%). The granular cells demonstrated a wide array of cytological features in terms of cell shape and position of the nucleus. In addition, the lesions showed different architectural patterns, including 'infiltration' with satellite nodules. Interestingly, no recurrences were reported, even in lesions that were not completely excised. Granular cells were usually found to be strongly and diffusely positive for p75, vimentin, calretinin and NKI/C3, inhibin-alpha, PGP9.5, and S-100. CONCLUSIONS: Immunoreactivity of the granular cells to a broad panel of antibodies that characterize different tissues does not confirm any particular cell type for the histogenetic origin of GCTs. Furthermore, GCTs could be regarded as lesions that reflect a local metabolic or reactive change rather than a true neoplasm. PMID- 19192060 TI - A reappraisal of diagnostic criteria for mucous membrane pemphigoid. PMID- 19192062 TI - Inflammatory Neuropathy Consortium: Report of a meeting held on 4-6 July 2008, Batiment Babinski, Hopital de la Salpetriere, Paris, France. AB - The Inflammatory Neuropathy Consortium held its first scientific meeting in Paris on 4-6 July 2008. The consortium is a special interest group of the Peripheral Nerve Society. The meeting included 10 plenary lectures and over 60 abstracts. A meeting summary is provided. PMID- 19192063 TI - What's new in chronic inflammatory demyelinating polyradiculoneuropathy in 2007 2008? AB - Chronic inflammatory demyelinating polyradiculoneuropathy (CIDP)-related research has made progress in the field of pathogenesis, genetics, and treatment. The number of circulating CD4(+) CD25(+) T-regulatory cells was shown to be reduced in CIDP patients. Increased frequency of genotype GA13-16 of the SH2D2A gene encoding for a T-cell-specific adapter protein in CIDP patients may result in a defective control and elimination of autoreactive T cells. IVIg treatment has been shown to increase numbers and function of peripheral CD4(+) CD25(+) T regulatory cell in a mouse model. These findings shed new light on the understanding of why peripheral tolerance is breached in CIDP patients and why the disease becomes chronic and adds another possible mechanism of action of intravenous immunoglobulin to the already long list. Long-term effectiveness of IVIg has now been proven. Subcutaneous immunoglobulin could be an alternative for IVIg, but this has to be explored further in well-designed trials. Autologous stem cell transplantation has been tried in refractory patients, but larger trials are necessary to assess safety and effect of this treatment. PMID- 19192064 TI - What's new in multifocal motor neuropathy in 2007-2008? AB - Several articles on multifocal motor neuropathy (MMN) have appeared in 2007 and 2008, broadening the boundaries of this neuropathy and improving its diagnosis. A number of retrospective studies confirmed the prolonged efficacy of high-dose intravenous immunoglobulins (IVIg) in this neuropathy. The efficacy of other immune therapies was not however, confirmed in other studies in MMN including a randomized controlled trial on mycophenolate mofetil as adjunctive therapy to IVIg. These negative results and the lack of new other studies on the cause of MMN should lead to some reconsideration on its pathogenesis. PMID- 19192065 TI - What's new in paraproteinemic demyelinating neuropathy in 2007-2008? AB - Research and development in the area of paraproteinemic demyelinating peripheral neuropathy is slower than for some of the other inflammatory neuropathy syndromes. However, 2007-2008 has produced developments in the understanding of the pathogenesis of the neuropathy and the paraprotein, a potential new animal model, easier diagnostics, and two randomized controlled trials with the results of others awaited. This short review summarizes these publications. PMID- 19192066 TI - Role of a pre-existing neuropathy on the course of bortezomib-induced peripheral neurotoxicity. AB - We investigated a series of bortezomib-treated patients and correlated the course of bortezomib-induced peripheral neurotoxicity with the presence or absence of peripheral neuropathy at baseline. Forty-eight patients were examined with the total neuropathy score reduced version (TNSr), visual analogue score (VAS) for pain, and nerve conduction studies at baseline and after two and four cycles of chemotherapy. Twenty-three patients had a baseline TNSr = 0-2, and 25 patients had a baseline TNSr >2 (median = 6, range 3-13). The course of bortezomib-induced peripheral neurotoxicity was generally more severe in those patients with the highest baseline TNSr. However, among those subjects with a normal baseline TNSr, two patients developed a clinically relevant peripheral neuropathy with a marked increase in TNSr as early as after two cycles of bortezomib treatment (TNSr = 10 and 15, respectively), while after four cycles, three other patients with normal baseline TNSr had a TNSr of 11, 12, and 13. VAS reporting confirmed that painful neuropathy is frequent after bortezomib administration. Our results indicate that the course of bortezomib-induced peripheral neurotoxicity can be severe in subjects with normal neurological examination at baseline, and therefore, careful monitoring during treatment is suggested in these patients. PMID- 19192067 TI - Characteristics of bortezomib- and thalidomide-induced peripheral neuropathy. AB - Dose-limiting peripheral neuropathy (PN) is frequently reported with the use of thalidomide and bortezomib, novel proteasome inhibitors. While these two agents have significant activity in multiple myeloma (MM), the combination and the associated PN have not been fully examined in untreated patients. The objective of this study was to report the baseline prevalence and occurrence of PN in newly diagnosed MM patients treated with bortezomib and thalidomide. Twenty-seven patients (11 men and 16 women) with previously untreated MM were prospectively monitored for PN. Total neuropathy score reduced (TNSr) was calculated at baseline and after every two cycles of bortezomib treatment. The median cumulative dose of bortezomib was 35.6 mg/m(2) (median 8 cycles) and of thalidomide was 16.8 g. Only three subjects showed mild PN at baseline (whole group median TNSr 0). At the end of treatment, PN developed in 26 patients (median TNSr 8). PN was of mild to moderate severity (TNSr grade 1 = 11, grade 2 = 10, grade 3 = 5, and grade 4 = 0). Nerve conduction studies showed axonal physiology in all except three subjects in whom demyelinating physiology was noted. The median TNSr was 17 in the demyelinating group and 9 in the axonal group. There was no significant correlation of TNSr with cumulative bortezomib or thalidomide dose. At follow-up, 80% of patients had become asymptomatic after discontinuation of the chemotherapy. We conclude that bortezomib and thalidomide combination chemotherapy induces a reversible length-dependent sensory>motor, predominantly axonal, large-fiber>small-fiber polyneuropathy. In a subset, a more severe demyelinating polyneuropathy may develop. PMID- 19192068 TI - Short- and intermediate-term efficacy of buprenorphine TDS in chronic painful neuropathies. AB - Buprenorphine is a potent opioid available as a transdermal delivery system (TDS) formulation. This open-label study investigated its safety, tolerability, and efficacy in 30 patients with chronic painful neuropathy. Subjects with visual analogue scale (VAS) score > or = 5 under stable analgesic treatment were entered. The starting dosage of 35 microg/h was increased up to 70.0 microg/h in case of unsatisfactory pain control as assessed by fortnightly visits. The primary endpoint was the number of patients achieving at least 30% pain relief at day 42 visit. Treatment was safe over the study period. Nine patients dropped out for side effects, mostly nausea and daily sleepiness. Buprenorphine TDS was well tolerated in 21 patients. Thirteen patients achieved > 30% of pain relief at day 42 visit. Five patients needed to increase the dosage to 52.5 microg/h. Eight patients did not meet the primary outcome, but none allowed increasing the dosage to 70 microg/h, and four patients withdrew consent to continue the study before day 42 visit because of a 'fear to become addicted,' although 40% had obtained VAS reduction. In our study, which needs to be confirmed by a controlled trial, buprenorphine TDS induced clinically meaningful pain relief in about 40% of patients with chronic painful neuropathy, suggesting its use as a third-line treatment. PMID- 19192069 TI - Inhibition of inducible nitric oxide synthase reduces an acute peripheral motor neuropathy produced by dermal burn injury in mice. AB - The systemic inflammatory response produced by a full-thickness dermal burn injury is associated with a peripheral motor neuropathy. We previously reported that a 20% body surface area (BSA) full-thickness dermal burn in C57BL6 mice produced structural and functional deficits in motor axons at a distance from the burn site. The etiology of the neuropathy, however, is not well characterized. Burn injury leads to an increase in production of a number of proinflammatory mediators, including nitric oxide (NO). We tested the hypothesis that dermal burn induced motor neuropathy is mediated by increased production of NO. NO synthase (NOS) activity was inhibited following a 20% BSA full-thickness burn by injection of non-specific NOS inhibitor, nitro-L-arginine methyl ester or inducible NOS (iNOS) inhibitors, L-N6-(1-iminoethyl) lysine, and aminoguanidine. NOS inhibitors also prevented the reduction in ventral roots mean axon caliber and the decrease in a motor nerve conduction velocity (MCV) following burn. iNOS knockout mice prevented MCV decrease in the first 3 days post-burn, but iNOS knockout MCV was significantly reduced at 7-14 days post-burn. These results suggest that an increase in NO production generated by systemic inflammatory response pathways after burn injury contributes to the development of structural and functional deficits in peripheral motor axons. PMID- 19192070 TI - Diabetes mellitus exacerbates motor and sensory impairment in CMT1A. AB - Charcot-Marie-Tooth disease type 1A (CMT1A) is caused by a duplication of PMP22 on chromosome 17 and is the most commonly inherited demyelinating neuropathy. Diabetes frequently causes predominantly sensory neuropathy. Whether diabetes exacerbates CMT1A is unknown. We identified 10 patients with CMT1A and diabetes and compared their impairment with 48 age-matched control patients with CMT1A alone. Comparisons were made with the Charcot-Marie-Tooth disease (CMT) neuropathy score (CMTNS) and by electrophysiology. The CMTNS was significantly higher in patients with diabetes (20.25 +/- 2.35) compared with controls (15.19 +/- 0.69; p = 0.01). Values were particularly higher for motor signs and symptoms. Seven of the 10 diabetic patients had CMTNS >20 (severe CMT), while only 7 of the 48 age-matched controls had scores >20. There was a trend for CMT1A patients with diabetes to have low compound muscle action potentials and sensory nerve action potentials, although nerve conduction velocities were not slower in diabetic patients compared with controls. Diabetes was associated with more severe motor and sensory impairment in patients with CMT1A. PMID- 19192071 TI - Pain accompanies pure motor Guillain-Barre syndrome. PMID- 19192072 TI - Two patients with CIDP deteriorated following plasma exchange. PMID- 19192073 TI - CMT1A duplication: refining the minimal adult phenotype. PMID- 19192074 TI - Looking again, and harder, for a link between low self-esteem and aggression. AB - Recent field studies have revived the hypothesis that low self-esteem causes aggression. Accordingly, we reanalyzed the data from a previous experiment and conducted a new experiment to study direct physical aggression in the form of blasting a fellow participant with aversive noise. We also conducted a field study using a measure of indirect aggression in the form of a consequential negative evaluation. High narcissists were more aggressive than others but only when provoked by insult or humiliation and only toward the source of criticism. The combination of high self-esteem and high narcissism produced the highest levels of aggression. These results support the view of aggression as stemming from threatened egotism and are inconsistent with the hypothesis that low self esteem causes either direct or indirect aggression. PMID- 19192075 TI - Implicit but not explicit affectivity predicts circadian and reactive cortisol: using the implicit positive and negative affect test. AB - Self-report measures assess mental processes or representations that are consciously accessible. In contrast, implicit measures assess automatic processes that often operate outside awareness. Whereas self-report measures have often failed to show expected relationships with endocrine stress responses, little effort has been made to relate implicit measures to endocrine processes. The present work examines whether implicit affectivity as assessed by the Implicit Positive and Negative Affect Test (IPANAT) predicts cortisol regulation. In Study 1, implicit low positive affectivity, but not negative affectivity, significantly predicted circadian cortisol release. In Study 2, implicit negative affectivity, but not positive affectivity, significantly predicted the cortisol response to acute stress. By contrast, cortisol regulation was not predicted by self-reported affectivity. The findings support the use of implicit affectivity measures in studying individual differences in endocrine stress responses and point to a differential role of positive and negative affectivity in baseline versus stress contingent cortisol release, respectively. PMID- 19192076 TI - The factor structure of Chinese personality terms. AB - From the Contemporary Chinese Dictionary, 3,159 personality descriptors were selected and then ranked by the frequency of use. Among those, the top 413 terms with the highest frequency were administered to two independent large samples in China for self-ratings and peer ratings to explore the emic Chinese personality structure as well as to test the universality of other models. One- and two factor structures found in previous studies of other languages were well replicated. Previous structures with more than two factors were not well replicated, but six- and seven-factor models were at least as well supported as the Big Five. Emic analysis indicated that a seven-factor structure was the most informative structure relatively salient across subsamples of self-ratings and peer ratings, across original and ipsatized data, and across differences in variable selections. These factors can be called Extraversion, Conscientiousness/Diligence, Unselfishness, Negative Valence, Emotional Volatility, Intellect/Positive Valence, and Dependency/Fragility. PMID- 19192077 TI - Links between anxiety and allergies: psychobiological reality or possible methodological bias? AB - The objective of the study was to examine the link between anxiety and allergies to establish whether it reflects a psychobiological reality or a possible methodological bias. A cohort of 1,037 children enrolled in the study. Anxiety disorders were assessed between 11 and 21 years. Anxious personality was assessed at 18 years. Allergies were examined at 21 years by (a) self reports, (b) skin pricks, and (c) serum total immunoglobulin E (IgE). Self-reported allergies were predicted by recurrent anxiety disorders (OR [95% CI]=1.56 [1.06-2.30], p=.023) and self-reports of anxious personality (OR [95% CI]=1.67 [1.17-2.37], p=.004): Objectively verified allergies were not. These results suggest that the link between anxiety and allergies may reflect a methodological artifact rather than a psychobiological reality. PMID- 19192080 TI - Individual quality of life of people with severe mental disorders. AB - People with mental disorders have been found to suffer from impaired quality of life (QoL). Therefore, the assessment of QoL has become important in psychiatric research. This explorative study was carried out in acute psychiatric wards. Thirty-five patients diagnosed with schizophrenia and related psychosis were interviewed. QoL was rated by the Schedule for Evaluation of Individual Quality of Life which is a respondent-generated QoL measure using semi-structured interview technique. Patients named five areas of life important to them and then rated their current status and placed relative weight on each QoL area. The data were analysed with qualitative content analysis and descriptive statistics. The most frequently named areas for QoL were health, family, leisure activities, work/study and social relationships, which represented 72% of all QoL areas named. Patients' average satisfaction with these QoL areas ranged 49.0-69.1 (scale 0-100). The mean global QoL score was 61.5 (standard deviation 17.4; range 24.6-89.6; scale 0-100). Awareness of patients' perceptions of their QoL areas can enhance our understanding of an individual patient's QoL and reveal unsatisfactory areas where QoL could be improved with individually tailored needs based interventions. PMID- 19192078 TI - Self-esteem, negative emotionality, and depression as a common temperamental core: a study of mid-adolescent twin girls. AB - We tested the structure and magnitude of genetic and environmental influences on the overlap among self-esteem, negative emotionality, and major depression symptoms in adolescent girls (N=706) from the Minnesota Twin Family Study. Genetic and environmental influences on all three operated via a general, heritable factor. Genetic influences explained the majority of overlap among the three constructs, as well as most of the variance in self-esteem and negative emotionality. Genetic influences on depression were more modest and largely due to genetic factors specific to depression. These findings support the theory that self-esteem, depression, and neuroticism represent aspects of a common temperamental core. The interrelations among the three constructs in mid adolescence is consistent with their interrelations in adulthood. PMID- 19192081 TI - Views on nurse prescribing: a survey of community mental health nurses in the Republic of Ireland. AB - A nurse prescribing scheme has recently been implemented within the Republic of Ireland. This paper reports on the views of community mental health nurses on nurse prescribing just prior to the implementation of the scheme. Data were gathered through a 13-item questionnaire administered to 103 members of the Association of Community Mental Health Nurses in Ireland. Results indicated a distinct difference of view between male and female community mental health nurses, with female nurses having greater reservations towards the desirability of nurse prescribing in relation to educational preparation and impact on professional relationships. Overall, only 17% of respondents favoured being supervised in their prescribing practice by their consultant psychiatrist. The paper concludes that there is ambivalence towards prescribing in this important group of nurses which may need to be taken into account if nurse prescribing is to be successfully implemented within the Irish mental health service context. PMID- 19192082 TI - Exploring the past: mental health nursing in Greece. AB - This paper aims to explore the evolution of psychiatric nursing from the delayed organization of an asylum mental health system in Greece to the present. The care and custody of mental health patients which was in the hands of police at the first asylum passed to the hands of uneducated staff and gradually to the hands of qualified nurses. Severe overcrowding along with limited resources and low staff numbers resulted in the poor quality of living conditions. In this context, psychiatric nursing took its first steps in Greece, without realizing the inherent conflict in its dual role, that of care and social control. Counter to the handed-down cliche about psychiatric hospitals, nurses frequently put great effort into improving the living conditions of patients. However, the regulations of the institutions emphasizing the custodial role of nursing together with other structural factors of the psychiatric system impeded nurses' attempts to improve institutional care considerably. Nowadays, psychiatric hospitals are shrinking at a rapid pace, leaving behind them--as heritage for the psychiatric nurses who try to define their new role in community--the pioneers of psychiatric nursing's courage, ethos and respect for the psychiatric client. PMID- 19192083 TI - Nursing the disenfranchised: women who have relinquished an infant for adoption. AB - Grief that occurs as a result of relinquishing an infant for adoption is explored. Traditional grief models are cited as ineffectual for the satisfactory resolution of grief resulting from the relinquishment of a child for adoption. The reasons for disenfranchised grief are described and narratives of personal interviews provide insight into the grief process of birthmothers and evidence of their disenfranchisement. The role of psychiatric-mental health nurse is discussed and interventions aimed at assisting the birthmother to grieve are suggested. PMID- 19192084 TI - Patient health outcomes in psychiatric mental health nursing. AB - This integrative literature review examined evidence concerning the relationship between psychiatric mental health nursing interventions and patient-focused outcomes. Empirical studies, published between 1997 and 2007, were identified and gathered by searching relevant databases and specific data sources. Although 156 articles were critically appraised, only 25 of them met the inclusion criteria. Findings from this review showed that the most frequently used outcome instruments assessed psychiatric symptom severity. Most of the instruments targeted two symptom categories: altered thoughts/perceptions and altered mood. Other outcome instruments were categorized in the following domains: self-care, functioning, quality of life and satisfaction. The most important finding of this review is the lack of consistently strong evidence to support decisions concerning which outcome instrument or combination of instruments to recommend for routine use in practice. Based on this review, additional research to conceptualize, measure and examine the feasibility of outcome instruments sensitive to psychiatric mental health nursing interventions is recommended. PMID- 19192085 TI - Female caregivers' experiences of exposure to violence in nursing homes. AB - Although earlier studies have examined caregivers' experiences of exposure to violence, few have investigated female caregivers working in nursing homes with a specific focus on experiences throughout the entire scenario of a violent situation. This study illuminates female caregivers' experiences of being exposed to violence in nursing homes. Twenty caregivers working in three nursing homes located in northern Sweden were asked to narrate about a situation in which they had been exposed to violence. Their narratives were analysed by using qualitative content analysis. We found that the caregivers had preconceived ideas about violent behaviour, that they experienced a loss of control over the situation, and that they then strove to regain control. Experiences such as these may influence caregivers' interactions with residents who display violent behaviour. As a result of violent interactions, caregivers may distance themselves from the residents, an attitude that may decrease the quality of care. There is a risk that violence in nursing homes is accepted and normalized as a part of the job and hence persists. PMID- 19192086 TI - Varying views of life among people with long-term mental illness. AB - The purpose of this study was to explore views of life among people with long term mental illnesses. The participants' possible cognitive inability to express such views dictated a research design that was both fit for purpose and respectful of their integrity. The study, based on an ethnographic framework, involved photographs and interviews with five women and three men. The participants were the photographers, as well as the authors of their own narratives, and the photographs served as a starting point for the interviews. The interview material was analysed in terms of the phases of interpretation. Four main themes were identified: 'thoughts about God and the meaning of life and death', 'the meaning of relationships with others', 'how animals give meaning to life without demands' and 'the symbolic bearing of objects on life'. These four themes represent key existential issues among people with long-term mental illnesses, but they lack confidants to share or discuss these matters with. PMID- 19192087 TI - Mental health literacy of Australian Bachelor of Nursing students: a longitudinal study. AB - Many students have poor mental health literacy when they finish Bachelor of Nursing courses. This paper presents the findings of a longitudinal study of Australian Bachelor of Nursing students' mental health literacy about the effectiveness of interventions for people with schizophrenia. The 'Attitudes and Beliefs about Mental Health Problems: Professional and Public Views' questionnaire was used with a non-probability sample of nursing students. A time series approach to data collection was used, with data collected on three occasions between 2005 and 2007. Ethics approval was obtained from a university ethics committee. Data were analysed using SPSS Version 15.0. The students' views about the helpfulness of interventions showed a significant and positive improvement as they progressed through the course. There were significant differences over time in their views about the helpfulness of professional and lay interventions, their opinions about the helpfulness of mental health and other medications, and the usefulness of activity and non-pharmacological interventions. Because nursing students need to be mental health literate when they complete their course, mental health nursing content should be incorporated earlier in comprehensive undergraduate curricula and incrementally increased in each year of study. PMID- 19192088 TI - A narrative inquiry: moving on from homelessness for individuals with a major mental illness. AB - This narrative study explores the experience of 'moving on' from homelessness for individuals with major mental illnesses, after they have obtained permanent housing with supports. Twelve participants were interviewed up to three times over 6 months. There were various routes to homelessness, participants were homeless for varying lengths of time, and they described different journeys of 'moving on' in their lives. Place, and a series of places, were central for participants in this experience. The experience of homelessness for many could be described as 'on the move', in a circular pattern from shelter to shelter or street. Permanent housing and supports allowed participants to 'move on', reconnecting with family, getting jobs and planning for the future. Several participants wanted their stories used to send messages of hope, courage and survival. This study highlights the need for nurses to be aware of the concept of 'place' in the process of recovery from mental illness. PMID- 19192089 TI - The meaning of mental health nurses experience of providing one-to-one observations: a phenomenological study. AB - There has been much recent literature about the need for appropriate policies and approaches to ensure that patient's rights and standards of care are safeguarded. The focus from national policy is on suicide reduction and prevention, and the nursing literature has concentrated on the importance of engaging the patient and ensuring that there is the least amount of restriction possible. A research study was carried out to explore the perspective of mental health nurses working in these intensive situations, using a purposive sample of nursing staff from the local National Health Service Trust's acute units. A phenomenological approach to the study was chosen to allow an in-depth exploration of the issues--'seeing things up close'--using the philosophy of Husserl as a base. The key areas of enquiry were: to explore and amplify the experiences of nurses undertaking constant observations, including any effects that may have on nurses; to gain further understanding of the dynamics and processes involved; to discover information that can inform and support development needs. Individual taped interviews were carried out with six nurses working in an acute admission ward, and these were then transcribed and analysed using Giorgi's method of analysis. The results of this analysis showed that nurses are keenly aware of some of the professional and ethical tensions involved in the process of keeping patients safe while trying to promote recovery. Results have implications for policies, preparation and support of staff, and provide some further insights into the nature of this experience for nursing staff who are caring for patients who are presenting with risks to themselves. PMID- 19192090 TI - The convenient myth of Thomas Szasz. AB - Thomas Szasz's original critique of the concept of 'mental illness' is almost 50 years old. Over that half century Szasz has maintained a consistent campaign against the 'Therapeutic State', challenging the paternalism of coercive psychiatry and defending liberty and autonomy. Despite his widespread celebrity Szasz continues to be misread and misrepresented. In this paper we review some of Szasz's key ideas, in the light of Clarke's recent critique, setting this within the context of 'mental health nursing' and the problems in living affecting persons worldwide. PMID- 19192092 TI - Physical intervention: a review of the literature on its use, staff and patient views, and the impact of training. AB - As a principal control measure, physical intervention is intended to be a skilled manual, or hands-on, method of physical restraint implemented by trained individuals, with the intention of controlling the aggressive patient, to restore safety in the clinical environment. Physical intervention is however a contentious practice. There have been reports in the literature of negative psychological views from staff and patients on the procedure. Although formal structured training was introduced in response to concerns around patient safety during restraint, concerns remain that PI is sometimes construed as a stand-alone violence prevention initiative. Its potential for misuse, and overuse, in corrupted cultures of care has emerged as a social policy issue. The following paper critically explores the literature on training in physical intervention in the United Kingdom. PMID- 19192099 TI - Looking at movies and cartoons: eye-tracking evidence from Williams syndrome and autism. AB - BACKGROUND: Autism and Williams syndrome (WS) are neuro-developmental disorders associated with distinct social phenotypes. While individuals with autism show a lack of interest in socially important cues, individuals with WS often show increased interest in socially relevant information. METHODS: The current eye tracking study explores how individuals with WS and autism preferentially attend to social scenes and movie extracts containing human actors and cartoon characters. The proportion of gaze time spent fixating on faces, bodies and the scene background was investigated. RESULTS: While individuals with autism preferentially attended to characters' faces for less time than was typical, individuals with WS attended to the same regions for longer than typical. For individuals with autism atypical gaze behaviours extended across human actor and cartoon images or movies but for WS atypicalities were restricted to human actors. CONCLUSIONS: The reported gaze behaviours provide experimental evidence of the divergent social interests associated with autism and WS. PMID- 19192101 TI - Characteristics of African-American male caregivers in a study of oral health in Detroit--a brief communication. AB - OBJECTIVE: The role of fathers among African-American men, particularly related to oral health, has received relatively little scholarly attention. This paper describes the characteristics of African-American men who self-identified as primary caregiver to an index child participating in the Detroit Dental Health Project. METHODS: Of 1,021 caregiver-child pairs recruited to this oral health study, 52 were male. Data were collected at a central site in Detroit on: 1) demographics; 2) social support; 3) oral health beliefs, behaviors, and knowledge; 4) caregivers' and child's oral health. RESULTS: Participants reported good availability of social support and high perceived self-efficacy to take care of their child's teeth, yet, they possessed limited knowledge on preventing oral health problems. Moreover, male caregivers had high levels of caries, missing teeth, and poor hygiene. CONCLUSIONS: Findings may inform the development of effective interventions aimed at male caregivers to improve knowledge and understanding of the caries process, particularly concerning their children. PMID- 19192100 TI - Prevalence of early childhood caries among very young urban Boston children compared with US children. AB - OBJECTIVES: The aims of this study were to compare prevalence of early childhood caries (ECC) in 1- to 3-year-old children seeing primary-care pediatricians at two urban medical centers in Boston to the prevalence of ECC in similarly aged US children surveyed as part of the Third National Health and Nutrition Examination Survey (NHANES III) and to assess risk factors for ECC among this cohort of children compared with risk factors among similarly aged US children. METHODS: Characteristics of 787 1- to 3-year-old children from two urban Boston medical centers were compared with those of 3,644 similarly aged US children surveyed as part of NHANES III. Demographic and social characteristics and ECC prevalence by putative risk factors were compared. A multiple logistic regression model was fit to assess putative risk factors and difference between groups simultaneously. RESULTS: Race, age, previous dental visit, parents' education, and household income were significantly associated with ECC prevalence. Parents' place of birth was a significant effect modifier with lower ECC among Boston children of immigrants than among US children of immigrants. CONCLUSIONS: Lower ECC prevalence among urban Boston children of immigrant parents compared with US children of immigrant parents may reflect changing immigrant composition in the United States since NHANES III or a different immigrant composition in the Boston area compared with the United States. This finding reinforces the need for further research of immigrants in order to understand cultural practices that may affect oral health. Finally, low ECC prevalence among very young children reinforces the importance of early intervention in reducing ECC. PMID- 19192102 TI - Implementing a tobacco assisted referral program in dental practices. AB - OBJECTIVES: The objectives of this study are to design and implement a system level tobacco-control intervention in a large prepaid dental group practice and assess effects on staff performance measures and patient satisfaction. METHODS: We matched 14 dental facilities on size, socioeconomic status, smoking rate, and periodontal status, and then randomly assigned them to intervention or usual-care control. We trained intervention staff in an "Assisted Referral" team approach for assessing tobacco use, providing tailored advice and brief counseling, and encouraging smokers to talk by telephone with a specially trained tobacco counselor. Patients could call from the office or ask that the counselor call them later. Telephone counselors helped patients explore motivations and barriers for quitting; review available cessation-support strategies, programs, and medications; and identify next steps. RESULTS: During the 14-month study period, 66,516 members had annual- or new-patient examinations. Both intervention and control sites had high rates of tobacco assessment (97 percent) and advice (93 percent). Intervention patients were more likely than controls (69 percent versus 3 percent, P < 0.01) to receive additional chair-side tobacco counseling and assistance, and 11 percent agreed to receive additional telephone counseling. Intervention patients were more satisfied than controls with the dental team's tobacco-control efforts (P < 0.03). Referral rates varied substantially for different staff. CONCLUSIONS: The Assisted Referral approach was successfully integrated into routine dental care, was well received by patients, and resulted in increased patient satisfaction. Because free telephone-based tobacco counseling is now available nationwide, the approach may be a practical strategy for most dental-care settings. PMID- 19192103 TI - Assessing levels of agreement between two commonly used oral health-related quality of life measures. AB - OBJECTIVE: This study aimed to assess the level of agreement between two commonly used oral health-related quality of life (OHRQoL) measures, the short form of the Oral Health Impact Profile (OHIP14) and the Oral Impacts on Daily Performances (OIDP). METHODS: A sample of 1,675 15- to 16-year-old students attending all schools in Bauru (Sao Paulo, Brazil) was selected. The impact of oral conditions on quality of life in the last 6 months was reported using both OHIP14 and OIDP. To allow for comparison with the 100 percent OIDP score, OHIP14 scores were converted to percentages. Then, agreement between the two OHRQoL measures was analyzed using the Bland and Altman method. RESULTS: The mean difference between OHIP14 and OIDP was 6.48 percent [confidence interval 95% (6.08; 6.89)], with higher scores reported for OHIP14 than for OIDP. Besides, 95 percent of the differences between the two OHRQoL measures were between -10.59 and 23.56 percent. Finally, differences between OHIP14 and OIDP increased significantly as the magnitude of their average increased (P < 0.001). CONCLUSION: There was a moderate level of agreement between OHIP14 and OIDP, which may be partly due to the fact that both OHRQoL measures assess different levels of oral impacts on quality of life in addition to having different scoring systems. PMID- 19192105 TI - Changing inequalities in the distribution of caries associated with improving child oral health in Australia. AB - OBJECTIVES: This study aimed to document the changing distribution of and inequalities in dental caries in Australian children across the 25-year period from 1977 to 2002. METHODS: Oral health data were obtained from Australia's national Child Dental Health Survey Measures of caries distribution included the Significant Caries Index and the proportions of children with high caries experience [decayed, missing and filled teeth (DMFT) > or =4], while inequality was assessed by using Gini coefficients calculated from Lorenz curves. Changes in caries distribution were compared with changes in child dmft/DMFT. RESULTS: While appreciable reductions occurred in child caries experience, in terms of both mean dmft/DMFT and for those children with the poorest oral health, inequalities in the distribution of caries experience increased across the 25-year period. Inequalities in the distribution of decayed and filled teeth differed for the deciduous and permanent dentition and, in the permanent dentition, became increasingly similar in the 1990s. CONCLUSIONS: Increasing inequalities in child dental caries in Australia must be interpreted in the context of declines in both mean caries experience and in the caries experience of those children with the poorest oral health. The Gini coefficient documents that the majority of the caries experience is increasingly being confined to a smaller percentage of the child population; however, this is a consequence of population-wide child oral health improvements. PMID- 19192104 TI - Adequacy of oral health information for patients with diabetes. AB - OBJECTIVE: We investigated the perception of dental hygienists regarding their adequacy of providing diabetics with diabetes-related oral health preventive education. METHODS: A one-page questionnaire printed on both sides was mailed to 2,237 licensed registered dental hygienists with a South Carolina (SC) mailing address. In addition to the dental hygienists' background and practice characteristics, their perception of adequacy for educating patients with diabetes on various diabetes-related oral health topics and reasons for inadequate coverage of materials were queried in the survey. RESULTS: After two follow-up mailings, 995 completed and usable surveys were returned. An average of 93.6 percent of respondents indicated that they adequately covered topics of oral hygiene and general oral health issues. However, about 60 percent of respondents reported not covering all essential materials related to oral health when educating diabetic patients. The three most common reasons were: a) insufficient time (60.1 percent); b) patient disinterest (41.2 percent); and c) insufficient information on oral care and diabetes (39.7 percent). Respondents reporting insufficient information were less likely to adequately address the effect of periodontal disease on diabetes (P < 0.001), effect of uncontrolled diabetes on periodontal disease (P < 0.001), and dry mouth management (P = 0.03). CONCLUSION: This study indicates that SC dental hygienists do not routinely provide patient education on diabetes-related oral health and healthy lifestyle topics. Lack of time, patient disinterest, and insufficient information were the three main reasons for respondents not covering these essentials. A practical method for improving dental hygienists' comprehensive service to patients with diabetes is to offer them more continuing education on diabetes and oral health to supplement their knowledge, skills, and confidence to educate this growing population. PMID- 19192106 TI - Erectile dysfunction in type 2 diabetic men: relationship to exercise fitness and cardiovascular risk factors in the Look AHEAD trial. AB - INTRODUCTION: Determinants of erectile dysfunction in diabetic men have not been adequately investigated as potential mediators of change. AIM: To determine the prevalence and correlates of erectile dysfunction (ED) in overweight men with type 2 diabetes in the multicenter, Look AHEAD trial (Action for Health in Diabetes). MAIN OUTCOME MEASURES: International Index of Erectile Function (IIEF), self-reported use of phosphodiesterase type 5 inhibitors, laboratory measures of adiposity, cardiometabolic parameters, and exercise fitness. METHODS: Male participants aged 45-75 in the Look AHEAD trial in a committed relationship were recruited for an ongoing study of sexual function and diabetes. Eligible participants completed the IIEF questionnaire and provided updated information on use of medical treatments for sexual dysfunction. Baseline sexual function results for participants in the male ancillary study are reported here; intervention data and results for female participants are presented elsewhere. RESULTS: A total of 373 eligible male participants completed all sexual function questionnaires, of whom 263 (68.7%) were sexually active at the time of the study. Almost half (49.8%) of the men reported mild or moderate degrees of ED, and 24.8% had complete ED. Among sexually active participants, 42.6% had sought medical help for their problem, and 39.7% reported use of ED medications. ED was significantly associated with age (odds ratio [OR] = 1.05; confidence interval [CI]: 1.01-1.10) baseline HbA(1c) (OR = 1.31; CI: 1.05-1.63), hypertension history (OR = 2.41; CI: 1.34-4.36), and metabolic syndrome (OR = 3.05, CI: 1.31 7.11). Of note, cardiorespiratory fitness was found to be protective of ED in a multivariable analysis (OR = 0.61; P < 0.001). CONCLUSIONS: ED is prevalent in this sample of obese, type 2 diabetic men in the Look AHEAD study. Cardiovascular risk factors were highly associated with ED in this population, and cardiorespiratory fitness was protective in this analysis. PMID- 19192107 TI - Discordant fibrin formation in hemophilia. AB - BACKGROUND: The conversion of fibrinogen to fibrin and its crosslinking to form a stable clot are key events in providing effective hemostasis. OBJECTIVES: To evaluate the relationship of fibrinopeptide (FP) release and factor (F) XIII activation in whole blood from hemophiliacs. PATIENTS/METHODS: We investigated FPA and FPB release, FXIII activation and fibrin mass in tissue factor-initiated coagulation in whole blood from individuals with hemophilia and healthy subjects. RESULTS: In hemophiliacs, the rates of fibrin formation were delayed as compared to healthy individuals. FPA/FPB release and FXIII activation were decreased in hemophiliacs vs. healthy individuals: 5.4 +/- 0.7 microM min(-1) to 1.7 +/- 0.4 microM min(-1) (P = 0.003), 2.3 +/- 0.6 microM min(-1) to 0.5 +/- 0.1 microM min( 1) (P = 0.025), and 12.1 +/- 0.7 nM min(-1) to 3.1 +/- 0.7 nM min(-1) (P < 0.0005), respectively. More FPA was released in hemophiliacs (6.6 +/- 1.2 microM) prior to clot time (CT) than in healthy individuals (2.6 +/- 0.4 microM, P = 0.013), whereas FPB and activated FXIII levels remained comparable. FXIII activation, which normally coincides with FPA release, was delayed in hemophiliacs. At CT in normal blood, the FPA concentration was 2.6-fold higher than that of FPB (P = 0.003), whereas in hemophiliacs this ratio was increased to 6.6-fold (P = 0.001). CONCLUSIONS: These data suggest that essential dynamic correlations exist between the presentations of fibrin I, fibrin II, and FXIIIa. The 'discordance' of fibrin formation in hemophiliacs results in clots that are more soluble than normal (43% lower mass; P = 0.02). The resulting poor physical clot strength probably plays a crucial role in the pathology of hemophilia. PMID- 19192109 TI - p38 mitogen-activated protein kinase targets the production of proinflammatory endothelial microparticles. AB - BACKGROUND: Endothelial microparticles (EMPs) are irregularly shaped membrane fragments shed into the circulation in patients with vascular diseases, and may themselves act to enhance the endothelial response to inflammation. On the basis of the importance of p38 mitogen-activated protein kinase (MAPK) in endothelial responses to inflammatory stimuli, we sought to define the role of p38 in EMP generation and function. METHODS: Microparticle generation from cultures of human aortic endothelial cells (hAECs) treated with tumor necrosis factor-alpha (TNF alpha) and p38 inhibition was quantified via multiple modalities. The response of target endothelial cells was assessed by treatment of cells with EMPs generated under various conditions. RESULTS: Inhibition of p38 in hAECs, using pharmacologic agents, resulted in a 50% reduction of TNF-alpha-induced EMPs. Importantly, suppression of microparticles was specific to p38 MAPK pathways. EMPs triggered by TNF-alpha activation induced an approximately four-fold increase in soluble intercellular adhesion molecule-1 (sICAM-1) release from targeted cells. However, inhibition of p38 MAPK in the targeted cell prior to EMP treatment did not alter the sICAM1 response. CONCLUSIONS: Our findings implicate p38 MAPK signaling as significant and selective in the formation and maturation of EMPs. EMPs elicited a proinflammatory response from targeted hAECs that was dependent on the conditions under which EMPs were generated. However, our results imply a unidirectional model in which p38 MAPK is critical at the source of microparticle formation, but not the target cell response to EMPs. These findings indicate a novel mechanism by which p38 inhibition may offer therapeutic benefit in vivo via direct inhibition of EMP formation. PMID- 19192108 TI - Autoantibodies to heat shock protein 60 promote thrombus formation in a murine model of arterial thrombosis. AB - BACKGROUND AND OBJECTIVES: Anti-heat shock protein (HSP)60 autoantibodies are associated with atherosclerosis and are known to affect endothelial cells in vitro. However, their role in thrombus formation remains unclear. We hypothesized that anti-HSP60 autoantibodies could potentiate thrombosis, and evaluated the effect of anti-murine HSP60 antibodies in a ferric chloride (FeCl3)-induced murine model of carotid artery injury. METHODS: Anti-HSP60, or control, IgG was administered to BALB/c mice 48 h prior to inducing carotid artery injury, and blood flow was monitored using an ultrasound probe. RESULTS: Thrombus formation was more rapid and stable in anti-HSP60 IGG-treated mice than in controls (blood flow=1.7%+/-0.6% vs. 34%+/-12.6%, P=0.0157). Occlusion was complete in all anti HSP60 IgG-treated mice (13/13), with no reperfusion being observed. In contrast, 64% (9/14) of control mice had complete occlusion, with reperfusion occurring in 6/9 mice. Thrombi were significantly larger in anti-HSP60 IgG-treated mice (P=0.0001), and contained four-fold more inflammatory cells (P=0.0281) than in controls. Non-injured contralateral arteries of anti-HSP60 IgG-treated mice were also affected, exhibiting abnormal endothelial cell morphology and significantly greater von Willebrand factor (VWF) and P-selectin expression than control mice (P=0.0024 and P=0.001, respectively). CONCLUSIONS: In summary, the presence of circulating anti-HSP60 autoantibodies resulted in increased P-selectin and VWF expression and altered cell morphology in endothelial cells lining uninjured carotid arteries, and promoted thrombosis and inflammatory cell recruitment in FeCl3-injured carotid arteries. These findings suggest that anti-HSP60 autoantibodies may constitute an important prothrombotic risk factor in cardiovascular disease in human vascular disease. PMID- 19192111 TI - Molecular mechanism of the interaction between activated factor XIII and its glutamine donor peptide substrate. AB - BACKGROUND: Activated factor XIII (FXIII), a dimer of truncated A-subunits (FXIII A2*), is a transglutaminase that crosslinks primary amines to peptide-bound glutamine residues. Because in the few natural substrates of FXIII-A2* no consensus sequence could be identified around the reactive glutamine, studying the interaction between individual substrates and FXIII-A2* is of primary importance. Most of the alpha2-plasmin inhibitor (alpha2PI) molecules become truncated by a plasma protease, and the truncated isoform (N1-alpha2PI) is an important substrate of FXIII-A2*. The crosslinking of N1-alpha2PI to fibrin plays a major role in protecting fibrin from fibrinolysis. METHODS: We studied the interaction of FXIII-A2* with its dodecapeptide glutamine donor substrate, N1 alpha2PI(1-12), the sequence of which corresponds to the N-terminal sequence of N1-alpha2PI. Kinetic parameters for N1-alpha2PI(1-12) and for its truncated or synthetic mutants were determined by a spectrophotometric assay. The interaction of N1-alpha2PI(1-12) with FXIII-A2* was investigated by proton nuclear magnetic resonance (NMR) and saturating transfer difference (STD) NMR. RESULTS AND CONCLUSIONS: Kinetic experiments with peptides in which the Asn1 residue was either truncated or replaced by alanine and proton NMR analysis of the FXIII-A2* N1-alpha2PI(1-12) complex demonstrated that Asn1 is essential for effective enzyme-substrate interaction. Experiments with C-terminally truncated peptides proved that amino acids 7-12 are essential for the interaction of N1-alpha2PI(1 12) with the enzyme, and suggested the existence of a secondary binding site on FXIII-A2*. Hydrophobic residues, particularly Leu10 and the C-terminal Lys12, seemed to be especially important in this respect, and direct interaction between hydrophobic C-terminal residues and FXIII-A2* was demonstrated by STD NMR. PMID- 19192112 TI - Genetic alteration of the D2 domain abolishes von Willebrand factor multimerization and trafficking into storage. AB - BACKGROUND: The large von Willebrand factor (VWF) propeptide (VWFpp) plays a critical role in the multimerization and regulated storage of the mature VWF protein. Although our laboratory and others have identified mutations in von Willebrand disease patients that disrupt VWF multimerization, little is known about the affect of mutations on the regulated storage of VWF. PATIENTS/METHODS: We identified a heterozygous 18 base pair, in-frame deletion in exon 12 of the VWF gene in a patient with an unusual, dimer-intense multimer pattern. This deletion results in loss of amino acids 436-442 of VWFpp, which include one cysteine. RESULTS: Through expression studies, we demonstrate reduced secretion, loss of VWF multimerization, and defective regulated storage of the variant VWF. The loss of VWF storage is secondary to loss of propeptide storage resulting from an apparently defective sorting signal on VWFpp. Suprisingly, coexpressed wild type VWF or VWFpp functioned in trans to partially restore multimerization of VWF from the variant allele. CONCLUSIONS: The deletion of six amino acids in VWFpp results in defects in VWF processing, regulated storage, and function. Although VWFpp may usually function in a homotypic fashion, acting on its own mature VWF subunit, VWFpp may retain the ability to function in trans on VWF expressed from the variant allele. PMID- 19192114 TI - Hemophilia severity score system: validation from an Italian Regional Hemophilia Reference Center. PMID- 19192113 TI - Rap1b is critical for glycoprotein VI-mediated but not ADP receptor-mediated alpha2beta1 activation. AB - BACKGROUND: The platelet alpha2beta1 integrin functions as both an adhesion and signaling receptor upon exposure to collagen. Recent studies have indicated that alpha2beta1 function can be activated via inside-out signaling, similar to the prototypical platelet integrin alphaIIbbeta3. However, signaling molecules that regulate alpha2beta1 activation in platelets are not well defined. A strong candidate molecule is the small GTPase Rap1b, the dominant platelet isoform of Rap1, which regulates alphaIIbbeta3 activation. OBJECTIVES: We hypothesized that Rap1b positively regulates alpha2beta1 during agonist-induced platelet activation. METHODS: To test whether Rap1b activates alpha2beta1 downstream of glycoprotein (GP)VI or other platelet receptors, we stimulated platelets purified from Rap1b-/- or wild-type mice with diverse agonists and measured alpha2beta1 activation using fluorescein isothiocyanate-labeled monomeric collagen. We also examined the role of Rap1b in outside-in signaling pathways by analyzing adhesion and spreading of Rap1b-/- or wild-type platelets on monomeric, immobilized collagen. Finally, we monitored the activation status of related Rap GTPases to detect changes in signaling pathways potentially associated with Rap1b-mediated events. RESULTS: Rap1b-/- platelets displayed comparable ADP-induced or thrombin induced alpha2beta1 activation as wild-type platelets, but reduced convulxin dependent alpha2beta1 activation. Rap1b-/- platelets exhibited increased spreading on immobilized collagen but similar adhesion to immobilized collagen compared to wild-type platelets. Rap1b-/- platelets also showed Rap1a and Rap2 activation upon agonist stimulation, possibly revealing functional compensation among Rap family members. CONCLUSIONS: Rap1b is required for maximal GPVI-induced but not ADP-induced activation of alpha2beta1 in murine platelets. PMID- 19192115 TI - A new indication for statins to prevent venous thromboembolism? Not yet. PMID- 19192116 TI - Cancer begets venous thromboembolism, but is venous thromboembolism a risk factor for cancer? PMID- 19192117 TI - Underweight is associated with a reduced risk of venous thromboembolism. Results from the EDITH case-control study. PMID- 19192118 TI - Arterial cardiovascular events, statins, low-dose aspirin and subsequent risk of venous thromboembolism: a population-based case-control study. AB - BACKGROUND: Atherosclerotic disease has been associated with the risk of venous thromboembolism, but the available data are conflicting. There are similar confusions regarding the association of the use of aspirin and statins with venous thromboembolism. OBJECTIVES: To determine whether arterial cardiovascular events, use of statins and low-dose aspirin were associated with the risk of venous thromboembolism. PATIENTS AND METHODS: In this population-based case control study, we identified 5824 patients with venous thromboembolism and 58 240 population controls with a complete hospital and prescription history. We used logistic regression to estimate the relative risk of venous thromboembolism, adjusted for potentially confounding factors. RESULTS: Patients with a history of arterial cardiovascular events had a clearly increased relative risk. An event within 3 months before the index date conferred large increases in risk [relative risk 4.22 (95% confidence interval (CI), 2.33-7.64) after myocardial infarction, 4.41 (95% CI, 2.92-6.65) after stroke]. Myocardial infarction more than 3 months before the index date was not significantly associated with risk, although there was a relative risk of 1.29 (95% CI, 1.05-1.57) for myocardial infarction more than 60 months previously. A history of stroke was associated with small increases in risk after 3 months. Current use of statins was associated with a reduced risk of venous thromboembolism [relative risk=0.74 (95% CI, 0.63-0.85)]. Aspirin use was not associated with risk. CONCLUSIONS: Patients with cardiovascular events are at a short-term increased risk of venous thromboembolism. Statins might prevent venous thromboembolism but aspirin does not. However, as the study is non-randomized residual confounding cannot be excluded. PMID- 19192119 TI - The association of depression with platelet activation: evidence for a treatment effect. AB - BACKGROUND: Depression is associated with an increased risk of cardiovascular disease (CVD). Although the mechanism is uncertain, prothrombotic and inflammatory factors may play a role. OBJECTIVES: As platelets play a key role in CVD, we determined first, whether depressed individuals had more activated platelets than non-depressed individuals and second, whether treatment of depression reduced platelet activation levels. PATIENTS/METHODS: We recruited 108 depressed outpatients and 45 control subjects all without a history of CVD. After psychological assessment, the depressed patients were offered treatment with medication and/or psychotherapy. Flow cytometric markers of platelet activation and level of depression were assessed at baseline and at 4 weeks and 6 months after treatment. RESULTS: Depression was associated with increased platelet activation with a higher number of circulating CD62p (0.76x10(9) L(-1) vs. 0.46, P=0.019) and CD63 (P=0.05) positive platelets compared with controls. Patients with depression also had more circulating platelet-leukocyte aggregates than controls (P<0.001). There was a positive correlation between the severity of depression and the level of platelet activation. Platelets from depressed patients were also hyperreactive to adenosine 5 -diphosphate (ADP) stimulation with increased CD62p and CD63 exposure (P=0.003 and 0.019, respectively). Six months of treatment resulted in a reduced number of circulating CD62p and CD63 positive platelets (29.84% and 53.38% decrease) and a 20.9% reduction in CD63 exposure after ADP activation. CONCLUSIONS: Depression is associated with increased in vivo platelet activation and resolution of depression using psychotherapy and/or medication reduces platelet activation. These findings provide insights into the link between depression and cardiovascular risk. PMID- 19192120 TI - Evaluation of repatriation parameters: does medical history matter? AB - BACKGROUND: Aeromedical repatriation of patients is an expanding service due to the growing number of travelers worldwide. Of these repatriated patients, a small number require specialized transportation due to severe medical complications. We evaluated the medical in-flight records of Dutch patients with severe disease or polytrauma who were repatriated by air from 1998 to 2002 via one of the largest Dutch alarm centers. We questioned how this Dutch population of repatriated patients is demographically distributed and whether this population is a priori at high risk for acute medical complications that need specialized medical attention. RESULTS: Seventy-seven of 115 repatriated patients were 50 years and older, of which most were male (73%). Fifty patients had no significant medical history, whereas the remaining 65 patients suffered from comorbidities such as cardiovascular disease or cancer. In patients aged 18 to 49 years, one third of all patients were repatriated due to traumatic fractures. In the older age category, the main reasons for repatriation were cardiopulmonary incidents. There was an equal distribution in the primary medical reason for repatriation as defined by need for trauma/neurological support and ventilation or circulatory support. Of note, 82% of the 65 patients who traveled with a chronic disease condition were repatriated due to worsening of this particular condition. CONCLUSIONS: The present study shows that an aeromedical repatriation service is frequently employed by travelers with a history of chronic disease who develop medical complications. The growing number of repatriated elderly patients and/or patients with preexisting comorbidities requires development of secure pretravel risk assessment and adaptation of the medical service level in foreign countries. PMID- 19192121 TI - Online communication as a potential travel medicine research tool: analysis of messages posted on the TravelMed listserv. AB - BACKGROUND: Access to the Internet and electronic mail has created opportunities for online discussion that can facilitate medical education and clinical problem solving. Research into the use of these information technologies is increasing and the analysis of these tools can support and guide the activities of professional organizations, including educational endeavors. OBJECTIVE: The initial objective was to analyze patterns of information exchange on the International Society of Travel Medicine's (ISTM) travel health electronic mailing list related to a specific area of society interest. Secondary objectives included the analysis of listserv use in relation to subscriber demographics and rates of participation to support travel health educational activities. METHODS: This study examined the use of the ISTM TravelMed listserv over an 8-month period from January 1, 2006, to July 31, 2006. Descriptive data analysis included TravelMed user demographics, the type of posting, the topic and frequency of postings, and the source of information provided. RESULTS: During the study period, 911 (47%) of the eligible ISTM members subscribed to the TravelMed listserv. About 369 of these subscribers posted 1,710 individual messages. About 1,506 (88%) postings were educational; 207 (12%) postings were administrative. A total of 389 (26%) of the educational postings were primary queries and 1,120 (74%) were responses, with a mean string length of 2.9 responses per query (range: 1-51). Twenty participants contributed 40% of the educational postings. The topics with the most frequent postings were vaccines and vaccine-preventable diseases (473/31%) and malaria (258/17%). Postings focused on special populations, including pregnant women or immigrants, comprised a total of 14 postings (<1%). CONCLUSIONS: During the study period, a limited number of ISTM members (19%) authored postings on the listserv. Regular discussion centered on a limited number of recurring topics. The analysis provides several opportunities for the support of educational initiatives, clinical problem solving, and program evaluation. PMID- 19192122 TI - Rabies postexposure management of travelers presenting to travel health clinics in Auckland and Hamilton, New Zealand. AB - BACKGROUND: Rabies is a fatal disease, and travelers going to endemic areas need to take precautions. Little is known about the rabies postexposure management of travelers from New Zealand. METHODS: A total of 459 post-travel records from October 1998 until February 2006 at two travel medicine clinics, in Auckland and Hamilton, were examined for those who reported postexposure management to animals while abroad. RESULTS: Fifty-four patients were included, 48 (88.9%) were New Zealand residents and 52.0% were male. The mean age of exposed travelers was 30.4 years (SD = 15.5). There was an adult to child ratio of 5:1. The highest exposure risk group was those aged 16 to 30 years. South and Southeast Asia were the most prominent geographical regions where exposure occurred, with 45 (83.3%) of subjects being potentially exposed to rabies. Dogs were the commonest animals involved, accounting for two thirds of incidents (36; 66.7%). The commonest sites of animal exposure on the body were the thigh and lower leg (26; 48.1%) and the hand (10; 18.5%). Forty-six (85.2%) of the animal exposures were graded as World Health Organization (WHO) category III. Forty-nine (90.7%) of the travelers had not had preexposure prophylaxis. Once in New Zealand, the correct WHO postexposure prophylaxis regime was applied on 44 of 52 (84.6%) occasions. However, overall, only 25% of the sample received postexposure treatment consistent with WHO guidelines, reflecting inappropriate management abroad. CONCLUSIONS: Post-travel consultations at two New Zealand travel clinics were analyzed for prophylactic rabies postexposure management. The majority were travelers aged 16 to 30 years, who sustained WHO category III exposures to the lower limb in Asia, predominantly from dogs. Few of these travelers had been immunized prior to travel, and only 25% of them received postexposure prophylaxis consistent with WHO guidelines. Thus, 75% of the study sample remains at theoretical risk of contracting rabies due to inappropriate management overseas. PMID- 19192123 TI - Risk of hepatitis B for travelers: is vaccination for all travelers really necessary? AB - OBJECTIVES: Behavioral studies in travelers suggest that 33% to 76% of all travelers to hepatitis B virus (HBV)-endemic countries are at risk for HBV infection. We study the incidence and risk factors for HBV infection in travelers. METHODS: Retrospective analysis of the characteristics and risk factors of all reported acute HBV patients in Amsterdam, the Netherlands, from January 1, 1992, until December 31, 2003. RESULTS: The estimated incidence in travelers from Amsterdam to HBV-endemic countries is 4.5/100,000 travelers. Two thirds of these patients were immigrants who lived in Amsterdam and who had visited their friends and relatives in their country of origin. In 12 years, only three Dutch short-term tourists contracted HBV while traveling, all by heterosexual contacts. CONCLUSIONS: Dutch tourists who travel to HBV-endemic countries run a very low risk of contracting HBV. Vaccination of short-term Dutch tourists is not necessary. Immigrants run a higher risk irrespective of travel or duration of travel. This group should be advised vaccination. PMID- 19192124 TI - Search and rescue trends associated with recreational travel in US national parks. AB - BACKGROUND: National parks are popular travel destinations worldwide. Unfortunately, negative health experiences can and do occur during travel in national parks. This study investigated search and rescue (SAR) trends associated with recreational travel in US National Park Service (NPS) units. METHODS: A retrospective study was conducted of the Annual US National Park Service Search and Rescue Report for the years 2003 to 2006. RESULTS: From 2003 to 2006, there were 12,337 SAR operations involving 15,537 visitors. The total operational costs were US$16,552,053. The operations ended with 522 fatalities, 4,860 ill or injured visitors, and 2,855 saves. Almost half (40%) of the operations occurred on Saturday and Sunday, and visitors aged 20 to 29 years were involved in 23% of the incidents. Males accounted for 66.3% of the visitors requiring SAR assistance. Day hiking, motorized boating, swimming, overnight hiking, and nonmotorized boating were the participant activities resulting in the most SAR operations. The vast majority of visitors requiring SAR assistance were located within a 24-hour period, and the most common rescue environments were mountain areas between 1,524 and 4,572 m, lakes, rivers, oceans, and coastal areas. An error in judgment, fatigue and physical conditions, and insufficient equipment, clothing, and experience were the most common contributing factors. CONCLUSIONS: SAR incidents can be expensive and end with severe health consequences. NPS management should develop education and preventive efforts focused on hikers, boaters, and swimmers who are males and aged 20 to 29 years, addressing issues of adequate judgment, preparation, and experience. PMID- 19192125 TI - Cost-benefit of WC/rBS oral cholera vaccine for vaccination against ETEC-caused travelers' diarrhea. AB - BACKGROUND: The most common infectious health problem encountered by travelers to countries in the developing region is travelers' diarrhea (TD), with enterotoxigenic Escherichia coli (ETEC) being the most common pathogen isolated. Although mild in most cases, the disease still leads to the loss of a significant part of a vacation or business trip. There is currently a lack of knowledge about the costs in relation to the benefits of vaccination against TD caused by ETEC, and the purposes of this study were to estimate and develop a cost-benefit analysis of vaccination using whole-cell/recombinant-B-subunit oral cholera vaccine. METHODS: The consequences of the vaccination were identified and quantified in monetary terms. The cost-benefits for leisure and business travelers were assessed separately. The value of the travel was separated into the cost of the trip and of lost leisure time/business opportunities. A person with TD was in base case estimated to lose on average 3.5 days of a 7-day leisure trip and 2.5 days of a 4-day business trip. Results are presented for a Canadian traveler to endemic areas in year 2007 in US$. RESULTS: The average cost of a TD event was estimated at $1,460 and $1,996 for leisure and business travelers, respectively. The net value of the vaccination, however, varied with the risk of the disease. Through extensive literature searches, an updated ETEC map illustrating the proportion of ETEC-caused TD was created. CONCLUSIONS: The analysis indicated that vaccination would be considered cost-effective at incidence rates of ETEC-caused TD above about 13 and 9% for leisure and business travelers, respectively. It is, however, important to keep in mind that it is the value of the travel for the individual traveler that will decide if the vaccination provides good value for money. PMID- 19192126 TI - Description of the food safety system in hotels and how it compares with HACCP standards. AB - BACKGROUND: Tourism is an important earner of foreign exchange in Jamaica; hence, the protection of the visitors' health is very important. A study of travelers to Jamaica in 1996 to 1997 found that travelers' diarrhea (TD) affected almost 25% of visitors. The Ministry of Health (Jamaica) initiated a program for the prevention and control of TD aimed at reducing attack rates from 25.0% to 12.0% over a 5-year period through environmental health and food safety standards of hotels. This article examines the food safety systems in Jamaican hotels located in a popular resort area to find out how comparable they are with the Hazard Analysis Critical Control Point (HACCP) strategy. METHODS: A cross-sectional study was done of hotels in St. Mary and St. Ann. Quantitative data were obtained from food and beverage/sanitation staff and qualitative data through in-depth interviews with hotel managers. Observation of the food safety operations was also done. RESULTS: The majority (75%) of larger hotels used a combination of HACCP and Ministry of Health food safety strategies (p = 0.02) and offered all inclusive services (r =-0.705, p = 0.001). Larger hotels were more likely to have a better quality team approach, HACCP plan, and monitoring of critical control points (CCPs) and more likely to receive higher scores (p < 0.05). More than two thirds of hotel staff were knowledgeable of HACCP. Significantly smaller hotels (87.5%) received less than 70% in overall score (r = 0.75, p = 0.01). Identification of CCPs and monitoring of CCPs explained 96.6% of the change in the overall HACCP scores (p = 0.001). Hotel managers felt that some hotels' systems were comparable with HACCP and that larger properties were ready for mandatory implementation. Conclusions. While some components of the HACCP system were observed in larger hotels, there were serious shortcomings in its comparison. Mandatory implementation of HACCP would require that sector-specific policies be developed for smaller hotels and implemented on a phased basis. PMID- 19192127 TI - Evaluation of mood profiles during malaria chemoprophylaxis: a randomized, double blind, four-arm study. AB - BACKGROUND: To objectively compare the mood profiles of users of malaria chemoprophylaxis regimens (atovaquone-proguanil, chloroquine-proguanil, doxycycline, or mefloquine) in a group of nonimmune tourists to sub-Saharan Africa. METHODS: In a randomized, double-blind, four-arm study with placebo run in phase conducted at travel clinics in Switzerland, Germany, and Israel, we compared moods and feelings in chemoprophylaxis users (n= 547) by administering the standardized "Profile of Mood States" (POMS) questionnaire. This is designed to provide data on six categories of feelings: tension, depression, anger, vigor, fatigue, and confusion. The questionnaire was administered at four time points: recruitment (T1), 13 to 11 days before departure (T2), 6 to 4 days before departure (T3), and 7 to 14 days after return from Africa (T4). RESULTS: There were no significant differences with respect to overall mood impact between the medication arms. All scores were in the normal range, and no means were more than 1 SD from the norm. The POMS data were reanalyzed with respect to sex, age, medication group, and control time points (T1-T4). There were significant interaction effects between sex and medication group--women in the mefloquine group showed more "fatigue" (p= .011) and "confusion" (p= .011) than men. Significant effects of age group (below median age 34 y vs median age and above) were noted on the "tension" and "fatigue" scales in that less "tension" (p= .045) and less "fatigue" (p= .000) were noted in those aged 34 years and older. Younger participants, aged <34 years, reported more "confusion" (p= .013) at T2 than at T1 and T4. CONCLUSIONS: Although the overall mood profiles were similar for the users of any of the standard malaria chemoprophylaxis regimens, we found that women using mefloquine showed more fatigue and confusion than men and that younger persons aged less than 34 years, regardless of chemoprophylaxis used, reported more tension and fatigue than their older counterparts. PMID- 19192128 TI - Rethinking typhoid fever vaccines: implications for travelers and people living in highly endemic areas. PMID- 19192129 TI - Travelers' diarrhea in children visiting tropical countries. AB - We studied a group of 174 Portuguese children (aged 2 mo-16 y) who mostly traveled to tropical Portuguese-speaking countries and found an attack rate of 21.8% for travelers' diarrhea, much lower than previously described. We also showed that African rate analysis by region may hide significant differences between countries. PMID- 19192130 TI - Toxic hepatitis after consumption of traditional kava preparation. AB - Liver toxicity from the use of kava dietary supplements has been reported, but little is known about the side effects of traditional kava preparations. We present a case study of a tourist who developed serious toxic liver disease after consumption of kava beverages in traditional Samoan kava ceremonies. PMID- 19192131 TI - Fever, a blue hand, and abducens nerve paralysis in a returning traveler. AB - Fever, caused by Salmonella typhi, is the cause of significant morbidity worldwide. Extraintestinal manifestations of typhoid fever can confuse clinicians in Western countries, delaying diagnosis. We present an extraordinary case of typhoid fever, manifesting as hand cyanosis as well as abducens nerve paresis, all of which promptly resolved with antibiotics. PMID- 19192132 TI - Knowledge, attitudes, and practices evaluation about travel medicine in international travelers and medical students in Chile. AB - Because information about travel medicine in Chile is lacking, a knowledge, attitudes, and practices evaluation in international travelers and medical students was done. The travelers and medical students did not know the travel medicine and sanitary conditions of their destinations, although they perceived travel-associated health risks, but <10% had any vaccination and 5% got sick during international trips. PMID- 19192133 TI - Forty meals for a drop of blood ... AB - Increasing migration flow to Western countries poses formidable challenges from the epidemiological, clinical, and cultural standpoints. A case of Dhat syndrome is presented in a young Pakistani male migrant living in Italy, which required integrated medical and cultural approach to be solved after a through diagnostic workout that did not yield any result. PMID- 19192134 TI - Efficacy of antimalarial chemoprophylaxis for travelers. AB - Nine of 26 French tourists developed malaria after a 2-week stay under field conditions in a highly endemic place in Burkina Faso. A study of their preventive antimalarial measures identified a strong association of malaria attack with absence or inadequacy of chemoprophylaxis but not with mechanic measures. PMID- 19192136 TI - Acute hepatitis A in a young returning traveler from Kenya despite immunization before departure. AB - Aluminum-adsorbed hepatitis A vaccines are known to be highly efficient. We present here the case of a patient who was immunized against hepatitis A before leaving for Kenya and who contracted an acute symptomatic hepatitis A during travel. PMID- 19192135 TI - The Nepalese shepherd. AB - Neurocysticercosis is one of the most common causes of seizures in the developing world. Due to the high volumes of immigration from South America and Asia, American physicians are increasingly encountering this condition. This case report attempts to present a brief overview of some of the difficulties associated with the treatment of patients with a high disease burden. PMID- 19192137 TI - Melioidosis masquerading as community-acquired pneumonia: a case report demonstrating efficacy of intrapleural fibrinolytic therapy. AB - Burkholderia pseudomallei, a soil saprophyte typically associated with the highly fatal disease melioidosis in Southeast Asia, is an increasing global concern as worldwide travel proliferates in our increasingly cosmopolitan society. Emergence of this disease in locations where it was not previously described underscores the importance of understanding the basic microbiology, pathogenesis, and epidemiology of infectious diseases on a global scale rather than the myopic view of the usual microbiological culprits of community-acquired pneumonia (CAP) in developed nations. We report a case of melioidosis originally diagnosed as CAP complicated by empyema successfully cured with combination of antibiotics and intrapleural fibrinolytic therapy, averting operative intervention. PMID- 19192139 TI - Knowledge and use of measures to reduce health risks by corporate expatriate employees in western Ghana. PMID- 19192140 TI - Genetic and environmental risk indicators in canine non-Hodgkin's lymphomas: breed associations and geographic distribution of 608 cases diagnosed throughout France over 1 year. AB - BACKGROUND: The etiology of non-Hodgkin's lymphomas (NHL) is multifactorial. Environmental and genetic factors are frequently incriminated both in humans and dogs. OBJECTIVES: Our purpose was to study the geographic distribution of canine NHL (CNHL) in France and to evaluate genetic and environmental influences. ANIMALS: Six hundred and eight cases of CNHL, diagnosed throughout France over 1 year, were collected from 7 Veterinary Histopathologic Laboratories. METHODS: Retrospective study. Breeds affected by lymphomas were compared with the national population and associations between breed and immunophenotype were studied. The distribution of CNHL and canine T-cell NHL per 100,000 dogs per department was compared with the distribution of waste incinerators, polluted sites, and radioactive waste. RESULTS: The breeds significantly overrepresented among lymphoma cases were Boxer, Setter, and Cocker Spaniel (P < .001). There was a significant association between Boxer and T-cell NHL (P < .001), and between German Shepherd and Rottweiler and B-cell NHL (P < .01). The geographic distribution of CNHL and canine T-cell NHL indicated significant heterogeneity. Significant association between distributions of CNHL and waste incinerators (rho= 0.25, P < .05), polluted sites (rho= 0.36, P < .001), and radioactive waste (rho= 0.51, P < .001) was found. CONCLUSIONS AND CLINICAL IMPORTANCE: Influence of genetics in the development of CNHL was supported by the existence of an association between breed and immunophenotype. Waste incinerators, polluted sites, and radioactive waste could just be considered as risk indicators of CNHL, but not as risk factors. Case-control studies around critical sites are necessary to confirm the implication of those environmental factors in the development of CNHL. PMID- 19192141 TI - Incidence of transfusion reactions and retention of procoagulant and anticoagulant factor activities in equine plasma. AB - BACKGROUND: The extent of preservation of clotting factors and incidence of transfusion reactions to noncommercial equine plasma is not documented. HYPOTHESIS: Equine frozen plasma would retain its coagulation factor activity within the reference range and the incidence of transfusion reactions would be low. ANIMALS: Ten plasma donor horses. Fifty clinically ill hospitalized horses receiving plasma were reviewed to determine the incidence of reactions. METHODS: In vitro study and retrospective case review. Plasma was prepared by gravity sedimentation from whole blood refrigerated for 48 hours. The activities of factors VII through XII, antithrombin (AT), and Protein C were measured. Factor activities were compared for plasma samples obtained before blood collection (S0), after 48 hours of gravity sedimentation at 5 degrees C and after plasma separation (S1), and after 90 days of storage at -20 degrees C (S90). The medical records of 50 consecutive clinically ill horses receiving frozen plasma were reviewed to determine the incidence of transfusion reactions. RESULTS: The combined effect of plasma harvest, gravity sedimentation, decantation, and freezing caused significant reductions in factors IX, (43%P= .0013), X, (33%P= .0001), XI, (48%P= .0008), AT, (10%P= .02), and Protein C (26%P= .0001). Activities for all factors analyzed, except factor X, remained within the reference ranges. Transfusion reactions were recorded for 5/50 horses. CONCLUSIONS AND CLINICAL RELEVANCE: Clotting factors, AT, and Protein C were well preserved. The incidence of reactions to frozen plasma was 10%. PMID- 19192142 TI - Effect of parenteral selenium administration to sheep on prevalence and recovery from footrot. AB - BACKGROUND: Insufficient intake of selenium (Se) is common in many regions, and can contribute to increased susceptibility to and prolonged recovery from infectious diseases. OBJECTIVE: To determine the effect of Se administration in decreasing the severity and prevalence of footrot in sheep. ANIMALS: Thirty-eight footrot-affected and 19 nonaffected sheep from a commercial flock of known high incidence of footrot. METHODS: Placebo-controlled, prospective, 15-month clinical trial. Footrot-affected sheep were randomly assigned into 2 groups (n = 19) and injected with either 5 mg Se (footrot [FR]-Se) or saline (FR-Sal) at 1-month intervals for the duration of the study. Unaffected sheep (controls) received no treatment. Sheep feet were examined, trimmed, and scored for footrot with a scale of 0 (no footrot) to 4 (extensive) at 0, 3, 6, 9, and 15 months. Sheep were also bled at time 0 and then at 3, 6, and 15 months to assess whole blood Se concentrations. RESULTS: At time 0, control sheep (255 +/- 11 ng/mL) had higher (P < .05) whole blood Se concentrations compared with FR-Se (205 +/- 12 ng/mL) and FR-Sal (211 +/- 14 ng/mL) sheep. By 6 months, FR-Se sheep (317 +/- 9 ng/mL) had whole blood Se concentrations greater (P < .05) than both control (281 +/- 14 ng/mL) and FR-Sal (277 +/- 16 ng/mL) sheep. FR-Se ewes showed a faster decline in highest lesion score at 3 (P= .012) and 6 (P= .0036) months, and a greater decrease in the number of feet with foot score >0 at 6 (P= .020) months compared with FR-Sal ewes. Sheep with blood Se concentrations <300 ng/mL were at 3.5 times greater risk (1.1-12.1 confidence interval, odds ratio) for FR, although this relationship was only significant (P= .04) at 6 months of the study. CONCLUSIONS AND CLINICAL IMPORTANCE: In sheep with footrot, improved Se status in conjunction with routine control practices result in more rapid improvement of foot lesions. PMID- 19192143 TI - The effect of hetastarch (670/0.75) on urine specific gravity and osmolality in the dog. AB - BACKGROUND: Urine specific gravity (USG) is used clinically to estimate urine osmolality (UOsm). Although USG has been shown to have a linear correlation with UOsm in dogs, the relationship is altered when there are significant numbers of high molecular weight (MW) molecules in the urine. HYPOTHESIS: USG would no longer predict UOsm in dogs given intravenous hetastarch (670/0.75)(HES). ANIMALS: Eight healthy employee-owned adult dogs. METHODS: Prospective, controlled experimental study. USG and UOsm were measured every 30 minutes from t=0 minutes to t=360 minutes. Dogs were administered 20 mL/kg of either NaCl 0.9% (control group, n=4) or HES (treatment group, n=8) IV over 1 hour starting at t=90 minutes. RESULTS: There was a decrease in UOsm in both groups starting at t=120 minutes and continuing for the study duration, and there was no significant difference in UOsm between treatment and control groups across all time points. There was an appropriate decrease in USG from t=120 minutes for the control group. In the treatment group, USG increased significantly at t=120 minutes (P= .0006), t=150 minutes (P= .0002), and t=180 minutes (P= .0044). The largest increase in USG occurred at t=150 minutes with a mean USG of 1.070 +/- 0.021 (range 1.038-1.104). CONCLUSIONS AND CLINICAL IMPORTANCE: Urine specific gravity should not be used to estimate urine solute concentration in dogs following the administration of 20 mL/kg of HES. In a clinical setting, the evaluation of USG following this dose of HES may lead to an overestimation of urine concentration. PMID- 19192144 TI - Anti-endothelial cell antibodies in dogs with immune-mediated hemolytic anemia and other diseases associated with high risk of thromboembolism. AB - BACKGROUND: Dogs with immune-mediated hemolytic anemia (IMHA) and certain inflammatory diseases are at high risk of developing thromboembolic disease. The presence of anti-endothelial cell autoantibodies (AECA) has been associated with an increased risk of thromboembolism in humans. HYPOTHESIS: AECA will be detected more often in dogs at risk of thromboembolism than in healthy control animals or dogs with diseases not associated with a higher risk of thromboembolism. ANIMALS: Ninety-one sick dogs and 22 healthy control dogs. METHODS: Retrospective case controlled study. Serum was screened for the presence of AECA. Dogs were identified for the study based on the risk of thromboembolism as determined by clinical impression and the underlying disease process. Flow cytometry and normal canine endothelial cells were used to screen serum samples from sick and healthy control dogs for the presence of AECA. In addition, serum from dogs with confirmed thromboemboli was also screened for the presence of AECA by immunohistochemistry. RESULTS: AECA were detected in 2/91 sick dogs, both with infectious diseases, but were not found in healthy dogs. Anti-endothelial antibodies were not detected in 21 dogs with IMHA and 20 dogs with systemic inflammatory response syndrome, sepsis, or both. CONCLUSIONS: We conclude that AECA are rarely detectable in dogs considered at high risk of thromboembolism. These findings suggest that AECA may not play an important role in the pathogenesis of thromboembolism in dogs with IMHA and other inflammatory diseases. PMID- 19192145 TI - Peripheral nerve, neuromuscular and muscle disease. Abstracts of the 21st Annual Symposium of the European Society of Veterinary Neurology. September 25-27, 2008. Rhodes, Greece. PMID- 19192146 TI - Exogenous insulin treatment after hypofractionated radiotherapy in cats with diabetes mellitus and acromegaly. AB - BACKGROUND: The optimal treatment for feline acromegaly has yet to be established. Surgical and medical therapies are minimally effective although radiotherapy might have greater efficacy. The purpose of this study was to review the response and outcome of cats with acromegaly and insulin-resistant diabetes mellitus (DM) to radiotherapy. HYPOTHESES: That radiotherapy improves glycemic control in cats with acromegaly and that improved glycemic control is due to remission of clinical acromegaly; demonstrated by a fall in serum insulin-like growth factor-1 (IGF-1) concentrations. ANIMALS: Fourteen cats with naturally occurring acromegaly. METHODS: Retrospective case review; records of all cats treated for acromegaly with radiotherapy were reviewed from 1997 to 2008. Cats were selected on the basis of compatible clinical signs, laboratory features, and diagnostic imaging findings. Fourteen cats received radiotherapy, delivered in 10 fractions, 3 times a week to a total dose of 3,700 cGy. RESULTS: Thirteen of 14 cats had improved diabetic control after radiotherapy. These improvements were sustained for up to 60 months. DM progressed in 2 cats and 1 did not respond. Seven cats responded before the final treatment. Ten cats were euthanized, 1 as a consequence of radiotherapy. In 8 cats in which IGF-1 was measured after treatment, changes in its concentration did not reflect the clinical improvement in glycemic control. CONCLUSIONS AND CLINICAL IMPORTANCE: Radiotherapy represents an effective treatment for cats with insulin-resistant DM resulting from acromegaly. IGF-1 concentration after treatment does not provide a suitable method by which remission from either acromegaly or insulin-resistant DM may be assessed. PMID- 19192147 TI - Efficacy of combination chemotherapy for treatment of gastrointestinal lymphoma in dogs. AB - BACKGROUND: Chemotherapy for multicentric canine lymphoma has favorable results. The gastrointestinal (GI) tract is the most common extranodal site of canine lymphoma, but there have been no prospective studies to determine outcome when dogs with GI lymphoma are treated with chemotherapy. HYPOTHESIS: Treatment with a multiagent chemotherapy protocol is associated with a poor outcome in dogs with GI lymphoma. ANIMALS: Eighteen dogs with histologically confirmed GI lymphoma. METHODS: Prospective clinical trial in which dogs with GI lymphoma were treated with a 20-week combination chemotherapy protocol consisting of induction and consolidation phases. RESULTS: Thirteen dogs had primary GI lymphoma and 5 had multicentric lymphoma with GI involvement. The majority of the lymphomas (63%) were of T-cell origin. Overall remission rate was 56%; 9 dogs achieved a complete remission for a median of 86 days (range, 22-420 days) and 1 dog achieved a partial remission for 26 days. Overall median survival time was 77 days (range, 6 700 days). Dogs that failed to achieve a remission (10 versus 117 days; P= .002) or had diarrhea at initial presentation (70 versus 700 days; P < .001) had shorter survival times. CONCLUSION AND CLINICAL IMPORTANCE: The response and survival of dogs with GI lymphoma treated with multiagent chemotherapy is poor but long-term survival is possible. PMID- 19192148 TI - Concurrent Infection with Anaplasma phagocytophilum and Mycoplasma haemolamae in a young alpaca. PMID- 19192149 TI - Cellular and molecular characterization of a feline insulinoma. AB - BACKGROUND: Insulinoma is an autonomous insulin-secreting islet cell neoplasm that is rarely diagnosed in cats. The clinical and pathological aspects of feline insulinoma have been described previously, but the molecular characteristics of these tumors have not been investigated. OBJECTIVES: The study objectives were to characterize peptide hormone production and determine expression of selected genes involved in glucose metabolism and insulin secretion in a feline insulinoma. METHODS: Immunohistochemistry and RT-PCR were used to examine hormone and gene expression, respectively, by insulinoma cells. RESULTS: Immunohistochemistry examination indicated that the tumor cells expressed insulin, chromogranin A, and somatostatin but not glucagon or pancreatic polypeptide. The tumor expressed several genes characteristic of pancreatic beta cells (beta cells) including insulin (INS), glucose transporter 2 (GLUT2), and glucokinase (GCK). The tumor also expressed hexokinase 1 (HK1), a glycolytic enzyme not normally expressed in beta cells. GCK expression was higher in the insulinoma than in normal pancreas from the same cat. The GCK : HK1 ratio was >20 fold higher in insulinoma tissue than in normal pancreas. CONCLUSIONS AND CLINICAL IMPORTANCE: The feline insulinoma produced several peptide hormones and expressed genes consistent with a beta-cell phenotype. The pattern of hexokinase gene expression in tumor cells differed from that of normal pancreas. These findings suggest insulinoma cells may have an increased sensitivity to glucose that could contribute to the abnormal insulin secretory response observed at low serum glucose concentrations. PMID- 19192150 TI - Antemortem diagnosis of localized central nervous system histiocytic sarcoma in 2 dogs. PMID- 19192151 TI - Protozoal hepatitis associated with immunosuppressive therapy in a dog. PMID- 19192153 TI - Role of lung surfactant in respiratory disease: current knowledge in large animal medicine. AB - Lung surfactant is produced by type II alveolar cells as a mixture of phospholipids, surfactant proteins, and neutral lipids. Surfactant lowers alveolar surface tension and is crucial for the prevention of alveolar collapse. In addition, surfactant contributes to smaller airway patency and improves mucociliary clearance. Surfactant-specific proteins are part of the innate immune defense mechanisms of the lung. Lung surfactant alterations have been described in a number of respiratory diseases. Surfactant deficiency (quantitative deficit of surfactant) in premature animals causes neonatal respiratory distress syndrome. Surfactant dysfunction (qualitative changes in surfactant) has been implicated in the pathophysiology of acute respiratory distress syndrome and asthma. Analysis of surfactant from amniotic fluid allows assessment of fetal lung maturity (FLM) in the human fetus and exogenous surfactant replacement therapy is part of the standard care in premature human infants. In contrast to human medicine, use and success of FLM testing or surfactant replacement therapy remain limited in veterinary medicine. Lung surfactant has been studied in large animal models of human disease. However, only a few reports exist on lung surfactant alterations in naturally occurring respiratory disease in large animals. This article gives a general review on the role of lung surfactant in respiratory disease followed by an overview of our current knowledge on surfactant in large animal veterinary medicine. PMID- 19192152 TI - Hypothalamic-pituitary-adrenal axis assessment in healthy term neonatal foals utilizing a paired low dose/high dose ACTH stimulation test. AB - BACKGROUND: Hypothalamic-pituitary-adrenal (HPA) axis function is dynamic in the neonatal foal. The paired low dose/high dose cosyntropin (ACTH) stimulation test allows comprehensive HPA axis assessment, but has not been evaluated in neonatal foals. HYPOTHESIS: Foal age will significantly affect cortisol responses to a paired 10 and 100 microg dose cosyntropin stimulation test in healthy neonatal foals. ANIMALS: Twenty healthy neonatal foals. METHODS: HPA axis function was assessed in 12 foals at birth and at 12-24, 36-48 hours, and 5-7 days of age. At each age, basal cortisol and ACTH concentrations were measured and cortisol responses to 10 and 100 microg cosyntropin were assessed with a paired ACTH stimulation test protocol. Eight additional 36-48-hour-old foals received saline instead of 10 microg cosyntropin in the same-paired ACTH stimulation test design. RESULTS: At birth, foals had significantly higher basal cortisol and ACTH concentrations and higher basal ACTH : cortisol ratios compared with foals in all other age groups. A significant cortisol response to both the 10 and 100 microg doses of cosyntropin was observed in all foals. The magnitude of the cortisol response to both doses of cosyntropin was significantly different across age groups, with the most marked responses in younger foals. There was no effect of the paired ACTH stimulation test design itself on cortisol responses. CONCLUSIONS AND CLINICAL IMPORTANCE: A paired 10 and 100 microg cosyntropin stimulation test can be used to evaluate HPA axis function in neonatal foals. Consideration of foal age is important in interpretation of HPA axis assessment. PMID- 19192154 TI - Magnetic resonance imaging of right ventricular morphology and function in boxer dogs with arrhythmogenic right ventricular cardiomyopathy. AB - BACKGROUND: Arrhythmogenic right ventricular cardiomyopathy (ARVC) is a myocardial disease characterized by fibrofatty replacement of the right ventricle and ventricular tachyarrhythmias, reported most commonly in the Boxer dog. Although ARVC is characterized as a myocardial disease, the impact of the disease on the function of the right ventricle has not been well studied. OBJECTIVE: To noninvasively evaluate the function and anatomy of the right ventricle in Boxer dogs with ARVC. ANIMALS: Five adult Boxer dogs with ARVC and 5 healthy size matched hound dogs. METHODS: Magnetic resonance imaging was performed on an ECG gated conventional 1.5-T scanner using dark blood imaging and cine acquisitions. Images were evaluated by delineation of endocardial right and left ventricular contours in the end-diastolic and end-systolic phases of each slice. Right and left end-systolic and end-diastolic volumes were generated using Simpson's rule and ejection fraction was calculated. Images were evaluated for right ventricular (RV) aneurysms and wall motion abnormalities. Spin echo images were reviewed for the presence of RV myocardial fatty replacement or scar. RESULTS: RV ejection fraction was significantly lower in Boxers with ARVC compared with the controls (ARVC 34%+/- 11 control 53%+/- 10, P < .01). There was an RV aneurysm in 1 dog with ARVC but not in any of the controls. RV myocardial gross fatty changes were not observed in dogs of either group. CONCLUSIONS AND CLINICAL IMPORTANCE: These findings could be interpreted to suggest that arrhythmias and myocardial dysfunction precede the development of morphological abnormalities in dogs with ARVC. PMID- 19192155 TI - Unilateral and bilateral congenital sensorineural deafness in client-owned pure breed white cats. AB - BACKGROUND: Congenital sensorineural deafness has been reported frequently in experimental mixed-breed white cats but there is a paucity of data on occurrence of deafness in client-owned pure-breed white cats. OBJECTIVE: To describe hearing status in client-owned pure-breed white cats. ANIMALS: Eighty-four pure-breed client-owned cats with white coat color of 10 registered breeds presented for routine hearing evaluation before breeding (1995-2008). METHODS: Hearing was assessed by click-evoked brainstem auditory evoked response. RESULTS: Overall deafness prevalence was 20.2%; 9 cats (10.7%) were bilaterally deaf and 8 cats (9.5%) were unilaterally deaf. There was no association between sex and deafness status (P= .85). Deafness status was associated with iris color (P= .04). CONCLUSIONS AND CLINICAL IMPORTANCE: Congenital sensorineural deafness frequently occurs in pure-breed cats with white coat color. Unilateral sensorineural deafness was as common as bilateral deafness. PMID- 19192156 TI - Prevalence of deafness in dogs heterozygous or homozygous for the merle allele. AB - BACKGROUND: Deafness in dogs is frequently associated with the pigment genes piebald and merle. Little is known about the prevalence of deafness in dogs carrying the merle allele. OBJECTIVE: To determine the prevalence of deafness in dogs heterozygous and homozygous for the merle allele of the mouse Silver pigment locus homolog (SILV) gene. ANIMALS: One hundred and fifty-three privately owned merle dogs of different breeds and both sexes. METHODS: Hearing was tested by brainstem auditory-evoked response and classified as bilaterally hearing, unilaterally deaf, or bilaterally deaf. DNA from buccal cells was genotyped as either heterozygous or homozygous for the merle allele. Deafness association tests among merle genotype, eye color, and sex were performed by the chi(2) test. RESULTS: Deafness prevalence in merles overall was 4.6% unilaterally deaf and 4.6% bilaterally deaf. There was a significant association between hearing status and heterozygous versus homozygous merle genotype. For single merles (Mm), 2.7% were unilaterally deaf and 0.9% were bilaterally deaf. For double merles (MM), 10% were unilaterally deaf and 15% were bilaterally deaf. There was no significant association with eye color or sex. CONCLUSIONS: Deafness prevalence in merle dogs was greater than that in some dog breeds homozygous for the piebald gene, such as the English Cocker Spaniel, but comparable to, or lower than, that in the Dalmatian and white Bull Terrier. Dogs homozygous for the merle allele were significantly more likely to be deaf than heterozygotes. PMID- 19192157 TI - Validation of a commercially available immunoassay for the measurement of bovine cardiac troponin I. AB - BACKGROUND: Commercially available cardiac troponin I (cTnI) assays developed for use in humans have not yet been validated for use in cattle. HYPOTHESES: The ADVIA Centaur TnI-Ultra immunoassay can be used for the detection of bovine cTnI. In healthy cattle, serum cTnI is undetectable or is present only in trace amounts. METHODS: Purified bovine cTnI and cTnI-free bovine serum were used for the evaluation of assay performance including intra- and inter-assay precision, sensitivity, interference, linearity, and recovery. Effects of storage at 23, 4, 20, and -80 degrees C for 2 days, and at -20 and -80 degrees C for 7 and 14 days and repeated freeze-thaw cycles on recovery of cTnI were analyzed. Serum cTnI concentrations in 30 healthy dairy cows were determined. RESULTS: Intra- and inter-assay precisions (mean +/- SD) were 4.48 +/- 2.26 and 13.36 +/- 6.59%, respectively. The assay demonstrated linearity at 0.5, 2, 15, and 30 ng/mL cTnI. Mean recovery was 100.81, 85.26, 87.72, and 114.42%, respectively. Skeletal muscle homogenate added to serum of known cTnI concentration did not alter the concentration of the analyte (P > .05). Concentration of cTnI significantly decreased when samples were stored at 4 and 23 degrees C for 2 days (P < .05). Repeated freeze-thaw cycles and storage at -20 degrees C for 7 days had no significant influence on cTnI concentration (P > .05). Serum cTnI concentration in healthy cattle was 5 years. Eighty-five per cent of all families understood the information concerning head injury they had been given during the visit to the emergency department. However, only 69% received the information they needed about head injury in children before discharge from the emergency department. There were significant differences between the two age groups as to whether or not the information was addressed to the child or that the information was age appropriate. The results from our study should be interpreted with caution because of the relatively low number of respondents and the fact that the study was conducted using a questionnaire. Another limitation is the fact that we asked for the information 3 months posthead injury. CONCLUSIONS: Most families do understand the information that was provided and they also generally received the information they needed. However, they had not received information about common symptoms after a head injury. Strategies to improve information to families who experience a childhood head injury therefore seem necessary. PMID- 19192243 TI - The experience of caring in relatives to patients with serious mental illness: gender differences, health and functioning. AB - The aims of the study were twofold. First, the study examined the extent to which there are gender differences in the experience of caring in a sample of relatives of patients with serious mental illness. Secondly, the study examined the association of self-reports of health and functioning with the experience of burden among relatives of patients with serious mental illness. A total of 50 relatives of 32 patients receiving psychiatric treatment for their bipolar or psychotic condition completed the Experience of Caregiving Inventory (ECI) and COOP-WONCA (a self-report measure of general state of health and functioning). The study revealed that female relatives had a higher score on eight of 10 ECI subscores, and in both ECI negative and ECI positive. Additionally, the study revealed significant correlations between a higher score on the COOP-WONCA feeling subscore and the following ECI subscores: stigma (r = 0.304, p < 0.05), problem with services (r = 0.348, p < 0.05), need to back up (r = 0.335, p < 0.05), loss (r = 0.293, p < 0.05) and the total ECI negative subscore (r = 0.343, p < 0.05). We also found a significant correlation between a higher score on the overall health subscore (COOP-WONCA) and a higher score on problems with services (r = 0.290, p < 0.05). Gender was the strongest predictor of the ECI negative subscore. Neither the COOP-WONCA subscore physical health, daily activities or social activities correlated significantly with any of the ECI subscores. Our results showed strong associations between poor mental well-being and caregiver burden. We also found a higher association with gender and the feelings subscore (COOP-WONCA) on ECI negative than we had expected. PMID- 19192244 TI - Analyses of the recycling receptor, FcRn, in live cells reveal novel pathways for lysosomal delivery. AB - Lysosomes play a central role in the degradation of proteins and other macromolecules. The mechanisms by which receptors are transferred to lysosomes for constitutive degradation are poorly understood. We have analyzed the processes that lead to the lysosomal delivery of the Fc receptor, FcRn. These studies provide support for a novel pathway for receptor delivery. Specifically, unlike other receptors that enter intraluminal vesicles in late endosomes, FcRn is transferred from the limiting membrane of such endosomes to lysosomes, and is rapidly internalized into the lysosomal lumen. By contrast, LAMP-1 persists on the limiting membrane. Receptor transfer is mediated by tubular extensions from late endosomes to lysosomes, or by interactions of the two participating organelles in kiss-and-linger-like processes, whereas full fusion is rarely observed. The persistence of FcRn on the late endosomal limiting membrane, together with selective transfer to lysosomes, allows this receptor to undergo recycling or degradation. Consequently, late endosomes have functional plasticity, consistent with the presence of the Rab5 GTPase in discrete domains on these compartments. PMID- 19192245 TI - Calsyntenins mediate TGN exit of APP in a kinesin-1-dependent manner. AB - Kinesin motors are required for the export of membranous cargo from the trans Golgi network (TGN), yet information about how kinesins are recruited to forming transport intermediates is sparse. Here we show that the Kinesin-1 docking protein calsyntenin-1 localizes to the TGN in vivo and directly and specifically recruits Kinesin-1 to Golgi/TGN membranes as well as to dynamic post-Golgi carriers. Overexpression of various calsyntenin chimeras and kinesin light chain 1 (KLC1) at high levels caused the formation of aberrant membrane stacks at the endoplasmic reticulum (ER) or the Golgi, disrupted overall Golgi structure and blocked exit of calsyntenin from the TGN. Intriguingly, this blockade of calsyntenin exit strongly and selectively impeded TGN exit of amyloid precursor protein (APP). Using live cell microscopy we found that calsyntenins exit the TGN in Kinesin-1-decorated tubular structures which may serve as carriers for calsyntenin-1-mediated post-TGN transport of APP. Abrogation of this pathway via virus-mediated knockdown of calsyntenin-1 expression in primary cultured neurons caused a marked elevation of APP C-terminal fragments. Together, these results indicate a role for calsyntenin-1 in Kinesin-1-dependent TGN exit and post-Golgi transport of APP-containing organelles and further suggest that distinct intracellular routes may exhibit different capacities for proteolytic processing of APP. PMID- 19192246 TI - Basal body assembly in ciliates: the power of numbers. AB - Centrioles perform the dual functions of organizing both centrosomes and cilia. The biogenesis of nascent centrioles is an essential cellular event that is tightly coupled to the cell cycle so that each cell contains only two or four centrioles at any given point in the cell cycle. The assembly of centrioles and their analogs, basal bodies, is well characterized at the ultrastructural level whereby structural modules are built into a functional organelle. Genetic studies in model organisms combined with proteomic, bioinformatic and identifying ciliary disease gene orthologs have revealed a wealth of molecules requiring further analysis to determine their roles in centriole duplication, assembly and function. Nonetheless, at this stage, our understanding of how molecular components interact to build new centrioles and basal bodies is limited. The ciliates, Tetrahymena and Paramecium, historically have been the subject of cytological and genetic study of basal bodies. Recent advances in the ciliate genetic and molecular toolkit have placed these model organisms in a favorable position to study the molecular mechanisms of centriole and basal body assembly. PMID- 19192248 TI - Macroautophagy in immunity and tolerance. AB - Autophagy and proteasomal degradation constitute the two main catabolic pathways in cells. While the proteasome degrades primarily short-lived soluble proteins, macroautophagy, the main constitutive autophagic pathway, delivers cell organelles and protein aggregates for lysosomal degradation. Both the proteasome and macroautophagy are attractive effector mechanisms for the immune system because they can be used to degrade foreign substances, including pathogenic proteins, within cells. Therefore, both innate and adaptive immune responses use these pathways for intracellular clearance of pathogens as well as for presentation of pathogen fragments to the adaptive immune system. Because, however, the same mechanisms are used for the steady-state turnover of cellular self-components, the immune system has to be desensitized not to recognize these. Therefore, proteasomal degradation and macroautophagy are also involved in tolerizing the immune system prior to pathogen encounter. We will discuss recent advances in our understanding how macroautophagy selects self-structures in the steady state, how presentation of these on major histocompatibility complex class II molecules leads to tolerance and how macroautophagy assists both innate and adaptive immunity. This new knowledge on the specialized functions of the metabolic process macroautophagy in higher eukaryotes should allow us to target it for therapy development against immunopathologies and to improve vaccinations. PMID- 19192247 TI - Control of granule mobility and exocytosis by Ca2+ -dependent formation of F actin in pancreatic duct epithelial cells. AB - Elevation of intracellular Ca(2+) concentration ([Ca(2+)](i)) triggers exocytosis of secretory granules in pancreatic duct epithelia. In this study, we find that the signal also controls granule movement. Motions of fluorescently labeled granules stopped abruptly after a [Ca(2+)](i) increase, kinetically coincident with formation of filamentous actin (F-actin) in the whole cytoplasm. At high resolution, the new F-actin meshwork was so dense that cellular structures of granule size appeared physically trapped in it. Depolymerization of F-actin with latrunculin B blocked both the F-actin formation and the arrest of granules. Interestingly, when monitored with total internal reflection fluorescence microscopy, the immobilized granules still moved slowly and concertedly toward the plasma membrane. This group translocation was abolished by blockers of myosin. Exocytosis measured by microamperometry suggested that formation of a dense F-actin meshwork inhibited exocytosis at small Ca(2+) rises <1 microm. Larger [Ca(2+)](i) rises increased exocytosis because of the co-ordinate translocation of granules and fusion to the membrane. We propose that the Ca(2+) dependent freezing of granules filters out weak inputs but allows exocytosis under stronger inputs by controlling granule movements. PMID- 19192249 TI - Galectin-4-regulated delivery of glycoproteins to the brush border membrane of enterocyte-like cells. AB - We have previously reported that silencing of galectin-4 expression in polarized HT-29 cells perturbed apical biosynthetic trafficking and resulted in a phenotype similar to the inhibitor of glycosylation, 1-benzyl-2-acetamido-2-deoxy-beta-d galactopyranoside (GalNAcalpha-O-bn). We now present evidence of a lipid raft based galectin-4-dependent mechanism of apical delivery of glycoproteins in these cells. First, galectin-4 recruits the apical glycoproteins in detergent-resistant membranes (DRMs) because these glycoproteins were depleted in DRMs isolated from galectin-4-knockdown (KD) HT-29 5M12 cells. DRM-associated glycoproteins were identified as ligands for galectin-4. Structural analysis showed that DRMs were markedly enriched in a series of complex N-glycans in comparison to detergent soluble membranes. Second, in galectin-4-KD cells, the apical glycoproteins still exit the Golgi but accumulated inside the cells, showing that their recruitment within lipid rafts and their apical trafficking required the delivery of galectin 4 at a post-Golgi level. This lectin that is synthesized on free cytoplasmic ribosomes is externalized from HT-29 cells mostly in the apical medium and follows an apical endocytic-recycling pathway that is required for the apical biosynthetic pathway. Together, our data show that the pattern of N-glycosylation of glycoproteins serves as a recognition signal for endocytosed galectin-4, which drives the raft-dependent apical pathway of glycoproteins in enterocyte-like HT 29 cells. PMID- 19192251 TI - Centrosome function: sometimes less is more. AB - Tight regulation of centrosome duplication is critical to ensure that centrosome number doubles once and only once per cell cycle. Superimposed onto this centrosome duplication cycle is a functional centrosome cycle in which they alternate between phases of quiescence and robust microtubule (MT) nucleation and MT-anchoring activities. In vertebrate cycling cells, interphase centrioles accumulate less pericentriolar material (PCM), reducing their MT nucleation capacity. In mitosis, centrosomes mature, accumulating more PCM to increase their nucleation and anchoring capacities to form robust MT asters. Interestingly, functional cycles of centrosomes can be altered to suit the cell's needs. Some interphase centrosomes function as a microtubule-organizing center by increasing their ability to anchor MTs to form centrosomal radial arrays. Other interphase centrosomes maintain their MT nucleation capacity but reduce/eliminate their MT anchoring capacity. Recent work demonstrates that Drosophila cells take this to the extreme, whereby centrioles lose all detectable PCM during interphase, offering an explanation as to how centrosome-deficient flies develop to adulthood. Drosophila stem cells further modify the functional cycle by differentially regulating their two centrioles - a situation that seems important for stem cell asymmetric divisions, as misregulation of centrosome duplication in stem/progenitor cells can promote tumor formation. Here, we review recent findings that describe variations in the functional cycle of centrosomes. PMID- 19192250 TI - Glycosylation directs targeting and activation of cystatin f from intracellular and extracellular sources. AB - Cystatin F is a cysteine protease inhibitor that is selectively expressed in immune cells and unlike other cystatin family members is targeted to a significant extent to intracellular compartments. Initially made as an inactive glycosylated disulfide-linked dimer, cystatin F is converted to an active monomer by proteolytic cleavage following transport to the endosomal/lysosomal system. This active form of cystatin F targets cathepsin C/DPPI and probably other cathepsins in immune cells. We show that efficient targeting of cystatin F to the endocytic pathway is dependent not on its unique dimeric conformation but rather on its oligosaccharide chains. We demonstrate the unusual addition of N-linked sugars to an Asn-X-Cys motif in cystatin F and provide evidence that the mannose 6-phosphate sorting machinery is used to divert cystatin F from the secretory pathway and to mediate its uptake from extracellular pools. These studies identify a function for the oligosaccharides on cystatin F and raise the possibility that cystatin F might regulate proteases in trans by secretion in an inactive form by one cell and subsequent internalization and activation by another cell. PMID- 19192252 TI - A novel strategy for the invasive toxin: hijacking exosome-mediated intercellular trafficking. AB - Toxins penetrate mammalian cells through various means. In this study, we report a unique strategy used by trichosanthin (TCS), a plant toxin with ribosome inactivating activity, to penetrate host cells. We found that in both JAR and K562 cells, endocytosed TCS is incorporated into intraluminal vesicles of the multivesicular body (MVB) and is then secreted in association with these vesicles upon fusion of the MVB with the plasma membrane. The secreted TCS-loaded vesicles secreted by K562 cells move throughout the intercellular space and target syngeneic and specific allogeneic cells. Subsequent internalization permits delivery of the toxin into the cytosol, resulting in ribosomal inactivation and cell death. Thus, our findings provide a novel mechanism by which foreign proteins pass between and penetrate into mammalian cells. PMID- 19192253 TI - Defining macropinocytosis. AB - Macropinocytosis represents a distinct pathway of endocytosis in mammalian cells. This actin-driven endocytic process is not directly co-ordinated by the presence of cargo but can be induced upon activation of growth factor signalling pathways. The capacity to dissect the contribution of macropinocytosis to cellular processes has been hampered by a lack of unique molecular markers and defining features. While aspects of macropinosome formation and maturation are common to those shared by the other endocytic pathways, a number of key differences have recently begun to emerge and will be discussed in this study. It is now well established that macropinocytosis significantly contributes to antigen presentation by the immune system and is exploited by a range of pathogens for cellular invasion and avoidance of immune surveillance. PMID- 19192254 TI - Positive direct and indirect antiglobulin tests associated with oxaliplatin can be due to drug antibody and/or drug-induced nonimmunologic protein adsorption. AB - BACKGROUND: Two patients were suspected of having immune hemolytic anemia (IHA) due to oxaliplatin. A related drug, cisplatin, is known to cause nonimmunologic protein adsorption (NIPA). Studies were performed to determine the presence of oxaliplatin-dependent antibodies in addition to oxaliplatin-induced NIPA. STUDY DESIGN AND METHODS: Sera and eluates from the two patients were tested against red blood cells (RBCs) treated with oxaliplatin, cisplatin, and carboplatin (another platinum drug). Sera were also tested against untreated RBCs in the presence of the same drugs. Testing with pooled normal sera and anti-human albumin was used to demonstrate the presence of NIPA. Oxaliplatin-treated RBCs sensitized with the patients' sera and pooled normal sera were tested by a monocyte monolayer assay (MMA) to determine potential clinical significance. RESULTS: Both patients had high-titer antibodies to oxaliplatin in their sera that reacted with oxaliplatin-treated RBCs and with untreated RBCs in the presence of oxaliplatin. RBCs treated with oxaliplatin, cisplatin, and carboplatin all demonstrated NIPA (pooled normal sera and anti-human albumin were reactive to low titers). NIPA was also detected in tests with untreated RBCs in the presence of oxaliplatin and cisplatin. Lower-titer reactivity of both patients' sera with cisplatin may have been due to NIPA and/or cross-reactivity of anti-oxaliplatin with cisplatin. MMAs were weakly positive due to NIPA and more strongly positive due to oxaliplatin antibodies. CONCLUSION: Two patients with IHA were demonstrated to have oxaliplatin-dependent antibodies. Oxaliplatin was also shown to cause NIPA. The drug-dependent antibody and/or the drug-induced NIPA could have contributed to the patients' hemolytic anemia. PMID- 19192255 TI - A novel hepatitis B virus surface antigen immunoassay as sensitive as hepatitis B virus nucleic acid testing in detecting early infection. AB - BACKGROUND: The aim was to considerably enhance the sensitivity of hepatitis B virus (HBV) surface antigen (HBsAg) detection and investigate whether the window period for HBV detection could be reduced. STUDY DESIGN AND METHODS: A high sensitivity chemiluminescent enzyme immunoassay (CLEIA) was developed for quantitative HBsAg detection by a combination of monoclonal antibodies, each one for a specific epitope of HBsAg, and by improving the conjugation technique. The sensitivity of the assay was compared with that of the existing chemiluminescent immunoassay (CLIA). Commercially available seroconversion panels and samples of HBV-infected chimpanzees were tested with the developed prototype to assess whether the window period for HBsAg detection could be reduced to that for DNA detection. RESULTS: Compared to the existing CLIA, the CLEIA prototype detected HBsAg with approximately 230-fold higher sensitivity and showed a reduced window period. HBsAg detection by the CLEIA prototype and HBV DNA detection by polymerase chain reaction (PCR) occurred simultaneously. The mean time for the CLEIA prototype to first detect HBsAg was approximately 17.4 days less than that for the existing systems. Further, CLEIA prototype enabled HBsAg detection even in anti-HBs-positive seroconversion samples. In the inoculated chimpanzees the HBsAg and HBV DNA became detectable simultaneously and concentrations increased in parallel, whereas HBsAg remained detectable longer than HBV DNA in the declining phase of viremia. CONCLUSION: The CLEIA prototype yielded results comparable with those of HBV DNA PCR. This novel high-sensitivity assay may be useful for early detection of HBV infection and monitoring patients with a history of infection. PMID- 19192257 TI - Comparison of coagulation factor XIII content and concentration in cryoprecipitate and fresh-frozen plasma. AB - BACKGROUND: For patients with plasma coagulation factor XIII (pFXIII) deficiency, recommended means of replacement include infusions of fresh-frozen plasma (FFP), cryoprecipitate, or (where available) factor (F)XIII concentrates. Quantitative differences in pFXIII concentration in FFP and cryoprecipitate are not well defined and were, therefore, the subject of this study. STUDY DESIGN AND METHODS: FFP and cryoprecipitate (10 bags each from blood group O donors) were analyzed to quantify pFXIII activity and antigen. Coagulation FVIII, fibrinogen, and von Willebrand factor (VWF) were also quantitated. RESULTS: Mean (+/-SD) pFXIII activity in cryoprecipitate and FFP bags was 60 +/- 30 and 288 +/- 77 U per bag, respectively, and pFXIII antigen and activity levels were concordant. Other comparisons (mean +/- SD) between cryoprecipitate and FFP, respectively, were as follows: coagulation FVIII activity, 133 +/- 37 and 265 +/- 83 U per bag; fibrinogen content (Clauss kinetic assay), 183 +/- 44 and 725 +/- 199 mg per bag; VWF antigen content, 181 +/- 53 and 218 +/- 70 U per bag; VWF ristocetin cofactor activity, 168 +/- 34 and 221 +/- 65 U per bag; VWF collagen-binding activity, 164 +/- 40 and 208 +/- 71 U per bag; and fluid (plasma) volumes per bag, 21.3 +/- 2.7 and 245 +/- 29 mL. CONCLUSION: In contrast to other cryoprecipitable coagulation proteins, pFXIII is only mildly enriched in cryoprecipitate when compared with FFP (approx. two- to threefold). Although both products can provide effective pFXIII replacement, FFP may be preferred when infusion volume is not a major consideration and pFXIII concentrates are not available. VWF is substantially enriched in cryoprecipitate (approx. ninefold compared with its concentration in FFP), with VWF activity content exceeding that of FVIII by approximately 26 percent on average. PMID- 19192256 TI - JAL (RH48) blood group antigen: serologic observations. AB - BACKGROUND: JAL (RH48) is a low-prevalence antigen in the Rh blood group system and anti-JAL has caused hemolytic disease of the newborn. JAL is associated with either a haplotype carrying depressed C and e antigens or one carrying depressed c and e antigens. Blood samples from JAL+ people were tested, published serologic findings were confirmed, serologic studies were extended to include expression of other Rh antigens, and the antibody specificities produced by three sensitized JAL+ probands are reported. STUDY DESIGN AND METHODS: Red blood cell (RBC) samples from 17 (12 probands) JAL+ persons were tested by hemagglutination using standard methods. RESULTS: RBCs from both the Caucasian JAL+ probands had the (C)(e) haplotype and weakened C, e, hr(B), and hr(S) antigens. JAL+ samples from black persons had the (c)(e) haplotype and expressed weakened c, e, f, V, VS, hr(B), and hr(S) antigens. Plasma from three sensitized c+e+ JAL+ probands contained alloanti-c, alloanti-e, or alloantibody of apparent anti-Rh17 specificity. This study shows that this alloanti-Rh17-like antibody recognizes the high-prevalence antigen antithetical to JAL that has been named CEST. CONCLUSIONS: The presence of the JAL antigen has a quantitative (weakening) effect on the expression of C, e, hr(B), and hr(S) antigens in Caucasian persons and of c, e, f, V, VS, hr(B), and hr(S) antigens in people of black African ancestry. A qualitative effect also was demonstrated by the presence of alloanti c or alloanti-e in the plasma of two transfused c+e+ patients and by an antibody (anti-CEST) that recognizes the high-prevalence antigen antithetical to JAL. PMID- 19192258 TI - An economic evaluation of valsartan for post-MI patients in the UK who are not suitable for treatment with ACE inhibitors. AB - OBJECTIVES: The overall objective of this study was to estimate the costs and outcomes associated with treatment with valsartan for post-myocardial infarction (post-MI) patients with left ventricular systolic dysfunction, heart failure, or both, who are not suitable for treatment with angiotensin-converting enzyme (ACE) inhibitors, compared to placebo. METHODS: A Markov model, using data drawn from the Valsartan in Acute Myocardial Infarction (VALIANT) trial and other trials, was developed to predict the future health pathways, resource use, and costs for patients who have recently experienced an MI. Patients received either valsartan (mean dose 247 mg) or placebo. Cost data were drawn from national databases and published literature, although health outcome utility weights were derived from existing studies. Patient outcomes were modeled for 10 years, and incremental cost-effective ratios were calculated for valsartan compared with placebo. RESULTS: Over a period of 10 years, a cohort of 1000 patients treated with valsartan experienced 147 fewer cardiovascular deaths, 37 fewer nonfatal MIs, and 95 fewer cases of heart failure than a cohort who received placebo. The incremental cost of valsartan, compared with placebo, was 2680 pound per patient, although the incremental effectiveness of valsartan was 0.5021 quality-adjusted life-years (QALYs) gained per patient. Therefore, the incremental cost per QALY for treatment with valsartan was 5338 pound. When analysis was undertaken using life-years rather than QALYs, the cost per life-year gained was 4672 pound. CONCLUSIONS: For patients who are not suitable for treatment with ACE inhibitors, valsartan is a viable and cost-effective treatment for their management after an MI. PMID- 19192259 TI - Hypoactive sexual desire disorder in postmenopausal women: quality of life and health burden. AB - OBJECTIVES: To describe the health-related quality of life (HRQOL) implications of hypoactive sexual desire disorder (HSDD) in a national sample of postmenopausal women ages 30-70. METHODS: The Nationwide Survey of Female Sexual Health, a random-digit telephone survey of US households, collected information on female sexual function, demographic characteristics, HRQOL, and the presence of specific medical disorders from 1189 naturally or surgically postmenopausal women in stable relationships of >=3 months duration. HSDD was defined as <40 on the Profile of Female Sexual Function(c) scale and <60 on the Personal Distress Scale(c). Short Form-12 Health Survey (SF-12) summary and domain scores, and EuroQol (EQ-5D) index score and dimensions were compared with population-based norms for healthy individuals and selected chronic conditions. RESULTS: HSDD was associated with significant HRQOL decrements, with the largest SF-12 score differences in mental health (HSDD: 45.4 [standard error 1.9] vs. no HSDD: 51.0 [0.6], P < 0.01), vitality (HSDD: 47.7 [1.3] vs. no HSDD: 52.0 [0.7], P < 0.01), social function (HSDD: 47.3 [1.4] vs. no HSDD: 50.9 [0.7], P < 0.05), and bodily pain (HSDD: 41.4 [2.2] vs. no HSDD: 46.7 [0.9], P < 0.05). EQ-5D index was 0.08 points lower (HSDD: 0.76 [0.03] vs. no HSDD: 0.84 [0.02], P < 0.05) for those with HSDD compared with those without. HSDD was associated with a 0.1-point decrement in naturally menopausal women (HSDD: 0.78 [0.03] vs. no HSDD 0.88 [0.01], P < 0.01). Women with HSDD showed more HRQOL impairment than healthy population norms but were similar to adults with other chronic conditions such as diabetes and back pain. CONCLUSIONS: Women with HSDD showed substantial impairment in HRQOL. Given a prevalence of 6.6% to 12.5% among US women, HSDD represents an important burden on quality of life. PMID- 19192260 TI - Analytical validation of the Sysmex XT-2000iV for cell counts in canine and feline effusions and concordance with cytologic diagnosis. AB - BACKGROUND: The Sysmex XT-2000iV is a hematology analyzer that combines laser and impedance technology. Its usefulness for determining cell counts in canine and feline intracavitary effusions has not yet been studied. OBJECTIVES: The objectives of this study were to evaluate the analytical performance of the Sysmex XT-2000iV for cell counts in effusions from dogs and cats, and to assess correlation with an impedance counter and concordance with diagnoses based on cytologic findings. METHODS: Effusions (43 pleural, 23 peritoneal, 6 pericardial) were analyzed from 32 dogs and 34 cats. Total nucleated cell count (TNCC), HCT, and RBC count were determined on the Sysmex and compared with those obtained on an impedance counter (Hemat 8, SEAC). Imprecision, linearity, and limit of detection were determined for the Sysmex. An algorithm was designed using quantitative and qualitative data from the Sysmex to classify the effusions and the results were compared with diagnoses based on cytologic findings. RESULTS: Intra-assay and interassay coefficients of variation on the Sysmex were variable. Linearity of TNCC was >or=0.993 for dogs and cats, with the exception of effusions from cats with feline infectious peritonitis, which had delta (Delta) TNC values >3.0. In comparison with the Hemat 8, a proportional error was found for TNCC on the Sysmex. Effusion classification based on the algorithm was concordant with that obtained by cytologic examination in 43/72 (60%) samples. Discordant results usually were due to the misclassification of cells with similar morphology (such as mesothelial and carcinoma cells) in Sysmex scattergrams. CONCLUSION: The Sysmex XT-2000iV provides a precise and accurate TNCC and has moderate concordance with cytologic findings for classifying canine and feline effusions. Although microscopic examination of effusions is necessary to achieve an accurate diagnosis, the Sysmex can provide preliminary information that may be helpful to cytopathologists. PMID- 19192261 TI - Comparative study of hematologic and plasma biochemical variables in Eastern Atlantic juvenile and adult nesting loggerhead sea turtles (Caretta caretta). AB - BACKGROUND: Plasma biochemical and hematologic variables are important in the management of endangered sea turtles, such as loggerheads. However, studies on blood biochemistry and hematology of loggerheads are limited, and different concentrations according to variable criteria have been reported. OBJECTIVE: The purpose of this study was to establish and compare baseline plasma chemistry and hematology values in Eastern Atlantic juvenile and adult nesting loggerhead sea turtles (Caretta caretta). METHODS: Blood samples were collected from 69 healthy juvenile loggerhead sea turtles after their rehabilitation in captivity, and from 34 adult nesting loggerheads after oviposition. Fresh blood was used for leukocyte differential count and PCV determination. Heparinized blood was used for RBC and WBC counts. Plasma biochemical concentrations were measured using an automated biochemical analyzer. For the comparative study, nonparametric statistical analysis was done using the Mann-Whitney U-test. RESULTS: Minimum, maximum, and median concentrations were obtained for 14 hematologic and 15 plasma chemistry variables. Statistically significant differences between juvenile and adult turtles were found for PCV; RBC, WBC, and leukocyte differential counts; total protein, albumin, globulins, calcium, triglycerides, glucose, total cholesterol and urea concentrations; and lactate dehydrogenase activity. CONCLUSIONS: Age, size, and reproductive status cause important variations in the hematologic and plasma biochemical results of loggerheads. The reference values obtained in this study may be used as a standard profile, useful for veterinary surgeons involved in sea turtle conservation. PMID- 19192262 TI - Hematology and serum clinical chemistry reference intervals for free-ranging Scandinavian gray wolves (Canis lupus). AB - BACKGROUND: Scandinavian free-ranging wolves (Canis lupus) are endangered, such that laboratory data to assess their health status is increasingly important. Although wolves have been studied for decades, most biological information comes from captive animals. OBJECTIVES: The objective of the present study was to establish reference intervals for 30 clinical chemical and 8 hematologic analytes in Scandinavian free-ranging wolves. METHODS: All wolves were tracked and chemically immobilized from a helicopter before examination and blood sampling in the winter of 7 consecutive years (1998-2004). Seventy-nine blood samples were collected from 57 gray wolves, including 24 juveniles (24 samples), 17 adult females (25 samples), and 16 adult males (30 samples). Whole blood and serum samples were stored at refrigeration temperature for 1-3 days before hematologic analyses and for 1-5 days before serum biochemical analyses. Reference intervals were calculated as 95% confidence intervals except for juveniles where the minimum and maximum values were used. RESULTS: Significant differences were observed between adult and juvenile wolves for RBC parameters, alkaline phosphatase and amylase activities, and total protein, albumin, gamma-globulins, cholesterol, creatinine, calcium, chloride, magnesium, phosphate, and sodium concentrations. CONCLUSION: Compared with published reference values for captive wolves, reference intervals for free-ranging wolves reflected exercise activity associated with capture (higher creatine kinase activity, higher glucose concentration), and differences in nutritional status (higher urea concentration). PMID- 19192263 TI - Increased elastic microfibrils and thickening of fibroblastic nuclear lamina in canine cutaneous asthenia. AB - Cutaneous asthenia is a hereditary connective tissue disease, primarily of dogs and cats, resembling Ehlers-Danlos syndrome in man. Collagen dysplasia results in skin hyperextensibility, skin and vessel fragility, and poor wound healing. The purpose of this study was to describe the histological findings in a dog with a collagenopathy consistent with cutaneous asthenia. An 8-month-old crossbreed female dog presented with lacerations and numerous atrophic and irregular scars. The skin was hyperextensible and easily torn by the slightest trauma. Ultrastructurally, the dermis was comprised of elaunin and oxytalan microfibrils. These are immature fibres in which the fibrillar component is increased but elastin is reduced. Collagen fibres were profoundly disorganized. The fibrils had a highly irregular outline and a corroded appearance when viewed in cross section, and were spiralled and fragmented in a longitudinal view. Dermal fibroblasts displayed a conspicuous thickening of the nuclear lamina. Nuclear lamins form a fibrous nucleoskeletal network of intermediate-sized filaments underlying the inner nuclear membrane. Mutations in lamins or lamin-associated proteins cause a myriad of genetic diseases collectively called laminopathies. Disruption of the nuclear lamina seems to affect chromatin organization and transcriptional regulation of gene expression. A common link among all laminopathies may be a failure of stem cells to regenerate mesenchymal tissue. This could contribute to the connective tissue dysplasia seen in cutaneous asthenia. PMID- 19192264 TI - Predicting transfusions in cardiac surgery: the easier, the better: the Transfusion Risk and Clinical Knowledge score. AB - BACKGROUND AND OBJECTIVES: Allogeneic blood products transfusions are associated with an increased morbidity and mortality risk in cardiac surgery. At present, a few transfusion risk scores have been proposed for cardiac surgery patients. The present study is aimed to develop and validate a risk score based on adequate statistical analyses joint with a clinical selection of a limited (five) number of preoperative predictors. MATERIALS AND METHODS: The development series was composed of 8989 consecutive adult patients undergone cardiac surgery. Independent predictors of allogeneic blood transfusions were identified. Subsequently, five predictors were extracted as the most clinically relevant based on the judgement of 30 clinicians dealing with transfusions in cardiac surgery. A predictive score was developed and externally validated on a series of 2371 patients operated in another institution. The score was compared to the other existing scores. RESULTS: The following predictors constituted the Transfusion Risk and Clinical Knowledge score: age > 67 years; weight < 60 kg for females and < 85 kg for males preoperative haematocrit; gender--female; and complex surgery. At the external validation, this score demonstrated an acceptable predictive power (area under the curve 0.71) and a good calibration at the Hosmer-Lemeshow test. When compared to the other three existing risk scores, the Transfusion Risk and Clinical Knowledge score had comparable or better predictive power and calibration. CONCLUSION: A simple risk model based on five predictors only has a similar or better accuracy and calibration in predicting the transfusion rate in cardiac surgery than more complex models. PMID- 19192265 TI - Comparison of small n statistical tests of differential expression applied to microarrays. AB - BACKGROUND: DNA microarrays provide data for genome wide patterns of expression between observation classes. Microarray studies often have small samples sizes, however, due to cost constraints or specimen availability. This can lead to poor random error estimates and inaccurate statistical tests of differential expression. We compare the performance of the standard t-test, fold change, and four small n statistical test methods designed to circumvent these problems. We report results of various normalization methods for empirical microarray data and of various random error models for simulated data. RESULTS: Three Empirical Bayes methods (CyberT, BRB, and limma t-statistics) were the most effective statistical tests across simulated and both 2-colour cDNA and Affymetrix experimental data. The CyberT regularized t-statistic in particular was able to maintain expected false positive rates with simulated data showing high variances at low gene intensities, although at the cost of low true positive rates. The Local Pooled Error (LPE) test introduced a bias that lowered false positive rates below theoretically expected values and had lower power relative to the top performers. The standard two-sample t-test and fold change were also found to be sub-optimal for detecting differentially expressed genes. The generalized log transformation was shown to be beneficial in improving results with certain data sets, in particular high variance cDNA data. CONCLUSION: Pre-processing of data influences performance and the proper combination of pre-processing and statistical testing is necessary for obtaining the best results. All three Empirical Bayes methods assessed in our study are good choices for statistical tests for small n microarray studies for both Affymetrix and cDNA data. Choice of method for a particular study will depend on software and normalization preferences. PMID- 19192267 TI - Heterogeneous activation of the TGFbeta pathway in glioblastomas identified by gene expression-based classification using TGFbeta-responsive genes. AB - BACKGROUND: TGFbeta has emerged as an attractive target for the therapeutic intervention of glioblastomas. Aberrant TGFbeta overproduction in glioblastoma and other high-grade gliomas has been reported, however, to date, none of these reports has systematically examined the components of TGFbeta signaling to gain a comprehensive view of TGFbeta activation in large cohorts of human glioma patients. METHODS: TGFbeta activation in mammalian cells leads to a transcriptional program that typically affects 5-10% of the genes in the genome. To systematically examine the status of TGFbeta activation in high-grade glial tumors, we compiled a gene set of transcriptional response to TGFbeta stimulation from tissue culture and in vivo animal studies. These genes were used to examine the status of TGFbeta activation in high-grade gliomas including a large cohort of glioblastomas. Unsupervised and supervised classification analysis was performed in two independent, publicly available glioma microarray datasets. RESULTS: Unsupervised and supervised classification using the TGFbeta-responsive gene list in two independent glial tumor gene expression data sets revealed various levels of TGFbeta activation in these tumors. Among glioblastomas, one of the most devastating human cancers, two subgroups were identified that showed distinct TGFbeta activation patterns as measured from transcriptional responses. Approximately 62% of glioblastoma samples analyzed showed strong TGFbeta activation, while the rest showed a weak TGFbeta transcriptional response. CONCLUSION: Our findings suggest heterogeneous TGFbeta activation in glioblastomas, which may cause potential differences in responses to anti-TGFbeta therapies in these two distinct subgroups of glioblastomas patients. PMID- 19192266 TI - Fgf receptor 3 activation promotes selective growth and expansion of occipitotemporal cortex. AB - BACKGROUND: Fibroblast growth factors (Fgfs) are important regulators of cerebral cortex development. Fgf2, Fgf8 and Fgf17 promote growth and specification of rostromedial (frontoparietal) cortical areas. Recently, the function of Fgf15 in antagonizing Fgf8 in the rostral signaling center was also reported. However, regulation of caudal area formation by Fgf signaling remains unknown. RESULTS: In mutant mice with constitutive activation of Fgf receptor 3 (Fgfr3) in the forebrain, surface area of the caudolateral cortex was markedly expanded at early postnatal stage, while rostromedial surface area remained normal. Cortical thickness was also increased in caudal regions. The expression domain and levels of Fgf8, as well as overall patterning, were unchanged. In contrast, the changes in caudolateral surface area were associated with accelerated cell cycle in early stages of neurogenesis without an alteration of cell cycle exit. Moreover, a marked overproduction of intermediate neuronal progenitors was observed in later stages, indicating prolongation of neurogenesis. CONCLUSION: Activation of Fgfr3 selectively promotes growth of caudolateral (occipitotemporal) cortex. These observations support the 'radial unit' and 'radial amplification' hypotheses and may explain premature sulcation of the occipitotemporal cortex in thanatophoric dysplasia, a human FGFR3 disorder. Together with previous work, this study suggests that formation of rostral and caudal areas are differentially regulated by Fgf signaling in the cerebral cortex. PMID- 19192268 TI - Orthodontic tooth movement enhancing bony apposition in alveolar bony defect: a case report. AB - INTRODUCTION: Prevalence of complications from orthognathic surgery is relatively low but if it happens it is vital to manage the post complication bony defect appropriately. CASE PRESENTATION: This case report describes a 20-year-old gentleman who suffered from a complication from a bimaxillary orthognathic surgery. A bone grafting was carried out to repair the bony defect from the surgery but it was unsuccessful. A non-invasive technique employing the use of very light orthodontic force with a laceback stainless steel ligature is described and a successful space closure with an improvement in the periodontal condition and bone apposition has been shown. CONCLUSION: This technique can be considered if orthodontic tooth movement is needed across a deficient alveolar ridge. PMID- 19192269 TI - Human resources for health at the district level in Indonesia: the smoke and mirrors of decentralization. AB - BACKGROUND: In 2001 Indonesia embarked on a rapid decentralization of government finances and functions to district governments. One of the results is that government has less information about its most valuable resource, the people who provide the services. The objective of the work reported here is to determine the stock of human resources for health in 15 districts, their service status and primary place of work. It also assesses the effect of decentralization on management of human resources and the implications for the future. METHODS: We enumerated all health care providers (doctors, nurses and midwives), including information on their employment status and primary place of work, in each of 15 districts in Java. Data were collected by three teams, one for each province. RESULTS: Provider density (number of doctors, nurses and midwives/1000 population) was low by international standards--11 out of 15 districts had provider densities less than 1.0. Approximately half of all three professional groups were permanent public servants. Contractual employment was also important for both nurses and midwives. The private sector as the primary source of employment is most important for doctors (37% overall) and increasingly so for midwives (10%). For those employed in the public sector, two-thirds of doctors and nurses work in health centres, while most midwives are located at village level health facilities. CONCLUSION: In the health system established after Independence, the facilities established were staffed through a period of obligatory service for all new graduates in medicine, nursing and midwifery. The last elements of that staffing system ended in 2007 and the government has not been able to replace it. The private sector is expanding and, despite the fact that it will be of increasing importance in the coming decades, government information about providers in private practice is decreasing. Despite the promise of decentralization to increase sectoral "decision space" at the district level, the central government now has control over essentially all public sector health staff at the district level, marking a return to the situation of 20 years ago. At the same time, Indonesia has changed dramatically. The challenge now is to envision a new health system that takes account of these changes. Envisioning the new system is a crucial first step for development of a human resources policy which, in turn, will require more information about health care providers, public and private, and increased capacity for human resource planning. PMID- 19192270 TI - Dental root canal treatment complicated by foreign body ingestion: a case report. AB - INTRODUCTION: Most foreign bodies pass through the gastrointestinal tract uneventfully. The majority of the reported literature describes the management of ingested blunt objects. However, ingestion of sharp objects can still occur with a higher rate of perforation corresponding to treatment dilemmas. CASE PRESENTATION: We report the conservative management of an inadvertently ingested sharp foreign body during a routine dental procedure and describe a management strategy for the treatment of both blunt and sharp foreign bodies. CONCLUSION: Urgent endoscopic assessment and retrieval is indicated when there is a history of a recently ingested sharp foreign body or if clinical suspicion suggests that the object is located within the oesophagus. Conservative management is advocated if the object has passed through the pylorus with serial clinical assessments including daily radiographs. Surgical intervention is warranted in the presence of obstruction, perforation or peritonitis. PMID- 19192271 TI - Currarino's syndrome misinterpreted as Hirschsprung's disease for 17 years: a case report. AB - BACKGROUND: Currarino's syndrome is an autosomal dominant hereditary disease known by the triad of anorectal stenosis, anterior sacral defect, and a presacral mass that is most often an anterior sacral meningocele. Actually this syndrome could remain asymptomatic in many instances, but symptomatic patients might present with constipation as the sole manifestation of the Currarino's syndrome among the other wide spectrum of manifestations. CASE PRESENTATION: An 18-year old woman was diagnosed with a late-recognized Currarino syndrome, presented by a longstanding constipation which had been wrongly diagnosed and treated as Hirschsprung's disease since early childhood. CONCLUSION: Long-lasting constipation could imply to neural tube anomaly such as anterior sacral meningocele with or without association to Currarino's syndrome. PMID- 19192272 TI - Redistribution of joint moments is associated with changed plantar pressure in diabetic polyneuropathy. AB - BACKGROUND: Patients with diabetic polyneuropathy (DPN) are often confronted with ulceration of foot soles. Increased plantar pressure under the forefoot has been identified as a major risk factor for ulceration. This study sets out to test the hypothesis that changes in gait characteristics induced by DPN related muscle weakness are the origin of the elevated plantar pressures. METHODS: Three groups of subjects participated: people diagnosed with diabetes without polyneuropathy (DC), people diagnosed with diabetic polyneuropathy (DPN) and healthy, age matched controls (HC). In all subjects isometric strength of plantar and dorsal flexors was assessed. Moreover, joint moments at ankle, knee and hip joints were determined while walking barefoot at a velocity of 1.4 m/s. Simultaneously plantar pressure patterns were measured. RESULTS: Compared to HC-subjects, DPN participants walked with a significantly increased internal plantar flexor moment at the first half of the stance phase. Also in DPN-subjects the maximal braking and propelling force applied to the floor was decreased. Moreover, in DPN subjects the ratio of forefoot-to-rear foot plantar pressures was increased. Body mass normalized strength of dorsal flexors showed a trend to be reduced in people with diabetes, both DC and DPN, compared to HC-subjects. Plantar flexors tended to be less weak in DC compared to HC and in DPN relative to DC. CONCLUSION: The results of this study suggest that adverse plantar pressure patterns are associated with redistribution of joint moments, and a consequent reduced capacity to control forward velocity at heel strike. PMID- 19192273 TI - Identification of genes differentially expressed between benign and osteopontin transformed rat mammary epithelial cells. AB - BACKGROUND: Osteopontin is a secreted, integrin-binding and phosphorylated acidic glycoprotein which has an important role in tumor progression. FINDINGS: In this study, we have utilized suppressive subtractive hybridization (SSH) to evaluate OPN regulated gene expression, using the Rama 37 benign non-invasive rat mammary cell line and a subclone, Rama 37-OPN. Rama 37-OPN was produced by stably transfecting Rama 37 with an OPN expression vector and it demonstrates increased malignant properties in vitro. Sequence and expression array analysis of the respective cDNA libraries of over 1600 subtracted cDNA fragments revealed 982 ESTs, 45 novel sequences and 659 known genes. The known up-regulated genes in the Rama 37-OPN library code for proteins with a variety of functions including those involved in metabolism, cell adhesion and migration, signal transduction and in apoptosis. Four of the most differentially expressed genes between the benign and in vitro malignant rat mammary cell lines are tumor protein translationally controlled I (TPTI), aryl hydrocarbon receptor nuclear translocator (ARNT), ataxia telangiectasia mutated (ATM) and RAN GTPase (RAN). The largest difference (ca 10,000 fold) between the less aggressively (MCF-7, ZR-75) and more aggressively malignant (MDA MB 231, MDA MB 435S) human breast cancer cell lines is that due to RAN, the next is that due to osteopontin itself. CONCLUSION: The results suggest that enhanced properties associated with the malignant state in vitro induced by osteopontin may be due to, in part, overexpression of RAN GTPase and these biological results are the subject of a subsequent publication 1. PMID- 19192275 TI - Eradication of multidrug-resistant Acinetobacter baumannii in a female patient with total hip arthroplasty, with debridement and retention: a case report. AB - INTRODUCTION: Multidrug-resistant Acinetobacter baumannii has become a significant cause of healthcare-associated infections, but few reports have addressed Acinetobacter baumannii infections associated with orthopedic devices. The current recommended treatment for complicated infections due to orthopedic devices, including resistant gram-negative rods, consists of antimicrobial therapy with debridement and removal of implants. CASE PRESENTATION: The patient, a 47-year-old woman, had previously had a prior total hip arthroplasty at 16 years of age for a complex femoral neck fracture, and multiple subsequent revisions. This time, she underwent a fifth revision secondary to pain. Surgery was complicated by hypotension resulting in transfer to the intensive care unit and prolonged respiratory failure. She received peri-operative cefazolin but postoperatively developed surgical wound drainage requiring debridement of a hematoma. Cultures of this grew ampicillin-sensitive Enterococcus and Acinetobacter baumannii (sensitive only to amikacin and imipenem). The patient was started on imipenem. Removal of the total hip arthroplasty was not recommended because of the recent surgical complications, and the patient was eventually discharged home. She was seen weekly for laboratory tests and examinations and, after 4 months of therapy, the imipenem was discontinued. She did well clinically for 7 months before recurrent pain led to removal of the total hip arthroplasty. Intra-operative cultures grew ampicillin-sensitive Enterococcus and coagulase-negative Staphylococcus but no multidrug-resistant Acinetobacter baumannii. The patient received ampicillin for 8 weeks and had not had recurrent infection at the time of writing, 37 months after discontinuing imipenem. CONCLUSION: We describe the successful treatment of an acute infection from multidrug-resistant Acinetobacter baumannii with debridement and retention of the total hip arthroplasty, using monotherapy with imipenem. This case challenges the general assumption that all orthopedic-device infections due to multidrug-resistant gram-negative organisms will require hardware removal. Further studies are needed to determine if organisms such as multidrug-resistant Acinetobacter baumannii are amenable to treatment with hardware retention. PMID- 19192274 TI - Antirheumatic drug response signatures in human chondrocytes: potential molecular targets to stimulate cartilage regeneration. AB - INTRODUCTION: Rheumatoid arthritis (RA) leads to progressive destruction of articular cartilage. This study aimed to disclose major mechanisms of antirheumatic drug action on human chondrocytes and to reveal marker and pharmacological target genes that are involved in cartilage dysfunction and regeneration. METHODS: An interactive in vitro cultivation system composed of human chondrocyte alginate cultures and conditioned supernatant of SV40 T-antigen immortalised human synovial fibroblasts was used. Chondrocyte alginate cultures were stimulated with supernatant of RA synovial fibroblasts, of healthy donor synovial fibroblasts, and of RA synovial fibroblasts that have been antirheumatically treated with disease-modifying antirheumatic drugs (DMARDs) (azathioprine, gold sodium thiomalate, chloroquine phosphate, and methotrexate), nonsteroidal anti-inflammatory drugs (NSAIDs) (piroxicam and diclofenac), or steroidal anti-inflammatory drugs (SAIDs) (methylprednisolone and prednisolone). Chondrocyte gene expression profile was analysed using microarrays. Real-time reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay were performed for validation of microarray data. RESULTS: Genome-wide expression analysis revealed 110 RA-related genes in human chondrocytes: expression of catabolic mediators (inflammation, cytokines/chemokines, and matrix degradation) was induced, and expression of anabolic mediators (matrix synthesis and proliferation/differentiation) was repressed. Potential marker genes to define and influence cartilage/chondrocyte integrity and regeneration were determined and include already established genes (COX-2, CXCR-4, IL-1RN, IL-6/8, MMP-10/12, and TLR-2) and novel genes (ADORA2A, BCL2-A1, CTGF, CXCR-7, CYR-61, HSD11B-1, IL-23A, MARCKS, MXRA-5, NDUFA4L2, NR4A3, SMS, STS, TNFAIP-2, and TXNIP). Antirheumatic treatment with SAIDs showed complete and strong reversion of RA-related gene expression in human chondrocytes, whereas treatment with NSAIDs and the DMARD chloroquine phosphate had only moderate to minor effects. Treatment with the DMARDs azathioprine, gold sodium thiomalate, and methotrexate efficiently reverted chondrocyte RA-related gene expression toward the 'healthy' level. Pathways of cytokine-cytokine receptor interaction, transforming growth factor-beta/Toll-like receptor/Jak-STAT (signal transducer and activator of transcription) signalling and extracellular matrix receptor interaction were targeted by antirheumatics. CONCLUSIONS: Our findings indicate that RA-relevant stimuli result in the molecular activation of catabolic and inflammatory processes in human chondrocytes that are reverted by antirheumatic treatment. Candidate genes that evolved in this study for new therapeutic approaches include suppression of specific immune responses (COX-2, IL-23A, and IL-6) and activation of cartilage regeneration (CTGF and CYR-61). PMID- 19192277 TI - Amoebic liver abscess - a cause of acute respiratory distress in an infant: a case report. AB - INTRODUCTION: The usual presentation of amebic liver abscess in children is extremely variable and unpredictable. It presents with a picture of common pediatric illness that is fever, lethargy, and abdominal pain, and can go on to develop into a rare complication of rupture into the pleura to cause acute respiratory distress, which is another common pediatric illness. In our patient, diagnosis was not made or suspected in these two stages. CASE PRESENTATION: This is the report of a 2-year-old male infant who presented with a 2-week history of anorexia, fever, and abdominal pain. A few hours after admission, he suddenly developed acute respiratory distress; chest X-ray demonstrated massive right pleural effusion that failed to response to tube thoracostomy. Limited thoracotomy revealed a ruptured amebic liver abscess through the right cupola of the diaphragm. The content of the abscess was evacuated from the pleural cavity, which was drained with two large chest tubes. Serological examination confirmed the diagnosis of ruptured amebic liver abscess. Postoperative treatment with antibiotics including metronidazole continued until full recovery. CONCLUSION: Diagnosis of such a rare disease requires a high degree of suspicion. In this patient, the diagnosis was only made postoperatively. The delay in presentation and the sudden onset of respiratory distress must be emphasized for all those physicians who care for children. PMID- 19192276 TI - Calcitonin gene-related peptide stimulates proliferation of alveolar epithelial cells. AB - BACKGROUND: Alveolar epithelial cells are known as progenitor cells for the restoration from the damage in the lung. Calcitonin gene-related peptide (CGRP) has been reported to play an important role in the proliferation of various types of epithelial and endothelial cells. We investigated the effects of CGRP on the proliferation of alveolar epithelial cells in vitro and in vivo. METHODS: A549 cells were cultured in Dulbecco Modified Eagle Medium with 5% fatal bovin serum for 24 hours, then CGRP was added in vitro. The proliferation of DNA synthesis was measured using 5-bromo-2-deoxyuridine, an analog of thymidine, by enzyme linked immunosorbent assay.As one intracellular response to CGRP, we examined activation of p44/42- extracellular signal-regulated kinase (ERK) pathway by adding CGRP, using western blotting method.Recombinant adenovirus encoding nuclear-targeted-human beta-CGRP (rhCGRP) was administered into Male Wister rat (n = 5, 10 weeks old) lungs by intratracheal instillation in vivo. 7 days after the administration of CGRP, rat lungs were harvested and histological findings and immunohistochemical staining of proliferating cell nuclear antigen (PCNA) were evaluated to examine cell proliferation. RESULTS: In vitro study, CGRP increased the proliferation of A549 cells in a dose and time dependent manner. CGRP8-37 (inhibitor of CGRP receptor) decreased CGRP induced proliferation of DNA synthesis. Phosphorylation of ERK pathway was observed within 15 minutes and peaked in one hour. U0126 (inhibitor of ERK pathway) decreased CGRP induced proliferation of DNA synthesis.In vivo study, histological examination of the lung indicated proliferation of alveolar epithelial cells in the rhCGRP-treated group and the nuclei of alveolar epithelial cells were positive for PCNA immunostaining. CONCLUSION: In this study, we conclude that CGRP stimulates proliferation of human alveolar epithelial cells in vivo and in vitro. PMID- 19192278 TI - Specific gene expression profiles and chromosomal abnormalities are associated with infant disseminated neuroblastoma. AB - BACKGROUND: Neuroblastoma (NB) tumours have the highest incidence of spontaneous remission, especially among the stage 4s NB subgroup affecting infants. Clinical distinction of stage 4s from lethal stage 4 can be difficult, but critical for therapeutic decisions. The aim of this study was to investigate chromosomal alterations and differential gene expression amongst infant disseminated NB subgroups. METHODS: Thirty-five NB tumours from patients diagnosed at < 18 months (25 stage 4 and 10 stage 4s), were evaluated by allelic and gene expression analyses. RESULTS: All stage 4s patients underwent spontaneous remission, only 48% stage 4 patients survived despite combined modality therapy. Stage 4 tumours were 90% near-diploid/tetraploid, 44% MYCN amplified, 77% had 1p LOH (50% 1p36), 23% 11q and/or 14q LOH (27%) and 47% had 17q gain. Stage 4s were 90% near triploid, none MYCN amplified and LOH was restricted to 11q. Initial comparison analyses between stage 4s and 4 < 12 months tumours revealed distinct gene expression profiles. A significant portion of genes mapped to chromosome 1 (P < 0.0001), 90% with higher expression in stage 4s, and chromosome 11 (P = 0.0054), 91% with higher expression in stage 4. Less definite expression profiles were observed between stage 4s and 4 < 18m, yet, association with chromosomes 1 (P < 0.0001) and 11 (P = 0.005) was maintained. Distinct gene expression profiles but no significant association with specific chromosomal region localization was observed between stage 4s and stage 4 < 18 months without MYCN amplification. CONCLUSION: Specific chromosomal aberrations are associated with distinct gene expression profiles which characterize spontaneously regressing or aggressive infant NB, providing the biological basis for the distinct clinical behaviour. PMID- 19192279 TI - Is psychiatric emergency service (PES) use increasing over time? AB - BACKGROUND: Several recent studies have reported a significant increase in medical emergency department (ED) use for reasons of mental health. The diagnostic profile of these patients however differs from that usually described for patients visiting the psychiatric emergency service (PES). Few studies have specifically focused upon long-term PES utilization rates. Those that do typically present data from the early 80s, suggesting that deinstitutionalization may be an important contributing factor to the increases found. The aim of this study was to assess PES use using a more recent time frame and, the effects of non-specific factors, such as population growth, on this use. METHODS: Visits per year at several different types of PESs were obtained; (a) for an 11-year period at a general hospital PES while the surrounding population remained stable, (b) at that same PES while the catchment area population doubled over a period of a few years, (c) for an 11-year period at two PESs without catchment areas while the surrounding population increased and (d-) for a 12-year period at a PES in a mental health facility while the surrounding population increased. Moderately conservative criteria were used to define either a trend or, a significant increase in utilization rates. RESULTS: Each site had an inherent, 7 to 15% yearly variability in the number of PES visits. Over time however, only those where the surrounding population increased (either by an increase in the catchment area size or a regional increase in the population census) showed a trend or, a significant increase in utilization rates. These increases however were modest and of the order of 12 to 19%. CONCLUSION: Long observation periods are required in order to detect stable changes in PES utilization rates over time. As such, population growth may be but one of several factors underlying these increases. Organizational changes in mental health care delivery in the vicinity of the services that showed an increase could also have contributed. These latter would simply have redistributed (to the PES) the pre existing pool of mental health care patients, resulting in an increase that is more apparent than real. PMID- 19192280 TI - Is searching full text more effective than searching abstracts? AB - BACKGROUND: With the growing availability of full-text articles online, scientists and other consumers of the life sciences literature now have the ability to go beyond searching bibliographic records (title, abstract, metadata) to directly access full-text content. Motivated by this emerging trend, I posed the following question: is searching full text more effective than searching abstracts? This question is answered by comparing text retrieval algorithms on MEDLINE abstracts, full-text articles, and spans (paragraphs) within full-text articles using data from the TREC 2007 genomics track evaluation. Two retrieval models are examined: bm25 and the ranking algorithm implemented in the open source Lucene search engine. RESULTS: Experiments show that treating an entire article as an indexing unit does not consistently yield higher effectiveness compared to abstract-only search. However, retrieval based on spans, or paragraphs-sized segments of full-text articles, consistently outperforms abstract-only search. Results suggest that highest overall effectiveness may be achieved by combining evidence from spans and full articles. CONCLUSION: Users searching full text are more likely to find relevant articles than searching only abstracts. This finding affirms the value of full text collections for text retrieval and provides a starting point for future work in exploring algorithms that take advantage of rapidly-growing digital archives. Experimental results also highlight the need to develop distributed text retrieval algorithms, since full-text articles are significantly longer than abstracts and may require the computational resources of multiple machines in a cluster. The MapReduce programming model provides a convenient framework for organizing such computations. PMID- 19192281 TI - Evaluation of day one embryo quality and IVF outcome--a comparison of two scoring systems. AB - BACKGROUND: The aim of our retrospective study was to compare the clinical usefulness of two non-invasive embryo scoring systems based either on a simplified pronuclear morphology of the zygote or on early cleavage rate, as well as their combination, for the selection of embryos with the best implantation potential in embryo transfer (ET). METHODS: Over a period of five years, the quality of 2708 embryos from 364 IVF cycles in women under the age of 39 years was assessed using these scoring systems in a university assisted reproduction centre. ET was always performed on day 3 of cultivation. The outcome of ETs of 702 embryos scored in the respective systems or their combination was retrospectively analyzed in terms of biochemical (bPR) and clinical pregnancy rates (cPR) and implantation rate (IR). Mann-Whitney U test and t-test for differences between relative values were used, p < 0.05 was considered statistically significant. RESULTS: There was no difference in outcome parameters in 109 cycles where only Pattern "0" zygotes, according to our simplified pronuclear morphology classification, were transferred and 140 cycles where only "other" pattern zygotes were transferred, regardless of their cleavage rate. On the contrary, significantly greater cPR and IR (p = 0.003 and p = 0.006, respectively) were achieved in 120 cycles where only early cleavage (EC) embryos were transferred compared with 152 cycles where only non early cleavage (NEC) embryos were transferred regardless of their pronuclear morphology. The best outcome in terms of cPR (56%) and IR (43%) was found in 50 cycles when Pattern "0" and EC embryos only were used for transfer. CONCLUSION: The results indicate that early cleavage is a better independent marker of implantation potential than zygote morphology. The best outcome can be achieved if both embryo scoring systems are used jointly and the embryo is classified as EC and Pattern "0". PMID- 19192282 TI - A child presenting with acute renal failure secondary to a high dose of indomethacin: a case report. AB - INTRODUCTION: Acute renal failure caused by nonsteroidal anti-inflammatory drugs administered at therapeutic doses is generally mild, non-anuric and transitory. There are no publications on indomethacin toxicity secondary to high doses in children. The aim of this article is to describe acute renal failure secondary to a high dose of indomethacin in a child and to review an error in a supervised drug prescription and administration system. CASE PRESENTATION: Due to a medication error, a 20-day-old infant in the postoperative period of surgery for Fallot's tetralogy received a dose of 10 mg/kg of indomethacin, 50 to 100 times higher than the therapeutic dose. The child presented with acute, oligo-anuric renal failure requiring treatment with continuous venovenous renal replacement therapy, achieving complete recovery of renal function with no sequelae. CONCLUSION: In order to reduce medication errors in critically ill children, it is necessary to develop a supervised drug prescription and administration system, with controls at various levels. PMID- 19192283 TI - Perforation of Meckel's diverticulum caused by a chicken bone: a case report. AB - INTRODUCTION: Meckel's diverticulum represents a true diverticulum of the ileum containing all three layers of the bowel wall and is found on the wall of the distal ileum, usually about 2 feet from the ileocaecal valve. Although Meckel's diverticulum is a common congenital abnormality of the gastrointestinal tract, it is often difficult to diagnose. Patients with perforation of Meckel's diverticulum may present with right iliac fossa pain, which mimics acute appendicitis. CASE PRESENTATION: A 17-year-old man presented with a 3-day history of lower abdominal pain. On examination, the patient had tenderness in his right iliac fossa. A provisional diagnosis of acute appendicitis was made. The patient was taken to theatre for laparoscopy with the option of appendicectomy. The appendix was found to be normal. An inflamed and perforated Meckel's diverticulum was found to be the cause of the abdominal pain. Meckel's diverticulectomy was performed. The patient made an uneventful recovery and was discharged with further follow-up in the outpatient clinic. CONCLUSION: Complications of Meckel's diverticulum can be fatal and early recognition leads to appropriate management. This case report highlights the importance of considering Meckel's diverticulum as a differential diagnosis of acute abdomen in a young patient. PMID- 19192284 TI - Electrical wire as a foreign body in a male urethra: a case report. AB - INTRODUCTION: Self-inflicted foreign bodies in the male urethra and urinary bladder are an emergency that urologists may rarely have to face. A case of an electrical wire inserted in the male urethra and coiled in the bladder is presented. CASE PRESENTATION: A 53-year-old male presented with the inability to void and bloody urethral discharge after having introduced an electrical wire in his urethra for masturbation 3 hours earlier. He had made several unsuccessful attempts to remove it. CONCLUSION: The variety of these objects may be impressive and removal of the foreign body may be quite challenging requiring imagination and high-level surgical skills., In this case an electrical wire was used and the diagnostic as well as the therapeutic steps for its removal are presented. PMID- 19192285 TI - A general modular framework for gene set enrichment analysis. AB - BACKGROUND: Analysis of microarray and other high-throughput data on the basis of gene sets, rather than individual genes, is becoming more important in genomic studies. Correspondingly, a large number of statistical approaches for detecting gene set enrichment have been proposed, but both the interrelations and the relative performance of the various methods are still very much unclear. RESULTS: We conduct an extensive survey of statistical approaches for gene set analysis and identify a common modular structure underlying most published methods. Based on this finding we propose a general framework for detecting gene set enrichment. This framework provides a meta-theory of gene set analysis that not only helps to gain a better understanding of the relative merits of each embedded approach but also facilitates a principled comparison and offers insights into the relative interplay of the methods. CONCLUSION: We use this framework to conduct a computer simulation comparing 261 different variants of gene set enrichment procedures and to analyze two experimental data sets. Based on the results we offer recommendations for best practices regarding the choice of effective procedures for gene set enrichment analysis. PMID- 19192286 TI - Time to revisit Dr Finlay's casebook? The unique potential of the general practice case report. PMID- 19192287 TI - Comparative larval myogenesis and adult myoanatomy of the rhynchonelliform (articulate) brachiopods Argyrotheca cordata, A. cistellula, and Terebratalia transversa. AB - BACKGROUND: Despite significant methodological progress, Brachiopoda remains one of the lophotrochozoan phyla for which no recent ontogenetic data employing modern methodologies such as fluorescence labelling and confocal microscopy are available. This is particularly astonishing given the ongoing controversy concerning its phylogenetic position. In order to contribute new morphogenetic data for phylogenetic and evolutionary inferences, we describe herein the ontogeny and myoanatomy of larvae and adults of the rhynchonelliform brachiopods Argyrotheca cordata, A. cistellula, and Terebratalia transversa using fluorescence F-actin labelling combined with confocal laserscanning microscopy. RESULTS: Fully grown larvae of A. cordata and T. transversa consist of three distinct body regions, namely an apical lobe, a mantle lobe with four bundles of setae, and a pedicle lobe. Myogenesis is very similar in these two species. The first anlagen of the musculature develop in the pedicle lobe, followed by setae muscles and the mantle lobe musculature. Late-stage larvae show a network of strong pedicle muscles, central mantle muscles, longitudinal muscles running from the mantle to the pedicle lobe, setae pouch muscles, setae muscles, a U-shaped muscle, serial mantle muscles, and apical longitudinal as well as apical transversal muscles. Fully developed A. cistellula larvae differ from the former species in that they have only two visible body lobes and lack setae. Nevertheless, we found corresponding muscle systems to all muscles present in the former two species, except for the musculature associated with the setae, in larvae of A. cistellula. With our survey of the adult myoanatomy of A. cordata and A. cistellula and the juvenile muscular architecture of T. transversa we confirm the presence of adductors, diductors, dorsal and ventral pedicle adjustors, mantle margin muscles, a distinct musculature of the intestine, and striated muscle fibres in the tentacles for all three species. CONCLUSION: Our data indicate that larvae of rhynchonelliform brachiopods share a common muscular bodyplan and are thus derived from a common ancestral larval type. Comparison of the muscular phenotype of rhynchonelliform larvae to that of the other two lophophorate phyla, Phoronida and Ectoprocta, does not indicate homology of individual larval muscles. This may be due to an early evolutionary split of the ontogenetic pathways of Brachiopoda, Phoronida, and Ectoprocta that gave rise to the morphological diversity of these phyla. PMID- 19192288 TI - The role of menopause and reproductive senescence in a long-lived social mammal. AB - BACKGROUND: Menopause is a seemingly maladaptive life-history trait that is found in many long-lived mammals. There are two competing evolutionary hypotheses for this phenomenon; in the adaptive view of menopause, the cessation of reproduction may increase the fitness of older females; in the non-adaptive view, menopause may be explained by physiological deterioration with age. The decline and eventual cessation of reproduction has been documented in a number of mammalian species, however the evolutionary cause of this trait is unknown. RESULTS: We examined a unique 30-year time series of killer whales, tracking the reproductive performance of individuals through time. Killer whales are extremely long-lived, and may have the longest documented post-reproductive lifespan of any mammal, including humans. We found no strong support for either of the adaptive hypotheses of menopause; there was little support for the presence of post reproductive females benefitting their daughter's reproductive performance (interbirth interval and reproductive lifespan of daughters), or the number of mature recruits to the population. Oldest mothers (> 35) did appear to have a small positive impact on calf survival, suggesting that females may gain experience with age. There was mixed support for the grandmother hypothesis - grandoffspring survival probabilities were not influenced by living grandmothers, but grandmothers may positively influence survival of juveniles at a critical life stage. CONCLUSION: Although existing data do not allow us to examine evolutionary tradeoffs between survival and reproduction for this species, we were able to examine the effect of maternal age on offspring survival. Our results are consistent with similar studies of other mammals - oldest mothers appear to be better mothers, producing calves with higher survival rates. Studies of juvenile survival in humans have reported positive benefits of grandmothers on newly weaned infants; our results indicate that 3-year old killer whales may experience a positive benefit from helpful grandmothers. While our research provides little support for menopause evolving to provide fitness benefits to mothers or grandmothers, our work supports previous research showing that menopause and long post-reproductive lifespans are not a human phenomenon. PMID- 19192289 TI - Secretion of MCP-1 and other paracrine factors in a novel tumor-bone coculture model. AB - BACKGROUND: The bone-tumor microenvironment encompasses unique interactions between the normal cells of the bone and marrow cavity and the malignant cells from a primary or metastasized cancer. A multitude of paracrine factors within this microenvironment such as the growth factor, TGF-beta, and the chemokine, MCP 1, are secreted by many of these cell types. These factors can act in concert to modulate normal and malignant cell proliferation, malignant cell migration and invasion and, often, mediate bone cancer pain. Although many valuable in vitro and in vivo models exist, identifying the relevant paracrine factors and deciphering their interactions is still a challenge. The aim of our study is to test an ex vivo coculture model that will allow monitoring of the expression, release and regulation of paracrine factors during interactions of an intact femur explant and tumor cells. METHODS: Intact or marrow-depleted neonatal mouse femurs and select murine and human sarcoma or carcinoma cell lines were incubated singly or in coculture in specialized well plates. Viability of the bone and cells was determined by immunohistochemical stains, microscopy and marrow cytopreps. Secretion and mRNA expression of paracrine factors was quantitated by ELISA and real-time RT-PCR. RESULTS: Compartments of the bone were optimally viable for up to 48 h in culture and tumor cells for up to 4 days. Bone was the major contributor of TGF-beta and MMP2 whereas both bone and sarcoma cells secreted the chemokine MCP-1 in cocultures. Synergistic interaction between the femur and sarcoma resulted in enhanced MCP-1 secretion and expression in cocultures and was dependent on the presence of the hematopoietic component of the bone as well as other bone cells. In contrast, coculturing with breast carcinoma cells resulted in reduction of TGF-beta and MCP-1 secretion from the bone. CONCLUSION: These studies illustrate the feasibility of this model to examine paracrine interactions between intact bone and tumor cells. Further study of unique regulation of MCP-1 secretion and signaling between these cell types in different types of cancer will be possible using this simulated microenvironment. PMID- 19192290 TI - Subcutaneous emphysema, pneumomediastinum and pneumoperitoneum after unsuccessful ERCP: a case report. AB - BACKGROUND: The presence of subcutaneous emphysema, pneumomediastinum and pneumoperitoneum simultaneously is a rare complication of upper gastrointestinal endoscopy that usually indicates free perforation to the peritoneal cavity or the retroperitoneal space. CASE PRESENTATION: We report an unusual case of a self limited subcutaneous emphysema, pneumomediastinum and pneumoperitoneum following an unsuccessful ERCP for removal of a common bile duct stone. CONCLUSION: There was no radiological evidence of peritoneal or retroperitoneal perforation. This complication is distinct from pneumomediastinum and pneumoperitoneum due to perforation, and must be recognized, because it is benign and needs no surgical or radiological intervention. PMID- 19192291 TI - Comparative genomic analysis of light-regulated transcripts in the Solanaceae. AB - BACKGROUND: Plants use different light signals to adjust their growth and development to the prevailing environmental conditions. Studies in the model species Arabidopsis thaliana and rice indicate that these adjustments are mediated by large changes in the transcriptome. Here we compared transcriptional responses to light in different species of the Solanaceae to investigate common as well as species-specific changes in gene expression. RESULTS: cDNA microarrays were used to identify genes regulated by a transition from long days (LD) to short days (SD) in the leaves of potato and tobacco plants, and by phytochrome B (phyB), the photoreceptor that represses tuberization under LD in potato. We also compared transcriptional responses to photoperiod in Nicotiana tabacum Maryland Mammoth (MM), which flowers only under SD, with those of Nicotiana sylvestris, which flowers only under LD conditions. Finally, we identified genes regulated by red compared to far-red light treatments that promote germination in tomato. CONCLUSION: Most of the genes up-regulated in LD were associated with photosynthesis, the synthesis of protective pigments and the maintenance of redox homeostasis, probably contributing to the acclimatization to seasonal changes in irradiance. Some of the photoperiodically regulated genes were the same in potato and tobacco. Others were different but belonged to similar functional categories, suggesting that conserved as well as convergent evolutionary processes are responsible for physiological adjustments to seasonal changes in the Solanaceae. A beta-ZIP transcription factor whose expression correlated with the floral transition in Nicotiana species with contrasting photoperiodic responses was also regulated by photoperiod and phyB in potato, and is a candidate gene to act as a general regulator of photoperiodic responses. Finally, GIGANTEA, a gene that controls flowering time in Arabidopsis thaliana and rice, was regulated by photoperiod in the leaves of potato and tobacco and by red compared to far-light treatments that promote germination in tomato seeds, suggesting that a conserved light signaling cascade acts across developmental contexts and species. PMID- 19192292 TI - A split and rearranged nuclear gene encoding the iron-sulfur subunit of mitochondrial succinate dehydrogenase in Euglenozoa. AB - BACKGROUND: Analyses based on phylogenetic and ultrastructural data have suggested that euglenids (such as Euglena gracilis), trypanosomatids and diplonemids are members of a monophyletic lineage termed Euglenozoa. However, many uncertainties are associated with phylogenetic reconstructions for ancient and rapidly evolving groups; thus, rare genomic characters become increasingly important in reinforcing inferred phylogenetic relationships. FINDINGS: We discovered that the iron-sulfur subunit (SdhB) of mitochondrial succinate dehydrogenase is encoded by a split and rearranged nuclear gene in Euglena gracilis and trypanosomatids, an example of a rare genomic character. The two subgenic modules are transcribed independently and the resulting mRNAs appear to be independently translated, with the two protein products imported into mitochondria, based on the presence of predicted mitochondrial targeting peptides. Although the inferred protein sequences are in general very divergent from those of other organisms, all of the required iron-sulfur cluster coordinating residues are present. Moreover, the discontinuity in the euglenozoan SdhB sequence occurs between the two domains of a typical, covalently continuous SdhB, consistent with the inference that the euglenozoan 'half' proteins are functional. CONCLUSION: The discovery of this unique molecular marker provides evidence for the monophyly of Euglenozoa that is independent of evolutionary models. Our results pose questions about the origin and timing of this novel gene arrangement and the structure and function of euglenozoan SdhB. PMID- 19192294 TI - The commercial performance of cellulosic ethanol supply-chains in Europe. AB - BACKGROUND: The production of fuel-grade ethanol from lignocellulosic biomass resources has the potential to increase biofuel production capacity whilst minimising the negative environmental impacts. These benefits will only be realised if lignocellulosic ethanol production can compete on price with conventional fossil fuels and if it can be produced commercially at scale. This paper focuses on lignocellulosic ethanol production in Europe. The hypothesis is that the eventual cost of production will be determined not only by the performance of the conversion process but by the performance of the entire supply chain from feedstock production to consumption. To test this, a model for supply chain cost comparison is developed, the components of representative ethanol supply-chains are described, the factors that are most important in determining the cost and profitability of ethanol production are identified, and a detailed sensitivity analysis is conducted. RESULTS: The most important cost determinants are the cost of feedstocks, primarily determined by location and existing markets, and the value obtained for ethanol, primarily determined by the oil price and policy incentives. Both of these factors are highly uncertain. The best performing chains (ethanol produced from softwood and sold as a low percentage blend with gasoline) could ultimately be cost competitive with gasoline without requiring subsidy, but production from straw would generally be less competitive. CONCLUSION: Supply-chain design will play a critical role in determining commercial viability. The importance of feedstock supply highlights the need for location-specific assessments of feedstock availability and price. Similarly, the role of subsidies and policy incentives in creating and sustaining the ethanol market highlights the importance of political engagement and the need to include political risks in investment appraisal. For the supply-chains described here, and with the cost and market parameters selected, selling ethanol as a low percentage blend with gasoline will maximise ethanol revenues and minimise the need for subsidies. It follows, therefore, that the market for low percentage blends should be saturated before markets for high percentage blends. PMID- 19192293 TI - Emergence, development and diversification of the TGF-beta signalling pathway within the animal kingdom. AB - BACKGROUND: The question of how genomic processes, such as gene duplication, give rise to co-ordinated organismal properties, such as emergence of new body plans, organs and lifestyles, is of importance in developmental and evolutionary biology. Herein, we focus on the diversification of the transforming growth factor-beta (TGF-beta) pathway -- one of the fundamental and versatile metazoan signal transduction engines. RESULTS: After an investigation of 33 genomes, we show that the emergence of the TGF-beta pathway coincided with appearance of the first known animal species. The primordial pathway repertoire consisted of four Smads and four receptors, similar to those observed in the extant genome of the early diverging tablet animal (Trichoplax adhaerens). We subsequently retrace duplications in ancestral genomes on the lineage leading to humans, as well as lineage-specific duplications, such as those which gave rise to novel Smads and receptors in teleost fishes. We conclude that the diversification of the TGF-beta pathway can be parsimoniously explained according to the 2R model, with additional rounds of duplications in teleost fishes. Finally, we investigate duplications followed by accelerated evolution which gave rise to an atypical TGF beta pathway in free-living bacterial feeding nematodes of the genus Rhabditis. CONCLUSION: Our results challenge the view of well-conserved developmental pathways. The TGF-beta signal transduction engine has expanded through gene duplication, continually adopting new functions, as animals grew in anatomical complexity, colonized new environments, and developed an active immune system. PMID- 19192295 TI - Health status transitions in community-living elderly with complex care needs: a latent class approach. AB - BACKGROUND: For older persons with complex care needs, accounting for the variability and interdependency in how health dimensions manifest themselves is necessary to understand the dynamic of health status. Our objective is to test the hypothesis that a latent classification can capture this heterogeneity in a population of frail elderly persons living in the community. Based on a person centered approach, the classification corresponds to substantively meaningful groups of individuals who present with a comparable constellation of health problems. METHODS: Using data collected for the SIPA project, a system of integrated care for frail older people (n = 1164), we performed latent class analyses to identify homogenous categories of health status (i.e. health profiles) based on 17 indicators of prevalent health problems (chronic conditions; depression; cognition; functional and sensory limitations; instrumental, mobility and personal care disability) Then, we conducted latent transition analyses to study change in profile membership over 2 consecutive periods of 12 and 10 months, respectively. We modeled competing risks for mortality and lost to follow-up as absorbing states to avoid attrition biases. RESULTS: We identified four health profiles that distinguish the physical and cognitive dimensions of health and capture severity along the disability dimension. The profiles are stable over time and robust to mortality and lost to follow-up attrition. The differentiated and gender-specific patterns of transition probabilities demonstrate the profiles' sensitivity to change in health status and unmasked the differential relationship of physical and cognitive domains with progression in disability. CONCLUSION: Our approach may prove useful at organization and policy levels where many issues call for classification of individuals into pragmatically meaningful groups. In dealing with attrition biases, our analytical strategy could provide critical information for the planning of longitudinal studies of aging. Combined, these findings address a central challenge in geriatrics by making the multidimensional and dynamic nature of health computationally tractable. PMID- 19192296 TI - PeakSeeker: a program for interpreting genotypes of mononucleotide repeats. AB - BACKGROUND: Mononucleotide repeat microsatellites are abundant, highly polymorphic DNA sequences, having the potential to serve as valuable genetic markers. Use of mononucleotide microsatellites has been limited by their tendency to produce "stutter", confounding signals from insertions and deletions within the mononucleotide tract that occur during PCR, which complicates interpretation of genotypes by masking the true position of alleles. Consequently, microsatellites with larger repeating subunits (dinucleotide and trinucleotide motifs) are used, which produce less stutter but are less genetically heterogeneous and less informative. A method to interpret the genotypes of mononucleotide repeats would permit the widespread use of those highly informative microsatellites in genetic research. FINDINGS: We have developed an approach to interpret genotypes of mononucleotide repeats using a software program, named PeakSeeker. PeakSeeker interprets experimental electropherograms as the most likely product of signals from individual alleles. Because mononucleotide tracts demonstrate locus-specific patterns of stutter peaks, this approach requires that the genotype pattern from a single allele is defined for each marker, which can be approximated by genotyping single DNA molecules or homozygotes. We have evaluated the program's ability to discriminate various types of homozygous and heterozygous mononucleotide loci using simulated and experimental data. CONCLUSION: Mononucleotide tracts offer significant advantages over di- and tri-nucleotide microsatellite markers traditionally employed in genetic research. The PeakSeeker algorithm provides a high-throughput means to type mononucleotide tracts using conventional and widely implemented fragment length polymorphism genotyping. Furthermore, the PeakSeeker algorithm could potentially be adapted to improve, and perhaps to standardize, the analysis of conventional microsatellite genotypes. PMID- 19192297 TI - Characteristics of Epstein-Barr virus-associated gastric cancer: a study of 235 cases at a comprehensive cancer center in U.S.A. AB - BACKGROUND: Epstein-Barr virus (EBV) has been shown to be associated with gastric cancer. However, inconsistent findings have been reported regarding the distribution of EBV infected cells (in normal gastric epithelium vs. intestinal metaplastic cells vs. in neoplastic cells) and the characteristics of EBV associated gastric cancer. Lymph node positive EBV-associated gastric cancer has not been systematically studied. The aims of this study were to evaluate EBV associated gastric cancer, to assess the distribution of EBV infected cells including all positive lymph nodes, and to define the characteristics of EBV associated gastric cancer. DESIGN: The study included primary gastric cancer patients who underwent surgical resection with no preoperative treatment at M.D. Anderson Cancer Center between 1987 and 2006. Formalin-fixed paraffin-embedded tissue from these resection specimens were assessed for EBV by in situ hybridization, the gold standard for EBV detection in tissue. EBV status was analyzed along with clinicopathologic parameters including age, gender, tumor type, lymph node status, and pathologic stage of the tumor. RESULTS: Among 235 patients, 12 had intranuclear expression of EBV. EBV staining was seen only in tumor cells and no detectable EBV was observed in normal gastric mucosa, intestinal metaplasia or stromal cells. Eight of 12 patients with EBV-associated gastric cancer had regional lymph node metastasis. Of note, metastatic tumor cells in all of the involved lymph nodes of these 8 cases contained EBV. The epidemiologic data showed 11 of the 12 patients with EBV-associated gastric cancer were men, ranging in age from 54 to 78 years (mean age, 60 years; median age, 62.1 years). The age distribution for non-EBV associated gastric cancer patients ranged from 21 to 93 years (mean age, 67 years; median age, 66.4 years). CONCLUSION: Our study demonstrated that EBV is present exclusively in gastric cancer cells. The detection of EBV in tumor cells in all of the lymph nodes involved with metastatic gastric carcinoma suggests simultaneous replication of EBV and tumor cells. The predominantly male gender and relatively younger age observed for the EBV-infected gastric cancer cases suggest an association between this disease and other factors, such as life style. PMID- 19192298 TI - A comparative study of cultural methods for the detection of Salmonella in feed and feed ingredients. AB - BACKGROUND: Animal feed as a source of infection to food producing animals is much debated. In order to increase our present knowledge about possible feed transmission it is important to know that the present isolation methods for Salmonella are reliable also for feed materials. In a comparative study the ability of the standard method used for isolation of Salmonella in feed in the Nordic countries, the NMKL71 method (Nordic Committee on Food Analysis) was compared to the Modified Semisolid Rappaport Vassiliadis method (MSRV) and the international standard method (EN ISO 6579:2002). Five different feed materials were investigated, namely wheat grain, soybean meal, rape seed meal, palm kernel meal, pellets of pig feed and also scrapings from a feed mill elevator. Four different levels of the Salmonella serotypes S. Typhimurium, S. Cubana and S. Yoruba were added to each feed material, respectively. For all methods pre enrichment in Buffered Peptone Water (BPW) were carried out followed by enrichments in the different selective media and finally plating on selective agar media. RESULTS: The results obtained with all three methods showed no differences in detection levels, with an accuracy and sensitivity of 65% and 56%, respectively. However, Muller-Kauffmann tetrathionate-novobiocin broth (MKTTn), performed less well due to many false-negative results on Brilliant Green agar (BGA) plates. Compared to other feed materials palm kernel meal showed a higher detection level with all serotypes and methods tested. CONCLUSION: The results of this study showed that the accuracy, sensitivity and specificity of the investigated cultural methods were equivalent. However, the detection levels for different feed and feed ingredients varied considerably. PMID- 19192299 TI - GOrilla: a tool for discovery and visualization of enriched GO terms in ranked gene lists. AB - BACKGROUND: Since the inception of the GO annotation project, a variety of tools have been developed that support exploring and searching the GO database. In particular, a variety of tools that perform GO enrichment analysis are currently available. Most of these tools require as input a target set of genes and a background set and seek enrichment in the target set compared to the background set. A few tools also exist that support analyzing ranked lists. The latter typically rely on simulations or on union-bound correction for assigning statistical significance to the results. RESULTS: GOrilla is a web-based application that identifies enriched GO terms in ranked lists of genes, without requiring the user to provide explicit target and background sets. This is particularly useful in many typical cases where genomic data may be naturally represented as a ranked list of genes (e.g. by level of expression or of differential expression). GOrilla employs a flexible threshold statistical approach to discover GO terms that are significantly enriched at the top of a ranked gene list. Building on a complete theoretical characterization of the underlying distribution, called mHG, GOrilla computes an exact p-value for the observed enrichment, taking threshold multiple testing into account without the need for simulations. This enables rigorous statistical analysis of thousand of genes and thousands of GO terms in order of seconds. The output of the enrichment analysis is visualized as a hierarchical structure, providing a clear view of the relations between enriched GO terms. CONCLUSION: GOrilla is an efficient GO analysis tool with unique features that make a useful addition to the existing repertoire of GO enrichment tools. GOrilla's unique features and advantages over other threshold free enrichment tools include rigorous statistics, fast running time and an effective graphical representation. GOrilla is publicly available at: http://cbl-gorilla.cs.technion.ac.il PMID- 19192300 TI - Nitrous oxide may not increase the risk of cancer recurrence after colorectal surgery: a follow-up of a randomized controlled trial. AB - BACKGROUND: Even the best cancer surgery is usually associated with minimal residual disease. Whether these remaining malignant cells develop into clinical recurrence is at least partially determined by adequacy of host defense, especially natural killer cell function. Anesthetics impair immune defenses to varying degrees, but nitrous oxide appears to be especially problematic. We therefore tested the hypothesis that colorectal-cancer recurrence risk is augmented by nitrous oxide administration during colorectal surgery. METHODS: We conducted a 4- to 8-year follow-up of 204 patients with colorectal cancer who were randomly assigned to 65% nitrous oxide (n = 97) or nitrogen (n = 107), balanced with isoflurane and remifentanil. The primary outcome was the time to cancer recurrence. Our primary analysis was a multivariable Cox-proportional hazards regression model that included relevant baseline variables. In addition to treatment group, the model considered patient age, tumor grade, dissemination, adjacent organ invasion, vessel invasion, and the number of nodes involved. The study had 80% power to detect a 56% or greater reduction in recurrence rates (i.e., hazard ratio of 0.44 or less) at the 0.05 significance level. RESULTS: After adjusting for significant baseline covariables, risk of recurrence did not differ significantly for nitrous oxide and nitrogen, with a hazard ratio estimate (95% CI) of 1.10 (0.66, 1.83), P = 0.72. No two-way interactions with the treatment were statistically significant. CONCLUSION: Colorectal-cancer recurrence risks were not greatly different in patients who were randomly assigned to 65% nitrous oxide or nitrogen during surgery. Our results may not support avoiding nitrous oxide use to prevent recurrence of colorectal cancer. IMPLICATIONS STATEMENT: The risk of colorectal cancer recurrence was similar in patients who were randomly assigned to 65% nitrous oxide or nitrogen during colorectal surgery. TRIAL REGISTRATION: Current Controlled Clinical Trials NCT00781352 http://www.clinicaltrials.gov. PMID- 19192302 TI - The taxonomic status of the endangered thin-spined porcupine, Chaetomys subspinosus (Olfers, 1818), based on molecular and karyologic data. AB - BACKGROUND: The thin-spined porcupine, also known as the bristle-spined rat, Chaetomys subspinosus (Olfers, 1818), the only member of its genus, figures among Brazilian endangered species. In addition to being threatened, it is poorly known, and even its taxonomic status at the family level has long been controversial. The genus Chaetomys was originally regarded as a porcupine in the family Erethizontidae, but some authors classified it as a spiny-rat in the family Echimyidae. Although the dispute seems to be settled in favor of the erethizontid advocates, further discussion of its affinities should be based on a phylogenetic framework. In the present study, we used nucleotide-sequence data from the complete mitochondrial cytochrome b gene and karyotypic information to address this issue. Our molecular analyses included one individual of Chaetomys subspinosus from the state of Bahia in northeastern Brazil, and other hystricognaths. RESULTS: All topologies recovered in our molecular phylogenetic analyses strongly supported Chaetomys subspinosus as a sister clade of the erethizontids. Cytogenetically, Chaetomys subspinosus showed 2n = 52 and FN = 76. Although the sexual pair could not be identified, we assumed that the X chromosome is biarmed. The karyotype included 13 large to medium metacentric and submetacentric chromosome pairs, one small subtelocentric pair, and 12 small acrocentric pairs. The subtelocentric pair 14 had a terminal secondary constriction in the short arm, corresponding to the nucleolar organizer region (Ag-NOR), similar to the erethizontid Sphiggurus villosus, 2n = 42 and FN = 76, and different from the echimyids, in which the secondary constriction is interstitial. CONCLUSION: Both molecular phylogenies and karyotypical evidence indicated that Chaetomys is closely related to the Erethizontidae rather than to the Echimyidae, although in a basal position relative to the rest of the Erethizontidae. The high levels of molecular and morphological divergence suggest that Chaetomys belongs to an early radiation of the Erethizontidae that may have occurred in the Early Miocene, and should be assigned to its own subfamily, the Chaetomyinae. PMID- 19192303 TI - Plasticity of NMDA receptor NR2B subunit in memory and chronic pain. AB - Glutamatergic synapses play critical roles in brain functions and diseases. Long term potentiation (LTP) is a most effective cellular model for investigating the synaptic changes that underlie learning as well as brain disease--although different molecular mechanisms are likely involved in LTP in physiological and pathological conditions. In the case of learning, N-methyl-D-aspartate (NMDA) receptor is known to be important for triggering learning-related plasticity; alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic (AMPA) receptors are thought to be important for the expression of synaptic changes. In this review, I will examine recent evidence on the novel roles of NMDA receptors, in particular NR2B subunit-containing NMDA receptors in learning and chronic pain. A positive feedback control of NR2B receptor subunit is proposed to explain cortical sensitization involved in chronic pain, but not learning and memory. PMID- 19192304 TI - Threat of an influenza pandemic: family physicians in the front line. AB - BACKGROUND: The chance of an influenza pandemic is real and clinicians should keep themselves informed about the rationale and science behind preventive and therapeutic principles relating to an (impending) influenza pandemic. DISCUSSION: Vaccination is considered the best prevention in case of a pandemic threat and first choice to contain the impact of a pandemic. Pending the availability of an effective pandemic vaccine, antivirals are likely the only effective agents for prevention and treatment. When an influenza pandemic is impending, all interventions aim to prevent people becoming infected and to suppress replication and transmission of the virus as much as possible. Antivirals will be prescribed to patients with laboratory confirmed pre-pandemic influenza as well as to their contacts (post-exposure prophylaxis) which may delay development of or even prevent a pandemic. During a manifest influenza pandemic, however, there is large scale spreading of the influenza virus. Therefore, preventive use of antivirals is less efficient to prevent transmission. Delaying the pandemic is then important in order to prevent exhausting public health resources and disruption of society. Thus, during a manifest pandemic everyone with influenza symptoms should receive antivirals as quickly as possible, regardless of virological confirmation. To ensure optimal effectiveness of antivirals and to minimize development of drug resistant viral strains, the use of antivirals for annual influenza should be restrictive. The crucial position of family physicians during an (impending) influenza pandemic necessitates the development of primary health care guidelines on this topic for all countries. SUMMARY: Family physicians will play a key role in assessing and treating victims of a new influenza virus, and in reassuring the worried well. We outline various possible interventions in the event of an impending and a manifest influenza pandemic, such as non-medial measures, prescription of antivirals, and vaccination, and emphasize the need for pandemic influenza preparedness. PMID- 19192305 TI - Characterization of rag1 mutant zebrafish leukocytes. AB - BACKGROUND: Zebrafish may prove to be one of the best vertebrate models for innate immunology. These fish have sophisticated immune components, yet rely heavily on innate immune mechanisms. Thus, the development and characterization of mutant and/or knock out zebrafish are critical to help define immune cell and immune gene functions in the zebrafish model. The use of Severe Combined Immunodeficient (SCID) and recombination activation gene 1 and 2 mutant mice has allowed the investigation of the specific contribution of innate defenses in many infectious diseases. Similar zebrafish mutants are now being used in biomedical and fish immunology related research. This report describes the leukocyte populations in a unique model, recombination activation gene 1-/- mutant zebrafish (rag1 mutants). RESULTS: Differential counts of peripheral blood leukocytes (PBL) showed that rag1 mutants had significantly decreased lymphocyte like cell populations (34.7%) compared to wild-types (70.5%), and significantly increased granulocyte populations (52.7%) compared to wild-types (17.6%). Monocyte/macrophage populations were similar between mutants and wild-types, 12.6% and 11.3%, respectively. Differential leukocyte counts of rag1 mutant kidney hematopoietic tissue showed a significantly reduced lymphocyte-like cell population (8%), a significantly increased myelomonocyte population (57%), 34.8% precursor cells, and 0.2% thrombocytes, while wild-type hematopoietic kidney tissue showed 29.4% lymphocytes/lymphocyte-like cells, 36.4% myelomonocytes, 33.8% precursors and 0.5% thrombocytes. Flow cytometric analyses of kidney hematopoietic tissue revealed three leukocyte populations. Population A was monocytes and granulocytes and comprised 34.7% of the gated cells in rag1 mutants and 17.6% in wild-types. Population B consisted of hematopoietic precursors, and comprised 50% of the gated cells for rag1 mutants and 53% for wild-types. Population C consisted of lymphocytes and lymphocyte-like cells and comprised 7% of the gated cells in the rag1 mutants and 26% in the wild-types. Reverse transcriptase polymerase chain reaction (RT-PCR) assays demonstrated rag1 mutant kidney hematopoietic tissue expressed mRNA encoding Non-specific Cytotoxic cell receptor protein-1 (NCCRP-1) and Natural Killer (NK) cell lysin but lacked T cell receptor (TCR) and immunoglobulin (Ig) transcript expression, while wild-type kidney hematopoietic tissue expressed NCCRP-1, NK lysin, TCR and Ig transcript expression. CONCLUSION: Our study demonstrates that in comparison to wild-type zebrafish, rag1 mutants have a significantly reduced lymphocyte-like cell population that likely includes Non-specific cytotoxic cells (NCC) and NK cells (and lacks functional T and B lymphocytes), a similar macrophage/monocyte population, and a significantly increased neutrophil population. These zebrafish have comparable leukocyte populations to SCID and rag 1 and/or 2 mutant mice, that possess macrophages, natural killer cells and neutrophils, but lack T and B lymphocytes. Rag1 mutant zebrafish will provide the platform for remarkable investigations in fish and innate immunology, as rag 1 and 2 mutant mice did for mammalian immunology. PMID- 19192306 TI - Pseudomonas aeruginosa vesicles associate with and are internalized by human lung epithelial cells. AB - BACKGROUND: Pseudomonas aeruginosa is the major pathogen associated with chronic and ultimately fatal lung infections in patients with cystic fibrosis (CF). To investigate how P. aeruginosa-derived vesicles may contribute to lung disease, we explored their ability to associate with human lung cells. RESULTS: Purified vesicles associated with lung cells and were internalized in a time- and dose dependent manner. Vesicles from a CF isolate exhibited a 3- to 4-fold greater association with lung cells than vesicles from the lab strain PAO1. Vesicle internalization was temperature-dependent and was inhibited by hypertonic sucrose and cyclodextrins. Surface-bound vesicles rarely colocalized with clathrin. Internalized vesicles colocalized with the endoplasmic reticulum (ER) marker, TRAPalpha, as well as with ER-localized pools of cholera toxin and transferrin. CF isolates of P. aeruginosa abundantly secrete PaAP (PA2939), an aminopeptidase that associates with the surface of vesicles. Vesicles from a PaAP knockout strain exhibited a 40% decrease in cell association. Likewise, vesicles from PAO1 overexpressing PaAP displayed a significant increase in cell association. CONCLUSION: These data reveal that PaAP promotes the association of vesicles with lung cells. Taken together, these results suggest that P. aeruginosa vesicles can interact with and be internalized by lung epithelial cells and contribute to the inflammatory response during infection. PMID- 19192307 TI - Measuring the health impact of human rights violations related to Australian asylum policies and practices: a mixed methods study. AB - BACKGROUND: Human rights violations have adverse consequences for health. However, to date, there remains little empirical evidence documenting this association, beyond the obvious physical and psychological effects of torture. The primary aim of this study was to investigate whether Australian asylum policies and practices, which arguably violate human rights, are associated with adverse health outcomes. METHODS: We designed a mixed methods study to address the study aim. A cross-sectional survey was conducted with 71 Iraqi Temporary Protection Visa (TPV) refugees and 60 Iraqi Permanent Humanitarian Visa (PHV) refugees, residing in Melbourne, Australia. Prior to a recent policy amendment, TPV refugees were only given temporary residency status and had restricted access to a range of government funded benefits and services that permanent refugees are automatically entitled to. The quantitative results were triangulated with semi structured interviews with TPV refugees and service providers. The main outcome measures were self-reported physical and psychological health. Standardised self report instruments, validated in an Arabic population, were used to measure health and wellbeing outcomes. RESULTS: Forty-six percent of TPV refugees compared with 25% of PHV refugees reported symptoms consistent with a diagnosis of clinical depression (p = 0.003). After controlling for the effects of age, gender and marital status, TPV status made a statistically significant contribution to psychological distress (B = 0.5, 95% CI 0.3 to 0.71, p